Patent Application
20100168170
The present invention is directed to substituted lactam compounds and their uses as therapeutic agents.
Inflammation is an essential localized host response to invading microorganisms or tissue injury which involves cells of the immune system. The classic signs of inflammation include redness (erythema), swelling (edema), pain and increased heat production (pyrema) at the site of injury. The inflammatory response allows the body to specifically recognize and eliminate an invading organism and/or repair tissue injury. Many of the acute changes at the site of inflammation are either directly or indirectly attributable to the massive influx of leukocytes (e.g., neutrophils, eosinophils, lymphocytes, monocytes) which is intrinsic to this response. Leukocytic infiltration and accumulation in tissue results in their activation and subsequent release of inflammatory mediators such as LTB4, prostaglandins, TNF-α, IL-β, IL-8, IL-5, IL-6, histamine, proteases and reactive oxygen species for example.
Normal inflammation is a highly regulated process that is tightly controlled at several levels for each of the cell types involved in the response. For example, expression of the pro-inflammatory cytokine TNF-α is controlled at the level of gene expression, translation, post-translational modification and release of the mature form from the cell membrane. Many of the proteins up-regulated during inflammation are controlled by the transcription factor, NF-κB. Pro-inflammatory responses are normally countered by endogenous anti-inflammatory mechanisms such as generation of IL-10 or IL-4. A characteristic of a normal inflammatory response is that it is temporary in nature and is followed by a resolution phase which brings the state of the tissue back to its prior condition. The resolution phase is thought to involve up-regulation of anti-inflammatory mechanisms, such as IL-10, as well as down-regulation of the proinflammatory processes.
Inflammatory disease occurs when an inflammatory response is initiated that is inappropriate and/or does not resolve in the normal manner but rather persists and results in a chronic inflammatory state. Inflammatory disease may be systemic (e.g. lupus) or localized to particular tissues or organs and exerts an enormous personal and economic burden on society. Examples of some of the most common and problematic inflammatory diseases are rheumatoid arthritis, inflammatory bowel disease, psoriasis, asthma, chronic obstructive pulmonary disease, emphysema, colitis and ischemia-reperfusion injury.
A common underlying theme in inflammatory disease is a perturbation of the cellular immune response that results in recognition of host proteins (antigens) as foreign. Thus the inflammatory response becomes misdirected at host tissues with effector cells targeting specific organs or tissues often resulting in irreversible damage. The self-recognition aspect of auto-immune disease is often reflected by the clonal expansion of T-cell subsets characterized by a particular T-cell receptor (TCR) subtype in the disease state. Often inflammatory disease is also characterized by an imbalance in the levels of T-helper (Th) subsets (i.e., Th1 cells vs. Th2 cells).
Therapeutic strategies aimed at curing inflammatory diseases usually fall into one of two categories: (a) down-modulation of processes that are up-regulated in the disease state or (b) up-regulation of anti-inflammatory pathways in the affected cells or tissues. Most regimes currently employed in the clinic fall into the first category. Some examples of which are corticosteroids and non-steroidal anti-inflammatory drugs (NSAIDs).
Many of the tissue, cellular and biochemical processes which are perturbed in inflammatory disease have been elucidated and this has allowed the development of experimental models or assays to mimic the disease state. These in-vitro assays enable selection and screening of compounds with a high probability of therapeutic efficacy in the relevant inflammatory disease. Thus, currently employed assays used to model the importance of the activated leukocytes in the development of acute inflammation and maintenance of the chronic inflammatory state are assays monitoring leukocyte chemotaxis and cellular degranulation and cytokine synthesis and reactive oxygen species (ROS) production assays in vitro. Since a result of acute or chronic neutrophil activation is release of ROS with resultant tissue damage, an assay for scavengers of ROS allows detection of compounds with potential therapeutic efficacy.
Cellular assays to detect inhibitors of TNF-α release from stimulated macrophage or monocytic cells are an important component of an in vitro model for inflammation as this cytokine is upregulated and has been shown to contribute to the pathology in many inflammatory diseases. Since elevated cAMP in affected cells has been shown to modulate or dampen the inflammatory response, monitoring cellular cyclic AMP (cAMP) levels, and the activity of pathways controlling cAMP levels allows for the detection of potential anti-inflammatory compounds. Assays may include monitoring the level of cAMP itself, phosphodiesterase activity, or changes in cAMP response element (CRE)-luciferase activity.
The cyclic nucleotides, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), play a key role in regulating cell function and phosphodiesterases (PDEs) provide the main route for the degradation of cyclic nucleotides. cAMP is now known to control the functional and genomic responses for a variety of cellular functions triggered by a wide array of receptors (Beavo, J. A. and Brunton, L. L., Nat. Rev. Mol. Cell Biol., 3, 710-718 (2002)). Local control of cAMP signalling is affected by a complex pattern of localized synthesis, by adenylate cyclise (AC), and by phosphodiesterase (PDE)-mediated enzymatic degradation.
The PDEs are a family of enzymes that catalyze the hydrolysis of 3′,5′-cyclic nucleotides to 5′ nucleoside monophosphates, including the conversion of cAMP to AMP and cGMP to GMP. PDE enzymes are collectively grouped as a superfamily of eleven different, but homologous, gene-families with a highly conserved catalytic domain (Soderling, S. H. and Beavo, J. A., Curr. Opin. Cell Biol., 12, 174-179 (2000)). At present twenty-one different mammalian PDE genes have been identified. Many of these genes are expressed in multiple isoforms either by differing initiation sequences or splicing patterns. Differentiation of the enzymes can be achieved on the basis of substrate specificity, kinetic properties and sensitivity to regulatory molecules. PDEs in families 5, 6 and 9 specifically catalyze the hydrolysis of cGMP while PDEs 4, 7 and 8 are specific for cAMP. Enzymes belonging to the other PDE families (1, 2, 3, 10 and 11) catalyze the hydrolysis of both cAMP and cGMP with differing kinetics. Different PDE isozymes can have specific tissue, cellular and subcellular distributions and more than one type of PDE is usually present in any given cell. The types of PDEs expressed in a cell, together with their relative proportions and subcellular localization, control the cyclic nucleotide phenotype of that cell.
The PDE4 enzyme is responsible for selective, high affinity hydrolytic degradation of the second messenger cAMP, has a low Michaelis constant and is sensitive to inhibition by rolipram. The PDE4 enzyme family consists of four genes, which produce 4 isoforms of the PDE4 enzyme (PDE4A, PDE4B, PDE4C, and PDE4D) (Wang et al., “Expression, Purification, and Characterization of human cAMPSpecific Phosphodiesterase (PDE4) Subtypes A, B, C, and D, Biochem”, Biophys. Res. Comm., 234, 320-324 (1997)). Moreover, various splice variants of each PDE4 isoform have been identified and play a role in the compartmentalized cAMP signalling in cells (Houslay, M. D., Schafer, P., and Zhang, K. Y., Drug Discov. Today, 15; 10(22):1503-19 (2005)). Recently, a number of selective PDE4 inhibitors have been discovered to have beneficial pharmacological effects resulting from PDE4 inhibition as shown in a variety of disease models (Torphy et al., Environ. Health Perspect., 102 Suppl. 10, 79-84, 1994; Duplantier et al., J. Med. Chem., 39 120-125 (1996); Schneider et al., Pharmacol. Biochem. Behav., 50, 211-217 (1995); Banner and Page, Br. J. Pharmacol., 114, 93-98 (1995); Barnette et al., J. Pharmacol. Exp. Ther., 273, 674-679 (1995); Wright et al., “Differential in vivo and in vitro bronchorelaxant activities of CP-80633, a selective phosphodiesterase 4 inhibitor,” Can. J. Physiol. Pharmacol., 75, 1001-1008 (1997); Manabe et al., “Anti-inflammatory and bronchodilator properties of KF19514, a phosphodiesterase 4 and 1 inhibitor,” Eur. J. Pharmacol., 332, 97-107 (1997); and Ukita et al., “Novel, potent, and selective phosphodiesterase-4 inhibitors as antiasthmatic agents: synthesis and biological activities of a series of 1-pyridylnaphthalene derivatives,” J. Med. Chem., 42, 1088-1099 (1999)). Therefore, considerable interest exists in the discovery of additional selective inhibitors of PDE4.
Regulation of cAMP activity is important in many biological processes, including inflammation, depression and cognitive function. Chronic inflammation is a multitude of heterogeneous diseases characterized in part by activation of multiple inflammatory cells, particularly cells of lymphoid lineage (including T lymphocytes) and myeloid lineage (including granulocytes, macrophages, and monocytes). Activation of these inflammatory cells results in production and release of proinflammatory mediators, including cytokines and chemokines, such as tumor necrosis factor (TNF) and interleukin-1 (IL-1). Discovery of a molecule that suppresses or inhibits such cellular activation and proinflammatory mediator release would be useful in the therapeutic treatment of inflammatory diseases. Elevated cAMP levels suppress inflammatory cell activation. Increased cAMP levels associated with PDE4 inhibition has therefore become a valid potential therapeutic approach to control inflammatory responses and disorders (Beavo et al., “Cyclic Nucleotide Phosphodiesterases: Structure, Regulation and Drug Action,” Wiley and Sons, Chichester, pp. 3-14 (1990); Torphy et al., Drug News and Perspectives, 6, pp. 203-214 (1993); Giembycz et al., Clin. Exp. Allergy, 22, pp. 337-344 (1992); and Sanz, M. J., Cortijo, J., Morcillo, E. J., Pharmacol Ther. 106(3):269-97 (2005)).
PDE4 inhibitors have recently shown clinical utility in mitigating the effects of the chronic pulmonary inflammatory diseases of asthma and chronic obstructive pulmonary disease (COPD). Roflumilast, a selective PDE4 inhibitor, demonstrated improvements in measures of airway function (forced expiratory volume in 1 second; FEV1, and peak expiratory flow; PEF) in mild asthmatics in a recently published clinical trial of 12 weeks duration (Bateman et al., Ann. Allergy Asthma Immunol., 96(5): 679-86 (2006)). A separate study with roflumilast also demonstrated improvements in airway hyper-responsiveness (AHR) to direct histamine provocation in a similar group of mild asthmatics in response to allergen challenge (Louw et al., Respiration, Sep. 5 2006). Recently published results of a long term (6 month) study of cilomilast treatment in patients with COPD indicated that treatment with a selective PDE4 inhibitor arrested airway function (FEV1) decline in these patients and positively affected their quality of life as measured by the St. Georges Respiratory Questionnaire (Rennard et al., Chest, 129(1) 65-66 (2006)).
The clinical usefulness of PDE4 inhibition has also been demonstrated in disorders of the central nervous system. PDE4 inhibition by rolipram improves cognitive function in rodents and was developed as an antidepressant in humans. cAMP acts as a second messenger for neurotransmitters, and thus mediates their cellular responses. The therapeutic effects of PDE4 inhibitors in cognition and depression likely originate from enhancement of the cAMP-dependent cellular responses.
The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.
International patent application WO 2007/081570 discloses various compounds that may be useful in the treatment of cholesterol-related diseases. However, there is no disclosure of piperidin-2-ones that are substituted at the 3-position with a benzyl group, nor is there any disclosure that such compounds may be useful as phosphodiesterase 4 inhibitors, and therefore in the treatment of inflammation.
International patent application WO 2006/124874 discloses a broad range of inter alia heterocyclic compounds that may be of use as inhibitors of B-Raf, and therefore of use in the treatment of cancer. There is no specific mention in that document of piperidin-2-ones substituted with a phenyl group at the 5-position.
U.S. Pat. No. 6,162,927, US 2002/0055457 and U.S. Pat. No. 7,208,517 and international patent applications WO 2002/11713, WO 2002/011713, WO-99/006397, WO 96/006095, WO 97/030045 and WO 02/017912 all disclose various compounds that may be useful as endothelin antagonists, and therefore of use in the treatment of cancer. There is no specific disclosure in any of these documents of piperidin-2-ones that are substituted at the 5-position with a phenyl group.
US 2007/0203124 discloses various piperazines that may be useful as inhibitors of phosphodiesterase 4 function. However, there is no disclosure in that document of piperidinones.
International patent application WO 2005/115389 discloses various compounds that may be useful in the treatment of negative energy balance in ruminants. However, there is no mention that the compounds disclosed therein may be useful in the treatment of inflammation.
International patent application WO 95/028926 discloses various heterocycles including pyrrolidinones and piperidinones as potentially useful phosphodiesterase 4 inhibitors. However, there is no disclosure in that document of piperidin-2-ones substituted in the 3-position with a benzyl group.
International patent application WO 01/68600 discloses various compounds, including pyrrolidinones, that may be useful in the treatment of inflammation-based diseases. However, there is no disclosure in this document of compounds containing a core piperidin-2-one ring.
Further, US 2003/0186943 and international patent applications WO 00/14083 and WO 2004/031149 disclose inter alia piperidin-2-ones that may be useful in the treatment of inflammation-based diseases. International patent applications WO 2007/137181, WO 2004/091609 and WO 2004/016227 and US application US 2004/0224316 disclose various piperidinones, which may be useful as phosphodiesterase 4 inhibitors. However, only certain 3-benzyl-5-phenylpiperidin-2-ones are disclosed in that document.
Also, European patent EP 299 549 discloses various piperidine derivatives which may have opiate-antagonistic activity. However, there is no mention that such compounds may be useful as phosphodiesterase 4 inhibitors, and therefore of use in the treatment of inflammation.
According to the invention, there is now provided a compound of formula (I),
wherein:
m and q independently represent 0, 1, 2, 3, 4 or 5;
n represents 0, 1, 2 or 3;
r represents 1, 2, 3, 4, 5 or 6;
each R1 independently represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X1), halo, -A1-B1, —R12—CN, —R12—NO2, —R12—N(R10)R11, —R12—OR10, —R12—OC(O)R10, —R12—C(O)R13, —R12—C(O)OR10, —R12—C(O)N(R10)R11, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10, —R12—OS(O)2R10, —R12—S(O)txN(R10)R11, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11, —R12—B(OR10)2, —R12—P(R10)R11, —R12—P(O)(OR10)2 or —R12—OP(O)(OR10)2;
tx represents, on each occasion when used herein, 1 or 2;
px represents, on each occasion when used herein, 0, 1, 2 or 3;
R2 represents hydrogen, —OR4, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X2) or -A2-B2;
R3 represents hydrogen, —OR4, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X2) or -A3-B3;
each R4 independently represents, on each occasion when used herein, hydrogen, —R9—OR10, —R9—C(O)OR10, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X3) and/or -A4-B4;
R5 represents hydrogen, -A5-B5, —R12—C(O)R10, —R12—C(O)OR10, —R12—C(O)N(R10)R11, C1-12 alkyl, C2-12 alkenyl or C2-12 alkynyl, which latter three groups are optionally substituted by one or more substituents selected from X4;
each R6 independently represents halo, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11, —R12—C(O)N(R10)R11, —R12—N(Rw3)C(O)R10, —R12—N(Rw3)C(O)N(R10)R11, —R12—N(Rw3)S(O)tR10x, —R12—N(Rw3)S(O)tOR10x, —R12—OC(O)R10, —R12—OC(O)N(R10)R11, —R12—OS(O)tR10x, —R12—S(O)pR10, —R12—S(O)tN(Rw3)R10, —R12—S(O)tOR10; —R12—Si(R16)3, C1-12 alkyl, C1-12 alkenyl, C1-12 alkynyl, C3-15 cycloalkyl and/or heterocyclyl which latter five groups are optionally substituted by one or more substituents selected from X5; or
any two R6 groups, or R2 and any R6 group, may be linked together to form a further ring, which is formed either by the two relevant groups being linked together by a direct bond or C1-5 alkylene;
each R7 independently represents halo, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11, —R12—C(O)N(R10)R11, —R12—N(Rw3)C(O)R10, —R12—N(Rw3)C(O)N(R10)R11, —R12—N(Rw3)S(O)tR10x, —R12—N(Rw3)S(O)tOR10x, —R12—OC(O)R10, —R12—OC(O)N(R10)R11, —R12—OS(O)tR10x, —R12—S(O)pR10, —R12—S(O)tN(Rw3)R10, —R12—S(O)tOR10; —R12—Si(R16)3, C1-12 alkyl, C1-12 alkenyl, C1-12 alkynyl, C3-15 cycloalkyl and/or heterocyclyl which latter five groups are optionally substituted by one or more substituents selected from X6;
each R8 independently represents hydrogen, —R12—O—R10, -A6-B6, C1-12 alkyl, C2-12 alkenyl or C2-12 alkynyl, which latter three groups are optionally substituted by one or more substituents selected from X7;
each R10x independently represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X8), -A7-O-A8 and/or -A9-B9;
each Rw3, R10 and R11 independently represent, on each occasion when used herein, hydrogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X8), -A7-O-A8 and/or -A9-B9;
or R10 and R11, together with the nitrogen atom to which they are both attached, may be linked together to form a heterocyclyl group (optionally substituted by one or more substituents selected from Z2a) or a heteroaryl group (optionally substituted by one or more substituents selected from Z1a); or
in the case of —R12—B(OR10)2, the two R10 groups may be linked together to form, along with the relevant boron and oxygen atoms, a heterocyclyl group;
each R12 independently represents, on each occasion when used herein, a direct bond or R9;
R13 represents hydrogen, halo, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X9), -A10-O-A11 or -A12-B12; and
each R9 independently represents, on each occasion when used herein, C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene, all of which are optionally substituted by one or more substituents selected from X10;
A1, A3, A4, A9 and A12 independently represent a direct bond, C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene, which latter three groups are optionally substituted by one or more substituents selected from X11;
A2, A5, A6, A7 and A10 independently represent C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene, all of which are optionally substituted by one or more substituents selected from X12;
A8 and A11 independently represent C1-12 alkyl, C2-12 alkenyl or C2-12 alkynyl, all of which are optionally substituted by one or more substituents selected from X13;
B1, B3, B4, B9 and B12 independently represent aryl (optionally substituted by one or more substituents selected from Y1), heteroaryl (optionally substituted by one or more substituents selected from Z1), heterocyclyl (optionally substituted by one or more substituents selected from Z2) or cycloalkyl (optionally substituted by one or more substituents selected from Z3);
B2, B5 and B6 independently represent aryl optionally substituted by one or more substituents selected from Y2;
X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12 and X13 independently represent, on each occasion when used herein, G1, aryl (optionally substituted by one or more T1 substituents), C3-15 cycloalkyl (optionally substituted by one or more T2 substituents), heterocyclyl (optionally substituted by one or more T3 substituents), heteroaryl (optionally substituted by one or more T4 substituents), ═O, —Si(R16)3, —OR14, —OC(O)—R14, —N(R14)2, —C(O)R14, —C(O)OR14, —C(O)N(R14)2, —N(R14)C(O)OR16, —N(R14)C(O)R16, —N(R14)S(O)tR16, —S(O)tOR16, —S(O)pR16, —S(O)tN(R14)2, —N(R14)C(O)N(R14)2, —N(R14)S(O)tOR16, —OC(O)N(R14)2 and/or —OS(O)tR9x;
Y1 and Y2 independently represent, on each occasion when used herein, -Ax-By, G1, G2, —R15—OR17—N(R14)2 and/or —R15—O—R17—N(R14)S(O)tR16;
Z1a, Z1, Z2a, Z2 and Z3 independently represent, on each occasion when used herein, G1, ═O, ═S, -Ax-By and/or G2;
G1 represents C1-12 alkyl (optionally substituted by one or more substituents selected from T5) or C2-12 alkenyl (optionally substituted by one or more substituents selected from T6), halo, —CN, —NO2 or ═O;
G2 represents -Ax-Bx, —R15—OR14, —R15—OC(O)—R14, —R15—N(R14)2, —R15—C(O)R14, —R15—C(O)OR14, —R15—C(O)N(R14)2, —R15—N(R14)C(O)OR16, —R15—N(R14)C(O)R16, —R15—N(R14)S(O)tR16, —R15—S(O)tOR16, —R5—S(O)pR16 and/or —R15—S(O)tN(R14)2;
Ax represents, on each occasion when used herein, a direct bond or C1-12 alkylene optionally substituted by one or more halo or ═O substituents;
Bx represents aryl or heteroaryl, which groups are optionally substituted by one or more substituents selected from T7 and T8, respectively;
By represents cycloalkyl or heterocyclyl, both of which are optionally substituted by one or more substituents selected from halo, C1-6 alkyl (optionally substituted by one or more halo substituents), —OCH3, —OCHF2, —OCF3 and/or ═O;
T1, T4, T5, T6, T7 and T8 independently represent halo, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from Qx1), —OH, —O—C1-6 alkyl, —OC2-6 alkenyl, —OC2-6 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from Qx2), —N(Rw)2, —NO2 and/or —CN; and/or
T5 and T6 may alternatively or additionally represent ═O;
T2 and T3 independently represent halo, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl (which latter three groups are optionally substituted by halo), —OCH3, —OCHF2, —OCF3 and/or ═O;
Qx1 and Qx2 independently represent halo, —OCH3, —OCHF2, —OCF3, —N(Rw)2 and/or ═O;
each Rw independently represents, on each occasion when used herein, hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, which latter three groups are optionally substituted by one or more substituents selected from halo, —OCH3, —OCHF2, —OCF3 and/or ═O; or
two Rw groups, when attached to the same nitrogen atom, may be linked together to form, together with the nitrogen atom to which they are necessarily attached, a 5- or 6-membered ring, optionally containing a further heteroatom and optionally substituted by one or more substituents selected from fluoro, —CH3 and ═O;
t represents, on each occasion when used herein, 1 to 2;
p represents, on each occasion when used herein, 0, 1 or 2;
each R14 independently represents, on each occasion when used herein, hydrogen, -Ax1-Bx1, C1-12 alkyl, C2-6 alkenyl or C2-6 alkynyl, which latter three groups are optionally substituted by one or more substituents selected from E1;
each R15 independently represents, on each occasion when used herein, a direct bond, C1-12 alkylene or C2-12 alkenylene, which latter two groups are optionally substituted by one or more substituents selected from halo, —OCH3, —OCHF2, —OCF3 and ═O;
each R16 independently represents, on each occasion when used herein, C1-12 alkyl, C2-6 alkenyl, C2-6 alkynyl (which latter three groups are optionally substituted by one or more halo and/or ═O groups) or -Ay1-By1;
R17 represents, on each occasion when used herein, C1-12 alkylene or C2-12 alkenylene, both of which are optionally substituted by one or more substituents selected from halo and ═O;
Ax1 and Ay1 independently represent a direct bond or C1-12 (e.g. C1-6) alkylene optionally substituted by one or more halo and/or ═O groups;
Bx1 and By1 independently represent cycloalkyl (e.g. C3-15 cycloalkyl), heterocyclyl (which latter two groups are optionally substituted by one or more substituents selected from halo and ═O), aryl or heteroaryl (which latter two groups are optionally substituted by one or more halo atoms);
E1 represents halo, —CN, —NO2, ═O, —OR18, —OC(O)—R18, —N(R18)2, —C(O)R18, —C(O)OR18, —C(O)N(R18)2, —N(R18)C(O)OR19, —N(R18)C(O)R19, —N(R18)S(O)t1R19, —S(O)t1OR19, —S(O)p1R19, —S(O)t1N(R18)2, —N(R18)C(O)N(R18)2, —N(R18)S(O)t1OR19x, —OC(O)N(R18)2, —OS(O)t1R19x and/or —Si(R19x)3;
each R18 and R19 independently represents, on each occasion when used herein, hydrogen, C1-3 alkyl, C2-3 alkenyl or C2-3 alkynyl, which latter three groups are optionally substituted by one or more halo atoms;
each R19x independently represents, on each occasion when used herein, C1-3 alkyl, C2-3 alkenyl or C2-3 alkynyl, which latter three groups are optionally substituted by one or more halo atoms;
t1 represents, on each occasion when used herein, 1 or 2;
p represents 0, 1 or 2,
or a pharmaceutically acceptable salt, solvate, prodrug or polymorph thereof,
provided that:
(A) when r represents 1, each R8 represents hydrogen, R2 represents hydrogen, m and n both represent 0, R4 represents methyl:
(I) when R3 represents —OR4 in which R4 represents cyclopentyl:
(III) when R3 represents —OR4 in which R4 represents isopropyl:
The skilled person will appreciate that in certain preferred embodiments of the compounds of the invention, some or all of the above provisos will become redundant. For example when “r represents 2, 3, 4, 5 or 6”, then all of the above provisos become redundant. Further, when certain preferred values of R1 and R4 are included (e.g. when “R1 represents —R12—OR9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11” or when “R3 represents —R12—OR9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11”, then, again, all of the above provisos become redundant.
