Compositions useful as inhibitors of protein kinases

The present invention provides compounds of formula I:or a pharmaceutically acceptable derivative thereof, wherein A, B, Q, R1, and R2 are as described in the specification. These compounds are inhibitors of protein kinase, particularly inhibitors of AKT or PDK1 kinase, mammalian protein kinases involved in proliferative and neurodegenerative disorders. The invention also provides pharmaceutical compositions comprising the compounds of the invention, processes for preparing the compounds, and methods of utilizing those compositions in the treatment of various disorders.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds useful as inhibitors of protein kinases. The invention also provides pharmaceutically acceptable compositions comprising the compounds of the invention and methods of using the compositions in the treatment of various disorders.

BACKGROUND OF THE INVENTION

The search for new therapeutic agents has been greatly aided in recent years by better understanding of the structure of enzymes and other biomolecules associated with target diseases. One important class of enzymes that has been the subject of extensive study is the protein kinases.

Protein kinases mediate intracellular signal transduction. They do this by effecting a phosphoryl transfer from a nucleoside triphosphate to a protein acceptor that is involved in a signaling pathway. There are a number of kinases and pathways through which extracellular and other stimuli cause a variety of cellular responses to occur inside the cell. Examples of such stimuli include environmental and chemical stress signals (e.g. osmotic shock, heat shock, ultraviolet radiation, bacterial endotoxin, H2O2), cytokines (e.g. interleukin-1 (IL-1) and tumor necrosis factor α (TNF-α)), and growth factors (e.g. granulocyte macrophage-colony-stimulating factor (GM-CSF), and fibroblast growth factor (FGF). An extracellular stimulus may effect one or more cellular responses related to cell growth, migration, differentiation, secretion of hormones, activation of transcription factors, muscle contraction, glucose metabolism, control of protein synthesis and regulation of cell cycle.

Many diseases are associated with abnormal cellular responses triggered by protein kinase-mediated events. These diseases include autoimmune diseases, inflammatory diseases, neurological and neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, Alzheimer's disease or hormone-related diseases. Accordingly, there has been a substantial effort in medicinal chemistry to find protein kinase inhibitors that are effective as therapeutic agents. A challenge has been to find protein kinase inhibitors that act in a selective manner. Since there are numerable protein kinases that are involved in a variety of cellular responses, non-selective inhibitors may lead to unwanted side effects.

AKT (also known as PKB or Rac-PK beta), a serine/threonine protein kinase, has been shown to be overexpressed in several types of cancer and is a mediator of normal cell functions [(Khwaja, A.,Nature,401, pp. 33-34, 1999); (Yuan, Z. Q., et al.,Oncogene,19, pp. 2324-2330, 2000); (Namikawa, K., et al.,J. Neurosci.,20, pp. 2875-2886, 2000)]. AKT comprises an N-terminal pleckstrin homology (PH) domain, a kinase domain and a C-terminal “tail” region. Three isoforms of human AKT kinase (AKT-1, -2 and -3) have been reported so far [(Cheng, J. Q.,Proc. Natl. Acad. Sci. USA,89, pp. 9267-9271, 1992); (Brodbeck, D. et al.,J. Biol. Chem.274, pp. 9133-9136, 1999)]. The PH domain binds 3-phosphoinositides, which are synthesized by phosphatidyl inositol 3-kinase (PI3K) upon stimulation by growth factors such as platelet derived growth factor (PDGF), nerve growth factor (NGF) and insulin-like growth factor (IGF-1) [(Kulik et al.,Mol. Cell. Biol.,17, pp. 1595-1606, 1997); (Hemmings, B. A.,Science,275, pp. 628-630, 1997)]. Lipid binding to the PH domain promotes translocation of AKT to the plasma membrane and facilitates phosphorylation by another PH-domain-containing protein kinases, PDK1 at Thr308, Thr309, and Thr305 for the AKT isoforms 1, 2 and 3, respectively. A second, as of yet unknown, kinase is required for the phosphorylation of Ser473, Ser474 or Ser472 in the C-terminal tails of AKT-1, -2 and -3 respectively, in order to yield a fully activated AKT enzyme.

Manifestations of altered AKT regulation appear in both injury and disease, the most important role being in cancer. The first account of AKT was in association with human ovarian carcinomas where expression of AKT was found to be amplified in 15% of cases (Cheng, J. Q. et al.,Proc. Natl. Acad. Sci. U.S.A.,89, pp. 9267-9271, 1992). It has also been found to be overexpressed in 12% of pancreatic cancers (Cheng, J. Q. et al.,Proc. Natl. Acad. Sci. U.S.A.,93, pp. 3636-3641, 1996). It was demonstrated that AKT-2 was over-expressed in 12% of ovarian carcinomas and that amplification of AKT was especially frequent in 50% of undifferentiated tumours, showing that AKT is also associated with tumour aggressiveness (Bellacosa, et al.,Int. J. Cancer,64, pp. 280-285, 1995):

