TREA

3-ARYL-5-SUBSTITUTED-ISOQUINOLIN-1-ONE COMPOUNDS AND THEIR THERAPEUTIC USE

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Information

  • Patent Application
  • 20150099732
  • Publication Number
    20150099732
  • Date Filed
    March 07, 2013
    11 years ago
  • Date Published
    April 09, 2015
    9 years ago
Information
Abstract
The present invention pertains generally to the field of therapeutic compounds. More specifically the present invention pertains to certain 3-aryl-5-substituted-2H-isoquinolin-1-one compounds that, inter alia, inhibit PARP (e.g., PARP1, TNKS1, TNKS2, etc.) and/or Wnt signalling. The present invention also pertains to pharmaceutical compositions comprising such compounds, and the use of such compounds and compositions, both in vitro and in vivo, to inhibit PARP (e.g., PARP1, TNKS1, TNKS2, etc.); to inhibit Wnt signalling; to treat disorders that are ameliorated by the inhibition of PARP (e.g., PARP1, TNKS1, TNKS2, etc.); to treat disorders that are ameliorated by the inhibition of Wnt signalling; to treat proliferative conditions such as cancer, etc.
Description
TECHNICAL FIELD

The present invention pertains generally to the field of therapeutic compounds. More specifically the present invention pertains to certain 3-aryl-5-substituted-2H-isoquinolin-1-one compounds that, inter alia, inhibit PARP (e.g., PARP1, TNKS1, TNKS2, etc.) and/or Wnt signalling. The present invention also pertains to pharmaceutical compositions comprising such compounds, and the use of such compounds and compositions, both in vitro and in vivo, to inhibit PARP (e.g., PARP1, TNKS1, TNKS2, etc.); to inhibit Wnt signalling; to treat disorders that are ameliorated by the inhibition of PARP (e.g., PARP1, TNKS1, TNKS2, etc.); to treat disorders that are ameliorated by the inhibition of Wnt signalling; to treat proliferative conditions such as cancer, etc.


BACKGROUND

A number of publications are cited herein in order to more fully describe and disclose the invention and the state of the art to which the invention pertains. Each of these references is incorporated herein by reference in its entirety into the present disclosure, to the same extent as if each individual reference was specifically and individually indicated to be incorporated by reference.


Throughout this specification, including the claims which follow, unless the context requires otherwise, the word “comprise,” and variations such as “comprises” and “comprising,” will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.


It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a pharmaceutical carrier” includes mixtures of two or more such carriers, and the like.


Ranges are often expressed herein as from “about” one particular value, and/or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by the use of the antecedent “about,” it will be understood that the particular value forms another embodiment.


This disclosure includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.


Cancer

Cancer is the second largest cause of death worldwide. Cancer accounts for 13% of global mortality with more than 70% of cancer deaths occurring in low and middle-income countries where the prevalence of cancer is expected to increase as mortality from other diseases decreases. In the UK alone, a disease such as breast cancer kills over 12,000 women each year.


One approach to this problem has been to identify novel targets for cancer therapies and to use these to tailor the treatment of each patient according to the molecular make-up of their particular disease, rather than their overt clinical characteristics. While this has been in part successful, there are still a significant number of tumour types for which there are no targeted therapies and few treatment options other than surgery and cytotoxic chemotherapy.


PARP

There is now a significant body of evidence to suggest that inhibition of poly ADP ribose polymerase (PARP) superfamily proteins, such as PARP1, PARP2, Tankyrase 1 (also known as TNKS1, PARP5a) and Tankyrase 2 (also known as TNKS2, PARP5B) could have clinical utility. See, e.g., Krishnakumar et al., 2010. PARP superfamily members use beta-NAD+ as a substrate to generate ADP-ribose polymers on amino acid residues of protein acceptors. The result is a dramatic post-translational modification that can significantly alter the properties of the protein acceptor. See, e.g., Krishnakumar et al., 2010.


Although much of the focus has been on PARP1, studies over the past decade have identified a family of as many as 17 proteins that share homology to the catalytic domain of PARP1. In addition to the PARP-like domain, the PARP family members are “functionalized” with a wide variety of other structural and functional domains (e.g., DBDs, RNA-binding domains, subcellular localization signals, macrodomains, BRCT motifs, ankyrin repeats, zinc fingers) that determine their overall biological activities. Recently, a unified nomenclature referring to this family of proteins as ADP-ribosyl transferases (ARTs) has been proposed to recognize that fact that (1) PARPs catalyze a transferase reaction, not a template-dependent polymerization reaction; and (2) not all family members have PARP activity; some are likely to function as mono(ADP-ribosyl) transferases (mARTs). This new nomenclature is reflected in a recent structure-based classification of PARP family members into three groups based on their catalytic domains: (1) PARPs 1-5, which are bona fide PARPs containing a conserved glutamate (Glu 988 in PARP1) that defines the PARP catalytic activity; (2) PARPs 6-8, 10-12, and 14-16, which are confirmed or putative mARTs; and (3) PARPs 9 and 13, which lack key NAD-binding residues and the catalytic glutamate, and are likely inactive. See, e.g., Krishnakumar et al., 2010.


PARP family members localize to various cellular compartments, including the nucleus, cytoplasm, mitochondria, and vault particles, although the subcellular localization and function of many of the PARPs are unknown. The known functions of the PARP family members span a wide range of cellular processes, including DNA repair, transcription, cellular signalling, cell-cycle regulation, and mitosis. This diverse array of processes plays key roles in a wide variety of biological outcomes, including differentiation, development, stress responses, inflammation, and cancer. See, e.g., Krishnakumar et al., 2010.


The primary nuclear PARPs are PARP1, PARP2 (the closest paralog to PARP1), PARP3, and tankyrases 1 and 2. PARP1 is a very well studied protein and has a well-established role in DNA repair. See, e.g., Lord et al., 2008. Tankyrase 1 encompasses four distinct domains; the N terminal HPS domain (homopolymeric stretches of His, Pro and Ser); the ankyrin domain, containing 24 ANK repeats; a SAM (sterile alpha module) domain; and a C terminal PARP catalytic domain. See, e.g., Hsiao et al., 2008.


The best characterised function of tankyrase 1 is in telomere maintenance. The cellular machinery that normally replicates genomic DNA is unable to synthesise DNA at the telomere, the structure that caps the end of each chromosome. DNA synthesis at the telomere is instead carried out by telomerase. This enzyme complex consists of a RNA template and a DNA polymerase catalytic subunit. However, the activity of telomerase in most human somatic cells is relatively low and as such, attrition of the DNA at the telomere gradually occurs. This attrition of telomeric DNA is one of the factors that can lead to replicative senescence in somatic cells and this shortening of telomeres is often referred to as a “mitotic clock” that predetermines the replicative capacity of most cells. However, the situation in cancer cells is considerably different from that in somatic cells; up to 90% of all human cancer cells have a high level of telomerase activity. This increased level of telomere maintenance is one of the factors that enables tumour cells to avoid senescence and perpetually replicate. See, e.g., Harley, 2008.


The length of telomeric DNA is determined by a “protein counting” mechanism in which a series of telomere-bound proteins negatively regulate the access of telomerase to the telomere. For example, longer telomeres bind a larger number of DNA double strand-binding Telomeric Repeat Binding Factor (TRF1) proteins. Together with the TIN2-TPP1-POT1 protein complex, TRF1 blocks the access of telomerase to the 3′ DNA overhang at the end of chromosomes, thus limiting further extension of the telomere. Regulation of this process is controlled by tankyrase 1 which promotes telomeric extension by poly(ADP-ribosyl)ating TRF1, causing its release from the telomere and eventual proteasomal destruction. This release and degradation of TRF1 allows an enhanced level of telomerase access to the chromosome end and extension of the telomere. See, e.g., Harley, 2008.


Tankyrase 1 is also required after DNA replication in the S/G2 phase of the cell cycle to resolve sister chromatid cohesion before mitosis ensues. Depletion of tankyrase 1 in HeLa cells results in mitotic arrest. Persistent sister chromatid cohesion in tankyrase 1 depleted cells results in sister chromatid fusion. See, e.g., Hsiao et al., 2009. The mitotic defect in tankyrase-depleted cells may, in part, be determined by the tankyrase 1-mediated poly(ADP ribosyl)ation of the protein NuMA, which plays an essential role in organising microtubules at spindle pores. See, e.g., Chang et al., 2005.


Recent work has also suggested a role for Tankyrase 1 in the control of oncogenic Wnt signalling, most likely via a mechanism that involves the stabilisation of the Wnt signalling component, Axin. See, e.g., Huang et al., 2009. In this latter work and subsequent work (see, e.g., James et al., 2012; Bao et al., 2012; Casás-Selves et al., 2012; Waaler et al., 2012; Riffell et al., 2012) a number of investigators have shown that toolbox, non-drug like small molecule inhibitors of tankyrase can inhibit oncogenic Wnt signalling and can inhibit tumour cells that are addicted to Wnt signalling.


Wnt Signalling

Wnt signalling is an intracellular protein signalling network that transduces signals from cell surface bound receptors to a series of gene transcription events. In canonical Wnt signalling, Wnt ligands bind to cell-surface receptors of the Frizzled family; Frizzled bound receptors activate Dishevelled family proteins. In turn, activated Dishevelled proteins inhibit the function of a complex of proteins including Axin 1 and 2, GSK-3, and the protein APC. This Axin/GSK-3/APC complex normally promotes the proteolytic degradation of the β-catenin intracellular signalling molecule. When Wnt signalling is stimulated and Dishevelled proteins are active, the “β-catenin destruction complex” is inhibited, β-catenin degradation is reduced and β-catenin is able to enter the nucleus and interact with TCF/LEF family transcription factors. This latter act drives a series of specific gene expression events that ultimately mediate Wnt signalling.


The association of dysregulated Wnt/β-catenin signalling with cancer has been well documented. Constitutively activated β-catenin signalling, caused either by APC deficiency or activating β-catenin mutations can lead to tumourigenesis. Furthermore, tankyrase is directly involved in the Wnt signalling cascade. Tankyrase PARylates both Axin 1 and Axin 2 and causes their degradation, driving β-catenin stabilisation/nuclear translocation and TCF/LEF mediated transcription. See, e.g., Huang et al., 2009. When tankyrase is inhibited, either genetically or with small molecules, Axin1 and 2 levels are stabilized and β-catenin degradation is enhanced, ultimately suppressing Wnt signalling, even in situations where Wnt signalling is usually constitutively elevated, such as APC deficiency. See, e.g., Huang et al., 2009. These data suggest that tankyrase inhibition could be used in order to modulate Wnt signalling, both in cancer, but also in other, non-cancer, pathologies where Wnt signalling is aberrant.


In addition to its effects on Wnt signally, it has also recently been demonstrated that silencing of tankyrase 1 by RNA interference is lethal in tumour cells with deficiencies in either of the breast cancer susceptibility proteins, BRCA1 and BRCA2, but not in wild type cells. BRCA mutation carriers with cancer still retain functional BRCA protein function in their normal cells, whilst it is lacking in tumour cells, suggesting that a tankyrase 1 inhibitor could be used to selectively target tumour cells in BRCA patients. See, e.g., McCabe et al., 2009b. This approach of combining tumour-specific genetic deficiencies with inhibition of a drug target to elicit a therapeutic window is an example of a “synthetic lethal” approach to the design of cancer therapies. See, e.g., Kaelin, 2009. This BRCA selective effect of tankyrase 1 inhibition may be caused by telomere attrition (caused by tankyrase 1 inhibition) and stalled replication forks (caused by BRCA deficiency) acting in concert to cause a threshold of DNA damage that is inconsistent with cell viability. Alternatively, synergistic defects in cytokinesis and sister chromatid segregation caused by BRCA deficiency and tankyrase 1 inhibition may also underlie the BRCA selective effect. See, e.g., Daniels, 2004. The use of tankyrase 1 inhibition in this context is described in McCabe et al., 2009a and McCabe et al., 2009b.


It has been shown that a proportion of patients without BRCA mutations have clinical characteristics, tumour morphologies and tumour molecular profiles that are reminiscent of BRCA mutation-associated cancer, a property termed BRCAness. See, e.g., Turner et al., 2004. This BRCAness phenotype is most well described in a significant number of patients with triple negative breast tumours. See, e.g., Turner et al., 2004. It has been shown that BRCA1 deficient, triple-negative breast cancer cell lines such as HCC1937 are particularly sensitive to tankyrase 1 inhibition. See, e.g., McCabe et al., 2009a and McCabe et al., 2009b. Inhibiting tankyrase 1 therefore, may be very effective in patients with germ-line BRCA mutations as well as patients whose tumours exhibit a BRCAness phenotype.


Non-Tumourigenic Mechanisms Modulated by Tankyrase

In addition to tankyrase inhibitors having potential as cancer therapeutics, a number of other studies suggest tankyrase inhibitors could be used in a number of other non-cancer related pathologies, the majority of which are driven by aberrant Wnt signalling, of which tankyrase activity is a rate limiting step (see, e.g., Riffell et al., 2012).


For example:


Recent work has indicated that inhibition of tankyrase can stabilize Axin2 levels in immature oligodendrocyte progenitor cells (OLPs) (see, e.g., Fancy et al., 2011). On the basis that Axin2 function is essential for normal kinetics of remyelination, tankyrase inhibition has been shown to accelerate OLP myelination after hypoxic and demyelinating injury (see, e.g., Fancy et al., 2011). This data suggest that small molecule tankyrase inhibitors might serve as pharmacological agents that could aid remyelination in neuropathies such as multiple sclerosis, neonatal hypoxic ischemic encephalopathy (HIE), and neonatal periventricular leukomalacia (PVL) (see, e.g., Fancy et al., 2011).


Other studies have also shown that tankyrase is essential for Herpes Simplex Virus replication (HSV). Efficient HSV-1 replication requires tankyrase PARP activity (see, e.g., Li et al., 2011). Further support for this hypothesis comes from the observation that HSV did not replicate efficiently in cells depleted of tankyrase 1. Moreover, tankyrase and the tankyrase substrate TRF2 (telomeric repeat binding factor 2) control the degradation of Ebstein-Barr Virus (EBV) DNA (see, e.g., Deng et al., 2002), suggesting tankyrase inhibitors could have utility as antiviral agents.


In addition, tankyrase inhibition is known to modulate glucose uptake (see, e.g., Yeh et al., 2007), suggesting that a small molecule tankyrase inhibitor could have utility in the treatment of metabolic diseases such as type 2 diabetes. In this case, tankyrase inhibition is thought to modulate glucose uptake by altering the function and cellular localisation of the glucose transporter type 4 (GLUT4) and the aminopeptidase IRAP (insulin-responsive aminopeptidase).


In addition, tankyrase inhibition is known to induce cardiomyocyte differentiation (see, e.g., Wang et al., 2011), suggesting that small molecule tankyrase inhibitors could have some ability in the treatment of cardiac disorders, such as cardiac repair after cardiac infarction.


In addition, tankyrase inhibition is know to minimise the pathological effects of lung fibrosis and tankyrase inhibitors can improve the survival of mice with bleomycin induced lung fibrosis (see, e.g., Distler et al., 2012) suggesting that small molecule tankyrase inhibitors could have some usefuleness in the treatment of lung disorders and fibrotic disorders such as pulmonary fibrosis, cystic fibrosis, cirrhosis, endomyocardial fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, Crohn's disease, keloid, scleroderma/systemic sclerosis and arthrofibrosis.


In addition to these pathologies, Wnt signalling and its modulation are also involved in a number of other pathogenic conditions suggesting that small molecules tankyrase inhibitors could have utility in these other Wnt related diseases, including:

    • Alzheimer's disease, where the Wnt mediator B-catenin activity is aberrant (see, e.g., Caricasole et al., 2003; Moon et al., 2004; Mudher and Lovestone, 2002);
    • Dupuytren skin disease, where the Wnt mediator B-catenin activity is also aberrant (see, e.g., Varallo et al., 2003);
    • tooth agenesis, where the Wnt mediator Axin2 activity is aberrant (see, e.g., Lammi et al., 2004);
    • osteoarthritis, where the Wnt mediator secreted frizzled-related protein 3 (FRP3) activity is aberrant (see, e.g., Loughlin et al., 2004);
    • exudative vitreoretinopathy, where the Wnt mediators frizzled family receptor 4 (FZD4) (see, e.g., Robitaille et al., 2002) and Norrie disease protein (see, e.g., Xu et al., 2004) activities are aberrant;
    • schizophrenia, where the Wnt mediators glycogen synthase kinase 3 beta (GSK3b) and wingless-type MMTV integration site family member 1 (Wnt1) are aberrant (see, e.g., Kozlovsky et al., 2002; Miyaoka et al., 1999);
    • osteoporosis, where the Wnt mediator low density lipoprotein receptor-related protein 5 (LRP5) activity is aberrant (see, e.g., Gong et al., 2001);
    • dermal hypoplasia, where the Wnt mediator porcupine homolog (PORCN) activity is aberrant (see, e.g., Grzeschik et al., 2007);
    • XX sex reversal, where the Wnt mediator R-spondin 1 (RSPO1) activity is aberrant (see, e.g., Parma et al., 2006);
    • anonychia and hyponychia, were the Wnt mediator R-spondin 4 (RSPO4) is aberrant (see, e.g., Bergmann et al., 2006; Blaydon et al., 2006);
    • sclerosteosis and Van Buchem disease, where the Wnt mediator sclerostin (SOST) activity is aberrant (see, e.g., Balemans et al., 2001; Balemans et al., 2002);
    • Fuhrmann syndrome, were the Wnt mediator wingless-related MMTV integration site 7A (Wnt7a) activity is aberrant (see, e.g., Woods et al., 2006);
    • Odonto-onchyo-dermal hypoplasia, where Wnt mediator wingless related MMTV integration site 10a (Wnt10a) activity is aberrant (see, e.g., Adaimy et al., 2007); and
    • early onset obesity, where the Wnt mediator wingless related MMTV integration site 10b (Wnt10b) activity is aberrant (see, e.g., Christodoulides et al., 2006).


Moreover, aberrant telomerase protein component TERT expression and aberrant Wnt signalling are implicated in nephropathy, including HIV-associated nephropathy (see, e.g., Shkreli et al., 2011). Given the strong link between tankyrase inhibitors and modulation of both Wnt signalling and TERT function, it is likely that small molecule tankyrase inhibitors could be used in the treatment of these pathologies.


The inventors have identified a class of small molecule inhibitors of PARP superfamily members including PARP1 and Tankyrase 1 which are useful in the treatment of conditions, including proliferative conditions such as cancer. In some cases, these inhibitors are able to elicit biochemical inhibition of these targets as well as eliciting cellular activity including one or more or all of: (i) inhibition of Wnt signalling; (ii) inhibition of cell survival/proliferation; (iii) stabilisation of Axin and tankyrase levels; and (iv) formation of markers of DNA damage such as γH2AX foci.


It appears that the following 3-aryl-5-substituted-2H-isoquinolin-1-ones are known.














#
Structure
Registry No.







P01


embedded image


 70351-69-8





P02


embedded image


 70351-70-1





P03


embedded image


 70351-71-2





P04


embedded image


 70351-72-3





P05


embedded image


 203628-15-3





P06


embedded image


 203628-17-5





P07


embedded image


 203628-19-7





P08


embedded image


 220630-92-2





P09


embedded image


 223553-35-3





P10


embedded image


 884500-93-0





P11


embedded image


 884501-99-9





P12


embedded image


1256940-02-9





P13


embedded image


1256940-03-0





P14


embedded image


1256940-06-3





P15


embedded image


1256940-07-4





P16


embedded image


1256940-08-5





P17


embedded image


1256940-09-6





P18


embedded image


1256940-10-9





P19


embedded image


1256940-11-0





P20


embedded image


1256940-12-1





P21


embedded image


1256940-13-2





P22


embedded image


1256940-16-5





P23


embedded image


1256940-17-6





P24


embedded image


1262335-24-9









It appears that the following 3-aryl-5-unsubstituted-2H-isoquinolin-1-ones are known.














#
Structure
Registry No.







P25


embedded image


 19069-81-9





P26


embedded image


 98659-53-1





P27


embedded image


 98659-55-3





P28


embedded image


 145104-33-2





P29


embedded image


 223552-86-1





P30


embedded image


 223553-20-6





P31


embedded image


 376354-94-8





P32


embedded image


 376354-97-1





P33


embedded image


 503613-43-2





P34


embedded image


 503613-44-3





P35


embedded image


 630423-61-9





P36


embedded image


 630423-64-2





P37


embedded image


 721960-58-3





P38


embedded image


 721960-60-7





P39


embedded image


 721960-73-2





P40


embedded image


 862469-72-5





P41


embedded image


 924299-93-4





P42


embedded image


1044871-80-8





P43


embedded image


1044871-83-1





P44


embedded image


1193268-39-1





P45


embedded image


1193268-40-4





P46


embedded image


1253733-07-1





P47


embedded image


1253733-10-6





P48


embedded image


1417652-57-3









SUMMARY OF THE INVENTION

One aspect of the invention pertains to certain 3-aryl-5-substituted-2H-isoquinolin-1-one compounds (referred to herein as IQ compounds), as described herein.


Another aspect of the invention pertains to a composition (e.g., a pharmaceutical composition) comprising an IQ compound, as described herein, and a pharmaceutically acceptable carrier or diluent.


Another aspect of the invention pertains to a method of preparing a composition (e.g., a pharmaceutical composition) comprising the step of mixing an IQ compound, as described herein, and a pharmaceutically acceptable carrier or diluent.


Another aspect of the present invention pertains to a method of inhibiting PARP (e.g., PARP1, TNKS1, TNKS2, etc.) function (e.g., in a cell), in vitro or in vivo, comprising contacting the cell with an effective amount of an IQ compound, as described herein.


Another aspect of the present invention pertains to a method of inhibiting Wnt signalling (e.g., in a cell), in vitro or in vivo, comprising contacting the cell with an effective amount of an IQ compound, as described herein.


Another aspect of the present invention pertains to a method of treatment comprising administering to a subject in need of treatment a therapeutically-effective amount of an IQ compound, as described herein, preferably in the form of a pharmaceutical composition.


Another aspect of the present invention pertains to an IQ compound as described herein for use in a method of treatment of the human or animal body by therapy.


Another aspect of the present invention pertains to use of an IQ compound, as described herein, in the manufacture of a medicament for use in treatment.


In one embodiment, the treatment is treatment of a proliferative condition.


In one embodiment, the treatment is treatment of cancer.


In one embodiment, the treatment is treatment of head cancer; neck cancer; nervous system cancer; lung/mediastinum cancer; breast cancer; oesophagus cancer; stomach cancer; liver cancer; biliary tract cancer; pancreatic cancer; small bowel cancer; large bowel cancer; gynaecological cancer; genito-urinary cancer; thyroid gland cancer; adrenal gland cancer; skin cancer; bone sarcoma; soft tissue sarcoma; paediatric malignancy; Hodgkin's disease; non-Hodgkin's lymphoma; myeloma; leukaemia; or metastasis from an unknown primary site.


In one embodiment, the treatment is treatment of: a neurodegenerative disorder, such as multiple sclerosis (MS); a neurological disorder associated with demyelination; neonatal hypoxic ischemic encephalopathy (HIE); neonatal periventricular leukomalacia (PVL); a cardiac related pathology, such as myocardial infarction; cardiac damage (e.g., to repair cardiac damage); an infectious disease, such as a pathology related to Herpes Simplex Virus (HSV); a pathology related to Epstein-Barr Virus (EBV); a metabolic disease, such as a metabolic disease where glucose uptake is dysfunctional, such as diabetes, such as type 2 diabetes; or fibrosis (e.g., lung fibrosis).


In one embodiment, the treatment is treatment of: a neurodegenerative disorder, such as multiple sclerosis (MS); neonatal hypoxic ischemic encephalopathy (HIE); neonatal periventricular leukomalacia (PVL); a cardiac related pathology, such as myocardial infarction; a pathology related to Herpes Simplex Virus (HSV); a pathology related to Epstein-Barr Virus (EBV); or a metabolic disease such as type 2 diabetes.


In one embodiment, the treatment is treatment of: Alzheimer's disease; late onset Alzheimer's disease; Dupuytren skin disease; tooth agenesis; vascular defects in the eye; Osteoperosis-pseudoglioma Syndrome (OPPG); exudative vitreoretinopathy; familial exudative vitreoretinopathy; retinal angiogenesis; schizophrenia; osteoporosis; dermal hypoplasia; XX sex reversal; Mullerian-duct regression and virilization; SERKAL syndrome; anonychia; hyponychia; sclerosteosis; van Buchem disease; Fuhrmann syndrome; odonto-onchyo-dermal hypoplasia; Type 2 diabetes; obesity; early onset obesity; a nephropathy, such as HIV-associated nephropathy; early coronary disease; bone density defects; tetra-amelia syndrome; split-hand/foot malformation; caudal duplication; Fuhrmann syndrome; odonto-onycho-dermal dysplasia; skeletal dysplasia; focal dermal hypoplasia; autosomal recessive anonychia; or neural tube defects.


In one embodiment, the treatment is treatment of: Alzheimer's disease; Dupuytren skin disease; tooth agenesis; exudative vitreoretinopathy; schizophrenia; osteoporosis; dermal hypoplasia; XX sex reversal; anonychia; hyponychia; sclerosteosis; van Buchem disease; Fuhrmann syndrome; odonto-onchyo-dermal hypoplasia; early onset obesity; or a nephropathy, such as HIV-associated nephropathy.


Another aspect of the present invention pertains to a kit comprising (a) an IQ compound, as described herein, preferably provided as a pharmaceutical composition and in a suitable container and/or with suitable packaging; and (b) instructions for use, for example, written instructions on how to administer the compound.


Another aspect of the present invention pertains to an IQ compound obtainable by a method of synthesis as described herein, or a method comprising a method of synthesis as described herein.


Another aspect of the present invention pertains to an IQ compound obtained by a method of synthesis as described herein, or a method comprising a method of synthesis as described herein.


Another aspect of the present invention pertains to novel intermediates, as described herein, which are suitable for use in the methods of synthesis described herein.


Another aspect of the present invention pertains to the use of such novel intermediates, as described herein, in the methods of synthesis described herein.


As will be appreciated by one of skill in the art, features and preferred embodiments of one aspect of the invention will also pertain to other aspects of the invention.







DETAILED DESCRIPTION OF THE INVENTION
Compounds

One aspect of the present invention relates to certain compounds which are structurally related to 2H-isoquinolin-1-one.




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More particularly, the present invention relates to certain 3-aryl-5-substituted-2H-isoquinolin-1-one compounds, as defined herein.


Yet more particularly, the present invention relates to certain 2H-isoquinolin-1-one compounds which have both:

    • (a) a particular substituent (denoted herein as R5) at the 5-position; and
    • (b) a particular six-membered carboaryl or heteroaryl substituent (denoted herein as the ring containing W, X, Y, and Z) at the 3-position having a particular para-substituent (denoted herein as -L3P-R3N).


Thus, one aspect of the present invention pertains to compounds selected from compounds of the following formula, and pharmaceutically acceptable salts, N-oxides, hydrates, and solvates thereof, wherein —R3N, -L3P-, W, X, Y, Z, —R4, —R5, —R6, —R7, and —R8 are as defined herein (for convenience, collectively referred to herein as “3-aryl-5-substituted-2H-isoquinolin-1-one compounds” or “IQ compounds”):




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Some embodiments of the invention include the following:


(1) A compound selected from compounds of the following formula, and pharmaceutically acceptable salts, N-oxides, hydrates, and solvates thereof:




embedded image


wherein:

    • W is CRW, X is CRX, Y is CRY, and Z is CRZ (“phenyl”); or
    • W is N, X is CRX, Y is CRY, and Z is CRZ (“pyrid-2-yl”); or
    • W is CRW, X is N, Y is CRY, and Z is CRZ (“pyrid-3-yl”); or
    • W is N, X is CRX, Y is CRY, and Z is N (“pyrimidin-2-yl”); or
    • W is CRW, X is N, Y is N, and Z is CRZ (“pyrimidin-5-yl”); or
    • W is N, X is CRX, Y is N, and Z is CRZ (“pyrazin-2-yl”); or
    • W is N, X is N, Y is CRY, and Z is CRZ (“pyridazin-3-yl”);


      wherein:
    • —RW is independently —H or —RWW;
    • —RX is independently —H or —RXX;
      • —RY is independently —H or —RYY; and
    • —RZ is independently —H or —RZZ;


      wherein:
    • —RWW is independently —X1, —R1, —OH, —OR1, —CF3, or —OCF3;
    • —RXX is independently —X1, —R1, —OH, —OR1, —CF3, or —OCF3;
    • —RYY is independently —X1, —R1, —OH, —OR1, —CF3, or —OCF3; and
    • —RZZ is independently —X1, —R1, —OH, —OR1, —CF3, or —OCF3;


      wherein:
    • each —X1 is independently —F, —Cl, —Br, or —I; and
    • each —R1 is independently linear or branched saturated C1-4alkyl;


      and wherein:
    • -L3P- is independently a single covalent bond or -L3PL-;


      wherein:
    • -L3PL- is independently -L3PR1-, —C(═O)—, -L3PR2-C(═O)—, —S(═O)2—, -L3PR3-S(═O)2—, or —O-L3PR4-;
    • wherein:
    • each -L3PR1- is linear or branched saturated C1-4alkylene;
    • each -L3PR2- is linear or branched saturated C1-4alkylene;
    • each -L3PR3- is linear or branched saturated C1-4alkylene;
    • each -L3PR4- is linear or branched saturated C1-4alkylene;


      and wherein:
    • —R3N is independently —NH2, —NHRA, —NRARB, or —NRCRD;


      wherein:
    • each —RA is independently:
      • —RA1, —RA2, —RA3, —RA4, —RA5, -LA-RA2, -LA-RA3, -LA-RA4, or -LA-RA5;
    • each —RA1 is linear or branched saturated C1-6alkyl,
      • and is optionally substituted with one or more groups —RS1;
    • each —RA2 is saturated C3-6cycloalkyl,
      • and is optionally substituted with one or more groups —RS2C;
      • each —RA3 is non-aromatic C3-7heterocyclyl,
      • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen, if present, with a group —RSN;
    • each —RA4 is independently phenyl or naphthyl,
      • and is optionally substituted with one or more groups —RS3C;
    • each —RA5 is C6-10heteroaryl,
      • and is optionally substituted on carbon with one or more groups —RS3C,
      • and is optionally substituted on secondary nitrogen, if present, with a group —RSN;
    • each -LA- is linear or branched saturated C1-4alkylene;


      and wherein:
    • each —RS1 is independently:
      • —F, —Cl, —Br, —I,
      • —OH, —ORTT,
      • —OCF3,
      • —NH2, —NHRTT, —NRTT2, —RTM,
      • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
      • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —C(═O)RTM,
      • —NHC(═O)RTT, —NRTNC(═O)RTT,
      • —NHC(═O)NH2, —NHC(═O)NHRTT, —NHC(═O)NRTT2, —NHC(═O)RTM,
      • —NRTNC(═O)NH2, —NRTNC(═O)NHRTT, —NRTNC(═O)NRTT2, —NRTNC(═O)RTM,
      • —NHC(═O)ORTT, —NRTNC(═O)ORTT,
      • —OC(═O)NH2, —OC(═O)NHRTT, —OC(═O)NRTT2, —OC(═O)RTM,
      • —C(═O)RTT,
      • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
      • —NHS(═O)2RTT, —NRTNS(═O)2RTT,
      • —S(═O)2RTT,
      • —CN, —NO2, —SRTT, or ═O;
    • each —RS2C is independently:
      • —RTT,
      • —F, —Cl, —Br, —I,
      • —OH, —ORTT,
      • -LT-OH, -LT-ORTT,
      • —CF3, —OCF3,
      • —NH2, —NHRTT, —NRTT2, —RTM,
      • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
      • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
      • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —C(═O)RTM,
      • —NHC(═O)RTT, —NRTNC(═O)RTT,
      • —NHC(═O)NH2, —NHC(═O)NHRTT, —NHC(═O)NRTT2, —NHC(═O)RTM,
      • —NRTNC(═O)NH2, —NRTNC(═O)NHRTT, —NRTNC(═O)NRTT2, —NRTNC(═O)RTM,
      • —NHC(═O)ORTT, —NRTNC(═O)ORTT,
      • —OC(═O)NH2, —OC(═O)NHRTT, —OC(═O)NRTT2, —OC(═O)RTM,
      • —C(═O)RTT,
      • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
      • —NHS(═O)2RTT, —NRTNS(═O)2RTT,
      • —S(═O)2RTT,
      • —CN, —NO2, —SRTT, or ═O;
    • each —RS3C is independently:
      • —RTT,
      • —F, —Cl, —Br, —I,
      • —OH, —ORTT,
      • -LT-OH, -LT-ORTT,
      • —CF3, —OCF3,
      • —NH2, —NHRTT, —NRTT2, —RTM,
      • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
      • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
      • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —C(═O)RTM,
      • —NHC(═O)RTT, —NRTNC(═O)RTT,
      • —NHC(═O)NH2, —NHC(═O)NHRTT, —NHC(═O)NRTT2, —NHC(═O)RTM,
      • —NRTNC(═O)NH2, —NRTNC(═O)NHRTT, —NRTNC(═O)NRTT2, —NRTNC(═O)RTM, —NHC(═O)ORTT, —NRTNC(═O)ORTT,
      • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —OC(═O)RTM,
      • —C(═O)RTT,
      • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
      • —NHS(═O)2RTT, —NRTNS(═O)2RTT,
      • —S(═O)2RTT,
      • —CN, —NO2, or —SRTT;
      • and additionally, two adjacent groups —RS3C, if present, may together form: —O—CH2—O— or —O—CH2CH2—O—;
    • each —RSN is independently:
      • —RTT,
      • -LT-OH, -LT-ORTT,
      • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
      • —C(═O)RTT,
      • —C(═O)ORTT,
      • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —C(═O)RTM, or
      • —S(═O)2RTT;


        wherein:
    • each -LT- is linear or branched saturated C1-4alkylene;
    • each —RTT is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORTTT, wherein —RTTT is linear or branched saturated C1-4alkyl;
    • each —RTN is linear or branched saturated C1-4alkyl;
    • each —RTM is independently azetidino, pyrrolidino, piperidino, piperazino, morpholino, azepano, or diazepano, and is:
    • optionally substituted on carbon with one or more groups selected from: —RTMM, —C(═O)RTMM, —S(═O)2RTMM, —F, —NH2, —NHRTMM, —NRTMM2, —OH, and —ORTMM; and
    • optionally substituted on secondary nitrogen, if present, with a group selected from: —RTMM, —C(═O)RTMM, —C(═O)ORTMM, and —S(═O)2RTMM;
    • wherein each —RTMM is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl;


      and wherein:
    • —RB is independently —RB1, —RB2, or -LB-RB2;
    • —RB1 is linear or branched saturated C1-6alkyl, and is optionally substituted with —OH or —ORBB, wherein —RBB is linear or branched saturated C1-4alkyl;
    • —RB2 is saturated C3-6cycloalkyl; and
    • -LB- is linear or branched saturated C1-4alkylene;


      and wherein:
    • —NRCRD is independently —NRC1RD1, —NRC2RD2, —NRC3RD3, —NRC4RD4, or —NRC5RD5;


      wherein:
    • —NRC1RD1 is a monocyclic non-aromatic heterocyclyl group having from 4 to 8 ring atoms, wherein exactly 1 of said ring atoms is a ring heteroatom, and is N, or exactly 2 of said ring atoms are ring heteroatoms, and are both N, or exactly 2 of said ring atoms are ring heteroatoms, and are N and O, or exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2;
    • and wherein said monocyclic non-aromatic heterocyclyl group is:
    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN;
    • —NRC2RD2 is a fused bicyclic non-aromatic heterocyclyl group having from 7 to 12 ring atoms, wherein exactly 1 of said ring atoms is a ring heteroatom, and is N, or exactly 2 of said ring atoms are ring heteroatoms, and are both N, or exactly 2 of said ring atoms are ring heteroatoms, and are N and O, or exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2, or exactly 3 of said ring atoms are ring heteroatoms, one of which is N, and each of the other two is independently N, O, or S, wherein said S is optionally in the form of S(═O) or S(═O)2;
    • and wherein said fused bicyclic non-aromatic heterocyclyl group is:
    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN;
    • —NRC3RD3 is a bridged non-aromatic heterocyclyl group having from 7 to 11 ring atoms, wherein exactly 1 of said ring atoms is a ring heteroatom, and is N, or exactly 2 of said ring atoms are ring heteroatoms, and are both N, or exactly 2 of said ring atoms are ring heteroatoms, and are N and O, or exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2, or exactly 3 of said ring atoms are ring heteroatoms, one of which is N, and each of the other two is independently N, O, or S, wherein said S is optionally in the form of S(═O) or S(═O)2;
    • and wherein said bridged non-aromatic heterocyclyl group is:
    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN;
    • —NRC4RD4 is a spiro non-aromatic heterocyclyl group having from 6 to 12 ring atoms, wherein exactly 1 of said ring atoms is a ring heteroatom, and is N, or exactly 2 of said ring atoms are ring heteroatoms, and are both N, or exactly 2 of said ring atoms are ring heteroatoms, and are N and O, or exactly 2 of said ring atoms are ring heteroatoms, and are N and S, or exactly 3 of said ring atoms are ring heteroatoms, one of which is N, and each of the other two is independently N, O, or S, wherein said S is optionally in the form of S(═O) or S(═O)2;
    • and wherein said spiro non-aromatic heterocyclyl group is:
    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN;


      wherein:
    • each —RNC is independently:
      • —RQQ,
      • —F, —Cl, —Br, —I,
      • —OH, —ORQQ,
      • -LQ-OH, -LQ-ORQQ,
      • —CF3, —OCF3,
      • —NH2, —NHRQQ, —NRQQ2, —RQM,
      • -LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, -LQ-RQM,
      • —C(═O)OH, —C(═O)ORQQ, —OC(═O)RQQ,
      • —C(═O)NH2, —C(═O)NHRQQ, —C(═O)NRQQ2, —C(═O)RQM,
      • —NHC(═O)RQQ, —NRQNC(═O)RQQ,
      • —NHC(═O)NH2, —NHC(═O)NHRQQ, —NHC(═O)NRQQ2, —NHC(═O)RQM,
      • —NRQNC(═O)NH2, —NRQNC(═O)NHRQQ,
      • —NRQNC(═O)NRQQ2, —NRQNC(═O)RQM,
      • —NHC(═O)ORQQ, —NRQNC(═O)ORQQ,
      • —OC(═O)NH2, —OC(═O)NHRQQ, —OC(═O)NRQQ2, —OC(═O)RQM,
      • —C(═O)RQQ,
      • —S(═O)2NH2, —S(═O)2NHRQQ, —S(═O)2NRQQ2, —S(═O)2RQM,
      • —NHS(═O)2RQQ, —NRQNS(═O)2RQQ,
      • —S(═O)2RQQ,
      • —CN, —NO2, —SRQQ, or ═O;
    • each —RNN is independently:
      • —RQQ,
      • -LQ-OH, -LQ-ORQQ,
      • -LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, -LQ-RQM,
      • —C(═O)RQQ,
      • —C(═O)ORQQ,
      • —C(═O)NH2, —C(═O)NHRQQ, —C(═O)NRQQ2, —C(═O)RQM, or
      • —S(═O)2RQQ;


        wherein:
    • each -LQ- is linear or branched saturated C1-4alkylene;
    • each —RQQ is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl or benzyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORQQQ, and said phenyl and benzyl are optionally substituted with —RQQQ, wherein each —RQQQ is linear or branched saturated C1-4alkyl;
    • each —RQN is linear or branched saturated C1-4alkyl;
    • each —RQM is independently azetidino, pyrrolidino, piperidino, piperazino, morpholino, azepano, or diazepano, and is:
    • optionally substituted on carbon with one or more groups selected from: —RQMM, —C(═O)RQMM, —S(═O)2RQMM, —F, —NH2, —NHRQMM, —NRQMM2, —OH, and —ORQMM; and
    • optionally substituted on secondary nitrogen, if present, with a group selected from: —RQMM, —C(═O)RQMM, —C(═O)ORQMM, and —S(═O)2RQMM;
    • wherein each —RQMM is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl;


      and wherein:
    • —NRC5RD5 is independently: 1H-pyrrol-1-yl; 2H-isoindol-2-yl; 1H-indol-1-yl; 1H-pyrazol-1-yl; 1H-benzoimidazol-1-yl; 1H-imidazol-1-yl; 2H-indazol-2-yl; 1H-indazol-1-yl; 4H-[1,2,4]triazol-4-yl; 1H-[1,2,3]triazol-1-yl; 1H-[1,2,4]triazol-1-yl; 1H-benzotriazol-1-yl; or 1H-tetrazol-1-yl; and is optionally substituted with one or more groups —RH;


      wherein each —RH is independently:
    • —RHH,
    • —F, —Cl, —Br, —I,
    • —OH, —ORHH,
    • -LH-OH, -LH-ORHH,
    • —CF3, —OCF3,
    • —NH2, —NHRHH, —NRHH2, —RHM,
    • -LH-NH2, -LH-NHRHH, -LH-NRHH2, -LH-RHM,
    • —C(═O)OH, —C(═O)ORHH, —OC(═O)RHH,
    • —C(═O)NH2, —C(═O)NHRHH, —C(═O)NRHH2, —C(═O)RHM,
    • —NHC(═O)RHH, —NRHNC(═O)RHH,
    • —NHC(═O)NH2, —NHC(═O)NHRHH, —NHC(═O)NRHH2, —NHC(═O)RHM,
    • —NRHNC(═O)NH2, —NRHNC(═O)NHRHH, —NRHNC(═O)NRHH2, —NRHNC(═O)RHM,
    • —NHC(═O)ORHH, —NRHNC(═O)ORHH,
    • —OC(═O)NH2, —OC(═O)NHRHH, —OC(═O)NRHH2, —OC(═O)RHM,
    • —C(═O)RHH,
    • —S(═O)2NH2, —S(═O)2NHRHH, —S(═O)2NRHH2, —S(═O)2RHM,
    • —NHS(═O)2RHH, —NRHNS(═O)2RHH,
    • —S(═O)2RHH,
    • —CN, —NO2, or —SRHH;


      wherein:
    • each -LH- is linear or branched saturated C1-4alkylene;
    • each —RHH is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORHHH, wherein —RHHH is linear or branched saturated C1-4alkyl;
    • each —RHN is linear or branched saturated C1-4alkyl;
    • each —RHM is independently azetidino, pyrrolidino, piperidino, piperazino, morpholino, azepano, or diazepano, and is:
    • optionally substituted on carbon with one or more groups selected from: —RHMM, —C(═O)RHMM, —S(═O)2RHMM, —F, —NH2, —NHRHMM, —NRHMM2, —OH, and —ORHMM; and
    • optionally substituted on secondary nitrogen, if present, with a group selected from: —RHMM, —C(═O)RHMM, —C(═O)ORHMM, and —S(═O)2RHMM;
    • wherein each —RHMM is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl;


      and wherein:
    • —R5 is independently —R5A, —R5B, —R5C, —R5D, or —R5E;
    • —R5A is linear or branched saturated C1-4alkyl;
    • —R5B is saturated C3-6cycloalkyl;
    • —R5C is independently —F, —Cl, —Br, or —I;
    • —R5D is —CF3; and
    • —R5E is independently —C≡CH or C3-6alkynyl optionally substituted with one or more groups —REE; wherein each —REE is independently selected from —OH, —ORBEE, —NH2, —NHREEE, and —NREEE2; wherein each —REEE is linear or branched saturated C1-4alkyl;


      and wherein:
    • —R4 is —H;
    • —R6 is independently —H or —F; and
    • —R7 is independently —H or —F; and
    • —R8 is independently —H or —F.


For the avoidance of doubt, it is not intended that any two or more of —R3N, -L3P, W, X, Y, Z, —R4, —R5, —R6, —R7, and —R8 together form a ring fused to the ring(s) to which they are attached. For example, it is not intended that —R4 and —R5 together form a ring fused to the ring to which they are attached. Similarly, it is not intended that —R4 and Z together form a ring fused to the rings to which they are attached. Similarly, it is not intended that —R4 and W together form a ring fused to the rings to which they are attached.