Certain chemical groups named herein are preceded by a shorthand notation indicating the total number of carbon atoms that are to be found in the indicated chemical group. For example, C7-12 alkyl describes an alkyl group, as defined herein, having a total of 7 to 12 carbon atoms, and C4-12 cycloalkylalkyl describes a cycloalkylalkyl group, as defined herein, having a total of 4 to 12 carbon atoms.
The total number of carbons in the shorthand notation does not include carbons that may exist in substituents of the group described.
In addition to the foregoing, as used in the specification and appended claims, unless specified to the contrary, the following terms have the meaning indicated:
“Amino” refers to the —NH2 radical;
“Cyano” refers to the —CN radical;
“Hydroxyl” refers to the —OH radical;
“Imino” refers to the ═NH substituent;
“Nitro” refers to the —NO2 radical;
“Oxo” refers to the ═O substituent;
“Thioxo” refers to the ═S substituent;
“Trifluoromethyl” refers to the —CF3 radical.
Further, “alkyl” refers to cycloalkyl (where there is a minimum of three carbon atoms) or, preferably, a straight or branched hydrocarbon chain radical consisting of carbon and hydrogen atoms, containing no unsaturation. C1-12 alkyl refers to such alkyl groups having from one to twelve carbon atoms, preferably one to eight carbon atoms and, more preferably, one to six carbon atoms, and which group is attached to the rest of the molecule by a single bond. Examples of alkyl groups include methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, and the like.
The term “alkenyl” refers to cycloalkyl containing at least one double bond, or, preferably, refers to a straight or branched hydrocarbon chain radical group consisting of carbon and hydrogen atoms, containing at least one double bond. C2-12 alkenyl refers to such alkenyl groups having from two to twelve carbon atoms, preferably one to eight carbon atoms and, more preferably, one to six carbon atoms, and which group is attached to the rest of the molecule by a single bond. Examples of alkenyl groups include ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like.
The term “alkynyl” refers to cycloalkyl containing at least one triple bond, or, preferably, refers to a straight or branched hydrocarbon chain radical group consisting of carbon and hydrogen atoms, containing at least one triple bond and optionally one or more double bonds. C2-12 alkynyl refers to such alkynyl groups having from two to twelve carbon atoms, preferably one to eight carbon atoms and, more preferably, one to six carbon atoms, and group which is attached to the rest of the molecule by a single bond. Examples of alkynyl groups include ethynyl, prop-1-ynyl, but-1-ynyl, pent-1-ynyl, penta-1-en-4-ynyl, and the like.
The term “alkoxy” when used herein refers to a —O—C1-12 alkyl, in which the C1-12 alkyl group is as defined above (e.g. see the definition of C1-12 alkyl when employed in respect of R1). For example, the relevant C1-12 alkyl group represents C1-12 alkyl optionally substituted by one or more substituents selected from X1.
The term “alkylene” or “alkylene chain” refers to cycloalkylene (when there is a minimum of three carbon atoms) or, preferably, a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting of carbon and hydrogen, and containing no unsaturation. C1-12 alkylene refers to such alkylene groups having from one to twelve carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbon atoms within the chain.
The term “alkenylene” or “alkenylene chain” refers to a cycloalkylene group containing at least one double bond, or, preferably, refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting of carbon and hydrogen, containing at least one double bond. C2-12 alkenylene refers to such alkenylene groups having from two to twelve carbon atoms, e.g., ethenylene, propenylene, n-butenylene, and the like. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain.
The term “alkynylene” or “alkynylene chain” refers to a cycloalkylene group containing at least one triple bond, or, preferably, refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting of carbon and hydrogen, containing at least one triple bond. C2-12 alkynylene refers to such alkynylene groups having from two to twelve carbon atoms, e.g., propynylene, n-butenylene, and the like. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain.
When alkyl, alkenyl, alkynyl, alkylene, alkenylene or alkynylene groups are substituted by a cyclic group, then the point of attachment of the cyclic substituent may be via a single carbon atom.
Unless otherwise specified, such alkyl, alkenyl, alkynyl, alkylene, alkenylene and alkynylene groups that are mentioned (e.g. in the definition of R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12 and/or R13) may be optionally substituted by one or more (e.g. one) X group, i.e. X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12 or X13 (as appropriate).
“Aryl” refers to a hydrocarbon ring system radical comprising from six to eighteen carbon atoms and at least one aromatic ring. For purposes of this invention, the aryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may included fused or bridged ring systems. Aryl radicals include, but are not limited to aryl radicals derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Particular aryl groups that may be mentioned include benzene, naphthene and the like, such as 1,2,3,4-tetrahydronaphthene, indane, indene and fluorene.
Unless otherwise specified, aryl groups that are mentioned (e.g. in the definitions of R1, R2, R3, R5, R8, R10, R11 or R13) may be optionally substituted by one or more (e.g. one) Y group (i.e. Y1 or Y2).
The term “cycloalkyl” refers to a (e.g. stable) non-aromatic monocyclic or polycyclic hydrocarbon radical consisting of carbon and hydrogen atoms, which may include fused or bridged ring systems. C3-15 cycloalkyl refers to such cycloalkyl groups having from three to fifteen carbon atoms, preferably having from three to ten carbon atoms (i.e. C3-10 cycloalkyl), and which group is saturated or unsaturated and attached to the rest of the molecule by a single bond.
Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.
Further cycloalkyl groups that may be mentioned include C3-7 (e.g. C3-6) cycloalkyl groups.
When cycloalkyl, or other cyclic, groups are further substituted by a further cyclic group, then the point of attachment of the cyclic substituent may be via a single carbon atom, so forming a spiro-cyclic compound.
Unless otherwise specified, cycloalkyl groups that are mentioned (e.g. in the definitions of R1, R3, R4, R10, R11 or R13) may be optionally substituted by one or more (e.g. one) Z3 group.
“Halo” refers to halogen and preferably, bromo, chloro, fluoro or iodo.
When used herein, the terms C1-12 haloalkyl, C2-12 haloalkenyl and C2-12 haloalkynyl refer to C1-12 alkyl, C2-12 alkenyl or C2-12 alkynyl, respectively, all of which are as defined herein, but which are substituted by one or more halo groups.
Alkyl groups that may be substituted with halo atoms (i.e. “haloalkyl” groups) include, e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl (1,3-difluoro-2-propyl), 3-bromo-2-fluoropropyl, 1-bromomethyl-2-bromoethyl (1,3-dibromo-2-propyl), and the like. “Haloalkenyl” groups include, e.g., 2,2-difluoroethenyl, 3-chloroprop-1-enyl, and the like. “Haloalkynyl” groups include, e.g., 3-chloroprop-1-ynyl, and the like.
The term ‘hydroxyalkyl’ when used herein refers to a C1-12 alkyl group, as defined herein, but which is substituted by one or more hydroxy (i.e. —OH) groups.
The term “heterocyclyl” refers to a (e.g. stable) 3- to 18-membered non-aromatic ring radical, which consists of two to twelve carbon atoms and from one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. The total number of atoms in the ring system may be between three and twelve (e.g. between five and ten). Unless specifically stated otherwise in the specification, the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic (e.g. monocyclic or bicyclic) ring system, which may include fused or bridged ring systems. The nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidised, the nitrogen atom may be optionally quaternized, and the heterocyclyl radical may be partially or fully saturated. The heterocyclyl group may therefore contain one or more double and/or triple bonds. Heterocyclyl groups that may be mentioned include, but are not limited to 7-azabicyclo-[2.2.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.2.1]-octanyl, 8-azabicyclo[3.2.1]octanyl, aziridinyl, azetidinyl, dihydropyranyl, dihydropyridyl, dihydropyrrolyl (including 2,5-dihydropyrrolyl), dioxolanyl (including 1,3-dioxolanyl), dioxanyl (including 1,3-dioxanyl and 1,4-dioxanyl), dithianyl (including 1,4-dithianyl), dithiolanyl (including 1,3-dithiolanyl), imidazolidinyl, imidazolinyl, morpholinyl, 7-oxabicyclo[2.2.1]heptanyl, 6-oxabicyclo[3.2.1]-octanyl, oxetanyl, oxiranyl, piperazinyl, piperidinyl, pyranyl, pyrazolidinyl, pyrrolidinonyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, sulfolanyl, 3-sulfolenyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydropyridyl (such as 1,2,3,4-tetrahydropyridyl and 1,2,3,6-tetrahydropyridyl), thietanyl, thiiranyl, thiolanyl, thiomorpholinyl, trithianyl (including 1,3,5-trithianyl), tropanyl and the like. Preferred examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Optional substituents on such groups may be attached on any atom. Preferably, the heterocyclyl group is attached to the moiety of formula I via a heterocyclyl ring (i.e a non aromatic ring containing at least one heteroatom) of the polycyclic ring system.
When heterocyclyl groups are further substituted by a cyclic group, then the point of attachment of the cyclic substituent may be via a single atom, so forming a spiro-cyclic compound.
Unless otherwise specified, heterocyclyl groups that are mentioned (e.g. in the definitions of R1, R10, R11 or R13) may be optionally substituted by one or more (e.g. one) Z2 group (eg. Z2 or Z2a).
The term “heteroaryl” refers to a 5- to 18-membered partially or fully aromatic ring radical (i.e. when the heteroaryl group is polycyclic, then at least one of the rings is aromatic), which consists of one to seventeen carbon atoms and from one to ten heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Heteroaryl groups may have between five and fourteen (e.g. between five and ten) members, in which at least one (e.g. one to four) of the atoms in the ring system is/are (a) heteroatom. For purposes of this invention, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic (e.g. monocyclic, bicyclic or tricyclic) ring system, which may include fused or bridged ring systems. However, when the heteroaryl radical is polycyclic (i.e. bicyclic, tricyclic or tetracyclic), then the point of attachment of the heteroaryl group to the other relevant moiety of the compound of formula I is preferably via a heterocyclyl group (i.e. a non-aromatic ring containing at least one heteroatom) or, more preferably a heteroaromatic ring (i.e. an aromatic ring containing at least one heteroatom) of the polycycle. One or more nitrogen, carbon or sulfur atoms (e.g. nitrogen atoms) in the heteroaryl radical may be optionally oxidized, and the nitrogen atom may be optionally quaternized (provided that, when the heteroaryl ring is polycyclic, then the point of attachment with the rest of the compound of formula I is preferably via a ring that remains heteroaromatic). Examples of such groups include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzthiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphtho-furanyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e. thienyl). Particular examples of heteroaryl groups that may be mentioned include benzodioxanyl, benzodioxepinyl, benzodioxolyl (including 1,3-benzodioxolyl), benzofurazanyl, benzoxadiazolyl (including 2,1,3-benzoxadiazolyl), benzoxazinyl (including 3,4-dihydro-2H-1,4-benzoxazinyl), benzomorpholinyl, benzoselenadiazolyl (including 2,1,3-benzoselenadiazolyl), chromanyl, imidazopyridyl (e.g. imidazo[1,2-a]pyridyl), indolinyl, isobenzofuranyl, isochromanyl, isoindolinyl, isoindolyl, isothiochromanyl, phenazinyl, phenothiazinyl, quinolizinyl, quinoxalinyl, thiochromanyl, thiazolopyridyl, tetrahydroisoquinolinyl (including 1,2,3,4-tetrahydroisoquinolinyl and 5,6,7,8-tetrahydroisoquinolinyl), tetrahydroquinolinyl (including 1,2,3,4-tetrahydroquinolinyl and 5,6,7,8-tetrahydroquinolinyl), or, preferably, acridinyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzothienyl (i.e. benzothiophenyl), benzotriazolyl, benzoxazolyl, carbazolyl, cinnolinyl, furanyl, imidazolyl, indazolyl, indolyl, indolizinyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl (including 1,6-naphthyridinyl, 1,5-naphthyridinyl and 1,8-naphthyridinyl), oxadiazolyl (including 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl and, preferably, 1,3,4-oxadiazolyl), oxazolopyridinyl (e.g. oxazolo[5,4-b]pyridinyl, oxazolo[5,4-c]pyridinyl, oxazolo[4,5-b]pyridinyl, oxazolo[4,5-c]pyridinyl), oxazolyl, phthalazinyl, pteridinyl, purinyl, pyrazinyl, pyrazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, thiadiazolyl (including 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl and, preferably, 1,3,4-thiadiazolyl), thiazolyl, tetrazolyl, thiophenyl (i.e. thienyl), triazinyl, and triazolyl (including 1,2,3-triazolyl and 1,2,4-triazolyl).
Unless otherwise specified, heteroaryl groups that are mentioned (e.g. in the definitions of R1, R10, R11 or R13) may be optionally substituted by one or more (e.g. one) Z1 group (e.g. Z1 or Z1a).
As stated above m, when used in respect of the term “—(R6)m”, may represent 0, 1, 2, 3, 4 or 5. For the avoidance of doubt, this means that the piperidin-2-one ring of the compound of formula I may contain no further R6 substituents (when m represent 0), or, may contain up to five R6 substituents at any of the carbon atoms of the piperidin-2-one ring one on which a substituent is not currently specified (i.e. on any carbon atom that is presently substituted with only a hydrogen atom). Similar logic applies to the terms “—(R7)n” and “—(R1)q”, which mean that there are three or five optional substituents present at the free positions of the relevant respective phenyl rings.
It is stated above that any two (of the five possible) R6 groups on the requisite piperidinone ring of the compound of formula I may be linked together to form a further ring, and such groups may be linked together by a direct bond or a C1-5 alkylene linker group. The skilled person will appreciate that when the two relevant R6 groups are on the same or adjacent carbon atoms, then they cannot be linked together by a direct bond to form a further ring (rather, they may only be linked by the C1-5 alkylene group). The two relevant R6 groups may be located on the same carbon atom of the piperidinone ring, in which case they may be linked to form a spiro-cyclic compound. The two relevant R6 groups may also be located on adjacent carbon atoms of the piperidinone ring, so forming a non-bridged fused bicyclic system. Alternatively, the two relevant R6 groups may be located on non-adjacent carbon atoms (and also not on the same carbon atom), so forming a bridged bicyclic ring structure. Similar rings may be formed between R2 and adjacent or non-adjacent R6 groups.
For the avoidance of doubt, in cases in which the identity of two or more substituents in a compound of formula I may be the same, the actual identities of the respective substituents are not in any way interdependent. For example, in the situation in which there are two X1 (or two Z1a) substituents present, then the respective X1 (or Z1a) groups in question may be the same or different. Further, in compounds of formula I, the integer R4 is necessarily present, as is the integer R3, in which R3 may represent —OR4. In these instances, the identities of each R4 substituent is also not to be regarded as being interdependent. Similar logic also applies to the definitions of e.g. R10, R11, etc. Further still, when for example X1 and X4 are each substituted with a G1 group, then the identities of that G1 group is also not to be regarded as being interdependent, i.e. the two G1 moieties may be the same or different, and when both G1 moieties represent C1-12 alkyl substituted by T5, then the T5 integers in each moiety may also be the same or different.
It may be stated herein that various groups are optionally substituted. The skilled person will appreciate that substituents may only be present at a particular position if the rules of valency are adhered to. For example, where it is stated herein that a heteroaryl group may be substituted with an oxo or thioxo group, then the skilled person will appreciate that this is not possible on a carbon atom of the ring system in which the carbon atom is already attached to a double (and a single) bond.
“Prodrugs” is meant to indicate a compound that may be converted under physiological conditions or by solvolysis to a biologically active compound of the invention. Thus, the term “prodrug” refers to a metabolic precursor of a compound of the invention that is pharmaceutically acceptable. A prodrug may be inactive when administered to a subject in need thereof, but is converted in vivo to an active compound of the invention. By the term “prodrug”, we therefore include compounds that form a compound of the invention, in an experimentally-detectable amount, within a predetermined time (e.g. about 1 hour), following oral or parenteral administration. Prodrugs are typically rapidly transformed in vivo to yield the parent compound of the invention, for example, by hydrolysis in blood. The prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in a mammalian organism (see, Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). A discussion of prodrugs is provided in Higuchi, T., et al., “Pro-drugs as Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, Ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated in full by reference herein.
The term “prodrug” is also meant to include any covalently bonded carriers, which release the active compound of the invention in vivo when such prodrug is administered to a mammalian subject. Prodrugs of a compound of the invention may be prepared by modifying functional groups present in the compound of the invention in such a way that the modifications are ‘cleaved’ (i.e. the modified functional group reverts to the original functional group) for example in vivo (i.e. it may be metabolised in the body), to the parent compound of the invention. Prodrugs include compounds of the invention wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the compound of the invention is administered to a mammalian subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol or amide derivatives of amine functional groups in the compounds of the invention and the like.
The invention disclosed herein is also meant to encompass all pharmaceutically acceptable compounds of the invention being isotopically-labelled by having one or more atoms replaced by an atom having a different atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as 2H, 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 31P, 32P, 35S, 18F, 36Cl, 123I and 125I, respectively. These radiolabelled compounds could be useful to help determine or measure the effectiveness of the compounds. Certain isotopically-labelled compounds of the invention, for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, i.e. 3H, and carbon-14, i.e. 14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium, i.e. 2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as 11C, 18F, 15O and 13N, can be useful in Positron Emission Topography (PET) studies. Isotopically-labelled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Examples and Preparations as set out below using an appropriate isotopically-labelled reagent in place of the non-labelled reagent previously employed.
The invention disclosed herein is also meant to encompass the in vivo metabolic products of the disclosed compounds. Such products may result from, for example, the oxidation, reduction, hydrolysis, amidation, esterification, and the like of the administered compound, primarily due to enzymatic processes. Accordingly, the invention includes compounds produced by a process comprising contacting a compound of this invention with a mammal for a period of time sufficient to yield a metabolic product thereof. Such products are typically identified by administering a radiolabelled compound of the invention in a detectable dose to an animal, such as rat, mouse, guinea pig, monkey, or to human, allowing sufficient time for metabolism to occur, and isolating its conversion products from the urine, blood or other biological samples.
“Stable compound” and “stable structure” are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
“Mammal” includes humans and both domestic animals such as laboratory animals and household pets, (e.g. cats, dogs, swine, cattle, sheep, goats, horses, rabbits), and non-domestic animals such as wildlife and the like.
“Optional” or “optionally” means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. For example, “optionally substituted aryl” means that the aryl radical may or may not be substituted and that the description includes both substituted aryl radicals and aryl radicals having no substitution.
“Pharmaceutically acceptable carrier, diluent or excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier, for example one which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.
“Pharmaceutically acceptable salt” includes both acid and base addition salts.
“Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like.
“Pharmaceutically acceptable base addition salt” refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, 2-dimethylaminoethanol (deanol), 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.
Often crystallisations produce a solvate of the compound of the invention. As used herein, the term “solvate” refers to an aggregate that comprises one or more molecules of a compound of the invention with one or more molecules of solvent. The solvent may be water, in which case the solvate may be a hydrate. Alternatively, the solvent may be an organic solvent. Thus, the compounds of the present invention may exist as a hydrate, including a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compound of the invention may be true solvates, while in other cases, the compound of the invention may merely retain adventitious water or be a mixture of water plus some adventitious solvent.
A “pharmaceutical composition” refers to a formulation of a compound of the invention and a medium generally accepted in the art for the delivery of the biologically active compound to mammals, e.g., humans. Such a medium includes all pharmaceutically acceptable carriers, diluents or excipients therefor.
“Therapeutically effective amount” refers to that amount of a compound of the invention which, when administered to a mammal, preferably a human, is sufficient to effect treatment, as defined below, of a disease or condition of interest in the mammal, preferably a human. The effect may be objective (i.e. measurable by some test or marker) or subjective (i.e. the subject gives an indication of or feels an effect).
The amount of a compound of the invention which constitutes a “therapeutically effective amount” will vary depending on several factors including the compound, the condition and its severity, the manner of administration, and the type of mammal to be treated (e.g. the amount may vary depending on the species, age, weight, sex, renal function, hepatic function and response of the mammal), but can be determined routinely by one of ordinary skill in the art having regard to his own knowledge and to this disclosure.
“Treating” or “treatment” as used herein refers to the therapeutic treatment and/or prophylactic treatment of the disease or condition of interest in a mammal, preferably a human, having the disease or condition of interest. Such terms therefore include:
(i) preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed to the condition but has not yet been diagnosed as having it (e.g. prophylactic treatment); or
(ii) therapeutic treatment, i.e. treatment of the disease itself, (e.g. complete or partial treatment), which includes:
As used herein, the terms “disease” and “condition” may be used interchangeably or may be different in that the particular malady or condition may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognised as a disease but only as an undesirable condition or syndrome, wherein a more-or-less specific set of symptoms have been identified by clinicians.