The 3-phosphoinositide-dependent protein kinase-1 (PDK1) plays a key role in regulating the activity of a number of kinases belonging to the AGC subfamily of protein kinases (Alessi, D. et al.,Biochem. Soc. Trans,29, pp. 1, 2001). These include isoforms of protein kinase B (PKB, also known as AKT), p70 ribosomal S6 kinase (S6K) (Avruch, J. et al.,prog. Mol. Subcell. Biol.,2001, 26, pp. 115, 2001), and p90 ribosomal S6 kinase (Frödin, M. et al.,EMBO J.,19, pp. 2924-2934, 2000). PDK1 mediated signaling is activated in response to insulin and growth factors and as a consequence of attachment of the cell to the extracellular matrix (integrin signaling). Once activated these enzymes mediate many diverse cellular events by phosphorylating key regulatory proteins that play important roles controlling processes such as cell survival, growth, proliferation and glucose regulation [(Lawlor, M. A. et al.,J. Cell Sci.,114, pp. 2903-2910, 2001), (Lawlor, M. A. et al.,EMBO J.,21, pp. 3728-3738, 2002)]. PDK1 is a 556 amino acid protein, with an N-terminal catalytic domain and a C-terminal pleckstrin homology (PH) domain, which activates its substrates by phosphorylating these kinases at their activation loop (Belham, C. et al.,Curr. Biol.,9, pp. R93-R96, 1999). Many human cancers including prostate and NSCL have elevated PDK1 signaling pathway function resulting from a number of distinct genetic events such as PTEN mutations or over-expression of certain key regulatory proteins [(Graff, J. R.,Expert Opin. Ther. Targets,6, pp. 103-113, 2002), (Brognard, J., et al.,Cancer Res.,61, pp. 3986-3997, 2001)]. Inhibition of PDK1 as a mechanism to treat cancer was demonstrated by transfection of a PTEN negative human cancer cell line (U87MG) with antisense oligonucleotides directed against PDK1. The resulting decrease in PDK1 protein levels led to a reduction in cellular proliferation and survival (Flynn, P., et al.,Curr. Biol.,10, pp. 1439-1442, 2000). Consequently the design of ATP binding site inhibitors of PDK1 offers, amongst other treatments, an attractive target for cancer chemotherapy.

The diverse range of cancer cell genotypes has been attributed to the manifestation of the following six essential alterations in cell physiology: self-sufficiency in growth signaling, evasion of apoptosis, insensitivity to growth-inhibitory signaling, limitless replicative potential, sustained angiogenesis, and tissue invasion leading to metastasis (Hanahan, D. et al.,Cell,100, pp. 57-70, 2000). PDK1 is a critical mediator of the PI3K signalling pathway, which regulates a multitude of cellular function including growth, proliferation and survival. Consequently inhibition of this pathway could affect four or more of the six defining requirements for cancer progression, as such it is anticipated that a PDK1 inhibitor will have an effect on the growth of a very wide range of human cancers.

Specifically, increased levels of PI3K pathway activity has been directly associated with the development of a number of human caners, progression to an aggressive refractory state (acquired resistance to chemotherapies) and poor prognosis. This increased activity has been attributed to a series of key events including decreased activity of negative pathway regulators such as the phosphatase PTEN, activating mutations of positive pathway regulators such as Ras, and overexpression of components of the pathway itself such as PKB, examples include: brain (gliomas), breast, colon, head and neck, kidney, lung, liver, melanoma, ovarian, pancreatic, prostate, sarcoma, thyroid [(Teng, D. H. et al.,Cancer Res.,57, pp. 5221-5225, 1997), (Brognard, J. et al.,Cancer Res.,61, pp. 3986-3997, 2001), (Cheng, J. Q. et al.,Proc. Natl. Acad. Sci.,93, pp. 3636-3641, 1996),Int. J. Cancer,64, pp. 280, 1995), (Graff, J. R.,Expert Opin. Ther. Targets,6, pp. 103-113, 2002),Am. J. Pathol.,159, pp. 431, 2001)].

Accordingly, there is a great need to develop inhibitors of AKT and PDK1 protein kinases that are useful in treating various diseases or conditions associated with AKT and PDK1 activation, particularly given the inadequate treatments currently available for the majority of these disorders.

SUMMARY OF THE INVENTION

This invention provides compounds having the formula I:

These compounds, and pharmaceutically acceptable compositions thereof, are useful for treating or lessening the severity of a variety of disorders, including proliferative disorders and neurological disorders.

