WO2013139882A1 - Pyrazolopyridines pour le traitement de la maladie de parkinson - Google Patents

Pyrazolopyridines pour le traitement de la maladie de parkinson Download PDF

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Publication number
WO2013139882A1
WO2013139882A1 PCT/EP2013/055878 EP2013055878W WO2013139882A1 WO 2013139882 A1 WO2013139882 A1 WO 2013139882A1 EP 2013055878 W EP2013055878 W EP 2013055878W WO 2013139882 A1 WO2013139882 A1 WO 2013139882A1
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pyridin
pyrazolo
amine
compound
formula
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PCT/EP2013/055878
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English (en)
Inventor
Bryan Chan
Anthony Estrada
Daniel Shore
Zachary Sweeney
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F. Hoffmann-La Roche Ag
Genentech, Inc.
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Publication of WO2013139882A1 publication Critical patent/WO2013139882A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs

Definitions

  • the present invention relates to pyrazolopyridine compounds that are capable of inhibiting one or more kinases, more particularly, LRRK2.
  • the compounds find applications in the treatment of a variety of disorders, including cancer and neurodegenerative diseases such as Parkinson's disease.
  • LRRK2 enzyme The defining feature of the LRRK2 enzyme is a Leucine Rich Repeat (LRR) motif (residues 1010-1291), a Ras-like small GTPase (residues 1336-1510), a region of high amino acid conservation that has been termed the C-terminal Of Ras of complex (COR) domain (residues 1511-1878), a protein kinase catalytic domain (residues 1879-2132) and a C-terminal WD40 motif (2231- 2276) [6, 7].
  • LRR Leucine Rich Repeat
  • the protein kinase domain of LRRK2 belongs to the tyrosine-like serine/threonine protein kinases and is most similar to the kinase RIP (Receptor Interacting Protein), which play key roles in innate immunity signalling pathways [8].
  • kinase RIP Receptor Interacting Protein
  • Moesin is a member of the Ezrin/Radixin/Moesin (ERM) family of proteins which functions to anchor the actin cytoskeleton to the plasma membrane and plays an important role in regulating membrane structure and organization [15, 16]. It was found that LRRK2 phosphorylated moesin at Thr558 [11], a previously characterised physiologically relevant phosphorylation site [15, 16]. LRRK2 also phosphorylated ezrin and radixin at the equivalent Thr residue.
  • EEM Ezrin/Radixin/Moesin
  • a first aspect of the invention relates to a compound of formula (IA) or (IB), or a pharmaceutically acceptable salt thereof,
  • R 1 is a group selected from alkyl, monocyclic heterocycloalkyl, bicyclic heterocycloalkyl and cycloalkyl, each of which is optionally substituted by one or more groups selected from alkyl, halo and cycloalkyl; where for formula (1 A): one, two or three of Xi, X 2 , X 3 and X4 are N, and the remainder are each independently CR 2 ; or
  • Xi, X 2 , X 3 and X 4 are each independently CR 2 ; or where for formula (IB):
  • X 4 is C orN; and one or two of Xi, X 2 and X 3 are independently selected from N and R , and the remainder are each independently CR 2 ; such that Xi, X 2 , X 3 , X4 and N form a heteroaryl group; each R 2 is independently selected from H, alkyl, CN, halo, heteroaryl, heterocycloalkyl, cycloalkyl, OR 4 , CONR 5 R 6 and S0 2 R 7 , wherein said alkyl, heteroaryl, heterocycloalkyl and cycloalkyl groups are each optionally further substituted by one or more groups selected from alkyl, halo and OR 9 ; each R is independently selected from H and alkyl, wherein said alkyl group is optionally further substituted by one or more groups selected from CN, halo, heteroaryl, heterocycloalkyl, cycloalkyl, OR 4 , CONR 5 R 6 and S0 2 R 7 ;
  • R 4 , R 5 , R 6 , R 7 and R 9 are each independently selected from H and alkyl; or R 5 and R 6 together with the nitrogen to which they are attached are linked to form a cyclic group which optionally further comprises one or more heteroatoms selected from O, N and S; with the proviso that when the compound is of formula (IB), the compound is other than 3-(5-isopropyl-4H-l,2,4-triazol-3-yl)-N-(tetrahydro-2H-pyran-4-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine or 3-(5-cyclopropyl-4H-l ,2,4-triazol-3-yl)-N-(tetrahydro-2H-pyran-4- yl)-lH-pyrazolo[4,3-c]pyridin-4-amine.