As used herein, the following abbreviations have the indicated meanings:
As stated above, compounds of the invention may exist as a stereoisomers, enantiomers, tautomers, or mixtures thereof.
The compounds of the invention, or their pharmaceutically acceptable salts may contain one or more asymmetric centres and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids. The present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (−), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallisation. Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centres of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included. In certain instances, the spatial orientation of the substituents of the compounds of the invention are designated herein as α (alpha) or β (beta). For purposes of this disclosure, substituents with the α orientation are considered to be below the plane of the paper and substituents with the β orientation are considered to be above the plane of the paper.
A “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are non-superimposeable mirror images of one another.
A “tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule. The present invention includes tautomers of any said compounds.
Also within the scope of the invention is the use of intermediate compounds of the invention (for instance the use of such compounds in a process for preparing compounds of the invention) and the use of all polymorphs of the aforementioned species and crystal habits thereof.
Compounds of the invention that may be mentioned include those in which:
R5 represents hydrogen, -A5-B5, —R12—C(O)R10, —R12—C(O)OR10, —R12—C(O)N(R10)R11 or C1-12 alkyl optionally substituted by one or more substituents selected from X4;
each R6 independently represents halo, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11, —R12—C(O)N(R10)R11 and/or C1-12 alkyl optionally substituted by one or more substituents selected from X5; or
any two R6 groups, or R2 and any R6 group, may be linked together to form a further ring, which is formed either by the two relevant groups being linked together by a direct bond or C1 alkylene;
each R7 independently represents halo, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11, —R12—C(O)N(R10)R11 and/or C1-12 alkyl optionally substituted by one or more substituents selected from X6;
A8 and A11 independently represent C1-12 alkyl optionally substituted by one or more substituents selected from X13;
X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12 and X13 independently represent, on each occasion when used herein, G1, aryl (optionally substituted by one or more T1 substituents), C3-15 cycloalkyl (optionally substituted by one or more T2 substituents), heterocyclyl (optionally substituted by one or more T3 substituents), heteroaryl (optionally substituted by one or more T4 substituents), ═O, —Si(CH3)3, —OR14, —OC(O)—R14, —N(R14)2, —C(O)R14, —C(O)OR14, —C(O)N(R14)2, —N(R14)C(O)OR16, —N(R14)C(O)R16, —N(R14)S(O)tR16, —S(O)tOR16, —S(O)pR16 and/or —S(O)tN(R14)2;
Y1 and Y2 independently represent, on each occasion when used herein, —R15—O—R17—N(R14)S(O)tR16 or, preferably, G1, G2 and/or —R15—OR17—N(R14)2;
T1, T4, T5, T6, T7 and T8 independently represent halo, C1-6 alkyl (optionally substituted by one or more substituents selected from Qx1), —OH, —O—C1-6 alkyl (optionally substituted by one or more substituents selected from Qx2), —N(Rw)2, —NO2 and/or —CN; and/or
T5 and T6 may alternatively or additionally represent ═O;
T2 and T3 independently represent halo, C1-6 alkyl (optionally substituted by halo), —OCH3, —OCHF2, —OCF3 and/or ═O;
Rw represents, on each occasion when used herein, hydrogen or C1-6 alkyl optionally substituted by one or more substituents selected from halo, —OCH3, —OCHF2, —OCF3 and/or ═O; or
two Rw groups, when attached to the same nitrogen atom, may be linked together to form, together with the nitrogen atom to which they are necessarily attached, a 5- or 6-membered ring, optionally containing a further heteroatom and optionally substituted by one or more substituents selected from fluoro, —CH3 and ═O;
each R14 independently represents, on each occasion when used herein, hydrogen, -Ax1-Bx1 or C1-12 alkyl optionally substituted by one or more substituents selected from E1;
each R16 independently represents, on each occasion when used herein, C1-12 alkyl (optionally substituted by one or more halo and/or ═O groups) or -Ay1-By1;
E1 represents —CN, —NO2, or, preferably, halo, ═O, —OR18, —OC(O)—R18, —N(R18)2, —C(O)R18, —C(O)OR18, —C(O)N(R18)2, —N(R18)C(O)OR19, —N(R18)C(O)R19, —N(R18)S(O)t1R19, —S(O)tOR19, —S(O)p1R19 and/or —S(O)t1N(R18)2;
R18 and R19 independently represent, on each occasion when used herein, hydrogen or C1-3 alkyl optionally substituted by one or more halo atoms;
px represents, on each occasion when used herein, 0, 1 or 3.
Compounds of the invention that may be mentioned include those in which:
R5 represents hydrogen, —C(O)R10 or —C(O)OR10;
when R5 represents C1-12 alkyl optionally substituted by one or more substituents selected from X4, then X4 does not represent aryl;
there is no R6 substituent at the 4-position of the piperidin-2-one (i.e. when m is other than 0).
Further compounds of the invention that may be mentioned (preferably for those in which r represents 1 and q is preferably other than 0, i.e. there is at least one
R1 substituent present) include those in which:
R1 represents -A1-B1, —R12—CN, —R12—NO2, —R12—N(R10)R11, —R12—OC(O)R10, —R12—C(O)R13, —R12—C(O)OR10, —R12—C(O)N(R10)R11, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10, —R12—OS(O)2R10, —R12—S(O)txN(R10)R11, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11, —R12—B(OR10)2, —R12—P(R10)R11, —R12—P(O)(OR10)2 or —R12—OP(O)(OR10)2;
R1 represents -A1-B1, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—OS(O)2R10, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11 or —R12—C(═NOH)N(R10)R11;
R1 represents —R12—O—R9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11; and/or
R3 represents —R12—O—R9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11.
Further compounds of the invention that may be mentioned, preferably for those in which r represents 1 (and q is other than 0; i.e. there is at least one R1 substituent present), include those in which:
when R1 represents -A1-B1, —R12—CN, —R12—NO2, —R12—N(R10)R11, —R12—OC(O)R10, —R12—C(O)R13, —R12—C(O)OR10, —R12—C(O)N(R10)R11, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10, —R12—OS(O)2R10, —R12—S(O)txN(R10)R11, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11, —R12—B(OR10)2, —R12—P(R)R11, —R12—P(O)(OR10)2 or —R12—OP(O)(OR10)2, then R3 is as hereinbefore defined;
when R3 represents —R12—O—R9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11, then R1 is as hereinbefore defined;
R1 and R3 are as hereinbefore defined, provided that either:
R1 represents -A1-B1, —R12—CN, —R12—NO2, —R12—N(R10)R11, —R12—OC(O)R10, —R12—C(O)R13, —R12—C(O)OR10, —R12—C(O)N(R10)R11, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10, —R12—OS(O)2R10, —R12—S(O)txN(R10)R11, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11, —R12—B(OR10)2, —R12—P(R10)R11, —R12—P(O)(OR10)2 or —R12—OP(O)(OR10)2; or
R3 represents —R12—O—R9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11.
Yet further compounds of the invention that may be mentioned, preferably for those in which r represents 1 (and q is other than 0), include those in which:
when R1 represents -A1-B1, —R12—O—R9—C(O)N(R10)R11, —R12—OS(O)2R10, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11 or, preferably, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, then R3 is as hereinbefore defined;
R1 and R3 are as hereinbefore defined, provided that either:
R1 represents -A1-B1, —R12—O—R9—C(O)N(R10)R11, —R12—OS(O)2R10, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11 or, preferably, —R12—N(R10)C(O)N(R10)R11 or —R12—O—R9—C(O)OR10; or
R3 represents —R12—O—R9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11;
for example when r represents 1, then most preferably at least one of R1 and R3 represents —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10.
Further compounds of the invention that may be mentioned include those in which:
r represents 2, 3, 4, 5 or 6;
for example when r represents 1, then at least one (i.e. one or both) R8 groups represent a substituent other than hydrogen (i.e. —R12—O—R10, -A6-B6, C1-12 alkyl, C2-12 alkenyl or C2-12 alkynyl, which latter three groups are optionally substituted by one or more substituents selected from X7);
for example when r represents 1, then R3 represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X2; preferably in which X represents aryl, e.g. phenyl, optionally substituted as hereinbefore defined) or -A3-B3 (e.g. in which A3 represents a direct bond and B3 preferably represent phenyl);
for example when r represents 1, then R5 represents C1-12 (e.g. C1-6) alkyl (optionally substituted by one or more substituents selected from X4) or, preferably, —R12—C(O)R10 or, preferably, —R12—C(O)N(R10)R11;
when r represents 1, then R2 represents a substituent other that hydrogen, e.g. C1-12 alkyl (optionally substituted by one or more substituents selected from X2) or, preferably, -A2-B2 (in which A2 represents a direct bond or, preferably, C1-3 alkylene (e.g. —CH2—) and B2 preferably represents an optionally substituted phenyl group that is preferably the same as the requisite phenyl-(R1)q group of formula I;
when r represents 1 (and q is other than 0), then R12 preferably represents an optionally substituted C1-12 alkylene group (e.g. C1-3 alkylene), for example when R1 represents —R12—CN, —R12—NO2, —R12OR10—R12—OC(O)R10, —R12—C(O)R13, —R12—C(O)OR10, —R12—C(O)N(R10)R11, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10, —R12—OS(O)2R10, —R12—S(O)txN(R10)R11, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11, —R12—B(OR10)2, —R12—P(R10)R11, —R12—P(O)(OR10)2, —R12—OP(O)(OR10)2 or, more preferably, —R12—N(R10)R11.
Preferred compounds of the invention that may be mentioned include those in which:
B3 and B4 independently represent aryl (optionally substituted by one or more substituents selected from Y1) or cycloalkyl (optionally substituted by one or more substituents selected from Z3);
Y1 and Y2 independently represent, on each occasion when used herein, —R15—O—R17—N(R14)S(O)tR16 or, more preferably, G1, G2 and/or —R15—OR17—N(R14)2.
Compounds of the invention that may be mentioned include those in which:
each R1 independently represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C1-12 hydroxyalkyl, halo, C1-12 haloalkyl, C2-12 haloalkenyl, C2-12 haloalkynyl, optionally substituted -A1-B1, —R12—CN, —R12—NO2, —R12—N(R10)R11, —R12—OR10, —R12—OC(O)R10, —R12—C(O)R13, —R12—C(O)OR10, —R12—C(O)N(R10)R11, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10, —R12—OS(O)2R10, —R12—S(O)txN(R10)R11, —R12—N(R10)S(O)tx N(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11, —R12—B(OR10)2, —R12—P(R10)R11, —R12—P(O)(OR10)2 or —R12—OP(O)(OR10)2;
A1 represents a direct bond, C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene;
B1 represents aryl, heteroaryl or heterocyclyl;
R2 represents hydrogen, C1-12 alkyl, C1-12 haloalkyl or optionally substituted -A2-B2;
A2 represents C1-12 alkylene;
B2 represents aryl;
R3 represents hydrogen, —OR4, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl or optionally substituted -A3-B3;
A3 represents a direct bond or C1-12 alkylene;
B3 represents aryl or cycloalkyl;
each R4 is independently selected from the group consisting of hydrogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl and/or optionally substituted -A4-B4;
A4 represents a direct bond, C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene;
B4 represents cycloalkyl or aryl;
when B4 represents cycloalkyl, then A4 represents a direct bond, C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene;
when B4 represents aryl, then A4 preferably represents a direct bond or C1-12 alkylene;
R5 represents hydrogen, C1-12 alkyl, C1-12 haloalkyl, optionally substituted -A5-B5, —R12—C(O)R10, —R12—C(O)OR10 or —R12—C(O)N(R10)R11;
A5 represents C1-12 alkylene;
B5 represents aryl;
each R6 and each R7 is independently selected from the group consisting of C1-12 alkyl, halo, C1-12 haloalkyl, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11 and —R12—C(O)N(R10)R11;
any two R6 groups, or R2 and any R6 group, are preferably not linked together;
each R8 independently represents hydrogen, —R12—O—R10, C1-12 alkyl, C1-12 haloalkyl or optionally substituted -A6-B6;
A6 represents C1-12 alkylene;
B6 represents aryl;
each R10 and R11 is independently selected from the group consisting of hydrogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C1-12 haloalkyl, C1-12 haloalkenyl, -A7-O-A8 (e.g. —C1-12 alkylene-O—C1-12 alkyl) and optionally substituted -A9-B9;
or R10 and R11, together with the nitrogen atom to which they are both attached, may represent an optionally substituted heterocyclyl or an optionally substituted heteroaryl group; or
in the case of —R12—B(OR10)2, two —OR10 groups, together with the boron atom to which they are both attached, may form an optionally substituted heterocyclyl group;
A9 represents a direct bond or C1-12 alkylene;
B9 represents cycloalkyl, aryl, heterocyclyl or heteroaryl;
each R12 independently represents, on each occasion when used herein, a direct bond or R9;
each R9 independently represents straight or branched optionally substituted C1-12 alkylene; straight or branched optionally substituted C2-12 alkenylene; or straight or branched optionally substituted C2-12 alkynylene; and/or
R13 represents hydrogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, halo, C1-12 haloalkyl, C1-12 haloalkenyl, -A10-O-A11 (e.g. —C1-12 alkylene-O—C1-12 alkyl) or optionally substituted -A12-B12;
A7 and A10 independently represent C1-12 alkylene;
A12 represents a direct bond or C1-12 alkylene;
B12 represents cycloalkyl, aryl, heterocyclyl or heteroaryl;
X1 to X13 independently represent C1-12 alkyl, C2-12 alkenyl, halo, C2-12 haloalkenyl, —CN, —NO2, aryl, C3-15 cycloalkyl, heterocyclyl, heteroaryl, ═O, —Si(CH3)3, —OR14, —OC(O)—R14, —N(R14)2, —C(O)R14, —C(O)OR14, —C(O)N(R14)2, —N(R14)C(O)OR16, —N(R14)C(O)R16, —N(R14)S(O)tR16, —S(O)tOR16, —S(O)pR16 and/or —S(O)tN(R14)2;
each Z1 and Z1a independently represent C1-12 alkyl, C2-12 alkenyl, C1-12 alkoxy, halo, C2-12 haloalkyl, C2-12 haloalkenyl, —CN, ═O, ═S, —NO2, -Ax-Bx, -Ax-By, —R15—OR14, —R15—OC(O)—R14, —R15—N(R14)2, —R15—C(O)R14, —R15—C(O)OR14, —R15—C(O)N(R14)2, —R15—N(R14)C(O)OR16, —R15—N(R14)C(O)R6, —R15—N(R14)S(O)tR16, —R15—S(O)tOR16, —R15—S(O)pR16 and/or —R15—S(O)tN(R14)2;
each Z2 and Z2a independently represent C1-12 alkyl, C2-12 alkenyl, halo, C2-12 haloalkyl, C2-12 haloalkenyl, —CN, ═O, ═S, —NO2, -Ax-Bx, -Ax-By, —R15—OR14, —R15—OC(O)—R14, —R15—N(R14)2, —R15—C(O)R14, —R15—C(O)OR14, —R15—C(O)N(R14)2, —R15—N(R14)C(O)OR16, —R15—N(R14)C(O)R16, —R15—N(R14)S(O)tR16, —R15—S(O)tOR16, —R15—S(O)R16 and/or —R15—S(O)tN(R14)2;
each Z3 independently represents C1-12 alkyl, C2-12 alkenyl, halo, C2-12 haloalkyl, C2-12 haloalkenyl, —CN, ═O, —NO2, -Ax-Bx, -Ax-By, —R15—OR14, —R15—OC(O)—R14, —R15—N(R14)2, —R15—C(O)R14, —R15—C(O)OR14, —R15—C(O)N(R14)2, —R15—N(R14)C(O)OR16, —R15—N(R14)C(O)R16, —R15—N(R14)S(O)tR16, —R15—S(O)tOR16, —R15—S(O)pR16 and/or —R15—S(O)tN(R14)2;
each Y1 and Y2 independently represent C1-12 alkyl, C2-12 alkenyl, halo, C2-12 haloalkyl, C2-12 haloalkenyl, —CN, —NO2, -Ax-Bx, —R15—OR14, —R15—OC(O)—R14, —R15—N(R14)2, —R15—O—R17—N(R14)2, —R15—C(O)R14, —R15—C(O)OR14, —R15—C(O)N(R14)2, —R15—N(R14)C(O)OR16, —R15—N(R14)C(O)R16, —R15—N(R14)S(O)R16, —R15—S(O)tOR16, —R15—S(O)pR16 and/or —R15S(O)tN(R14)2;
t represents, on each occasion when used herein, 1 or 2;
p represents, on each occasion when used herein, 0, 1 or 2;
Ax represents C1-12 alkylene or, preferably a direct bond;
Bx represents aryl or heteroaryl;
By represents cycloalkyl or heterocyclyl;
when Z1, Z1a, Z2 or Z2a represents a group containing R14, then each R14 independently represents, on each occasion when used herein, hydrogen, C1-12 alkyl, C2-12 alkenyl, C1-12 haloalkyl or -Ax1-Bx1;
when an X group (i.e. X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12 or X13), Z3, Y1 or Y2 represents a group containing R14, then each R14 independently represents, on each occasion when used herein, hydrogen, C1-12 alkyl, C1-12 haloalkyl or -Ax1-Bx1;
when an X group, Z3a, Z3b, Y1, Y2 or Y3 represents a group containing R16, then each R16 independently represents, on each occasion when used herein, C1-12 alkyl, C1-12 haloalkyl or -Ay1-By1;
each R15 independently represents, on each occasion when used herein, a direct bond, C1-12 alkylene or C2-12 alkenylene;
when Z1, Z1a, Z2 or Z2a represents a group containing R16, then each R16 independently represents, on each occasion when used herein, C1-12 alkyl, C2-12 alkenyl, C1-12 haloalkyl or -Ay1-By1;
R17 represents C1-12 alkylene or C2-12 alkenylene; and/or
Ax1 and Ay1 independently represent a direct bond or C1-12 (e.g. C1-6) alkylene;
Bx1 and By1 independently represent C3-15 cycloalkyl, heterocyclyl, aryl or heteroaryl.
Further preferred compounds of the invention include those in which:
R2 represents hydrogen, -A2-B2 or C1-12 alkyl which latter two groups are optionally substituted by one or more substituents selected from X2;
each R6 and R7 independently represents halo, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11, —R12—C(O)N(R10)R11 and/or C1-12 alkyl optionally substituted by one or more substituents selected from X5 or X6 (as appropriate);
each R8 independently represents hydrogen, —R12—O—R10, -A6-B6 or C1-12 alkyl optionally substituted by one or more substituents selected from X7;
A2, A5, A6, A7 and A10 independently represent C1-12 alkylene optionally substituted by one or more substituents selected from X12;
A3, A9 and A12 independently represent a direct bond or C1-12 alkylene optionally substituted by one or more substituents selected from X11;
each R4 independently represents hydrogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X3) and/or -A4-B4;
B1 represents aryl, heteroaryl or heterocyclyl, all of which are optionally substituted by one or more substituents selected from Y1, Z1 or Z2, respectively;
B4 represents heterocyclyl (optionally substituted as defined herein) or, preferably, C3-15 cycloalkyl (optionally substituted as defined herein);
Z3a and Z3b independently represent, on each occasion when used herein, G1, ═O, -Ax-By and/or G2;
T5 and T6 independently represent halo;
G1 represents —NO2 or, more preferably, halo, —CN, C1-12 alkyl (optionally substituted by one or more substituents selected from T5) or C2-12 alkenyl (optionally substituted by one or more substituents selected from T6);
when any of X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12 and X13 represent aryl, C3-15 cycloalkyl, heterocyclyl or heteroaryl, then such groups are optionally substituted as defined herein, or, more preferably unsubstituted;
when G1 represents C2-12 alkenyl, then it may be unsubstituted or substituted by one or more halo atoms;
when G1 represents C1-12 alkyl or C2-12 alkenyl, then such groups may be optionally substituted as defined herein, or, are more preferably, unsubstituted;
when Ax represents C1-12 alkylene, then this group may be optionally substituted as hereinbefore defined, but is preferably unsubstituted;
when Bx represents aryl or heteroaryl, then these groups may be optionally substituted as hereinbefore defined, but are preferably unsubstituted;
when By represents cycloalkyl or heterocyclyl then these groups may be optionally substituted as hereinbefore defined, but are preferably unsubstituted;
when R15 represents C1-12 alkylene or C2-12 alkenylene, then such groups may be optionally substituted as defined herein, or, are preferably unsubstituted;
each R16 independently represents, on each occasion when used herein, -Ay1-By1 or C1-12 alkyl optionally substituted by one or more halo atoms;
when R17 represents C1-12 alkylene or C2-12 alkenylene, then such groups may be optionally substituted as defined herein, or, are preferably unsubstituted;
when Ax1 and Ay1 represent C1-12 (e.g. C1-6) alkylene, then that group may be optionally substituted as defined herein, or, is preferably unsubstituted;
when Bx1 and By1 represent C3-15 cycloalkyl, aryl, heterocyclyl or heteroaryl, then such groups may be optionally substituted as defined herein, or, are preferably unsubstituted;
when R14 represents C1-12 alkyl, then such a group is preferably unsubstituted, and, when it is substituted, then it is preferably substituted by one or more halo atoms;
each R14 independently represents, on each occasion when used herein, hydrogen, -Ax1-Bx1 or C1-12 alkyl, which latter group may be unsubstituted or is substituted by one or more substituents selected from halo;
E1 represents ═O, —OR18, or, more preferably, halo or —C(O)N(R18)2;
R18 and R19 independently represent hydrogen;
when R10 and R11, together with the nitrogen atom to which they are both attached, linked together to form an optionally substituted heterocyclyl or an optionally substituted heteroaryl group, then such groups are preferably 5- to 10-membered monocyclic or bicyclic groups, preferably containing one to three (e.g. one or two) heteroatoms selected from sulfur or, preferably nitrogen or oxygen.