DESCRIPTION OF THE INVENTION

The present invention relates to a compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein:A is —CH2— or —CH2C(Ra)(Rb)—, wherein:Raand Rbare independently hydrogen, an optionally substituted C1-6aliphatic group, or halogen, or Raand Rbare taken together to form a 3-6 membered saturated or partially unsaturated ring having 0-2 heteroatoms independently selected from nitrogen, oxygen or sulfur;B is A is —CH2— or —CH2C(Rc)(Rd)—, wherein:Rcand Rdare independently hydrogen, C1-4aliphatic, or halogen, or Rcand Rdare taken together to form a cyclopropyl ring;R1is T—Ar;each T is independently selected from a valence bond or a C1-6wherein up to two methylene units of T are optionally, and independently, replaced by —O—, —N(R)—, —S—, —N(R)C(O)—, —C(O)N(R)—, —C(O)—, or —SO2—;each R is independently selected from hydrogen or an optionally substituted C1-6aliphatic group, or:two R groups on the same nitrogen, taken together with the nitrogen atom attached thereto, form a 5-7 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;Q is a valence bond or a C1-6alkylidene chain, wherein up to two methylene units of Q are optionally, and independently, replaced by —O—, —N(R)—, —S—, —N(R)C(O)—, —C(O)N(R)—, —C(O)—, or —SO2—;R2is selected from Ar, R3, or C(R)(Ar)R3, wherein:R and R3optionally form a 5-7 membered saturated or partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each Ar is independently an optionally substituted ring selected from a 5-7 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-10 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each R3is independently selected from R′, Ar1, W—OR5, W—OC(O)R5, W—CONHR5, W—OC(O)NHR5, W—SR5, W—N(R4)2, N(R)(W—Ar), N(R)C(O)W—N(R4)2, or N(R)W—N(R4)2, wherein:each W is independently a valence bond or a C1-6alkylidene chain;R′ is an optionally substituted C1-6aliphatic group;each Ar1is independently selected from an optionally substituted 5-7 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each R4is independently selected from R, COR5, CO2R5, CON(R5)2, SO2R5, SO2N(R5)2, or Ar1; andeach R5is independently selected from R or Ar;provided that:when one of A or B is —CH2— and the other of A or B is —CH2CH2—, R1is T—Ar, T is a valence bond, Ar is a 5-7 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and Q is a C1-6alkylidene chain wherein the methylene unit attached to the nitrogen atom is replaced by C(O), then R2is other than optionally substituted phenyl; andwhen T is —NH—, —NHC(O)—, or —NHC(O)N(R)—, then R2is W—C(R)(W—Ar)R3.

As used herein, the following definitions shall apply unless otherwise indicated. The phrase “optionally substituted” is used interchangeably with the phrase “substituted or unsubstituted.” Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group, and each substitution is independent of the other.

The term “aliphatic” or “aliphatic group” as used herein means a straight-chain or branched C1-C12hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic C3-C8hydrocarbon or bicyclic C8-C12hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “carbocycle” or “cycloalkyl”), that has a single point of attachment to the rest of the molecule wherein any individual ring in said bicyclic ring system has 3-7 members. For example, suitable aliphatic groups include, but are not limited to, linear or branched or alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

The terms “alkyl”, “alkoxy”, “hydroxyalkyl”, “alkoxyalkyl”, and “alkoxycarbonyl”, used alone or as part of a larger moiety includes both straight and branched chains containing one to twelve carbon atoms. The terms “alkenyl” and “alkynyl” used alone or as part of a larger moiety shall include both straight and branched chains containing two to twelve carbon atoms.

The terms “haloalkyl”, “haloalkenyl” and “haloalkoxy” means alkyl, alkenyl or alkoxy, as the case may be, substituted with one or more halogen atoms. The term “halogen” means F, Cl, Br, or I.

The term “heteroatom” means nitrogen, oxygen, or sulfur and includes any oxidized form of nitrogen and sulfur, and the quaternized form of any basic nitrogen. Also the term “nitrogen” includes a substitutable nitrogen of a heterocyclic ring. As an example, in a saturated or partially unsaturated ring having 0-4 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR+(as in N-substituted pyrrolidinyl).

The term “aryl” used alone or as part of a larger moiety as in “aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic, bicyclic and tricyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 7 ring members. The term “aryl” may be used interchangeably with the term “aryl ring”.

The term “heterocycle”, “heterocyclyl”, or “heterocyclic” as used herein means non-aromatic, monocyclic, bicyclic or tricyclic ring systems having five to fourteen ring members in which one or more ring members is a heteroatom, wherein each ring in the system contains 3 to 7 ring members.

The term “heteroaryl”, used alone or as part of a larger moiety as in “heteroaralkyl” or “heteroarylalkoxy”, refers to monocyclic, bicyclic and tricyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and wherein each ring in the system contains 3 to 7 ring members. The term “heteroaryl” may be used interchangeably with the term “heteroaryl ring” or the term “heteroaromatic”.

An aliphatic group or a non-aromatic heterocyclic ring may contain one or more substituents. Suitable substituents on the saturated carbon of an aliphatic group or of a non-aromatic heterocyclic ring are selected from those listed above for the unsaturated carbon of an aryl or heteroaryl group and the following: ═O, ═S, ═NNHR*, ═NN(R*)2, ═N—, ═NNHC(O)R*, ═NNHCO2(alkyl), ═NNHSO2(alkyl), or ═NR*, where each R* is independently selected from hydrogen or an optionally substituted C1-6aliphatic. Substituents on the aliphatic group of R* are selected from NH2, NH(C1-4aliphatic), N(C1-4aliphatic)2, halogen, C1-4aliphatic, OH, O—(C1-4aliphatic), NO2, CN, CO2H, CO2(C1-4aliphatic), —O(halo C1-4aliphatic), or halo C1-4aliphatic.

The term “alkylidene chain” refers to a straight or branched carbon chain that may be fully saturated or have one or more units of unsaturation and has two points of connection to the rest of the molecule. That is, alkylidene refers to an aliphatic group (alkyl, alkenyl, or alkynyl) that has two points of connection to the rest of the molecule.

The compounds of this invention are limited to those that are chemically feasible and stable. Therefore, a combination of substituents or variables in the compounds described above is permissible only if such a combination results in a stable or chemically feasible compound. A stable compound or chemically feasible compound is one in which the chemical structure is not substantially altered when kept at a temperature of 40° C. or less, in the absence of moisture or other chemically reactive conditions, for at least a week.

Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by a13C- or14C-enriched carbon are within the scope of this invention.

Compounds of this invention may exist in alternative tautomeric forms. Unless otherwise indicated, the representation of either tautomer is meant to include the other.

A preferred embodiment of this invention provides a compound wherein R2is —C(R)(Ar)R3.

In another preferred embodiment, R2is —C(R)(WAr)R3(where R is preferably, H).

In other preferred embodiments, R2is as depicted in compounds I-6, I-7, I-12, or I-101-I-197.

This invention also provides compounds wherein the T moiety is T′, wherein T′ is —N(R)—, —N(R)C(O)—, —N(R)C(O)NH—, —N(R)CH2—, or —N(R)SO2—;

According to one embodiment, the T moiety of the R1group of formula I is selected from a valence bond, or a C1-6alkylidene chain wherein up to two methylene units of T are optionally, and independently, replaced by —O—, —S—, —C(O)N(R)—, —C(O)—, or —SO2—. Examples of such groups include —CH2—, —CH2CH2—, —CH═CH—, —C≡C—, —CH2(CH3)—, —SC(O)—, —CH2C(O)—, —C(O)NH—, —OC(O)NH—, —O—, and —S—.

According to another embodiment, the T moiety of the R1group of formula I is selected from a valence bond, or a C1-6alkylidene chain wherein up to one methylene unit of T is optionally replaced by —N(R)—, —N(R)C(O)—, —N(R)C(O)N(R)—, —N(R)SO2—, or —N(R)SO2N(R)—. Examples of such groups include —NH—, —NHCH2—, —NHC(O)—, —NHC(O)NH—, —NHC(O)CH2—, and NHC(O)CH2CH2—. Further examples of such groups include —N(CH3)—, —N(CH3)CH2—, —N(CH3)C(O)—, and —N(CH3)SO2—.

Preferred T moieties of the T—Ar group of R1are selected from a valence bond, —N(R)C(O)—, —NH—, —NHCH2—, —NHSO2—, —CH2NH—, —SC(O)—, —CH2C(O)—, —C≡C—, —CH2— or —CH2CH2—. More preferred T moieties of the T—Ar group of R1are selected from —NHC(O)—, —NH—, —NHCH2—, —CH2—, —C≡C—, or —CH2CH2—. Most preferred T moieties of the T—Ar group of R1are selected from —N(R)C(O)—, —NH—, or —NHCH2—. In one embodiment, R1is —T—Ar, wherein T is —N(R)C(O)— and Ar is thienyl.

Preferred W groups of formula I are selected from a valence bond, —CH2—, or —CH2CH2—.

When the R2group of formula I is Ar, preferred Ar groups are an optionally substituted ring selected from a 5-6 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or a 9-10 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Examples of such monocyclic rings include phenyl, pyridyl, pyrimidinyl, pyridonyl, furanyl, tetrazolyl, thienyl, cyclopentyl, cyclohexyl, and cycloheptyl. Examples of such bicyclic rings include benzo[1,3]dioxolyl, indan-1-onyl, naphthyl, benzothiophenyl, 2,3-dihydro-1H-isoindolyl, indanyl, benzofuranyl, and indolyl.

When present, preferred substituents on the Ar ring of the R2group of formula I include Ro, halogen, oxo, ORo, phenyl, optionally substituted dialkylamino, haloalkyl, C(O)Ro, NHC(O)R, or SRo. Examples of such preferred substituents include chloro, bromo, fluoro, OH, OMe, NHC(O)CH3, OEt, C(O)phenyl, Ophenyl, N(CH2CH2Cl)2, N(Me)2, CF3, and SCF3. Other examples of preferred Ar groups of formula I also include those shown in Table 1 below.

Most preferably, the R3group of formula I is CH2CH2NH2. Other most preferred R3groups of formula I are CH2OH, CH2CH2OH, and CH2NH2.

Preferred rings formed by the R and R3moieties of the W—C(R)(W—Ar)R3group of R2are selected from a 5-6 membered saturated ring having 0-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Examples of such rings formed by R and R3include piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, and thiomorpholinyl.

When the R2group of formula I is W—C(R)(W—Ar)R3, preferred Ar groups of the W—C(R)(W—Ar)R3moiety are selected from an optionally substituted 5-6 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an optionally substituted 9-10 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Examples of such monocyclic rings include phenyl, pyridyl, furanyl, pyridone, and thienyl. Examples of such bicyclic rings include benzo[1,3]dioxolyl, naphthyl, indanyl, and indolyl. When present, preferred substituents on the Ar ring of the W—C(R)(W—Ar)R3group of R2include Ro, halogen, ORo, phenyl, N(Ro)2, NHC(O)Ro, or SRo. Examples of such groups include fluoro, chloro, bromo, CF3, OH, OMe, OPh, OCH2Ph, SMe, NH2, NHC(O)Me, methyl, ethyl, isopropyl, isobutyl, and cyclopropyl.

According to another embodiment, R3is —W—OR5. W, in this embodiment, is preferably a C1, C2, or C3 alkyl group (preferably a C1 or C2 alkyl). R5, in these embodiments, is preferably H thus forming a hydroxy group (or an appropriate derivative thereof).