For the avoidance of doubt, the phrase “substituent on carbon” is intended to refer to a substituent which is attached to a carbon ring atom. Similarly, the phrase “substituent on secondary nitrogen” is intended to refer to a substituent which is attached to a nitrogen ring atom which, in the absence of the substituent, would be a secondary nitrogen ring atom (i.e., —NH—). Consequently, a pyridyl group may only have “substituents on carbon”, whereas 1H-pyrrole may have both “substituents on carbon” and a “substituent on secondary nitrogen”, as illustrated below.




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Similarly, a piperidino group may only have “substituents on carbon”, whereas piperizino may have both “substituents on carbon” and a “substituent on secondary nitrogen”, as illustrated below.




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The Groups W, X, Y, and Z

(2) A compound according to (1), wherein:

    • W is CRW, X is CRX, Y is CRY, and Z is CRZ (“phenyl”); or
    • W is N, X is CRX, Y is CRY, and Z is CRZ (“pyrid-2-yl”); or
    • W is CRW, X is N, Y is CRY, and Z is CRZ (“pyrid-3-yl”); or
    • W is N, X is CRX, Y is CRY, and Z is N (“pyrimidin-2-yl”); or
    • W is CRW, X is N, Y is N, and Z is CRZ (“pyrimidin-5-yl”).


(3) A compound according to any (1), wherein:

    • W is CRW, X is CRX, Y is CRY, and Z is CRZ (“phenyl”); or
    • W is CRW, X is N, Y is CRY, and Z is CRZ (“pyrid-3-yl”); or
    • W is CRW, X is N, Y is N, and Z is CRZ (“pyrimidin-5-yl”).


(4) A compound according to (1), wherein:

    • W is CRW, X is CRX, Y is CRY, and Z is CRZ (“phenyl”).


(5) A compound according to (1), wherein:

    • W is CRW, X is N, Y is CRY, and Z is CRZ (“pyrid-3-yl”).


(6) A compound according to (1), wherein:

    • W is CRW, X is N, Y is N, and Z is CRZ (“pyrimidin-5-yl”).


The Group —RW

(7) A compound according to any one of (1) to (6), wherein —RW, if present, is —H.


(8) A compound according to any one of (1) to (6), wherein —RW, if present, is —RWW.


The Group —RX

(9) A compound according to any one of (1) to (8), wherein —RX, if present, is —H.


(10) A compound according to any one of (1) to (8), wherein —RX, if present, is —RXX.


The Group —RY

(11) A compound according to any one of (1) to (10), wherein —RY, if present, is —H.


(12) A compound according to any one of (1) to (10), wherein —RY, if present, is —RYY.


The Group —RZ

(13) A compound according to any one of (1) to (12), wherein —RZ, if present, is —H.


(14) A compound according to any one of (1) to (12), wherein —RZ, if present, is —RZZ.


The Group —RWW

(15) A compound according to any one of (1) to (14), wherein —RWW, if present, is independently —X1, —R1, or —CF3.


(16) A compound according to any one of (1) to (14), wherein —RWW, if present, is independently —X1 or —R1.


(17) A compound according to any one of (1) to (14), wherein —RWW, if present, is independently —X1.


(18) A compound according to any one of (1) to (14), wherein —RWW, if present, is independently —R1.


The Group —RXX

(19) A compound according to any one of (1) to (18), wherein —RXX, if present, is independently —X1, —R1, or —CF3.


(20) A compound according to any one of (1) to (18), wherein —RXX, if present, is independently —X1 or —R1.


(21) A compound according to any one of (1) to (18), wherein —RXX, if present, is independently —X1.


(22) A compound according to any one of (1) to (18), wherein —RXX, if present, is independently —R1.


The Group —RYY

(23) A compound according to any one of (1) to (22), wherein —RYY, if present, is independently —X1, —R1, or —CF3.


(24) A compound according to any one of (1) to (22), wherein —RYY, if present, is independently —X1 or —R1.


(25) A compound according to any one of (1) to (22), wherein —RYY, if present, is independently —X1.


(26) A compound according to any one of (1) to (22), wherein —RYY, if present, is independently —R1.


The Group —RZZ

(27) A compound according to any one of (1) to (26), wherein —RZZ, if present, is independently —X1, —R1, or —CF3.


(28) A compound according to any one of (1) to (26), wherein —RZZ, if present, is independently —X1 or —R1.


(29) A compound according to any one of (1) to (26), wherein —RZZ, if present, is independently —X1.


(30) A compound according to any one of (1) to (26), wherein —RZZ, if present, is independently —R1.


The Group —X1

(31) A compound according to any one of (1) to (30), wherein each —X1, if present, is independently —F, —Cl, or —Br.


(32) A compound according to any one of (1) to (30), wherein each —X1, if present, is independently —F or —Cl.


(33) A compound according to any one of (1) to (30), wherein each —X1, if present, is —F.


(34) A compound according to any one of (1) to (30), wherein each —X1, if present, is —Cl.


(35) A compound according to any one of (1) to (30), wherein each —X1, if present, is —Br.


(36) A compound according to any one of (1) to (30), wherein each —X1, if present, is —I.


The Group —R1

(37) A compound according to any one of (1) to (36), wherein each —R1, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(38) A compound according to any one of (1) to (36), wherein each —R1, if present, is independently -Me, -Et, -nPr, or -iPr.


(39) A compound according to any one of (1) to (36), wherein each —R1, if present, is independently -Me or -Et.


(40) A compound according to any one of (1) to (36), wherein each —R1, if present, is -Me.


The Group -L3P-

(41) A compound according to any one of (1) to (40), wherein -L3P- is a single covalent bond.


(42) A compound according to any one of (1) to (40), wherein -L3P- is -L3PL-.


The Group -L3PL-

(43) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is independently -L3PR1-, —C(═O)—, -L3PR2-C(═O)—, —O-L3PR4-, or —S(═O)2—.


(44) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is independently -L3PR1-, —C(═O)—, —O-L3PR4-, or —S(═O)2—.


(45) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is -L3PR1-.


(46) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is —C(═O)—.


(47) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is -L3PR2-C(═O)—.


(48) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is —S(═O)2—.


(49) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is -L3PR3-S(═O)2—.


(50) A compound according to any one of (1) to (42), wherein -L3PL-, if present, is —O-L3PR4-.


The Group -L3PR1-

(51) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(52) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(53) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(54) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(55) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(56) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —CH2— or —CH2CH2—.


(57) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is —CH2—.


(58) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —CH(Me)-.


(59) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is independently —C(Me)2-.


(60) A compound according to any one of (1) to (50), wherein each -L3PR1-, if present, is —CH2CH2—.


The Group -L3PR2-

(61) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(62) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(63) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(64) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(65) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(66) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —CH2— or —CH2CH2—.


(67) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is —CH2—.


(68) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —CH(Me)-.


(69) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is independently —C(Me)2-.


(70) A compound according to any one of (1) to (60), wherein each -L3PR2-, if present, is —CH2CH213 .


The Group -L3PR3-

(71) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(72) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(73) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(74) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(75) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(76) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is independently —CH2— or —CH2CH2—.


(77) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is —CH2—.


(78) A compound according to any one of (1) to (70), wherein each -L3PR3-, if present, is —CH2CH2—.


The Group -L3PR4-

(79) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(80) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(81) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(82) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(83) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(84) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is independently —CH2— or —CH2CH2—.


(85) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is —CH2—.


(86) A compound according to any one of (1) to (78), wherein each -L3PR4-, if present, is —CH2CH2—.


The Group —R3N

(87) A compound according to any one of (1) to (86), wherein —R3N is independently —NHRA, —NRARB, or —NRCRD.


(88) A compound according to any one of (1) to (86), wherein —R3N is independently —NRARB or —NRCRD.


(89) A compound according to any one of (1) to (86), wherein —R3N is —NH2.


(90) A compound according to any one of (1) to (86), wherein —R3N is —NHRA.


(91) A compound according to any one of (1) to (86), wherein —R3N is —NRARB.


(92) A compound according to any one of (1) to (86), wherein —R3N is —NRCRd.


The Group —RA

(93) A compound according to any one of (1) to (92), wherein each —RA, if present, is independently: —RA1, —RA2, —RA3, -LA-RA2, or -LA-RA3.


(94) A compound according to any one of (1) to (92), wherein each —RA, if present, is independently: —RA1, —RA3, or -LA-RA3.


(95) A compound according to any one of (1) to (92), wherein each —RA, if present, is —RA1.


(96) A compound according to any one of (1) to (92), wherein each —RA, if present, is —RA2.


(97) A compound according to any one of (1) to (92), wherein each —RA, if present, is —RA3.


(98) A compound according to any one of (1) to (92), wherein each —RA, if present, is —RA4.


(99) A compound according to any one of (1) to (92), wherein each —RA, if present, is —RA5.


(100) A compound according to any one of (1) to (92), wherein each —RA, if present, is -LA-RA2.


(101) A compound according to any one of (1) to (92), wherein each —RA, if present, is -LA-RA3.


(102) A compound according to any one of (1) to (92), wherein each —RA, if present, is -LA-RA4.


(103) A compound according to any one of (1) to (92), wherein each —RA, if present, is LA-RA5.


The Group —RA1

(104) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently linear or branched saturated C1-4alkyl, and is optionally substituted with one or more groups —RS1.


(105) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently linear or branched saturated C1-4alkyl, and is optionally substituted with one or more groups selected from: —OH, —ORTT, —NH2, —NHRTT, and —NRTT2.


(106) A compound according to any one of (1) to (xx), wherein each —RA1, if present, is independently linear or branched saturated C1-4alkyl, and is optionally substituted with one or more groups selected from: —OH and —ORTT.


(107) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu, and is optionally substituted with one or more groups —RS1.


(108) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu, and is optionally substituted with one or more groups selected from: —OH, —ORTT, —NH2, —NHRTT, and —NRTT2.


(109) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me, -Et, -nPr, or -iPr, and is optionally substituted with one or more groups —RS1.


(110) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me, -Et, -nPr, or -iPr, and is optionally substituted with one or more groups selected from: —OH, —ORTT, —NH2, —NHRTT, and —NRTT2.


(111) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me or -Et, and is optionally substituted with one or more groups —RS1.


(112) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently linear or branched saturated C1-4alkyl.


(113) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(114) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me, -Et, -nPr, or -iPr.


(115) A compound according to any one of (1) to (103), wherein each —RA1, if present, is independently -Me or -Et.


(116) A compound according to any one of (1) to (103), wherein each —RA1, if present, is -Me.


The Group —RA2

(117) A compound according to any one of (1) to (116), wherein each —RA2, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, and is optionally substituted with one or more groups —RS2C.


(118) A compound according to any one of (1) to (116), wherein each —RA2, if present, is independently cyclopropyl, cyclobutyl, or cyclopentyl, and is optionally substituted with one or more groups —RS2C.


(119) A compound according to any one of (1) to (116), wherein each —RA2, if present, is independently cyclopropyl or cyclobutyl, and is optionally substituted with one or more groups —RS2C.


The Group —RA3

(120) A compound according to any one of (1) to (119), wherein each —RA3, if present, is independently oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, dioxanyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl, or diazepanyl,

    • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen, if present, with a group —RSN.


(121) A compound according to any one of (1) to (119), wherein each —RA3, if present, is independently tetrahydrofuranyl, tetrahydropyranyl, dioxanyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl,

    • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen, if present, with a group —RSN.


(122) A compound according to any one of (1) to (119), wherein each —RA3, if present, is independently tetrahydropyranyl or piperidinyl,

    • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen, if present, with a group —RSN.


(123) A compound according to any one of (1) to (119), wherein each —RA3, if present, is tetrahydropyranyl, and is optionally substituted on carbon with one or more groups —RS2c.


(124) A compound according to any one of (1) to (119), wherein each —RA3, if present, is piperidinyl,

    • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen with a group —RSN.


(125) A compound according to any one of (1) to (119), wherein each —RA3, if present, is pyrrolidinyl,

    • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen with a group —RSN.


(126) A compound according to any one of (1) to (119), wherein each —RA3, if present, is azetidinyl,

    • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen with a group —RSN.


The Group —RA4

(127) A compound according to any one of (1) to (126), wherein each —RA4, if present, is phenyl, and is optionally substituted with one or more groups —RS3C.


(128) A compound according to any one of (1) to (126), wherein each —RA4, if present, is naphthyl, and is optionally substituted with one or more groups —RS3C.


The Group —RA5

(129) A compound according to any one of (1) to (128), wherein each —RA5, if present, is independently furanyl, thienyl, pyrrolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, benzoimidazolyl, indazolyl, benzofuranyl, benzothienyl, benzooxazolyl, benzothiazolyl, benzoisoxazolyl, benzoisothiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, quinazolinyl, or phthalazinyl,

    • and is optionally substituted on carbon with one or more groups —RS3C,
    • and is optionally substituted on secondary nitrogen, if present, with a group —RSN.


(130) A compound according to any one of (1) to (128), wherein each —RA5, if present, is independently furanyl, thienyl, pyrrolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, or pyrazinyl,

    • and is optionally substituted on carbon with one or more groups —RS3C,
    • and is optionally substituted on secondary nitrogen, if present, with a group —RSN.


(131) A compound according to any one of (1) to (128), wherein each —RA5, if present, is independently furanyl, thienyl, pyrrolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, isoxazolyl, or isothiazolyl,

    • and is optionally substituted on carbon with one or more groups —RS3C,
    • and is optionally substituted on secondary nitrogen, if present, with a group —RSN.


(132) A compound according to any one of (1) to (128), wherein each —RA5, if present, is imidazolyl,

    • and is optionally substituted on carbon with one or more groups —RS2C,
    • and is optionally substituted on secondary nitrogen with a group —RSN.


(133) A compound according to any one of (1) to (128), wherein each —RA5, if present, is independently pyridyl, pyridazinyl, pyrimidinyl, or pyrazinyl,

    • and is optionally substituted on carbon with one or more groups —RS3C.


The Group -LA-

(134) A compound according to any one of (1) to (133), wherein each -LA-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(135) A compound according to any one of (1) to (133), wherein each -LA-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(136) A compound according to any one of (1) to (133), wherein each -LA-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(137) A compound according to any one of (1) to (133), wherein each -LA-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(138) A compound according to any one of (1) to (133), wherein each -LA-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(139) A compound according to any one of (1) to (133), wherein each -LA-, if present, is independently —CH2— or —CH2CH2—.


(140) A compound according to any one of (1) to (133), wherein each -LA-, if present, is —CH2—.


(141) A compound according to any one of (1) to (133), wherein each -LA-, if present, is —CH2CH2—.


The Group —RS1

(142) A compound according to any one of (1) to (141), wherein each —RS1, if present, is independently:

    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —C(═O)RTM,
    • —C(═O)RTT,
    • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
    • —NHS(═O)2RTT, —NRTNS(═O)2RTT,
    • —S(═O)2RTT,
    • —CN, —NO2, —SRTT, or ═O.


(143) A compound according to any one of (1) to (141), wherein each —RS1, if present, is independently:

    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —C(═O)RTM,
    • —NHC(═O)RTT, —NRTNC(═O)RTT,
    • —C(═O)RTT,
    • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
    • —NHS(═O)2RTT, —NRTNS(═O)2RTT, or
    • —S(═O)2RTT.


(144) A compound according to any one of (1) to (141), wherein each —RS1, if present, is independently:

    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, C(═O)RTM,
    • —NHC(═O)RTT, —NRTNC(═O)RTT, or
    • —C(═O)RTT.


(145) A compound according to any one of (1) to (141), wherein each —RS1, if present, is independently:

    • —F,
    • —OH, —ORTT,
    • —NH2, —NHRTT, —NRTT2, or —RTM.


(146) A compound according to any one of (1) to (141), wherein each —RS1, if present, is independently:

    • —OH, —ORTT,
    • —NH2, —NHRTT, —NRTT2, or —RTM.


(147) A compound according to any one of (1) to (141), wherein each —RS1, if present, is independently —OH or —ORTT.


The Group —RS2C

(148) A compound according to any one of (1) to (148), wherein each —RS2C, if present, is independently:

    • —RTT,
    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —CF3, —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
    • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, C(═O)RTM,
    • —NHC(═O)RTT, —NRTNC(═O)RTT,
    • —C(═O)RTT,
    • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
    • —NHS(═O)2RTT, —NRTNS(═O)2RTT,
    • —S(═O)2RTT,
    • —CN, —NO2, —SRTT, or ═O.


(149) A compound according to any one of (1) to (148), wherein each —RS2C, if present, is independently:

    • —RTT,
    • —F, —Cl, —Br, —I,


—OH, —ORTT,


-LT-OH, -LT-ORTT,


—CF3, —OCF3,


—NH2, —NHRTT, —NRTT2, —RTM,


-LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,


—C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,

    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, O(═O)RTM,
    • —NHC(═O)RTT, —NRTNC(═O)RTT,
    • —C(═O)RTT,
    • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
    • —NHS(═O)2RTT, —NRTNS(═O)2RTT,
    • —S(═O)2RTT, or
    • ═O.


(150) A compound according to any one of (1) to (148), wherein each —RS2C, if present, is independently:

    • —RTT,
    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM, or
    • ═O.


(151) A compound according to any one of (1) to (148), wherein each —RS2C, if present, is independently:

    • —RTT,
    • —F,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM, or
    • ═O.


(152) A compound according to any one of (1) to (148), wherein each —RS2C, if present, is independently:

    • —RTT,
    • —F,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM, or
    • ═O.


(153) A compound according to any one of (1) to (148), wherein each —RS2C, if present, is independently:

    • —RTT,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM, or


═O.


(154) A compound according to any one of (1) to (148), wherein each —RS2C, if present, is independently:

    • —RTT,
    • —OH, —ORTT,
    • —NH2, —NHRTT, —NRTT2, —RTM, or
    • ═O.


The Group —RS3C

(155) A compound according to any one of (1) to (154), wherein each —RS3C, if present, is independently:

    • —RTT,
    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —CF3, —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
    • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, —C(═O)RTM,
    • —NHC(═O)RTT, —NRTNC(═O)RTT,
    • —C(═O)RTT,
    • —S(═O)2NH2, —S(═O)2NHRTT, —S(═O)2NRTT2, —S(═O)2RTM,
    • —NHS(═O)2RTT, —NRTNS(═O)2RTT,
    • —S(═O)2RTT,
    • —CN, —NO2, or —SRTT.


(156) A compound according to any one of (1) to (154), wherein each —RS3C, if present, is independently:

    • —RTT,
    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —CF3, —OCF3,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
    • —C(═O)OH, —C(═O)ORTT, —OC(═O)RTT,
    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, C(═O)RTM,
    • —NHC(═O)RTT, —NRTNC(═O)RTT, or
    • —C(═O)RTT.


(157) A compound according to any one of (1) to (154), wherein each —RS3C, if present, is independently:

    • —RTT,
    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • -LT-OH, -LT-ORTT,
    • —NH2, —NHRTT, —NRTT2, —RTM,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, or -LT-RTM.


(158) A compound according to any one of (1) to (154), wherein each —RS3C, if present, is independently:

    • —RTT,
    • —F, —Cl, —Br, —I,
    • —OH, —ORTT,
    • —NH2, —NHRTT, —NRTT2, or —RTM.


The Group —RSN

(159) A compound according to any one of (1) to (158), wherein each —RSN, if present, is independently:

    • —RTT,
    • -LT-OH, -LT-ORTT,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
    • —C(═O)RTT,
    • —C(═O)ORTT,
    • —C(═O)NH2, —C(═O)NHRTT, —C(═O)NRTT2, or —C(═O)RTM.


(160) A compound according to any one of (1) to (158), wherein each —RSN, if present, is independently:

    • —RTT,
    • -LT-OH, -LT-ORTT,
    • -LTNH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM,
    • —C(═O)RTT, or
    • —C(═O)ORTT.


(161) A compound according to any one of (1) to (158), wherein each —RSN, if present, is independently:

    • —RTT,
    • -LT-OH, -LT-ORTT,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, -LT-RTM, or
    • —C(═O)RTT.


(162) A compound according to any one of (1) to (158), wherein each —RSN, if present, is independently:

    • —RTT,
    • -LT-OH, -LT-ORTT,
    • -LT-NH2, -LT-NHRTT, -LT-NRTT2, or
    • —C(═O)RTT.


(163) A compound according to any one of (1) to (158), wherein each —RSN, if present, is independently:

    • —RTT,
    • —C(═O)RTT, or
    • —C(═O)ORTT.


(164) A compound according to any one of (1) to (158), wherein each —RSN, if present, is independently —RTT or —C(═O)RTT.


(165) A compound according to any one of (1) to (158), wherein each —RSN, if present, is independently —RTT.


The Group -LT-

(166) A compound according to any one of (1) to (165), wherein each -LT-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(167) A compound according to any one of (1) to (165), wherein each -LT-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(168) A compound according to any one of (1) to (165), wherein each -LT-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(169) A compound according to any one of (1) to (165), wherein each -LT-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(170) A compound according to any one of (1) to (165), wherein each -LT-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(171) A compound according to any one of (1) to (165), wherein each -LT-, if present, is independently —CH2— or —CH2CH2—.


(172) A compound according to any one of (1) to (165), wherein each -LT-, if present, is —CH2—.


(173) A compound according to any one of (1) to (165), wherein each -LT-, if present, is —CH2CH2—.


The Group —RTT

(174) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl.


(175) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, phenyl, or benzyl.


(176) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently linear or branched saturated C1-4alkyl, phenyl, or benzyl.


(177) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORTTT, wherein —RTTT is linear or branched saturated C1-4alkyl.


(178) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl.


(179) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORTTT, wherein —RTTT is linear or branched saturated C1-4alkyl.


(180) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl.


(181) A compound according to any one of (1) to (173), wherein each —RTT, if present, is linear or branched saturated C1-4alkyl, and is optionally substituted with —OH or —ORTTT, wherein —RTTT is linear or branched saturated C1-4alkyl.


(182) A compound according to any one of (1) to (173), wherein each —RTT, if present, is linear or branched saturated C1-4alkyl.


(183) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(184) A compound according to any one of (1) to (173), wherein each —RTT, if present, is independently -Me or -tBu.


(185) A compound according to any one of (1) to (173), wherein each —RTT, if present, is -Me.


(186) A compound according to any one of (1) to (173), wherein each —RTT, if present, is -tBu.


The Group —RTTT

(187) A compound according to any one of (1) to (186), wherein each —RTTT, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(188) A compound according to any one of (1) to (186), wherein each —RTTT, if present, is independently -Me or -Et.


(189) A compound according to any one of (1) to (186), wherein each —RTTT, if present, is -Me.


The Group —RTN

(190) A compound according to any one of (1) to (189), wherein each —RTN, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(191) A compound according to any one of (1) to (189), wherein each —RTN, if present, is independently -Me or -Et.


(192) A compound according to any one of (1) to (189), wherein each —RTN, if present, is -Me.


The Group —RTM

(193) A compound according to any one of (1) to (192), wherein each —RTM, if present, is independently pyrrolidino, piperidino, piperazino, or morpholino, and is:

    • optionally substituted on carbon with one or more groups selected from: —RTMM, —C(═O)RTMM, —S(═O)2RTMM, —F, —NH2, —NHRTMM, —NRTMM2, —OH, and —ORTMM; and
    • optionally substituted on secondary nitrogen, if present, with a group selected from: —RTMM, —C(═O)RTMM, —C(═O)ORTMM, and —S(═O)2RTMM;
    • wherein each —RTMM is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl.


The Group —RTMM

(194) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, phenyl, or benzyl.


(195) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently linear or branched saturated C1-4alkyl, phenyl, or benzyl.


(196) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl.


(197) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl.


(198) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is linear or branched saturated C1-4alkyl.


(199) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(200) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently -Me or -Et.


(201) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is -Me.


(202) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently saturated C3-6cycloalkyl.


(203) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.


(204) A compound according to any one of (1) to (193), wherein each —RTMM, if present, is cyclopropyl.


The Group —RB

(205) A compound according to any one of (1) to (204), wherein —RB, if present, is —RB1.


(206) A compound according to any one of (1) to (204), wherein —RB, if present, is —RB2.


(207) A compound according to any one of (1) to (204), wherein —RB, if present, is -LB-RB2.


The Group —RB1

(208) A compound according to any one of (1) to (207), wherein —RB1, if present, is linear or branched saturated C1-6alkyl.


(209) A compound according to any one of (1) to (207), wherein —RB1, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu, and is optionally substituted with —OH or —ORBB, wherein —RBB is linear or branched saturated C1-4alkyl.


(210) A compound according to any one of (1) to (207), wherein —RB1, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(211) A compound according to any one of (1) to (207), wherein —RB1, if present, is independently -Me, -Et, -nPr, or -iPr.


(212) A compound according to any one of (1) to (207), wherein —RB1, if present, is independently: -Me; or -Et that is optionally substituted with —OH or —ORBB, wherein —RBB is linear or branched saturated C1-4alkyl.


(213) A compound according to any one of (1) to (207), wherein —RB1, if present, is independently -Me, -Et, —CH2CH2OH, or —CH2CH2OMe.


(214) A compound according to any one of (1) to (207), wherein —RB1, if present, is independently -Me, -Et, or —CH2CH2OH.


(215) A compound according to any one of (1) to (207), wherein —RB1, if present, is independently -Me or -Et.


(216) A compound according to any one of (1) to (207), wherein —RB1, if present, is -Me.


The Group —RBB

(217) A compound according to any one of (1) to (216), wherein —RBB, if present, is independently -Me or -Et.


(218) A compound according to any one of (1) to (216), wherein —RBB, if present, is -Me.


The Group —RB2

(219) A compound according to any one of (1) to (218), wherein —RB2, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.


(220) A compound according to any one of (1) to (218), wherein —RB2, if present, is independently cyclopropyl, cyclobutyl, or cyclopentyl.


(221) A compound according to any one of (1) to (218), wherein —RB2, if present, is independently cyclopropyl or cyclobutyl.


(222) A compound according to any one of (1) to (218), wherein —RB2, if present, is cyclopropyl.


The Group -LB-

(223) A compound according to any one of (1) to (222), wherein each -LB-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(224) A compound according to any one of (1) to (222), wherein each -LB-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et), or —CH2CH2—.


(225) A compound according to any one of (1) to (222), wherein each -LB-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(226) A compound according to any one of (1) to (222), wherein each -LB-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(227) A compound according to any one of (1) to (222), wherein each -LB-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(228) A compound according to any one of (1) to (222), wherein each -LB-, if present, is independently —CH2— or —CH2CH2—.


(229) A compound according to any one of (1) to (222), wherein each -LB-, if present, is —CH2—.


(230) A compound according to any one of (1) to (222), wherein each -LB-, if present, is —CH2CH2—.


The Group —NRCRD

(231) A compound according to any one of (1) to (230), wherein —NRCRD, if present, is —NRC1RD1.


(232) A compound according to any one of (1) to (230), wherein —NRCRD, if present, is —NRC2RD2.


(233) A compound according to any one of (1) to (230), wherein —NRCRD, if present, is —NRC3RD3.


(234) A compound according to any one of (1) to (230), wherein —NRCRD, if present, is —NRC4RD4.


(235) A compound according to any one of (1) to (230), wherein —NRCRD, if present, is —NRC5RD5.


The Group —NRC1RD1

(236) A compound according to any one of (1) to (235), wherein —NRC1RD1, if present, is a monocyclic non-aromatic heterocyclyl group having from 4 to 7 ring atoms.


(237) A compound according to any one of (1) to (235), wherein —NRC1RD1, if present, is a monocyclic non-aromatic heterocyclyl group having from 5 to 7 ring atoms.


(238) A compound according to any one of (1) to (235), wherein —NRC1RD1, if present, is a monocyclic non-aromatic heterocyclyl group having 5 ring atoms.


(239) A compound according to any one of (1) to (235), wherein —NRC1RD1, if present, is a monocyclic non-aromatic heterocyclyl group having 6 ring atoms.


(240) A compound according to any one of (1) to (235), wherein —NRC1R1, if present, is a monocyclic non-aromatic heterocyclyl group having 7 ring atoms.


(241) A compound according to any one of (1) to (235), wherein, in —NRC1RD1, if present, exactly 1 of said ring atoms is a ring heteroatom, and is N.


(242) A compound according to any one of (1) to (235), wherein, in —NRC1RD1, if present, exactly 2 of said ring atoms are ring heteroatoms, and are both N.


(243) A compound according to any one of (1) to (235), wherein, in —NRC1RD1, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and 0.


(244) A compound according to any one of (1) to (235), wherein, in —NRC1RD1, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2.


(245) A compound according to any one of (1) to (235), wherein, in —NRC1RD1, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S.


(246) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is independently selected from the following groups, wherein S, if present, is optionally in the form of S(═O) or S(═O)2, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN:




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(247) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN:




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(248) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN:




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(249) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is the following group, and is optionally substituted on carbon with one or more groups —RNC:




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(250) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is the following group, and is optionally substituted on carbon with one or more groups —RNC:




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(251) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is the following group, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen with a group —RNN:




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(252) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is the following group, and is optionally substituted on carbon with one or more groups —RNC:




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(253) A compound according to any one of (1) to (235), wherein, —NRC1RD1, if present, is the following group, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen with a group —RNN:




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The Group —NRC2RD2

(254) A compound according to any one of (1) to (253), wherein —NRC2RD2, if present, is a fused bicyclic non-aromatic heterocyclyl group having from 7 to 12 ring atoms, wherein exactly 1 of said ring atoms is a ring heteroatom, and is N, or exactly 2 of said ring atoms are ring heteroatoms, and are both N, or exactly 2 of said ring atoms are ring heteroatoms, and are N and O, or exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2;

    • and wherein said fused bicyclic non-aromatic heterocyclyl group is:
    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN.


(255) A compound according to any one of (1) to (253), wherein —NRC2RD2, if present, is a fused bicyclic non-aromatic heterocyclyl group having from 8 to 10 ring atoms.


(256) A compound according to any one of (1) to (253), wherein —NRC2RD2, if present, is a fused bicyclic non-aromatic heterocyclyl group having 8 ring atoms.


(257) A compound according to any one of (1) to (253), wherein —NRC2RD2, if present, is a fused bicyclic non-aromatic heterocyclyl group having 9 ring atoms.


(258) A compound according to any one of (1) to (253), wherein —NRC2RD2, if present, is a fused bicyclic non-aromatic heterocyclyl group having 10 ring atoms.


(259) A compound according to any one of (1) to (258), wherein, in —NRC2RD2, if present, exactly 1 of said ring atoms is a ring heteroatom, and is N.


(260) A compound according to any one of (1) to (258), wherein, in —NRC2RD2, if present, exactly 2 of said ring atoms are ring heteroatoms, and are both N.


(261) A compound according to any one of (1) to (258), wherein, in —NRC2RD2, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and O.


(262) A compound according to any one of (1) to (258), wherein, in —NRC2RD2, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2.


(263) A compound according to any one of (1) to (258), wherein, in —NRC2RD2, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S.


(264) A compound according to any one of (1) to (253), wherein, —NRC2RD2, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN:




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(265) A compound according to any one of (1) to (253), wherein, —NRC2RD2, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN:




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(266) A compound according to any one of (1) to (253), wherein, —NRC2RD2, if present, is the following group, and is optionally substituted on carbon with one or more groups —RNC:




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(267) A compound according to any one of (1) to (253), wherein, —NRC2RD2, if present, is the following group, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen with a group —RNN:




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The Group —NRC3RD3

(268) A compound according to any one of (1) to (267), wherein —NRC3RD3, if present, is a bridged non-aromatic heterocyclyl group having from 7 to 11 ring atoms, wherein exactly 1 of said ring atoms is a ring heteroatom, and is N, or exactly 2 of said ring atoms are ring heteroatoms, and are both N, or exactly 2 of said ring atoms are ring heteroatoms, and are N and O;

    • and wherein said bridged non-aromatic heterocyclyl group is:
    • optionally substituted on carbon with one or more groups —RNc, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN.


(269) A compound according to any one of (1) to (267), wherein —NRC3RD3, if present, is a bridged non-aromatic heterocyclyl group having 7 ring atoms.


(270) A compound according to any one of (1) to (267), wherein —NRC3RD3, if present, is a bridged non-aromatic heterocyclyl group having 8 ring atoms.


(271) A compound according to any one of (1) to (267), wherein —NRC3RD3, if present, is a bridged non-aromatic heterocyclyl group having 9 ring atoms.


(272) A compound according to any one of (1) to (267), wherein —NRC3RD3, if present, is a bridged non-aromatic heterocyclyl group having 11 ring atoms.


(273) A compound according to any one of (1) to (272), wherein, in —NRC3RD3, if present, exactly 1 of said ring atoms is a ring heteroatom, and is N.


(274) A compound according to any one of (1) to (272), wherein, in —NRC3RD3, if present, exactly 2 of said ring atoms are ring heteroatoms, and are both N.


(275) A compound according to any one of (1) to (272), wherein, in —NRC3RD3, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and O.


(276) A compound according to any one of (1) to (272), wherein, in —NRC3RD3, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2.


(277) A compound according to any one of (1) to (272), wherein, in —NRC3RD3, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S.


(278) A compound according to any one of (1) to (272), wherein, in —NRC3RD3, if present, exactly 3 of said ring atoms are ring heteroatoms, one of which is N, and each of the other two is independently N, O, or S, wherein said S is optionally in the form of S(═O) or S(═O)2.


(279) A compound according to any one of (1) to (272), wherein, in —NRC3RD3, if present, exactly 3 of said ring atoms are ring heteroatoms, one of which is N, and each of the other two is independently N, O, or S.


(280) A compound according to any one of (1) to (267), wherein, —NRC3RD3, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with groups —RNN:




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(281) A compound according to any one of (1) to (267), wherein, —NRC3RD3, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with groups —RNN:




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(282) A compound according to any one of (1) to (267), wherein, —NRC3RD3, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNc, and
    • optionally substituted on secondary nitrogen, if present, with groups —RNN:




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(283) A compound according to any one of (1) to (267), wherein, —NRC3RD3, if present, is the following group, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen with a group —RNN:




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(284) A compound according to any one of (1) to (267), wherein, —NRC3RD3, if present, is the following group, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen with groups —RNN:




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The Group —NRC4RD4

(285) A compound according to any one of (1) to (284), wherein —NRC4RD4, if present, is a spiro non-aromatic heterocyclyl group having 7 ring atoms.


(286) A compound according to any one of (1) to (284), wherein —NRC4RD4, if present, is a spiro non-aromatic heterocyclyl group having 8 ring atoms.


(287) A compound according to any one of (1) to (284), wherein —NRC4RD4, if present, is a spiro non-aromatic heterocyclyl group having 9 ring atoms.


(288) A compound according to any one of (1) to (284), wherein —NRC4RD4, if present, is a spiro non-aromatic heterocyclyl group having 10 ring atoms.


(289) A compound according to any one of (1) to (284), wherein —NRC4RD4, if present, is a spiro non-aromatic heterocyclyl group having 11 ring atoms.


(290) A compound according to any one of (1) to (284), wherein —NRC4RD4, if present, is a spiro non-aromatic heterocyclyl group having 12 ring atoms.


(291) A compound according to any one of (1) to (290), wherein, in —NRC4RD4, if present, exactly 1 of said ring atoms is a ring heteroatom, and is N.


(292) A compound according to any one of (1) to (290), wherein, in —NRC4RD4, if present, exactly 2 of said ring atoms are ring heteroatoms, and are both N.


(293) A compound according to any one of (1) to (290), wherein, in —NRC4RD4, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and O.


(294) A compound according to any one of (1) to (290), wherein, in —NRC4RD4, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S, wherein said S is optionally in the form of S(═O) or S(═O)2.


(295) A compound according to any one of (1) to (290), wherein, in —NRC4RD4, if present, exactly 2 of said ring atoms are ring heteroatoms, and are N and S.


(296) A compound according to any one of (1) to (290), wherein, in —NRC4RD4, if present, exactly 3 of said ring atoms are ring heteroatoms, one of which is N, and each of the other two is independently N, O, or S, wherein said S is optionally in the form of S(═O) or S(═O)2.


(297) A compound according to any one of (1) to (290), wherein, in —NRC4RD4, if present, exactly 3 of said ring atoms are ring heteroatoms, one of which is N, and each of the other two is independently N, O, or S.


(298) A compound according to any one of (1) to (284), wherein, —NRC4RD4, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN:




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(299) A compound according to any one of (1) to (284), wherein, —NRC4RD4, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen, if present, with a group —RNN:




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(300) A compound according to any one of (1) to (284), wherein, —NRC4RD4, if present, is independently selected from the following groups, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen with a group —RNN:




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(301) A compound according to any one of (1) to (284), wherein, —NRC4RD4, if present, is the following group, and is:

    • optionally substituted on carbon with one or more groups —RNC, and
    • optionally substituted on secondary nitrogen with a group —RNN:




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The Group —RNC

(302) A compound according to any one of (1) to (301), wherein each —RNC, if present, is independently:

    • —RQQ
    • —F, —Cl, —Br, —I,
    • —OH, —ORQQ,
    • -LQ-OH, -LQ-ORQQ,
    • —NH2, —NHRQQ, —NRQQ2, —RQM,
    • -LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, -LQ-RQM, or
    • ═O.


(303) A compound according to any one of (1) to (301), wherein each —RNC, if present, is independently:

    • —RQQ,
    • —OH, —ORQQ,
    • —NH2, —NHRQQ, —NRQQ2, —RQM, or
    • ═O.


(304) A compound according to any one of (1) to (301), wherein each —RNC, if present, is independently —RQQ.


The Group —RNN

(305) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently:

    • —RQQ,
    • -LQ-OH, -LQ-ORQQ,
    • -LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, -LQ-RQM,
    • —C(═O)RQQ,
    • —C(═O)ORQQ,
    • —C(═O)NH2, —C(═O)NHRQQ, —C(═O)NRQQ2, or —C(═O)RQM.


(306) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently:

    • —RQQ,
    • -LQ-OH, -LQ-ORQQ,
    • -LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, -LQ-RQM,
    • —C(═O)RQQ, or
    • —C(═O)ORQQ.


(307) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently:

    • —RQQ,
    • -LQ-OH, -LQ-ORQQ,
    • -LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, -LQ-RQM, or
    • —C(═O)RQQ.


(308) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently:

    • —RQQ,
    • -LQ-OH, -LQ-ORQQ,
    • -LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, or
    • —C(═O)RQQ.


(309) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently:

    • —RQQ,
    • —C(═O)RQQ, or
    • —C(═O)ORQQ.


(310) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently —RQQ or —C(═O)RQQ.


(311) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently —RQQ.


(312) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently: —RQQ, -LQ-OH, or -LQ-ORQQ.


(313) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently: -LQ-OH or -LQ-ORQQ.


(314) A compound according to any one of (1) to (304), wherein each —RNN, if present, is independently: -LQ-OH.


The Group -LQ-

(315) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(316) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(317) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(318) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(319) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(320) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is independently —CH2— or —CH2CH2—.


(321) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is —CH2—.


(322) A compound according to any one of (1) to (314), wherein each -LQ-, if present, is —CH2CH2—.


The Group —RQQ

(323) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORQQQ, wherein —RQQQ is linear or branched saturated C1-4alkyl.


(324) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl.


(325) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, phenyl, or benzyl.


(326) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl, phenyl, or benzyl.


(327) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORQQ, wherein —RQQ is linear or branched saturated C1-4alkyl.


(328) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl.


(329) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORQQ, wherein —RQQ is linear or branched saturated C1-4alkyl.


(330) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl.


(331) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is linear or branched saturated C1-4alkyl, and is optionally substituted with —OH or —ORQQ, wherein —RQQ is linear or branched saturated C1-4alkyl.


(332) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is linear or branched saturated C1-4alkyl.


(333) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(334) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently -Me or -tBu.


(335) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is -Me.


(336) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is -tBu.


(337) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is saturated C3-6cycloalkyl.


(338) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.


(339) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is cyclopropyl.


(340) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is saturated C3-6cycloalkyl-methyl.


(341) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is independently cyclopropyl-methyl, cyclobutyl-methyl, cyclopentyl-methyl, or cyclohexyl-methyl.


(342) A compound according to any one of (1) to (322), wherein each —RQQ, if present, is cyclopropyl-methyl.


The Group —RQQQ

(343) A compound according to any one of (1) to (342), wherein each —RQQQ, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(344) A compound according to any one of (1) to (342), wherein each —RQQQ, if present, is independently -Me or -Et.


(345) A compound according to any one of (1) to (342), wherein each —RQQQ, if present, is independently -Me.


The Group —RQN

(346) A compound according to any one of (1) to (345), wherein each —RQN, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(347) A compound according to any one of (1) to (345), wherein each —RQN, if present, is independently -Me or -Et.


(348) A compound according to any one of (1) to (345), wherein each —RQN, if present, is independently -Me.


The Group —RQM

(349) A compound according to any one of (1) to (348), wherein each —RQM, if present, is independently pyrrolidino, piperidino, piperazino, or morpholino, and is:

    • optionally substituted on carbon with one or more groups selected from: —RQMM, —C(═O)RQMM, —S(═O)2RQMM, —F, —NH2, —NHRQMM, —NRQMM2, —OH, and —ORQMM; and
    • optionally substituted on secondary nitrogen, if present, with a group selected from: —RQMM, —C(═O)RQMM, —C(═O)ORQMM, and —S(═O)2RQMM;
    • wherein each —RQMM is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl.


The Group —RQMM

(350) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, phenyl, or benzyl.


(351) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently linear or branched saturated C1-4alkyl, phenyl, or benzyl.


(352) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl.


(353) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl.


(354) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is linear or branched saturated C1-4alkyl.


(355) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(356) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently -Me or -Et.


(357) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is -Me.


(358) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently saturated C3-6cycloalkyl.


(359) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.


(360) A compound according to any one of (1) to (349), wherein each —RQMM, if present, is cyclopropyl.


The Group —NRC5RD5

(361) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is independently: 1H-pyrrol-1-yl; 2H-isoindol-2-yl; 1H-indol-1-yl; 1H-pyrazol-1-yl; 1H-benzoimidazol-1-yl; 1H-imidazol-1-yl; 2H-indazol-2-yl; 1H-indazol-1-yl; 4H-[1,2,4]triazol-4-yl; 1H-[1,2,3]triazol-1-yl; 1H-[1,2,4]triazol-1-yl; 1H-benzotriazol-1-yl; or 1H-tetrazol-1-yl; and is optionally substituted with one or more groups —RH.




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(362) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is independently: 1H-pyrrol-1-yl; 1H-pyrazol-1-yl; 1H-imidazol-1-yl; 4H-[1,2,4]triazol-4-yl; 1H-[1,2,3]triazol-1-yl; 1H-[1,2,4]triazol-1-yl; or 1H-tetrazol-1-yl; and is optionally substituted with one or more groups —RH.


(363) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is independently: 1H-pyrrol-1-yl; 1H-pyrazol-1-yl; or 1H-imidazol-1-yl; and is optionally substituted with one or more groups —RH.


(364) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is 1H-pyrrol-1-yl; and is optionally substituted with one or more groups —RH.


(365) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is 1H-pyrazol-1-yl; and is optionally substituted with one or more groups —RH.


(366) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is 1H-imidazol-1-yl; and is optionally substituted with one or more groups —RH.


(367) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is 1H-[1,2,4]triazol-1-yl; and is optionally substituted with one or more groups —RH.


(368) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is 1H-benzoimidazol-1-yl; and is optionally substituted with one or more groups —RH.


(369) A compound according to any one of (1) to (360), wherein —NRC5RD5, if present, is 1H-indol-1-yl; and is optionally substituted with one or more groups —RH.


The Group —RH

(370) A compound according to any one of (1) to (369), wherein each —RH, if present, is independently:

    • —RHH,
    • —F, —Cl, —Br, —I,
    • —OH, —ORHH,
    • -LH-OH, -LH-ORHH,
    • —CF3, —OCF3,
    • —NH2, —NHRHH, —NRHH2, —RTM,
    • -LH-NH2, -LH-NHRHH, -LH-NRHH2, -LH-RHM,
    • —C(═O)OH, —C(═O)ORHH, —OC(═O)RHH,
    • —C(═O)NH2, —C(═O)NHRHH, —C(═O)NRHH2, —C(═O)RHM,
    • —NHC(═O)RHH, —NRHNC(═O)RHH, or
    • —C(═O)RHH.


(371) A compound according to any one of (1) to (369), wherein each —RH, if present, is independently:

    • —RHH,
    • —F, —Cl, —Br, —I,
    • —OH, —ORHH,
    • -LH-OH, -LHH-ORHH,
    • —NH2, —NHRHH, —NRHH2, —RHM,
    • -LH-NH2, -LH-NHRHH, -LH-NRHH2, or -LH-RHM.