Preferred compounds of the invention (e.g. when r represents 1) include those in which:
m represents 1, 2, 3, 4, 5 or, preferably, 0;
q represents 3, 4, 5, preferably, 0, 2 or, more preferably 1;
n represents 1, 2, 3 or, preferably, 0;
r represents 1;
each R1 independently represents optionally substituted -A1-B1, preferably, —R12—N(R10)C(O)N(R10)R11, —R12—OS(O)2R10, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11 or, more preferably, —R12—O—R9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11; or
R1 may also represent C2-12 alkenyl, C2-12 alkynyl or, preferably, C1-12 alkyl when
R3 represents —R12—O—R9—C(O)OR10 or —R12—O—R9—C(O)N(R10)R11;
tx represents, on each occasion when used herein, 1 or 2;
A1 represents C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene (e.g. all of which are optionally substituted by one or more substituents selected from X11);
B1 represents heteroaryl or heterocyclyl (e.g. both of which are optionally substituted by one or more substituents selected from Z1 or Z2, respectively);
R2 represents —OR4, preferably, C1-12 haloalkyl, optionally substituted -A2-B2 or, more preferably, hydrogen or C1-12 alkyl (and most preferably R2 represents hydrogen);
A2 represents C1-12 alkylene;
B2 represents aryl;
R3 represents C2-12 alkenyl, C2-12 alkynyl, preferably, hydrogen, C1-12 alkyl, optionally substituted -A3-B3 or, more preferably, —OR4, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11;
A3 represents a direct bond or C1-12 alkylene;
B3 represents cycloalkyl or aryl;
each R4 independently represents, on each occasion when used herein, C2-12 alkenyl, C2-12 alkynyl, preferably, hydrogen, or, more preferably, C1-12 alkyl or optionally substituted -A4-B4;
A4-B4 represents C1-12 alkylene-cycloalkyl, C2-12 alkenylene-cycloalkyl, C2-12 alkynylene-cycloalkyl or, preferably, cycloalkyl, aryl or C1-12 alkylene-aryl;
A4 represents C2-12 alkenylene, C2-12 alkynylene or, preferably, a direct bond or C1-12 alkylene;
when B4 represents cycloalkyl, then A4 represents C1-12 alkylene, C2-12 alkenylene, C2-12 alkynylene or, more preferably, a direct bond;
R5 represents C1-12 alkyl, C1-12 haloalkyl, optionally substituted -A5-B5, —R12—C(O)R10, —R12—C(O)OR10, —R12—C(O)N(R10)R11 or, preferably, hydrogen;
A5 represents C1-12 alkylene;
B5 represents aryl;
each R6 and each R7 is independently selected from the group consisting of C1-12 alkyl, halo, C1-12 haloalkyl, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11 and —R12—C(O)N(R10)R11;
each R8 independently represents —R12—OR10, optionally substituted -A6-B6, C1-12 alkyl, C1-12 haloalkyl or, preferably, hydrogen;
A6 represents C1-12 alkylene;
B6 represents aryl;
each R10 and R11 independently represent, on each occasion when used herein, C2-12 alkynyl, preferably, C2-12 alkenyl, C2-12 haloalkenyl or, more preferably, hydrogen, C1-12 alkyl, C1-12 haloalkyl, -A7-O-A8 (e.g. —C1-12 alkylene-O—C1-12 alkyl) and/or optionally substituted -A9-B9; or
R10 and R11, together with the nitrogen atom to which they are both attached, may form optionally substituted heterocyclyl or optionally substituted heteroaryl (although R10 and R11 are preferably not linked together);
A9 represents a direct bond or C1-12 alkylene;
B9 represents cycloalkyl, aryl, heterocyclyl or heteroaryl;
each R12 independently represents, on each occasion when used herein, a direct bond or R9; and/or
each R9 independently represents, on each occasion when used herein, straight or branched optionally substituted C2-12 alkenylene, straight or branched optionally substituted C2-12 alkynylene, or, more preferably, straight or branched optionally substituted C1-12 alkylene.
Further preferred compounds of the invention (e.g. when r represents 2, 3, 4, 5 or 6) include those in which:
m represents 1, 2, 3, 4, 5 or, preferably, 0;
q represents 3, 4, 5 or, preferably, 0, 1 or 2;
n represents 1, 2, 3 or, preferably, 0;
r represents 2, 3, 4, 5 or 6;
each R1 independently represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C1-12 hydroxyalkyl, halo, C1-12 haloalkyl, C2-12 haloalkenyl, C2-12 haloalkynyl, optionally substituted -A1-B1, —R12—CN, —R12—NO2, —R12—N(R10)R11, —R12—OR10, —R12—OC(O)R10, —R12—C(O)R13, —R12—C(O)OR10, —R12—C(O)N(R10)R11, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10, —R12—OS(O)2R10—R12—S(O)txN(R10)R11, —R12—N(R10)S(O)txN(R10)R11, —R12—C(═NR10)N(R10)R11, —R12—C(═NOH)N(R10)R11, —R12—B(OR10)2, —R12—P(R10)R11, —R12—P(O)(OR10)2 or —R12—OP(O)(OR10)2;
px represents, on each occasion when used herein, 0, 1 or 3;
tx represents, on each occasion when used herein, 1 or 2;
A1 represents C1-12 alkylene, C2-12 alkenylene or C2-12 alkynylene (e.g. all of which are optionally substituted by one or more substituents selected from X11);
B1 represents aryl, cycloalkyl, heteroaryl or heterocyclyl (e.g. all of which are optionally substituted by one or more substituents selected from Y1, Z3a, Z1 or Z2, respectively);
R2 represents C1-12 alkyl, C1-12 haloalkyl, optionally substituted -A2-B2 or, more preferably, hydrogen;
A2 represents C1-12 alkylene;
B2 represents aryl;
R3 represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, optionally substituted -A3-B3 or, preferably, hydrogen, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11 or, more preferably, —OR4;
A3 represents a direct bond or C1-12 alkylene;
B3 represents cycloalkyl or aryl;
each R4 independently represents, on each occasion when used herein, C2-12 alkenyl, C2-12 alkynyl, preferably, hydrogen, or, more preferably, C1-12 alkyl or optionally substituted -A4-B4;
A4-B4 represents C1-12 alkylene-cycloalkyl, C2-12 alkenylene-cycloalkyl, C2-12 alkynylene-cycloalkyl or, preferably, aryl or C1-12 alkylene-aryl or, more preferably, cycloalkyl;
A4 represents C2-12 alkenylene, C2-12 alkynylene, preferably, C1-12 alkylene or, more preferably, a direct bond;
when B4 represents cycloalkyl, then A4 represents C1-12 alkylene, C2-12 alkenylene, C2-12 alkynylene or, more preferably, a direct bond;
R5 represents C7-12 alkyl, C1-12 haloalkyl, optionally substituted -A5-B5, —R12—C(O)R10, —R12—C(O)OR10, —R12—C(O)N(R10)R11 or, preferably, hydrogen;
A5 represents C1-12 alkylene;
B5 represents aryl;
each R6 and each R7 is independently selected from the group consisting of C1-12 alkyl, halo, C1-12 haloalkyl, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11 and —R12—C(O)N(R10)R11;
each R8 independently represents —R12—OR10, optionally substituted -A6-B6, C1-12 alkyl, C1-12 haloalkyl or, preferably, hydrogen;
A6 represents C1-12 alkylene;
B6 represents aryl;
each R10 and R11 independently represent, on each occasion when used herein, C2-12 alkenyl, C2-12 alkynyl, C2-12 haloalkenyl, preferably, C1-12 haloalkyl, -A7-O-A8 (e.g. —C1-12 alkylene-O—C1-12 alkyl) or, more preferably, hydrogen, C1-12 alkyl and/or optionally substituted -A9-B9; or
R10 and R11, together with the nitrogen atom to which they are both attached, may form optionally substituted heterocyclyl or optionally substituted heteroaryl (although R10 and R11 are preferably not linked together);
A9 represents a direct bond or C1-12 alkylene;
B9 represents cycloalkyl, heterocyclyl, heteroaryl or, preferably, aryl;
each R12 independently represents, on each occasion when used herein, a direct bond or R9; and/or
each R9 independently represents, on each occasion when used herein, straight or branched optionally substituted C2-12 alkenylene, straight or branched optionally substituted C2-12 alkynylene, or, more preferably, straight or branched optionally substituted C1-12 alkylene.
Yet more preferred compounds of the invention (e.g. when r represents 1) include those in which:
m represents 0;
q represents 0, 1 or 2;
n represents 0;
r represents 1;
each R1 independently represents C1-12 alkyl, halo, C1-12 haloalkyl, —R12—CN, —R12—N(R10)R11 or —R12—OR10;
R2 represents hydrogen or optionally substituted -A2-B2;
A2 represents C1-12 alkylene;
B2 represents aryl optionally substituted with one or more substituents selected from Y2;
Y2 represents G1 or G2;
G1 represents halo or C1-12 haloalkyl;
G2 represents —R15—OR14 or —R15—N(R14)2;
each R14 independently represents hydrogen, C1-12 alkyl, C1-12 haloalkyl or -Ax1-Bx1;
Ax1 represents a direct bond or C1-12 (e.g. C1-6) alkylene;
Bx1 represents cycloalkyl, aryl, heterocyclyl or heteroaryl;
each R15 independently represents a direct bond or straight or branched C1-12 alkylene;
R3 represents —OR4 or optionally substituted -A3-B3;
A3 represents a direct bond;
B3 represents aryl;
each R4 independently represents C1-12 alkyl or optionally substituted -A4-B4;
A4-B4 preferably represents cycloalkyl;
A4 represents a direct bond;
B4 represents cycloalkyl;
R5 represents hydrogen, C1-12 alkyl, -A5-B5, —R12—C(O)R10 or —R12—C(O)N(R10)R11;
A5 represents C1-12 alkylene;
B5 represents aryl;
each R8 independently represents hydrogen, —R12—OR10 or C1-12 alkyl;
each R10 and R11 independently represent, on each occasion when used herein, hydrogen, C1-12 alkyl, C1-12 haloalkyl and/or optionally substituted -A9-B9;
R10 and R11 are preferably not linked together;
A9-B9 represents cycloalkyl, aryl or C1-12 alkylene-aryl;
A9 represents a direct bond or C1-12 alkylene;
B9 represents cycloalkyl or aryl;
when B9 represents aryl, then A9 preferably represents a direct bond or C1-12 alkylene;
when B9 represents cycloalkyl, then A9 preferably represents a direct bond;
each R12 independently represents, on each occasion when used herein, a direct bond or R9; and/or
each R9 independently represents, on each occasion when used herein, straight or branched optionally substituted C1-12 alkylene.
Preferred compounds of the invention include those in which:
m, n and q independently represent 0, 1, or 2;
R2 represents hydrogen, C1-12 (e.g. C1-6) alkyl or C1-12 (e.g. C1-6) alkenyl, which latter two groups are optionally substituted by one or more substituents selected from X2 (such as hydroxy or, preferably, halo);
R3 represents -A3-B3 or, preferably, hydrogen, —OR4, —R12—O—R9—C(O)R10 or —R12—O—R9—C(O)N(R10)R11;
each R4 independently represents hydrogen, —R9—OR10, —R9—C(O)OR10, C1-12 (e.g. C1-6) alkyl (optionally substituted by one or more substituents selected from X3) or -A4-B4
A3 and A4 independently represent C1-3 alkylene or, preferably, a direct bond;
B4 represents C3-15 (e.g. C5-10) cycloalkyl (optionally substituted by one or more substituents selected from Z3) or a 3- to 18- (e.g. 5- to 10-) membered heterocyclyl group (optionally substituted by one or more substituents selected from Z2);
R5 represents -A5-B5 or, preferably, hydrogen, —R12C(O)R10 or —R12—C(O)N(R10)R11;
each R6 and R7 independently represent halo, —R12—OR10, —R12—CN, —R12—NO2, —R12—C(O)OR10, —R12—N(R10)R11, —R12—C(O)N(R10)R11 and/or C1-12 (e.g. C1-6) alkyl optionally substituted by one or more substituents selected from X5 or X6 as appropriate (e.g. halo; so forming a haloalkyl group);
R8 represents hydrogen or C1-6 (e.g. C1-3) alkyl optionally substituted by one or more substituents selected from X7;
each R9 independently represents C1-12 alkylene optionally substituted by one or more substituents selected from X10;
each R10 and R11 independently represent hydrogen, C1-12 (e.g. C1-6) alkyl, C1-12 (e.g. C1-6) alkenyl (which latter two groups are optionally substituted by one or more substituents selected from X8 (e.g. halo)), -A7-O-A8 and/or -A9-B9; or
R10 and R11 are linked together to form, together with the nitrogen atom to which they are necessarily attached, a 5- or 6-membered heterocyclyl group (e.g. pyrrolidinyl, piperidinyl, morpholinyl or piperazinyl), which is optionally substituted by one or more substituent selected from Z2a (e.g. halo, —CH3 and ═O);
R13 represents hydrogen or C1-6 (e.g. C1-3) alkyl optionally substituted by one or more substituents selected from X9;
A1, A3, A4, A9 and A12 independently represent C1-6 (e.g. C1-3) alkylene (optionally substituted as defined herein, e.g. by one or more X11 substituents);
A8 and A11 independently represent C1-6 (e.g. C1-3) alkyl (optionally substituted as defined herein);
A2, A5, A6, A7 and A10 independently represent a direct bond or C1-6 (e.g. C1-3) alkylene (optionally substituted as defined herein, e.g. by one or more X12 substituents);
X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12 and X13 independently represent a 5- or 6-membered heterocyclyl group (preferably containing a nitrogen heteroatom and optionally a further nitrogen or oxygen heteroatom; optionally substituted by one or more T3 substituents), —OR14, N(R14)2, —C(O)OR14, —C(O)N(R14)2, —S(O)tN(R14)2 or more preferably, G1;
t represents 2;
Y1 and Y2 independently represent G2 or preferably, G1;
Z1a, Z1, Z2a, Z2 and Z3 independently represent ═O or, preferably, G2 or G1;
G1 represents —CN, —NO2 or, preferably, halo or C1-6 (e.g. C1-3) alkyl optionally substituted by one or more T5 substituents (such as halo (e.g. fluoro) atoms);
G2 represents —R15—N(R14)2, —R15—C(O)OR14, —R15—C(O)N(R14)2, —R15—S(O)tN(R14)2 or more preferably —R15—O—R14;
T1, T2, T3, T4, T5, T6, T7, T and T8 independently represent halo (e.g. chloro or fluoro) or C1-3 alkyl optionally substituted by one or more Qx1 or halo substituents as appropriate);
Qx1 and Qx2 independently represent halo (e.g. chloro or fluoro);
R15 represents a direct bond;
R14 represents C1-6 (e.g. C1-3) alkyl (e.g. methyl) optionally substituted by one or more substituents selected from E1 (e.g. —C(O)N(R18)2 and halo (e.g. fluoro));
each R16 independently represents C1-3 alkyl (optionally substituted by one or more fluoro atoms);
R17 represents C1-6 (e.g. C1-3) alkylene;
R18 and R19 independently represent hydrogen.
Preferred compounds of the invention include those in which:
R2 represents -A2-B2, C1-4 alkyl or, more preferably, hydrogen;
R3 represents -A3-B3 or, preferably, —R12—O—R9—C(O)R10, —R12—O—R9—C(O)N(R10)R11 or —OR4;
R4 represents -A4-B4 or a C1-6 alkyl group, for instance a straight-chain alkyl (e.g. isopropyl, n-propyl, ethyl or, preferably, methyl) group optionally substituted by one or more substituents selected from X3, for example —OR14, —C(O)OR14 or, preferably, fluoro (so forming, for example, a difluoromethyl or trifluoromethyl group);
X1 represents —OR14 (in which R14 preferably represents hydrogen);
X3 represents —OR14 (in which R14 preferably represents hydrogen), —C(O)OR14 (in which R14 preferably represents hydrogen or C1-2 alkyl) or, preferably, G1;
A3 represents a direct bond;
A4 represents a C1-2 methylene (e.g. —CH2—) or, preferably, direct bond;
B3 represents aryl (e.g. phenyl), which group is optionally substituted by one or more substituents selected from Y1, but is preferably unsubstituted;
B4 represents a C3-5 cycloalkyl (e.g. a C3 cyclopropyl group or, preferably a C5 cyclopentyl group) or a 4- to 6- (e.g. a 5- or 6-) membered heterocyclyl group (e.g. a 4- or, preferably, a 5-membered heterocyclyl group containing one oxygen atom, such as oxetanyl, e.g. 3-oxetanyl, or, more preferably, tetrahydrofuranyl, e.g. 3-tetrahydrofuranyl, group);
R5 represents -A5-B5, —R12—C(O)R10 (in which R12 is preferably a direct bond), —R12—C(O)N(R10)R11 (in which R12 is preferably C1-3 alkaline, such as —CH2—, or, preferably, hydrogen;
each R6 and R7 independently represent halo or C1-12 (e.g. C1-6) alkyl optionally substituted by one or more substituents selected from X5 or X6 as appropriate (e.g. halo; so forming a haloalkyl group);
R8 represents hydrogen or C1-6 (e.g. C1-3) alkyl (e.g. methyl);
A1 and A2 independently represent C1-3 (e.g. C1-2) alkylene, such as ethylene or, preferably, methylene (which group is optionally substituted by one or more halo, e.g. fluoro, atoms, or, preferably unsubstituted);
B1 represents aryl (optionally substituted by one or more substituents selected from Y1) or heteroaryl (optionally substituted by one or more substituents selected from Z1);
B2 represents aryl optionally substituted by one or more substituents selected from Y2;
A5 represents a direct bond or, preferably, C1-2 alkylene (e.g. methylene);
B5 represents aryl (e.g. phenyl), which group is preferably unsubstituted;
R10 represents hydrogen, C1-12 (e.g. C1-6, such as C1-3) alkyl (e.g. methyl; optionally substituted by one or more, e.g. one, substituent(s) selected from X8) or -A9-B9;
R11 represents hydrogen or -A9-B9;
X8 represents aryl (e.g. phenyl), which group is optionally substituted by one or more T1 substituents, but is preferably unsubstituted;
A9 represents a direct bond or C1-3 (e.g. C1-2) alkylene (e.g. methylene);
B9 represents aryl (e.g. phenyl; optionally substituted by one or more substituents selected from Y1), heteroaryl (optionally substituted by one or more substituents selected from Z1) or heterocyclyl (optionally substituted by one or more substituents selected from Z2);
X11 represents —OR14 (in which R14 preferably represents hydrogen);
R12 represents a direct bond or —CH2—;
Y1 and Y2 independently represent G1 or G2;
Z2 represents G2 or -Ax-By;
Z1 represents G1;
G1 represents —CN, —NO2, halo (e.g. fluoro or chloro), C1-4 alkyl (e.g. tert-butyl, isopropyl or methyl), which alkyl group is optionally substituted by one or more substituents selected from T5;
G2 represents -Ax-Bx, —R15—OR14 or —R15—N(R14)2;
R15 represents a direct bond;
R14 represents hydrogen, -Ax1-Bx1 or C1-4 (e.g. C1-2) alkyl, which alkyl group is optionally substituted by one or more substituents selected from E1;
Ax and Ax1 independently represent a direct bond;
Bx represents aryl (e.g. phenyl) or heteroaryl (e.g. a 5- or 6-membered heterocyclyl ring), both of which are optionally substituted as defined herein, but are more preferably unsubstituted;
By represents heterocyclyl (e.g. a 4- to 6-membered heterocyclyl group containing one or two heteroatoms preferably selected from oxygen or, more particularly, nitrogen, so forming for example a pyrrolidinyl or imidazolyl group);
Bx1 represents aryl (e.g. phenyl) optionally substituted by one or more halo (e.g. chloro) atoms;
T5 represents halo (e.g. fluoro) or —OC1-6 alkyl (e.g. —OC1-3 alkyl, such as —OCH2CH3; which alkoxy group is preferably unsubstituted);
E1 represents halo (e.g. fluoro) or —N(R18)2;
R18 represents methyl.
Particularly preferred compounds of the invention (e.g. when r represents 1) include those in which:
q represents 2, or, preferably, 1 (e.g. there is at least one R1 substituent present); when q represents 1, then (R1)q is in the 2-, 3- or 4-position;
R1 represents C1-12 (e.g. C1-6) alkyl (optionally substituted by one or more substituents selected from X1), —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10;
R2 represents hydrogen or -A2-B2;
R3 represents aryl (e.g. phenyl), —OR4, —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10;
when R1 represents —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10, then R3 preferably represents —OR4;
when R3 represents —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10, then R1 preferably represents C1-12 (e.g. C1-6) alkyl optionally substituted by one or more substituents selected from X1;
for example when r represents 1, then at least one of R1 and R3 represents —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10;
R4 represents C4-6 cycloalkyl (e.g. cyclopentyl), a 4- to 6-membered heterocyclyl group, or C1-4 alkyl (e.g. methyl, isopropyl, n-propyl, ethyl or, preferably, methyl);
R5 represents hydrogen, isobutyl, benzyl, —C(O)CH3, —C(O)Ph or —CH2—C(O)NH2.
Preferred substituents that R1 or R3 may represent (e.g. when r represents 1) include —O—CH2—C(O)NH2, —OCH2C(O)OH,
Further preferred compounds of the invention (include those in which:
for example, when R3 represents —OR4 in which R4 represents -A4-B4, A4 represents a direct bond and B4 represents a 3-tetrahydrofuranyl group, then the chiral atom of that group is preferably in the (R)-configuration.