According to yet another embodiment, R3is —W—N(R4)2. W, in this embodiment, is preferably a C1, C2, or C3 alkyl group (preferably a C1 alkyl). One or both R4groups, in these embodiments, is preferably H thus forming a secondary or tertiary amino group (or an appropriate derivative thereof).

According to another embodiment, the present invention relates to a compound of formula II:

or a pharmaceutically acceptable salt thereof, wherein:A is —CH2— or —CH2C(Ra)(Rb)—, wherein:Raand Rbare independently hydrogen, an optionally substituted C1-6aliphatic group, or halogen, or Raand Rbare taken together to form a 3-6 membered saturated or partially unsaturated ring having 0-2 heteroatoms independently selected from nitrogen, oxygen or sulfur;B is A is —CH2— or —CH2C(Rc)(Rd)—, wherein:Rcand Rdare independently hydrogen, C1-4aliphatic, or halogen, or Rcand Rdare taken together to form a cyclopropyl ring;R1is T′—Ar;T′ is —N(R′)—, —N(R′)C(O)—, —N(R′)C(O)NH—, —N(R′)CH2—, or —N(R′)SO2—;each R is independently selected from hydrogen or an optionally substituted C1-6aliphatic group, or:two R groups on the same nitrogen, taken together with the nitrogen atom attached thereto, form a 5-7 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;Q is a valence bond or a C1-6alkylidene chain, wherein up to two methylene units of Q are optionally, and independently, replaced by —O—, —N(R)—, —S—, —N(R)C(O)—, —C(O)N(R)—, —C(O)—, or —SO2—;R2is selected from Ar, R3, or C(R)(Ar)R3, wherein:R and R3optionally form a 5-7 membered saturated or partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each Ar is independently an optionally substituted ring selected from a 5-7 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-10 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each R3is independently selected from R′, Ar1, W—OR5, W—OC(O)R5, W—CONHR5, W—OC(O)NHR5, W—SR5, W—N(R4)2, N(R)(W—Ar), N(R)C(O)W—N(R4)2, or N(R)W—N(R4)2, wherein:each W is independently a valence bond or a C1-6alkylidene chain;R′ is an optionally substituted C1-6aliphatic group;each Ar1is independently selected from an optionally substituted 5-7 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each R4is independently selected from R, COR5, CO2R5, CON(R5)2, SO2R5, SO2N(R5)2, or Ar1; andeach R5is independently selected from R or Ar.

Preferred A, B, Ar, Q, and R2groups of formula II are those described above for compounds of formula I. Preferred T′ groups of formula II are selected from —N(R′)—, —N(R′)C(O)—, —N(R′)C(O)NH—, —N(R′)CH2—, or —N(R′)SO2— (wherein R′ is R). More preferred T′ groups of formula II are selected from —N(R′)C(O)—, —N(R′)—, —N(R′)CH2—, —N(R′)SO2— (wherein R′ is R.

According to another embodiment, the present invention relates to a compound of formula III:

or a pharmaceutically acceptable salt thereof, wherein:A is —CH2— or —CH2C(Ra)(Rb)—, wherein:Raand Rbare independently hydrogen, an optionally substituted C1-6aliphatic group, or halogen, or Raand Rbare taken together to form a 3-6 membered saturated or partially unsaturated ring having 0-2 heteroatoms independently selected from nitrogen, oxygen or sulfur;B is A is —CH2— or —CH2C(Rc)(Rd)—, wherein:Rcand Rdare independently hydrogen, C1-4aliphatic, or halogen, or Rcand Rdare taken together to form a cyclopropyl ring;R1is T—Ar;each T is independently selected from a valence bond or a C1-6alkylidene chain, wherein up to two methylene units of T are optionally, and independently, replaced by —O—, —N(R)—, —S—, —N(R)C(O)—, —C(O)N(R)—, —C(O)—, or —SO2—;each R is independently selected from hydrogen or an optionally substituted C1-6aliphatic group, or:two R groups on the same nitrogen, taken together with the nitrogen atom attached thereto, form a 5-7 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each Ar is independently an optionally substituted ring selected from a 5-7 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-10 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each R3is independently selected from R′, Ar1, W—OR5, W—OC(O)R5, W—CONHR5, W—OC(O)NHR5, W—SR5, W—N(R4)2, N(R)(W—Ar), N(R)C(O)W—N(R4)2, or N(R)W—N(R4)2, wherein:each W is independently a valence bond or a C1-6alkylidene chain;R′ is an optionally substituted C1-6aliphatic group;each Ar1is independently selected from an optionally substituted 5-7 membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each R4is independently selected from R, COR5, CO2R5, CON(R5)2, SO2R5, SO2N(R5)2, or Ar1; andeach R5is independently selected from R or Ar.

Preferred R1groups of formula III include those described above for compounds of formula I.

Most preferably, the R3group of formula III is selected from CH2CH2NH2.

According to another embodiment, the present invention relates to a compound of formula IV:

or a pharmaceutically acceptable salt thereof, wherein A, B, R1, R3, W, and Ar are as defined above for compounds of formula I. Preferred A, B, R1, R3, W, and Ar groups of formula IV are those set forth above for compounds of formula I.