(372) A compound according to any one of (1) to (369), wherein each —RH, if present, is independently:

    • —RHH,
    • —OH, —ORHH,
    • —NH2, —NHRHH, —NRHH2, or —RHM.


(373) A compound according to any one of (1) to (369), wherein each —RH, if present, is independently —RHH.


The Group -LH-

(374) A compound according to any one of (1) to (373), wherein each -LH-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH2CH2—, —CH(Me)CH2—, —CH2CH(Me)—, —C(Me)2CH2—, —CH2C(Me)2-, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(375) A compound according to any one of (1) to (373), wherein each -LH-, if present, is independently —CH2—, —CH(Me)—, —C(Me)2-, —CH(Et)-, or —CH2CH2—.


(376) A compound according to any one of (1) to (373), wherein each -LH-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.


(377) A compound according to any one of (1) to (373), wherein each -LH-, if present, is independently —CH2—, —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(378) A compound according to any one of (1) to (373), wherein each -LH-, if present, is independently —CH2CH2—, —CH2CH2CH2—, or —CH2CH2CH2CH2—.


(379) A compound according to any one of (1) to (373), wherein each -LH-, if present, is independently —CH2— or —CH2CH2—.


(380) A compound according to any one of (1) to (373), wherein each -LH-, if present, is —CH2—.


(381) A compound according to any one of (1) to (373), wherein each -LH-, if present, is —CH2CH2—.


The Group —RHH

(382) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl.


(383) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, phenyl, or benzyl.


(384) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently linear or branched saturated C1-4alkyl, phenyl, or benzyl.


(385) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORHHH, wherein —RHHH is linear or branched saturated C1-4alkyl.


(386) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl.


(387) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl; wherein said linear or branched saturated C1-4alkyl is optionally substituted with —OH or —ORHHH, wherein —RQQ is linear or branched saturated C1-4alkyl.


(388) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl.


(389) A compound according to any one of (1) to (381), wherein each —RHH, if present, is linear or branched saturated C1-4alkyl, and is optionally substituted with —OH or —ORHHH, wherein —RHHH is linear or branched saturated C1-4alkyl.


(390) A compound according to any one of (1) to (381), wherein each —RHH, if present, is linear or branched saturated C1-4alkyl.


(391) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(392) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently -Me or -tBu.


(393) A compound according to any one of (1) to (381), wherein each —RHH, if present, is -Me.


(394) A compound according to any one of (1) to (381), wherein each —RHH, if present, is -tBu.


(395) A compound according to any one of (1) to (381), wherein each —RHH, if present, is saturated C3-6cycloalkyl.


(396) A compound according to any one of (1) to (381), wherein each —RHH, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.


(397) A compound according to any one of (1) to (381), wherein each —RHH, if present, is cyclopropyl.


The Group —RHHH

(398) A compound according to any one of (1) to (397), wherein each —RHHH, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(399) A compound according to any one of (1) to (397), wherein each —RHHH, if present, is independently -Me or -Et.


(400) A compound according to any one of (1) to (397), wherein each —RHH, if present, is independently -Me.


The Group —RHN

(401) A compound according to any one of (1) to (400), wherein each —RHN, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(402) A compound according to any one of (1) to (400), wherein each —RHN, if present, is independently -Me or -Et.


(403) A compound according to any one of (1) to (400), wherein each —RHN, if present, is independently -Me.


The Group —RHM

(404) A compound according to any one of (1) to (403), wherein each —RHM, if present, is independently pyrrolidino, piperidino, piperazino, or morpholino, and is:

    • optionally substituted on carbon with one or more groups selected from: —RHMM, —C(═O)RHMM, —S(═O)2RHMM, —F, —NH2, —NHRHMM, —NRHMM2, —OH, and —ORHMM; and
    • optionally substituted on secondary nitrogen, if present, with a group selected from: —RHMM, —C(═O)RHMM, —C(═O)ORHMM, and —S(═O)2RHMM;
    • wherein each —RHMM is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, saturated C3-6cycloalkyl-methyl, phenyl, or benzyl.


The Group -RHMM

(405) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, phenyl, or benzyl.


(406) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently linear or branched saturated C1-4alkyl, phenyl, or benzyl.


(407) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl.


(408) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently linear or branched saturated C1-4alkyl or saturated C3-6cycloalkyl.


(409) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is linear or branched saturated C1-4alkyl.


(410) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(411) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently -Me or -Et.


(412) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is -Me.


(413) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently saturated C3-6cycloalkyl.


(414) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.


(415) A compound according to any one of (1) to (404), wherein each —RHMM, if present, is cyclopropyl.


The Group —R5

(416) A compound according to any one of (1) to (415), wherein —R5 is independently —R5A, —R5B, —R5C, or —R5D.


(417) A compound according to any one of (1) to (415), wherein —R5 is —R5A.


(418) A compound according to any one of (1) to (415), wherein —R5 is —R5B.


(419) A compound according to any one of (1) to (415), wherein —R5 is —R5C.


(420) A compound according to any one of (1) to (415), wherein —R5 is —R5D.


(421) A compound according to any one of (1) to (415), wherein —R5 is —R5E.


The Group —R5A

(422) A compound according to any one of (1) to (421), wherein —R5A, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(423) A compound according to any one of (1) to (421), wherein —R5A, if present, is independently -Me, -Et, -nPr, or -iPr.


(424) A compound according to any one of (1) to (421), wherein —R5A, if present, is independently -Me or -Et.


(425) A compound according to any one of (1) to (421), wherein —R5A, if present, is -Me.


The Group —R5B

(426) A compound according to any one of (1) to (425), wherein —R5B, if present, is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.


(427) A compound according to any one of (1) to (425), wherein —R5B, if present, is independently cyclopropyl, cyclobutyl, or cyclopentyl.


(428) A compound according to any one of (1) to (425), wherein —R5B, if present, is independently cyclopropyl or cyclobutyl.


(429) A compound according to any one of (1) to (425), wherein —R5B, if present, is cyclopropyl.


The Group —R5C

(430) A compound according to any one of (1) to (429), wherein —R5C, if present, is independently —F, —Cl, or —Br.


(431) A compound according to any one of (1) to (429), wherein —R5C, if present, is independently —F or —Cl.


(432) A compound according to any one of (1) to (429), wherein —R5C, if present, is —F.


(433) A compound according to any one of (1) to (429), wherein —R5C, if present, is —Cl.


(434) A compound according to any one of (1) to (429), wherein —R5C, if present, is —Br.


(435) A compound according to any one of (1) to (429), wherein —R5C, if present, is —I.


The Group —R5E

(436) A compound according to any one of (1) to (435), wherein —R5E, if present, is independently —C≡CH or C3-4alkynyl optionally substituted with one or more groups —REE; wherein each —REE is independently selected from —OH, —OREEE, —NH2, —NHREEE, and —NREEE2; wherein each —REEE is linear or branched saturated C1-4alkyl.


(437) A compound according to any one of (1) to (435), wherein —R5E, if present, is —C≡CH.


(438) A compound according to any one of (1) to (435), wherein —R5E, if present, is C3-4alkynyl optionally substituted with one or more groups —REE; wherein each —REE is independently selected from —OH, —OREEE, —NH2, —NHREEE, and —NREEE2; wherein each —REEE is linear or branched saturated C1-4alkyl.


(439) A compound according to any one of (1) to (435), wherein —R5E, if present, is independently —C≡CH, —C≡CH—CH3, —C≡CH—CH2REE, —C≡CH—CH2CH3 or —C≡CH—CH2CH2REE; wherein each —REE is independently selected from —OH, —OREEE, —NH2, —NHREEE, and —NREEE2; wherein each —REEE is linear or branched saturated C1-4alkyl.


(440) A compound according to any one of (1) to (435), wherein —R5E, if present, is independently —C≡CH—CH3 or —C≡CH—CH2REE; wherein each —REE is independently selected from —OH, —OREEE, —NH2, —NHREEE, and —NREEE2; wherein each —REEE is linear or branched saturated C1-4alkyl.


(441) A compound according to any one of (1) to (435), wherein —R5E, if present, is independently —C≡CH—CH2CH3 or —C≡CH—CH2CH2REE; wherein each —REE is independently selected from —OH, —OREEE, —NH2, —NHREEE, and —NREEE2; wherein each —REEE is linear or branched saturated C1-4alkyl.


The Group —REE

(442) A compound according to any one of (1) to (441), wherein —REE, if present, is independently —OH or —OREEE.


(443) A compound according to any one of (1) to (441), wherein —REE, if present, is independently —NH2, —NHREEE, or —NREEE2.


The Group —REEE

(444) A compound according to any one of (1) to (443), wherein each —REEE, if present, is independently -Me, -Et, -nPr, -iPr, -nBu, -iBu, or -tBu.


(445) A compound according to any one of (1) to (443), wherein each —REEE, if present, is independently -Me, -Et, -nPr, or -iPr.


(446) A compound according to any one of (1) to (443), wherein each —REEE, if present, is independently -Me or -Et.


(447) A compound according to any one of (1) to (443), wherein each —REEE, if present, is -Me.


The Group —R6

(448) A compound according to any one of (1) to (447), wherein —R6 is —H.


(449) A compound according to any one of (1) to (447), wherein —R6 is —F.


The Group —R7

(450) A compound according to any one of (1) to (449), wherein —R7 is —H.


(451) A compound according to any one of (1) to (449), wherein —R7 is —F.


The Group —R8

(452) A compound according to any one of (1) to (451), wherein —R8 is —H.


(453) A compound according to any one of (1) to (451), wherein —R8 is —F.


Specific Compounds

(454) A compound according to (1), selected from compounds of the following formulae and pharmaceutically acceptable salts, N-oxides, hydrates, and solvates thereof:













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Combinations

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the chemical groups represented by the variables (e.g., W, X, Y, Z, —RW, —RX, —RY, —RZ, —RWW, —RXX, —RYY, —RZZ, —X1, —R1, -L3P-, -L3PL-, -L3PR1-, -L3PR2-, -L3PR3-, -L3PR4-, —R3N, —RA, —RA1, —RA2, —RA3, —RA4, —RA5, -LA-, —RS1, —RS2C, —RS3C, —RSN, -LT-, —RTT, —RTTT, —RTN, —RTM, —RTMM, —RB, —RB1, —RB2, -LB-, —RBB, —NRCRD, —NRC1RD1, —NRC2RD2, —NRC3RD3, —NRC4RD4, —NRC5RD5, —RNC, —RNN, -LQ-, —RQQ, —RQN, —RQM, —RQMM, —RH, -LH-, —RHH, —RHN, —RHM, —RHMM, —R5, —R5A, —R5B, —R5C, —R5D, —R5E, —R5EE, —R5EEE, —R4, —R6, —R7, —R8, etc.) are specifically embraced by the present invention and are disclosed herein just as if each and every combination was individually and explicitly disclosed, to the extent that such combinations embrace compounds that are stable compounds (i.e., compounds that can be isolated, characterised, and tested for biological activity). In addition, all sub-combinations of the chemical groups listed in the embodiments describing such variables are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub-combination of chemical groups was individually and explicitly disclosed herein.


Substantially Purified Forms

One aspect of the present invention pertains to IQ compounds, as described herein, in substantially purified form and/or in a form substantially free from contaminants.


In one embodiment, the compound is in substantially purified form and/or in a form substantially free from contaminants.


In one embodiment, the compound is in a substantially purified form with a purity of least 50% by weight, e.g., at least 60% by weight, e.g., at least 70% by weight, e.g., at least 80% by weight, e.g., at least 90% by weight, e.g., at least 95% by weight, e.g., at least 97% by weight, e.g., at least 98% by weight, e.g., at least 99% by weight.


Unless specified, the substantially purified form refers to the compound in any stereoisomeric or enantiomeric form. For example, in one embodiment, the substantially purified form refers to a mixture of stereoisomers, i.e., purified with respect to other compounds. In one embodiment, the substantially purified form refers to one stereoisomer, e.g., optically pure stereoisomer. In one embodiment, the substantially purified form refers to a mixture of enantiomers. In one embodiment, the substantially purified form refers to an equimolar mixture of enantiomers (i.e., a racemic mixture, a racemate). In one embodiment, the substantially purified form refers to one enantiomer, e.g., optically pure enantiomer.


In one embodiment, the compound is in a form substantially free from contaminants wherein the contaminants represent no more than 50% by weight, e.g., no more than 40% by weight, e.g., no more than 30% by weight, e.g., no more than 20% by weight, e.g., no more than 10% by weight, e.g., no more than 5% by weight, e.g., no more than 3% by weight, e.g., no more than 2% by weight, e.g., no more than 1% by weight.


Unless specified, the contaminants refer to other compounds, that is, other than stereoisomers or enantiomers. In one embodiment, the contaminants refer to other compounds and other stereoisomers. In one embodiment, the contaminants refer to other compounds and the other enantiomer.


In one embodiment, the compound is in a substantially purified form with an optical purity of at least 60% (i.e., 60% of the compound, on a molar basis, is the desired stereoisomer or enantiomer, and 40% is undesired stereoisomer(s) or enantiomer), e.g., at least 70%, e.g., at least 80%, e.g., at least 90%, e.g., at least 95%, e.g., at least 97%, e.g., at least 98%, e.g., at least 99%.


Isomers

Certain compounds may exist in one or more particular geometric, optical, enantiomeric, diasteriomeric, epimeric, atropic, stereoisomeric, tautomeric, conformational, or anomeric forms, including but not limited to, cis- and trans-forms; E- and Z-forms; c-, t-, and r-forms; endo- and exo-forms; R-, S-, and meso-forms; D- and L-forms; d- and l-forms; (+) and (−) forms; keto-, enol-, and enolate-forms; syn- and anti-forms; synclinal- and anticlinal-forms; α- and β-forms; axial and equatorial forms; boat-, chair-, twist-, envelope-, and halfchair-forms; and combinations thereof, hereinafter collectively referred to as “isomers” (or “isomeric forms”).


Note that, except as discussed below for tautomeric forms, specifically excluded from the term “isomers,” as used herein, are structural (or constitutional) isomers (i.e., isomers which differ in the connections between atoms rather than merely by the position of atoms in space). For example, a reference to a methoxy group, —OCH3, is not to be construed as a reference to its structural isomer, a hydroxymethyl group, —CH2OH. Similarly, a reference to ortho-chlorophenyl is not to be construed as a reference to its structural isomer, meta-chlorophenyl. However, a reference to a class of structures may well include structurally isomeric forms falling within that class (e.g., C1-7alkyl includes n-propyl and iso-propyl; butyl includes n-, iso-, sec-, and tert-butyl; methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).


The above exclusion does not pertain to tautomeric forms, for example, keto-, enol-, and enolate-forms, as in, for example, the following tautomeric pairs: keto/enol (illustrated below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime, thioketone/enethiol, N-nitroso/hydroxyazo, and nitro/aci-nitro.




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For example, 1H-pyridin-2-one-5-yl and 2-hydroxyl-pyridin-5-yl (shown below) are tautomers of one another. A reference herein to one is intended to encompass both.




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Note that specifically included in the term “isomer” are compounds with one or more isotopic substitutions. For example, H may be in any isotopic form, including 1H, 2H (D), and 3H (T); C may be in any isotopic form, including 12C, 13C, and 14C; O may be in any isotopic form, including 16O and 18O; and the like.


Unless otherwise specified, a reference to a particular compound includes all such isomeric forms, including mixtures (e.g., racemic mixtures) thereof. Methods for the preparation (e.g., asymmetric synthesis) and separation (e.g., fractional crystallisation and chromatographic means) of such isomeric forms are either known in the art or are readily obtained by adapting the methods taught herein, or known methods, in a known manner.


Salts

It may be convenient or desirable to prepare, purify, and/or handle a corresponding salt of the compound, for example, a pharmaceutically-acceptable salt. Examples of pharmaceutically acceptable salts are discussed in Berge et al., 1977, “Pharmaceutically Acceptable Salts,” J. Pharm. Sci., Vol. 66, pp. 1-19.


For example, if the compound is anionic, or has a functional group which may be anionic (e.g., —COOH may be —COO−), then a salt may be formed with a suitable cation. Examples of suitable inorganic cations include, but are not limited to, alkali metal ions such as Na+ and K+, alkaline earth cations such as Ca2+ and Mg2+, and other cations such as Al3+. Examples of suitable organic cations include, but are not limited to, ammonium ion (i.e., NH4+) and substituted ammonium ions (e.g., NH3R+, NH2R2+, NHR3+, NR4+). Examples of some suitable substituted ammonium ions are those derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, as well as amino acids, such as lysine and arginine. An example of a common quaternary ammonium ion is N(CH3)4+.


If the compound is cationic, or has a functional group which may be cationic (e.g., —NH2 may be —NH3+), then a salt may be formed with a suitable anion. Examples of suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids: hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, and phosphorous.


Examples of suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetyoxybenzoic, acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic, ethanesulfonic, formic, fumaric, glucheptonic, gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalene carboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic, succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examples of suitable polymeric organic anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethyl cellulose.


Unless otherwise specified, a reference to a particular compound also includes salt forms thereof.


N-Oxides

It may be convenient or desirable to prepare, purify, and/or handle a corresponding N-oxide of the compound. For example, a compound having a pyridyl group may be prepared, purified, and/or handled as the corresponding N-oxide.




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Unless otherwise specified, a reference to a particular compound also includes N-oxide forms thereof.


Hydrates and Solvates

It may be convenient or desirable to prepare, purify, and/or handle a corresponding solvate of the compound. The term “solvate” is used herein in the conventional sense to refer to a complex of solute (e.g., compound, salt of compound) and solvent. If the solvent is water, the solvate may be conveniently referred to as a hydrate, for example, a mono-hydrate, a di-hydrate, a tri-hydrate, etc.


Unless otherwise specified, a reference to a particular compound also includes solvate and hydrate forms thereof.


Chemically Protected Forms

It may be convenient or desirable to prepare, purify, and/or handle the compound in a chemically protected form. The term “chemically protected form” is used herein in the conventional chemical sense and pertains to a compound in which one or more reactive functional groups are protected from undesirable chemical reactions under specified conditions (e.g., pH, temperature, radiation, solvent, and the like). In practice, well known chemical methods are employed to reversibly render unreactive a functional group, which otherwise would be reactive, under specified conditions. In a chemically protected form, one or more reactive functional groups are in the form of a protected or protecting group (also known as a masked or masking group or a blocked or blocking group). By protecting a reactive functional group, reactions involving other unprotected reactive functional groups can be performed, without affecting the protected group; the protecting group may be removed, usually in a subsequent step, without substantially affecting the remainder of the molecule. See, for example, Protective Groups in Organic Synthesis (T. Greene and P. Wuts; 4th Edition; John Wiley and Sons, 2006).


A wide variety of such “protecting,” “blocking,” or “masking” methods are widely used and well known in organic synthesis. For example, a compound which has two nonequivalent reactive functional groups, both of which would be reactive under specified conditions, may be derivatized to render one of the functional groups “protected,” and therefore unreactive, under the specified conditions; so protected, the compound may be used as a reactant which has effectively only one reactive functional group. After the desired reaction (involving the other functional group) is complete, the protected group may be “deprotected” to return it to its original functionality.


For example, a hydroxy group may be protected as an ether (—OR) or an ester (—OC(═O)R), for example, as: a t-butyl ether; a benzyl, benzhydryl(diphenylmethyl), or trityl(triphenylmethyl)ether; a trimethylsilyl or t-butyldimethylsilyl ether; or an acetyl ester (—OC(═O)CH3, —OAc).


For example, an aldehyde or ketone group may be protected as an acetal (R—CH(OR)2) or ketal (R2C(OR)2), respectively, in which the carbonyl group (>C═O) is converted to a diether (>C(OR)2), by reaction with, for example, a primary alcohol. The aldehyde or ketone group is readily regenerated by hydrolysis using a large excess of water in the presence of acid.


For example, an amine group may be protected, for example, as an amide (—NRCO—R) or a urethane (—NRCO—OR), for example, as: a methyl amide (—NHCO—CH3); a benzyloxycarbonyl amide (—NHCO—OCH2C6H5, —NH-Cbz); as a t-butoxycarbonyl amine (—NHCO—OC(CH3)3, —NH-Boc); a 2-biphenyl-2-propoxycarbonyl amine (—NHCO—OC(CH3)2C6H4C6H5, —NH-Bpoc), as a 9-fluorenylmethoxycarbonyl amine (—NH-Fmoc), as a 6-nitroveratryloxycarbonyl amine (—NH-Nvoc), as a 2-trimethylsilylethyloxycarbonyl amine (—NH-Teoc), as a 2,2,2-trichloroethyloxycarbonyl amine (—NH-Troc), as an allyloxycarbonyl amine (—NH-Alloc), as a 2(-phenylsulfonyl)ethyloxycarbonyl amine (—NH-Psec); or, in suitable cases (e.g., cyclic amines), as a nitroxide radical (>N—O).


For example, a carboxylic acid group may be protected as an ester for example, as: an C1-7alkyl ester (e.g., a methyl ester; a t-butyl ester); a C1-7haloalkyl ester (e.g., a C1-7trihaloalkyl ester); a triC1-7alkylsilyl-C1-7alkyl ester; or a C5-20aryl-C1-7alkyl ester (e.g., a benzyl ester; a nitrobenzyl ester); or as an amide, for example, as a methyl amide.


For example, a thiol group may be protected as a thioether (—SR), for example, as: a benzyl thioether; an acetamidomethyl ether (—S—CH2NHC(═O)CH3).


Prodrugs

It may be convenient or desirable to prepare, purify, and/or handle the compound in the form of a prodrug. The term “prodrug,” as used herein, pertains to a compound which, when metabolised (e.g., in vivo), yields the desired active compound. Typically, the prodrug is inactive, or less active than the desired active compound, but may provide advantageous handling, administration, or metabolic properties.


For example, some prodrugs are esters of the active compound (e.g., a physiologically acceptable metabolically labile ester). During metabolism, the ester group (—C(═O)OR) is cleaved to yield the active drug. Such esters may be formed by esterification, for example, of any of the carboxylic acid groups (—C(═O)OH) in the parent compound, with, where appropriate, prior protection of any other reactive groups present in the parent compound, followed by deprotection if required.


Also, some prodrugs are activated enzymatically to yield the active compound, or a compound which, upon further chemical reaction, yields the active compound (for example, as in ADEPT, GDEPT, LIDEPT, etc.). For example, the prodrug may be a sugar derivative or other glycoside conjugate, or may be an amino acid ester derivative.


General Chemical Synthesis

Several methods for the chemical synthesis of IQ compounds are described herein. These and/or other well known methods may be modified and/or adapted in known ways in order to facilitate the synthesis of additional compounds described herein.


All reagents were either purchased from common commercial sources or synthesised in accordance with known literature procedures. Commercial reagents were used without further purification unless otherwise stated. Microwave reactions were conducted using a CEM Discover. Flash column chromatography was conducted using pre-packed silica Biotage® SNAP (KP-Sil) cartiridges. Ion exchange chromatography was performed using Isolute® Flash SCX-2 cartridges.


Abbreviations

APCI: atmospheric pressure chemical ionisation.


BBr3: boron tribromide.


BINAP: 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl.


Boc: tert-butyloxycarbonyl.


CH2Cl2: dichloromethane.


CV: column volume.


DEAD: diethylazodicarboxylate.


DIAD: diisopropyl azodicarboxylate.


DIPEA: N,N-diisopropylamine


DMA: dimethyl acetamide.


DMAP: 4-dimethylaminopyridine


DME: dimethoxyethane.


DMF: N,N-dimethylformamide.


Dppf: 1,1′-Bis(diphenylphosphino)ferrocene.


EDCl: 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide.


ES: electrospray.


EtOAc: ethyl acetate.


h: hour(s).


HATU: 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate.


IPA: isopropyl alcohol.


LDA: lithium diisopropylamide.


MCPBA: meta-Chloroperoxybenzoic acid


min: minute(s).


Ms/mesyl: methane sulfonyl


PFPA: perfluorophthalic anhydride.


PPh3: triphenyl phosphine.


PS: polymer supported.


Py: pyridine.


Rf: retention factor


Rt: retention time.


RT: room temperature.


SCX: strong cation exchange


SEM: 2-(trimethylsilyl)ethoxymethyl.


TBAF: tetra-n-butylammonium fluoride.


TBDMS: tert-butyldimethylsilyl.


TBDPS: tert-butyldiphenyllsilyl.


TBTU: O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate.


TFA: trifluoroacetic acid.


THF: tetrahydrofuran.


Ts/tosyl; 4-toluenesulfonyl.


The general synthetic methods for the synthesis of 2H-isoquinolin-1-ones 5 are illustrated below:


Route 1: Synthesis of 2H-isoquinolin-1-ones 5 via Cyclisation



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Acid 1 can be reacted with amine 2 (e.g., N,N-diethylamine) to yield amide 3, either by utilising standard amine coupling procedures (e.g., EDCI, HATU, etc.) or converting the acid 1 into the corresponding acid chloride (or mixed anhydride) and reacting with the amine 2 (see, e.g., Le et al., 2004). The 2-H-isoquinolin-1-one 5 can be prepared by in situ deprotonation of 2-methyl-benzamide derivative 3 with a suitable base (e.g., n-BuLi, sec-BuLi, t-BuLi, LDA, etc.) in THF (or similar suitable aprotic solvent) at −78° C., then reacting with the required nitrile 4 (see, e.g., Hattori et al., 2006).


Route 2: Synthesis of 2H-isoquinolin-1-ones 5 via Organopalladium Cross-Coupling



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The 2H-isoquinolin-1-one 5 can be synthesised by a palladium-mediated cross-coupling from the corresponding aryl halide 11 (e.g. chloride) and the corresponding boronic acid or ester (Suzuki cross-coupling).


Route 2a: Alternative synthesis of 2H-isoquinolin-1-ones 5 via Organopalladium Cross-Coupling



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In an alternative route, the 2H-isoquinolin-1-one 5 can be synthesised by a palladium-mediated cross-coupling from the corresponding aryl or heteroaryl halides 13 (e.g., bromide) and the corresponding boronic ester 35 (Suzuki cross-coupling). The boronic ester 35 can be accessed by a palladium-mediated cross-coupling from the corresponding 3-halo-2H-isoquinolin-1-one 11 (e.g., chloride) with a suitable diboron reagent (e.g. bis(pinacolato)diboron), and a suitable source of palladium (e.g., Pd(PPh3)4, PdCl2(PPh3)2) in an appropriate solvent (e.g., THF, DMF, DME, DCE, toluene, etc.).


For the Suzuki cross-coupling, 3-halo-2H-isoquinolin-1-one (e.g chloride) 11 can be reacted with a suitable boronic acid or ester 12 in the presence of a suitable base (e.g., K2CO3, NaOt-Bu, K3PO4, etc.), a suitable source of palladium (e.g., Pd(PPh3)4, PdCl2(PPh3)2, etc.) and a ligand (e.g., P(t-Bu)3, BINAP, etc.) in an appropriate solvent (e.g., THF, DME, DCE, toluene, etc.).


The 3-chloro-2H-isoquinolin-1-one 11 can be synthesised from indan-1,2-dione 2-oxime 10 (see, e.g., Merchant et al., 1984) via Beckmann rearrangement followed by treatment with PCl5.


Indan-1,2-dione 2-oxime 10 can be accessed from commercial sources or prepared from commercially available indanones 9 by nitrosation or from aldehyde 6 via chain extension, cyclisation and nitrosation (see, e.g., Musso et al., 2003).


The general synthetic methods for the synthesis of nitrile intermediates 4 and boronic acid or boronic ester intermediates 12 are illustrated below:


Synthesis of Aryl Nitrile 4 from Aryl Bromide 13



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The nitrile 4 can be accessed by a palladium-mediated cyanide insertion from the corresponding carboaryl or heteroaryl halide 13 (e.g., iodide, bromide, chloride) with a source of cyanide e.g., Zn(CN)2, Cu(CN)2, and a suitable source of palladium (e.g., Pd(PPh3)4, PdCl2(PPh3)2) in an appropriate solvent (e.g., THF, DMF, DME, DCE, toluene, etc.).


Synthesis of Boronic Acid or Boronic Ester Intermediate 12 from Aryl Halide 13



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The boronic acid or ester 12 can be accessed by a palladium-mediated cross-coupling from the corresponding aryl(heteroaryl) halide 13 (e.g., iodide, bromide, chloride) with bis(pinacolato)diboron, and a suitable source of palladium (e.g., Pd(PPh3)4, PdCl2(PPh3)2) in an appropriate solvent (e.g., THF, DMF, DME, DCE, toluene, etc.).


Synthesis of Amine 17 from Alkyl Bromide 15



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The amine 17 can be accessed by bromide displacement from the corresponding halide 15 (e.g., iodide, bromide, chloride) and an appropriate amine 16 in an appropriate solvent (e.g., THF, DMF, CH2Cl2 etc.).


This method is exemplified in Scheme 5 with benzyl or heteroarylmethyl bromides, but it is understood that the same approach can be extended to other examples of A-aryl-L3PR1-Br. The same method can be used for any amine 16 as defined in the claims, including aromatic heterocycles HNRC5RD5 (e.g., imidazole, pyrazole, etc.).


Synthesis of Amine 17 from Aldehyde 18



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The amine 17 can be accessed by standard reductive amination conditions from the corresponding aldehyde 18 and an appropriate amine 16 in an appropriate solvent (e.g., DCE etc.), with the use a standard reducing reagent (e.g., sodium triacetoxy borohydride, sodium borohydride, etc.).


Synthesis of Amide 20 from Acid 19



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The amide 20 can be accessed by standard amine coupling conditions from the corresponding acid (or acid chloride) 19 and an appropiate amine 16 in an appropriate solvent (e.g., THF, DMF, CH2Cl2 etc.), with a suitable base (e.g., DIPEA, Et3N etc.) with the use a standard amine coupling reagent (e.g., HATU, TBTU, EDCI etc.).


Alternatively, the amide 20 can be accessed by standard amine coupling conditions from the corresponding acid chloride 19 and an appropiate amine 16 in an appropriate solvent (e.g., THF, DMF, CH2Cl2 etc.), with a suitable base (e.g., DIPEA, Et3N etc.).


Synthesis of Amide 20 from Acid 19



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The same method from Scheme 7 can be applied using a carboxylic acid (or acid chloride) 36, with an amine 16, to afford amide 37.


Synthesis of Sulfonamide 22 from Sulfonyl Chloride 21



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The sulfonamide 22 can be prepared from the corresponding sulfonyl chloride 21 and an appropriate amine 16 in an appropriate solvent (e.g., THF, CH2Cl2 etc.), with a suitable base (e.g., DIPEA, Et3N etc.).


Synthesis of Amino-Heteroaryl Nitrile 24 from Halo-Heteroaryl Nitrile 23



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Halo-heteroaryl 23 can be reacted with amine 16 to yield amino-heteroaryl 24 (see, e.g., Nettekoven et al., 2006) either by heating in acetonitrile (or other suitable solvent) or by irradiation using microwave heating in acetonitrile (or other suitable solvent).


The general synthetic methods for the synthesis of 2H-isoquinolin-1-ones 5 are illustrated below:


Synthesis of 2H-isoquinolin-1-ones 5 via Organometal Cross-Coupling



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The 2H-isoquinolin-1-one 5 can be synthesised by palladium-mediated cross-coupling of an aryl halide 25 and a suitable trialkylaluminium reagent 26 (see, e.g., Molander et al., 2003) in the presence of a suitable source of palladium (e.g., Pd(PPh3)4, PdCl2(PPh3)2, etc.) and CeCl3 in an appropriate solvent (e.g., THF, DME, DCE, toluene, dioxane, etc.).


The aryl halide 25 can alternatively be reacted with a suitable organo-zinc halide or diorgano-zinc compound 27 (see, e.g., Hughes et al., 2007) in the presence of a suitable source of palladium (e.g., Pd(PPh3)4, PdCl2(PPh3)2, etc.), a ligand (e.g., P(t-Bu)3, BINAP, etc.) in an appropriate solvent (e.g., THF, DME, DCE, toluene, dioxane, etc.).


Synthesis of 2H-isoquinolin-1-ones 5 via Sonogashira Coupling



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The 2H-isoquinolin-1-one 5 can be synthesised by palladium/copper-mediated cross-coupling (Sonogashira coupling) of an aryl halide 25 and a suitable alkynyl reagent 38 in the presence of a base (e.g., DIPEA, triethylamine, pyrrolidine, piperidine, Cs2CO3, etc.), a suitable source of palladium (e.g., Pd(PPh3)4, PdCl2(PPh3)2, etc.) and a ligand (e.g., PPh3, P(t-Bu)3, etc.) in an appropriate solvent (e.g., THF, DMF, DME, DCE, toluene, dioxane, etc.).


Synthesis of 2H-Isoquinolin-1-Ones 30 Via N-Acylation



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The amide 30 can be accessed by standard amine coupling conditions from the corresponding amine 28 and an appropiate acid 29 in an appropriate solvent (e.g., THF, DMF, CH2Cl2 etc.), with a suitable base (e.g., DIPEA, Et3N etc.) with the use a standard amine coupling reagent (e.g., HATU, TBTU, EDCI etc.).


Alternatively, the amide 30 can be accessed by standard amine coupling conditions from the corresponding amine 28 and an appropiate acid chloride 29 in an appropriate solvent (e.g., THF, DMF, CH2Cl2 etc.), with a suitable base (e.g., DIPEA, Et3N etc.).


Synthesis of 2H-Isoquinolin-1-Ones 32 Via Urea Formation



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The urea 32 can be accessed by standard urea formation conditions from the corresponding amine 28 and an appropiate isocyanate 31 in an appropriate solvent (e.g, DMF, CH2Cl2 etc.).


Synthesis of 2H-Isoquinolin-1-Ones 34 Via N-Acylation



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The amide 34 can be accessed by standard amine coupling conditions from the corresponding acid 33 and an appropiate amine 16 in an appropriate solvent (e.g., THF, DMF, CH2Cl2 etc.), with a suitable base (e.g., DIPEA, Et3N etc.) with the use a standard amine coupling reagent (e.g., HATU, TBTU, EDCI etc.).


Synthesis of 2H-Isoquinolin-1-Ones 40 Via N-Acylation



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The same method from Scheme 13 can be applied using a carboxylic acid 39, with an amine 16, to afford amide 40.


Compositions

One aspect of the present invention pertains to a composition (e.g., a pharmaceutical composition) comprising an IQ compound, as described herein, and a pharmaceutically acceptable carrier, diluent, or excipient.


Another aspect of the present invention pertains to a method of preparing a composition (e.g., a pharmaceutical composition) comprising mixing an IQ compound, as described herein, and a pharmaceutically acceptable carrier, diluent, or excipient.


Uses

The IQ compounds, as described herein, are useful, for example, in the treatment of disorders (e.g., diseases) that are ameliorated by the inhibition of PARP (e.g., PARP1, TNKS1, TNKS2, etc.) and/or the inhibition of Wnt signalling, as described herein.


Use in Methods of Inhibiting PARP (e.g., PARP1, TNKS1, TNKS2, etc.)

One aspect of the present invention pertains to a method of inhibiting PARP (e.g., PARP1, TNKS1, TNKS2, etc.) in a cell, in vitro or in vivo, comprising contacting the cell with an effective amount of an IQ compound, as described herein.


One aspect of the present invention pertains to a method of inhibiting TNKS1 and/or TNKS2 in a cell, in vitro or in vivo, comprising contacting the cell with an effective amount of an IQ compound, as described herein.


One of ordinary skill in the art is readily able to determine whether or not a candidate compound inhibits PARP (e.g., PARP1, TNKS1, TNKS2, etc.). For example, suitable assays are described herein or are known in the art.


In one embodiment, the method is performed in vitro.


In one embodiment, the method is performed in vivo.


In one embodiment, the IQ compound is provided in the form of a pharmaceutically acceptable composition.


Any type of cell may be treated, including but not limited to, adipose, lung, gastrointestinal (including, e.g., bowel, colon), breast (mammary), ovarian, prostate, liver (hepatic), kidney (renal), bladder, pancreas, brain, and skin.


For example, a sample of cells may be grown in vitro and a compound brought into contact with said cells, and the effect of the compound on those cells observed. As an example of “effect,” the morphological status of the cells (e.g., alive or dead, etc.) may be determined. Where the compound is found to exert an influence on the cells, this may be used as a prognostic or diagnostic marker of the efficacy of the compound in methods of treating a patient carrying cells of the same cellular type.


Use in Methods of Inhibiting Wnt Signalling

One aspect of the present invention pertains to a method of inhibiting Wnt signalling in a cell, in vitro or in vivo, comprising contacting the cell with an effective amount of an IQ compound, as described herein.


One of ordinary skill in the art is readily able to determine whether or not a candidate compound inhibits Wnt signalling. For example, suitable assays are described herein or are known in the art.


In one embodiment, the method is performed in vitro.


In one embodiment, the method is performed in vivo.


In one embodiment, the IQ compound is provided in the form of a pharmaceutically acceptable composition.


Use in Methods of Inhibiting Cell Proliferation, etc.

The IQ compounds described herein, e.g., (a) regulate (e.g., inhibit) cell proliferation; (b) inhibit cell cycle progression; (c) promote apoptosis; or (d) a combination of one or more of these.


One aspect of the present invention pertains to a method of regulating (e.g., inhibiting) cell proliferation (e.g., proliferation of a cell), inhibiting cell cycle progression, promoting apoptosis, or a combination of one or more these, in vitro or in vivo, comprising contacting a cell with an effective amount of an IQ compound, as described herein.


In one embodiment, the method is a method of regulating (e.g., inhibiting) cell proliferation (e.g., proliferation of a cell), in vitro or in vivo, comprising contacting a cell with an effective amount of an IQ compound, as described herein.


In one embodiment, the method is performed in vitro.


In one embodiment, the method is performed in vivo.


In one embodiment, the IQ compound is provided in the form of a pharmaceutically acceptable composition.


Any type of cell may be treated, including but not limited to, lung, gastrointestinal (including, e.g., bowel, colon), breast (mammary), ovarian, prostate, liver (hepatic), kidney (renal), bladder, pancreas, brain, and skin.


One of ordinary skill in the art is readily able to determine whether or not a candidate compound regulates (e.g., inhibits) cell proliferation, etc. For example, assays which may conveniently be used to assess the activity offered by a particular compound are described herein.


For example, a sample of cells (e.g., from a tumour) may be grown in vitro and a compound brought into contact with said cells, and the effect of the compound on those cells observed. As an example of “effect,” the morphological status of the cells (e.g., alive or dead, etc.) may be determined. Where the compound is found to exert an influence on the cells, this may be used as a prognostic or diagnostic marker of the efficacy of the compound in methods of treating a patient carrying cells of the same cellular type.


Use in Methods of Therapy

Another aspect of the present invention pertains to an IQ compound, as described herein, for use in a method of treatment of the human or animal body by therapy.


Use in the Manufacture of Medicaments

Another aspect of the present invention pertains to use of an IQ compound, as described herein, in the manufacture of a medicament for use in treatment.


In one embodiment, the medicament comprises the IQ compound.


Methods of Treatment

Another aspect of the present invention pertains to a method of treatment comprising administering to a patient in need of treatment a therapeutically effective amount of an IQ compound, as described herein, preferably in the form of a pharmaceutical composition.


Disorders Treated—Disorders Ameliorated by the Inhibition of PARP (e.g., PARP1, TNKS1, TNKS2, etc.)

In one embodiment (e.g., of use in methods of therapy, of use in the manufacture of medicaments, of methods of treatment), the treatment is treatment of a disorder (e.g., a disease) that is ameliorated by the inhibition of PARP.


In one embodiment (e.g., of use in methods of therapy, of use in the manufacture of medicaments, of methods of treatment), the treatment is treatment of a disorder (e.g., a disease) that is ameliorated by the inhibition of TNKS1 and/or TNKS2.


Disorders Treated—Proliferative Conditions

In one embodiment (e.g., of use in methods of therapy, of use in the manufacture of medicaments, of methods of treatment), the treatment is treatment of: a proliferative condition.


The term “proliferative condition,” as used herein, pertains to an unwanted or uncontrolled cellular proliferation of excessive or abnormal cells which is undesired, such as neoplastic or hyperplastic growth.


In one embodiment, the treatment is treatment of: a proliferative condition characterised by benign, pre-malignant, or malignant cellular proliferation, including for example: neoplasms, hyperplasias, and tumours (e.g., histocytoma, glioma, astrocyoma, osteoma), cancers (see below), psoriasis, bone diseases, fibroproliferative disorders (e.g., of connective tissues), pulmonary fibrosis, atherosclerosis, smooth muscle cell proliferation in the blood vessels, such as stenosis or restenosis following angioplasty.


Disorders Treated—Cancer

In one embodiment (e.g., of use in methods of therapy, of use in the manufacture of medicaments, of methods of treatment), the treatment is treatment of cancer.


In one embodiment, the treatment is treatment of cancer characterised by, or further characterised by, cancer cells which overexpress PARP.


In one embodiment, the treatment is treatment of cancer characterised by, or further characterised by, cancer cells which overexpress TNKS1 and/or TNKS2.


In one embodiment, the treatment is treatment of lung cancer, small cell lung cancer, non-small cell lung cancer, gastrointestinal cancer, stomach cancer, bowel cancer, colon cancer, rectal cancer, colorectal cancer, thyroid cancer, breast cancer, ovarian cancer, endometrial cancer, prostate cancer, testicular cancer, liver cancer, kidney cancer, renal cell carcinoma, bladder cancer, pancreatic cancer, brain cancer, glioma, sarcoma, osteosarcoma, bone cancer, nasopharyngeal cancer (e.g., head cancer, neck cancer), skin cancer, squamous cancer, Kaposi's sarcoma, melanoma, malignant melanoma, lymphoma, or leukemia.


In one embodiment, the treatment is treatment of:

    • a carcinoma, for example a carcinoma of the bladder, breast, colon (e.g., colorectal carcinomas such as colon adenocarcinoma and colon adenoma), kidney, epidermal, liver, lung (e.g., adenocarcinoma, small cell lung cancer and non-small cell lung carcinomas), oesophagus, gall bladder, ovary, pancreas (e.g., exocrine pancreatic carcinoma), stomach, cervix, thyroid, prostate, skin (e.g., squamous cell carcinoma);
    • a hematopoietic tumour of lymphoid lineage, for example leukemia, acute lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, or Burkett's lymphoma;
    • a hematopoietic tumour of myeloid lineage, for example acute and chronic myelogenous leukemias, myelodysplastic syndrome, or promyelocytic leukemia;
    • a tumour of mesenchymal origin, for example fibrosarcoma or habdomyosarcoma;
    • a tumour of the central or peripheral nervous system, for example astrocytoma, neuroblastoma, glioma or schwannoma;
    • melanoma; seminoma; teratocarcinoma; osteosarcoma; xenoderoma pigmentoum; keratoctanthoma; thyroid follicular cancer; or Kaposi's sarcoma.


In one embodiment, the treatment is treatment of solid tumour cancer.


In one embodiment, the treatment is treatment of cancer head and neck cancer; nervous system cancer; lung/mediastinum cancer; breast cancer; oesophagus cancer; stomach cancer; liver cancer; biliary tract cancer; pancreatic cancer; small bowel cancer; large bowel cancer; gynaecological cancer; genito-urinary cancer; thyroid gland cancer; adrenal gland cancer; skin cancer; bone sarcoma; soft tissue sarcoma; paediatric malignancy; Hodgkin's disease; non-Hodgkin's lymphoma; myeloma; leukaemia; or metastasis from an unknown primary site.


In one embodiment, the treatment is treatment of cancer metastasis.


In one embodiment, the cancer is characterised by, or further characterised by, cancer stem cells.


The anti-cancer effect may arise through one or more mechanisms, including but not limited to, the regulation of cell proliferation, the inhibition of cell cycle progression, the inhibition of angiogenesis (the formation of new blood vessels), the inhibition of metastasis (the spread of a tumour from its origin), the inhibition of cell migration (the spread of cancer cells to other parts of the body), the inhibition of invasion (the spread of tumour cells into neighbouring normal structures), or the promotion of apoptosis (programmed cell death). The compounds of the present invention may be used in the treatment of the cancers described herein, independent of the mechanisms discussed herein.