Particularly preferred compounds of the invention (for example when r represents 1, and at least one of R1 and R3 represents —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10) include:
Further particularly preferred compounds of the invention (for example when r is is other than 1) include:
Particularly preferred compounds of the invention, for example when r represents 1) include:
Compounds of formula I may be prepared by:
(i) reaction of a compound of formula II:
or protected derivatives thereof, wherein R2, R3, R4, R5, R6, R7, m and n are as hereinbefore defined, with a compound of formula III,
wherein L1 represents a suitable leaving group, such as a sulfonate group or, more preferably an iodo, bromo or chloro group, and R1, q, r and R8 are as hereinbefore defined, in the presence of a base, such as a strong base, for instance an organometallic (e.g. organolithium) base (such as n-BuLi, s-BuLi, t-BuLi, lithium 2,2,6,6-tetramethylpiperidine or, preferably, lithium diisopropylamide) or an alkali metal-based base such as NaH and/or KO-tert-butyl. When an organolithium base is employed, it is optionally in the presence of an additive (for example, a lithium co-ordinating agent such as an ether (e.g. dimethoxyethane) or an amine (e.g. tetramethylethylenediamine (TMEDA), (−)sparteine or 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU) and the like)), for example in the presence of a suitable solvent, such as a polar aprotic solvent (e.g. tetrahydrofuran or diethyl ether), at sub-ambient temperatures (e.g. 0° C. to −78° C.) under an inert atmosphere. The skilled person will appreciate that the base may need to be added to the compound of formula II before the addition of the compound of formula III;
(ii) for compounds of formula I in which either at least one R1 represents —R12—O—R9—C(O)N(R10)R11, —R12—O—R9—C(O)OR10 or —R12—OS(O)2R10 (in which case q is other than 0), or R3 represents —R12—O—R9—C(O)N(R10)R11 or —R12—O—R9—C(O)OR10, reaction of a corresponding compound of formula I in which either R1 represents —R12—OR10 in which R10 represents hydrogen, or, R3 represents —OR4 in which R4 represents hydrogen, with a compound of formula IV,
Za-L1a IV
wherein L1a represents a suitable leaving group, such as one hereinbefore defined in respect of L1 (and preferably L1a represents iodo, chloro, bromo or a sulfonate group), Za represents —R9—C(O)N(R10)R11, R9—C(O)OR10 or —S(O)2R10 (as appropriate), in which R10 and R11 are as hereinbefore defined, and preferably other than hydrogen, and R9 is as hereinbefore defined, for example at around room temperature or above (e.g. up to 40-180° C.), optionally in the presence of a suitable base (e.g. sodium hydride, sodium bicarbonate, potassium carbonate, pyrrolidinopyridine, pyridine, triethylamine, tributylamine, trimethylamine, dimethylaminopyridine, diisopropylamine, diisopropylethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene, sodium hydroxide, N-ethyldiisopropylamine, N-(methylpolystyrene)-4-(methylamino)pyridine, potassium bis(trimethylsilyl)-amide, sodium bis(trimethylsilyl)amide, potassium tert-butoxide, lithium diisopropylamide, lithium 2,2,6,6-tetramethylpiperidine or mixtures thereof) and an appropriate solvent (e.g. tetrahydrofuran, pyridine, toluene, dichloromethane, chloroform, acetonitrile, dimethylformamide, trifluoromethylbenzene, dioxane, triethylamine, water or mixtures thereof);
(iii) for compounds of formula I in which there is an R1 substituent present that represents —R12—O—R9—C(O)N(R10)R11, reaction of a corresponding compound of formula I in which R1 represents —R12—O—R9—C(O)OR10 with a compound of formula V,
HN(R10)R11 V
wherein R10 and R11 are as hereinbefore defined, under standard amide coupling reaction conditions, for example in the presence of a suitable coupling reagent (e.g. 1,1′-carbonyldiimidazole, N,N′-dicyclohexylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (or hydrochloride thereof), N,N′-disuccinimidyl carbonate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, benzotriazol-1-yloxytris-pyrrolidinophosphonium hexafluorophosphate, bromo-tris-pyrrolidinophosphonium hexafluorophosphate, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetra-fluorocarbonate, 1-cyclohexyl-carbodiimide-3-propyloxymethyl polystyrene, O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate and/or O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate), optionally in the presence of a suitable base (e.g. sodium hydride, sodium bicarbonate, potassium carbonate, pyridine, triethylamine, dimethylaminopyridine, diisopropylamine, sodium hydroxide, potassium tert-butoxide and/or lithium diisopropylamide (or variants thereof), an appropriate solvent (e.g. tetrahydrofuran, pyridine, toluene, dichloromethane, chloroform, acetonitrile, dimethylformamide, trifluoromethylbenzene, dioxane or triethylamine) and a further additive (e.g. 1-hydroxybenzotriazole hydrate). Alternatively, the carboxylic acid group of the compound of formula VI may be converted under standard conditions to the corresponding acyl chloride (e.g. in the presence of SOCl2 or oxalyl chloride), which acyl chloride is then reacted with a compound of formula VII, for example under similar conditions to those mentioned above;
(iv) for compounds of formula I in which R2 represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X2) or -A2-B2 (in which A2 represents optionally substituted C1-12 alkylene), reaction of a corresponding compound of formula I in which R2 represents hydrogen, with a compound of formula VI,
Zb-L1b VI
wherein Zb represents C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X2) or -A2a-B2, in which A2a represents A2 provided that it does not represent a direct bond, and L1b represents a suitable leaving group, such as one hereinbefore defined in respect of L1, under standard reaction conditions, for example such as those hereinbefore described in respect of process step (i) above;
(v) for compounds of formula I in which there is an R1 substituent present that represents —R12—C(═NR10)N(R10)R11 or —R12—C(═NOH)N(R10)R11, in which R10 and
R11 represent hydrogen, may be prepared by reaction of a corresponding compound of formula I in which R1 represents —R12—CN, for example in the presence of a suitable amidating (or hydroxyamidating) reagent, such as hydroxylamine (or a suitable derivative thereof, such as an acetyl protected derivative thereof), optionally in the presence of a suitable solvent (such as an alcoholic solvent, e.g. ethanol) and optionally (e.g. in the case of preparation of the amidino substituent) in the presence of an acid (e.g. a hydrogen halide, e.g. HCl), which reaction may be performed at reflux, (e.g. in the case of preparation of the hydroxyamido substituent);
(vi) for compounds of formula I in which q is other than 0, and R1 represents -A1-B1 (and A1 represents a direct bond), —R12—N(R10)R11, —R12—OR10, —R12—OC(O)R10, —R12—C(O)OR10, —R12—N(R10)C(O)N(R10)R11, —R12—O—R9—C(O)OR10, —R12—O—R9—C(O)N(R10)R11, —R12—S(O)pxR10 (in which px represents 0), —R12—OS(O)2R10, —R12—N(R10)S(O)txN(R10)R11, —R12—B(OR10)2, —R12—P(R10)R11 or —R12—P(O)(OR10)2, and R12 represents, preferably at each occurrence, a direct bond, reaction of a compound of formula VII,
wherein L1c represents a suitable leaving group, such as chloro, bromo, iodo, a sulfonate group (e.g. —OS(O)2 CF3, —OS(O)2 CH3, —OS(O)2 PhMe or a nonaflate), —B(OH)2, —B(ORwx)2, —Sn(Rwx)3 or diazonium salts, in which each Rwx independently represents a C1-6 alkyl group, or, in the case of —B(ORwx)2, the respective Rwx groups may be linked together to form a 4- to 6-membered cyclic group (such as a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group), and preferably L1 represents bromo or a sulfonate group such as trifluoromethanesulfonate group, q1 represent q, provided that it does not represent 0, and R2, R3, R4, R5, R6, R7, R8, m, n and r are as hereinbefore defined, either:
Zd-H VIII
wherein q1, R2, R3, R4, R5, R6, R7, R8, m, n and r are as hereinbefore defined, with a compound of formula V as hereinbefore defined, for example under reaction conditions such as those hereinbefore described in respect of process step (i) above;
(viii) intramolecular cyclisation of a compound of formula X,
or a protected derivative thereof (e.g. an ester derivative such as a tert-butyl ester thereof), wherein R1, R2, R3, R4, R6, R7, R8, m, n, q and r are as hereinbefore defined (the skilled person will appreciate that —(R6)m represents five optional R6 substituents situated at the five free positions α, β and γ to the requisite —NH2 group), under standard conditions, for example in the presence of a suitable base (such as an alkali metal base, e.g. NaOH, or an organic base, such as an amine base, e.g. triethylamine) an appropriate solvent (such as a polar solvent, e.g. dimethylformamide, or a solvent such as an alcohol, e.g. MeOH, which may have been employed in acid, e.g. para-toluene sulfonic acid, to promote a trans-esterification or esterification of a compound of formula VIIC to form a methyl ester intermediate compound of formula VIIC). Such a process for the preparation of compounds of formula I, as well as alternative processes, are fully described in international patent application WO 2004/031149 and U.S. Pat. No. 6,770,658, the disclosures of which are incorporated in full by reference herein. Alternatively, similar reaction conditions to those described hereinbefore with reference to process step (iii) may be employed;
(ix) reaction of a compound of formula XI,
wherein L2 represents a suitable leaving group such as chloro, bromo, iodo, a sulfonate group (e.g. —OS(O)2 CF3, —OS(O)2 CH3, —OS(O)2 PhMe or a nonaflate), —B(OH)2, —B(ORwx)2, —Sn(Rwx)3 or diazonium salts, in which each Rwx independently represents a C1-6 alkyl group, or, in the case of —B(ORwx)2, the respective Rwx groups may be linked together to form a 4- to 6-membered cyclic group (such as a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group), and L2 preferably represents bromo, and R3, R4, R7 and n are as hereinbefore defined, with a compound of formula XII,
or a tautomer thereof or derivative thereof (including a possible derivative of the tautomer, e.g. a hydroxy protected tautomer, or a (1N)-protected derivative), wherein L3 represents a suitable leaving group, such as —B(OH)2 or a protected derivative thereof, for example a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group, 9-borabicyclo[3.3.1]-nonane (9-BBN), —Sn(alkyl)3 (e.g. —SnMe3 or —SnBu3), or a similar group known to the skilled person and L3 preferably represents —B(OH)2 (the skilled person will also appreciate that L2 and L3 should be mutually compatible, and may also be interchanged), and R1, R2, R5, R6, R8, L3 m and r are as hereinbefore defined, for example under conditions such as those hereinbefore defined in respect of process step (vi) above;
(x) for compounds of formula I in which R2 represents hydrogen, and there is a maximum of two R6 substituents present (at the 4- and/or 6-position), reduction of a compound of formula XIII,
or a tautomer or protected derivative thereof (e.g. a protected hydroxy tautomer, or, a compound protected at the (1N)-position), wherein m1 represents 0, 1 or 2 (and a single R6 group is therefore possible at the 4- and/or 6-position of the piperidin-2-one ring), and R1, R3, R4, R5, R6, R7, R8, q, r and n are as hereinbefore defined, for example under standard conditions, such as in the presence of a suitable reducing agent such as NaBH4 (e.g. in the presence of a suitable additive), LiAlH4 or under hydrogenation reaction conditions (e.g. catalytic hydrogenation conditions in the presence of a precious metal catalyst, e.g. Pd/C);
(xi) for compounds of formula I in which R3 represents —OR4 in which R4 is other than hydrogen, or for compounds of formula I in which R4 (at the position para to the point of attachment of the piperidin-2-one ring) is other than hydrogen, reaction of a corresponding compound of formula I in which R3 represents —OH or,
R4 represents hydrogen, with a compound of formula XIV,
R4a-L2x XIV
wherein R4a represents —R9—OR10, —R9—C(O)OR10, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl (which latter three groups are optionally substituted by one or more substituents selected from X3) or -A4-B4, L2x represents a suitable leaving group such as one defined hereinbefore in respect of L2, and R9, R10, X3, A4 and B4 are as hereinbefore defined, under standard reaction conditions, for example such as those hereinbefore described in respect of process step (ii) or (vi). The skilled person will appreciate that when the reaction is with a compound of formula XIV in which L2x is e.g. bromo, chloro or a sulfonate group, then the conditions described in process step (ii) are preferably employed, whereas for reaction with a compound of formula XIV in which L2x is —B(OH)2, —B(ORwx)2 or —Sn(Rwx)3, then the reaction is preferably preformed under the reaction conditions described in process step (vi);
(xii) for compounds of formula I in which R3 represents —OR4 in which R4 is other than hydrogen, or for compounds of formula I in which R4 (at the position para to the point of attachment of the piperidin-2-one ring) is other than hydrogen, reaction of a compound of formula XV,
wherein L2x1 represents L2x or R3, L2x2 represents L2x or —OR4, provided that at least one of R2x1 and R2x2 represents L2x, in which L2x is as hereinbefore defined and preferably represents a suitable leaving group such as bromo, and R1, R2, R5, R6, R7, m and n are as hereinbefore defined, with a compound of formula XVI,
R4—OH XVI
wherein R4 is as hereinbefore defined, under standard reaction conditions, for example such as those hereinbefore described in respect of process (xi) (preferably with reference to process (vi));
(xiii) compounds of formula I in which R2 represents —OR4 in which R4 represents hydrogen, may be prepared by reaction of a corresponding compound of formula I in which R2 represents hydrogen, with a base (such as one described hereinbefore in respect of preparation of compounds of formula I, process step (i)), optionally in the presence of an additive and solvent (such as one hereinbefore described in respect of process step (i), for example Cu salts may be employed as the optional additive), followed by quenching with oxygen or a suitable equivalent thereof under standard conditions.
Compounds of formula II in which R2 represents —OR4 in which R4 represents hydrogen, may alternatively be prepared by reaction of a compound of formula XVI,
or a protected derivative thereof (e.g. protected at the (1N)-position), wherein each Rz independently represents C1-6 alkyl (e.g. methyl, so forming for example a trimethylsilyl group), and R3, R4, R5, R6, R7, m and n are as hereinbefore defined, under double bond epoxidation reaction conditions known to those skilled in the art, for example in the presence of a suitable oxidising reagent such as meta chloro perbenzoic acid (mcpba). The skilled person will appreciate that an epoxide intermediate may be formed, which may not be stable and thus may hydrolyse during work-up to form the relevant compound of formula I (alternatively, the intermediate so formed may be deprotected e.g. under mild acidic conditions, or in the presence of fluoride ions, in order to promote the formation of the relevant compound of formula II).
Compounds of formula II in which R2 represents —OR4 in which R4 represents hydrogen, may alternatively be prepared by reaction of a compound of formula
or a protected derivative thereof (e.g. protected at the (1N)-position), wherein Rz, R3, R4, R5, R6, R7, m and n are as hereinbefore defined, in the presence of a suitable oxidising agent, and under reaction conditions such as those hereinbefore described (e.g. the oxidation conditions employed for the synthesis of compounds of formula II).
Compounds of formula II in which m represents 0, 1 or 2 (forming a compound of formula II in which a R6 group is optionally present at the 4- and/or 6-position of the piperidin-2-one) may be prepared by reduction of a compound of formula XVIII,
or a tautomer or protected derivative thereof (e.g. a protected hydroxy tautomer, or, a compound protected at the (1N)-position), wherein m1, R2, R3, R4, R5, R6, R7 and n are as hereinbefore defined, for example under reaction conditions such as those hereinbefore described in respect of preparation of compounds of formula I (process step (x) above).
Compounds of formula III in which L1 represents bromo may be prepared by reaction of a compound corresponding to a compound of formula III but in which L1 represents —OH, under standard reaction conditions that may effect such a conversion, for example conversion to an appropriate sulfonate group followed by reaction in the presence of bromide ions (or a suitable source thereof). The reaction may also be performed in the presence of an appropriate brominating reagent (e.g. in the presence of PBr3 or CBr4, optionally in the presence of PPh3, and optionally in the presence of a suitable solvent such as dichloromethane or diethyl ether).
Compounds of formula XVI may be prepared by reaction of a corresponding compound of formula II in which R2 represents hydrogen (or a suitable protected derivative thereof, such as a (1N)-protected derivative), with an appropriate trialkylsilyl chloride (e.g. trimethylsilyl chloride), or the like, under standard reaction conditions, for example such as those hereinbefore described in respect of preparation of compounds of formula I (process step (ii) above).
Compounds of formula XVII may be prepared from corresponding compounds of formula X in which R1 and R2 both represent hydrogen, (or a suitable protected derivative thereof, such as a (1N)-protected derivative), with an appropriate trialkylsilyl chloride (e.g. trimethylsilyl chloride), or the like, under standard reaction conditions.
Compounds of formula XIII and XVIII may be prepared by reaction of a compound of formula XI as hereinbefore defined, with a compound of formula XIX,
or a tautomer thereof or derivative thereof (including a possible derivative of the tautomer, e.g. a hydroxy protected tautomer, or, a protected derivative, such as a (1N)-nitrogen protected derivative, e.g. N-benzyl-2-piperidinone, or, 2-methoxypyridine or 2-chloropyridine), wherein R1/2 represents R2 (for the preparation of compounds of formula XVIII), or:
(for the preparation of compounds of formula XIII), and L3, m1, R1, R2, R5, R6, R8, r and q are as hereinbefore defined, for example under conditions such as those hereinbefore described in respect of preparation of compounds of formula I (process step (vi) above). Alternatively, the values of L2 and L3 in the compounds of formula VIII and XVB may be interchanged.
Alternatively, compounds of formula XIII, or compounds of formula XVIII in which R2 represents C1-12 alkyl, C2-12 alkenyl or C2-12 alkynyl, all of which are optionally substituted as hereinbefore defined, may be prepared by reaction of a compound of formula XX,
wherein L2a represents a suitable leaving group, such one hereinbefore defined in respect of L2, and m1, n, R3, R4, R5, R6 and R7 are as hereinbefore defined, with a compound of formula XXI,
R1/2a-L3a XXI
wherein L3a represents a suitable leaving group, such as one hereinbefore defined in respect of L3 (or, alternatively, the definitions of L2a and L3a may be interchanged), R1/2a represents R2, provided that it does not represent hydrogen or —OR4 (for the preparation of the relevant compounds of formula XVIII) or:
(for the preparation of the relevant compounds of formula XIII), for example under conditions such as those hereinbefore described in respect of preparation of compounds of formula I (process step (vi) above).
For compounds of formula XVIII in which R2 represents —OR4, reaction with a compound of formula XX as hereinbefore defined, with a compound of formula XVI as hereinbefore defined, under standard reaction conditions, for example under conditions such as those hereinbefore described in respect of preparation of compounds of formula I (process step (vi) above).
Alternatively, compounds of formula XIII or compounds of formula XVIIII in which m1 represents 1 or 2, may be prepared by reaction of a compound of formula XXII,
wherein L2b represents a suitable leaving group, such as one hereinbefore defined in respect of L2, m2 represents 1 or 2 (and hence there are one or two L2b groups present at the 4- and/or 6-position of the 2-pyridinone ring), and R3, R4, R5, R7, R1/2 and n are as hereinbefore defined, with a compound of formula XXIII,
R6-L3b XXIII
wherein L3b represents a suitable leaving group, such as one hereinbefore defined in respect of L3 (or alternatively, the values of L2b and L3b may be interchanged), and R6 is as hereinbefore defined, under standard reaction conditions, for example under conditions such as those hereinbefore described in respect of preparation of compounds of formula I (process step (vi) above).
Compounds of formulae IV, V, VI, VII, VIII, IX, X, XI, XII, XIV, XV, XIX, XX, XXI, XXII and XXIII (and also e.g. certain compounds of formulae II and III) may be commercially available, are known in the literature, or may be obtained either by analogy with the processes described herein, or by conventional synthetic procedures, in accordance with standard techniques, from available starting materials using appropriate reagents and reaction conditions. In this respect, the skilled person may refer to inter alia “Comprehensive Organic Synthesis” by B. M. Trost and I. Fleming, Pergamon Press, 1991. Further, compounds of formula II may also be prepared in accordance with synthetic routes and techniques described in international patent applications WO 2004/031149 and WO 00/14083 and/or U.S. Pat. No. 6,770,658.
The substituents L1, R1, R2, R3, R4, R5, R6, R7 and R8 either in final compounds of the invention or in relevant intermediates (as appropriate) may be modified one or more times, after or during the processes described above by way of methods that are well known to those skilled in the art. Examples of such methods include substitutions, reductions, oxidations, alkylations, acylations, hydrolyses, esterifications, etherifications, halogenations or nitrations. Such reactions may result in the formation of a symmetric or asymmetric final compound of the invention or intermediate. In this respect, the skilled person may also refer to “Comprehensive Organic Functional Group Transformations” by A. R. Katritzky, O. Meth-Cohn and C. W. Rees, Pergamon Press, 1995. Specific transformation steps that may be mentioned include the conversion of one L1 group (in the compound of formula III) into another L1 group (e.g. the conversion of one halo group, such as chloro, into another halo group, such as iodo, for example by reaction in the presence of potassium iodide), or even the conversion of a hydroxy group to a L1 group. Other transformation steps include the reduction of a nitro group to an amino group, the reduction of a cyano group to a methylamino group, the hydrolysis of a nitrile group to a carboxylic acid group, and standard nucleophilic aromatic substitution reactions.
It will also be appreciated by those skilled in the art that in the process described below the functional groups of intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxy, amino, mercapto and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, methyl and the like. Suitable protecting groups for amino, amidino and guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include —C(O)—R″ (where R″ is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl or aralkyl esters.
Protecting groups may be added or removed in accordance with standard techniques (for example a methyl protecting group on a hydroxy group may be removed by reaction in the presence of a suitable ‘cleaving reagent’ such as BBr3), which are known to one skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T. W. and P. G. M. Wuts, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley.
The protecting group may also be a polymer resin such as a Wang resin or a 2-chlorotrityl-chloride resin.
It will also be appreciated by those skilled in the art, although such protected derivatives of compounds of this invention may not possess pharmacological activity as such, they may be administered to a mammal and thereafter metabolised in the body to form compounds of the invention which are pharmacologically active. Such derivatives may therefore be described as “prodrugs”. All prodrugs of compounds of this invention are included within the scope of the invention.
Compounds of the invention are indicated as pharmaceuticals. According to a further aspect of the invention there is provided a compound of the invention, as hereinbefore defined, for use as a pharmaceutical.
According to a further aspect of the invention there is provided a pharmaceutical composition/formulation including a compound of the invention, as hereinbefore defined, in admixture with a pharmaceutically acceptable adjuvant, carrier, diluent or excipient.
Preferred pharmaceutical formulations include those in which the active ingredient is present in at least 1% (such as at least 10%, preferably in at least 30% and most preferably in at least 50%) by weight. That is, the ratio of active ingredient to the other components (i.e. the addition of adjuvant, diluent and carrier) of the pharmaceutical composition is at least 1:99 (e.g. at least 10:90, preferably at least 30:70 and most preferably at least 50:50) by weight.
Such compositions/formulations may be prepared in accordance with standard and/or accepted pharmaceutical practice.
Compounds of the invention may be useful in treating or preventing inflammatory diseases or conditions in a patient. Hence, in another aspect, this invention is directed to methods for treating or preventing an inflammatory disease or condition in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a compound of the invention as hereinbefore described or a therapeutically effective amount of a pharmaceutical formulation/composition of the invention as hereinbefore described.
The term “inflammation” will thus also be understood to include any inflammatory disease, disorder or condition per se, any condition that has an inflammatory component associated with it, and/or any condition characterised by inflammation as a symptom. Accordingly, compounds of the invention may be useful in the treatment of the inflammatory diseases or conditions described herein, and/or (if appropriate) inflammation that may be associated with such diseases or conditions.