  • Another aspect of the invention relates to a compound of formula (1 A) or (IB), or a pharmaceutically acceptable salt thereof,
  • R 1 is a group selected from alkyl, monocyclic heterocycloalkyl, bicyclic heterocycloalkyl and cycloalkyl, each of which is optionally substituted by one or more groups selected from alkyl, halo and cycloalkyl; where for formula (1 A): one, two or three of Xi, X 2 , X 3 and X4 are N, and the remainder are each independently CR 2 ; or where for formula (IB): X 4 is C orN; and one or two of Xi, X 2 and X3 are independently selected from N and R 8 , and the remainder are each independently CR ; such that Xi, X 2 , ⁇ 3 ⁇ X4 and N form a heteroaryl group; each R 2 is independently selected from H, alkyl, CN, halo, heteroaryl, heterocycloalkyl, cycloalkyl, OR 4 , CONR 5 R 6 and S0 2 R 7 , wherein said alkyl
  • R 4 , R 5 , R 6 , R 7 and R 9 are each independently selected from H and alkyl; or
  • R 5 and R 6 together with the nitrogen to which they are attached are linked to form a cyclic group which optionally further comprises one or more heteroatoms selected from O, N and S; with the proviso that when the compound is of formula (IB), the compound is other than 3-(5-isopropyl-4H-l,2,4-triazol-3-yl)-N-(tetrahydro-2H-pyran-4-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine or 3-(5-cyclopropyl-4H-l ,2,4-triazol-3-yl)-N-(tetrahydro-2H-pyran-4- yl)-lH-pyrazolo[4,3-c]pyridin-4-amine.
  • a second aspect of the invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one compound as described above and a pharmaceutically acceptable carrier, diluent or excipient.
  • a third aspect of the invention relates to a compound as described above for use in medicine.
  • a fourth aspect of the invention relates to a compound as described above for use in treating a disorder selected from cancer and neurodegenerative diseases such as Parkinson's Disease.
  • a fifth aspect of the invention relates to the use of a compound as described above in the preparation of a medicament for treating or preventing a disorder selected from cancer and neurodegenerative diseases such as Parkinson's Disease.
  • a sixth aspect of the invention relates to the use of a compound as described above in the preparation of a medicament for the prevention or treatment of a disorder caused by, associated with or accompanied by any abnormal kinase activity wherein the kinase is preferably LRRK, more preferably LRRK2.
  • a seventh aspect of the invention relates to a method of treating a mammal having a disease state alleviated by inhibition of a kinase (preferably LRRK, more preferably LRRK2), wherein the method comprises administering to a mammal a therapeutically effective amount of a compound as described above.
  • An eighth aspect of the invention relates to the use of a compound as described above in an assay for identifying further candidate compounds capable of inhibition of a kinase, preferably LRRK, more preferably LRRK2.
  • a ninth aspect of the invention relates to a process for preparing a compound as described above.
  • the present invention relates to pyrazolopyridine compounds that are capable of inhibiting one or more kinases, more particularly LRRK, even more particularly LRRK2. Specifically, the invention relates to substituted pyrazolo[4,3-c]pyridine derivatives.
  • the presently claimed compounds are unsubstituted at the 7-position of the pyrazolo[4,3-c]pyridine core; (ii) have an amine group at the 4-position; and (iii) have an optionally substituted heteroaryl group at the 3-position oriented such that a hydrogen bond may form between a nitrogen of the heteroaryl group and the hydrogen of the amine group in the 4-position.
  • the hydrogen bonding arrangement is shown below:
  • the presently claimed compounds exhibit unexpectedly high LRRK2 affinity compared to compounds described in the art to date.
  • this particular arrangement of hydrogen bonding and substitution provides unexpectedly higher affinity values compared to analogous compounds that are substituted at the 7-position and/or have a heteroaryl in the 3-position that is not oriented in the correct position to form a hydrogen bond with the hydrogen of the amine group in the 4-position.
  • Alkyl is defined herein as a straight-chain or branched alkyl radical, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl or hexyl. Specific examples are methyl, ethyl and isopropyl.
  • the alkyl group is a Ci -20 alkyl group, more preferably a C M S, more preferably still a C1-12 alkyl group, more preferably still, a Ci-io alkyl group, more preferably a Ci-6 alkyl group or a C1-3 alkyl group.
  • a particular "alkyl” group is Ci- 6 -alkyl.
  • Cycloalkyl is defined herein as a monocyclic alkyl ring, such as, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl, or a fused bicyclic ring system such as norbomane. Specific examples are cyclopropyl and cyclohexyl.