Disorders Treated—Non-Cancer Indications Related to Tankyrase Inhibition

In one embodiment, the treatment is treatment of: a neurodegenerative disorder, such as multiple sclerosis (MS); a neurological disorder associated with demyelination; neonatal hypoxic ischemic encephalopathy (HIE); neonatal periventricular leukomalacia (PVL); a cardiac related pathology, such as myocardial infarction; cardiac damage (e.g., to repair cardiac damage); an infectious disease, such as a pathology related to Herpes Simplex Virus (HSV); a pathology related to Epstein-Barr Virus (EBV); a metabolic disease, such as a metabolic disease where glucose uptake is dysfunctional, such as diabetes, such as type 2 diabetes; or fibrosis (e.g., lung fibrosis).


In one embodiment, the treatment is treatment of: a neurodegenerative disorder, such as multiple sclerosis (MS); neonatal hypoxic ischemic encephalopathy (HIE); neonatal periventricular leukomalacia (PVL); a cardiac related pathology, such as myocardial infarction; a pathology related to Herpes Simplex Virus (HSV); a pathology related to Epstein-Barr Virus (EBV); or a metabolic disease such as type 2 diabetes.


Disorder Treated—Non-Cancer Indications Related to Wnt Signalling

In one embodiment, the treatment is treatment of: Alzheimer's disease; late onset Alzheimer's disease; Dupuytren skin disease; tooth agenesis; vascular defects in the eye; Osteoperosis-pseudoglioma Syndrome (OPPG); exudative vitreoretinopathy; familial exudative vitreoretinopathy; retinal angiogenesis; schizophrenia; osteoporosis; dermal hypoplasia; XX sex reversal; Mullerian-duct regression and virilization; SERKAL syndrome; anonychia; hyponychia; sclerosteosis; van Buchem disease; Fuhrmann syndrome; odonto-onchyo-dermal hypoplasia; Type 2 diabetes; obesity; early onset obesity; a nephropathy, such as HIV-associated nephropathy; early coronary disease; bone density defects; tetra-amelia syndrome; split-hand/foot malformation; caudal duplication; Fuhrmann syndrome; odonto-onycho-dermal dysplasia; skeletal dysplasia; focal dermal hypoplasia; autosomal recessive anonychia; or neural tube defects.


In one embodiment, the treatment is treatment of: Alzheimer's disease; Dupuytren skin disease; tooth agenesis; exudative vitreoretinopathy; schizophrenia; osteoporosis; dermal hypoplasia; XX sex reversal; anonychia; hyponychia; sclerosteosis; van Buchem disease; Fuhrmann syndrome; odonto-onchyo-dermal hypoplasia; early onset obesity; or a nephropathy, such as HIV-associated nephropathy.


Treatment

The term “treatment,” as used herein in the context of treating a disorder, pertains generally to treatment of a human or an animal (e.g., in veterinary applications), in which some desired therapeutic effect is achieved, for example, the inhibition of the progress of the disorder, and includes a reduction in the rate of progress, a halt in the rate of progress, alleviation of symptoms of the disorder, amelioration of the disorder, and cure of the disorder. Treatment as a prophylactic measure (i.e., prophylaxis) is also included. For example, use with patients who have not yet developed the disorder, but who are at risk of developing the disorder, is encompassed by the term “treatment.”


For example, treatment includes the prophylaxis of cancer, reducing the incidence of cancer, alleviating the symptoms of cancer, etc.


The term “therapeutically-effective amount,” as used herein, pertains to that amount of a compound, or a material, composition or dosage form comprising a compound, which is effective for producing some desired therapeutic effect, commensurate with a reasonable benefit/risk ratio, when administered in accordance with a desired treatment regimen.


Combination Therapies

The term “treatment” includes combination treatments and therapies, in which two or more treatments or therapies are combined, for example, sequentially or simultaneously. For example, the compounds described herein may also be used in combination therapies, e.g., in conjunction with other agents. Examples of treatments and therapies include, but are not limited to, chemotherapy (the administration of active agents, including, e.g., drugs, antibodies (e.g., as in immunotherapy), prodrugs (e.g., as in photodynamic therapy, GDEPT, ADEPT, etc.); surgery; radiation therapy; photodynamic therapy; gene therapy; and controlled diets.


One aspect of the present invention pertains to a compound as described herein, in combination with one or more (e.g., 1, 2, 3, 4, etc.) additional therapeutic agents, as described below.


The particular combination would be at the discretion of the physician who would select dosages using his common general knowledge and dosing regimens known to a skilled practitioner.


The agents (i.e., the compound described herein, plus one or more other agents) may be administered simultaneously or sequentially, and may be administered in individually varying dose schedules and via different routes. For example, when administered sequentially, the agents can be administered at closely spaced intervals (e.g., over a period of 5-10 minutes) or at longer intervals (e.g., 1, 2, 3, 4 or more hours apart, or even longer periods apart where required), the precise dosage regimen being commensurate with the properties of the therapeutic agent(s).


The agents (i.e., the compound described here, plus one or more other agents) may be formulated together in a single dosage form, or alternatively, the individual agents may be formulated separately and presented together in the form of a kit, optionally with instructions for their use.


Examples of additional agents/therapies that may be co-administered/combined with treatment with the IQ compounds described herein include the following: antimetabolites; alkylating agents; spindle poisons; topoisomerase inhibitors; DNA binding agents; kinase inhibitors; therapeutic antibodies; PARP inhibitors; NAD metabolism inhibitors; metabolic inhibitors; targeted agents; endocrine agents; etc.


Other Uses

The IQ compounds described herein may also be used as cell culture additives to inhibit PARP (e.g., PARP1, TNKS1, TNKS2, etc.), to inhibit Wnt signalling, etc.


The IQ compounds described herein may also be used as part of an in vitro assay, for example, in order to determine whether a candidate host is likely to benefit from treatment with the compound in question.


The IQ compounds described herein may also be used as a standard, for example, in an assay, in order to identify other active compounds, other PARP (e.g., PARP1, TNKS1, TNKS2, etc.) inhibitors, other Wnt signalling inhibitors, etc.


Kits

One aspect of the invention pertains to a kit comprising (a) an IQ compound as described herein, or a composition comprising an IQ compound as described herein, e.g., preferably provided in a suitable container and/or with suitable packaging; and (b) instructions for use, e.g., written instructions on how to administer the compound or composition.


The written instructions may also include a list of indications for which the active ingredient is a suitable treatment.


Routes of Administration

The IQ compound or pharmaceutical composition comprising the IQ compound may be administered to a subject by any convenient route of administration, whether systemically/peripherally or topically (i.e., at the site of desired action).


Routes of administration include, but are not limited to, oral (e.g., by ingestion); buccal; sublingual; transdermal (including, e.g., by a patch, plaster, etc.); transmucosal (including, e.g., by a patch, plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by eyedrops); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., via an aerosol, e.g., through the mouth or nose); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal; by implant of a depot or reservoir, for example, subcutaneously or intramuscularly.


The Subject/Patient

The subject/patient may be a chordate, a vertebrate, a mammal, a placental mammal, a marsupial (e.g., kangaroo, wombat), a rodent (e.g., a guinea pig, a hamster, a rat, a mouse), murine (e.g., a mouse), a lagomorph (e.g., a rabbit), avian (e.g., a bird), canine (e.g., a dog), feline (e.g., a cat), equine (e.g., a horse), porcine (e.g., a pig), ovine (e.g., a sheep), bovine (e.g., a cow), a primate, simian (e.g., a monkey or ape), a monkey (e.g., marmoset, baboon), an ape (e.g., gorilla, chimpanzee, orangutang, gibbon), or a human.


Furthermore, the subject/patient may be any of its forms of development, for example, a foetus.


In one preferred embodiment, the subject/patient is a human.


Formulations

While it is possible for an IQ compound to be administered alone, it is preferable to present it as a pharmaceutical formulation (e.g., composition, preparation, medicament) comprising at least one IQ compound, as described herein, together with one or more other pharmaceutically acceptable ingredients well known to those skilled in the art, including, but not limited to, pharmaceutically acceptable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, anti-oxidants, lubricants, stabilisers, solubilisers, surfactants (e.g., wetting agents), masking agents, colouring agents, flavouring agents, and sweetening agents. The formulation may further comprise other active agents, for example, other therapeutic or prophylactic agents.


Thus, the present invention further provides pharmaceutical compositions, as defined above, and methods of making a pharmaceutical composition comprising mixing at least one IQ compound, as described herein, together with one or more other pharmaceutically acceptable ingredients well known to those skilled in the art, e.g., carriers, diluents, excipients, etc. If formulated as discrete units (e.g., tablets, etc.), each unit contains a predetermined amount (dosage) of the compound.


The term “pharmaceutically acceptable,” as used herein, pertains to compounds, ingredients, materials, compositions, dosage forms, etc., which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of the subject in question (e.g., human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each carrier, diluent, excipient, etc. must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation.


Suitable carriers, diluents, excipients, etc. can be found in standard pharmaceutical texts, for example, Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients, 5th edition, 2005.


The formulations may be prepared by any methods well known in the art of pharmacy. Such methods include the step of bringing into association the compound with a carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the compound with carriers (e.g., liquid carriers, finely divided solid carrier, etc.), and then shaping the product, if necessary.


The formulation may be prepared to provide for rapid or slow release; immediate, delayed, timed, or sustained release; or a combination thereof.


Formulations may suitably be in the form of liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), elixirs, syrups, electuaries, mouthwashes, drops, tablets (including, e.g., coated tablets), granules, powders, losenges, pastilles, capsules (including, e.g., hard and soft gelatin capsules), cachets, pills, ampoules, boluses, suppositories, pessaries, tinctures, gels, pastes, ointments, creams, lotions, oils, foams, sprays, mists, or aerosols.


Formulations may suitably be provided as a patch, adhesive plaster, bandage, dressing, or the like which is impregnated with one or more compounds and optionally one or more other pharmaceutically acceptable ingredients, including, for example, penetration, permeation, and absorption enhancers. Formulations may also suitably be provided in the form of a depot or reservoir.


The compound may be dissolved in, suspended in, or mixed with one or more other pharmaceutically acceptable ingredients. The compound may be presented in a liposome or other microparticulate which is designed to target the compound, for example, to blood components or one or more organs.


Formulations suitable for oral administration (e.g., by ingestion) include liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), elixirs, syrups, electuaries, tablets, granules, powders, capsules, cachets, pills, ampoules, boluses.


Formulations suitable for buccal administration include mouthwashes, losenges, pastilles, as well as patches, adhesive plasters, depots, and reservoirs. Losenges typically comprise the compound in a flavored basis, usually sucrose and acacia or tragacanth. Pastilles typically comprise the compound in an inert matrix, such as gelatin and glycerin, or sucrose and acacia. Mouthwashes typically comprise the compound in a suitable liquid carrier.


Formulations suitable for sublingual administration include tablets, losenges, pastilles, capsules, and pills.


Formulations suitable for oral transmucosal administration include liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), mouthwashes, losenges, pastilles, as well as patches, adhesive plasters, depots, and reservoirs.


Formulations suitable for non-oral transmucosal administration include liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), suppositories, pessaries, gels, pastes, ointments, creams, lotions, oils, as well as patches, adhesive plasters, depots, and reservoirs.


Formulations suitable for transdermal administration include gels, pastes, ointments, creams, lotions, and oils, as well as patches, adhesive plasters, bandages, dressings, depots, and reservoirs.


Tablets may be made by conventional means, e.g., compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the compound in a free-flowing form such as a powder or granules, optionally mixed with one or more binders (e.g., povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropylmethyl cellulose); fillers or diluents (e.g., lactose, microcrystalline cellulose, calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, silica); disintegrants (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose); surface-active or dispersing or wetting agents (e.g., sodium lauryl sulfate); preservatives (e.g., methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid); flavours, flavour enhancing agents, and sweeteners. Moulded tablets may be made by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the compound therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with a coating, for example, to affect release, for example an enteric coating, to provide release in parts of the gut other than the stomach.


Ointments are typically prepared from the compound and a paraffinic or a water-miscible ointment base.


Creams are typically prepared from the compound and an oil-in-water cream base. If desired, the aqueous phase of the cream base may include, for example, at least about 30% w/w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof. The topical formulations may desirably include a compound which enhances absorption or penetration of the compound through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethylsulfoxide and related analogues.


Emulsions are typically prepared from the compound and an oily phase, which may optionally comprise merely an emulsifier (otherwise known as an emulgent), or it may comprise a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabiliser. It is also preferred to include both an oil and a fat. Together, the emulsifier(s) with or without stabiliser(s) make up the so-called emulsifying wax, and the wax together with the oil and/or fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.


Suitable emulgents and emulsion stabilisers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate. The choice of suitable oils or fats for the formulation is based on achieving the desired cosmetic properties, since the solubility of the compound in most oils likely to be used in pharmaceutical emulsion formulations may be very low. Thus the cream should preferably be a non-greasy, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic alkyl esters such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be used.


Formulations suitable for intranasal administration, where the carrier is a liquid, include, for example, nasal spray, nasal drops, or by aerosol administration by nebuliser, include aqueous or oily solutions of the compound.


Formulations suitable for intranasal administration, where the carrier is a solid, include, for example, those presented as a coarse powder having a particle size, for example, in the range of about 20 to about 500 microns which is administered in the manner in which snuff is taken, i.e., by rapid inhalation through the nasal passage from a container of the powder held close up to the nose.


Formulations suitable for pulmonary administration (e.g., by inhalation or insufflation therapy) include those presented as an aerosol spray from a pressurised pack, with the use of a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichoro-tetrafluoroethane, carbon dioxide, or other suitable gases.


Formulations suitable for ocular administration include eye drops wherein the compound is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the compound.


Formulations suitable for rectal administration may be presented as a suppository with a suitable base comprising, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols, for example, cocoa butter or a salicylate; or as a solution or suspension for treatment by enema.


Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the compound, such carriers as are known in the art to be appropriate.


Formulations suitable for parenteral administration (e.g., by injection), include aqueous or non-aqueous, isotonic, pyrogen-free, sterile liquids (e.g., solutions, suspensions), in which the compound is dissolved, suspended, or otherwise provided (e.g., in a liposome or other microparticulate). Such liquids may additionally contain other pharmaceutically acceptable ingredients, such as anti-oxidants, buffers, preservatives, stabilisers, bacteriostats, suspending agents, thickening agents, and solutes which render the formulation isotonic with the blood (or other relevant bodily fluid) of the intended recipient. Examples of excipients include, for example, water, alcohols, polyols, glycerol, vegetable oils, and the like. Examples of suitable isotonic carriers for use in such formulations include Sodium Chloride Injection, Ringer's Solution, or Lactated Ringer's Injection. Typically, the concentration of the compound in the liquid is from about 1 ng/mL to about 10 μg/mL, for example from about 10 ng/mL to about 1 μg/mL. The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.


Dosage

It will be appreciated by one of skill in the art that appropriate dosages of the IQ compounds, and compositions comprising the IQ compounds, can vary from patient to patient. Determining the optimal dosage will generally involve the balancing of the level of therapeutic benefit against any risk or deleterious side effects. The selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular IQ compound, the route of administration, the time of administration, the rate of excretion of the IQ compound, the duration of the treatment, other drugs, compounds, and/or materials used in combination, the severity of the disorder, and the species, sex, age, weight, condition, general health, and prior medical history of the patient. The amount of IQ compound and route of administration will ultimately be at the discretion of the physician, veterinarian, or clinician, although generally the dosage will be selected to achieve local concentrations at the site of action which achieve the desired effect without causing substantial harmful or deleterious side-effects.


Administration can be effected in one dose, continuously or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Methods of determining the most effective means and dosage of administration are well known to those of skill in the art and will vary with the formulation used for therapy, the purpose of the therapy, the target cell(s) being treated, and the subject being treated. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician, veterinarian, or clinician.


In general, a suitable dose of the IQ compound is in the range of about 10 μg to about 250 mg (more typically about 100 μg to about 25 mg) per kilogram body weight of the subject per day. Where the compound is a salt, an ester, an amide, a prodrug, or the like, the amount administered is calculated on the basis of the parent compound and so the actual weight to be used is increased proportionately.


EXAMPLES
Chemical Synthesis

The following examples are provided solely to illustrate the present invention and are not intended to limit the scope of the invention, as described herein.


Analytical Methods

Reverse Phase Preparative HPLC-MS: Mass-directed purification by preparative LC-MS using a preparative C-18 column (Phenomenex Luna C18 (2), 100×21.2 mm, 5 μm).


Analysis of products and intermediates has been carried out using reverse phase analytical HPLC-MS using the parameters set out below.


HPLC Analytical Methods:

AnalpH2_MeOH—4 min: Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water+0.1% formic acid; B=MeOH; 45° C.; % B: 0 min 5%, 1 min 37.5%, 3 min 95%, 3.51 min 5%, 4.5 min 5%; 2.25 mL/min.


AnalpH2_MeOH—4 min(1): Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 45° C.; % B: 0 min 5%, 1 min 37.5%, 3 min 95%, 3.51 min 5%, 4.5 min 5%; 2.25 mL/min.


AnalpH2_MeOH—4 min(2): Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 40° C.; % B: 0 min 5%, 1 min 37.5%, 3 min 95%, 3.51 min 5%, 4.5 min 5%; 2.25 mL/min.


AnalpH2_MeOH—4 min(3): Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 45° C.; % B: 0 min 5%, 1 min 37.5%, 3 min 95%, 3.51 min 5%, 4.0 min 5%; 2.25 mL/min.


AnalpH9_MeOH—4 min: Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water pH9 (Ammonium Bicarbonate 10 mM); B=MeOH; 45° C.; % B: 0 min 5%, 1 min 37.5%, 3 min 95%, 3.51 min 5%, 4.5 min 5%; 2.25 mL/min.


AnalpH9_MeOH—4 min(1): Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water pH9 (Ammonium Bicarbonate 10 mM); B=MeOH+0.1% formic acid; 45° C.; % B: 0 min 5%, 1 min 37.5%, 3 min 95%, 3.51 min 5%, 4.5 min 5%; 2.25 mL/min.


AnalpH9_MeOH—4 min(2): Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water pH9 (Ammonium Bicarbonate 10 mM); B=MeOH; 45° C.; % B: 0 min 5%, 1 min 37.5%, 3 min 95%, 3.51 min 5%, 4.0 min 5%; 2.25 mL/min.


AnalpH2_MeCN_TFA—4 min: Acquity UPLC BEH C18 1.7 μm, 50×2.1 mm; A=water+0.025% TFA; B=Acetonitrile+0.025% TFA; % B: 0 min 15%, 3 min 95%, 4 min 95%, 4.1 min 15%; 0.4 mL/min.


AnalpH2_MeCN_TFA—4 min(1): Acquity UPLC BEH C18 1.7 μm, 50×2.1 mm; A=water+0.025% TFA; B=Acetonitrile+0.025% TFA; % B: 0 min 50%, 4 min 80%, 6 min 80%, 6.1 min 50%; 0.3 mL/min.


AnalpH2_MeCN_FA—7 min(XTERRA1.m): Xterra C18 2.5 μm, 50×4.6 mm; A=water+0.1% FA; B=Acetonitrile+0.1% FA; % B: 0 min 20%, 4 min 90%, 7 min 90%, 7.1 min 20%; 1.0 mL/min.


AnalpH9_MeCN_AB—10 min (Develosil): Develosil C18 2.7 μm, 150×4.6 mm; A=water+0.01 M Ammonium bicarbonate; B=Acetonitrile; % B: 0 min 50%, 4 min 90%, 10 min 90%, 10.1 min 50%; 1.0 mL/min.


AnalpH2_MeOH_QC: Phenomenex Luna C18 (2) 3 μm, 150×4.6 mm; A=water+0.1% formic acid; B=MeOH; 35° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH2_MeOH_QC(1): Phenomenex Luna C18 (2) 3 μm, 150×4.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 40° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH2_MeOH_QC(2): Phenomenex Gemini C18 5 μm, 150×4.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 40° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH2_MeOH_QC(3): Phenomenex Gemini C18 5 μm, 250×4.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 40° C.; % B: 0 min 5%, 16 min 95%, 18 min 95%, 18.10 min 5%, 24.0 min 5%; 1.5 mL/min.


AnalpH9_MeOH_QC: Phenomenex Luna C18 (2)) 3 μm, 50×4.6 mm; A=water+pH9 (Ammonium Bicarbonate 10 mM); B=MeOH; 35° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH9_MeOH_QC(1): Phenomenex Luna C18 (2) 3 μm, 50×4.6 mm; A=water+pH9 (Ammonium Bicarbonate 10 mM); B=MeOH+0.1% formic acid; 35° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH2_MeOH_QC(Sunfire): Waters Sunfire C18 (2) 5 μm, 100×4.6 mm; A=water+0.1% formic acid; B=MeOH; 35° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH2_MeOH_QC(Sunfire1): Waters Sunfire C18 (2) 5 μm, 100×4.6 mm; A=water+0.1% formic acid; B=MeOH+0.1% formic acid; 40° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH9_MeOH_QC(Sunfire): Waters Sunfire C18 (2) 5 μm, 100×4.6 mm; A=water+pH9 (Ammonium Bicarbonate 10 mM); B=MeOH; 35° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


AnalpH9_MeOH_QC(Sunfire1): Waters Sunfire C18 (2) 5 μm, 100×4.6 mm; A=water+pH9 (Ammonium Bicarbonate 10 mM); B=MeOH+0.1% formic acid; 35° C.; % B: 0 min 5%, 7.5 min 95%, 10 min 95%, 10.10 min 5%, 13.0 min 5%; 1.5 mL/min.


Chiral HPLC Preparative Methods:

Chiral_Method—1: Daicel IA, 10 μm, 250×20 mm; MeOH+0.2% diethylamine


Chiral_Method—2: Daicel IA, 10 μm, 250×20 mm; 50% (MeCN+3% diethylamine)+50% EtOH


Chiral_Method—3: Daicel IA, 10 μm, 250×20 mm; EtOH+0.05% diethylamine


Synthesis of 2H-isoquinolin-1-ones of Formula 4-6



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Scheme A, Step A: Synthesis of N,N-Diethyl-benzamide Derivatives 2
N,N-Diethyl-2,3-dimethyl-benzamide



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To a stirred solution of 2,3-dimethyl-benzoic acid (1.52 g, 10.1 mmol) in CH2Cl2/DMF (118 mL/12 mL) was added N,N-diisopropylethylamine (1.76 mL, 10.1 mmol) and TBTU (3.25 g, 10.1 mmol) and the reaction mixture stirred at RT for 50 min. N,N-diethylamine (1.58 mL, 15.2 mmol) was added and the reaction mixture stirred for 18 h. The reaction mixture was washed with 10% Na2CO3 solution (2×100 mL) and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with isohexane and increasing the polarity to 30% EtOAc/isohexane to obtain N,N-diethyl-2,3-dimethyl-benzamide as a colourless liquid (1.48 g, 72%).



1H NMR (400 MHz, DMSO-d6): δ 7.20-7.10 (m, 2H), 6.90 (d, J=8 Hz, 1H), 3.70-3.55 (m, 1H), 3.35-3.20 (m, 1H), 3.15-2.90 (m, 2H), 2.25 (s, 3H), 2.07 (s, 3H), 1.17 (t, J=7 Hz, 3H), 0.95 (t, J=7 Hz, 3H).


AnalpH2_MeOH—4 min: Rt 2.75 min; m/z 206 [M+1]+.


The following N,N-diethyl-benzamide derivatives are prepared using analogous procedures.









TABLE 1







N,N-Diethyl-benzamide Derivatives of Formula 2













Mass,





% Yield,


Compound
Reference
Analytical Data
State







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Compound reported by Snieckus et al., 1989
AnalpH2_MeOH_ 4 min: Rt 2.84 min; m/z 226 [M + 1]+
10 g, 77%, colourless oil







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Compound reported by Fujio et al., 2009
AnalpH2_MeOH_ 4 min: Rt 2.63 min; m/z 210 [M + 1]+
1.15 g, 86%, colourless oil







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AnalpH2_MeOH_ 4 min: Rt 2.94 min; m/z 260 [M + 1]+
1.17 g, 92%, colourless oil







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Compound reported by Naoto et al., 2009
AnalpH2_MeOH_ 4 min: Rt 2.80 min; m/z 269 [M + 1]+
4.93 g, 98%, colourless oil









3-Cyclopropyl-N,N-diethyl-2-methyl-benzamide 7



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A solution of 3-bromo-N—N-diethyl-2-methylbenzamide (2.5 g, 9 mmol), cyclopropyl boronic acid (955 mg, 11 mmol), K3PO4 (9.81 g, 46 mmol) and water (10 mL) in toluene (40 mL) was de-gassed using N2 for 1.5 h, Pd(OAc)2 (207 mg, 0.9 mmol) and triphenyl phosphine (42 mg, 0.92 mmol) was added and the reaction mixture degassed for 1 h and heated at 90° C. for 16 h. The reaction mixture was cooled to RT, diluted with EtOAc (40 mL), washed with water (10 mL), dried over Na2SO4 and concentrated in vacuo. The crude compound was purified by silica gel column chromatography eluting with 3% EtOAc/CH2Cl2 to obtain 3-cyclopropyl-N,N-diethyl-2-methyl-benzamide as a pale yellow liquid (1.3 g, 61%).



1H NMR (400 MHz, CDCl3): δ 7.13-7.09 (m, 1H), 7.00-6.98 (m, 2H), 3.91-3.70 (m, 1H), 3.55-3.35 (m, 1H), 3.20-3.05 (m, 2H), 2.34 (s, 3H), 1.95-1.80 (m, 1H), 1.26 (t, J=7 Hz, 3H), 1.02 (d, J=7 Hz, 3H), 0.99-090 (m, 2H), 0.75-0.60 (m, 2H).


AnalpH2_MeOH—4 min: Rt 2.92 min; m/z 232 [M+1]+.


Synthesis of Nitrile Intermediates 3 of Formula 10 (required for Step B, Scheme A)



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Scheme A, Step E (Protocol 1): Synthesis of Amide-Substituted Benzonitriles 10 (Via Acid Coupling)
4-{[(4-Cyano-benzoyl)-methyl-amino]-methyl}-piperidine-1-carboxylic acid tert-butyl ester



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To a stirred solution of 4-cyanobenzoic acid (322 mg, 2.19 mmol) in CH2Cl2 (10 mL) was added TBTU (702 mg, 2.19 mmol) and N,N-diisopropylethylamine (1.14 mL, 6.54 mmol) and the reaction mixture stirred at RT for 10 min. 4-Methylaminomethyl-piperidine-1-carboxylic acid tert-butyl ester (500 mg, 2.19 mmol) in DMF (4 mL) was added and the reaction mixture was stirred at RT for 2 h. The crude material was concentrated in vacuo and purified by silica gel column chromatography, eluting with isohexane and increasing the polarity to 100% EtOAc to afford 4-{[(4-cyano-benzoyl)-methyl-amino]-methyl}-piperidine-1-carboxylic acid tert-butyl ester as a orange solid (700 mg, 89%).


AnalpH2_MeOH—4 min(1): Rt 2.73 min; m/z 358 [M+1]+.


The following nitrile benzamide derivatives are prepared using analogous procedures.









TABLE 2







Amide-substituted Benzonitrile Intermediates 3 of Formula 10











Mass,




%




Yield,


Compound
Analytical Data
State







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AnalpH9_MeOH_ 4 min: Rt 1.87 min; m/z 274 [M + 1]+
1.46 g (92%), yellow semi- solid







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AnalpH9_MeOH_ 4 min: Rt 1.82 min; m/z 258 [M + 1]+
1.27 g (97%), white solid







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AnalpH9_MeOH_ 4 min: Rt 1.89 min; m/z 258 [M + 1]+
1.0 g (99%), white solid







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AnalpH9_MeOH_ 4 min: Rt 2.21 min; m/z 256 [M + 1]+
992 mg (99%), white solid









Scheme B, Step E (Protocol 2): Synthesis of Amide-Substituted Benzonitriles 10 (Via Acid Chloride Coupling)
4-Cyano-N-methyl-N-(1-methyl-piperidin-4-ylmethyl)-benzamide



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4-cyanobenzoylchloride (200 mg, 1.21 mmol) was dissolved in anhydrous CH2Cl2 (4 mL) and cooled to 0° C. Methyl-(1-methyl-piperidin-4-ylmethyl)-amine (172 mg, 1.21 mmol) in anhydrous CH2Cl2 (1 mL) was added followed by N,N-diisopropylethylamine (0.63 mL, 3.62 mmol). The reaction mixture was allowed to warm to RT over 2 h. The reaction mixture was concentrated in vacuo and the crude product purified by reverse phase preparative HPLC-MS to obtain 4-cyano-N-methyl-N-(1-methyl-piperidin-4-ylmethyl)-benzamide as an off-white solid (213 mg, 65%).


AnalpH9_MeOH—4 min(1): Rt 1.77 min; m/z 272 [M+1]+.


Synthesis of Nitrile intermediates 3 of Formula 12 (required for Step B-Scheme A)



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Step F: Synthesis of Amino-Substituted Pyridine-Carbonitrile Derivatives 12
6-(4-Acetyl-piperazin-1-yl)-nicotinonitrile



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6-Chloropyridine-2-carbonitrile (104 mg, 0.75 mmol) and 1-acetylpiperazine (384 mg, 0.75 mmol) in acetonitrile (2.5 mL) were stirred and irradiated using a microwave reactor (300 W, 150° C., 60 min). The reaction mixture was concentrated in vacuo and purified by silica gel column chromatography, eluting with CH2Cl2 and increasing the polarity to 10% MeOH/CH2Cl2 to afford 6-(4-acetyl-piperazin-1-yl)-nicotinonitrile as an off-white solid (172 mg, quant.).


AnalpH2_MeOH—4 min: Rt 1.77 min; m/z 231 [M+1]+.


The following substituted pyridine-carbonitrile derivatives are prepared using analogous procedures.









TABLE 3







Substituted Amino-Pyridine-Carbonitrile Derivatives 3 of formula 12









Compound
Analytical Data
Mass, % Yield, State







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AnalpH9_MeOH_4 min: Rt 2.55 min; m/z 231 [M + 1]+
157 mg, 91%, light brown crystalline solid







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Commercially available
N/A







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Commercially available
N/A







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AnalpH9_MeOH_4 min: Rt 2.30 min; m/z 237 [M + 1]+
94 mg, 17%, yellow solid







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AnalpH2_MeOH_4 min: Rt 3.10 min; m/z not observed
731 mg, 94%, yellow solid







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AnalpH9_MeOH_4 min: Rt 1.80 min; m/z 221 [M + 1]+
496 mg, 88%, yellow solid







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AnalpH9_MeOH_4 min: Rt 1.46 min; m/z 260 [M + 1]+
485 mg, 65%, yellow oil







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AnalpH9_MeOH_4 min: Rt 1.85 min; m/z 233 [M + 1]+
350 mg, 70%, pale yellow solid







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AnalpH9_MeOH_4 min: Rt 2.12 min; m/z 217 [M + 1]+
312 mg, 80%, light brown oil







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AnalpH9_MeOH_4 min: Rt 2.22 min; m/z 231 [M + 1]+
449 mg, 87%, cream solid







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AnalpH9_MeOH_4 min: Rt 2.56 min; m/z 229 [M + 1]+
340 mg, 85%, beige solid







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AnalpH2_MeOH_4 min(3): Rt 0.98 min; m/z 194 [M + 1]+
Used in next step as crude material









Synthesis of Nitrile Intermediates 3 of Formula 14 (required for Step B, Scheme A)



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6-{4-[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-piperazin-1-yl}-nicotinonitrile



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To a solution of 6-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-nicotinonitrile (350 mg, 1.51 mmol) and imidazole (236 mg, 3.47 mmol) in anhydrous DMF (3 mL) was added TBDMS chloride (295 mg, 1.96 mmol) in anhydrous DMF (2 mL) and the reaction mixture stirred for 16 h at RT. The reaction mixture was diluted with EtOAc (5 mL) and washed with water (10 mL) and brine (10 mL). The organic phase was separated, passed through a phase separation cartridge and concentrated in vacuo. The crude residue was purified on silica gel column chromatography eluting with 30% EtOAc/isohexane, and increasing the polarity to 50% EtOAc/isohexane to afford 6-{4-[2-(tert-butyl-dimethyl-silanyloxy)-ethyl]-piperazin-1-yl}-nicotinonitrile as a pale yellow solid (394 mg, 75%).


AnalpH2_MeOH—4 min: Rt 2.17 min; m/z 347 [M+1]+.


The following TBDMS-protected nicotinonitrile derivatives are prepared using analogous procedures.









TABLE 4







Nitrile Intermediates 3 of formula 14













Mass, % Yield,


Compound
Reference
Analytical Data
State







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Intermediate for IQ-219
AnalpH2_MeOH_4 min(3): Rt 3.76 min; m/z 422 [M + 1]+
1.34 g, 88%, white solid









Synthesis of Nitrile Intermediates 3 of Formula 16 (Required for Step B, Scheme A)



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Synthesis of Boc-Protected Amine 9
[2-(tert-Butyl-diphenyl-silanyloxy)-ethyl]-methyl-carbamic acid tert-butyl ester



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(2-Hydroxy-ethyl)-methyl-carbamic acid tert-butyl ester (400 mg, 2.28 mmol), TBDPS chloride (593 μL, 2.28 mmol) and imidazole (342 mg, 5.02 mmol) in DMF (2 mL) were stirred at RT for 12 h. The reaction mixture was diluted with brine and extracted with CH2Cl2. The combined organics were passed through a phase separation cartridge and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with isohexane and increasing the polarity to 50% EtOAc/isohexane to obtain [2-(tert-butyl-diphenyl-silanyloxy)-ethyl]-methyl-carbamic acid tert-butyl ester as a colourless oil (572 mg, 61%).


AnalpH2_MeOH—4 min(1): Rt 3.74 min; m/z 414 [M+1]+.


[2-(tert-Butyl-diphenyl-silanyloxy)-ethyl]-methyl-amine



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[2-(tert-Butyl-diphenyl-silanyloxy)-ethyl]-methyl-carbamic acid tert-butyl ester (572 mg, 1.38 mmol) and 4M HCl/dioxane (3 mL) in CH2Cl2 (5 mL) were stirred at RT for 3 h. The reaction mixture was concentrated in vacuo and the crude material was purified by silica gel column chromatography, eluting with CH2Cl2 and increasing the polarity to 10% MeOH/CH2Cl2 to obtain [2-(tert-butyl-diphenyl-silanyloxy)-ethyl]-methyl-amine as a yellow solid (105 mg, 21%).


AnalpH2_MeOH—4 min(1): Rt 2.41 min; m/z 314 [M+1]+.


Step H: Synthesis of Sulfonamide Derivatives 16
4-{[(4-Cyano-benzenesulfonyl)-methyl-amino]-methyl}-piperidine-1-carboxylic acid tert butyl ester



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To a stirred solution of 4-cyanobenzenesulfonyl chloride (411 mg, 2.2 mmol) in CH2Cl2 (10 mL) was added 4-methylaminomethyl-piperidine-1-carboxylic acid tert-butyl ester (500 mg, 2.2 mmol) and triethylamine (0.91 mL, 6.5 mmol) and the reaction stirred at RT for 2 h after which time silica was added and solvent removed. The crude residue was purified by silica gel chromatography eluting with isohexane, and increasing the polarity to 100% EtOAc to afford 4{[4-cyano-benzenesulfonyl)-methyl-amino]-methyl}-piperidine-1-carboxylic acid tert-butyl ester as a white solid (750 mg, 87%).


AnalpH2_MeOH—4 min(1): Rt 2.91 min; m/z 416 [M+23]+.


The following substituted sulfonamide derivatives are prepared using analogous procedures.









TABLE 5







Sulfonamide Derivatives 3 of formula 16













Mass, % Yield,


Compound
Reference
Analytical Data
State







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AnalpH2_MeOH_ 4 min: Rt 2.72 min; m/z 374 [M + 23]+
669 mg, 96%, white solid







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Commercially available

N/A







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AnalpH2_MeOH_ 4 min(1): Rt 1.24 min; m/z 320 [M + 1]+
300 mg, 94%, cream solid







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AnalpH2_MeOH_ 4 min(1): Rt 2.29 min; m/z not observed
310 mg, quant., white solid







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AnalpH2_MeOH_ 4 min(1): Rt 2.07 min; m/z 268 [M + 1]+
367 mg, quant., cream solid







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AnalpH2_MeOH_ 4 min: Rt 1.80 min; m/z 294 [M + 1]+
560 mg, quant., white solid







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AnalpH2_MeOH_ 4 min: Rt 1.05 min; m/z 308 [M + 1]+
669 mg, quant., white solid







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AnalpH9_MeOH_ 4 min(1): Rt 1.86 min; m/z 294 [M + 1]+
129 mg, 65%, yellow solid







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AnalpH2_MeOH_ 4 min: Rt 0.85 min; m/z 266 [M + 1]+
268 mg, quant., off- white solid







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AnalpH2_MeOH_ 4 min(1): Rt 3.49 min; m/z 479 [M + 1]+
131 mg, 91%, yellow solid









Synthesis of Nitriles 3 of Formula 22



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Scheme F, Step I: Synthesis of Amine Intermediates 18
4-(tert-Butyl-diphenyl-silanyloxy)-piperidine-1-carboxylic acid tert-butyl ester



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To a stirred solution of 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (400 mg, 1.98 mmol) in DMF (2 mL) was added TBDPS chloride (0.52 mL, 1.98 mmol) and imidazole (297 mg, 4.47 mmol) and the reaction stirred at RT for 16 h afterwhich time the reaction mixture was diluted with brine (10 mL), washed with CH2Cl2 (3×25 mL) and the organics combined and dried through a phase separation cartridge and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with EtOAc and increasing the polarity to 30% EtOAc/isohexane to obtain 4-(tert-butyl-diphenyl-silanyloxy)-piperidine-1-carboxylic acid tert-butyl ester as a colourless oil (545 mg, 62%).


AnalpH2_MeOH—4 min(1): Rt 3.92 min; m/z 440 [M+1]+.


The following substituted amine derivatives are prepared using analogous procedures.









TABLE 6







Boc-protected Amine Intermediates 17













Mass, % Yield,


Compound
Code
Analytical Data
State







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IQ-167
AnalpH2_MeOH_ 4 min(3): Rt 3.68 min; m/z 412 [M + 1]+
2.14 g, 90%, colourless oil







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IQ-172
AnalpH2_MeOH_ 4 min(3): Rt 3.75 min; m/z 326 [M − (Boc)]+
1.07 g, 95%, colourless oil









4-(tert-Butyl-diphenyl silanyloxy)-piperidine



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To 4-(tert-butyl-diphenyl-silanyloxy)-piperidine-1-carboxylic acid tert-butyl ester (54 mg, 0.124 mmol) was added 4M HCl/dioxane (2 mL) and CH2Cl2 (5 mL). The reaction mixture was stirred at RT for 2 h. 4M HCl/dioxane (3 mL) added and reaction stirred for a further 1 hr. The reaction mixture was concentrated in vacuo. The crude material was purified by silica gel column chromatography eluting with CH2Cl2 and increasing the polarity to 10% MeOH/CH2Cl2 to obtain 4-(tert-butyl-diphenyl silanyloxy)-piperidine as a cream foam (370 mg, 79%).


AnalpH2_MeOH—4 min(1): Rt 2.54 min; m/z 340 [M+1]+.


The following substituted amine derivatives are prepared using analogous procedures.









TABLE 7







Boc-deprotected Amine Intermediates 18













Mass, % Yield,


Compound
Reference
Analytical Data
State







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Intermediate for IQ-167 or IQ-169
AnalpH2_ MeOH_ 4 min(3): Rt 2.39 min; m/z 312 [M + 1]+
950 mg, 59%, pale oil







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Intermediate for IQ-172
AnalpH2_ MeOH_ 4 min(3): Rt 2.46 min; m/z 326 [M + 1]+
188 mg, 21%, white solid









Scheme F, Step J: Synthesis of Nitrile Intermediates 3 of Formula 22 (Via Bromide Sisplacement)
(3aS,6aR)-5-(4-Cyano-benzyl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester



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To 4-(bromomethyl)benzonitrile (277 mg, 1.41 mmol) was added hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid (300 mg, 1.41 mmol), potassium carbonate (215 mg, 1.55 mmol) and acetone (7 mL) and the reaction mixture stirred for 16 h. The reaction mixture was concentrated in vacuo, dissolved in CH2Cl2 (4 mL) and washed with water (4 mL).


The organic phase was separated and the aqueous layer washed with CH2Cl2 (4 mL). The organic phases were combined, passed through a phase separation cartridge and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with CH2Cl2 and increasing the polarity to 3.5% MeOH/CH2Cl2 to obtain (3aS,6aR)-5-(4-cyano-benzyl)-hexahydro-pyrrolo[3,4-c]pyrrole-2-carboxylic acid tert-butyl ester as a yellow oil (278 mg, 60%).


AnalpH2_MeOH—4 min(1): Rt 1.54 min; m/z 328 [M+1]+.


The following nitrile derivatives are prepared using analogous procedures.









TABLE 8







Nitrile Intermediates 3 of formula 22










Compound
Reference
Analytical Data
Mass, % Yield, State







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AnalpH2_ MeOH_ 4 min(1): Rt 2.53 min; m/z 316 [M + 1]+
630 mg, 100%, white solid







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AnalpH2_ MeOH_ 4 min(1): Rt 1.82 min; m/z 316 [M + 1]+
220 mg, 73%, colourless oil







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AnalpH2_ MeOH_ 4 min(1): Rt 2.51 min; m/z 364 [M + 1]+
197 mg, 85%, colourless oil







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AnalpH2_ MeOH_ 4 min(1): Rt 3.20 min; m/z 330 [M + 1]+
280 mg, 83%, colourless oil







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AnalpH2_ MeOH_ 4 min(1): Rt 1.55 min; m/z 316 [M + 1]+
294 mg, 92%, colourless oil







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AnalpH2_ MeOH_ 4 min: Rt 1.43 min; m/z 314 [M + 1]+
445 mg, 69%, colourless oil







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AnalpH2_ MeOH_ 4 min: Rt 2.54 min; m/z 316 [M + 1]+
550 mg, 85%, colourless oil







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AnalpH2_ MeOH_ 4 min: Rt 1.85 min; m/z 316 [M + 1]+
420 mg, 65%, white solid







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Commercially available

N/A







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Commercially available

N/A







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AnalpH9_ MeOH_ 4 min: Rt 2.51 min; m/z 205 [M + 1]+
760 mg, 80%, pale yellow liquid







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Commercially available

N/A







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AnalpH9_ MeCN_ 4 min; Rt 1.83 min; m/z 230 [M + 1]+
189 mg, 89%, colourless oil







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AnalpH9_ MeCN_ 4 min(1): Rt 1.39 min; m/z 230 [M + 1]+
1.02 g, 85%, orange oil







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AnalpH9_ MeOH_ 4 min(1): Rt 1.81 min; m/z 230 [M + 1]+
150 mg, 64%, colourless oil







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AnalpH9_ MeCN_ 4 min(1): Rt 1.72 min; m/z 256 [M + 1]+
63 mg, 15%, brown oil







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AnalpH2_ MeOH_ 4 min(1): Rt 1.47 min; m/z 230 [M + 1]+
212 mg, 45%, orange glass







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AnalpH9_ MeCN_ 4 min: Rt 1.94 min; m/z 242 [M + 1]+
171 mg, 34%, pale orange oil







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AnalpH9_ MeCN_ 4 min: Rt 1.72 min; m/z 244 [M + 1]+
231 mg, 37%, white solid







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AnalpH2_ MeOH_ 4 min: Rt 0.95 min, m/z 230 [M + 1]+
110 mg, 23%, orange oil







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AnalpH9_ MeCN_ 4 min: Rt 2.19 min; m/z 242 [M + 1]+
68 mg, 13%, pale orange oil







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AnalpH2_ MeOH_ 4 min: Rt 0.34, 0.74 min; m/z 201 [M + 1]+
700 mg, 68%, pale yellow liquid







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AnalpH2_ MeOH_ 4 min: Rt 0.33, 0.57 min; m/z 187 [M + 1]+, 373 [2M + 1]+
700 mg, 73%, pale yellow liquid







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Commercially available

N/A







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AnalpH2_ MeOH_ 4 min(1): Rt 2.69 min; m/z 455 [M + 1]+
360 mg, 80%, yellow glass







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AnalpH2_ MeOH_ 4 min: Rt 2.82 min; m/z 328 [M + 1]+
194 mg, 100%, cream solid







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Intermediate for IQ-167
AnalpH2_ MeOH_ 4 min(3): Rt 2.56 min; m/z 427 [M + 1]+
180 mg, 28%, colourless glass







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Commercially available

N/A







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Commercially available

N/A







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Intermediate for IQ-169
AnalpH2_ MeOH_ 4 min(3): Rt 2.75 min; m/z 445 [M + 1]+
50 mg, 6%, yellow oil







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Intermediate for IQ-173
AnalpH2_ MeOH_ 4 min(3): Rt 0.39 min; m/z 230 [M + 1]+
414 mg, 51%, orange oil







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Intermediate for IQ-174
AnalpH9_ MeOH_ 4 min(2): Rt 0.39 min; m/z 230 [M + 1]+
469 mg, 65%, bright yellow oil







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Commercially available

N/A









Synthesis of Nitriles 3 of Formula 22
Scheme F, Step K: Synthesis of Aryl Bromide Intermediates 21 (Via Amine dialkylation)
1-[1-(4-Bromo-phenyl)-1-methyl-ethyl]-4-(toluene-4-sulfonyl)-piperazine



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To a solution of 1-(4-bromo-phenyl)-1-methyl-ethylamine (400 mg, 1.84 mmol) in diisopropylethylamine (4 mL) was added N,N-bis(2-chloroethyl)-4-methylbenzene sulphonamide (500 mg, 1.68 mmol) and the reaction subjected to microwave irradiation at 150° C. for 9 h afterwhich time the reaction was concentrated in vacuo and the crude residue purified by reverse phase preparative HPLC-MS to afford 1-[1-(4-bromo-phenyl)-1-methyl-ethyl]-4-(toluene-4-sulfonyl)-piperazine as a peach solid (375 mg, 47%).