The inflammatory condition or disease may be an autoimmune condition or disease; the inflammatory condition or disease may involve acute or chronic inflammation of bone and/or cartilage compartments of joints; the inflammatory condition or disease may be an arthritis selected from rheumatoid arthritis, gouty arthritis or juvenile rheumatoid arthritis; the inflammatory condition or disease may be a respiratory disorder selected from asthma or a chronic obstructive pulmonary disease (COPD, e.g., emphysema or chronic bronchitis); the condition or disease may be associated with the disregulation of T-cells; the condition or disease may be associated with elevated levels of inflammatory cytokines (e.g., wherein the inflammatory cytokine is IL-2, or wherein the inflammatory cytokine is IFN-γ, or wherein the inflammatory cytokine is TNF-α); the inflammatory condition or disease may be multiple sclerosis; the inflammatory condition or disease may be pulmonary sarcadosis; the inflammatory condition or disease may be ocular inflammation or allergy; the inflammatory condition or disease may be an inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis); and the inflammatory condition or disease may be an inflammatory cutaneous disease (e.g., psoriasis or dermatitis).
Compounds of the invention may be useful in modulating intracellular cyclic adenosine 5′-monophosphate levels within a mammal, preferably a human, Hence, in another aspect, this invention is directed to methods for modulating intracellular cyclic adenosine 5′-monophosphate levels within a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof an amount of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described effective to modulate the intracellular cyclic adenosine 5′-monophosphate levels of the mammal. The mammal, preferably a human, may have an inflammatory condition or disease (for example one defined herein).
Compounds of the invention may be useful in treating or preventing a disease or condition in a mammal, preferably a human, where the disease or condition is associated with pathological conditions that are modulated by inhibiting enzymes associated with secondary cellular messengers. Hence, in another aspect, this invention is directed to methods for treating or preventing a disease or condition in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described, and the disease or condition is associated with pathological conditions that are modulated by inhibiting enzymes associated with secondary cellular messengers. Such enzymes (that may be inhibited) may be a cyclic AMP phosphodiesterase; a phosphodiesterase 4; a phosphodiesterase 3; or a cyclic GMP phosphodiesterase. Further, more than one type of enzyme may be inhibited, for instance, the enzymes may be both phosphodiesterase 4 and phosphodiesterase 3.
Compounds of the invention may be useful in treating or preventing uncontrolled cellular proliferation in a mammal, preferably a human. Hence, in another aspect, this invention is directed to methods for treating or preventing uncontrolled cellular proliferation in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount (e.g. an amount effective to treat or prevent uncontrolled cellular) of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described. The uncontrolled cellular proliferation may be caused by a cancer selected from leukaemia and solid tumors.
Compounds of the invention may be useful in treating or preventing transplant rejection in a mammal, preferably a human. Hence, in another aspect, this invention is directed to methods for treating or preventing transplant rejection in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount (e.g. an amount effective to treat or prevent transplant rejection in the mammal) of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described. The rejection may be due to graft versus host disease.
Compounds of the invention may be useful in treating or preventing conditions associated with the central nervous system (CNS) in a mammal, preferably a human. Hence, in another aspect, this invention is directed to methods for treating or preventing conditions associated with the central nervous system in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount (e.g. an amount effective to treat or prevent conditions associated with the central nervous system (CNS) in the mammal) of a compound of the invention as described above or a pharmaceutical formulation/composition of the invention as hereinbefore described. The condition associated with the central nervous system (CNS) may be depression.
These and other aspects and embodiments of the present invention will be apparent upon reference to the following detailed description. To this end, various references are set forth herein which describe in more detail certain procedures, compounds and/or formulations/compositions, and are hereby incorporated by reference in their entirety.
In a method of the present invention, a compound of the invention (as hereinbefore defined), or a formulation/composition comprising one or more compounds of the invention (as hereinbefore defined) in admixture with a pharmaceutically acceptable adjuvant, carrier, diluent or excipient, may, although need not, achieve one or more of the following desired results in a subject to whom has been administered a compound of the invention as hereinbefore defined, or a formulation/composition containing one of these compounds and a pharmaceutically acceptable adjuvant, carrier, diluent or excipient:
1. Inhibition of reactive oxygen species generation from primary neutrophils;
2. Inhibition of neutrophil chemotaxis;
3. Inhibition of TNF-α production;
4. Inhibition of edema;
5. Oxygen radical scavenging;
6. Inhibition of cyclic-AMP phosphodiesterases 1, 3 and/or 4, and related PDEs such as PDE7;
7. Potentiate induction of CRE-mediated transcription activity in human monocytic cells;
8. Inhibition of PDE, preferably PDE4, PDE3, or PDE3 and PDE4;
9. Inhibition of cytokine production by activated T-cell subsets;
10. Inhibition of neutrophil myeloperoxidase release;
11. Low ratio of IC50 PDE4(cat):IC50PDE4(HARBS);
12. Inhibition of graft rejection;
13. Inhibition of clinical and histopathological parameters of disease in inflammatory bowel disease;
14. Inhibition of clinical and histopathological parameters of arthritis in a murine collage-induced arthritis model;
15. Inhibition of clinical and histopathological parameters of disease in asthma; and
16. Inhibition of clinical and histopathological parameters of disease in COPD.
Thus, the compounds and compositions of the invention may be used to treat inflammation, including both acute and chronic inflammation as well as certain proliferative disorders (cancers). As used herein, inflammation includes, without limitation, ankylosing spondylitis, arthritis (where this term encompasses over 100 kinds of rheumatic diseases), asthma, chronic bronchitis, Crohn's disease, fibromyalgia syndrome, gout, inflammations of the brain (including multiple sclerosis, AIDS dementia, Lyme encephalopathy, herpes encephalitis, Creutzfeld-Jakob disease, and cerebral toxoplasmosis), emphysema, inflammatory bowel disease, irritable bowel syndrome, ischemia-reperfusion injury juvenile erythematosus pulmonary sarcoidosis, Kawasaki disease, osteoarthritis, pelvic inflammatory disease, psoriatic arthritis (psoriasis), rheumatoid arthritis, psoriasis, tissue/organ transplant, scleroderma, spondyloarthropathies, systemic lupus erythematosus, pulmonary sarcoidosis, and ulcerative colitis. As used herein, proliferative disorders includes, without limitation, all leukemias and solid tumors that are susceptible to undergoing differentiation or apoptosis upon interruption of their cell cycle.
The compounds of the invention may be tested for the above indications in the assays described below in the Biological Examples. In addition, the compounds of the invention may be tested in animal models to further demonstrate their enzymatic, cellular, anti-inflammatory and central nervous system activity. Specifically, the compounds of the invention may be tested in animal models for diseases and pathological conditions of the central nervous system, including, but not limited to, cognitive function, Alzheimer's disease, learning and memory (Rose et al., “Phosphodiesterase inhibitors for cognitive enhancement,” Curr. Pharm. Des. 2005; 11(26):3329-34), Rubinstein Taybi Syndrome (Bourtchouladze et al., “A mouse model of Rubinstein-Taybi syndrome: Defective long term memory is ameliorated by inhibitors of phosphodiesterase 4, Proc. Natl. Acad. Sci. USA. (2003), September 2; 100(18):10518-22.), cerebrovascular disease, depression (Zhu et al., “The antidepressant and antiinflammatory effects of rolipram in the central nervous system”, CNS Drug Rev. (2001);7(4):387-98), schizophrenia, Parkinson's disease (Weishaar et al., “A new generation of phosphodiesterase inhibitors: multiple molecular forms of phosphodiesterase and the potential for drug selectivity”, J Med Chem. (1985), May; 28(5):537-45.), multiple sclerosis (Huang et al., “The next generation of PDE4 inhibitors”, Curr Opin Chem Biol. (2001), August; 5(4):432-8; Dyke and Montana, “Update on the therapeutic potential of PDE4 inhibitors”, Expert Opin Investig Drugs. (2002), January; 11(1):1-13) and allergic rhinitis. In addition, the compounds of the invention may be tested in animal models for inflammatory and immune disorders or pathological conditions including, but not limited to, cancer (Weishaar et al., 1985), asthma (Huang et al., 2001; Dyke and Montana, 2002), chronic obstructive pulmonary disease (Huang et al., 2001; Dyke and Montana, 2002), respiratory distress syndrome, rhinitis, nephritis, psoriasis (Houslay et al., “Keynote review: phosphodiesterase-4 as a therapeutic target”, Drug Discov Today. (2005), November 15; 10(22):1503-19), eczema, atopic dermatitis, urticaria, conjunctivitis, inflammatory bowel diseases (Huang et al., 2001), Crohn's disease, ulcerative colitis, rheumatoid arthritis (Huang et al., 2001), osteoarthritis, eosinophilic gastrointestinal disorders, vascular disease and diabetes mellitus. With respect to allergic, inflammatory and autoimmune disease, established pre-clinical models may be used and may include: hapten models of dermatitis; collagen-induced arthritis (CIA), adjuvant induced arthritis, cartilage degradation models in the mouse or rat LPS-induced joint inflammation; rat and mouse lung LPS, cytokine, allergen and cigarette smoke-mediated inflammation, lung function and airway remodeling models such as rat tracheal explant model; dextran sodium sulphate (DSS) and trinitrobenzenesulfonic-acid (TNBS) induced colitis in the mouse and rat; behavioral models of learning and memory such as object recognition, fear conditioning, Morris water escape task, passive avoidance test and radial arm maze test; behavioral models of depression such as chronic stress test, tail suspension test, forced swim test, reserpine-mediated hypothermia and yohimbine-induced lethality test.
Compounds of the invention may inhibit disease induction in these models at doses of less than 20 mg/kg. The Biological Examples below outline some, but not all, of the preclinical models that may be used to support the claims of this patent. For instance, compounds of the examples (described hereinafter) were tested in the Biological examples, and were found to exhibit 50% inhibition of PDE4 at a concentration of 20 μM or below (and more preferably at a concentration of 10 μM or below).
Compounds of the invention may also be combined with other therapeutic agents that are useful in the treatment of the conditions described herein. For instance, the compounds of the invention may be combined with other compounds that may be useful in the treatment of:
i) an inflammatory disorder;
ii) a disorder in which the modulation of intracellular cyclic adenosine 5′-monophosphate levels within a mammal is desired and/or required, which disorder may be an inflammatory disorder;
iii) a disorder associated with pathological conditions that are modulated by inhibiting enzymes associated with secondary cellular messengers (e.g. a cyclic AMP phosphodiesterase; a phosphodiesterase 4; a phosphodiesterase 3; a cyclic GMP phosphodiesterase; or both phosphodiesterase 4 and phosphodiesterase 3), which disorder may be an inflammatory disorder; iv) transplant rejection in a mammal;
v) uncontrolled cellular proliferation; and/or
vi) a disorder associated with the central nervous system.
According to a further aspect of the invention, there is provided a combination product comprising:
Such combination products provide for the administration of a compound of the invention in conjunction with the other therapeutic agent, and may thus be presented either as separate formulations, wherein at least one of those formulations comprises a compound of the invention, and at least one comprises the other therapeutic agent, or may be presented (i.e. formulated) as a combined preparation (i.e. presented as a single formulation including a compound of the invention and the other therapeutic agent).
Thus, there is further provided:
(1) a pharmaceutical formulation/composition including a compound of the invention, as hereinbefore defined, another therapeutic agent that is useful in the treatment of i), ii), iii), iv), v) or vi) above (e.g. a therapeutic agent that is useful in the treatment of an inflammatory disorder), and a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient; and
(2) a kit of parts comprising components:
The invention further provides a process for the preparation of a combination product as hereinbefore defined, which process comprises bringing into association a compound of the invention, as hereinbefore defined, or a pharmaceutically acceptable derivative (e.g. salt) thereof with another therapeutic agent that is useful in the treatment of i), ii), iii), iv), v) or vi) above (e.g. a therapeutic agent that is useful in the treatment of an inflammatory disorder), and at least one pharmaceutically-acceptable adjuvant, diluent, carrier or excipient.
By “bringing into association”, we mean that the two components are rendered suitable for administration in conjunction with each other.
Thus, in relation to the process for the preparation of a kit of parts as hereinbefore defined, by bringing the two components “into association with” each other, we include that the two components of the kit of parts may be:
(i) provided as separate formulations (i.e. independently of one another), which are subsequently brought together for use in conjunction with each other in combination therapy; or
(ii) packaged and presented together as separate components of a “combination pack” for use in conjunction with each other in combination therapy.
As stated above, the present invention also relates to pharmaceutical composition containing the compounds of the invention disclosed herein. In one embodiment, the present invention relates to a composition comprising compounds of the invention in a pharmaceutically acceptable carrier and in an amount effective to treat a disease or condition of interest as disclosed herein, such as inflammation and/or rheumatoid arthritis, when administered to an animal, preferably a mammal, most preferably to a human.
Administration of the compounds of the invention, or their pharmaceutically acceptable salts, in pure form or in an appropriate pharmaceutical composition, can be carried out via any of the accepted modes of administration of agents for serving similar utilities.
For instance, compounds of the invention may be administered orally, intravenously, subcutaneously, buccally, rectally, dermally, nasally, tracheally, bronchially, sublingually, by any other parenteral route or via inhalation, in a pharmaceutically acceptable dosage form.
Compounds of the invention may be administered alone, but are preferably administered by way of known pharmaceutical compositions/formulations, including tablets, capsules or elixirs for oral administration, suppositories for rectal administration, sterile solutions or suspensions for parenteral or intramuscular administration, and the like.
The invention further provides a process for the preparation of a pharmaceutical composition/formulation, as hereinbefore defined, which process comprises bringing into association a compound of the invention, as hereinbefore defined, or a pharmaceutically acceptable derivative (e.g. salt) thereof, with a pharmaceutically-acceptable adjuvant, carrier, diluent or excipient.
The pharmaceutical compositions so prepared may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. Typical routes of administering such pharmaceutical compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, rectal, vaginal, and intranasal. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques. Pharmaceutical compositions of the invention are formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. Compositions that will be administered to a subject or patient take the form of one or more dosage units, where for example, a tablet may be a single dosage unit, and a container of a compound of the invention in aerosol form may hold a plurality of dosage units. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered will, in any event, contain a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, for treatment of a disease or condition of interest in accordance with the teachings of this invention.
A pharmaceutical composition of the invention may be in the form of a solid or liquid. In one aspect, the carrier(s) are particulate, so that the compositions are, for example, in tablet or powder form. The carrier(s) may be liquid, with the compositions being, for example, an oral syrup, injectable liquid or an aerosol, which is useful in, for example, inhalatory administration.
When intended for oral administration, the pharmaceutical composition is preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension and gel forms are included within the forms considered herein as either solid or liquid.
As a solid composition for oral administration, the pharmaceutical composition may be formulated into a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like form. Such a solid composition will typically contain one or more inert diluents or edible carriers. In addition, one or more of the following may be present: binders such as carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate or orange flavoring; and a coloring agent.
When the pharmaceutical composition is in the form of a capsule, for example, a gelatin capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or oil.
The pharmaceutical composition may be in the form of a liquid, for example, an elixir, syrup, solution, emulsion or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, preferred composition contain, in addition to the present compounds, one or more of a sweetening agent, preservatives, dye/colorant and flavour enhancer.
In a composition intended to be administered by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent may be included.
The liquid pharmaceutical compositions of the invention, whether they be solutions, suspensions or other like form, may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
Physiological saline is a preferred adjuvant. An injectable pharmaceutical composition is preferably sterile.
A liquid pharmaceutical composition of the invention intended for either parenteral or oral administration should contain an amount of a compound of the invention such that a suitable dosage will be obtained. Typically, this amount is at least 0.01% of a compound of the invention in the composition. When intended for oral administration, this amount may be varied to be between 0.1 and about 70% of the weight of the composition. Preferred oral pharmaceutical compositions contain between about 4% and about 50% of the compound of the invention.
Preferred pharmaceutical compositions and preparations according to the present invention are prepared so that a parenteral dosage unit contains between 0.01 to 10% by weight of the compound prior to dilution of the invention.
The pharmaceutical composition of the invention may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment or gel base. The base, for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickening agents may be present in a pharmaceutical composition for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or iontophoresis device. Topical formulations may contain a concentration of the compound of the invention from about 0.1 to about 10% w/v (weight per unit volume).
The pharmaceutical composition of the invention may be intended for rectal administration, in the form, for example, of a suppository, which will melt in the rectum and release the drug. The composition for rectal administration may contain an oleaginous base as a suitable nonirritating excipient. Such bases include, without limitation, lanolin, cocoa butter and polyethylene glycol.
The pharmaceutical composition of the invention may include various materials, which modify the physical form of a solid or liquid dosage unit. For example, the composition may include materials that form a coating shell around the active ingredients. The materials that form the coating shell are typically inert, and may be selected from, for example, sugar, shellac, and other enteric coating agents.
Alternatively, the active ingredients may be encased in a gelatin capsule. The pharmaceutical composition of the invention in solid or liquid form may include an agent that binds to the compound of the invention and thereby assists in the delivery of the compound. Suitable agents that may act in this capacity include a monoclonal or polyclonal antibody, a protein or a liposome.
The pharmaceutical composition of the invention may consist of dosage units that can be administered as an aerosol. The term aerosol is used to denote a variety of systems ranging from those of colloidal nature to systems consisting of pressurised packages. Delivery may be by a liquefied or compressed gas or by a suitable pump system that dispenses the active ingredients. Aerosols of compounds of the invention may be delivered in single phase, bi-phasic, or tri-phasic systems in order to deliver the active ingredient(s). Delivery of the aerosol includes the necessary container, activators, valves, subcontainers, and the like, which together may form a kit. One skilled in the art, without undue experimentation may determine preferred aerosols.
The pharmaceutical compositions of the invention may be prepared by methodology well known in the pharmaceutical art. For example, a pharmaceutical composition intended to be administered by injection can be prepared by combining a compound of the invention with sterile, distilled water so as to form a solution. A surfactant may be added to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that non-covalently interact with the compound of the invention so as to facilitate dissolution or homogeneous suspension of the compound in the aqueous delivery system.
The compounds of the invention, or their pharmaceutically acceptable salts, are administered in a therapeutically effective amount, which will vary depending upon a variety of factors including the activity of the specific compound employed; the metabolic stability and length of action of the compound; the age, body weight, general health, sex, and diet of the patient; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disease or condition; and the subject undergoing therapy. Generally, a therapeutically effective daily dose is (for a 70 kg mammal) from about 0.001 mg/kg (i.e., 0.7 mg) to about 100 mg/kg (i.e., 7.0 gm); preferably a therapeutically effective dose is (for a 70 kg mammal) from about 0.01 mg/kg (i.e., 7 mg) to about 50 mg/kg (i.e., 3.5 gm); more preferably a therapeutically effective dose is (for a 70 kg mammal) from about 1 mg/kg (i.e., 70 mg) to about 25 mg/kg (i.e., 1.75 gm).
The ranges of effective doses provided herein are not intended to be limiting and represent preferred dose ranges. However, the most preferred dosage will be tailored to the individual subject, as is understood and determinable by one skilled in the relevant arts. (see, e.g., Berkow et al., eds., The Merck Manual, 16th edition, Merck and Co., Rahway, N.J., 1992; Goodmanetna., eds., Goodman and Cilman's The Pharmacological Basis of Therapeutics, 10th edition, Pergamon Press, Inc., Elmsford, N.Y., (2001); Avery's Drug Treatment: Principles and Practice of Clinical Pharmacology and Therapeutics, 3rd edition, ADIS Press, LTD., Williams and Wilkins, Baltimore, Md. (1987), Ebadi, Pharmacology, Little, Brown and Co., Boston, (1985); Osolci al., eds., Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, Pa. (1990); Katzung, Basic and Clinical Pharmacology, Appleton and Lange, Norwalk, Conn. (1992)).
The total dose required for each treatment can be administered by multiple doses or in a single dose over the course of the day, if desired. Generally, treatment is initiated with smaller dosages, which are less than the optimum dose of the compound.
Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. The diagnostic pharmaceutical compound or composition can be administered alone or in conjunction with other diagnostics and/or pharmaceuticals directed to the pathology, or directed to other symptoms of the pathology. The recipients of administration of compounds and/or compositions of the invention can be any vertebrate animal, such as mammals. Among mammals, the preferred recipients are mammals of the Orders Primate (including humans, apes and monkeys), Arteriodactyla (including horses, goats, cows, sheep, pigs), Rodenta (including mice, rats, rabbits, and hamsters), and Carnivora (including cats, and dogs). Among birds, the preferred recipients are turkeys, chickens and other members of the same order. The most preferred recipients are humans.
For topical applications, it is preferred to administer an effective amount of a pharmaceutical composition according to the invention to target area, e.g., skin surfaces, and the like. This amount will generally range from about 0.0001 mg to about 1 g of a compound of the invention per application, depending upon the area to be treated, whether the use is diagnostic, prophylactic or therapeutic, the severity of the symptoms, and the nature of the topical vehicle employed. A preferred topical preparation is an ointment, wherein about 0.001 to about 50 mg of active ingredient is used per cc of ointment base. The pharmaceutical composition can be formulated as transdermal compositions or transdermal delivery devices (“patches”). Such compositions include, for example, a backing, active compound reservoir, a control membrane, liner and contact adhesive. Such transdermal patches may be used to provide continuous pulsatile, or on demand delivery of the compounds of the present invention as desired.
The compositions of the invention can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art. Controlled release drug delivery systems include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Examples of controlled release systems are given in U.S. Pat. Nos. 3,845,770 and 4,326,525 and in P. J. Kuzma et al, Regional Anesthesia 22 (6): 543-551 (1997), all of which are incorporated herein by reference.
The compositions of the invention can also be delivered through intra-nasal drug delivery systems for local, systemic, and nose-to-brain medical therapies. Controlled Particle Dispersion (CPD)™technology, traditional nasal spray bottles, inhalers or nebulizers are known by those skilled in the art to provide effective local and systemic delivery of drugs by targeting the olfactory region and paranasal sinuses.
The invention also relates to an intravaginal shell or core drug delivery device suitable for administration to the human or animal female. The device may be comprised of the active pharmaceutical ingredient in a polymer matrix, surrounded by a sheath, and capable of releasing the compound in a substantially zero order pattern on a daily basis similar to devises used to apply testosterone as described in PCT Patent No. WO 98/50016. Current methods for ocular delivery include topical administration (eye drops), subconjunctival injections, periocular injections, intravitreal injections, surgical implants and iontophoresis (uses a small electrical current to transportionized drugs into and through body tissues). Those skilled in the art would combine the best suited excipients with the compound for safe and effective intra-ocular administration. The most suitable route will depend on the nature and severity of the disease or condition being treated. Those skilled in the art are also familiar with determining administration methods (oral, intravenous, inhalation, subcutaneous, rectal etc.), dosage forms, suitable pharmaceutical excipients and other matters relevant to the delivery of the compounds to a subject in need thereof.