According to another embodiment, the present invention relates to a compound of formula V:

or a pharmaceutically acceptable salt thereof, wherein A, B, R1, R3, W, and Ar are as defined above for compounds of formula I. Preferred A, B, R1, R3, W, and Ar groups of formula V are those set forth above for compounds of formula I.

According to one embodiment, the present invention relates to a compound of formula I, wherein A and B are each —CH2—.

According to another embodiment, the present invention relates to a compound of formula II, wherein A and B are each —CH2—.

According to another embodiment, the present invention relates to a compound of formula III, wherein A and B are each —CH2—.

According to yet another embodiment, the present invention relates to a compound of formula IV, wherein A and B are each —CH2—.

According to another embodiment, the present invention relates to a compound of formula V, wherein A and B are each —CH2—.

According to one embodiment, the present invention relates to a compound of formula I, wherein A and B are each —CH2CH2—.

According to another embodiment, the present invention relates to a compound of formula II, wherein A and B are each —CH2CH2—.

According to another embodiment, the present invention relates to a compound of formula III, wherein A and B are each —CH2CH2—.

According to yet another embodiment, the present invention relates to a compound of formula IV, wherein A and B are each —CH2CH2—.

According to another embodiment, the present invention relates to a compound of formula V, wherein A and B are each —CH2CH2—.

According to one embodiment, the present invention relates to a compound of formula I, wherein one of A or B is —CH2— and the other of A or B is —CH2CH2—.

According to another embodiment, the present invention relates to a compound of formula II, wherein one of A or B is —CH2— and the other of A or B is —CH2CH2—.

According to another embodiment, the present invention relates to a compound of formula III, wherein one of A or B is —CH2— and the other of A or B is —CH2CH2—.

According to yet another embodiment, the present invention relates to a compound of formula IV, wherein one of A or B is —CH2— and the other of A or B is —CH2CH2—.

According to another embodiment, the present invention relates to a compound of formula V, wherein one of A or B is —CH2— and the other of A or B is —CH2CH2—.

Representative compounds of formula I are set forth in Table 1 below.

The compounds of the present invention may be prepared as illustrated by the Schemes I, II, and III below, by the Synthetic Examples described herein, and by general methods known to those of ordinary skill in the art.

Scheme I above shows a method for preparing tetrahydro-pyrrolo[3,4-c]pyrazoles. The tetrahydro-pyrrolo[3,4-c]pyrazoles 2 can be prepared in 5 steps from 4-acryloyl-benzoic acid methyl ester 1 by methods substantially similar to that described by Kikuchi, K. et. al.,J. Med. Chem.,2000, 43, 409-419.

Scheme II above shows an alternative method for preparing tetrahydro-pyrrolo[3,4-c]pyrazoles. The formation of the tetrahydro-pyrrolo[3,4-c]pyrazole 4 is achieved in 4 steps from 3. 3-Amino-5,6-dihydro-1H-pyrrolo[3,4-c]pyrazol-4-one 3 is synthesized in a manner substantially similar to that described by Gelin, S. et al.,Synth. Commun.,1982, 12 (6), 431-437.

Scheme III above shows a general method for preparing tetrahydro-pyrrolo[3,4-c]pyrazoles 5.

Accordingly, another embodiment of this invention provides a process for preparing a compound of this invention according to the methods of Schemes I, II, or III.

The activity of a compound utilized in this invention as an inhibitor of AKT or PDK1 kinase may be assayed in vitro, in vivo or in a cell line according to methods known in the art. In vitro assays include assays that determine inhibition of either the phosphorylation activity or ATPase activity of activated AKT or PDK1. Alternate in vitro assays quantitate the ability of the inhibitor to bind to AKT or PDK1. Inhibitor binding may be measured by radiolabelling the inhibitor prior to binding, isolating the inhibitor/AKT or inhibitor/PDK1 complex and determining the amount of radiolabel bound. Alternatively, inhibitor binding may be determined by running a competition experiment where compounds are incubated with AKT or PDK1 bound to known radioligands. Detailed conditions for assaying a compound utilized in this invention as an inhibitor of AKT or PDK1 kinase are set forth in the Examples below.

According to another embodiment, the invention provides a composition comprising a compound of this invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in the compositions of this invention is such that is effective to measurably inhibit a protein kinase, particularly AKT or PDK1 kinase, in a biological sample or in a patient. Preferably the composition of this invention is formulated for administration to a patient in need of such composition. Most preferably, the composition of this invention is formulated for oral administration to a patient.

The term “measurably inhibit”, as used herein means a measurable change in AKT or PDK1 activity between a sample comprising said composition and an AKT or PDK1 kinase and an equivalent sample comprising AKT or PDK1 kinase in the absence of said composition.

A “pharmaceutically acceptable salt” means any non-toxic salt or salt of an ester of a compound of this invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof. As used herein, the term “inhibitorily active metabolite or residue thereof” means that a metabolite or residue thereof is also an inhibitor of an AKT or PDK1 family kinase.

Salts derived from appropriate bases include alkali metal (e.g., sodium and potassium), alkaline earth metal (e.g., magnesium), ammonium and N+(C1-4alkyl)4salts. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization.

Most preferably, the pharmaceutically acceptable compositions of this invention are formulated for oral administration.