AnalpH2_MeOH—4 min(1): RT 3.04 min; m/z 437/439 [M+1]+.


Scheme F, Step L: Synthesis of Nitrile Intermediates 22
4-{1-Methyl-1-[4-(toluene-4-sulfonyl)-piperazin-1-yl]-ethyl}-benzonitrile



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To a solution of 1-[1-(4-bromo-phenyl)-1-methyl-ethyl]-4-(toluene-4-sulfonyl)-piperazine (200 mg, 0.45 mmol) in DMF (3 mL) was added zinc cyanide (64.41 mg, 0.54 mmol) and tetrakis(triphenylphosphine)palladium(0) (52 mg, 0.045 mmol) and the reaction mixture degassed for 10 min under N2. The reaction mixture was then subjected to microwave irradiation for 30 min at 180° C., afterwhich time the reaction was diluted with 1:1 CH2Cl2/EtOAc (20 mL), washed with water (2×10 mL), passed through a phase separation cartridge and concentrated in vacuo. The crude residue was purified by reverse phase preparative HPLC-MS to afford 4-{1-methyl-1-[4-(toluene-4-sulfonyl)-piperazin-1-yl]-ethyl]-benzonitrile as a cream solid (70 mg, 47%).


AnalpH2_MeOH—4 min(1): Rt 2.78 min; m/z 384 [M+1]+.


Scheme F, Step M: Synthesis of Nitrile Intermediates 22 (Via BOC Protection)
(4-Cyano-benzyl)-methyl-carbamic acid tert-butyl ester



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To 4-[(methylamine)methyl]benzonitrile (1 g, 6.8 mmol) in CH2Cl2 (50 mL) was added DMAP (0.93 g, 7.6 mmol), di-tert-butyl dicarbonate (1.7 g, 7.6 mmol) and the reaction stirred for 48 h at RT. The reaction mixture was washed with saturated, aqueous NaHCO3 and brine. The organic phase was separated and concentrated in vacuo. The crude residue was purified on silica gel chromatography eluting with isohexane, and increasing the polarity to 20% EtOAc/isohexane to afford (4-cyano-benzyl)-methyl-carbamic acid tert-butyl ester as a colourless oil (1.48 g, 89%).


AnalpH2_MeOH—4 min: Rt 2.75 min; m/z 247 [M+1]+.


Scheme F, Step AO: Synthesis of Nitrile Intermediates 22 (Via Reductive Amination)
4-[3-(tert-Butyl-diphenyl-silanyloxy)-3-methyl-azetidin-1-ylmethyl]-benzonitrile



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To a stirred solution of 4-formylbenzonitrile (68 mg, 0.51 mmol) and 3-(tert-Butyl-diphenyl-silanyloxy)-3-methyl-azetidine hydrochloride (188 mg, 0.51 mmol) in 1:1 MeOH/DMF (26 mL) was added acetic acid (catalytic). The reaction mixture was stirred under N2 at 0° C. for 1 h. Sodium cyanoborohydride (1M in THF, 0.6 mL, 0.57 mmol) was added and the reaction mixture stirred at RT, under N2 for 18 h. The reacton mixture was concentrated in vacuo, the residue suspended in H2O (10 mL), washed with CH2Cl2 (2×10 mL) and the solution passed through a phase separation cartridge. The combined organic layers were concentrated in vacuo and the crude residue purified by silica gel chromatography eluting with 100% isohexane and increasing the polarity 100% EtOAc to afford 4-[3-(tert-butyl-diphenyl-silanyloxy)-3-methyl-azetidin-1-ylmethyl]-benzonitrile as a colourless oil (196 mg, 86%).


AnalpH2_MeOH—4 min(3): Rt 2.71 min; m/z 441 [M+1]+.


Synthesis of Nitrile Intermediates 3 of Formula 26 (Required for Step B, Scheme A)



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Scheme G, Step N: Mesylation of Alcohol 24
4-Cyano phenethyl methanesulfonate



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To a solution of 4-(2-hydroxy-ethyl)-benzonitrile (2 g, 13.6 mmol) in CH2Cl2 (10 mL) was added Et3N (6.8 mL, 47.52 mmol) and mesyl chloride (1.4 mL, 17.63 mmol) at 0° C. and stirred for 2 h. The reaction mixture was diluted with CH2Cl2 (30 mL), washed with saturated NaHCO3 solution (2×10 mL), the organic layer was dried over Na2SO4, filtered and concentrated in vacuo to obtain 4-cyano phenethyl methanesulfonate (3 g) as a pale yellow gummy liquid. The crude compound was used for the next step without further purification.


Rf: 0.6 (50% EtOAc/petroleum ether 60-80).


Scheme G, Step O: Synthesis of Amines (Via Mesylate Displacement)
4-(2-Morpholinoethyl)benzonitrile



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To a stirred solution of 4-cyano phenethyl methanesulfonate (6.04 mmol) in CH2Cl2 (10 mL) at 0° C. was added morpholine (3.5 g, 40.22 mmol) and heated 50° C. for 16 h. The reaction mixture was diluted with CH2Cl2 (100 mL), washed with saturated NaHCO3 solution (2×10 mL), the organic layer was dried over Na2SO4 and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with 3% MeOH/CHCl3 to obtain 4-(2-morpholinoethyl)benzonitrile as a pale yellow solid (700 mg, 48%).



1H NMR (400 MHz, CDCl3): δ 7.58 (d, J=8 Hz, 2H), 7.30 (d, J=8 Hz, 2H), 3.72 (t, J=4.8 Hz, 4H), 2.85 (t, J=8 Hz, 2H), 2.61-2.49 (6H, m).


AnalpH9_MeOH—4 min: Rt 2.20 min; m/z 217 [M+1]+.


The following nitrile derivatives are prepared using analogous procedures.









TABLE 9







Nitrile Intermediates 3 of formula 26









Compound
Analytical Data
Mass, % Yield, State







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AnalpH9_ MeOH_4 min: Rt 2.39 min; m/z 201 [M + 1]+
Pale yellow solid







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AnalpH9_ MeOH_4 min: Rt 2.62 min; m/z 256 [M + 1]+
Pale yellow solid









Scheme a, Step B (Protocol 1): Synthesis of Boc-Protected 2H-Isoquinolin-1-One Derivatives of Formula 4
4-({Methyl-[4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoyl]-amino}-methyl)-piperidine-1-carboxylic acid tert-butyl ester (IQ-092)



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N,N-Diethyl-2,3-dimethyl-benzamide (200 mg, 0.97 mmol) was dissolved in anhydrous THF (4 mL) under a N2 and cooled to −78° C. n-BuLi (2.5M in n-hexanes, 0.82 mL, 2.04 mmol) was added dropwise to yield a deep red coloured solution and the reaction mixture was stirred at −78° C. for 30 minutes. 4-{[(4-Cyano-benzoyl)-methyl-amino]-methyl}-piperidine-1-carboxylic acid tert-butyl ester (348 mg, 0.97 mmol) was dissolved in anhydrous THF (4 mL) and added dropwise, and the reaction stirred at −78° C. for 2 h. The reaction mixture was quenched with ice/water, allowed to warm to RT and extracted with CH2Cl2 and EtOAc. The combined organic phase was passed through a phase separation cartridge and concentrated in vacuo. The crude compound was triturated with isohexane/diethyl ether (80:20), the solid filtered and dried in vacuo to give 4-({methyl-[4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoyl]-amino}-methyl)-piperidine-1-carboxylic acid tert-butyl ester as a light beige solid (171 mg, 36%).


AnalpH2_MeOH_QC(Sunfire1): Rt 7.81 min; m/z 490 [M+1]+.


4-(5-Chloro-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(1-methyl-piperidin-4-ylmethyl)-benzamide (IQ-091)



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3-Chloro-N,N-diethyl-2-methyl-benzamide (150 mg, 0.66 mmol) was dissolved in anhydrous THF (2 mL) under N2 and cooled to −78° C. n-BuLi (2.5M in n-hexanes, 558 μL, 1.39 mmol) was added dropwise and the reaction mixture was stirred at −78° C. for 30 minutes. 4-Cyano-N-(1-methyl-piperidin-4-ylmethyl)-benzamide (180 mg, 0.66 mmol) in anhydrous THF (2 mL) was added dropwise to the reaction mixture and stirred at −78° C. continued for 1 h. The reaction mixture was poured into ice/water, allowed to warm to RT and extracted with CH2Cl2 (×3) and the organic phase dried (MgSO4). The solution was filtered and concentrated in vacuo. The crude material was purified by reverse phase preparative HPLC-MS to afford 4-(5-chloro-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(1-methyl-piperidin-4-ylmethyl)-benzamide as a white solid (37 mg, 13%).



1H NMR (400 MHz, DMSO-d6): δ11.91 (br s, 1H), 8.25 (s, formic acid, 1H), 8.21 (d, J=8. Hz, 1H), 7.9 (dd, J=8 Hz, 1H), 7.85 (d, J=8 Hz, 2H), 7.53 (d, J=8 Hz, 1H), 7.51 (t, J=8 Hz, 1H), 7.46 (d, J=8 Hz, 1H), 6.97 (s, 1H), 3.42-3.38 (m, 1H), 3.20-3.12 (m, 1H), 2.95 (s, 1H), 2.90 (s, 2H), 2.82-2.78 (m, 1H), 2.68-2.64 (m, 1H), 2.18 (s, 2H), 2.09 (s, 1H), 1.86-1.92 (m, 1H), 1.79-1.81 (m, 1H), 1.68-1.65 (m, 2H), 1.49-1.42 (m, 1H), 1.30-1.23 (m, 1H), 0.90-0.79 (m, 1H).


AnalpH2_MeOH_QC: Rt 5.70 min; m/z 424 [M+1]+.









TABLE 10







2H-Isoquinolin-1-one Derivatives of Formula 4













Mass, % Yield,


Compound
Code
Analytical Data
State







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IQ-145
AnalpH2_ MeOH_QC (Sunfire 1): Rt 8.08 min; m/z 510 [M + 1]+
142 mg, 31%, light beige solid







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IQ-101
AnalpH2_ MeOH_QC: Rt 5.21 min; m/z 382 [M + 1]+
10 mg, 3%, pale yellow solid







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IQ-102
AnalpH2_ MeOH_ QC: Rt 5.08 min; m/z 407 [M + 1]+
24 mg, 5%, beige solid







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IQ-103
AnalpH2_ MeOH_QC: Rt 5.33 min; m/z 410 [M + 1]+
5 mg, 3%, white solid







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IQ-104
AnalpH2_ MeOH_QC: Rt 4.93 min; m/z 362 [M + 1]+
27 mg, 9%, white solid 1H NMR (400 MHz, DMSO-d6): δ 11.61 (br s, 1H), 8.09 (d, J = 7.8 Hz, 1H), 7.90 (d, J = 8.6 Hz, 2H), 7.58 (d, J = 7.3 Hz, 1H), 7.50 (d, J = 8.6 Hz, 2H), 7.4 (t, J = 7.8 Hz, 1H), 6.92 (s, 1H), 3.63 (br s, 2H), 3.38 (br s, 2H), 2.57 (s, 3H), 2.34 (br s, 4H), 2.21 (s, 3H).







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IQ-001
AnalpH2_ MeOH_QC: Rt 8.49 min; m/z 279 [M + 1]+
143 mg, 64%, pale yellow solid







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IQ-013
AnalpH2_ MeOH_QC: Rt 8.84 min; m/z 420 [M + 1]+
123 mg, 43%, yellow solid







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IQ-010
AnalpH2_ MeOH_ QC: Rt 9.14 min; m/z 440 [M + 1]+
231 mg, 60%, cream solid







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IQ-042
AnalpH2_ MeOH_4 min: Rt 5.21 min; m/z 349 [M + 1]+
46 mg, 26%, off- white solid







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IQ-041
AnalpH2_ MeOH_4 min: Rt 5.30 min; m/z 333 [M + 1]+
30 mg, 9%, off- white solid







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IQ-137
AnalpH2_ MeOH_4 min: Rt 5.55 min; m/z 388 [M + 1]+
12 mg, 6%, white solid







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IQ-131
AnalpH2_ MeOH_QC: Rt 5.74 min; m/z 377 [M + 1]+
22 mg, 10%, pale brown gum







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IQ-132
AnalpH2_ MeOH_QC: Rt 5.69 min; m/z 387 [M + 1]+
1.23 g, 86%, cream solid







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IQ-128
AnalpH2_ MeOH_QC: Rt 5.59 min; m/z 343 [M + 1]+
86 mg, 32%, beige solid







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IQ-129
AnalpH2_ MeOH_QC: Rt 5.21 min; m/z 327 [M + 1]+
135 mg, 52%, beige solid







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IQ-130
AnalpH2_ MeOH_QC: Rt 5.27 min; m/z 323 [M + 1]+
101 mg, 40%, pale yellow solid







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IQ-133
AnalpH2_ MeOH_QC: Rt 5.37 min; m/z 365 [M + 1]+
230 mg, 63%, white solid







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IQ-134
AnalpH2_ MeOH_QC: Rt 5.53 min; m/z 349 [M + 1]+
42 mg, 24%, off- white solid







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IQ-135
AnalpH2_ MeOH_QC: Rt 8.84 min; m/z 369 [M + 1]+
40 mg, 22%, off- white solid







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IQ-003
AnalpH2_ MeOH_QC: Rt 5.16 min; m/z 335 [M + 1]+
14 mg, 4%, off- white solid







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IQ-002-1
AnalpH2_ MeOH_QC: Rt 5.45 min; m/z 355 [M + 1]+
196 mg, 84%, pale yellow solid







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IQ-153
AnalpH9_ MeOH_QC: Rt 8.51 min; m/z 421 [M + 1]+
24 mg, 6%, yellow solid







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IQ-136
AnalpH2_ MeOH_QC: Rt 8.79 min; m/z 441 [M + 1]+
126 mg, 29%, off- white solid







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IQ-144
AnalpH2_ MeOH_QC: Rt 5.55 min; m/z 369 [M + 1]+
5.5 mg, 4%, white solid







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IQ-139
AnalpH2_ MeOH_QC: Rt 9.37 min; m/z 459 [M + 1]+
139 mg, 34%, orange solid







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IQ-020
AnalpH2_ MeOH_QC: Rt 9.03 min; m/z 439 [M + 1]+
63 mg, 15%, cream solid







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IQ-021
AnalpH2_ MeOH_QC: Rt 5.37 min; m/z 353 [M + 1]+
8 mg, 3%, cream solid







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IQ-022
AnalpH2_ MeOH_QC: Rt 5.69 min; m/z 373 [M + 1]+
56 mg, 23%, cream solid







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IQ-019
AnalpH2_ MeOH_QC: Rt 5.59 min; m/z 412 [M + 1]+
49 mg, 26%, off- white solid







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IQ-018
AnalpH2_ MeOH_QC: Rt 5.10 min; m/z 392 [M + 1]+
50 mg, 28%, pale yellow solid







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AnalpH2_ MeOH_ 4 min: Rt 2.68 min; m/z 499 [M + 1]+
222 mg, 99%, yellow solid







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AnalpH2_ MeOH_4 min: Rt 2.58 min; m/z 479 [M + 1]+
214 mg, 99%, orange solid







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IQ-009
AnalpH2_ MeOH_QC: Rt 5.29 min; m/z 369 [M + 1]+
25.5 mg, 15%, tan solid







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IQ-008
AnalpH2_ MeOH_QC: Rt 4.77 min; m/z 349 [M + 1]+
28 mg, 18%, pale yellow solid







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IQ-007
AnalpH2_ MeOH_QC: Rt 5.51 min; m/z 384 [M + 1]+
72 mg, 42%, yellow solid 1H NMR (400 MHz, DMSO-d6): δ 11.72 (br s, 1H), 8.54 (d, J = 2.8 Hz, 1H), 8.16 (dt, J = 7.6 Hz, 1H), 7.92 (dd, J = 9.1 Hz, 1H), 7.84 (dd, J = 7.6 Hz, 1H), 7.42 (t, J = 7.8 Hz, 1H), 6.94 (d, J = 8.8 Hz, 1H), 6.82 (s, 1H), 4.4 (br d, J = 13.1 Hz, 2H), 2.91 (t, J = 11.9 Hz, 2H), 2.39-2.38 (m, 1H), 2.19 (s, 6H), 1.83 (br d, J = 12.6 Hz, 2H), 1.39-1.29 (m, 2H).







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IQ-006
AnalpH2_ MeOH_QC: Rt 5.09 min; m/z 364 [M + 1]+
26 mg, 16%, pale yellow solid







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IQ-005
AnalpH2_ MeOH_QC: Rt 7.39 min; m/z 363 [M + 1]+
24.5 mg, 8%, pale yellow solid







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IQ-004
AnalpH2_ MeOH_QC: Rt 5.45 min; m/z 362 [M + 1]+
53 mg, 33%, pale yellow solid







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IQ-114
AnalpH2_ MeOH_QC (Sunfire 1): Rt 8.21 min; m/z 546 [M + 1]+
210 mg, 43%, white solid







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IQ-113
AnalpH2_ MeOH_QC (Sunfire): Rt 7.98 min; m/z 526 [M + 1]+
230 mg, 45%, cream solid







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IQ-141
AnalpH2_ MeOH_QC: Rt 8.71 min; m/z 504 [M + 1]+
189 mg, 57%, yellow solid







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IQ-140
AnalpH2_ MeOH_QC: Rt 8.42 min; m/z 484 [M + 1]+
107 mg, 31%, yellow solid







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IQ-108
AnalpH2_ MeOH_QC: Rt 7.56 min; m/z 343 [M + 1]+
32.5 mg, 12%, cream solid







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IQ-119
AnalpH2_ MeOH_QC(1): Rt 5.71 min; m/z 452 [M + 1]+
83 mg, 20%, cream solid







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IQ-118
AnalpH2_ MeOH_QC(1): Rt 7.98 min; m/z 427 [M + 1]+
120 mg, 28%, pale yellow solid







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IQ-117
AnalpH2_ MeOH_QC(1): Rt 5.38 min; m/z 400 [M + 1]+
193 mg, 43%, pale yellow solid







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IQ-126
AnalpH2_ MeOH_QC: Rt 5.85 min; m/z 446 [M + 1]+
163 mg, 56%, yellow yellow solid







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IQ-125
AnalpH2_ MeOH_QC: Rt 5.55 min; m/z 426 [M + 1]+
104 mg, 33%, yellow yellow solid







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IQ-110
AnalpH2_ MeOH_QC: Rt 5.87 min; m/z 446 [M + 1]+
130 mg, 48%, white solid







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IQ-111
AnalpH2_ MeOH_QC: Rt 5.72 min; m/z 440 [M + 1]+
47 mg, 15%, white solid







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IQ-112
AnalpH2_ MeOH_QC: Rt 6.02 min; m/z 461 [M + 1]+
52 mg, 18%, white solid 1H NMR (400 MHz, DMSO-d6): δ 11.99 (br s, 1H), 8.23-8.21 (m, 1H), 8.21 (s, formic acid CHO, 0.4H), 8.06-8.03 (m, 2H), 7.92-7.88 (m, 3H), 7.54 (t, J = 7.6 Hz, 1H), 7.02 (s, 1H), 2.84 (d, J = 7.6 Hz, 2H), 2.80- 2.74 (m, 2H), 2.70 (s, 3H), 2.18 (s, 3H), 1.92-1.86 (m, 2H), 1.65-1.50 (m, 2H), 1.59-1.50 (m, 1H), 1.21-1.10 (m, 2H).







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IQ-109
AnalpH2_ MeOH_QC: Rt 5.58 min; m/z 426 [M + 1]+
10 mg, 4%, white solid







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IQ-122
AnalpH2_ MeOH_QC: Rt 5.80 min; m/z 418 [M + 1]+
142 mg, 68%, cream solid







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IQ-121
AnalpH2_ MeOH_QC: Rt 5.49 min; m/z 398 [M + 1]+
77 mg, 39%, cream solid







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AnalpH2_ MeOH_QC(1): Rt 9.80 min; m/z 611 [M + 1]+
3.8 mg, 2%, white solid







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IQ-031
AnalpH2_ MeOH_QC: Rt 5.44 min; m/z 313 [M + 1]+
138 mg, 67%, cream solid







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IQ-030
AnalpH2_ MeOH_QC: Rt 4.83 min; m/z 347 [M + 1]+
2 mg, 1% yellow solid







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IQ-032
AnalpH2_ MeOH_QC: Rt 4.90 min; m/z 297 [M + 1]+
175.1 mg, 91% cream solid







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IQ-034
AnalpH2_ MeOH_QC: Rt 4.94 min; m/z 293 [M + 1]+
133 mg, 58% pale yellow solid







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IQ-147
AnalpH2_ MeOH_QC(1): Rt 8.32 min; m/z 448 [M + 1]+
36 mg, 8%, white solid







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IQ-089
AnalpH2_ MeOH_QC(1): Rt 6.02 min; m/z 460 [M + 1]+
11.8 mg, 31%, off- white foam







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IQ-069
AnalpH2_ MeOH_QC(1): Rt 6.51 min; m/z 438 [M + 1]+
111 mg, 51%, pink solid







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IQ-066
AnalpH2_ MeOH_QC(1): Rt 6.45 min; m/z 448 [M + 1]+
131 mg, 43%, cream solid







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IQ-064
AnalpH2_ MeOH_QC(1): Rt 7.05 min; m/z 496 [M + 1]+
28 mg, 7.3%, white solid







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IQ-061
AnalpH2_ MeOH_QC(1): Rt 8.89 min; m/z 462 [M + 1]+
180 mg, 46%, cream solid







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IQ-060
AnalpH2_ MeOH_QC(1): Rt 6.03 min; m/z 448 [M + 1]+
160 mg, 38%, off- white solid







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IQ-085
AnalpH2_ MeOH_QC: Rt 6.01 min; m/z 446 [M + 1]+
37 mg, 18%, white solid







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IQ-143
AnalpH2_ MeOH_QC: Rt 7.21 min; m/z 448 [M + 1]+
98 mg, 30%, white solid







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IQ-142
AnalpH2_ MeOH_QC: Rt 6.42 min; m/z 448 [M + 1]+
164 mg, 50%, white solid







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IQ-047
AnalpH2_ MeOH_QC: Rt 6.82 min; m/z 460 [M + 1]+
38 mg, 16%, white solid







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IQ-044
AnalpH2_ MeOH_QC: Rt 6.91 min; m/z 454 [M + 1]+
138 mg, 46%, white solid







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IQ-040
AnalpH2_ MeOH_QC: Rt 6.41 min; m/z 434 [M + 1]+
395 mg, 46%, white solid







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IQ-058
AnalpH2_ MeOH_QC(1): Rt 7.21 min; m/z 452 [M + 1]+
120 mg, 39%, pale yellow solid







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IQ-039
AnalpH2_ MeOH_QC: Rt 5.23 min; m/z 337 [M + 1]+
29 mg, 11%, pale yellow solid







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IQ-038
AnalpH2_ MeOH_QC: Rt 5.47 min; m/z 348 [M + 1]+
615 mg, 59%, white solid 1H NMR (400 MHz, DMSO-d6): δ 11.56 (br s, 1H), 8.07 (d, J = 8.1 Hz, 1H, J = 7.78 (d, J = 8.3 Hz, 2H), 7.56 (d, J = 6.8 Hz, 1H), 7.41 (d, J = 8.3 Hz, 2H), 7.37 (t, J = 7.6 Hz, 1H), 6.85 (s, 1H), 3.51 (s, 2H), 2.56 (s, 3H), 2.33 (br s, 8H), 2.16 (s, 3H)







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IQ-048
AnalpH2_ MeOH_QC: Rt 5.77 min; m/z 362 [M + 1]+
44 mg, 15%, cream solid







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IQ-056
AnalpH9_ MeOH_QC (Sunfire 1): Rt 7.22 min; m/z 382 [M + 1]+
16 mg, 4%, light beige solid







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IQ-065
AnalpH2_ MeOH_QC(1): Rt 4.49 min; m/z 362 [M + 1]+
14 mg, 5%, cream solid







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IQ-088
AnalpH2_ MeOH_QC (Sunfire): Rt 2.93 min; m/z 388 [M + 1]+
12 mg, 13%, white solid







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IQ-043
AnalpH2_ MeOH_QC: Rt 5.75 min; m/z 368 [M + 1]+
92 mg, 38%, white solid







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IQ-054
AnalpH2_ MeOH_QC (Sunfire): Rt 6.13 min; m/z 362 [M + 1]+
19 mg, 6%, pale orange solid







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IQ-087
AnalpH2_ MeOH_QC: Rt 4.69 min; m/z 374 [M + 1]+
17 mg, 16%, orange solid







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IQ-053
AnalpH2_ MeOH_QC: Rt 4.09 min; m/z 376 [M + 1]+
14 mg, 3%, white solid 1H NMR (400 MHz, DMSO-d6): δ 11.53 (br s, 1H), 8.30 (s, formic acid CHO, 0.5H), 8.07 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 6.8 Hz, 1H), 7.41 (d, J = 8.4 Hz, 2H), 7.37 (d, J = 8.0 Hz, 1H), 6.85 (s, 1H), 3.50 (s, 2H), 2.86-2.83 (m, 2H), 2.56 (s, 3H), 2.18 (s, 6H), 2.12-2.04 (m, 1H), 1.98-1.92 (m, 2H), 1.74-1.71 (m, 2H), 1.44-1.34 (m, 2H).







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IQ-052
AnalpH2_ MeOH_QC: Rt 570 min; m/z 362 [M + 1]+
37 mg, 21%, white solid







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IQ-050
AnalpH2_ MeOH_QC: Rt 5.60 min; m/z 374 [M + 1]+
9 mg, 8.5%, beige solid







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IQ-046
AnalpH2_ MeOH_QC: Rt 5.81 min; m/z 374 [M + 1]+
154 mg, 32%, yellow yellow solid







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IQ-037
AnalpH2_ MeOH_QC: Rt 5.27 min; m/z 665 [2M + 1]+
79 mg, 30%, beige solid







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IQ-036
AnalpH2_ MeOH_QC: Rt 5.12 min; m/z 319 [M + 1]+
172 mg, 68%, yellow/orange solid







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IQ-035
AnalpH2_ MeOH_QC: Rt 5.11 min; m/z 335 [M + 1]+
51 mg, 19%, white solid







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AnalpH2_ MeOH_ 4 min(1): Rt 2.86 min m/z 587 [M + 1]+.
213 mg, 46%, white solid







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IQ-148
AnalpH2_ MeOH_QC(1): Rt 7.99 min m/z 516 [M + 1]+.
34 mg (29%) White solid







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IQ-155
AnalpH2_ MeOH_QC: Rt 7.77 min m/z 460 [M + 1]+.
32 mg, 23%, white solid







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Intermediate for IQ-167
AnalpH2_ MeOH_ 4 min(3): Rt 2.77 min; m/z 559 [M + 1]+
210 mg, 89%, off- white solid







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IQ-168
AnalpH2_ MeOH_QC(2): Rt 4.74 min; m/z 330 [M + 1]+
8.2 mg, 52 %, pale yellow solid 1H NMR (400 MHz, DMSO-d6): δ 11.59 (br s, 1H), 8.08 (d, J = 8.0 Hz, 1H), 7.85 (d, J = 8.0 Hz, 2H), 7.57 (d, J = 6.8 Hz, 1H), 7.48 (s, 1H), 7.40-7.36 (m, 3H), 7.24 (s, 1H), 6.87 (s, 1H), 5.35 (s, 2H), 2.56 (s, 3H), 2.45 (s, 3H).







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Intermediate for IQ-169
AnalpH2_ MeOH_4 min(3): Rt 2.89 min; m/z 577 [M + 1]+
35 mg, 54%, cream solid







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IQ-174
AnalpH2_ MeOH_QC(1): Rt 4.77 min; m/z 348 [M + 1]+
119.5 mg, 43%, off-white solid







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IQ-182
AnalpH2_ MeOH_QC(2): Rt 7.52 min; m/z 316 [M + 1]+
61 mg, 10% off-white solid 1H NMR (400 MHz, DMSO-d6): δ 11.57 (br s, 1H), 8.07 (d, J = 8.0 Hz, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.79 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 7.2 Hz, 1H), 7.49 (d, J = 1.6 Hz, 1H), 7.37 (t, J = 7.6 Hz, 1H), 7.32 (d, J = 8.0 Hz, 2H), 6.84 (s, 1H), 6.30 (t, J = 2.0 Hz, 1H), 5.41 (s, 2H), 2.55 (s, 3H).









Scheme A, Step B (Protocol 2): Synthesis of Boc-Protected 2H-Isoquinolin-1-One Derivatives of Formula 4 Via Reverse Addition Protocol
N-Methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(1-methyl-piperidin-4-yl)-benzamide (IQ-100)



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To a solution of N,N-diethyl-2,3-dimethyl-benzamide (578 mg, 2.82 mmol) in anhydrous THF (3 mL) under N2 at −78° C. was added dropwise n-BuLi (2.5M in n-hexanes, 2.4 mL, 5.92 mmol) to give a deep red solution. The reaction mixture was stirred at −78° C. for 30 minutes. The reaction mixture was transferred dropwise, via syringe, to a reaction vessel containing 4-cyano-N-methyl-N-(1-methyl-piperidin-4-yl)-benzamide (725 mg, 2.82 mmol) in anhydrous THF (5 mL) at −78° C. and under N2. The reaction mixture was stirred at −78° C. for 3.5 h. Water (10 mL) was added and the reaction mixture was extracted with EtOAc (10 mL) and CH2Cl2 (10 mL). The combined organic layers concentrated in vacuo and the resultant solid was triturated with 2:1 isohexane/EtOAc, filtered and dried in vacuo. The crude material was purified by silica gel column chromatography, eluting with CH2Cl2 and increasing the polarity to 15% MeOH/CH2Cl2 to afford N-methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(1-methyl-piperidin-4-yl)-benzamide as a white solid (487 mg, 44%).



1H NMR (400 MHz, DMSO-d6): δ11.80-11.41 (brs, 1H), 8.10 (d, J=8 Hz, 1H), 7.89 (d, J=8 Hz, 2H), 7.58 (d, J=7 Hz, 1H), 7.48 (d, J=8 Hz, 2H), 7.39 (t, J=7 Hz, 1H), 6.93 (s, 1H), 3.31 (s, 3H), 2.96-2.70 (m, 5H), 2.58 (s, 3H), 2.23-1.96 (m, 3H), 1.93-1.71 (brs, 2H), 1.71-1.53 (br s, 2H).


AnalpH2_MeOH_QC(Sunfire): Rt 4.29 min; m/z 390 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 11







2H-isoquinolin-1-one derivatives of Formula 4













Mass, % Yield,


Compound
Code
Analytical Data
State







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IQ-105
AnalpH2_MeOH_QC(1): Rt 4.97 min; m/z 388 [M + 1]+
2.7 mg, 3%, white solid







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IQ-106
AnalpH2_MeOH_QC(1): Rt 4.98 min; m/z 390 [M + 1]+
4 mg, 2%, white solid







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IQ-171
AnalpH2_MeOH_QC(1): Rt 7.41 min; m/z 317 [M + 1]+
8.5 mg, 4%, white solid







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Intermediate for IQ-219
AnalpH2_MeOH_4 min(3): Rt 2.59 min; m/z 554 [M + 1]+
Used in next step as crude material







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Intermediate for IQ-172
AnalpH2_MeOH_4 min(3): Rt 2.86 min; m/z 573 [M + 1]+
63 mg, 27%, white solid







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IQ-173
AnalpH2_MeOH_QC(1): Rt 4.46 min; m/z 363 [M + 1]+
41 mg, 23%, white solid









Scheme A, Step B (Protocol 3): Synthesis of Boc-Protected 2H-Isoquinolin-1-One Derivatives of Formula 4 (LDA Protocol)
4-[4-(5-Bromo-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzyl]-piperazine-1-carboxylic acid tert-butyl ester (IQ-149)



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To a stirred solution of N,N-diisopropylamine (1.56 mL, 11.10 mmol) in THF (5 mL) under N2 at −78° C. was added n-BuLi (2.5M in hexanes) (4.44 mL, 11.10 mmol) and the reaction stirred at −78° C. for 20 min, after which time a solution of 3-bromo-N,N-diethyl-2-methyl-benzamide (1 g, 3.70 mmol) in THF (5 mL) was added, and the reaction stirred at −78° C. for 30 minutes. A solution of 4-(4-cyano-benzyl)-piperazine-1-carboxylic acid tert-butyl ester (1.15 g, 3.70 mmol) in THF (5 mL) was added and the reaction stirred at −78° C. for 2 h. The reaction was quenched with ice and water, EtOAc added and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with isohexane and increasing the polarity to 100% EtOAc to afford 4-[4-(5-bromo-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzyl]-piperazine-1-carboxylic acid tert-butyl ester as a cream solid (1.22 g, 66%).


AnalpH2_MeOH_QC: Rt 6.94 min; m/z 498 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 12







2H-isoquinolin-1-one derivatives of Formula 4










Compound
Code
Analytical Data
Mass, % Yield, State







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IQ-033
AnalpH2_MeOH_QC: Rt 5.40 min; m/z 357 [M + 1]+
56 mg, 15% cream solid







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IQ-156
AnalpH2_MeOH_4 min: Rt 1.85 min; m/z 454 [M + 1]+.
122 mg, 71%, pale yellow solid









4-[4-(5-Ethyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzyl]-piperazine-1-carboxylic acid tert-butyl ester (IQ-151)



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To a stirred solution of 4-[4-(5-Bromo-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzyl]-piperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.4 mmol) in anhydrous THF (4 mL) under N2 was added dichlorobis(tri-o-tolylphosphine)palladium(II) (14 mg, 0.02 mmol), CeCl3 (99 mg, 0.4 mmol) and AlEt3 (1M in hexanes, 1.5 mL, 1.2 mmol) and the reaction stirred at RT for 4 h. The reaction was quenched with ice, diluted with 0.5M aqueous Rochelle's salts (30 mL) and extracted with EtOAc (3×40 mL). The combined organics were washed with Rochelle's salts (2×50 mL), brine (50 mL), dried over MgSO4, filtered and concentrated in vacuo and the crude residue purified by reverse phase preparative HPLC-MS to afford 4-[4-(5-ethyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzyl]-piperazine-1-carboxylic acid tert-butyl ester as an orange solid (83 mg, 61%).


AnalpH2_MeOH_QC(1):Rt 5.13 min; m/z 446 [M+1]+.


5-Methyl-3-[4-(2-methylamino-ethoxy)-phenyl]-2H-isoquinolin-1-one (IQ-127)



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1-Chloro-ethyl chloroformate (97 mg, 0.68 mmol) in 1,2-dichloroethane (0.3 mL) was added to a solution of 3-[4-(2-dimethylamino-ethoxy)-phenyl]-5-methyl-2H-isoquinolin-1-one (35 mg, 0.109 mmol) in 1,2-dichloroethane (0.6 mL) at cooled to 0° C., and stirred for 10 min. The reaction mixture was irradiated using a microwave (300 W, 180° C., 15 min) then concentrated in vacuo and EtOH (0.8 mL) added. The reaction mixture was heated at 80° C. for 15 h, allowed to cool and passed through a SCX-2 cartridge (1 g), eluting with 0.5M NH3 in MeOH. The crude material was concentrated in vacuo and purified by reverse phase preparative HPLC-MS to afford 5-methyl-3-[4-(2-methylamino-ethoxy)-phenyl]-2H-isoquinolin-1-one as a white solid (4 mg, 12%).


AnalpH2_MeOH_QC: Rt 5.43 min; m/z 309 [M+1]+.


Scheme A, Step C (Protocol 1): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 5 (via BOC deprotection)
N-Methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-piperidin-4-ylmethyl-benzamide (IQ-093)



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To 4-({methyl-[4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoyl]-amino}-methyl)-piperidine-1-carboxylic acid tert-butyl ester (170 mg, 0.35 mmol) in CH2Cl2 (5 mL) was added 4M HCl/dioxane (2 mL) and the reaction mixture stirred at RT for 4 h. The solvent was removed in vacuo and the crude product purified by reverse phase preparative HPLC-MS to obtain N-methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-piperidin-4-ylmethyl-benzamide as a pale orange solid (44 mg, 33%).



1H NMR (400 MHz, DMSO-d6): δ8.09 (d, J=8 Hz, 1H), 7.89 (d, J=8 Hz, 2H), 7.57 (d, J=7 Hz, 1H), 7.50 (br d, J=8 Hz, 1H), 7.45 (br d, J=8 Hz, 1H), 6.92 (s, 1H), 3.35 (d, J=7 Hz, 1H), 3.15 (d, J=7 Hz, 1H), 2.97 (s, 2H), 2.92 (s, 3H), 2.84-2.82 (m, 1H), 2.51 (s, 3H), 2.46-2.36 (m, 1H), 1.84 (s, 0.5H), 1.77 (s, 0.5H), 1.61 (d, J=10 Hz, 1H), 1.42 (d, J=10 Hz, 1H), 1.11-1.08 (m, 1H), 0.70-0.68 (m, 1H).


AnalpH2_MeOH_QC(Sunfire1): Rt 4.49 min; m/z 390 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 13







2H-isoquinolin-1-one Formula 5










Compound
Code
Analytical Data
Mass, % Yield, State







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IQ-094
AnalpH2_MeOH_QC (Sunfire1): Rt 4.82 min; m/z 410 [M + 1]+
66 mg, 60%, white solid







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IQ-070
AnalpH2_MeOH_QC(1): Rt 5.19 min; m/z 338 [M + 1]+
63 mg, 94%, pale pink solid







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IQ-067
AnalpH2_MeOH_QC(1): Rt 5.57 min; m/z 348 [M + 1]+
14 mg, 12%, orange solid







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IQ-073
AnalpH2_MeOH_QC(1): Rt 5.32 min; m/z 348 [M + 1]+
7 mg, 28%, white solid







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IQ-090
AnalpH2_MeOH_QC(1): Rt 3.96 min; m/z 360 [M + 1]+
48 mg, 33%, pale yellow solid







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IQ-062
AnalpH2_MeOH_QC(1): Rt 4.23 min; m/z 348 [M + 1]+
211 mg, 100%, pale orange solid







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IQ-051-1
AnalpH2_MeOH_QC: Rt 5.65 min; m/z 348 [M + 1]+
108 mg, 77%, cream solid







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IQ-051-2
AnalpH2_MeOH_QC(1): Rt 5.47 min; m/z 348 [M + 1]+
10.2 mg, 35% recovery, off-white solid; obtained via Chiral_Method_2







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IQ-051-3
AnalpH2_MeOH_QC(1): Rt 5.47 min; m/z 348 [M + 1]+
8.3 mg, 29% recovery, off-white solid; obtained via Chiral_Method_2







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IQ-084-3
AnalpH2_MeOH_QC: Rt 5.61 min; m/z 348 [M + 1]+
34 mg, 42%, cream solid







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IQ-084-1
AnalpH2_MeOH_QC (Sunfire1): Rt 4.67 min; m/z 348 [M + 1]+
4.5 mg, 13% recovery, white solid; obtained via Chiral_Method_1







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IQ-084-2
AnalpH2_MeOH_QC (Sunfire1): Rt 4.66 min; m/z 348 [M + 1]+
3.5 mg, 11% recovery, white solid; obtained via Chiral_Method_1







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IQ-063
AnalpH2_MeOH_QC(1): Rt 5.56 min; m/z 362 [M + 1]+
87 mg, 55%, cream solid







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IQ-059
AnalpH2_MeOH_QC(1): Rt 5.51 min; m/z 352 [M + 1]+
112 mg, 100%, pale orange solid







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IQ-082
AnalpH2_MeOH_QC: Rt 5.36 min; m/z 334 [M + 1]+
107 mg, 38%, white solid







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IQ-083
AnalpH2_MeOH_QC: Rt 5.87 min; m/z 354 [M + 1]+
115 mg, 97%, yellow yellow solid







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IQ-086
AnalpH2_MeOH_QC: Rt 4.77 min; m/z 346 [M + 1]+
22 mg, 72%, white solid







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IQ-049
AnalpH2_MeOH_QC: Rt 5.91 min; m/z 360 [M + 1]+
25 mg, 96%, off- white solid







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IQ-029
AnalpH2_MeOH_QC: Rt 5.00 min; m/z 279 [M + 1]+
9 mg, 24%, off- white solid







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IQ-150
AnalpH2_MeOH_QC(1): Rt 5.83 min; m/z 400 [M + 1]+
367 mg, 56%, cream solid







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IQ-158
AnalpH2_MeOH_QC: Rt 5.62 min; m/z 356 [M + 1]+
22 mg, 65% white solid







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IQ-081
AnalpH2_MeOH_QC(1): Rt 5.71 min; m/z 348 [M + 1]+
29 mg, 48%, pale peach solid







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IQ-028-1
AnalpH2_MeOH_QC (Sunfire1): Rt 4.29 min; m/z 321 [M + 1]+
212 mg, 45%, orange solid 1H NMR (400 MHz, DMSO-d6): δ11.42 (br s, 1H), 8.58 (d, J = 2.3 Hz, 1H), 8.05 (d, J = 7.8 Hz, 1H), 7.97 (dd, J = 8.8, 1.0 Hz, 1H), 7.53 (d, J = 6.8 Hz, 1H), 7.32 (t, J = 7.6 Hz, 1H), 6.9 (d, J = 8.8 Hz, 1H), 6.77 (s, 1H), 3.53 (t, J = 5.1 Hz, 4H), 2.81 (t, J = 5.1 Hz, 4H), 2.54 (s, 3H).







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IQ-027
AnalpH2_MeOH_QC (Sunfire1): Rt 4.73 min; m/z 341 [M + 1]+
460 mg, 66%, pale yellow solid 1H NMR (400 MHz, DMSO-d6): δ8.54 (d, J = 2.2 Hz, 1H), 8.16 (d, J = 7.8 Hz, 1H), 7.93 (dd, J = 9.1, 2.5 Hz, 1H), 7.85 (dd, J = 7.6 Hz, 1.0 1H), 7.43 (t, J = 7.8 Hz, 1H), 6.90 (d, J = 9.1 Hz, 1H), 6.83 (s, 1H), 3.54-3.52 (m, 4H), 2.80-2.78 (m, 4H).