  • the cycloalkyl group is a C3-12 cycloalkyl group, more preferably, a C3- 6 cycloalkyl group.
  • Halogen or halo is defined herein as chloro, fluoro, bromo or iodo.
  • a specific “halo” is chloro (CI).
  • aryl refers to a C 6 -i2 aromatic group, which may be benzocondensed, for example, phenyl or naphthyl.
  • Heteroaryl is defined herein as a monocyclic or bicyclic C 2-12 aromatic ring comprising one or more heteroatoms (that may be the same or different), such as oxygen, nitrogen or sulphur.
  • suitable heteroaryl groups include thienyl, furanyl, pyrrolyl, pyridinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl etc.
  • Heterocycloalkyl refers to a cyclic aliphatic group containing one or more heteroatoms selected from nitrogen, oxygen and sulphur, which is optionally interrupted by one or more -(CO)- groups in the ring and/or which optionally contains one or more double bonds in the ring.
  • the heterocycloalkyl group is a C3_7-heterocycloalkyl, more preferably a C3_6-heterocycloalkyl.
  • the heterocycloalkyl group is a C4_7-heterocycloalkyl, more preferably a C4_6-heterocycloalkyl.
  • Preferred heterocycloalkyl groups include, but are not limited to, piperazinyl, piperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl, tetrahydrofuranyl and tetrahydropyranyl. Specific examples are azetidinyl, morpholinyl, tetrahydropyranyl, tetrahydrofuranyl, pyrrolidinyl and piperidinyl.
  • the compound of the invention is of formula (IB), but with the proviso that when X and X are N, then X is other than carbon substituted with isopropyl or cyclopropyl.
  • the invention relates to a compound of formula (1 A), wherein X 4 is N and Xi, X 2 and X 3 are each independently CR 2 .
  • the invention relates to a compound of formula (1A), wherein X 2 is N and Xi, X 3 and X 4 are each independently CR 2 .
  • the invention relates to a compound of formula (1A), wherein Xi, X 2 , X 3 and X4 are each independently CR . In one preferred embodiment, the invention relates to a compound of formula (1A) wherein X 4 is N, Xi and X 2 are CH, and X 3 is CR 2 .
  • the invention relates to a compound of formula (1A) wherein X 2 is N, Xi and X 4 are CH, and X 3 is CR 2 . In one preferred embodiment, the invention relates to a compound of formula (1A) wherein Xi, X 2 and X4 are CH and X 3 is CR 2 .
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 and R 2 is H, methyl or morpholinyl. In one preferred embodiment the invention relates to a compound of formula (1A)
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 and R 2 is methyl.
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 and R 2 is morpholinyl.
  • the invention relates to a compound of formula (1A)
  • Xi is CR and R is H, methyl or morpholinyl and X 2 is CH.
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 and R 2 is H, methyl or morpholinyl and X 2 is N.
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 wherein R 2 is H, methyl or morpholinyl, X 2 is CH and X 3 is CR 2 wherein R 2 is (3R,4R)-3,4-difiuoropyrrolidin-l-yl, 1 ,3 -dimethyl- lH-pyrazol-4-yl, 1 ,5- dimethyl-lH-pyrazol-4-yl, 1 -methyl- lH-pyrazol-3-yl, 1 -methyl- lH-pyrazol-4-yl, 1- methyl-lH-pyrazol-5-yl, l-methylpiperidin-4-yl, 2,2,6,6-tetrafluoromorpholino, 2-fiuor
  • the invention relates to a compound of formula (1A)
  • Xi is CR wherein R is H, methyl or morpholinyl
  • X 2 is CH
  • X3 is CH
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 wherein R 2 is H, methyl or morpholinyl, X 2 is CH
  • X3 is CR 2 wherein R 2 is (3R,4R)-3,4-difluoropyrrolidin-l-yl, l ,3-dimethyl-lH-pyrazol-4-yl, 1 ,5 -dimethyl- 1H- pyrazol-4-yl, 1 -methyl- lH-pyrazol-3-yl, 1 -methyl- lH-pyrazol-4-yl, 1 -methyl- lH-pyrazol- 5-yl, l-methylpiperidin-4-yl, 2,2,6,6-tetrafluoromorpholino, 2-fluoro-pyridin-3-yl,
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 wherein R 2 is H, methyl or nK ⁇ holinyl, X 2 is CH, X 3 is CH and X 4 is N.