Depending upon the particular condition, or disease, to be treated or prevented, additional therapeutic agents, which are normally administered to treat or prevent that condition, may also be present in the compositions of this invention. As used herein, additional therapeutic agents that are normally administered to treat or prevent a particular disease, or condition, are known as “appropriate for the disease, or condition, being treated”.

For example, chemotherapeutic agents or other anti-proliferative agents may be combined with the compounds of this invention to treat proliferative diseases and cancer. Examples of known chemotherapeutic agents include, but are not limited to, Gleevec™, adriamycin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, taxol, interferons, and platinum derivatives.

Other examples of agents the inhibitors of this invention may also be combined with include, without limitation: treatments for Alzheimer's Disease such as Aricept® and Excelon®; treatments for Parkinson's Disease such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole, bromocriptine, pergolide, trihexephendyl, and amantadine; agents for treating Multiple Sclerosis (MS) such as beta interferon (e.g., Avonex® and Rebif®), Copaxone®, and mitoxantrone; treatments for asthma such as albuterol and Singulair®; agents for treating schizophrenia such as zyprexa, risperdal, seroquel, and haloperidol; anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA, azathioprine, cyclophosphamide, and sulfasalazine; immunomodulatory and immunosuppressive agents such as cyclosporin, tacrolimus, rapamycin, mycophenolate mofetil, interferons, corticosteroids, cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonian agents; agents for treating cardiovascular disease such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers, and statins; agents for treating liver disease such as corticosteroids, cholestyramine, interferons, and anti-viral agents; agents for treating blood disorders such as corticosteroids, anti-leukemic agents, and growth factors; and agents for treating immunodeficiency disorders such as gamma globulin.

According to another embodiment, the invention relates to a method of inhibiting AKT kinase activity in a biological sample comprising the step of contacting said biological sample with a compound of this invention, or a composition comprising said compound. Preferably, the method comprises the step of contacting said biological sample with a preferred compound of the present invention, as described herein supra.

According to another embodiment, the invention relates to a method of inhibiting PDK1 kinase activity in a biological sample comprising the step of contacting said biological sample with a compound of this invention, or a composition comprising said compound. Preferably, the method comprises the step of contacting said biological sample with a preferred compound of the present invention, as described herein supra.

Inhibition of AKT or PDK1 kinase activity in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ-transplantation, biological specimen storage, and biological assays.

Another aspect of this invention relates to a method for treating an AKT-mediated disease in a patient, which method comprises administering to a patient in need thereof, a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable composition comprising said compound. According to a preferred embodiment, the invention relates to administering a preferred compound of formula I, or a pharmaceutically acceptable composition comprising said compound.

Another aspect of this invention relates to a method for treating a PDK1-mediated disease in a patient, which method comprises administering to a patient in need thereof, a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable composition comprising said compound. According to a preferred embodiment, the invention relates to administering a preferred compound of formula I, or a pharmaceutically acceptable composition comprising said compound.

According to another embodiment, the present invention relates to a method for treating an AKT- or PDK1-mediated disease in a patient, which method comprises administering to a patient in need thereof, a therapeutically effective amount of a compound of formula II, III, IV, or V, or a pharmaceutically acceptable composition comprising said compound. According to another embodiment, said method comprises administering to a patient in need thereof, a therapeutically effective amount of a preferred compound of formula II, III, IV, or V, as described herein supra, or a pharmaceutically acceptable composition comprising said compound.

According to another embodiment, the present invention relates to a method for treating an AKT- or PDK1-mediated disease in a patient, which method comprises administering to a patient in need thereof, a therapeutically effective amount of a compound of formula IV or V, or a pharmaceutically acceptable composition comprising said compound. According to another embodiment, said method comprises administering to a patient in need thereof, a therapeutically effective amount of a preferred compound of formula IV, or V, as described herein supra, or a pharmaceutically acceptable composition comprising said compound.

According to another embodiment, the invention provides a method for treating or lessening the severity of an AKT-mediated disease or condition in a patient comprising the step of administering to said patient a composition according to the present invention.

The term “AKT-mediated condition” or “disease”, as used herein, means any disease or other deleterious condition in which AKT is known to play a role. The term “AKT-mediated condition” or “disease” also means those diseases or conditions that are alleviated by treatment with an AKT inhibitor. AKT-mediated diseases or conditions include, but are not limited to, proliferative disorders, cancer, cardiovascular disorders, rheumatoid arthritis, and neurodegenerative disorders. Preferably, said cancer is selected from pancreatic, prostate, or ovarian cancer.

According to another embodiment, the invention provides a method for treating or lessening the severity of an PDK1-mediated disease or condition in a patient comprising the step of administering to said patient a composition according to the present invention.

The term “PDK1-mediated condition” or “disease”, as used herein, means any disease or other deleterious condition in which PDK1 is known to play a role. The term “PDK1-mediated condition” or “disease” also means those diseases or conditions that are alleviated by treatment with a PDK1 inhibitor. PDK1-mediated diseases or conditions include, but are not limited to, proliferative disorders, and cancer. Preferably, said cancer is selected from brain (gliomas), breast, colon, head and neck, kidney, lung, liver, melanoma, ovarian, pancreatic, prostate, sarcoma, or thyroid.