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IQ-024
AnalpH2_MeOH_QC: Rt 5.74 min; m/z 359 [M + 1]+
82 mg, 67%, pale orange solid







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IQ-023
AnalpH2_MeOH_QC: Rt 5.41 min; m/z 339 [M + 1]+
51 mg, 97%, yellow solid







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IQ-016
AnalpH2_MeOH_QC: Rt 5.35 min; m/z 320 [M + 1]+
59 mg, 41%, beige solid







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IQ-014
AnalpH2_MeOH_QC: Rt 5.65 min; m/z 340 [M + 1]+
20 mg, 10%, yellow yellow solid







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IQ-115
AnalpH2_MeOH_QC (Sunfire1): RT 4.81 min; m/z 426 [M + 1]+.
60 mg, 32%, cream solid







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IQ-116
AnalpH2_MeOH_QC (Sunfire1): RT 5.13 min; m/z 446 [M + 1]+.
36 mg, 21%, pale orange solid







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IQ-124
AnalpH2_MeOH_QC: RT 5.79 min; m/z 404 [M + 1]+.
146 mg, 89%, yellow yellow solid







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IQ-123
AnalpH2_MeOH_QC: RT 5.50 min; m/z 384 [M + 1]+.
54 mg, 59%, yellow yellow solid







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IQ-159
AnalpH2_MeOH_QC: Rt 6.08 min; m/z 360 [M + 1]+
4.4 mg, 17% pale cream solid









Scheme A, Step C (Protocol 2): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 5 (via TBDMS Deprotection)
3-{6-[4-(2-Hydroxy-ethyl)-piperazin-1-yl]-pyridin-3-yl}-5-methyl-2H-isoquinolin-1-one (IQ-011)



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To a solution of 3-(6-{4-[2-(tert-butyl-dimethyl-silanyloxy)-ethyl]-piperazin-1-yl}-pyridin-3-yl)-5-methyl-2H-isoquinolin-1-one (214 mg, 0.45 mmol) in THF (1.5 mL) at 5° C. was added 1M TBAF/THF (0.58 mL, 0.58 mmol) dropwise. The reaction mixture was allowed to warm to RT and stirred for 1 h. The reaction mixture was diluted with EtOAc (10 mL) and washed with water and brine. The organic layer was concentrated in vacuo and purified by reverse phase preparative HPLC-MS to obtain 3-{6-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-pyridin-3-yl}-5-methyl-2H-isoquinolin-1-one as a yellow solid (27 mg, 16%).



1H NMR (400 MHz, DMSO-d6): δ11.61-11.29 (br s, 1H), 8.59 (d, J=2.5 Hz, 1H), 8.05 (d, J=8 Hz, 1H), 7.98 (dd, J=9, 2.5 Hz, 1H), 7.53 (d, J=8 Hz, 1H), 7.32 (t, J=8 Hz, 1H), 6.92 (d, J=9 Hz, 1H), 6.77 (s, 1H), 4.43 (t, J=5 Hz, 1H), 3.59-3.54 (m, 6H), 2.54 (s, 3H), 2.54-2.52 (m, 4H), 2.44 (t, J=5 Hz, 2H).


AnalpH2_MeOH_QC: Rt 5.04 min; m/z 365 [M+1]+.


The following 2H-isoquinolin-1-one of formula 5 derivatives are prepared using analogous procedures.









TABLE 14







2H-isoquinolin-1-one derivatives of Formula 5













Mass, % Yield,


Compound
Code
Analytical Data
State







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IQ-012
AnalpH2_MeOH_QC: Rt 5.40 min; m/z 385 [M + 1]+
21 mg, 12%, pale yellow solid







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IQ-219
AnalpH2_MeOH_QC(1): Rt 5.08 min; m/z 326 [M + 1]+
65 mg, 13%, white solid









Scheme A, Step C (Protocol 3): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 5 (via TBDPS Deprotection)
3-[4-(4-Hydroxy-piperidin-1-ylmethyl)-phenyl]-5-methyl-2H-isoquinolin-1-one (IQ-074)



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To a stirred solution of 3-{4-[4-(tert-butyl-diphenyl-silanyloxy)-piperidin-1-ylmethyl]-phenyl}-5-methyl-2H-isoquinolin-1-one (213 mg, 0.36 mmol) in CH2Cl2 (2 mL) was added 1.25M methanolic HCl (1 mL) and the reaction stirred at RT for 48 h. The reaction mixture was concentrated in vacuo and the crude residue purified by reverse phase preparative HPLC-MS to afford 3-[4-(4-hydroxy-piperidin-1-ylmethyl)-phenyl]-5-methyl-2H-isoquinolin-1-one as a pale yellow solid (80 mg, 64%).


AnalpH2_MeOH_QC(1): Rt 5.03 min; m/z 349 [M+1]+.



1H NMR (400 MHz, DMSO-d6): δ11.63-11.35 (br s, 1H), 8.15 (s, 0.8H) 8.07 (d, J=7 Hz, 1H), 7.78 (d, J=8 Hz, 2H), 7.56 (d with fine coupling, J=7 Hz, 1H), 7.41 (d, J=8 Hz, 2H), 7.37 (t, J=8 Hz, 1H), 6.85 (s, 1H), 4.66-4.52 (br s, 1H), 3.52 (s, 2H), 3.50-3.45 (m, 1H), 2.70-2.67 (m, 2H), 2.56 (s, 3H), 2.11-2.06 (m, 2H), 1.74-1.70 (m, 2H), 1.45-1.36 (m, 2H).


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 15







2H-isoquinolin-1-one derivatives of Formula 5













Mass, % Yield,


Compound
Code
Analytical Data
State







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IQ-120
AnalpH2_MeOH_ QC(1): Rt 7.31 min; m/z 373 [M + 1]+
39 mg, 60%, white solid







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IQ-167
AnalpH2_MeOH_ QC(1): Rt 5.04 min; m/z 321 [M + 1]+
70 mg, 50%, off- white solid







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IQ-169
AnalpH2_MeOH_ QC(1): Rt 5.11 min; m/z 339 [M + 1]+
3.5 mg, 15%, off- white solid







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IQ-172
AnalpH2_MeOH_ QC(1): Rt 5.09 min; m/z 335 [M + 1]+
17 mg, 49%, off- white solid









Scheme A, Step C (Protocol 4): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 5 (via Tosyl Deprotection) (IQ-075)



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To a solution of 5-methyl-3-(4-{1-methyl-1-[4-(toluene-4-sulfonyl)-piperazin-1-yl]-ethyl}-phenyl)-2H-isoquinolin-1-one (33 mg, 0.064 mmol) in HBr (33% w/w in acetic acid) (0.25 mL) was added 4-hydroxybenzoic acid (27 mg, 0.194 mmol) and the reaction stirred for 16 h at RT afterwhich time the reaction was concentrated in vacuo and the crude residue purified by preperative HPLC to afford 5-Methyl-3-[(4-(1-methyl-1-piperazin-1-yl]-ethyl)-phenyl]-2H-isoquinolin-1-one as a pale orange solid (1.06 mg, 4.5%).


AnalpH2_MeOH_QC(1): RT 6.01 min; m/z 362 [M+1]+.


Scheme A, Step D (Protocol 1): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 6 (via acylation)
3-[4-(4-Acetyl-piperazin-1-ylmethyl)-phenyl]-5-methyl-2H-isoquinolin-1-one (IQ-055)



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To a stirred solution of acetic acid (0.005 mL, 0.068 mmol) in CH2Cl2 (5 mL) was added TBTU (22 mg, 0.068 mmol) and N,N-diisopropylethylamine (0.036 mL, 0.20 mmol) and the reaction stirred for 10 min at RT. 5-Methyl-3-(4-piperazin-1-ylmethylphenyl)-2H-isoquinolin-1-one (23 mg, 0.068 mmol) was then added and the reaction stirred for 2 h at RT. The reaction mixture was concentrated in vacuo and purified by reverse phase preparative HPLC-MS to afford 3-[4-(4-acetyl-piperazine-1-ylmethyl)-phenyl]-5-methyl-2H-isoquinolin-1-one as a white solid (2 mg, 9%).



1H NMR (400 MHz, DMSO-d6): δ 11.70-11.38 (br s, 1H), 8.08 (d, J=8 Hz, 1H), 7.80 (d, J=8 Hz, 2H), 7.56 (d, J=8 Hz, 1H), 7.44 (d, J=8 Hz, 2H), 7.37 (t, J=8 Hz, 1H), 6.92 (s, 1H), 3.57 (s, 2H), 3.46-3.42, (m, 4H), 2.56 (s, 3H), 2.44-2.39 (m, 2H), 2.35-2.32 (m, 2H), 1.99 (3H, s).


AnalpH2_MeOH_QC(Sunfire): Rt 4.38 min; m/z 376 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 16







2H-isoquinolin-1-one derivatives of Formula 6












Analytical
Mass, % Yield,


Compound
Code
Data
State







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IQ-017
AnalpH2_ MeOH_QC: Rt 7.76 min; m/z 362 [M + 1]+
18 mg, 36%, beige solid







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IQ-015
AnalpH2_ MeOH_QC: Rt 8.16 min; m/z 382 [M + 1]+
11 mg, 43%, yellow solid







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IQ-045
AnalpH2_ MeOH_QC Rt 5.62 min m/z 396 [M + 1]+.
17 mg, 42%, white solid









Scheme A, Step D (Protocol 2): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 6 (via acylation)
3-[2-(4-Cyclopropanecarbonyl-piperazin-1-yl)-pyrimidin-5-yl]-5-methyl-2H-isoquinolin-1-one (IQ-157)



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Cyclopropylcarbonyl chloride (9 μL, 2.28 mmol) was added to a stirred solution of 5-methyl-3-(2-piperazin-1-yl-pyrimidin-5-yl)-2H-isoquinolin-1-one (36 mg, 0.11 mmol) and N,N-diisopropylethylamine (23 μL, 0.132 mmol) in CH2Cl2 (2 mL) at −20° C. and allowed to stir for 10 min. The reaction mixture was concentrated in vacuo. The crude residue was purified by reverse phase preparative HPLC-MS to afford 3-[2-(4-cyclopropanecarbonyl-piperazin-1-yl)-pyrimidin-5-yl]-5-methyl-2H-isoquinolin-1-one as a white solid (19 mg, 45%).



1H NMR (400 MHz, DMSO-d6): δ11.82-11.11 (br s, 1H), 8.85 (s, 2H), 8.05 (d, J=8 Hz, 1H), 7.55 (d, J=8 Hz, 1H), 7.35 (t, J=8 Hz, 1H), 6.84 (s, 1H), 3.96-3.76 (br m, 6H), 3.64-3.54 (br s, 2H), 2.54 (s, 3H), 2.08-2.00 (m, 1H), 0.82-0.70 (m, 4H).


AnalpH2_MeOH_QC(1): Rt 8.05 min; m/z 390 [M+1]+.


Scheme A, Step D (Protocol 3): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 6 (urea formation) (IQ-068)



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To a stirred solution of 5-methyl-3-(4-piperazin-1-ylmethyl-phenyl)-2H-isoquinolin-1-one (40 mg, 0.12 mmol) in CH2Cl2 (0.5 mL) was added tert-butyl isocyanate (0.014 mL, 0.12 mmol) and the reaction mixture stirred at RT for 1 h after which time the solvent was removed in vacuo and the crude residue purified by reverse phase preparative HPLC-MS to afford 4-[4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzyl]-piperazine-1-carboxylic acid tert-butylamide as a white solid (28 mg, 54%).



1H NMR (400 MHz, DMSO-d6): δ 11.50-11.12 (brs, 1H), 7.83 (d, J=8 Hz, 1H), 7.55 (d, J=8 Hz, 2H), 7.32 (d, J=8 Hz, 1H), 7.19 (d, J=8 Hz, 2H), 7.13 (t, J=8 Hz, 1H), 6.62 (s, 1H), 5.49 (s, 1H), 3.30 (s, 2H), 3.03-3.01, (m, 4H) 2.32 (s, 3H), 2.10-2.08 (m, 4H), 1.01 (s, 9H).


AnalpH2_MeOH_QC(1): Rt 5.83 min; m/z 433 [M+1]+.


Synthesis of 3-(1-Oxy-pyridin-3-yl]-2H-isoquinolin-1-one Derivatives 27



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Scheme H, Step P: N-Oxidation of 3-(pyridinyl)-2H-isoquinolin-1-one Derivatives 27
5-Chloro-3-[6-(4-methyl-piperazin-1-yl)-1-oxy-pyridin-3-yl]-2H-isoquinolin-1-one (IQ-002-2)



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MCPBA (41 mg, 0.184 mmol) was added to a solution of 5-chloro-3-[6-(4-methyl-piperazin-1-yl)-pyridin-3-yl]-2H-isoquinolin-1-one (58 mg, 0.164 mmol) in CH2Cl2 (7 mL) at −78° C. and allowed to warm to RT and stirred at this temperature for a further 40 minutes. The reaction mixture was quenched with saturated, aqueous NaHCO3 (2 mL) and extracted with CH2Cl2 followed by EtOAc. The combined organic layer was concentrated in vacuo and passed through an SCX-2 cartridge (5 g), eluting with 10% NH3/MeOH. The desired fractions were concentrated in vacuo and the crude material purified by reverse phase preparative HPLC-MS to obtain 5-chloro-3-[6-(4-methyl-piperazin-1-yl)-1-oxy-pyridin-3-yl]-2H-isoquinolin-1-one as a white solid (19 mg, 33%).



1H NMR (400 MHz, DMSO-d6): δ12.30-12.00 (brs, 1H), 8.59 (d, J=8 Hz, 1H), 8.31 (s, 0.6H), 8.17 (d, J=9 Hz, 1H), 8.01 (dd, J=9, 3 Hz, 1H), 7.84 (d, J=8 Hz, 1H), 7.43 (t, J=8 Hz, 1H), 7.03 (d, J=9 Hz, 1H), 6.84 (s, 1H), 4.29-4.27 (m, 2H), 3.59-3.49 (m, 4H), 3.35-3.31 (m, 2H), 3.25 (s, 3H).


AnalpH9_MeOH_QC: Rt 7.20 min; m/z 371 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 17







2H-isoquinolin-1-one derivatives 27













Mass, % Yield,


Compound
Code
Analytical Data
State







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IQ-028-2
AnalpH9_MeOH_QC (Sunfire1): Rt 5.86 min; m/z 337 [M + 1]+
48 mg, 57%, brown solid









General Procedure for Synthesis of 2H-isoquinolin-1-one derivatives of Formula 34



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Scheme I, Step Q Synthesis of 3-(4-Bromo-phenyl)-5-methyl-2H-isoquinolin-1-one



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To N,N-diisopropylamine (2.54 mL, 18 mmol) in anhydrous THF (15 mL), under N2 at −78° C. was added n-BuLi dropwise (2.5M in n-hexanes, 7.2 mL, 18 mmol) and the reaction mixture maintained at this temperature for 30 minutes. A solution of N,N-diethyl-2,3-dimethyl-benzamide (1.23 g, 6 mmol) in anhydrous THF (15 mL) was added dropwise to give a deep red solution. After 20 minutes at −78° C., 4-bromobenzonitrile (1.09 g, 6 mmol) in anhydrous THF (15 mL) was added dropwise and the reaction mixture allowed to stir at this temperature for 2.5 h. The reaction mixture was quenched by adding dropwise onto ice, upon which a pale yellow solid precipitated out. The solid was triturated with iso-hexane/EtOAc (2:1), filtered and dried in vacuo to afford 3-(4-bromo-phenyl)-5-methyl-2H-isoquinolin-1-one as a pale yellow solid (1.1 g, 58%).


AnalpH2_MeOH—4 min(1): Rt 3.25 min; m/z 314 [M+1]+.


Scheme I, Step R: Synthesis of 4-(5-Methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzonitrile



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3-(4-Bromo-phenyl)-5-methyl-2H-isoquinolin-1-one (200 mg, 0.64 mmol), zinc cyanide (90 mg, 0.76 mmol) and tetrakis(triphenylphosphine)palladium(0) (74 mg, 0.064 mmol) were stirred in DMF (2.1 mL) and degassed with N2. The reaction mixtures were irradiated using a microwave (300 W, 180° C., 30 min). The reaction mixtures were combined and the resulting precipitate was filtered, washed with DMF and water and dried to give 4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzonitrile as a yellowish solid (718 mg, 79%) which was used in the next step without further purification.


AnalpH2_MeOH—4 min(1): Rt 2.83 min; m/z 261 [M+1]+.


Scheme I, Step S: Synthesis of 4-(5-Methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoic acid



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To 4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzonitrile (100 mg, 0.38 mmol) was added 2M NaOH (1.5 mL) and the reaction mixture irradiated using a microwave (300 W, 130° C., 20 min). The reaction mixtures was diluted with water and adjusted to pH2 with 2M HCl whereupon a pale yellow solid precipitated out of solution. The solid was filtered, washed with water and dried. The solid was dissolved in DMF and passed through a Si-thiol cartridge to remove any residual palladium, eluting with DMF. The eluent was removed in vacuo to give 4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoic acid as a pale yellow solid (120 mg, 63%).


AnalpH9_MeOH—4 min(1): Rt 2.24 min; m/z 280 [M+1]+.


Scheme I, Step T: Synthesis of 3-Benzamide-5-Methyl-2H-Isoquinolin-1-one Derivatives of formula 33
N-Methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(tetrahydro-pyran-4-ylmethyl)-benzamide (IQ-097)



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To 4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoic acid (35 mg, 0.125 mmol), TBTU (40 mg, 0.125 mmol) was added 0.36M N,N-diisopropylethylamine/CH2Cl2 (0.35 mL, 0.125 mmol) and anhydrous DMF (0.9 mL). The reaction mixture was stirred at RT for 45 min after which time methyl-(tetrahydro-pyran-4-ylmethyl)-amine (19 mg, 0.15 mmol) in anhydrous DMF (0.45 mL) was added and the reaction mixture was stirred at RT overnight. The reaction mixture was passed through a Si—NH2 cartridge (1 g), eluting with DMF (2× column volumes), MeOH (2× column volumes) and the solvent removed in vacuo and the crude product purified by reverse phase preparative HPLC-MS to obtain N-methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(tetrahydro-pyran-4-ylmethyl)-benzamide as a yellow foam (24 mg, 48%).



1H NMR (400 MHz, DMSO-d6): δ11.69-11.54 (br s, 1H), 8.09 (d, J=7 Hz, 1H), 7.89 (d, J=8 Hz, 2H), 7.58 (d, J=7 Hz, 1H), 7.52 (br d, J=8 Hz, 1H), 7.46 (br d, J=8 Hz, 1H), 7.40 (t, J=7 Hz, 1H), 6.92 (s, 1H), 3.90 (br d, J=11 Hz, 1H), 3.75 (br d, J=11 Hz, 1H), 3.40-3.17 (m, 4H), 2.99 (br s, 1H), 2.94 (br s, 2H), 2.58 (s, 3H), 2.08-1.82 (br m, 1H), 1.63 (br d, J=12 Hz, 1H), 1.44 (br d, J=12 Hz, 1H), 1.32-1.25 (m, 1H), 0.95-0.80 (m, 1H).


AnalpH2_MeOH_QC(1): Rt 7.69 min; m/z 391 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures









TABLE 18







2H-isoquinolin-1-one derivatives of Formula 33













Mass, % Yield,


Compound
Code
Analytical Data
State







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IQ-107
AnalpH2_MeOH_ QC(1): Rt 7.44 min; m/z 349 [M + 1]+
12 mg, 51%, white solid







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IQ-146
AnalpH2_MeOH_ QC(1): Rt 5.27 min; m/z 376 [M + 1]+
16 mg, 35%, off-white solid







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IQ-098
AnalpH2_MeOH_ QC(1): Rt 5.51 min; m/z 416 [M + 1]+
15 mg, 29%, off-white solid







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IQ-153
AnalpH2_ MeOH_4 min: Rt 3.15 min; m/z 476 [M + 1]+
21 mg, 100%, beige solid









Scheme I, Step U: Synthesis of 3-Benzamide-5-Methyl-2H-Isoquinolin-1-one Derivatives of formula 34
N-Methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-piperidin-4-yl-benzamide (IQ-096)



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To 4-{methyl-[4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoyl]-amino}-piperidine-1-carboxylic acid tert-butyl ester (21 mg, 0.044 mmol) was added 2:1 CH2Cl2/TFA (1 mL) and the reaction mixture stirred at RT for 1 h. The solvent was removed in vacuo, re-dissolved in MeOH and passed through an SCX-2 cartridge (1 g). The column was washed with MeOH (4× column volumes), the desired product eluted from the cartridge with 0.5M NH3/MeOH (4× column volumes) and concentrated in vacuo. The crude product was purified by reverse phase preparative HPLC-MS to obtain N-methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-piperidin-4-yl-benzamide as a white solid (2.4 mg, 14%).


AnalpH2_MeOH_QC(1): Rt 5.10 min; m/z 376 [M+1]+.


Synthesis of 2H-isoquinolin-1-one derivatives of Formula 4 & 5 (via Route 2)



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Scheme J, Step V: Synthesis of Phenyl Acrylic Acid Derivatives of Formula 36
(E)-3-(4-Fluoro-2-methylphenyl)acrylic acid



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A stirred solution of 4-fluoro-2-methyl-benzaldehyde (20 g, 144.9 mmol) and malonic acid (30.1 g, 289.8 mmol) in pyridine (100 mL) was heated to 50° C. Piperidine (10 mL) was added and the reaction mixture was heated at 70° C. for 18 h. The reaction mixture was cooled RT and poured into chilled aqueous 1N HCl solution (1500 mL), the resulting precipitate was filtered and washed with petroleum ether 60-80 and dried in vacuo to obtain (E)-3-(4-fluoro-2-methylphenyl)acrylic acid (18 g, 69%) as an off white solid.



1H NMR (400 MHz, CDCl3): δ8.00 (d, J=16 Hz, 1H), 7.60-7.55 (m, 1H), 6.96-6.90 (m, 2H), 6.32 (d, J=16 Hz, 1H), 2.44 (s, 3H).


AnalpH2_MeCN_FA—7 min(XTERRA1.m): Rt 3.34 min; m/z 181 [M+1]+.


The following phenyl acrylic acid derivatives of formula 36 are prepared using analogous procedures.









TABLE 19







Phenyl acrylic acid Derivatives 36













Mass,





%




Analytical
Yield,


Compound
Reference
Data
State







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Reported as commer- cially available
AnalpH2_ MeCN_ FA_7 min (XTERR A1.m): Rt 3.16 min; m/z 185 [M + 1]+.
25 g, 64%, off- white solid







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Reported as commer- cially available
AnalpH2_ MeOH_ 4 min(3): Rt 2.63 min; m/z not observed
6.3 g, 97%, white solid









Scheme J, Step W: Synthesis of Phenyl Propanoic Derivatives 37
3-(4-Fluoro-2-methylphenyl)propanoic acid



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To a solution of (E)-3-(4-fluoro-2-methylphenyl)acrylic acid (13 g, 72.22 mmol) in EtOH (250 mL) was added PtO2 (250 mg) and then hydrogenated at 30 psi for 3 h. The reaction mixture was filtered on a Celite® pad, washed with MeOH (100 mL), and the filtrate was concentrated, washed with diethyl ether (20 mL), n-pentane (50 mL) and dried in vacuo to give 3-(4-fluoro-2-methylphenyl)propanoic acid as an off-white solid (10 g, 77%).



1H NMR (400 MHz, CDCl3): δ 7.13-7.05 (m, 1H), 6.90-6.79 (m, 2H), 2.95-2.85 (2H, m), 2.65-2.55 (2H, m), 2.31 (s, 3H)


AnalpH2_MeCN_FA—7 min(XTERRA1.m): Rt 3.33 min; m/z 181 [M−1]−.


The following phenyl propanoic derivatives 37 are prepared using analogous procedures.









TABLE 20







Phenyl Propanoic Derivatives 37













Mass,


Compound
Reference
Analytical Data
% Yield, State







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Reported as commercially available
AnalpH2_MeCN_ FA_7 min (XTERRA1.m): Rt 3.19 min; m/z 185 [M − 1]−.
8 g, 73%, off- white solid







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Reported as commercially available
AnalpH2_MeOH_ 4 min(3): Rt 2.58 min; m/z not observed
6.37 g, 100%, white solid









Scheme J, Step X: Indanone Synthesis
6-Fluoro-4-methyl-2,3-dihydro-1H-inden-1-one



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To a solution of 3-(4-fluoro-2-methylphenyl)propanoic acid (12 g, 65.93 mmol) in CH2Cl2 (200 mL) was added oxalyl chloride (11.3 mL, 131.7 mmol) and stirred at RT for 16 h. The reaction mixture was concentrated in vacuo and re-dissolved in CH2Cl2 (150 mL) and added to a suspension of AlCl3 (11.4 g, 85.7 mmol) in CH2Cl2 (150 mL) at 0° C. The reaction mixture was heated at 50° C. for 3 h and allowed to stir at RT for 16 h. The reaction mixture poured into ice water (150 mL), extracted with CH2Cl2 (2×100 mL), the organic extract was washed with 1N NaOH solution (2×50 mL), brine solution (50 mL), dried over Na2SO4 and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with 10% EtOAc/petroleum ether 60-80 to afford 6-fluoro-4-methyl-2,3-dihydro-1H-inden-1-one as an off white solid (7 g, 70%).



1H NMR (400 MHz, CDCl3): δ 7.28-7.25 (m, 1H), 7.18-7.12 (m, 1H), 3.03-2.96 (m, 2H), 2.78-2.73 (m, 2H), 2.35 (s, 3H).


AnalpH2_MeCN_TFA—4 min(1): Rt 1.89 min; m/z 165 [M+1]+.


The following indanone derivatives 38 are prepared using analogous procedures.









TABLE 21







Indanone Derivatives 38













Mass,




Analytical
% Yield,


Compound
Reference
Data
State







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Commercially available
N/A
N/A







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Reported as commercially available
AnalpH2_ MeCN_ TFA_4 min(1): Rt 1.80 min; m/z 169 [M + 1]+.
5.2 g, 57%, off-white solid







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Commercially available
N/A
N/A







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Reported as commercially available
AnalpH2_ MeOH_ 4 min(3): Rt 2.23 min; m/z 165[M + 1]+
5.62 g, 98%, off-white solid









Scheme J, Step Y: Synthesis 2-(hydroxyimino-2,3-dihydro-1H-inden-1-one Derivatives 39
6-Fluoro-2-(hydroxyimino)-4-methyl-2,3-dihydro-1H-inden-1-one



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To a stirred solution of 6-fluoro-4-methyl-2,3-dihydro-1H-inden-1-one (1 g, 6.09 mmol) in a mixture of diethyl ether (10 mL) and concentrated HCl (10 mL) was added isopentyl nitrite (0.73 mL, 5.47 mmol) and stirred at RT for 3 h. The precipitated solid was collected by filtration and washed with MeOH to obtain 6-fluoro-2-(hydroxyimino)-4-methyl-2,3-dihydro-1H-inden-1-one as a brown solid (800 mg, 68%).



1H NMR (400 MHz, DMSO-d6): δ12.73 (s, 1H), 7.52-7.45 (m, 1H), 7.32-7.29 (m, 1H), 3.67 (s, 2H), 2.35 (s, 3H).


AnalpH2_MeCN_FA—7 min(XTERRA1.m): Rt 3.04 min; m/z 194 [M+1]+.


The following 2-(hydroxyimino-2,3-dihydro-1H-inden-1-one derivatives 39 are prepared using analogous procedures.









TABLE 22







2-(hydroxyimino-2,3-dihydro-1H-inden-1-one Derivatives of formula 39













Mass,



Refer-

% Yield,


Compound
ence
Analytical Data
State







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AnalpH9_MeCN_ AB_10 min (Develosil): Rt 2.85 min; m/z 176 [M + 1]+
5 g, 43%, pale yellow solid.







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AnalpH2_MeCN_ FA_7 min (XTERRA1.m): Rt 2.93 min; m/z 198 [M + 1]+
1 g, 57% brown solid







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AnalpH2_MeOH_ 4 min(1): Rt 2.08 min; m/z 198 [M + 1]+
4.27 g, 72%, beige solid







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AnalpH2_MeOH_ 4 min(3): Rt 2.17 min; m/z 194 [M + 1]+
3.84 g, 58%, pale brown solid









Scheme J, Step Z: Synthesis of 3-chloro-isoquinolin-1(2H)-one derivatives of formula 40
3-Chloro-7-fluoro-5-methylisoquinolin-1(2H)-one



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To a solution of 6-fluoro-2-(hydroxyimino)-4-methyl-2,3-dihydro-1H-inden-1-one (800 mg, 4.12 mmol) in anhydrous CCl4 (100 mL) was added PCl5 (1.28 g, 6.18 mmol) and stirred at RT for 16 h. The reaction mixture was concentrated in vacuo and the residue dissolved in anhydrous 1,4-dioxane (100 mL), cooled 0° C., the solution was saturated with HCl gas and allowed to stir RT for 16 h. The reaction mixture was heated at 60° C. for 2 h, cooled to RT and diluted with EtOAc (50 mL), washed with water (25 mL), saturated NaHCO3 solution (25 mL), brine solution (25 mL), dried (Na2SO4), filtered and concentrated in vacuo. The crude material was washed with diethyl ether (10 mL), n-pentane (10 mL) and was dried in vacuo to obtain 3-chloro-7-fluoro-5-methylisoquinolin-1(2H)-one as a pale yellow solid (550 mg, 68%).



1H NMR (400 MHz, DMSO-d6): δ12.55-12.40 (br s, 1H), 7.70-7.63 (m, 1H), 7.55-7.49 (m, 1H), 6.84-6.70 (br s, 1H), 2.48 (s, 3H).


AnalpH2_MeOH—4 min(1): Rt 2.74 min; m/z 212 [M+1]+.


The following 3-chloro-isoquinolin-1(2H)-one derivatives 40 are prepared using analogous procedures.









TABLE 23







3-Chloro-isoquinolin-1(2H)-one Derivatives of formula 40













Mass,





% Yield,


Compound
Reference
Analytical Data
State







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Compound reported by Krämer et al., 1969
AnalpH2_MeOH_ 4 min: Rt 2.67 min(1); m/z 193 [M + 1]+
1.1 g, 20%, white solid







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Novel
AnalpH2_MeOH_ 4 min(1): Rt 2.78 min; m/z 216 [M + 1]+
650 mg, 62%, off white solid







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Novel
AnalpH2_MeOH_ 4 min(1): Rt 2.51 min; m/z 215 [M +1]+
150 mg, 28%, off- white solid







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AnalpH2_MeOH_ 4 min(3): Rt 2.46 min; m/z 212 [M + 1]+
2.43 g, 58%, pale yellow solid









Synthesis of Boronic Acid/Ester Intermediates 43 (required for Step AA, Scheme J)



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Scheme JJ: Synthesis of an Example of Amine of Formula 9
1-[2-(tert-Butyl-diphenyl-silanyloxy)-ethyl]-piperazine



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To 2-piperazin-1-yl-ethanol (2 g, 15.36 mmol) in CH2Cl2 (70 mL) and pyridine (1.85 mL, 23.04 mmol) was added DMAP (188 mg, 1.53 mmol) and TBDPS chloride (3.37 mL, 18.44 mmol) and the reaction mixture stirred at RT for 18 h. The reaction mixture was concentrated in vacuo and the crude material was purified by silica gel column chromatography, eluting with CH2Cl2 and increasing the polarity to 10% MeOH/CH2Cl2 to obtain 1-[2-(tert-butyl-diphenyl-silanyloxy)-ethyl]-piperazine as a colourless oil (1.1 g, 21%).


AnalpH2_MeOH—4 min(3): Rt 2.48 min; m/z 369 [M+1]+.


Scheme JJ, Step AC (Protocol 1): Synthesis of Aryl Boronic Acid or Boronic Ester Derivatives of Formula 43 (Via Amide Coupling)
2-Fluoro-4-(morpholine-4-carbonyl)-boronic acid



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To 4-carboxy-2-fluoro-benzene boronic acid (150 mg, 0.82 mmol), TBTU (262 mg, 0.82 mmol) in anhydrous DMF (8 mL), was added 0.36M N,N-diisopropylethylamine in anhydrous CH2Cl2 (2.3 mL, 0.82 mmol) and the reaction mixture stirred at RT for 45 min. Morpholine (85 mg, 0.99 mmol) in anhydrous DMF (1 mL) was added and the reaction mixture stirred for 18 hr at RT. The reaction mixture was concentrated in vacuo and the crude material was purified by reverse phase preparative HPLC-MS to afford 2-fluoro-4-(morpholine-4-carbonyl)-boronic acid as a white solid (98 mg, 47%).


AnalpH2_MeOH—4 min: Rt 1.47 min; m/z 254 [M+1]+.


The following aryl boronic acid or boronic ester derivatives 43 are prepared using analogous procedures.









TABLE 24







Aryl boronic acid or boronic ester derivatives of Formula 43











Mass, % Yield,


Compound
Analytical Data
State







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AnalpH2_MeOH_4 min: Rt 0.32, 0.43 min; m/z 267 [M + 1]+
240 mg, 55%, white solid







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AnalpH2_MeOH_ 4 min: Rt 0.33, 0.61 min; m/z 307 [M + 1]+
245 mg, 49%, white solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.83 min; m/z 282 [M + 1]+
554 mg, 65%, off-white solid







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AnalpH2_MeOH_ 4 min(3): Rt 3.45 min; m/z 460 [M + 1]+
1.56 g, 96%, cream solid







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AnalpH2_MeOH_ 4 min(3): Rt 0.31 min; m/z 249 [M + 1]+
1.54 g, 54%, colourless oil







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AnalpH2_MeOH_ 4 min(3): Rt 2.76 min; m/z 516 [M + 1]+
978 mg, 64%, white solid







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AnalpH9_MeOH_ 4 min(2): Rt 1.62 min; m/z 263 [M + 1]+
242 mg, 83%, dark orange oil







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AnalpH2_MeOH_ 4 min(3): Rt 0.77 min; m/z 291 [M + 1]+
127 mg, 65%, white solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.71 min; m/z 359.5 [M + 1]+
992 mg, 46%, pale yellow solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.70 min; m/z 345 [M + 1]+
294 mg, 24%, off-white solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.72 min; m/z 356 [M + 1]+
532 mg, 74%, yellow oil







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AnalpH2_MeOH_ 4 min(3): Rt 1.66 min; m/z 331 [M + 1]+
118 mg, 25%, white solid







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AnalpH2_MeOH_ 4 min(3): Rt 3.21 min; m/z 431 [M + 1]+
899 mg, 67%, cream solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.68 min; m/z 331 [M + 1]+
233 mg, 49%, yellow solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.73 min; m/z 359 [M + 1]+
149 mg, 19%, yellow oil







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AnalpH2_MeOH_ 4 min(3): Rt 1.74 min; m/z 387 [M + 1]+
995 mg, 91%, yellow solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.77 min; m/z 373 [M + 1]+
589 mg, 81%, yellow solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.80 min; m/z 373 [M + 1]+
593 mg, 82%, dark yellow solid







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AnalpH2_MeOH_ 4 min(3): Rt 3.25 min; m/z 445 [M + 1]+
889 mg, quant., white solid







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AnalpH2_MeOH_ 4 min(3): Rt 1.75 min; m/z 359 [M + 1]+
784 mg, 73%, yellow/orange foam









Scheme JJ, Step AC (Protocol 2): Synthesis of Aryl Boronic Acid Derivatives of Formula 43 (Via Reductive Amination)
Pyridin-2-ylmethyl-[1-(4-cyclopropylmethyl-piperazine)]-5-boronic acid



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To a stirred solution of 2-formylpyridine-5-boronic acid pinacolate (200 mg, 1.33 mmol) in DCE (10 mL) was added 1-(cyclopropylmethyl)piperazine (0.217 mL, 1.46 mmol) and stirred at RT for 30 min. Sodium triacetoxyborohydride (424 mg, 2.00 mmol) was added and the reaction mixture stirred for 18 h at RT. The reaction mixture concentrated in vacuo and the residue was diluted with water (20 mL) and the aqueous layer washed with EtOAc. The combined aqueous layer was concentrated in vacuo and the crude material was purified by reverse phase preparative HPLC-MS to obtain pyridin-2-ylmethyl-[1-(4-cyclopropylmethyl-piperazine)]-5-boronic acid as a pale yellow oil (140 mg, 38%). AnalpH2_MeOH—4 min: Rt 0.33 min; m/z 275 [M+1]+.


The following aryl boronic acid derivatives 43 are prepared using analogous procedures.









TABLE 25







Aryl boronic acid derivatives of Formula 43











Mass, % Yield,


Compound
Analytical Data
State







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AnalpH9_MeOH_ 4 min: Rt 1.85 min; m/z 290 [M + 1]+
122 mg, 32%, brown oil







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AnalpH2_MeOH_ 4 min: Rt 1.72 min; m/z 362 [M + 1]+
68 mg, 47%, pale yellow solid







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AnalpH2_MeOH_ 4 min: Rt 0.73 min; m/z 253 [M + 1]+
164 mg, 55%, off-white solid









Scheme JJ, Step AC (Protocol 3): Synthesis of Aryl Boronic Ester Derivatives of Formula 43 (Via Alkylation)
2-Methyl-1-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-1H-benzoimidazole



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To a solution of 4-bromomethylphenylboronic acid pinacol ester (564 mg, 1.90 mmol) in acetone (19 mL) was added 2-methylbenzimidazole (377 mg, 2.85 mmol), potassium iodide (16 mg, 0.095 mmol) and K2CO3 (394 mg, 2.85 mmol) and the reaction mixture heated at 60° C. for 3.25 h. The reaction mixture was diluted with H2O and extracted with EtOAc (×2). The organic layers were combined, dried (phase separation cartridge) and concentrated in vacuo. The crude material was purified by reverse phase preparative HPLC-MS to afford 2-Methyl-1-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]1H-benzoimidazole as an off-white solid (234 mg, 35%).


AnalpH2_MeOH—4 min(3): Rt 2.26 min; m/z 349 [M+1]+.


The following aryl boronic ester derivatives 43 are prepared using analogous procedures.









TABLE 26







Aryl boronic ester derivatives of Formula 43











Mass,




%



Analytical
Yield,


Compound
Data
State







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AnalpH2_ MeOH_ 4 min(3): Rt 2.49 min; m/z 335 [M + 1]+.
385 mg, 53%, white solid







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Commercially available
N/A









Scheme JJ, Step AC (Protocol 3a): Synthesis of Aryl Boronic Ester Derivatives of Formula 43 (Via Alkylation)
1-[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-1H-indole



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To NaH (78 mg, 1.94 mmol) in anhydrous DMF (4 mL) under N2, at 0° C. was added indole (227 mg, 1.94 mmol) in anhydrous DMF (5 mL). The reaxtion mixture was maintained at this temperature for 10 min. 4-bromomethylphenylboronic acid pinacol ester (523 mg, 1.76 mmol) in anhydrous DMF (8 mL) was added and the reaction stirred at RT for 18 h. The reaction mixture was diluted with H2O and extracted with CH2Cl2 (×2). The organic phases were combined, washed with brine, dried (phase separation cartridge) and the solvent removed in vacuo. The crude material was purified by silica gel column chromatography eluting with isohexane and increasing the polarity to 5% EtOAc/isohexane. The compound was further purified by reverse phase preparative HPLC-MS to afford 1-[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-1H-indole as an off-white solid (120 mg, 10%).


AnalpH2_MeOH—4 min(3): Rt 3.50 min; m/z 334 [M+1]+.


The following aryl boronic ester derivatives 43 are prepared using analogous procedures.









TABLE 27







Aryl boronic ester derivatives of Formula 43











Mass, % Yield,


Compound
Analytical Data
State







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AnalpH2_MeOH_ 4 min(3): Rt 3.34 min; m/z 284 [M + 1]+
130 mg, 23%, white solid









Scheme JJ, Step AC (Protocol 3b): Synthesis of Aryl Boronic Ester Derivatives of Formula 43 (Via Alkylation)
Methyl-{1-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-azetidin-3-yl}-carbamic acid tert-butyl ester



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To 4-bromomethylboronic acid pinacol ester (500 mg, 168 mmol) and azetidin-3-yl-methyl-carbamic acid tert-butyl ester hydrochloride (561 mg, 2.52 mmol) in anhydrous THF (12 mL) was added NEt3 (704 μl, 5.05 mmol). The reaction mixture was stirred at RT, under N2 balloon, for 18 h. The reaction mixture was concentrated in vacuo, suspended in CH2Cl2 and washed with H2O. The aqueous layer was separated and washed with CH2Cl2. The organic layers were combined, dried (phase separation cartridge) and the solvent removed in vacuo. The crude material was purified by silica gel column chromatography eluting with CH2Cl2 and increasing the polarity to 20% MeOH/CH2Cl2 to afford methyl-{1-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzyl]-azetidin-3-yl}-carbamic acid tert-butyl ester as a colourless oil (441 mg, 65%).


AnalpH2_MeOH—4 min(3): Rt 2.21 min; m/z 403 [M+1]+.


Scheme JJ, Step AD (Protocol 1): Synthesis of Aryl Bromide Derivatives of Formula 47 (Via Amide Coupling)
(5-Bromo-pyrimidin-2-yl)-(4-cyclopropylmethyl-piperazin-1-yl)-methanone



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To 5-bromopyrimidine-2-carboxylic acid (100 mg, 0.49 mmol) and TBTU (158 mg, 0.49 mmol) in anhydrous DMF 94.4. mL) was added DIPEA (0.36M in CH2Cl2, 1.4 mL, 0.49 mmol) and the reaction mixture stirred at RT for 40 min. N-cyclopropylmethylpiperazine (83 mg, 0.59 mmol) in anhydrous DMF (1 mL) was added and the reaction stirred at RT for 18 h. The reaction mixture was passed through a Si—NH2 cartridge (5 g), eluting with DMF and MeOH. The eluents were combined, concentrated in vacuo and purified by reverse phase preparative HPLC-MS to obtain (5-bromo-pyrimidin-2-yl)-(4-cyclopropylmethyl-piperazin-1-yl)-methanone as a pale yellow solid (46 mg, 29%).


AnalpH2_MeOH—4 min(3): Rt 0.35, 0.81 min; m/z 325 [M+1]+.


The following bromo aryl derivatives 47 are prepared using analogous procedures.









TABLE 28







Aryl bromide derivatives of Formula 47











Mass, % Yield,


Compound
Analytical Data
State







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AnalpH2_MeOH_ 4 min(3): Rt 0.84 min; m/z 325 [M + 1]+
390 mg, 53%, colorless oil, solidifies on standing







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AnalpH9_MeOH_ 4 min(2): Rt 1.64 min; m/z 285 [M + 1]+
314 mg, 50%, tan solid







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AnalpH9_MeOH_ 4 min(2): Rt 2.06 min; m/z 284 [M + 1]+
1.17 g, 83%, dark orange oil







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AnalpH2_MeOH_ 4 min(3): Rt 3.65 min; m/z 530 [M + 1]+
971 mg, 82%, orange oil







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AnalpH2_MeOH_ 4 min(3): Rt 2.64 min; m/z 298 [M + 1]+
500 mg, 81%, white solid







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AnalpH2_MeOH_ 4 min(3): Rt 2.47 min; m/z 284 [M + 1]+
457 mg, 67%, pale yellow solid









Scheme JJ, Step AD (Protocol 1a): Synthesis of Aryl Bromide Derivatives of Formula 47 (Via Amide Coupling-via Acid Chloride)
2-(4-Bromo-phenyl)-1-[3-(tert-butyl-diphenyl-silanyloxy)-azetidin-1-yl]-ethanone



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To 4-bromophenyl acetylchloride (300 mg, 1.28 mmol) in CH2Cl2 (5 mL) was added 3-(tert-Butyl-diphenyl-silanyloxy)-azetidine (399 mg, 1.28 mmol), DIPEA (670 μL, 3.85 mmol) and the reaction stirred at RT for 2 h. The crude material was purified by silica gel column chromatography, eluting with isohexane and increasing the polarity to 80% EtOAc/isohexane to obtain 2-(4-bromo-phenyl)-1-[3-(tert-butyl-diphenyl-silanyloxy)-azetidin-1-yl]-ethanone as a colourless glass (632 mg, 97%).


AnalpH2_MeOH—4 min(3): Rt 3.71 min; m/z 510 [M+1]+.


Scheme JJ, Step AD (Protocol 3): Synthesis of Aryl Bromide Derivatives of Formula 47 (Via Alkylation)
1-(4-Bromo-benzyl)-3-(tert-butyl-diphenyl-silanyloxy)-azetidine



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To 4-bromomethylbenzyl bromide (300 mg, 1.2 mmol) in THF (5 mL) was added NEt3 (418 μl, 3 mmol) and the reaction mixture stirred at RT for 10 min. 3-(tert-Butyl-diphenyl-silanyloxy)-azetidine hydrochloride (628 mg, 1.8 mmol) was added and the reaction stirred at RT for 18 h. The reaction mixture was concentrated in vacuo and the residue partitioned between CH2Cl2 and 5% NaHCO3 (aq). The organic phase was separated, dried (MgSO4) and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with isohexane and increasing the polarity to 50% EtOAc/isohexane to obtain 1-(4-Bromo-benzyl)-3-(tert-butyl-diphenyl-silanyloxy)-azetidine as a colourless oil (328 mg, 57%).