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 wherein R 2 is H, methyl or morpholinyl, X 2 is CH, X 3 is CR 2 wherein R 2 is (3R,4R)-3,4-difluoropyrrolidin-l-yl, l,3-dimethyl-lH-pyrazol-4-yl, 1 ,5 -dimethyl- 1H- pyrazol-4-yl, 1 -methyl- lH-pyrazol-3-yl, 1 -methyl- lH-pyrazol-4-yl, 1 -methyl- lH-pyrazol- 5-yl, l-methylpiperidin-4-yl, 2,2,6,6-tetrafluoromorpholino, 2-fiuoro-pyridin-3-yl, -C(O)- azetidin-l-yl, -C(0)-n ⁇ holinyl, -C
  • the invention relates to a compound of formula (1A) wherein Xi is CR 2 wherein R 2 is H, methyl or nK ⁇ holinyl, X 2 is CH, X 3 is CH and X 4 is CH.
  • the invention relates to a compound of formula (IB), wherein X 4 is C, Xi is NR 8 , and X 2 and X 3 are each independently CR .
  • the invention relates to a compound of formula (IB), wherein X 4 is C, X 3 is CR 2 , one of Xi and X 2 is N and the other is NR 8 .
  • the invention relates to a compound of formula (IB), wherein X 4 is C, X 2 is NR 8 , and Xi and X 3 are each independently CR .
  • the invention relates to a compound of formula (IB), wherein X 4 is N, and Xi, X 2 and X 3 are each independently CR 2 . In one preferred embodiment, the invention relates to a compound of formula (IB), wherein X 4 is C, X 2 is CR 2 , one of Xi and X 3 is N and the other is NR 8 .
  • the invention relates to a compound of formula (IB), wherein X 4 is C, Xi is NR 8 , and X 3 is CH andX 2 is CR 2 . In one preferred embodiment, the invention relates to a compound of formula (IB), wherein X 4 is C, X 3 is CH, Xi is N and X 2 is NR 8 .
  • the invention relates to a compound of formula (IB), wherein X 4 is C, X 2 is NR 8 , and Xi and X 3 are each independently CH.
  • the invention relates to a compound of formula (IB), wherein X 4 is N, Xi, and X 3 are CH and X 2 is CR 2 .
  • the invention relates to a compound of formula (IB), wherein X 4 is C, X 2 is CR 2 , one of Xi and X 3 is N and the other is NR 8 .
  • the invention relates to a compound of formula (IB) wherein Xi is CH, X 2 is CF 3 , X 3 is CH and X 4 is N. In one preferred embodiment the invention relates to a compound of formula (IB) wherein Xi is N, X 2 is C-cyclopropyl, X 3 is N-CH 3 and X 4 is C.
  • the invention relates to a compound of formula (IB) wherein Xi is N-CH 3 , X 2 is C-cyclopropyl or C-ethyl, X 3 is N or CH and X4 is C.
  • the invention relates to a compound of formula (IB) wherein Xi is N-CH 3 , X 2 is C-cyclopropyl, X 3 is N and X 4 is C.
  • the invention relates to a compound of formula (IB) wherein Xi is N-CH 3 , X 2 is C-cyclopropyl, X 3 is CH and X 4 is C.
  • the invention relates to a compound of formula (IB) wherein Xi is N-CH 3 , X 2 is C-ethyl, X 3 is N and X is C. In one preferred embodiment the invention relates to a compound of formula (IB) wherein Xi is N-CH 3 , X 2 is C-ethyl, X 3 is CH and X4 is C.
  • R 1 is:
  • R 1 is:
  • the invention relates to a compound of formula (1A) wherein R 1 is 2,2,2 -trifiuoroethyl, 2,6-dimethyltetrahydro-2H-pyran-4-yl, 4,4- difiuorocyclohexyl, 8-oxabicyclo[3.2.1]octan-3-yl, cyclopropylmethyl, dicyclopropylmethyl, ethyl, isopropyl or tetrahydro-2H-pyran-4-yl.
  • the invention relates to a compound of formula (1A) wherein R 1 is 2,2,2 -trifiuoroethyl.
  • the invention relates to a compound of formula (1A) wherein R 1 is 2,6-dimethyltetrahydro-2H-pyran-4-yl. In one preferred embodiment the invention relates to a compound of formula (1A) wherein R 1 is 4,4-difluorocyclohexyl.
  • the invention relates to a compound of formula (1A) wherein R 1 is 8-oxabicyclo[3.2.1]octan-3-yl. In one preferred embodiment the invention relates to a compound of formula (1A) wherein R 1 is cyclopropylmethyl.