Compounds of the invention may have the advantage that they are effective inhibitors (and hence particularly effective in the treatment of the conditions described herein), and in particular effective PDE inhibitors (and especially effective PDE4 inhibitors).
Compounds of the invention may also have the advantage that they may be more efficacious than, be less toxic than, be longer acting than, be more potent than, produce fewer side effects than, be more easily absorbed than, and/or have a better pharmacokinetic profile (e.g. higher oral bioavailability and/or lower clearance) than, and/or have other useful pharmacological, physical, or chemical properties over, compounds known in the prior art, whether for use in the above-stated indications or otherwise.
All compounds of the invention as prepared above which exist in free base or acid form may be converted to their pharmaceutically acceptable salts by treatment with the appropriate inorganic or organic base or acid. Salts of the compounds prepared above may be converted to their free base or acid form by standard techniques. It is understood that all polymorphs, amorphous forms, anhydrates, hydrates, solvates and salts of the compounds of the invention are intended to be within the scope of the invention. Furthermore, all compounds of the invention which contain an ester group can be converted to the corresponding acid by methods known to one skilled in the art or by methods described herein.
The following specific Synthetic Preparations (for the preparation of starting materials and intermediates) and Synthetic Examples (for the preparation of the compounds of the invention) and the Biological Examples (for the assays used to demonstrate the utility of the compounds of the invention) are provided as a guide to assist in the practice of the invention, and are not intended as a limitation on the scope of the invention. Where one or more NMR data are given for a particular compound, each NMR may represent a single stereoisomer, a non-racemic mixture of stereoisomers or a racemic mixture of the stereoisomers of the compound.
A 2.5M solution of n-BuLi in hexanes (4.0 mL, 10 mmol) was added dropwise to a −78° C. solution of diisopropylamine (1.54 mL, 10.9 mmol) in THF (8.5 mL). After 30 minutes this 0.7M solution of LDA (9.9 mL, 7.1 mmol) was added dropwise to a −78° C. solution of (S)-tert-butyl 5-(3-(cyclopentyloxy)-4-methoxyphenyl)-2-oxopiperidine-1-carboxylate in THF (30 mL). After 30 minutes a solution of 3-benzyloxybenzylbromide (2.3 g, 8.4 mmol) in THF (10 mL) was added dropwise. The resulting solution was allowed to warm to 0° C. over 2 hours. The reaction was quenched by addition of saturated NaHCO3 solution (100 mL) then was allowed to warm to ambient temperature. The reaction mixture was extracted with 150 mL EtOAc and the extracts were washed with brine, dried over MgSO4, filtered and concentrated. Purification by column chromatography eluting with 15% EtOAc/Hexanes afforded an isomeric mixture of products (3.0 g, 84%). The product mixture was taken up in CH2Cl2 (40 mL) and was cooled to 0° C., then trifluoroacetic acid (10 mL) was added. After 1 hour the reaction mixture was diluted with 200 mL CH2Cl2 and was washed successively with water (100 mL), saturated NaHCO3 solution (100 mL) and brine (100 mL) then was dried over MgSO4, filtered and concentrated. Purification by column chromatography eluting with 30% acetone/hexanes afforded (3R,5S)-3-(3-(benzyloxy)benzyl)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)piperidin-2-one (3) (1.32 g, 53%) and (3S,5S)-3-(3-(benzyloxy)benzyl)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)piperidin-2-one (4) (0.91 g, 36%).
A solution of (3S,5S)-3-(3-(benzyloxy)benzyl)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)piperidin-2-one (4) (1.04 g, 2.15 mmol; see preparation 1 above), 10% Pd on carbon (110 mg), MeOH (6 mL) and EtOAc (9 mL) was stirred under hydrogen atmosphere at ambient temperature for 2 days. The catalyst was removed by filtration and the filtrate was concentrated to afford 823 mg (96%) of (3S,5S)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)-3-(3-hydroxybenzyl)piperidin-2-one (5).
A solution of (3-aminophenyl)methanol (10) (1.0 g, 8.1 mmol), (Boc)2O (5.3 g, 24.3 mmol), THF (25 mL) and 1M NaOH (25 mL) was stirred at ambient temperature overnight. The reaction mixture was extracted with petroleum ether and the combined extracts were dried over MgSO4, filtered and concentrated. The viscous oil was purified by column chromatography eluting with 10, 20, 40 and 100% EtOAc/hexanes to afford 1.46 g (81%) of tert-butyl 3-(hydroxymethyl)phenylcarbamate (11) as colourless, viscous oil.
A solution of tert-butyl 3-(hydroxymethyl)phenylcarbamate (11) (1.35 g, 6.05 mmol; see preparation 3 above), TBSCl (1.02 g, 6.6 mmol), DMF (12 mL) and imidazole (0.91 g, 13 mmol) was stirred at ambient temperature for 1 hour. The reaction mixture was diluted with 200 mL of ether and was washed with water and brine then was dried over MgSO4, filtered and concentrated to afford 1.91 g (94%) of tert-butyl 3-((tert-butyldimethylsilyloxy)methyl)phenylcarbamate (12) as a white solid.
tert-Butyl 3-((tert-butyldimethylsilyloxy)methyl)phenylcarbamate (12) (500 mg, 1.48 mmol; see preparation 4 above) was added to a ambient temperature solution of NaH (71 mg of a 60% dispersion in mineral oil, 1.8 mmol) and DMF (3 mL). After 40 minutes 1-iodopropane (13) (0.22 mL, 2.2 mmol) was added and the reaction was allowed to continue for 1 hour. The reaction mixture was diluted with 200 mL of ether and was washed with water and brine then was dried over MgSO4, filtered and concentrated to afford 574 mg (quantitative) of tert-butyl 3-((tert-butyldimethylsilyloxy)methyl)phenyl(propyl)carbamate (14) as a slightly yellow oil.
A solution of tert-butyl 3-((tert-butyldimethylsilyloxy)methyl)-phenyl(propyl)-carbamate (14) (574 mg, 1.48 mmol; see preparation 5 above), TBAF (1.6 mL of a 1M solution in THF, 1.6 mmol) and THF (5 mL) was heated at 65° C. for 1 hour. The reaction mixture was cooled to ambient temperature then was concentrated. The residue was purified by column chromatography eluting with 10, 20, 40 and 100% EtOAc/hexanes to afford 303 mg (75%) of tert-butyl 3-(hydroxymethyl)phenyl(propyl)carbamate (15) as a colourless oil.
A solution of tert-butyl 3-(hydroxymethyl)phenyl(propyl)carbamate (15) (300 mg, 1.13 mmol; see preparation 6 above), CBr4 (600 mg, 1.8 mmol), triphenylphosphine (356 mg, 1.4 mmol) and CH2Cl2 (6 mL) was allowed to react at ambient temperature for 1 hour. The reaction mixture was concentrated and the residue was purified by column chromatography eluting with 0, 10, 20% EtOAc/hexanes to afford 181 mg (49%) of tert-butyl 3-(bromomethyl)phenyl(propyl)carbamate (16) as a colourless oil.
In a similar manner as described above in the Synthetic Preparations, the following electrophilic intermediates were prepared for use in the preparation of the compounds of the invention:
A solution of (3S,5S)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)-3-(3-hydroxybenzyl)piperidin-2-one (857 mg, 2.17 mmol), iodoacetamide (802 mg, 4.33 mmol), K2 CO3 (898 mg, 6.50 mmol) and DMF (6 mL) was stirred at 40° C. for 18 hours. The reaction mixture was allowed to cool to ambient temperature, then was diluted with water (100 mL) and EtOAc (150 mL). The layers were separated and a white precipitate was filtered from the EtOAc layer. The aqueous layer was extracted with EtOAc and the combined EtOAc solutions were washed with saturated NaHCO3 solution and brine then dried over MgSO4, filtered and concentrated. The residue was purified by column chromatography eluting with 5%, followed by 10% MeOH/EtOAc to afford 455 mg of 2-(3-(((3S,5S)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)-2-oxopiperidin-3-yl)methyl)phenoxy)acetamide (7). The white precipitate that was isolated by filtration was dissolved in 10% MeOH/CHCl3 and was washed with saturated NaHCO3 solution and brine then dried over MgSO4, filtered and concentrated to give a total of 809 mg (82%) of 2-(3-(((3S,5S)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)-2-oxopiperidin-3-yl)-methyl)phenoxy)acetamide, MW 452.54.
A solution of 2-(((3R,5S)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)-2-oxopiperidin-3-yl)methyl)benzonitrile (8) (40 mg, 0.099 mmol), 10% Pd on carbon (11 mg), Raney-Ni (12 mg of a 50% slurry in water), LiOH.H2O (8 mg), water (0.5 mL) and 1,4-dioxane (2 mL) was stirred under a 45 psi hydrogen atmosphere at ambient temperature for overnight. The reaction mixture was eluted through a 0.45 μm filter then was diluted with CH2Cl2 (100 mL) and was washed with water and brine then dried over MgSO4, filtered and concentrated. The residue was taken up in toluene then was added a solution of 1M HCl in Et2O giving a yellow solid. The reaction mixture was briefly sonicated then was concentrated. The residue was taken up in MeOH (5 mL) then was added Si-TAAcOH (400 mg) and the resulting suspension was stirred for 1 hour then was filtered and concentrated. The residue was taken up in a minimum of CH2Cl2 then Et2O was added resulting in a white precipitate. The solvents were evaporated to afford 31 mg (70%) of (3R,5S)-3-(2-(aminomethyl)benzyl)-5-(3-(cyclopentyloxy)-4-methoxyphenyl)piperidin-2-one, MW 445.0, as a white solid.
In a similar manner as described above in the foregoing Reaction Schemes, Synthetic Preparations and Synthetic Examples; the following compounds of the invention were prepared:
Compounds of the examples inhibit disease induction in the models described herein at doses of less than 20 mg/kg. Compounds of the examples were tested in a biological test described herein, and were found to exhibit 50% inhibition of PDE4 at a concentration of 20 μM or below. For example, the following representative compounds of the examples exhibited the following IC50 values when tested in Biological Example 1, Method A:
PDE4 U937 cytoplasmic extracts were prepared by a modified procedure of the assay described in MacKenzie, S. J. and Houslay, M. D., “Action of rolipram on specific PDE4 cAMP phosphodiesterase isoforms and on the phosphorylation of cAMP-response-element-binding protein (CREB) and p38 mitogen-activated protein (MAP) kinase in U937 monocytic cells”, Biochem J. (2000), 347(Pt 2):571-8, by lysis of U937 cells (ATCC: Catalogue No. CRL-159) in M-PER Lysis buffer (Pierce) containing 10% protease inhibitor cocktail (Sigma). The cell lysates were then centrifuged at 30,000 rpm for 15 minutes at 4° C. The supernatants were aliquoted and stored at −80° C. PDE4 has been shown to be the predominant cyclic nucleotide phosphodiesterase activity in U937 cells.
An alternative source of PDE4 enzymes was from recombinant human PDE4 obtained from baculovirus-SF9 cells expression system. cDNA containing PDE4D1 was cloned into a baculovirus vector, insect cells (SF9) were then infected and cells cultured to express the PDE4 protein. The cells were lysed and used directly in assay or partially purified using standard procedures. The process can be used for other PDE4 and PDE enzymes.
Compounds of the invention were evaluated for inhibitory activity against PDE4 enzymes by the following assay Method A or B.
PDE4 assay based on modified procedure of Phosphodiesterase [3H]cAMP SPA Enzyme Assay (Amersham Biosciences, code TRKQ 7090). In this assay, PDE4 enzymes converts [3H]cAMP to [3H]5′-AMP. The assay is quenched by the addition of SPA yttrium silicate beads which preferentially bind linear nucleotides over cyclic nucleotides in the presence of zinc sulphate. The amount of [3H]5′-AMP formed is proportional to the PDE4 activity, hence PDE4 inhibitors would decrease the amount of [3H]5′-AMP formed.
Reactions were performed in duplicate by the addition of 10 μl PDE4 enzyme (U937 lysate or recombinant hPDE4) to 20 μL of assay mix and 20 μL of test compounds in Isoplates (Wallac) for 30 minutes, at 37° C. The final assay mixture contained: 50 mM Tris (pH 7.5), 8.3 mM MgCl2, 1.7 mM EGTA and [3H]cAMP (0.025 μCi) (Amersham). Assay was terminated by addition 25 μL SPA beads. The plate was sealed, shaken for 1 minute and then allowed to settle 30 minutes and the cpm determined using a Wallac Micobeta.
PDE4 assay based on modified procedure of Thompson and Appleman (Biochemistry (1971); 10; 311-316). In this assay, PDE4 enzymes converts [3H]cAMP to [3H]5′-AMP. The [3H]5′-AMP is then converted to [3H]adenosine and phosphate by nucleotidase. The amount of [3H]adenosine formed is proportional to the PDE4 activity, hence PDE4 inhibitors would decrease the amount of [3H]adenosine formed.
PDE reactions were performed for 30 minutes at 37° C. in 100 μL volumes in 1 μM cAMP, 0.05 μCi [3H]cAMP (Amersham), 0.5 U/mL 5′-nucleotidase (Sigma), 50 mM Tris, 10 mM MgCl2 pH 7.5. Reactions were performed in duplicate. Reactions were terminated by boiling for 2 minutes at 100° C. followed by the addition of 200 μL Dowex 1-8 400 Cl− anion exchange resin in a ratio of 1 resin:2 methanol:1H2O. Samples were mixed by inversion and then allowed to settle for 2-3 hours. An aliquot of 75 μL was transferred to Isoplates (Wallac), 150 μL of scintillation fluid added and the plate sealed and shaken for 30 minutes. The cpm was determined using a Wallac Micobeta.
Compounds of invention were dissolved in 100% DMSO and diluted such that the final DMSO concentration in the assay did not exceed 1% to avoid affecting the PDE4 activity. PDE4 enzyme was added in quantities such that less than 15% of substrate was consumed (linear assay conditions). Test compounds were assayed at 6-8 concentrations of ranging from 0.1 nM to 30 μM and IC50 values determined from the concentration curves by nonlinear regression analysis (GraphPad Prism® 4).
Compounds of the invention, when tested in these assays, demonstrated the ability to inhibit PDE4 phosphodiesterase activity.
Compounds of the invention were evaluated for inhibitory activity against human platelet PDE3 to ascertain PDE3 inhibition. The PDE3 assay was performed using platelet cell extract as described above in Biological Example 1 for the PDE4 assay. Platelets are known contain PDE's 2, 3 and 5. However PDE2 and preferentially utilize cGMP, so in an assay with cAMP as a substrate they are not detected. In addition, under the conditions used in this assay, rolipram is without effect and the known PDE3 inhibitor trequinsin (Calbiochem; Catalogue No. 382425) is a potent inhibitor confirming that the assay is specific for PDE3.
Human platelet cytoplasmic extracts were prepared by modified method of Keller et al. Human platelets (AllCells, LLC; Cat. No. PB027) were sonicated in supplemented lysis buffer. Supplemented lysis buffer consisted of 20 mM Tris, 1 mM EDTA, 1 mM DTT, 0.25 M sucrose, 1 mM benzamidine, 1 μg/mL Leupeptin, 1 μM Pepstatin, and 0.1 mM PMSF, pH 7.5. The platelet lysates were centrifuged at 70,000 g for 30 minutes at 4° C. The supernatants were aliquoted and stored at −80° C.
Reactions were performed in duplicate by the addition of 10 μL PDE3 lysate to 20 μL of assay mix and 20 μL of test compounds in Isoplates (Wallac) for 30 minutes, at 37° C. The final assay mixture contained: 50 mM Tris (pH 7.5), 8.3 mM MgCl2, 1.7 mM EGTA, 0.1% BSA, and [3H]cAMP (0.025 μCi) (Amersham). Assay was terminated by addition 25 μL SPA beads and 4.75 mM IBMX (non-selective PDE inhibitor) to stop the reaction. The plate was sealed, shaken for 1 minute and then allowed to settle 30 minutes and the cpm determined using a Wallac Micobeta.
Compounds of invention were dissolved in 100% DMSO and diluted such that the final DMSO concentration in the assay did not exceed 1% to avoid affecting the PDE3 activity. PDE3 enzyme was added in quantities such that less than 15% of substrate was consumed (linear assay conditions). Test compounds were assayed at 6-8 concentrations of ranging from 0.1 nM to 100 μM and IC50 values determined from the concentration curves by nonlinear regression analysis.
Compounds of invention were evaluated for inhibition specificity for phosphodiesterases 1-11. Assay procedures are similar to those described above for in vitro PDE4 inhibition with substitution of cAMP with cGMP for PDE's that hydrolyze cGMP preferentially.
Compounds of the invention were evaluated for inhibitory activity against lipopolysaccharide (LPS) induced TNF-α release from human peripheral blood mononuclear cells (PBMC's). TNF-α is one of the most harmful endogenous pro-inflammatory cytokine. Production of this cytokine has been repeatedly shown to be potently inhibited in the presence of PDE4 inhibitors in vivo and in vitro which are believed to contribute largely to the anti-inflammatory effects of these drugs, at least under acute inflammatory conditions (Draheim, R. et al., “Anti-Inflammatory Potential of the Selective Phosphodiesterase 4 Inhibitor N-(3,5-Dichloro-pyrid-4yl)-[1-(4-fluorobenzyl)-5-hydroxy-indole-3-yl]-glyoxylic Acid Amide (AWD 12-281), in Human Cell Preparations”, The Journal of Pharmacology and Experimental Therapeutics (2004), Vol. 308, No. 2: 555-563; and Billah, M. M. et al., “Pharmacology of N-(3,5-Dichloro-1-oxido-4-pyridinyl)-8-methoxy-2-(trifluoromethyl)-5-quinoline Carboxamide (SCH 351591), a Novel, Orally Active Phosphodiesterase 4 inhibitor”, The Journal of Experimental Therapeutics (2002), Vol. 302, No. 1:127-137). The most potent TNF-α producing cell populations belong to the monocyte/macrophage lineage.
There are number of publications showing inhibition of TNF-α release by known PDE4 inhibitors in whole blood as well as isolated PBMC's (Schindler, R. et al., “Correlations and Interactions in the Production of Interleukin-6 (IL-6), IL-1, and Tumor Necrosis Factor (TNF) in Human Blood Mononuclear Cells: IL-6 Suppresses IL-1 and TNF”, Blood (1990), Vol. 75, No. 1: 40-47). Studies with PDE4B knock-out mice revealed that this PDE4 subtype was essential for LPS-induced generation of TNF-α. Therefore, testing the compounds of the invention in this assay served as a convenient cellular screening. The assay validated the ability of compounds of invention to enter the cells and will show some specificity towards desired activity against PDE4B.
PBMC's were purchased from AllCells, LLC and were prepared according to the manufacturer protocol. Briefly, the frozen vial of PBMC's (AllCells, LLC Catalogue No. PB003F) was removed from cryostorage, thawed quickly in 37° C. water bath and transferred to 50 mL tube containing 300 μg of DNAse. The vial was rinsed with 1 mL of supplemented RPMI 1640 medium pre-warmed in incubator (5% CO2, 37° C.). Supplemented RPMI 1640 contained 10% FBS, 2 mM L-Glutamine, Penicillin 50 Units/Streptomycin 50 μg/mL and 10 mM HEPES. The volume was brought to 20 mL with supplemented medium slowly. Cells were centrifuged twice for 15 minutes at 2400 rpm at ambient temperature. PBMC's were counted, diluted to approximately 0.7×106/mL and 150 μL aliquots were dispensed to each well of a tissue culture treated 96-well plate for a final cell density of 1×105/well. As defined by supplier, such preparation contained 12% of monocytes (CD14+). Viability of the cells in each experiment exceeded 90%. Plates were placed in incubator (5% CO2, 37° C.) for 0.5-1 h to allow monocytes to attach.
Compounds of invention were diluted in V-bottom 96-well plate. Test compounds were first diluted in 100% DMSO, followed by dilutions in enriched RPMI medium to give a final assay DMSO concentration of 0.03%. Plates containing PBMC's were removed from incubator and 40 μL of test compounds were added and the PBMC's pre-incubated with the test compounds for 1 h.
After 1 h pre-incubation with the tested compounds, 10 μL of LPS was added and the plates incubated (5% CO2, 37° C.) for 18 hours. Final LPS concentration in the assay was 1 ng/mL. Plates were then centrifuged at 2200 rpm (˜786 g) for 10 minutes at ambient temperature. 50 μL cell culture supernatants were carefully removed and frozen immediately at −80° C.
Human TNF-α content was determined by ELISA. Cell supernatants were diluted 15 times with appropriate ELISA diluent. Results were obtained either with BD Human TNF-α ELISA (BD) or BioSource ELISA (BioSource). In general, the ELISA assay procedure involved coating Immulon 4 HBX strips or plates with capture antibody (anti-human TNF), washing with PBST (PBS with 0.05% Tween-20) and blocking with 1% BSA or 10% FBS. After another washing, 2 hour incubation with the diluted cell supernatants or standard (recombinant human TNF) was done. Subsequent washing was followed by incubation with detection antibody (biotinylated anti-human TNF) and enzyme reagent (streptavidin-horseradish peroxidase conjugate). After final PBST washing, enzyme substrate (tetramethylbenzidine/hydrogen peroxide) was added. Reaction was stopped by addition of stop solution (2 N sulphuric acid). Read out was obtained by absorbance measurement at 450 nm (reference filter 650 nm) with Multiscan Spectrum plate reader.
Test compounds were assayed at 6-8 concentrations of ranging from 1 nM to 10 μM and IC50 values determined from the concentration curves by nonlinear regression analysis. The inhibition of LPS induced TNF-α release from human PBMC's thereof served as a convenient cellular assay to evaluate PDE4 inhibitors.
In order to demonstrate the ability of compounds of the present invention to elevate cAMP in intact cells, transfection of cells with a plasmid construct containing a cAMP response element (CRE) in a promoter driving the expression of a luciferase reporter gene (Stratagene; Path Detect™: Catalogue No. 219076) was used to allow sensitive monitoring of intracellular cAMP levels through detection of light output in a luminometer. Pharmacological treatment of transfected cells with a compound providing a combination of PDE inhibitor and adenylyl cyclase agonist (receptor or intracellular activator) resulted in elevated intracellular cAMP levels detectable from increased light output. PDE4 has been shown to be the predominant cyclic nucleotide phosphodiesterase activity in U937 cells, and therefore this cell type transfected with the CRE-luciferase construct served as a convenient cellular screening assay for compounds with PDE4 inhibitory activity. Compounds of the present invention were thereby shown to provide potentiated luciferase expression in U937 cells treated with the adenylyl cyclase activator forskolin.