AnalpH2_MeOH—4 min(3): Rt 2.77 min; m/z 480 [M+1]+.


Scheme JJ, Step AD (Protocol 3a): Synthesis of Aryl Bromide Derivatives of Formula 47 (Via Alkylation)
1-[1-(4-Bromo-phenyl)-1-methyl-ethyl]-azetidin-3-ol



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1-(4-Bromophenyl)-1-methyl-ethylamine (1 g, 4.67 mmol) and epichlorohydrin (439 μl, 5.6 mmol) in EtOH (15 mL) were heated at 70° C. for 18 h. The reaction mixture was concentrated in vacuo and purified by reverse phase preparative HPLC-MS to obtain 1-[1-(4-bromo-phenyl)-1-methyl-ethyl]-azetidin-3-ol as a white solid (489 mg, 38%).


AnalpH2_MeOH—4 min(3): Rt 2.77 min; m/z 480 [M+1]+.


Scheme JJ, Step AE: Synthesis of Aryl Boronic Acid or Boronic Ester Derivatives of Formula 43
2-(4-Cyclopropylmethyl-piperazine-1-carbonyl)-pyrimidine-5-boronic acid



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(5-Bromo-pyrimidin-2-yl)-(4-cyclopropylmethyl-piperazin-1-yl)-methanone (46 mg, 0.14 mmol), bis(pinacolato)diboron (43 mg, 0.17 mmol), Pd(dppf)Cl2 (12 mg, 0.014 mmol) and potassium acetate (42 mg, 0.42 mmol) in 1,4-dioxane (0.7 mL) were added to a microwave vial and the reaction mixture purged with N2 for 10 min. The reaction mixture was irradiated using a microwave reactor (300 W, 120° C., 20 min). The reaction mixture was passed through a Si-thiol cartridge (2 g) and the column washed with MeOH (4× column volumes). The solvent was removed in vacuo and purified by reverse phase preparative HPLC-MS. The sample was passed through a SCX-2 cartridge (500 mg) and the column washed with MeOH (4× column volumes). The compound was eluted from the column with 0.5M NH3/MeOH to afford 2-(4-cyclopropylmethyl-piperazine-1-carbonyl)-pyrimidine-5-boronic acid as a white solid (29 mg, 70%).


AnalpH9_MeOH—4 min(2): Rt 1.15 min; m/z 291 [M+1]+.


The following aryl boronic acid or boronic ester derivatives 43 are prepared using analogous procedures.









TABLE 29







Aryl boronic acid or boronic ester derivatives of Formula 43











Mass, % Yield,


Compound
Analytical Data
State







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AnalpH2_MeOH_ 4 min(3): Rt 0.3 min; m/z 251 [M + 1]+
163 mg, 68%, white solid







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AnalpH2_MeOH_ 4 min(3): Rt 3.72 min; m/z 556 [M + 1]+
141 mg, 47%, brown oil









Scheme J, Step AA: Synthesis of 2H-isoquinolin-1-one derivatives of formula 4 (Via Suzuki cross-coupling)
5-Methyl-3-[2-(4-methyl-piperazin-1-yl)-pyrimidin-5-yl]-2H-isoquinolin-1-one (IQ-025)



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3-Chloro-5-methyl-2H-isoquinolin-1-one (50 mg, 0.26 mmol), 2-(4-methylpiperazin-1-yl)pyrimidine-5-boronic acid pinacol ester (118 mg, 0.39 mmol), K2CO3 (73 mg, 0.52 mmol) and Pd(dppf)Cl2 (10 mg, 0.013 mmol) in DME/EtOH/H2O 4:0.5:1 (2.75 mL) were added to a microwave vial and the reaction mixture purged with N2 for 10 min. The reaction mixture was irradiated using a microwave reactor (300 W, 100° C., 60 min). The reaction mixture was filtered through celite and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with CH2Cl2 and increasing the polarity to 50% MeOH/CH2Cl2. The crude material was trituared with MeOH and washed with isohexane to afford 5-methyl-3-[2-(4-methyl-piperazin-1-yl)-pyrimidin-5-yl]-2H-isoquinolin-1-one as an off-white solid (28 mg, 32%).


AnalpH2_MeOH_QC(1): Rt 4.97 min; m/z 336 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 30







2H-isoquinolin-1-one derivatives of Formula 4













Mass,





% Yield,


Compound
Code
Analytical Data
State







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IQ-099
AnalpH2_MeOH_ QC(1): Rt 4.74 min; m/z 391 [M + 1]+
13 mg, 10%, cream solid







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IQ-071
AnalpH2_MeOH_ QC(1): Rt 6.35 min; m/z 435 [M + 1]+
31 mg, 27%, cream solid







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IQ-057
AnalpH2_MeOH_ QC(1): Rt 4.84 min; m/z 349 [M + 1]+
17 mg, 20%, pale orange solid







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IQ-076
AnalpH2_MeOH_ QC(1): Rt 6.56 min; m/z 452 [M + 1]+
72 mg, 66%, pale orange solid







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IQ-077
AnalpH2_MeOH_ QC(1): Rt 6.69 min; m/z 456 [M + 1]+
46 mg, 29%, pale brown solid







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IQ-154
AnalpH2_MeOH_ QC(1):: Rt 6.69 min; m/z 456 [M + 1]+
46 mg, 29%, pale brown solid







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IQ-080
AnalpH2_MeOH_ QC(1): Rt 5.57 min; m/z 366 [M + 1]+
39 mg, 45%, brown solid







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IQ-138
AnalpH2_MeOH_ QC(1): Rt 5.63 min; m/z 370 [M + 1]+
25 mg, 29%, white solid







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IQ-161
AnalpH2_MeOH_ QC: Rt 5.19 min; m/z 389 [M + 1]+
44 mg, 40%, pale brown solid







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IQ-162
AnalpH2_MeOH_ QC: Rt 5.41 min; m/z 404 [M + 1]+
48 mg, 43%, pale brown solid







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IQ-163
AnalpH2_MeOH_ QC: Rt 7.44 min; m/z 367 [M + 1]+
19 mg, 11%, off-white solid







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IQ-164
AnalpH2_MeOH_ QC: Rt 5.19 min; m/z 380 [M + 1]+
40 mg, 23%, beige solid







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IQ-165
AnalpH2_MeOH_ QC: Rt 5.38 min; m/z 420 [M + 1]+
50 mg, 26%, light brown solid







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AnalpH2_MeOH_ 4 min(1): Rt 3.31 min; m/z 422 [M + 1]+
Used in next step as crude material







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AnalpH2_MeOH_ 4 min: Rt 2.31 min; m/z 476 [M + 1]+
Used in next step as crude material







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IQ-166
AnalpH2_MeOH_ QC: Rt 7.23 min; m/z 366 [M + 1]+
23 mg, 20%, white solid







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IQ-229
AnalpH2_MeOH_ QC(1): Rt 4.36 min; m/z 426 [M + 1]+
4.8 mg, 15%, off-white solid







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IQ-187
AnalpH2_MeOH_ QC(1): Rt 8.05 min; m/z 395 [M + 1]+
79 mg, 42%, off-white solid







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IQ-188
AnalpH2_MeOH_ QC(1): Rt 7.10 min; m/z 335 [M + 1]+
92 mg, 27%, white solid







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IQ-225
AnalpH2_MeOH_ QC(1): Rt 4.27 min; m/z 382 [M + 1]+
7 mg, 8%, pale yellow solid







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IQ-226
AnalpH2_MeOH_ QC(1): Rt 4.74 min; m/z 384 [M + 1]+
3.6 mg, 4%, beige solid







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IQ-227
AnalpH2_MeOH_ QC(1): Rt 5.19 min; m/z 384 [M + 1]+
3.4 mg, 3%, white solid







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IQ-189
AnalpH2_MeOH_ QC(1): Rt 4.89 min; m/z 380 [M + 1]+
4.7 mg, 5%, white solid







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IQ-190
AnalpH2_MeOH_ QC(1): Rt 5.25 min; m/z 380 [M + 1]+
5 mg, 5%, white solid







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Intermediate for IQ-228
AnalpH2_MeOH_ 4 min(3): Rt 3.22 min; m/z 652 [M + 1]+
83 mg, 54%, beige solid







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Intermediate for IQ-192
AnalpH2_MeOH_ 4 min(3): Rt 3.19 min; m/z 649 [M + 1]+
63 mg, 36%, beige solid







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Intermediate for IQ-193
AnalpH2_MeOH_ 4 min(3): Rt 3.10 min; m/z 649 [M + 1]+
79 mg, 44%, beige solid







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IQ-214
AnalpH2_MeOH_ QC(1): Rt 7.24 min; m/z 349 [M + 1]+
20 mg, 17%, off-white solid







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IQ-215
AnalpH2_MeOH_ QC(1): Rt 7.45 min; m/z 367 [M + 1]+
44 mg, 37%, off-white solid







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IQ-195
AnalpH2_MeOH_ QC(1): Rt 5.35 min; m/z 408 [M + 1]+
72 mg, 38% white solid 1H NMR (400 MHz, DMSO- d6): δ11.77 (br s, 1H), 7.89 (d, J = 8.8 Hz, 2H), 7.74 (dd, J = 9.6, 2.8 Hz, 1H), 7.52, (dd, J = 9.6, 2.8 Hz, 1H), 7.48 (d, J = 7.6 Hz, 2H), 6.94 (s, 1H), 4.33-4.24 (br





s, 0.5H), 2.92-





2.73 (m, 5H),





2.61 (s, 3H),





2.22-1.93 (m,





4H), 1.92-1.55





(m, 5.5H).







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IQ-196
AnalpH2_MeOH_ QC(1): Rt 5.17 min; m/z 377 [M + 1]+
99 mg, 81%, brown solid







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  Enantiomer 1
IQ-205-1
AnalpH2_MeOH_ QC(2): Rt 4.69 min; m/z 376.5 [M + 1]+
13.4 mg, 37.5%, off-white solid; obtained via Chiral _ Method_3







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  Enantiomer 2
IQ-205-2
AnalpH2_MeOH_ QC(2): Rt 4.67 min; m/z 376.5 [M + 1]+
12.4 mg, 34.7 %, off-white solid; obtained via Chiral _ Method_3







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IQ-197
AnalpH2_MeOH_ QC(1): Rt 5.35 min; m/z 394 [M + 1]+
60 mg, 48%, off-white solid







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  Enantiomer 1
IQ-207-1
AnalpH2_MeOH_ QC(2): Rt 4.84 min; m/z 394 [M + 1]+
5.5 mg, 37%, white solid; obtained via Chiral_ Method_3







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  Enantiomer 2
IQ-207-2
AnalpH2_MeOH_ QC(2): Rt 4.83 min; m/z 394.5 [M + 1]+
4.9 mg, 33%, white solid; obtained via Chiral_ Method_3







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IQ-198
AnalpH2_MeOH_ QC(1): Rt 5.22 min; m/z 388 [M + 1]+
80 mg, 40%, off-white solid







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IQ-199
AnalpH2_MeOH_ QC(1): Rt 5.11 min; m/z 362 [M + 1]+
70 mg, 72%, off-white solid







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Intermediate for IQ-200
AnalpH2_MeOH_ 4 min(3): Rt 3.08 min; m/z 462.5 [M + 1]+
Used in next step as crude material







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Intermediate for IQ-186
AnalpH2_MeOH_ 4 min(3): Rt 3.12 min; m/z 480.5 [M +1]+
Used in next step as crude material







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IQ-201
AnalpH2_MeOH_ QC(1): Rt 5.31 min; m/z 380.4 [M + 1]+
37 mg, 34%, off-white solid







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IQ-202
AnalpH2_MeOH_ QC(1): Rt 5.11 min; m/z 362 [M + 1]+
16 mg, 16%, white solid







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IQ-203
AnalpH2_MeOH_ QC(1): Rt 5.35 min; m/z 381 [M + 1]+
16 mg, 15%, white solid







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IQ-204
AnalpH2_MeOH_ QC(1): Rt 5.29 min; m/z 390.5 [M + 1]+
57 mg, 42%, off-white solid







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IQ-175
AnalpH2_MeOH_ QC(1): Rt 5.30 min; m/z 376.5 [M + 1]+
131 mg, 49%, off-white solid







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IQ-176
AnalpH2_MeOH_ QC(1): Rt 5.48 min; m/z 404.5 [M + 1]+
43 mg, 31%, off-white solid







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IQ-20
AnalpH2_MeOH_ QC(2): Rt 4.91, min; m/z 418.5 [M + 1]+
28 mg, 13% white solid







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Intermediate for IQ-177
AnalpH2_MeOH_ 4 min(3): Rt 2.16 min; m/z 434.5 [M + 1]+
154 mg, 83%, brown, sticky solid







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Intermediate for IQ-178
AnalpH2_MeOH_ 4 min(3): Rt 2.21 min; m/z 452.5 [M + 1]+
159 mg, 83%, brown, sticky solid







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IQ-208
AnalpH2_MeOH_ QC(2): Rt 4.96 min; AnalpH2_MeOH_ 4 min(2): m/z 422 [M + 1]+
42 mg, 19%, white solid







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IQ-209
AnalpH2_MeOH_ QC(2): Rt 4.89 min; AnalpH2_MeOH_ 4 min(2): m/z 404 [M + 1]+
86 mg, 38%, beige solid







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IQ-210
AnalpH2_MeOH_ QC(2): Rt 5.02 min; AnalpH2_MeOH_ 4 min(2): m/z 422.5 [M + 1]+
92 mg, 29%, off-white solid 1H NMR (400 MHz, DMSO- d6): δ11.72 (br s, 1H), 7.88 (d, J = 8.3 Hz, 2H), 7.74 (br dd, J = 9.3, 2.7 Hz, 1H), 7.55-7.44 (m, 3H), 6.93 (s, 1H), 3.63 (br d, J = 13.2 Hz, 1H), 3.44 (br d, J = 13.2 Hz, 1H), 3.00 (br s, 3H), 2.60 (s, 3H), 2.48-





2.44 (m, 1H),





2.29 (d, J =





9.1 Hz, 1H),





2.24 (s, 3H),





2.13 (br s,





1H), 1.86-1.79





(m, 1H), 1.56-





1.50 (m, 1H),





1.13 (s, 3H),





0.89 (br s,





1H).







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IQ-211
AnalpH2_MeOH_ QC(2): Rt 4.76 min; m/z 390.5 [M + 1]+
22 mg, 10%, off-white solid







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IQ-212
AnalpH2_MeOH_ QC(2): Rt 4.89 min; AnalpH2_MeOH_ 4 min(2): m/z 408 [M + 1]+
84 mg, 37%, off-white solid







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Intermediate for IQ-213
AnalpH2_MeOH_ 4 min(3): Rt 3.16 min; m/z 494 [M + 1]+
Used in next step as crude material







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IQ-180
AnalpH2_MeOH_ QC(2): Rt 4.75 min; m/z 316 [M + 1]+
140 mg, 79%, beige solid







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IQ-179
AnalpH2_MeOH_ QC(2): Rt 5.81 min; m/z 380.5 [M + 1]+
66 mg, 31%, beige solid







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IQ-181
AnalpH2_MeOH_ QC(2): Rt 6.56 min; m/z 366.5 [M + 1]+
101 mg, 49%, beige solid







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IQ-183
AnalpH2_MeOH_ QC(2): Rt 8.54 min; m/z 365.5 [M + 1]+
17 mg, 16%, beige solid







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IQ-184
AnalpH2_MeOH_ QC(2): Rt 8.16 min; m/z 315 [M + 1]+
10.6 mg, 5%, brown solid









Scheme J, Step AF (Protocol 1): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 5 (Via BOC deprotection)
7-Fluoro-5-methyl-3-(4-piperazin-1-ylmethyl-phenyl)-2H-isoquinolin-1-one (IQ-078)



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The synthesis is analogous to the Boc deprotection procedure used in Scheme A, Step C (Protocol 1) above to give 7-Fluoro-5-methyl-3-(4-piperazin-1-ylmethyl-phenyl)-2H-isoquinolin-1-one as an off-white solid (18.4 mg, 37%).



1H NMR (400 MHz, DMSO-d6): δ7.86 (d, J=8 Hz, 2H), 7.81 (dd, J=9, 3 Hz, 1H), 7.60 (dd, J=9, 3 Hz, 1H), 7.50 (d, J=8 Hz, 2H), 6.94 (s, 1H), 3.57 (s, 2H), 2.79-2.77 (m, 4H), 2.68 (s, 3H), 2.39 (br s, 4H).


AnalpH2_MeOH_QC(1): Rt 5.49 min; m/z 352 [M+1]+


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 31







2H-isoquinolin-1-one Formula 5












Analytical
Mass, % Yield,


Compound
Reference
Data
State







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IQ-079
AnalpH2_ MeOH_QC(1): Rt 5.05 min; m/z 356 [M + 1]+
8 mg, 14%, off-white solid







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IQ-158
AnalpH2_ MeOH_ QC: Rt 5.62 min; m/z 356 [M + 1]+
22 mg, 65% white solid







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IQ-072
AnalpH2_ MeOH_ QC(1): Rt 4.87 min; m/z 335 [M + 1]+
11 mg, 50%, white solid







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IQ-026
AnalpH2_ MeOH_ QC(1): Rt 5.05 min; m/z 322 [M + 1]+
28 mg, 17%, white solid







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IQ-160
AnalpH2_ MeOH_ QC: Rt 4.31 mins; m/z 375 [M + 1]+
16 mg, 29% light brown solid







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IQ-200
AnalpH2_ MeOH_ QC(1): Rt 5.21 min; m/z 362.5 [M + 1]+
210 mg, 76%, off-white solid







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IQ-186
AnalpH2_ MeOH_ QC(1): Rt 5.39 min; m/z 380.5 [M + 1]+
135 mg, 61%, off-white solid







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IQ-177
AnalpH2_ MeOH_ QC(2): Rt 3.68 min; m/z 334.5 [M + 1]+
35 mg, 29%, white solid







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IQ-178
AnalpH2_ MeOH_ QC(2): Rt 3.84 min; m/z 352.5 [M + 1]+
42 mg, 34%, white solid







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IQ-213
AnalpH2_ MeOH_ QC(2): Rt 4.81 min; AnalpH2_ MeOH_ 4min(2): m/z 394 [M + 1]+
80 mg, 36%, off-white solid









Scheme J, Step AF (Protocol 3): Synthesis of 2H-isoquinolin-1-one Derivatives of formula 5 (Via TBDPS deprotection)
5,7-Difluoro-3-{4-[4-(2-hydroxy-ethyl)-piperazine-1-carbonyl]-phenyl}-2H-isoquinolin-1-one (IQ-228)



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The synthesis is analogous to the TBDPS deprotection procedure used in Scheme A, Step C (Protocol 3) above to give 5,7-difluoro-3-{4-[4-(2-hydroxy-ethyl)-piperazine-1-carbonyl]-phenyl}-2H-isoquinolin-1-one as a white solid (20 mg, 48%).



1H NMR (400 MHz, DMSO-d6): δ11.96 (br s, 1H), 7.87 (d, J=8 Hz, 2H), 7.80-7.75 (m, 2H), 7.50 (d, J=8 Hz, 2H), 6.91 (s, 1H), 4.46-4.44 (m, 1H), 3.63 (br s, 2H), 3.50 (q, J=7 Hz, 2H), 3.35 (br s, 6H), 2.42 (t, J=6 Hz, 2H).


AnalpH2_MeOH_QC(1): Rt 5.19 min; m/z 414 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 32







2H-isoquinolin-1-one Formula 5












Analytical
Mass, % Yield,


Compound
Reference
Data
State







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IQ-192
AnalpH2_ MeOH_ QC(3): Rt 9.23 min; m/z 410 [M + 1]+
26 mg, 64%, white solid







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IQ-193
AnalpH2_ MeOH_ QC(1): Rt 4.90 min; m/z 410 [M + 1]+
37 mg, 75%, white solid









N,N-Bis-(2-hydroxy-ethyl)-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzamide (IQ-191)



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To a stirred solution of N,N-Bis-(2-methoxy-ethyl)-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzamide (40 mg, 0.10 mmol) in CH2Cl2 (2.5 mL) under N2 at −78° C. was added boron tribromide (1M in CH2Cl2, 2.54 mL, 2.54 mmol). The reaction was allowed to warm to RT and stirred for 16 h. The reaction mixture was quenched with H2O and extracted with EtOAc (5 mL) upon which a pale yellow solid precipitated and was filtered off. The aqueous phase was further extracted with CH2Cl2 (5 mL). The combined organics were evaporated to dryness. Product was found to be also present in the aqueous phase and was passed through an Isolute-103 cartridge (500 mg), washing with H2O (4× column volumes). The product was eluted from the column with MeOH (4× column volumes) and evaporated to dryness. The combined crude product was purified by reverse phase preparative HPLC-MS to obtain N,N-bis-(2-hydroxy-ethyl)-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzamide as a white solid (23 mg, 63%).



1H NMR (400 MHz, DMSO-d6): δ11.64 (br s, 1H), 8.09 (d, J=8 Hz, 1H), 7.88 (d, J=8 Hz, 2H), 7.58 (d, J=7 Hz, 1H), 7.52 (d, J=8.6 Hz, 2H), 7.39 (t, J=7.6 Hz, 1H), 6.92 (s, 1H), 4.88-4.82 (m, 2H), 3.66-3.62 (m, 2H), 3.56-3.53 (m, 2H), 3.49-3.48 (m, 2H), 2.58 (s, 3H).


AnalpH2_MeOH_QC(1): Rt 6.76 min; m/z 367 [M+1]+.


General Procedure for Synthesis of 2H-Isoquinolin-1-Ones Amide Derivatives of Formula 4



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Scheme K, Step AG: Synthesis of 4-(5-Methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoic acid



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3-Chloro-5-methyl-2H-isoquinolin-1-one (50 mg, 0.26 mmol), 4-carboxybenezeneboronic acid (64 mg, 0.39 mmol), K2CO3 (73 mg, 0.52 mmol) and Pd(dppf)Cl2 (11 mg, 0.013 mmol) in DME/EtOH/H2O 4:0.5:1 (2.75 mL) were added to a microwave vial and the reaction mixture purged with N2 for 10 min. The reaction mixture was irradiated using a microwave (300 W, 120° C., 2 h). The reaction mixture was concentrated in vacuo, water added and the mixture acidified to pH2 with 0.2M HCl aq. A brown solid precipiated from the solution which was filtered and dried in vacuo, dissolved in DMF and passed through a Si-thiol cartridge, eluting with DMF (4× column volumes) and the solvent removed in vacuo. The resulting solid was triturated with MeOH, filtered and dried to give 4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-benzoic acid as a beige solid (29 mg, 40%).


AnalpH2_MeOH_QC(1): Rt 7.93 min; m/z 280 [M+1]+.


Scheme K, Step AE: Synthesis of N-Methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(1-methyl-piperidin-4-ylmethyl)-benzamide (IQ-095)



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The synthesis is analogous to the acid coupling procedure used in Step E above to give N-methyl-4-(5-methyl-1-oxo-1,2-dihydro-isoquinolin-3-yl)-N-(1-methyl-piperidin-4-ylmethyl)-benzamide as a pale yellow foam (27 mg, 93%).



1H NMR (400 MHz, DMSO-d6): δ11.65-11.59 (br s, 1H), 8.09 (d, J=8 Hz, 1H), 7.89 (d, J=8 Hz, 2H), 7.57 (d, J=7 Hz, 1H) 7.51 (br d, J=7 Hz, 1H), 7.45 (br d, J=7 Hz, 1H) 7.39 (t, J=7 Hz, 1H), 6.92 (s, 1H), 3.39-3.37 (m, 1H), 3.18-3.14 (m, 1H), 2.98 (s, 1H), 2.93 (s, 2H), 2.78 (d, J=10 Hz, 1H), 2.66 (d, J=10 Hz, 1H), 2.58 (s, 3H), 2.16 (s, 2H), 2.08 (s, 1H), 1.88-1.63 (m, 4H), 1.48-1.44 (d, J=10 Hz, 1H), 1.28-1.21 (m, 1H), 0.89-0.80 (m, 1H).


AnalpH2_MeOH_QC(1): Rt 5.18 min; m/z 404 [M+1]+.


General Procedure for Synthesis of 2H-Isoquinolin-1-Ones Amide Derivatives of Formula 4 & 5 (Via Route 2a)



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Scheme L, Step AI: Synthesis of Aryl Boronic Acid Derivatives of Formula 48
5-Methyl-1-oxo-1,2-dihydro-isoquinoline-3-boronic acid



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3-Chloro-5-methyl-2H-isoquinolin-1-one (100 mg, 0.52 mmol), bis(pinacolato)diboron (157 mg, 0.62 mmol), Pd(dppf)Cl2 (42 mg, 0.054 mmol) and KOAc (153 mg, 1.56 mmol) in 1,4-dioxane (2 mL) were added to a microwave vial and the reaction mixture purged with N2 for 10 min. The reaction mixture was irradiated using a microwave reactor (300 W, 120° C., 20 min). The reaction mixture was passed through a Si-thiol cartridge and concentrated in vacuo. The crude product was purified by reverse phase preparative HPLC-MS to afford 5-methyl-1-oxo-1,2-dihydro-isoquinoline-3-boronic acid as a white solid (51 mg, 49%).


AnalpH2_MeOH—4 min(3): Rt 2.18 min; m/z 204 [M+1]+.


The following boronic acid derivatives 48 are prepared using analogous procedures.









TABLE 33







Boronic acid derivatives of Formula 48











Mass,




% Yield,


Compound
Analytical Data
State







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AnalpH2_MeOH_ 4 min(3): Rt 2.18 min; m/z 222 [M + 1]+
59 mg, 38%, pale yellow solid







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AnalpH2_MeOH_ 4 min(3): Rt 2.41 min; m/z 222 [M + 1]+
84 mg, 40%, off- white solid









Scheme L, Step AJ: Synthesis of 2H-isoquinolin-1-one Derivatives of Formula 4 (Via Suzuki Cross-Coupling)
3-[5-(4-Cyclopropylmethyl-piperazine-1-carbonyl)-pyridin-2-yl]-5-methyl-2H-isoquinolin-1-one (IQ-223)



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5-Methyl-1-oxo-1,2-dihydro-isoquinoline-3-boronic acid (40 mg, 0.20 mmol), (6-bromo-pyridin-3-yl)-(4-cyclopropylmethyl-piperazin-1-yl)-methanone (95 mg, 0.30 mmol), K2CO3 (56 mg, 0.4 mmol) and Pd(dppf)Cl2 (16 mg, 0.02 mmol) in DME/EtOH/H2O 4:0.5:1 (3.5 mL) were added to a microwave vial and the reaction mixture purged with N2 for 10 min. The reaction mixture was irradiated using a microwave reactor (300 W, 130° C., 60 min). The reaction mixture was filtered through a Si-thiol and concentrated in vacuo. The crude material was purified by reverse phase preparative HPLC-MS to obtain 3-[5-(4-cyclopropylmethyl-piperazine-1-carbonyl)-pyridin-2-yl]-5-methyl-2H-isoquinolin-1-one as a brown solid (18 mg, 22%).


AnalpH2_MeOH_QC(1): Rt 4.97 min; m/z 403 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 34







2H-isoquinolin-1-one derivatives of Formula 4












Analytical
Mass, % Yield,


Compound
Code
Data
State







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IQ-224
AnalpH2_ MeOH_QC(1): Rt 4.41 min; m/z 364 [M + 1]+
3.2 mg, 16%, off-white solid







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IQ-220
AnalpH2_ MeOH_ QC(3): Rt 8.03 min; m/z 363 [M + 1]+
34 mg, 34%, white solid







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IQ-221
AnalpH2_ MeOH_ QC(3): Rt 7.66 min; m/z 381 [M + 1]+
78 mg, 76%, beige solid







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IQ-222
AnalpH2_ MeOH_QC(1): Rt 4.96 min; m/z 381 [M + 1]+
35 mg, 24%, beige solid







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IQ-194
AnalpH2_ MeOH_QC(1): Rt 7.15 min; m/z 371 [M + 1]+
32 mg, 16%, white solid







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Intermediate for IQ-170
AnalpH2_ MeOH_ 4 min(3): Rt 2.80 min; m/z 577 [M + 1]+
300 mg, quant., black oil







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IQ-217
AnalpH2_ MeOH_QC(1): Rt 7.87 min; m/z 378 [M + 1]+
41 mg, 32%, pale brown solid







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IQ-218
AnalpH2_ MeOH_QC(1): Rt 8.00 min; m/z 396 [M + 1]+
37 mg, 27%, off- white solid







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IQ-216
AnalpH2_ MeOH_QC(1): Rt 7.74 min; m/z 381 [M + 1]+
140 mg, 69%, off-white solid







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IQ-185
AnalpH2_ MeOH_QC(1): Rt 5.54 min; m/z 367 [M + 1]+
14.4 mg, 5.5%, white solid 1H NMR (400 MHz, DMSO-d6): δ11.68 (br s, 1H), 7.76 (d, J = 8.8 Hz, 2H), 7.58 (dd, J = 9.2, 2.8 Hz, 1H), 7.58 (d, J = 8.8 Hz, 2H), 7.51 (dd, J = 9.6, 2.8 Hz, 1H), 6.85





(s, 1H), 5.22 (d,





J = 6.4 Hz, 1H),





4.22-4.15 (m,





1H), 3.25 (dd, J =





7.2, 6.0 Hz, 2H),





2.87 (dd, J = 7.2,





6.0 Hz, 2H), 2.59





(s, 3H), 1.28 (s,





6H).









Scheme L, Step AK: Synthesis of 2H-Isoquinolin-1-One Derivatives of Formula 5 (Via TBDPS Deprotection)
7-Fluoro-3-[4-(3-hydroxy-azetidin-1-ylmethyl)-phenyl]-5-methyl-2H-isoquinolin-1-one (IQ-170)



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The synthesis is analogous to the TBDPS deprotection procedure used in Scheme A, Step C (Protocol 3) above to give 7-Fluoro-3-[4-(3-hydroxy-azetidin-1-ylmethyl)-phenyl]-5-methyl-2H-isoquinolin-1-one as a white solid (3 mg, 11%).


AnalpH2_MeOH_QC(1): Rt 5.25 min; m/z 339 [M+1]+.


General Procedure for Synthesis of 2H-Isoquinolin-1-One Acetylene Derivatives of Formula 51 & 52



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Scheme M, Step AL: Synthesis of 2H-Isoquinolin-1-One Derivatives of Formula 50
5-Bromo-3-[4-(2-dimethylaminoethoxy)phenyl]-2H-isoquinolin-1-one (IQ-237)



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The synthesis is analogous to the cyclisation procedure used in Scheme A Step B (protocol 3) above to give 5-bromo-3-[4-(2-dimethylaminoethoxy)phenyl]-2H-isoquinolin-1-one as a yellow solid (1.23 g, 86%).



1H NMR (400 MHz, DMSO-d6): δ11.78 (br s, 1H), 8.21 (d, J=8 Hz, 1H), 8.02 (d, J=8 Hz, 1H), 7.72 (d, J=8.8 Hz, 2H), 7.37 (t, J=7.8 Hz, 1H), 7.07 (d, J=8.8 Hz, 2H), 6.80 (s, 1H), 4.11 (t, J=5.8 Hz, 2H), 2.64 (t, J=5.8 Hz, 2H), 2.22 (s, 6H).


AnalpH2_MeOH_QC: Rt 5.69 min; m/z 387 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 35







2H-isoquinolin-1-one derivatives of Formula 4













Mass, % Yield,


Compound
Code
Analytical Data
State







embedded image


IQ-238
AnalpH2_ MeOH_4 min: Rt 1.97 min; m/z 412 [M + 1]+
845 mg, 79%, pale orange/pink solid









Scheme M, Step AM (Protocol 1): Synthesis of 2H-Isoquinolin-1-One Derivatives of Formula 51
3-[4-(2-Dimethylaminoethoxy)phenyl]-5-(4-hydroxybut-1-ynyl)-2H-isoquinolin-1-one (IQ-236)



embedded image


5-Bromo-3-[4-(2-dimethylaminoethoxy)phenyl]-2H-isoquinolin-1-one (50.0 mg, 0.130 mmol), triethylamine (1.1 mL, 8.36 mmol), dichlorobis(triphenylphosphine)palladium(II) (5.0 mg, 0.0065 mmol), copper (I) iodide (3.0 mg, 0.009 mmol), 3-butyn-1-ol (20 μL, 0.260 mmol) in DMF (1.1 mL) were added to a microwave vial and the reaction mixture purged with N2 for 5 min. The reaction mixture was irradiated using a microwave reactor (300 W, 100° C., 90 min). The reaction mixture was filtered through Celite® 545, diluted with DMSO and was purified by reverse phase preparative HPLC-MS to obtain 3-[4-(2-dimethylaminoethoxy)phenyl]-5-(4-hydroxybut-1-ynyl)-2H-isoquinolin-1-one as a brown solid (19.0 mg, 39%).



1H NMR (400 MHz, CDCl3): δ9.77 (br s, 1H), 8.31 (d, J=8.4 Hz, 1H), 7.75 (dd, J=7.6, 1.0 Hz, 1H), 7.64 (d, J=8.8 Hz, 2H), 7.37 (t, J=7.8 Hz, 1H), 7.17 (s, 1H), 7.03 (d, J=8.8 Hz, 2H), 4.29 (t, J=5.2 Hz, 2H), 3.91 (t, J=6.2 Hz, 2H), 3.12 (t, J=4.4 Hz, 2H), 2.83 (t, J=6.2 Hz, 2H), 2.60 (s, 6H).


AnalpH2_MeOH_QC: Rt 5.17 min; m/z 377 [M+1]+.


The following 2H-isoquinolin-1-one derivatives are prepared using analogous procedures.









TABLE 36







2H-isoquinolin-1-one Formula 51













Mass, % Yield,


Compound
Code
Analytical Data
State







embedded image


IQ-232
AnalpH2_ MeOH_QC: Rt 4.93 min; m/z 363 [M + 1]+
19 mg, 20%, brown oil







embedded image


IQ-234
AnalpH2_ MeOH_QC: Rt 5.53 min; m/z 377 [M + 1]+
36 mg 34% beige solid







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IQ-233
AnalpH2_ MeOH_QC(1): Rt 5.70 min; m/z 402 [M + 1]+
23 mg, 44%, beige solid







embedded image


IQ-231
AnalpH2_ MeOH_QC(1): Rt 5.06 min; m/z 388 [M + 1]+
25 mg, 41%, beige solid







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IQ-235
AnalpH2_ MeOH_4 min: Rt 0.94 min m/z 415 [M + 1]+
36 mg, 54%, off- white solid









Scheme M, Step AM (Protocol 2): Synthesis of 2H-Isoquinolin-1-One Derivatives of Formula 51
4-[4-(1-Oxo-5-trimethylsilanylethynyl-1,2-dihydroisoquinolin-3-yl)benzyl]piperazine-1-carboxylic acid tert-butyl ester



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4-[4-(5-Bromo-1-oxo-1,2-dihydro-isoquinolin-3-yl)benzyl]piperazine-1-carboxylic acid (140.0 mg, 0.281 mmol), ethynyltrimethylsilane (119 μL, 0.843 mmol), triethylamine (391 μL, 2.81 mmol), dichlorobis(triphenylphosphine)palladium(II) (19.6 mg, 0.028 mmol), and triphenylphosphine (3.67 mg, 0.014 mmol) in anhydrous DMF (3 mL) were added to a microwave vial and the reaction mixture purged with N2 for 5 min. Copper (I) iodide (5.33 mg, 0.028 mmol) was added and the mixture was degassed for a further minute. The reaction mixture was irradiated using a microwave reactor (300 W, 110° C., 1 h). The reaction mixture was then concentrated in vacuo, diluted with water (50 mL) and extracted with EtOAc (2×50 mL). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography, eluting with iso-hexane and increasing the polarity to 100% EtOAc/iso-hexane to afford 4-[4-(1-oxo-5-trimethylsilanylethynyl-1,2-dihydroisoquinolin-3-yl)benzyl]piperazine-1-carboxylic acid tert-butyl ester as a yellow solid (70.3 mg, 49%).


AnalpH2_MeOH—4 min (3): Rt 3.05 min; m/z 515.5 [M+1]+.


Scheme M, Step AN (Step 1): Synthesis of 2H-Isoquinolin-1-One Derivatives of Formula 52
4-[4-(5-Ethynyl-1-oxo-1,2-dihydroisoquinolin-3-yl)benzyl]piperazine-1-carboxylic acid tert-butyl ester



embedded image


To a stirred solution of 4-[4-(1-oxo-5-trimethylsilanylethynyl-1,2-dihydroisoquinolin-3-yl)benzyl]piperazine-1-carboxylic acid tert-butyl ester compound (70.0 mg, 0.136 mmol) in THF (5 mL) was added TBAF (1M in THF, 272 μL, 0.272 mmol). The resulting reaction mixture was stirred at RT for 2 h and then quenched by the addition of water (20 mL). The mixture was extracted with EtOAc (3×20 mL) and the combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated in vacuo to obtain 4-[4-(5-ethynyl-1-oxo-1,2-dihydroisoquinolin-3-yl)benzyl]piperazine-1-carboxylic acid tert-butyl ester (60.3 mg, 100%) as an orange solid. The crude compound was used for the next step without further purification.


AnalpH2_MeOH—4 min (3): Rt 2.35 min; m/z 444.5 [M+1]+.


Scheme M, Step AN (Step 2): Synthesis of 2H-Isoquinolin-1-One Derivatives of Formula 52
5-Ethynyl-3-(4-piperazin-1-ylmethylphenyl)-2H-isoquinolin-1-one (IQ-230)



embedded image


4-[4-(5-Ethynyl-1-oxo-1,2-dihydroisoquinolin-3-yl)benzyl]piperazine-1-carboxylic acid tert-butyl ester (60.3 mg, 0.136 mmol) and 4M HCl/dioxane (3 mL) in CH2Cl2 (3 mL) were stirred at RT for 2 h. The reaction mixture was concentrated in vacuo and the crude material was purified by reverse phase preparative HPLC-MS to obtain 5-ethynyl-3-(4-piperazin-1-ylmethylphenyl)-2H-isoquinolin-1-one as an off-white solid (16.0 mg, 34%).



1H NMR (400 MHz, DMSO-d6): δ8.23 (d, J=8 Hz, 1H), 7.89 (dd, J=7.6 Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.47 (t, J=7.6 Hz, 1H), 7.43 (d, J=8.0 Hz, 2H), 6.97 (s, 1H), 4.67 (s, 1H), 3.48 (s, 2H), 2.69-2.67 (m, 4H), 2.30 (br s, 4H).


AnalpH2_MeOH_QC (1): Rt 5.46 min; m/z 344.5 [M+1]+.


Biological Methods
Biochemical Assay 1:
TNKS1/PARP Biochemical Assay

Tankyrase activity was assayed using a 96-well format HT Universal Chemiluminescent PARP Assay Kit (Trevigen, Inc, cat. no. 4676-096-K) according to the manufacturer's instructions. In short, tankyrase/PARP activity is quantified by the incorporation of biotinylated nicotinamide adenine dinucleotide (biotin-NAD+) onto the immobilised pseudo substrate, Histone. The extent of poly(Biotin-ADP)ribosylation (PARylation) in the presence of increasing dose of inhibitor is then quantified by binding of streptavidin conjugated horse radish peroxidase (strep-HRP) followed by chemiluminescent detection.


Prior to assay initiation, inhibitor stocks were prepared in aqueous DMSO (10% (v/v)) from 5 millimolar (mM) stock in 100% DMSO (Sigma Aldrich, cat. no. 265855) as 10× concentrations. For the primary assay (i.e., single dose at 1 micromolar (μM) final concentration) this corresponded to 10 μM in 10% DMSO. For IC50 determination, this corresponded to 100 μM, 30 μM, 10 μM, 3.0 μM, 1.0 μM, 0.30 μM, 0.10 μM and 0 μM in 10% DMSO for final concentrations of 10 μM, 3.0 μM, 1.0 μM, 0.30 μM, 0.10 μM, 0.030 μM, 0.010 μM and 0 μM with 1% (v/v) final DMSO. The assay was initiated by the addition of 10× inhibitor (5 microlitres (μL)) or 10% aqueous DMSO (5 μL) to triplicate wells. Twenty microlitres of diluted TNKS1 protein (200 nanomolar (nM) final conc.) in PARP buffer (Trevigen, Inc, cat. no. 4671-096-02) was added to each histone coated well, which was previously hydrated with PARP buffer. Triplicate wells with 1% DMSO/buffer alone (no enzyme) were also added as a measure of assay ‘noise’. Positive control for PARP inhibition included the addition of 4-amino-1,8-naphthalimide (Sigma Aldrich, cat. no A0966) in corresponding doses.


The mixture was incubated for 10 minutes at room temperature and the PARylation reaction initiated by the addition of PARP cocktail (25 μL, Trevigen, Inc) containing biotin-NAD+ (Trevigen, Inc, cat. no. 4670-500-01), activated DNA (Trevigen, Inc, cat. no. 4671-096-06) and PARP buffer. The reaction was incubated for 1.5 hours (for TNKS1) or 1 hour (for PARP1) at room temperature. The reaction mixture was then removed by aspiration and the wells washed (3×200 μL) with phosphate buffered saline containing Triton X-100 (0.1% (v/v), Sigma Aldrich cat. no. T8787). The wells were then washed (3×200 μL) with phosphate buffered saline and then incubated with strep-HRP (50 μL, Trevigen, Inc, cat. no. 4800-30-06) in strep-diluent (1:500 dilution, Trevigen Inc, cat. no. 4671-096-04) for 1 hour at room temperature. The Strep-HRP mixture was then aspirated and the wells washed (3×200 μL) with phosphate buffered saline containing Triton X-100 (0.1% (v/v)) followed by phosphate buffered saline (3×200 μL) and then incubated with PeroxyGlow™ reagent (100 μL, Trevigen, Inc, cat. nos. 4675-096-01, 4675-096-02, room temperature, mixed 1:1).


The amount of light emitted as a result of the peroxidase-chemiluminescent reagent reaction was in proportion to the extent of poly(Biotin-ADP)ribosylation and was immediately measured with a Victor2 plate reader (Perkin Elmer, luminescence detection assay, luminescent units described as ‘Counts Per Second’ (CPS)). The data were normalised to the DMSO control after subtraction of ‘noise’ and was expressed as % PARP activity as a function of inhibitor dose. Inhibition was expressed as 100%−(% PARP activity). Dose response curves used to determine IC50 values were Log transformed and analysed by non-linear regression analysis (variable slope) using Prism (GraphPad Software, Inc) and were presented as IC50 with 95% confidence interval to determine relative potency.


Preparation of Recombinant Proteins:

Tankyrase1 (pNIC-Bsa-4-TNKS1PARP) expression construct was obtained from the Structural Genomics Consortium (SGC) and expresses the active PARP domain of TNKS1 as a polyhistidine tagged protein. The expression and purification of TNKS1 protein was carried out according to the SGC protocol provided at http://www.thesgc.org/structures/materials_methods/2RF5/, which is summarised in the following table.











Structure
TNKS1






PDB Code
2RF5





Entry clone
BC098394


accession






Entry clone
Mammalian Gene Collection


source






Tag
N-terminal hexahistidine tag with integrated TEV protease cleavage site:



mhhhhhhssgvdlgtenlyfq*s(m)





Construct
mhhhhhhssgvdlgtenlyfq*sMQGTNPYLTFHCVNQGTILLDLAPEDKEYQS


sequence
VEEEMQSTIREHRDGGNAGGIFNRYNVIRIQKVVNKKLRERFCHRQKE



VSEENHNHHNERMLFHGSPFINAIIHKGFDERHAYIGGMFGAGIYFAEN



SSKSNQYVYGIGGGTGCPTHKDRSCYICHRQMLFCRVTLGKSFLQFSTI



KMAHAPPGHHSVIGRPSVNGLAYAEYVIYRGEQAYPEYLITYQIMKPEA



PSQTATAAEQ





Vector
pNIC-Bsa4





Expression

E.coli Rosetta2(DE3) (Novagen)


host






Growth
Cells from a glycerol stock were streaked onto LB-agar plates. 5-10


method
colonies were used to inoculate 20 mL TB supplemented with 8 g/l



glycerol, 100 μg/mL kanamycin and 34 μg/mL chloramphenicol. The



cells were grown at 30° C. overnight. The overnight culture (20 mL) was



used to inoculate 1.5 I TB supplemented with 8 g/l glycerol, 50 μg/mL



kanamycin and approximately 200 μl PPG P2,000 81380 anti-foam



solution (Fluka). The culture was grown in a LEX bioreactor system



(Harbinger Biotechnology) at 37° C. until OD600 reached ~2. The culture



was down-tempered to 18° C. over a period of 1 hour before target



expression was induced by addition of 0.5 mM IPTG. Expression was



allowed to continue overnight and cells were harvested the following



morning by centrifugation (5,500 × g, 10 min, 4° C.). The resulting cell



pellet (38.2 g wet cell weight) was resuspended in lysis buffer 



(2 mL/g cell pellet), supplemented with one tablet of Complete EDTA-free



protease inhibitor (Roche Applied Science). The cell suspension was 



stored at −80° C.