  • the invention relates to a compound of formula (1A) wherein R 1 is dicyclopropylmethyl.
  • the invention relates to a compound of formula (1A) wherein R 1 is ethyl.
  • the invention relates to a compound of formula (1A) wherein R 1 isopropyl.
  • the invention relates to a compound of formula (1A) wherein R 1 is tetrahydro-2H-pyran-4-yl. In one preferred embodiment the invention relates to a compound of formula (IB) wherein R 1 is ethyl, isopropyl or tetrahydro-2H-pyran-4-yl.
  • the invention relates to a compound of formula (IB) wherein R 1 is ethyl.
  • the invention relates to a compound of formula (IB) wherein R 1 isopropyl.
  • the invention relates to a compound of formula (IB) wherein R 1 is tetrahydro-2H-pyran-4-yl.
  • R is:
  • R is: (i) a methyl, ethyl or isopropyl group (more preferably, a methyl or ethyl group) optionally substituted by one or more halo or OR 4 groups, where R 4 is alkyl;
  • a heteroaryl group selected from a pyrazolyl group and a pyridinyl group, each of which is optionally substituted by one or more alkyl or halo groups;
  • a heterocycloalkyl group selected from morpholinyl, tetrahydropyranyl, piperidinyl and tetrahydrofuranyl, each of which is optionally substituted by one or more alkyl or halo groups;
  • the compound of the invention is selected from the following:
  • the compound has a Ki value of less than about 0.1 ⁇ for LRRK2, more preferably, less than about 0.01 ⁇ , even more preferably, less than about 0.001 ⁇ .
  • the claimed compounds exhibit surprisingly good inhibition of LRRK2 (see Table 1), compared to compounds previously described in the art, for example, in WO 2010/106333 and WO 2011/141756.
  • LRRK2 see Table 1
  • Table 2 the structure and Ki values for various compounds described in WO 2010/106333 and WO 2011/141756 are shown in Table 2.
  • a further aspect of the invention relates to a compound as described above for use in medicine.
  • Another aspect of the invention relates to a compound as described above for use in treating cancer or a neurodegenerative disorder.
  • Another aspect relates to the use of a compound as described above in the preparation of a medicament for treating or preventing a neurodegenerative disorder.
  • the neurodegenerative disorder is Parkinson's Disease.
  • Another aspect relates to the use of a compound as described above in the preparation of a medicament for treating or preventing a proliferative disorder, for example, cancer.
  • the compound is administered in an amount sufficient to inhibit one or more kinases, preferably LRRK, even more preferably LRRK2.
  • Yet another aspect relates to the use of a compound of the invention in the preparation of a medicament for the prevention or treatment of a disorder caused by, associated with or accompanied by any abnormal activity against a biological target, wherein the target is a kinase, more preferably LRRK, even more preferably LRRK2.
  • the disorder is Parkinson's Disease.
  • Another aspect of the invention relates to a method of treating a protein kinase related disease or disorder.
  • the method according to this aspect of the present invention is effected by administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention, as described hereinabove, either per se, or, more preferably, as a part of a pharmaceutical composition, mixed with, for example, a pharmaceutically acceptable carrier, as is detailed hereinafter.
  • Yet another aspect of the invention relates to a method of treating a mammal having a disease state alleviated by inhibition of a protein kinase, wherein the method comprises administering to a mammal a therapeutically effective amount of a compound according to the invention.
  • the disease state is alleviated by the inhibition of the protein kinase LPvRK, more preferably LRRK2.
  • the mammal is a human.
  • method refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.
  • administering refers to a method for bringing a compound of the present invention and a protein kinase together in such a manner that the compound can affect the enzyme activity of the protein kinase either directly; i.e., by interacting with the protein kinase itself or indirectly; i.e., by interacting with another molecule on which the catalytic activity of the protein kinase is dependent.
  • administration can be accomplished either in vitro, i.e. in a test tube, or in vivo, i.e., in cells or tissues of a living organism.
  • the term “treating” includes abrogating, substantially inhibiting, slowing or reversing the progression of a disease or disorder, substantially ameliorating clinical symptoms of a disease or disorder or substantially preventing the appearance of clinical symptoms of a disease or disorder.
  • the term “preventing” refers to a method for barring an organism from acquiring a disorder or disease in the first place.
  • terapéuticaally effective amount refers to that amount of the compound being administered which will relieve to some extent one or more of the symptoms of the disease or disorder being treated.
  • a therapeutically effective amount can be estimated initially from cell culture assays.