U937 cells were maintained in RPMI medium containing 10% FCS and 2 mM glutamate. U937 cells were transiently transfected as described in Biotechniques (1994), Vol. 17(6):1058. Briefly, cells were grown in medium containing serum to a density of 5×106 cells/mL and then resuspended in media containing serum at a density of approximately 1×107 cells/mL. 400 μL of cells were transferred into the electroporation cuvette containing 10 μg of the reporter vector (pCRE-luc) in a volume of 40 μL H2O. Reporter vector DNA was prepared from DH5 α E. coli using the DNA endonuclease free kit (Qiagen) as per manufacturers instructions. U937 cells were electroporated at ambient temperature using a BIORAD electroporator. Capacitance was set to 1050 μμF and voltage was 280V. The time constant was noted after each electroporation. Cells were then diluted in 4 mL of media and serum and 200 μμL of cells were plated per well. Cells were allowed to recover for 16-18 hours. Cells were then treated with a test compound or vehicle in the presence or absence of 10 μM forskolin for 4 hours at 37° C.
The luciferase assay was performed as per manufacturer's instructions (Tropix).
Briefly, cells were centrifuged for 4 minutes at 1200 rpm and media supernatant was removed. Cell pellets were lysed in 15 μL Lysis buffer (Tropix). Luciferase assay was performed using 10 μμL of cell lysate with 10 μL of buffer A and 25 μL buffer B. Luciferase activity was obtained using a luminometer with a 5 second delay followed by a read time of 10 seconds.
None of the test compounds in the absence of stimuli induced significant luciferase activity indicating a low basal adenylyl cyclase activity in these cells. This result demonstrated that the compounds tested were capable of elevating cAMP levels in a cell line predominantly expressing PDE4 consistent with the observations in the enzymatic assays.
Delayed type hypersensitivity models are T cell dependent responses. The type of chemical hapten used can bias the T cell response towards a predominantly TH1 or TH2 polarization. Oxazolone and di-nitro-chloro-benzene (DNCB) induce a TH1 dominant immune response.
Mice are sensitized on day 0 by epicutaneous application of 100 μL 3% oxazolone solution in 95% ethanol on the shaved abdomen. This procedure is repeated on day 1. Six days after sensitization (i.e., on day 5), mice are challenged by topically painting 25 μL 0.8% oxazolone dissolved in 95% ethanol on both sides of the right ears and 25 μL of 95% ethanol on the left ears. On day 6 (24 hours after challenge), mice are sacrificed, both ears are removed and a standard disc of tissue is harvested immediately from each ear using a cork borer. Care is taken to sample the tissues from the same ear area. The weight of the ear disc tissues is immediately measured. Test compounds are administered orally at a dose of 5 mg/kg once daily for 7 days (from day 0 to day 6) with the last dose 2 hours prior to sacrifice.
Alternatively, mice are sensitized on day 0 by epicutaneous application of 50 μL 1% di-nitrochlorobenzene (DNCB) solution in 4: ratio of acetone:olive oil on the shaved abdomen. This procedure is repeated on day 5. Starting eleven days after the initial sensitization, mice are challenged 3 times (on days 10, 11, and 12) by topically painting 25 μL 0.5% DNCB dissolved in a 4:1 ratio of acetone:olive oil on both sides of the right ears and 25 μL of vehicle on the left ears. Twenty-four hours after challenge, mice are sacrificed as described above. Test compounds are administered orally at a dose of 10 mg/kg once daily for 5 days (from day 8 to day 12) with the last dose 2 hours prior to challenge.
Ear edema is expressed as increase in ear weight, and calculated by subtracting the left ear weight (challenged with vehicle) from that of right ear (challenged with chemical hapten). The percentage inhibition of the ear edema by drugs is calculated using following equation: 100-((drug edema/mean control edema)*100).
Compounds of the invention may, inhibit oxazolone and DNCB induced dermal inflammation at doses of less than 20 mg/kg.
Mice are sensitized on day 0 by epicutaneous application of 50 μL 0.5% fluorescein isothiocyanate (FITC) solution in 1:1 acetone and dibutyl phthalate on the shaved abdomen. This procedure is repeated on day 7. Fourteen days after sensitization (i.e., on day 13), mice are challenged by topically painting 25 μL 0.5% FITC dissolved in 1:1 acetone and dibutyl phthalate on both sides of the right ears and 25 μL 1:1 acetone and dibutyl phthalate solution on the left ears. On day 14 (24 hours after challenge), mice are sacrificed, both ears are removed and a standard disc of tissue is harvested immediately from each ear using a cork borer. Care is taken to sample the tissues from the same ear area. The weight of the ear disc tissues is immediately measured. Test compounds (5-10 mg/kg) or vehicle is administered orally once daily for 3 days (from day 11 to day 13) 2 hours prior to challenge.
Ear edema is expressed as increase in ear weight, and calculated by subtracting the left ear weight (challenged with vehicle) from that of right ear (challenged with FITC). The percentage inhibition of the ear edema by drugs is calculated using the following equation: 100-((drug edema/mean control edema)*100). Compounds of the invention may inhibit FITC induced dermal inflammation at doses of less than 20 mg/kg.
A number of mice are uniquely identified by placing a mark with an indelible marker on their tail. Mice are dosed orally with 15 mg/kg test compound in 100 μL of 45% β-cyclodextrin in saline. Mice are briefly anaesthesized with 2% halothane, and 2 μg of phorbol 12-myristate 13-acetate (PMA) in 25 μl of acetone is applied to the inner and outer sides of the left ear of the mouse. Acetone is applied to the right ear of the mouse in the same manner to serve as a vehicle control. Control animals receive the same treatment but without any test compound. After 3 hours, mice are sacrificed by cervical dislocation, and a standard sized biopsy is excised from the ears and weighed to the nearest 1/10th of a mg. Data are analyzed by taking the difference of each left ear from the right ear, and then calculating the % inhibition of edema by (((mean Rx/mean irritant))×100)−100.
Compounds of the invention may inhibit PMA induced dermal edema at doses of less than 20 mg/kg.
The collagen-induced arthritis (CIA) model in mice is a suitable model for evaluating potential drugs active in human rheumatoid arthritis. It shares many of the molecular, cellular and histopathological changes identified as hallmarks of the human disease; these include (a) pronounced proliferation of cells comprising the joint synovial membrane, (b) formation of an invasive pannus-like tissue, (c) macrophage, granulocyte and lymphocytic infiltration and (d) destruction of bone and cartilage. Like rheumatoid arthritis, animals with CIA exhibit elevated serum levels of immunoglobulin complexes such as rheumatoid factor (RF) and anti-collagen antibodies and inflammatory cytokines in the synovium such as tumour necrosis factor (TNF-α). In addition, involvement of MHC class II-restricted T-helper cell activation/clonal expansion in the synovium has been demonstrated. Radiographs of affected joints often show erosive changes similar to those seen in human RA and the progressive arthritis often results in an RA-like joint deformity and dysfunction. In addition, many compounds which reduce the symptoms of human disease such as anti-TNF biologics, corticosteroids and DMARDS are efficacious in this animal model. The development/progression of disease in the CIA model occurs in both an immune (early) and inflammatory phase thus allowing the assessment of a wide range of drugs with diverse pharmacological modes of action.
Male DBA/1J mice (7-8 weeks of age) are immunized through a subcutaneous injection of 0.1 mL of a collagen-adjuvant emulsion (0.1 mg chick type II collagen in complete Freund's adjuvant) at the base of the tail. Mice are then randomly assigned to treatment or control groups. After three weeks the animals are boosted with a second injection of chick type II collagen emulsified at 1.0 mg/mL in incomplete Freund's adjuvant. This second injection is required for reproducible induction of disease. In control animals, clinical signs of arthritis manifested as erythema and edema of the paws and tarsal/metatarsal joints usually appear within 1-2 weeks following the second immunization. Compounds are evaluated for their ability to delay the onset of or reduce the development of arthritis (prophylactic regime). Compounds are administered twice daily beginning on the day of the second collagen injection. The mice continue to receive test article until the last animal in the vehicle control group reached the seventh day of having established disease (approximately 25 days).
The development of clinical arthritis (disease progression) is monitored daily after the second collagen injection. All four limbs are clinically evaluated by a trained observer unfamiliar (blinded) with the treatment group identity, and scored on a scale of 0-4 for disease severity (redness and swelling) according to the following criteria.
Inflammation is defined as any redness or swelling (enlargement) of any part of any paw. Established disease was defined as a qualitative score of paw inflammation of 2 or greater, that persists for at least 24 hours. In addition, paw widths for all four limbs were measured by a blinded observer daily using precision, constant tension calipers.
At the end of the study each animal is euthanized by an overdose of halothane anesthesia. Joints both distal to the knee and including the knee are dissected and analyzed by histology. Limb joints are fixed in 10% formalin buffer and decalcified in 10% formic acid for 48 hours, then processed for paraffin embedding. Serial sections (5-7 micrometer thick) are stained with haematoxylin and eosin (H & E). Histopathological alterations of the tarsal and metatarsal joints are graded “blind” by a certified pathologist and a score assigned based on a ranking system. ANOVA and appropriate post-hoc test will be used to determine if arthritis scores from test article treated animals will be significantly lower than those of the vehicle treated animals.
Compounds of the invention may inhibit clinical signs of CIA-induced arthritis at doses of less than 20 mg/kg.
This model is used to investigate the effect of novel compounds on cartilage degradation induced by the natural inflammatory response created by implantation of a foreign body. Activity in this model may be indicative of activity in arthritis.
Zyphoid sternum cartilage is excised from CO2 terminated rats, washed in Hibitane, and rinsed in sterile, phosphate buffered saline. A 4 cm diameter disc is removed from the sternum with a #4 stainless steel leather hole punch, and cut in half. Each half is weighed and wrapped in pre-weighed, moist, sterile cotton before implantation. A piece of cotton wrapped cartilage is implanted subcutaneously into each dorsolateral surface of anaesthetized female CD/1 mice (aged 6-8 weeks) via a 1 cm incision along the dorsal midline (Day 0). Mice are administered test articles by oral administration on days 3 to 17. On day 18, mice are sacrificed, the cotton and cartilage removed, and the cartilage separated from the cotton. Both the cartilage and the cotton are weighed, and differences between pre and post implant weights are calculated. The cotton is rinsed in 1 mL of buffer, and cytospins are prepared and stained for differentiation and enumeration of cell types. In addition, the resuspended lavage fluid is analyzed for absolute cell numbers and cell differentials by the CellDyn 3700SC hematology analyzer (Abbott Laboratories Inc.).
The cartilage is digested overnight in a papain and cysteine hydrochloric acid solution at 65° C. and glucosaminoglycan content remaining in the cartilage is assayed by spectrophotometrically and calculated as % GAG/mg of cartilage degraded (normalized to pre implant cartilage weight).
Compounds of the invention may inhibit cartilage degradation at doses of less than 20 mg/kg.
This model is used to investigate the effect of novel compounds on joint inflammation induced by LPS. Joint inflammation occurs in the joints of patients with rheumatoid arthritis. Activity in this model may be indicative of activity in arthritis.
Balb/C mice will be injected directly into left hind knee joint with 3 ng of LPS (6 μl of stock) using an Hamilton syringe (H80401) adapted to a 30 G needle. A 9 mm long spacer made of PE10 tubing will be placed on needle to insure LPS was injected to the same depth for each animal. Care will be taken to ensure no fluid is drawn back after each injection. The same volume of saline (6 μl) will be injected into the right hind knee, as a control, using a separate Hamilton syringe. Eighteen hours after challenge, animals will be anesthetized with 5% isoflurane and euthanized by cardiac puncture. The hind limbs will be dissected free from attached muscles and removed. The synovial cavity of each leg will be exposed by pulling the patellar tendon towards the distal end of the leg, and will be washed with 3 ml of ice-cold EDTA (10 mM)-PBS buffer. The washout solution will be centrifuged at 1200 rpm for 3 min. Supernatant will be removed and the cell pellet will be resuspended in 0.5 ml cold PBS/EDTA. Total cell counts and differentials in the synovial washout will be counted using a Cell Dyne hematology analyzer and cytospin preparations.
Data will be plotted as mean±SEM. One-way ANOVA followed by student Newman-Kuels all-pairwise or Dunnett's post-hoc test will be used for comparison of multiple means. P<0.05 will be considered statistically significant. Compounds of the invention may inhibit joint inflammation at doses of less than 20 mg/kg.
Rats are administered drug (1-20 mg/kg) or vehicle orally once (0-24 hours) prior to challenge. Rats are challenged with either saline or LPS dissolved in saline (2 mg/kg) via intra-tracheal installation. Animals are sacrificed via intra-peritoneal sodium pentobarbitol overdose 3 hours post challenge, and the lungs lavaged with 14 mL of phosphate buffered saline (PBS). The lung lavage fluid is centrifuged at 300 g for 3 min, and the supernatant removed. The pellet is resuspended in 1-3 mL of PBS at 4° C. depending on pellet size and numbers of total leukocytes. A volume of the final cell suspension, containing approximately 240,000 cells, is added to an appropriate volume of PBS at 4° C. to give a final volume of 220 μL and a final concentration of 1×106 cells/mL (final Cytospin suspension). A 100 μL sample (100,000 cells) is loaded onto a cytospin centrifuge and spun for 4 min at 55 g. Two slides are prepared per lavage sample, and are fixed and stained in DifQuik. In addition, the resuspended lavage fluid is analyzed for absolute cell numbers and cell differentials by the CellDyn 3700SC hematology analyzer (Abbott Laboratories Inc.). This model could be adapted to assess effect of selected compounds in a mouse model of LPS-induced lung inflammation.
Compounds of the invention may inhibit LPS induced lung inflammation at doses of less than 20 mg/kg.
The ability of a compound to inhibit the allergen-induced accumulation of inflammatory cells such as eosinophils and neutrophils in the lavage fluid obtained from sensitized animals is indicative of that compound's anti-asthma activity. In particular, this model system is useful in the evaluation of the effects of a test compound in the treatment of the late phase response of asthma, when lung inflammation and the second phase of bronchoconstriction is apparent, and in allergy, especially where it affects the respiratory system. The test is conducted as follows.
Male Brown Norway rats are sensitized to ovalbumin by single intraperitoneal injection of 1 mg ovalbumin adsorbed to 100 mg Al(OH)3 (alum) in 1 mL sterile saline (saline control rats receive only sterile saline) on day 1, and allowed to sensitize until day 21. Test compounds are given orally q.d. for three days prior to challenge (days 19, 20, 21), and one day post challenge (day 22), with the third dose given 2 hours before challenge, and the fourth day dose given 24 hours after challenge (volume=300 μl/dose). Rats are challenged with 5% ovalbumin in saline generated using a Devillbis nebulizer for 5 min on day 21.
Forty-eight hours after challenge, animals are sacrificed with an overdose of intraperitoneally-delivered sodium pentobarbitol and the lungs are lavaged with cold 2×7 mL phosphate buffered saline. The recovered lavage fluid is placed on ice. The bronchoalveolar lavage fluid is centrifuged and the supernatant removed. The pellet is resuspended in phosphate buffered saline at 4° C. Cytospins are prepared and stained for differentiation and enumeration of cell types. This model could be adapted to assess effect of selected compounds in a mouse model of allergen-induced lung inflammation.
Compounds of the invention may inhibit allergen induced lung inflammation at doses of less than 20 mg/kg.
The Buxco murine airway hyper-responsiveness (AHR) model has been well characterized by numerous investigators, and mimics the severe airway constriction in response to aerosol challenges that sensitized animals exhibit compared to unsensitized animals. The Buxco system uses a technique called whole body plethysmography, in which breathing-induced changes in chamber pressure are quantified using the correlation between increased airway resistance and increased expiratory time/breathing pause to calculate the degree of airway constriction (Penh). Following allergen sensitization and inhalation challenge of the airway, the Penh will increase compared to sham sensitized, sham challenged animals. Thus the effectiveness of a potential anti-inflammatory agent can be determined by examining its impact on ovalbumin induced AHR.
Female Balb/c mice are sensitized on day 1 and 14 by i.p. injection of 100 μL sterile saline containing 20 μg ovalbumin and 2.25 mg Al(OH)3. Sham sensitized mice receive 100 μL sterile saline alone. Test compounds (5 mg/kg) are administered by oral gavage on five consecutive days, two days before challenge (days 26 and 27) and on the three days of ovalbumin challenge (days 28, 29 and 30, 2 hours before challenge). Mice are challenged with aerosolized ovalbumin (5% in saline) for 20 min on days 28, 29 and 30. On day 31, mice are placed in the whole body plethysmography chambers of the Buxco system and airway reactivity to aerosolized PBS and methacholine (MCh; 0.78, 1.56, 3.125, 6.25, 12.5, 25 mg/mL) challenge is measured as Penh.
Compounds of the invention may inhibit allergen induced airway hyper-reactivity at doses of less than 20 mg/kg.
Inflammatory bowel disease (IBD) is an umbrella term for presently incurable, chronic, fluctuating inflammatory diseases of the gastrointestinal tract including Crohn's disease and ulcerative colitis.
The dextran sodium sulphate (DSS) induced colitis model in mice has been shown to mimic the nature of the human disease, produce lesions that are histopathologically similar to those in humans with similar clinical pathology to that of human disease including, necrosis, formation of ulcers, granulocytic infiltration, edema of the bowel, diarrhea and adhesions with many drugs used to treat human IBD showing activity in the DSS model.
Colitis is induced by oral administration of DSS (in drinking water) (2.5-3% DSS) to groups of 8 female, CD-1 or C57BL/6 mice weighing 15-25 g. Body weight, clinical signs, diarrhea, colonic myeloperoxidase levels and histopathology for ulceration are viable and relevant endpoints.
Compounds of the invention may reduce the effects of DSS on the above endpoints in rats at doses of less than 20 mg/kg.
The trinitrobenzenesulfonic acid (TNBS) induced colitis model in rat (Morris et al., Gastroenterology 96:795-803, 1989; Kim, H.-S. and Berstad, A., Scandinavian Journal of Gastroenterology 27:529-537, 1992; Ward, Lancet ii:903-905, 1977; and Shorter et al., Am. J. Dig. Dis. 17:1024-1032, 1972)) has been shown to mimic the relapsing/remitting nature of the human disease, produce lesions that are histopathologically similar to those in humans with similar clinical pathology to that of human disease including, necrosis, formation of ulcers, granulocytic infiltration, edema of the bowel, diarrhea and adhesions with many drugs used to treat human IBD showing activity in the TNBS model.
Colitis is induced by intracolonic instillation of the hapten TNBS (60 mg/mL) in 0.5 mL of 50% ethanol to groups of 8 male, Wistar rats weighing 175-225 g. Body weight, diarrhea, colonic myeloperoxidase levels and histopathology for ulceration are viable and relevant endpoints.
Compounds of the invention may reduce the effects of TNBS on the above endpoints in rats at doses of less than 20 mg/kg.
Several behavioral models of learning and memory exist. They revolve around the ability of animals to recall a previous exposure to an object, environment or stimulus. The behavior of an animal that recalls such an exposure is usually different from the behavior of an animal that is naïve to that exposure. One example of such a model is an object recognition model using a mouse which has been engineered to be heterozygous for CREB binding protein (CBP). A similar heterozygous trait occurs in humans with Rubinstein Taybi Syndrome. One characteristic of this syndrome and these mice is the ability to form short-term memory, but no long term memory. Also CBP function is reduced, but not abolished. Increasing cAMP levels with a PDE4 inhibitor is hypothesized to enhance long term memory generation via and enhancement of CBP activity.
Object recognition in mice can be easily assessed since mice are naturally inquisitive of new objects. Mice are firstly exposed to two identical but novel objects and allowed to investigate them for 15 minutes. The objects are removed and twenty four hours later the mice are re-introduced to one such object and a second completely novel object. If the mouse has long term memory capacity it will preferentially ignore the training object and spend more time investigating the novel object. If there is no long term memory the mice will spend equal time investigating the two objects. The CBP heterozygous mice behave in this manner and have no recall of the training object.
Compounds of the invention may improve object recognition and hence learning and memory in such a mouse object recognition model at doses of less than 20 mg/kg.
Several behavioral models of learning and memory exist. They revolve around the ability of animals to recall a previous exposure to an object, environment or stimulus. The behavior of an animal that recalls such an exposure is usually different from the behavior of an animal that is naïve to that exposure. One example of such a model is a mouse fear conditioning model. Mice are trained to recognize an environment that will supply them with an unwanted stimulus, such as a mild foot shock, following an audible tone. In mice that have formed long term memory of the training environment exposure to the same environment and audible tone three days later produces a fear “freeze” response.
Compounds of the invention may improve long term memory consolidation and enhance the “freeze” response of normal or aged mice in this fear conditioning model at doses of less than 20 mg/kg.
In humans, depression is commonly characterized by a feeling of helplessness and loss of interests and energy. Clinical depression is hypothesized to result from an imbalance of signals in the brain. Classical anti-depressants, such as the tricyclic antidepressant, desipramine, increase the levels of one of these signals and help to restore signal balance. PDE4 is an enzyme responsible for turning off this signal. Therefore treatment with PDE4 inhibitors is hypothesized to enhance the brain signal as is achieved with the classical antidepressants, albeit via a novel mechanism.
The mouse forced swim test exposes mice to a setting that induces helplessness and frustration, capturing the essence of clinical depression. The forced swim test is one of the most widely used pharmacological method of assessing depression. The test involves placing a mouse in a cylinder of water and then monitoring the time spent swimming versus the time spent floating immobile (helpless time). Immobility indicates a state of despair where the animal has realized that escape is impossible and resigns itself to floating helplessly.
Antidepressant compounds, such as desipramine, that have demonstrated therapeutic effect in humans have also decreased the time of immobility in mice in the forced swim test.
Compounds of the invention may reduce immobility time of mice in the forced swim test at doses of less than 20 mg/kg.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/GB08/00855 | 3/12/2008 | WO | 00 | 3/16/2010 |
| Number | Date | Country | |
|---|---|---|---|
| 60906802 | Mar 2007 | US |