Extraction
Lysis buffer: 50 mM HEPES, 300 mM NaCl, 10% glycerol, 10 mM


buffers
imidazole, 0.5 mM TCEP, pH 7.8





Extraction
The cell suspension was quickly thawed in water and 2500 U Benzonase


procedure
(Merck) was added. Cells were disrupted by sonication (Vibra-Cell,



Sonics) at 80% amplitude for 3 min effective time (pulsed 4s on, 4s off)



and cell debris was removed by centrifugation (49,100 × g, 20 min, 4° C.).



The supernatant was decanted and filtered through a 0.45 μm flask filter.





Purification
IMAC wash1 buffer: 30 mM HEPES, 500 mM NaCl, 10% glycerol, 10 mM


buffers
imidazole, 0.5 mM TCEP, pH 7.5.



IMAC wash2 buffer: 30 mM HEPES, 500 mM NaCl, 10% glycerol, 25 mM



imidazole, 0.5 mM TCEP, pH 7.5.



IMAC elution buffer: 30 mM HEPES, 500 mM NaCl, 10% glycerol, 500



mM imidazole, 0.5 mM TCEP, pH 7.5.



Gel filtration (GF) buffer: 30 mM HEPES, 300 mM NaCl, 10% glycerol,



0.5 mM TCEP, pH 7.5





Purification
Columns:


procedure
IMAC: Ni-charged 1 mL HiTrap Chelating HP (GE Healthcare). Gel



filtration column: HiLoad 16/60 Superdex 75 Prep Grade (GE



Healthcare).



Procedure:



Purification of the protein was performed as a two step process on an



ÄKTAxpress system (GE Healthcare). Prior to purification, columns were



equilibrated with IMAC wash1 buffer and gel filtration buffer, 



respectively. The filtered lysate was loaded onto the Ni-charged



HiTrap Chelating



column and washed with IMAC wash1 buffer followed by IMAC wash2



buffer. Bound protein was eluted from the IMAC column with IMAC



elution buffer and automatically loaded onto the gel filtration column.



Fractions containing the target protein were pooled and fresh TCEP was



added to a final concentration of 2 mM. The protein was subsequently



concentrated using a Amicon Ultra-15 centrifugal filter device, 10,000



NMWL (Millipore) to 22.8 mg/mL in a volume of 0.28 mL. The identity of



the protein was confirmed by mass spectrometry.






Tankyrase2 (pNIC-Bsa-4-TNKS2PARP) expression construct was also obtained from the Structural Genomics Consortium (SGC) and prepared in an analogous method to TNKS1.


PARP1 protein was commercially available and was obtained from Trevigen, Inc (PARP-HSA ‘High Specific Activity’, cat. no. 4668-50-010).


Cell-Based Assay 1:
Wnt-Luciferase Reporter Assay
Generation of Reporter Cell Lines:

A Wnt dependent cell line (i.e., DLD1 colorectal adenocarcinoma cell line) was transduced with replication incompetent VSV-g pseudotyped lentiviral particles expressing the firefly luciferase gene under the control of minimal cytomegalovirus (mCMV) promoter and tandem repeats of the TCF/LEF transcriptional response element. Post-transduction selection using puromycin (Sigma Aldrich, cat. no. P8833, 1.5 micrograms per millilitre (ug/mL)) for one week resulted in an enriched polyclonal cell population (DLD1-Wnt-Luc cells) that was expanded and collected for minimal passage and stored in liquid nitrogen.


Wnt-Reporter Assay:

DLD1-Wnt-Luc cells were seeded (5000 cells/well) in a 96-well plate (Greiner Bio-One, cat. no. 655098) in Dubelco's Modified Eagle Medium (DMEM, GIBCO/Invitrogen, cat no. 41965-039) supplemented with Fetal Bovine Serum (FBS, 10%, GIBCO/Invitrogen, cat no. 10108-165). After overnight incubation, the media was replaced with OptiMEM (GIBCO/Invitrogen, cat no. 11058-021) supplemented with FBS (0.5%) and non-essential amino acids (1%, GIBCO/Invitrogen, cat no. 11140-035) and the appropriate putative TNKS inhibitor at a final concentration of 10 μM, 3 μM, 1 μM, 0.30 μM, 0.10 μM, 0.030 μM, 0.010 μM and 0 μM with 1% (v/v) final DMSO in double-triplicate wells. Positive control includes the use of XAV-939 (Maybridge, FisherScientific, 3,5,7,8-tetrahydro-2-[4-(trifluoromethyl)phenyl]-4H-thiopyrano[4,3-c]pyrimidin-4-one, cat. no. RF03920, see: Huang et al., Nature, 2009, Vol. 461, pp. 614-620). Cells were incubated for 20-22 hours before assaying for luciferase (first set of triplicates: Wnt activation) and viability (second set of triplicates: cell survival for data normalisation vs Wnt-activation) using ONE-Glo (Promega, cat. no. E6110) and CellTiter-Glo (Promega, cat. no. G7570) reagents consecutively. The assay was measured using a Victor2 plate reader. The data were normalised to the DMSO control and were expressed as % Wnt activity as a function of inhibitor dose. Dose response curves used to determine IC50 values were Log transformed and analysed by non-linear regression analysis (variable slope) using Prism (GraphPad Software, Inc).


Cell-Based Assay 2:
Western Blotting for Direct and Downstream Targets of TNKS Inhibitors: Axin 1

DLD1 cells were assayed for the effect of putative Tankyrase1/2 inhibitors on TNKS1/2, Axin1/2 and β-catenin protein levels. Effective TNKS inhibitors will (1) increase TNKS protein levels by inhibition of auto-PARylation and subsequent proteasomal degradation, (2) increase Axin1/2 protein levels by inhibition of TNKS induced PARylation and subsequent proteasomal degradation and, consequently, stabilisation of the Axin/APC/GSK/CK destruction complex leading to (3) decrease of β-catenin protein levels. Reduction of nuclear accumulation of β-catenin and ultimately, reduction of β-catenin/TCF/LEF transcriptional activation of Wnt target genes should correlate with loss of Wnt-luc reporter signal in the Wnt reporter assay.


DLD1 cells were seeded in a 6-well dish at 10000 cells/well in DMEM supplemented with 10% FBS. After overnight incubation, cells were dosed with an appropriate amount of putative Tankyrasel/2 inhibitor (2 uM, 0.75 uM, 0.25 uM, 0 uM) in DMEM (0.5% FBS, 1% DMSO). After 20-22 hours incubation, the cells were washed in cold PBS and lysed in lysis buffer (50 mM Tris pH 8.0, 500 mM NaCl, 0.5% NP-40, complete protease inhibitor cocktail (Roche, cat. no. 11836153001)), centrifuged at 15000 rpm for 10 minutes and the protein concentration of the supernatant quantified using Bradford reagent (BioRad protein assay reagent, cat. no. 500-0006). Protein samples (25-50 ug) in protein sample loading buffer (‘Laemmli buffer’, Laemmli, U. K., Nature, 1970, 227, 680-685) were denatured by boiling and loaded onto a sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE with 10% acrylamide) and separated by electrophroresis followed by electroblotting onto nitrocellulose membrane. The membrane was blocked in skimmed milk (5% in Tris-base saline with 0.01% Tween20 (TBS-T)) and subsequently probed overnight with the required antibody: Tankyrasel/2 (1:1000, Santa Cruz, cat. no. sc-8337); Axin1 (1:1000, Cell Signalling Technology, cat. no. 2087); Axin2 (1:1000, Cell Signalling Technology, cat. no. 2151); β-catenin (1:1000, Cell Signalling Technology, cat. no. 9581). After washing in TBS-T, the membrane was probed with a species specific secondary antibody conjugated to HRP (1:5000, Pierce/ThermoFisher), washed again in TBS-T and reacted with chemiluminescent detection reagents (ECL, GE Healthcare, cat. no. RPN2109) followed by exposure to X-ray film (FujiFilm XR).


Cell-Based Assay 3:
Western Blotting for Direct and Downstream Targets of TNKS Inhibitors Unrelated to the Wnt Pathway: TNKS

Appropriate cell lines (HeLa, HT1080, HTC75) were also assayed for the effect of TNKS inhibition on TNKS stabilisation (see, e.g., Smith et al., Science, 1998, Vol. 282, pp. 1484-1487). Cells were seeded, dosed and whole cell lysates were isolated and western blotted as described above. Primary antibodies included TNKS (1:1000, Santa Cruz Biotech, cat. no. SC8377).


Cell-Based Assay 4:
Clonogenic Inhibition in DLD1 or HT55 Cells

In order to determine the efficacy of chronic dosing of putative TNKS inhibitors, long term clonogenic or ‘colony formation’ assays were carried out. This included the sparse seeding of cells in a 6-well dish followed by continuous dosing of cells over 12-14 days (depending on relative cell growth). Appropriate cell lines (DLD1 or HT55) were seeded at 500 cells/well in a 6-well dish in DMEM supplemented with FBS. After overnight incubation, cells were treated with the appropriate putative TN KS inhibitor at 10 μM, 3 μM, 1 μM, 0.30 μM, 0.1 μM and 0 μM at 0.2-1% final DMSO concentration (cell line dependent) in DMEM supplemented with 10% FBS (DLD1 cells were dosed in DMEM supplemented with 0.5% FBS). Dosages were carried out in triplicate. Cell media containing compound or DMSO only was replenished every 48 hours. Termination of the assay included the fixation of cells with trichlororacetic acid (1 mL, 10% (v/v), Sigma Aldrich, cat. no. T6399) and incubation for 16 hours at 4° C. Fixed cells were then washed with water, allowed to dry and stained with sulforhodamine B solution (sulforhodamine B 0.05% (w/v), Sigma Aldrich cat. no. S1402, acetic acid 1% (v/v), Fisher Scientific, cat. no. A/0400/PB17)) for 12 hours at room temperature. The stain was then removed and the cells washed copiously with aqueous acetic acid (1% v/v) and allowed to dry.


Quantification of colony formation was then carried out by dissolution of incorporated sulforhodamine B in Tris-base (1 mL, 10 mM, pH 10) and measurement of absorbance at 560 nM. The data was normalised to the DMSO control and was expressed as surviving fraction as a function of inhibitor dose. Dose response curves used to determine GI50 values were Log transformed and analysed by non-linear regression analysis (variable slope) using Prism (GraphPad Software, Inc).


Biological Data

The following compounds were tested in the TNKS1/PARP Biochemical Assay described above:


IQ-001, IQ-002-1, IQ-002-2, IQ-003, IQ-004, IQ-005, IQ-006, IQ-007, IQ-008, IQ-009, IQ-010, IQ-011, IQ-012, IQ-013, IQ-014, IQ-015, IQ-016, IQ-017, IQ-018, IQ-019, IQ-020, IQ-021, IQ-023, IQ-024, IQ-025, IQ-026, IQ-027, IQ-028-1, IQ-028-2, IQ-029, IQ-030, IQ-031, IQ-032, IQ-033, IQ-034, IQ-035, IQ-036, IQ-037, IQ-038, IQ-039, IQ-040, IQ-041, IQ-042, IQ-043, IQ-044, IQ-045, IQ-046, IQ-047, IQ-048, IQ-049, IQ-050, IQ-051-1, IQ-051-2, IQ-051-3, IQ-052, IQ-053, IQ-054, IQ-055, IQ-056, IQ-057, IQ-059, IQ-060, IQ-062, IQ-063, IQ-065, IQ-067, IQ-068, IQ-070, IQ-071, IQ-072, IQ-073, IQ-074, IQ-075, IQ-076, IQ-077, IQ-078, IQ-079, IQ-080, IQ-081, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-084-3, IQ-085, IQ-086, IQ-087, IQ-088, IQ-089, IQ-090, IQ-091, IQ-092, IQ-093, IQ-094, IQ-095, IQ-096, IQ-097, IQ-098, IQ-099, IQ-100, IQ-101, IQ-102, IQ-103, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-110, IQ-111, IQ-112, IQ-113, IQ-114, IQ-115, IQ-116, IQ-117, IQ-118, IQ-119, IQ-120, IQ-121, IQ-122, IQ-123, IQ-124, IQ-125, IQ-126, IQ-127, IQ-128, IQ-129, IQ-130, IQ-131, IQ-132, IQ-133, IQ-134, IQ-135, IQ-136, IQ-138, IQ-139, IQ-140, IQ-141, IQ-142, IQ-143, IQ-144, IQ-145, IQ-148, IQ-149, IQ-150, IQ-151, IQ-154, IQ-157, IQ-158, IQ-160, IQ-161, IQ-162, IQ-163, IQ-164, IQ-165, IQ-166, IQ-167, IQ-168, IQ-169, IQ-170, IQ-171, IQ-172, IQ-173, IQ-174, IQ-175, IQ-176, IQ-177, IQ-178, IQ-179, IQ-180, IQ-181, IQ-182, IQ-183, IQ-184, IQ-185, IQ-186, IQ-187, IQ-188, IQ-189, IQ-190, IQ-191, IQ-192, IQ-193, IQ-194, IQ-195, IQ-196, IQ-197, IQ-198, IQ-199, IQ-200, IQ-201, IQ-202, IQ-203, IQ-204, IQ-205-1, IQ-205-2, IQ-206, IQ-207-1, IQ-207-2, IQ-208, IQ-209, IQ-210, IQ-211, IQ-212, IQ-213, IQ-214, IQ-215, IQ-216, IQ-217, IQ-218, IQ-219, IQ-220, IQ-221, IQ-222, IQ-223, IQ-224, IQ-225, IQ-226, IQ-227, IQ-228, IQ-229, IQ-230, IQ-231, IQ-232, IQ-233, IQ-234, IQ-236.


All of the compounds have a TNKS1 IC50 of less than 5 μM.


The following compounds have a TNKS1 IC50 of less than 0.5 μM:


IQ-001, IQ-002-1, IQ-002-2, IQ-003, IQ-004, IQ-005, IQ-006, IQ-007, IQ-008, IQ-009, IQ-010, IQ-011, IQ-012, IQ-013, IQ-014, IQ-015, IQ-016, IQ-017, IQ-018, IQ-019, IQ-020, IQ-021, IQ-023, IQ-024, IQ-025, IQ-026, IQ-027, IQ-028-1, IQ-028-2, IQ-029, IQ-031, IQ-032, IQ-033, IQ-034, IQ-035, IQ-036, IQ-037, IQ-038, IQ-039, IQ-040, IQ-041, IQ-042, IQ-043, IQ-044, IQ-045, IQ-046, IQ-047, IQ-048, IQ-049, IQ-050, IQ-051-1, IQ-051-2, IQ-051-3, IQ-052, IQ-053, IQ-054, IQ-055, IQ-056, IQ-057, IQ-059, IQ-060, IQ-062, IQ-063, IQ-065, IQ-067, IQ-068, IQ-070, IQ-071, IQ-072, IQ-073, IQ-074, IQ-075, IQ-076, IQ-077, IQ-078, IQ-079, IQ-080, IQ-081, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-084-3, IQ-085, IQ-086, IQ-087, IQ-088, IQ-089, IQ-090, IQ-091, IQ-092, IQ-093, IQ-094, IQ-095, IQ-096, IQ-097, IQ-098, IQ-099, IQ-100, IQ-101, IQ-102, IQ-103, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-110, IQ-111, IQ-112, IQ-113, IQ-114, IQ-115, IQ-116, IQ-117, IQ-118, IQ-119, IQ-120, IQ-121, IQ-122, IQ-123, IQ-124, IQ-125, IQ-126, IQ-127, IQ-128, IQ-129, IQ-130, IQ-132, IQ-133, IQ-134, IQ-135, IQ-136, IQ-138, IQ-140, IQ-142, IQ-143, IQ-145, IQ-148, IQ-149, IQ-150, IQ-154, IQ-157, IQ-158, IQ-160, IQ-161, IQ-162, IQ-163, IQ-164, IQ-165, IQ-166, IQ-167, IQ-168, IQ-169, IQ-170, IQ-171, IQ-172, IQ-173, IQ-174, IQ-175, IQ-176, IQ-177, IQ-178, IQ-179, IQ-180, IQ-181, IQ-182, IQ-184, IQ-185, IQ-186, IQ-187, IQ-188, IQ-189, IQ-190, IQ-191, IQ-192, IQ-193, IQ-194, IQ-195, IQ-196, IQ-197, IQ-198, IQ-199, IQ-200, IQ-201, IQ-202, IQ-203, IQ-204, IQ-205-1, IQ-205-2, IQ-206, IQ-207-1, IQ-207-2, IQ-208, IQ-209, IQ-210, IQ-211, IQ-212, IQ-213, IQ-214, IQ-215, IQ-216, IQ-217, IQ-218, IQ-219, IQ-220, IQ-221, IQ-222, IQ-223, IQ-224, IQ-225, IQ-226, IQ-227, IQ-228, IQ-229, IQ-230, IQ-231, IQ-232, IQ-234, IQ-236.


The following compounds have a TNKS1 IC50 of less than 0.05 μM:


IQ-001, IQ-002-1, IQ-003, IQ-004, IQ-005, IQ-006, IQ-007, IQ-008, IQ-011, IQ-012, IQ-013, IQ-014, IQ-015, IQ-016, IQ-017, IQ-018, IQ-019, IQ-021, IQ-023, IQ-025, IQ-026, IQ-027, IQ-028-1, IQ-028-2, IQ-029, IQ-032, IQ-033, IQ-034, IQ-035, IQ-036, IQ-037, IQ-038, IQ-042, IQ-045, IQ-048, IQ-050, IQ-051-1, IQ-051-2, IQ-051-3, IQ-052, IQ-053, IQ-054, IQ-055, IQ-056, IQ-057, IQ-059, IQ-062, IQ-065, IQ-067, IQ-068, IQ-070, IQ-073, IQ-074, IQ-078, IQ-080, IQ-081, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-084-3, IQ-085, IQ-086, IQ-087, IQ-088, IQ-090, IQ-093, IQ-094, IQ-095, IQ-096, IQ-097, IQ-098, IQ-099, IQ-100, IQ-101, IQ-102, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-111, IQ-112, IQ-115, IQ-116, IQ-117, IQ-118, IQ-120, IQ-121, IQ-123, IQ-124, IQ-125, IQ-127, IQ-129, IQ-130, IQ-134, IQ-138, IQ-149, IQ-158, IQ-160, IQ-162, IQ-167, IQ-168, IQ-169, IQ-170, IQ-171, IQ-172, IQ-173, IQ-174, IQ-175, IQ-176, IQ-177, IQ-178, IQ-180, IQ-182, IQ-185, IQ-187, IQ-188, IQ-189, IQ-190, IQ-191, IQ-192, IQ-193, IQ-194, IQ-195, IQ-196, IQ-197, IQ-198, IQ-199, IQ-200, IQ-201, IQ-203, IQ-204, IQ-205-1, IQ-205-2, IQ-206, IQ-207-1, IQ-207-2, IQ-208, IQ-209, IQ-210, IQ-211, IQ-212, IQ-213, IQ-214, IQ-215, IQ-216, IQ-217, IQ-218, IQ-219, IQ-220, IQ-222, IQ-224, IQ-226, IQ-227, IQ-228, IQ-231.


The following compounds have a TNKS1 IC50 of less than 0.02 μM:


IQ-001, IQ-004, IQ-005, IQ-006, IQ-008, IQ-011, IQ-014, IQ-016, IQ-017, IQ-018, IQ-025, IQ-028-1, IQ-029, IQ-032, IQ-034, IQ-038, IQ-048, IQ-051-1, IQ-054, IQ-055, IQ-062, IQ-082, IQ-086, IQ-088, IQ-093, IQ-097, IQ-099, IQ-100, IQ-102, IQ-104, IQ-107, IQ-109, IQ-115, IQ-117, IQ-118, IQ-120, IQ-123, IQ-125, IQ-130, IQ-162, IQ-167, IQ-168, IQ-170, IQ-172, IQ-175, IQ-176, IQ-177, IQ-178, IQ-180, IQ-182, IQ-188, IQ-189, IQ-190, IQ-191, IQ-192, IQ-193, IQ-194, IQ-195, IQ-196, IQ-197, IQ-198, IQ-199, IQ-200, IQ-204, IQ-205-2, IQ-208, IQ-209, IQ-210, IQ-211, IQ-213, IQ-214, IQ-215, IQ-219, IQ-222, IQ-227.


For example, IQ-016 has a TNKS1 IC50 of 0.012 μM.


The following compounds were tested in the Wnt-Luciferase Reporter Assay described above:


IQ-001, IQ-002-1, IQ-003, IQ-004, IQ-005, IQ-006, IQ-007, IQ-008, IQ-009, IQ-010, IQ-011, IQ-012, IQ-013, IQ-014, IQ-015, IQ-016, IQ-017, IQ-018, IQ-019, IQ-020, IQ-021, IQ-023, IQ-024, IQ-025, IQ-026, IQ-027, IQ-028-1, IQ-029, IQ-031, IQ-032, IQ-033, IQ-034, IQ-035, IQ-036, IQ-037, IQ-038, IQ-040, IQ-041, IQ-042, IQ-043, IQ-045, IQ-046, IQ-048, IQ-050, IQ-051-1, IQ-051-2, IQ-051-3, IQ-052, IQ-053, IQ-054, IQ-055, IQ-056, IQ-057, IQ-059, IQ-060, IQ-062, IQ-063, IQ-065, IQ-067, IQ-068, IQ-070, IQ-071, IQ-072, IQ-073, IQ-074, IQ-075, IQ-076, IQ-077, IQ-078, IQ-079, IQ-080, IQ-081, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-084-3, IQ-085, IQ-086, IQ-087, IQ-088, IQ-089, IQ-090, IQ-091, IQ-093, IQ-094, IQ-095, IQ-096, IQ-097, IQ-098, IQ-099, IQ-100, IQ-101, IQ-102, IQ-103, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-110, IQ-111, IQ-112, IQ-115, IQ-116, IQ-117, IQ-118, IQ-119, IQ-120, IQ-121, IQ-122, IQ-123, IQ-124, IQ-125, IQ-127, IQ-128, IQ-129, IQ-130, IQ-132, IQ-133, IQ-134, IQ-135, IQ-138, IQ-142, IQ-143, IQ-148, IQ-149, IQ-150, IQ-151, IQ-154, IQ-157, IQ-158, IQ-160, IQ-161, IQ-162, IQ-163, IQ-164, IQ-165, IQ-166, IQ-167, IQ-168, IQ-169, IQ-170, IQ-171, IQ-172, IQ-173, IQ-174, IQ-175, IQ-176, IQ-177, IQ-178, IQ-179, IQ-180, IQ-181, IQ-182, IQ-183, IQ-184, IQ-185, IQ-186, IQ-187, IQ-188, IQ-189, IQ-190, IQ-191, IQ-192, IQ-193, IQ-194, IQ-195, IQ-196, IQ-197, IQ-198, IQ-199, IQ-200, IQ-201, IQ-202, IQ-203, IQ-204, IQ-205-1, IQ-205-2, IQ-206, IQ-207-1, IQ-207-2, IQ-208, IQ-209, IQ-210, IQ-211, IQ-212, IQ-213, IQ-214, IQ-215, IQ-216, IQ-217, IQ-218, IQ-219, IQ-220, IQ-221, IQ-222, IQ-223, IQ-224, IQ-225, IQ-226, IQ-227, IQ-228, IQ-229, IQ-230, IQ-231, IQ-232, IQ-234, IQ-236.


All of the compounds have a Wnt IC50 of less than 10 μM.


The following compounds have a Wnt IC50 of less than 5 μM:


IQ-001, IQ-002-1, IQ-003, IQ-004, IQ-005, IQ-006, IQ-007, IQ-008, IQ-009, IQ-010, IQ-011, IQ-012, IQ-013, IQ-014, IQ-015, IQ-016, IQ-017, IQ-018, IQ-019, IQ-020, IQ-021, IQ-023, IQ-024, IQ-025, IQ-026, IQ-027, IQ-028-1, IQ-029, IQ-031, IQ-032, IQ-033, IQ-034, IQ-035, IQ-036, IQ-037, IQ-038, IQ-040, IQ-041, IQ-042, IQ-043, IQ-045, IQ-046, IQ-048, IQ-050, IQ-051-1, IQ-051-2, IQ-051-3, IQ-052, IQ-053, IQ-054, IQ-055, IQ-056, IQ-057, IQ-059, IQ-060, IQ-062, IQ-063, IQ-065, IQ-067, IQ-068, IQ-070, IQ-071, IQ-072, IQ-073, IQ-074, IQ-075, IQ-076, IQ-077, IQ-078, IQ-079, IQ-080, IQ-081, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-084-3, IQ-085, IQ-086, IQ-087, IQ-088, IQ-089, IQ-090, IQ-091, IQ-093, IQ-094, IQ-095, IQ-096, IQ-097, IQ-098, IQ-099, IQ-100, IQ-101, IQ-102, IQ-103, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-110, IQ-111, IQ-112, IQ-115, IQ-116, IQ-117, IQ-118, IQ-119, IQ-120, IQ-121, IQ-122, IQ-123, IQ-124, IQ-125, IQ-127, IQ-128, IQ-129, IQ-130, IQ-132, IQ-133, IQ-134, IQ-135, IQ-138, IQ-142, IQ-143, IQ-148, IQ-149, IQ-150, IQ-151, IQ-154, IQ-157, IQ-158, IQ-160, IQ-161, IQ-162, IQ-163, IQ-164, IQ-165, IQ-166, IQ-167, IQ-168, IQ-169, IQ-170, IQ-171, IQ-172, IQ-173, IQ-174, IQ-175, IQ-176, IQ-177, IQ-178, IQ-179, IQ-180, IQ-181, IQ-182, IQ-183, IQ-184, IQ-185, IQ-186, IQ-187, IQ-188, IQ-189, IQ-190, IQ-191, IQ-192, IQ-193, IQ-194, IQ-195, IQ-196, IQ-197, IQ-198, IQ-199, IQ-200, IQ-201, IQ-202, IQ-203, IQ-204, IQ-205-1, IQ-205-2, IQ-206, IQ-207-1, IQ-207-2, IQ-208, IQ-209, IQ-210, IQ-211, IQ-212, IQ-213, IQ-214, IQ-215, IQ-216, IQ-217, IQ-218, IQ-219, IQ-220, IQ-221, IQ-222, IQ-223, IQ-224, IQ-225, IQ-226, IQ-227, IQ-228, IQ-230, IQ-231, IQ-232, IQ-234.


The following compounds have a Wnt IC50 of less than 0.5 μM:


IQ-001, IQ-002-1, IQ-003, IQ-004, IQ-005, IQ-006, IQ-007, IQ-008, IQ-009, IQ-010, IQ-011, IQ-012, IQ-013, IQ-014, IQ-015, IQ-016, IQ-017, IQ-018, IQ-019, IQ-020, IQ-021, IQ-023, IQ-025, IQ-026, IQ-027, IQ-028-1, IQ-029, IQ-031, IQ-034, IQ-035, IQ-036, IQ-037, IQ-038, IQ-040, IQ-041, IQ-042, IQ-043, IQ-045, IQ-048, IQ-050, IQ-051-1, IQ-051-2, IQ-051-3, IQ-052, IQ-053, IQ-054, IQ-055, IQ-056, IQ-057, IQ-059, IQ-060, IQ-062, IQ-063, IQ-065, IQ-067, IQ-068, IQ-071, IQ-072, IQ-073, IQ-074, IQ-075, IQ-076, IQ-077, IQ-078, IQ-079, IQ-080, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-084-3, IQ-085, IQ-086, IQ-087, IQ-088, IQ-089, IQ-090, IQ-091, IQ-095, IQ-096, IQ-097, IQ-098, IQ-099, IQ-100, IQ-101, IQ-102, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-110, IQ-111, IQ-112, IQ-115, IQ-116, IQ-117, IQ-118, IQ-119, IQ-120, IQ-121, IQ-122, IQ-123, IQ-125, IQ-127, IQ-130, IQ-133, IQ-134, IQ-138, IQ-142, IQ-143, IQ-148, IQ-154, IQ-157, IQ-158, IQ-161, IQ-162, IQ-166, IQ-167, IQ-168, IQ-169, IQ-170, IQ-171, IQ-172, IQ-173, IQ-174, IQ-175, IQ-176, IQ-177, IQ-178, IQ-179, IQ-180, IQ-181, IQ-182, IQ-183, IQ-184, IQ-185, IQ-186, IQ-187, IQ-188, IQ-189, IQ-190, IQ-192, IQ-193, IQ-194, IQ-195, IQ-196, IQ-197, IQ-198, IQ-199, IQ-200, IQ-201, IQ-202, IQ-203, IQ-204, IQ-205-1, IQ-205-2, IQ-206, IQ-207-1, IQ-207-2, IQ-208, IQ-209, IQ-210, IQ-211, IQ-212, IQ-213, IQ-214, IQ-215, IQ-218, IQ-219, IQ-220, IQ-222, IQ-226, IQ-227, IQ-228, IQ-231, IQ-234.


The following compounds have a Wnt IC50 of less than 0.05 μM:


IQ-001, IQ-003, IQ-004, IQ-005, IQ-006, IQ-008, IQ-011, IQ-015, IQ-016, IQ-017, IQ-018, IQ-028-1, IQ-035, IQ-038, IQ-040, IQ-042, IQ-048, IQ-051-2, IQ-051-3, IQ-054, IQ-055, IQ-062, IQ-065, IQ-067, IQ-068, IQ-073, IQ-078, IQ-080, IQ-097, IQ-098, IQ-100, IQ-102, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-111, IQ-117, IQ-118, IQ-120, IQ-121, IQ-123, IQ-125, IQ-133, IQ-148, IQ-157, IQ-167, IQ-168, IQ-170, IQ-171, IQ-173, IQ-174, IQ-175, IQ-176, IQ-177, IQ-178, IQ-179, IQ-180, IQ-181, IQ-182, IQ-184, IQ-185, IQ-190, IQ-192, IQ-195, IQ-198, IQ-201, IQ-206, IQ-209, IQ-210, IQ-211, IQ-212, IQ-215, IQ-234.


For example, IQ-016 has a Wnt IC50 of 0.014 μM.


The following compounds were studied using the Western Blotting Assays described above, and were found to stabilize Axin1 and to stabilize TNKS: IQ-002-1, IQ-003, IQ-027, IQ-034, IQ-036, IQ-037, IQ-038, IQ-053, IQ-100, IQ-102, IQ-127, IQ-130, IQ-133.


The following compounds were tested in the Long-Term Clonogenic Assay described above (DLD1 cells):


IQ-001, IQ-002-1, IQ-003, IQ-004, IQ-005, IQ-006, IQ-007, IQ-008, IQ-009, IQ-011, IQ-016, IQ-017, IQ-018, IQ-019, IQ-021, IQ-023, IQ-026, IQ-027, IQ-028-1, IQ-032, IQ-034, IQ-038, IQ-040, IQ-042, IQ-043, IQ-048, IQ-051-2, IQ-051-3, IQ-053, IQ-054, IQ-057, IQ-065, IQ-067, IQ-068, IQ-072, IQ-073, IQ-074, IQ-075, IQ-081, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-086, IQ-088, IQ-090, IQ-091, IQ-095, IQ-096, IQ-097, IQ-099, IQ-100, IQ-101, IQ-102, IQ-103, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-111, IQ-118, IQ-121, IQ-123, IQ-125, IQ-127, IQ-128, IQ-129, IQ-130, IQ-149, IQ-161, IQ-162, IQ-166, IQ-167, IQ-168, IQ-169, IQ-188, IQ-189, IQ-190, IQ-231, IQ-234.


All of the compounds have a Clonogenic SF50 (DLD1) of less than 10 μM.


The following compounds have a Clonogenic SF50 (DLD1) of less than 2 μM:


IQ-001, IQ-002-1, IQ-003, IQ-005, IQ-006, IQ-007, IQ-008, IQ-009, IQ-011, IQ-016, IQ-017, IQ-018, IQ-019, IQ-021, IQ-023, IQ-027, IQ-028-1, IQ-034, IQ-038, IQ-040, IQ-042, IQ-043, IQ-048, IQ-051-2, IQ-051-3, IQ-053, IQ-054, IQ-057, IQ-065, IQ-067, IQ-068, IQ-073, IQ-074, IQ-075, IQ-081, IQ-082, IQ-083, IQ-084-1, IQ-084-2, IQ-086, IQ-088, IQ-090, IQ-091, IQ-102, IQ-104, IQ-105, IQ-106, IQ-107, IQ-108, IQ-109, IQ-111, IQ-118, IQ-121, IQ-123, IQ-125, IQ-127, IQ-129, IQ-149, IQ-161, IQ-162, IQ-166, IQ-168, IQ-190.


The following compounds have a Clonogenic SF50 (DLD1) of less than 0.5 μM:


IQ-006, IQ-007, IQ-008, IQ-011, IQ-016, IQ-018, IQ-027, IQ-028-1, IQ-040, IQ-042, IQ-051-2, IQ-053, IQ-065, IQ-067, IQ-073, IQ-075, IQ-081, IQ-082, IQ-083, IQ-088, IQ-090, IQ-091, IQ-104, IQ-105, IQ-107, IQ-108, IQ-109, IQ-111, IQ-118, IQ-123, IQ-125, IQ-161, IQ-162, IQ-166, IQ-168.


For example, IQ-016 has a Clonogenic SF50 (DLD1) of 0.291 μM.


The following compounds were tested in the Long-Term Clonogenic Assay described above (HT55 cells):


IQ-168, IQ-185, IQ-007, IQ-018, IQ-027, IQ-053, IQ-173, IQ-006, IQ-195, IQ-075, IQ-080, IQ-170, IQ-016, IQ-011, IQ-182, IQ-174, IQ-177, IQ-178, IQ-197, IQ-201, IQ-158, IQ-204, IQ-048, IQ-196, IQ-117, IQ-210, IQ-199, IQ-176, IQ-059, IQ-179, IQ-198, IQ-054, IQ-209, IQ-005, IQ-042, IQ-213, IQ-218, IQ-100, IQ-127, IQ-171, IQ-208, IQ-206, IQ-205-1, IQ-205-2, IQ-207-1, IQ-207-2, IQ-028-1.


All of the compounds have a Clonogenic SF50 (HT55) of less than 10 μM.


The following compounds have a Clonogenic SF50 (HT55) of less than 3 μM:


IQ-006, IQ-007, IQ-011, IQ-016, IQ-018, IQ-027, IQ-028-1, IQ-048, IQ-053, IQ-059, IQ-075, IQ-080, IQ-117, IQ-158, IQ-168, IQ-170, IQ-173, IQ-174, IQ-176, IQ-177, IQ-178, IQ-179, IQ-182, IQ-185, IQ-195, IQ-196, IQ-197, IQ-199, IQ-201, IQ-204, IQ-205-1, IQ-205-2, IQ-207-1, IQ-207-2, IQ-210.


The following compounds have a Clonogenic SF50 (HT55) of less than 1.5 μM:


IQ-006, IQ-007, IQ-011, IQ-016, IQ-018, IQ-027, IQ-028-1, IQ-053, IQ-075, IQ-080, IQ-168, IQ-170, IQ-173, IQ-185, IQ-195, IQ-205-1, IQ-207-1.

For example, IQ-016 has a Clonogenic SF50 (HT55) of 1.235 μM.


—R5Comparison No. 1:

As demonstrated by this comparison, the presence of R5 as -Me (as compared to —H) decreased Wnt IC50 by a factor of about 13.

















TNKS1
Wnt


Code
Structure
IC50 (μM)
IC50 (μM)







IQ-025


embedded image


0.017
0.062





REF-1


embedded image


0.033
0.825









—R5Comparison No. 2:

As demonstrated by this comparison, the presence of R5 as -Me (as compared to —H) decreased Wnt IC50 (by a factor of about 24).

















TNKS1
Wnt




IC50
IC50


Code
Structure
(μM)
(μM)







IQ-080


embedded image


0.029
0.042





REF-2


embedded image


0.048
1.003









—R5Comparison No. 3:

As demonstrated by this comparison, the presence of R5 as -Me (as compared to —H) decreased Wnt IC50 (by a factor of about 62).


Also as demonstrated by this comparison, the presence of R5 as —Cl (as compared to —H) decreased Wnt IC50 (by a factor of about 4).

















TNKS1
Wnt


Code
Structure
IC50 (μM)
IC50 (μM)







IQ-003


embedded image


0.021
0.012





IQ-002


embedded image


0.039
0.179





REF-3


embedded image


0.024
0.742









—R5Comparison No. 4:

As demonstrated by this comparison, the presence of R5 as -Me (as compared to —H) decreased Wnt IC50 (by a factor of at least 9).

















TNKS1
Wnt




IC50
IC50


Code
Structure
(μM)
(μM)







IQ-034


embedded image


0.012
1.07





REF-4


embedded image


0.018
>10  









—R5Comparison No. 5:

As demonstrated by this comparison, the presence of R5 as -Me (as compared to —H) decreased Wnt IC50 (by a factor of at least 60).

















TNKS1
Wnt




IC50
IC50


Code
Structure
(μM)
(μM)







IQ-130


embedded image


0.016
0.165





REF-5


embedded image


0.017
>10   









—R5Comparison No. 6:

As demonstrated by this comparison, the presence of R5 as -Me (as compared to —OH) decreased Wnt IC50 (by a factor of at least 14).

















TNKS1
Wnt




IC50
IC50


Code
Structure
(μM)
(μM)







IQ-157


embedded image


0.051
0.041





REF-6


embedded image


0.024
0.611









—R5Comparison No. 7:

As demonstrated by this comparison, the presence of R5 as -Me (as compared to —OH) decreased Wnt IC50 (by a factor of at least 36).


Also as demonstrated by this comparison, the additional change of R7 as —F (as compared to —H) further decreased Wnt IC50 (now by a factor of at least 60).

















TNKS1
Wnt




IC50
IC50


Code
Structure
(μM)
(μM)







IQ-220


embedded image


0.026
0.274





IQ-222


embedded image


0.016
0.174





REF-7


embedded image


0.042
>10   









-L3P-R3NComparison No. 1:

As demonstrated by this comparison, the presence of -L3P-R3N as N-(cyclopropylmethyl)-piperazino-carbonyl (as compared to —OMe) decreased Wnt IC50 (by a factor of at least about 3).

















TNKS1
Wnt




IC50
IC50


Code
Structure
(μM)
(μM)







IQ-223


embedded image


0.076
1.34





REF-8


embedded image


0.039
4.32









The foregoing has described the principles, preferred embodiments, and modes of operation of the present invention. However, the invention should not be construed as limited to the particular embodiments discussed. Instead, the above-described embodiments should be regarded as illustrative rather than restrictive. It should be appreciated that variations may be made in those embodiments by workers skilled in the art without departing from the scope of the present invention.


REFERENCES

A number of publications are cited herein in order to more fully describe and disclose the invention and the state of the art to which the invention pertains. Full citations for these references are provided below. Each of these references is incorporated herein by reference in its entirety into the present disclosure, to the same extent as if each individual reference was specifically and individually indicated to be incorporated by reference.

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Claims
  • 1-137. (canceled)
  • 138. A compound selected from compounds of the following formula, and pharmaceutically acceptable salts and N-oxides thereof:
  • 139. A compound according to claim 138, wherein: W is CRW, X is CRX, Y is CRY, and Z is CRZ (“phenyl”).
  • 140. A compound according to claim 138, wherein: W is CRW, X is N, Y is CRY, and Z is CRZ (“pyrid-3-yl”).
  • 141. A compound according to claim 138, wherein: —RW, if present, is —H;—RX, if present, is —H;—RY, if present, is —H; and—RZ, if present, is —H.
  • 142. A compound according to claim 138, wherein -L3P- is a single covalent bond.
  • 143. A compound according to claim 138, wherein -L3P- is -L3PL-.
  • 144. A compound according to claim 138, wherein -L3PL-, if present, is -L3PR1-.
  • 145. A compound according to claim 138, wherein -L3PL-, if present, is —C(═O)—.
  • 146. A compound according to claim 138, wherein -L3PL-, if present, is -L3PR2-C(═O)—.
  • 147. A compound according to claim 138, wherein: each -L3PR1-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-; andeach -L3PR2-, if present, is independently —CH2—, —CH(Me)—, or —C(Me)2-.
  • 148. A compound according to claim 138, wherein —R3N is —NRARB.
  • 149. A compound according to claim 138, wherein —R3N is —NRCRD.
  • 150. A compound according to claim 138, wherein each —RA, if present, is independently: —RA1, —RA3, or -LA-RA3.
  • 151. A compound according to claim 138, wherein: each —RA1, if present, is independently linear or branched saturated C1-4alkyl, and is optionally substituted with one or more groups —RS1; andeach —RA3, if present, is piperidinyl,
  • 152. A compound according to claim 138, wherein each —RSN, if present, is independently: —RTT,—C(═O)RTT, or—C(═O)ORTT.
  • 153. A compound according to claim 138, wherein each —RTT, if present, is -Me.
  • 154. A compound according to claim 138, wherein —NRCRD, if present, is —NRC1RD1 wherein, —NRC1RD1 is independently selected from the following groups, and is: optionally substituted on carbon with one or more groups —RNC, andoptionally substituted on secondary nitrogen, if present, with a group —RNN:
  • 155. A compound according to claim 138, wherein: each —RNC, if present, is independently: —RQQ,—OH, —ORQQ,—NH2, —NHRQQ, —NRQQ2, —RQM, or═O;each —RNN, if present, is independently: —RQQ,-LQ-OH, -LQ-ORQQ,-LQ-NH2, -LQ-NHRQQ, -LQ-NRQQ2, -LQ-RQM,—C(═O)RQQ, or—C(═O)ORQQ; andeach —RQQ is independently linear or branched saturated C1-4alkyl, saturated C3-6cycloalkyl, or saturated C3-6cycloalkyl-methyl.
  • 156. A compound according to claim 138, wherein each —RQQ, if present, is -Me.
  • 157. A compound according to claim 138, wherein —NRCRD, if present, is —NRC5RD5, wherein —NRC5RD5 is: 1H-pyrazol-1-yl; and is optionally substituted with one or more groups —RH.
  • 158. A compound according to claim 138, wherein —NRCRD, if present, is —NRC5RD5, wherein —NRC5RD5 is: 1H-imidazol-1-yl; and is optionally substituted with one or more groups —RH.
  • 159. A compound according to claim 138, wherein each —RH, if present, is independently —RHH.
  • 160. A compound according to claim 138, wherein —R5 is —R5A, wherein —R5A is -Me.
  • 161. A compound according to claim 138, wherein —R5 is —R5C, wherein —R5C is —Cl.
  • 162. A compound according to claim 138, wherein —R6 is —H.
  • 163. A compound according to claim 138, selected from compounds of the following formulae and pharmaceutically acceptable salts and N-oxides thereof: IQ-001 through IQ-238.
  • 164. A pharmaceutical composition comprising a compound according to claim 138, and a pharmaceutically acceptable carrier or diluent.
  • 165. A method of preparing a pharmaceutical composition comprising the step of mixing a compound according to claim 138, and a pharmaceutically acceptable carrier or diluent.
  • 166. A method of inhibiting PARP function, TNKS1 and/or TNKS2 function, or Wnt signalling in a cell, in vitro or in vivo, comprising contacting the cell with an effective amount of a compound according to claim 138.
  • 167. A method of treatment of a proliferative condition, comprising administering to a subject in need of treatment a therapeutically-effective amount of a compound according to claim 138.
PCT Information
Filing Document Filing Date Country Kind
PCT/GB2013/050561 3/7/2013 WO 00
Provisional Applications (1)
Number Date Country
61607680 Mar 2012 US