According to another embodiment, the present invention relates to a method for treating or lessening the severity of a disease or condition selected from a proliferative disorder, a cardiac disorder, an inflammatory disorder, an autoimmune disorder, a viral disease, or a bone disorder, wherein said method comprises the step of administering an effective amount of a compound of the present invention. Preferably, said method comprises the step of administering an effective amount of a preferred compound of the present invention.

Preferably, the present invention relates to a method for treating or lessening the severity of a cancer.

More preferably, the present invention relates to a method for treating or lessening the severity of a cancer selected from brain (gliomas), breast, colon, head and neck, kidney, lung, liver, melanoma, ovarian, pancreatic, prostate, sarcoma, or thyroid.

Most preferably, the present invention relates to a method for treating or lessening the severity of pancreatic, prostate, or ovarian cancer.

In an alternate embodiment, the methods of this invention that utilize compositions that do not contain an additional therapeutic agent, comprise the additional step of separately administering to said patient an additional therapeutic agent. When these additional therapeutic agents are administered separately they may be administered to the patient prior to, sequentially with or following administration of the compositions of this invention.

The compounds of this invention or pharmaceutical compositions thereof may also be incorporated into compositions for coating an implantable medical device, such as prostheses, artificial valves, vascular grafts, stents and catheters. Vascular stents, for example, have been used to overcome restenosis (re-narrowing of the vessel wall after injury). However, patients using stents or other implantable devices risk clot formation or platelet activation. These unwanted effects may be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition comprising a compound of this invention. Suitable coatings and the general preparation of coated implantable devices are described in U.S. Pat. Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings are typically biocompatible polymeric materials such as a hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. The coatings may optionally be further covered by a suitable topcoat of fluorosilicone, polysaccarides, polyethylene glycol, phospholipids or combinations thereof to impart controlled release characteristics in the composition. Implantable devices coated with a compound of this invention are another embodiment of the present invention.

Compounds are screened for their ability to inhibit AKT using a standard coupled enzyme assay (Fox et al., Protein Sci., (1998) 7, 2249). Assays are carried out in a mixture of 100 mM HEPES 7.5, 10 mM MgCl2, 25 mM NaCl, 1 mM DTT and 3% DMSO. Final substrate concentrations in the assay are 170 μM ATP (Sigma Chemicals) and 200 μM peptide. Assays are carried out at 30° C. and 45 nM AKT. Final concentrations of the components of the coupled enzyme system are 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30 μg/ML pyruvate kinase and 10 μg/ml lactate dehydrogenase.

An assay stock buffer solution is prepared containing all of the reagents listed above, with the exception of AKT, DTT, and the test compound of interest. 55 μl of the stock solution is placed in a 96 well plate followed by addition of 2 μl of 1 mM DMSO stock containing the test compound (final compound concentration 30 μM). The plate is pre-incubated for about 10 minutes at 30° C. and the reaction initiated by addition of 10 μl of enzyme (final concentration 45 nM) and 1 mM DTT. Rates of reaction are obtained using a Molecular Devices SpectraMax Plus plate reader over a 15 minute read time at 30° C. Compounds showing greater than 50% inhibition versus standard wells containing the assay mixture and DMSO without test compound are titrated to determine IC50values.

Compounds are screened for their ability to inhibit PDK-1 using a radioactive-phosphate incorporation assay (Pitt and Lee, J. Biomol. Screen., (1996) 1, 47). Assays are carried out in a mixture of 100 mM HEPES (pH 7.5), 10 mM MgCl2, 25 mM NaCl, 2 mM DTT. Final substrate concentrations in the assay are 40 μM ATP (Sigma Chemicals) and 65 μM peptide (PDKtide, Upstate, Lake Placid, N.Y.). Assays are carried out at 30° C. and 25 nM PDK-1 in the presence of ˜27.5 nCi/μL of [γ-32P]ATP (Amersham Pharmacia Biotech, Amersham, UK). An assay stock buffer solution is prepared containing all of the reagents listed above, with the exception of ATP, and the test compound of interest. 15 μl of the stock solution is placed in a 96 well plate followed by addition of 1 μl of 0.5 mM DMSO stock containing the test compound (final compound concentration 25 μM, final DMSO concentration 5%). The plate is preincubated for about 10 minutes at 30° C. and the reaction initiated by addition of 4 μl ATP (final concentration 40 μM).

The reaction is stopped after 10 minutes by the addition of 100 μL 100 mM phosphoric acid, 0.01% Tween-20. A phosphocellulose 96 well plate (Millipore, Cat no. MAPHNOB50) is pretreated with 100 μL 100 mM phosphoric acid, 0.01% Tween-20 prior to the addition of the reaction mixture (100 μL). The spots are left to soak for at least 5 minutes, prior to wash steps (4×200 μL 100 mM phosphoric acid, 0.01% Tween-20). After drying, 20 μL Optiphase ‘SuperMix’ liquid scintillation cocktail (Perkin Elmer) is added to the well prior to scintillation counting (1450 Microbeta Liquid Scintillation Counter, Wallac).

Compounds showing greater than 50% inhibition versus standard wells containing the assay mixture and DMSO without test compound are titrated to determine IC50values.

The entire disclosure of all documents cited herein are incorporated herein by reference.