  • a dose can be formulated in animal models to achieve a circulating concentration range that includes the IC50 or the IC100 as determined in cell culture. Such information can be used to more accurately determine useful doses in humans.
  • Initial dosages can also be estimated from in vivo data. Using these initial guidelines one of ordinary skill in the art could determine an effective dosage in humans.
  • toxicity and therapeutic efficacy of the compounds described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD 5 o and the ED 5 o.
  • the dose ratio between toxic and therapeutic effect is the therapeutic index and can be expressed as the ratio between LD 5 o and ED50.
  • Compounds which exhibit high therapeutic indices are preferred.
  • the data obtained from these cell cultures assays and animal studies can be used in formulating a dosage range that is not toxic for use in human.
  • the dosage of such compounds lies preferably within a range of circulating concentrations that include the ED 5 o with little or no toxicity.
  • the dosage may vary within this range depending upon the dosage form employed and the route of administration utilized.
  • the exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition, (see, e.g., Fingl et al, 1975, In: The Pharmacological Basis of Therapeutics, chapter 1, page 1).
  • Dosage amount and interval may be adjusted individually to provide plasma levels of the active compound which are sufficient to maintain therapeutic effect.
  • Usual patient dosages for oral administration range from about 50-2000 mg/kg/day, commonly from about 100-1000 mg/kg/day, preferably from about 150-700 mg/kg/day and most preferably from about 250-500 mg/kg/day.
  • therapeutically effective serum levels will be achieved by administering multiple doses each day.
  • the effective local concentration of the drug may not be related to plasma concentration.
  • One skilled in the art will be able to optimize therapeutically effective local dosages without undue experimentation.
  • kinase related disease or disorder refers to a disease or disorder characterized by inappropriate kinase activity or over-activity of a kinase as defined herein. Inappropriate activity refers to either; (i) kinase expression in cells which normally do not express said kinase; (ii) increased kinase expression leading to unwanted cell proliferation, differentiation and/or growth; or, (iii) decreased kinase expression leading to unwanted reductions in cell proliferation, differentiation and/or growth.
  • Over-activity of kinase refers to either amplification of the gene encoding a particular kinase or production of a level of kinase activity, which can correlate with a cell proliferation, differentiation and/or growth disorder (that is, as the level of the kinase increases, the severity of one or more of the symptoms of the cellular disorder increases).
  • Over activity can also be the result of ligand independent or constitutive activation as a result of mutations such as deletions of a fragment of a kinase responsible for ligand binding.
  • Preferred diseases or disorders that the compounds described herein may be useful in preventing include cancer and neurodegenerative disorders such as Parkinson's Disease.
  • the present invention further provides use of compounds as defined herein for the manufacture of medicaments for the treatment of diseases where it is desirable to inhibit LRRK2.
  • diseases include Parkinson's Disease.
  • the compounds or physiologically acceptable salt, ester or other physiologically functional derivative thereof, described herein may be presented as a pharmaceutical formulation, comprising the compounds or physiologically acceptable salt, ester or other physiologically functional derivative thereof, together with one or more pharmaceutically acceptable carriers therefore and optionally other therapeutic and/or prophylactic ingredients.
  • the carrier(s) must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
  • the pharmaceutical compositions may be for human or animal usage in human and veterinary medicine. Examples of such suitable excipients for the various different forms of pharmaceutical compositions described herein may be found in the "Handbook of Pharmaceutical Excipients, 2 nd Edition, (1994), Edited by A Wade and PJ Weller.
  • Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985).
  • suitable carriers include lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol and the like.
  • suitable diluents include ethanol, glycerol and water. The choice of pharmaceutical carrier, excipient or diluent can be selected with regard to the intended route of administration and standard pharmaceutical practice.
  • compositions may comprise as, or in addition to, the carrier, excipient or diluent any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s), solubilising agent(s), buffer(s), flavouring agent(s), surface active agent(s), thickener(s), preservative(s) (including anti-oxidants) and the like, and substances included for the purpose of rendering the formulation isotonic with the blood of the intended recipient.
  • any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s), solubilising agent(s), buffer(s), flavouring agent(s), surface active agent(s), thickener(s), preservative(s) (including anti-oxidants) and the like and substances included for the purpose of rendering the formulation isotonic with the blood of the intended recipient.
  • Suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, free-flow lactose, beta-lactose, corn sweeteners, natural and synthetic gums, such as acacia, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol.
  • Suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Preservatives, stabilizers, dyes and even flavoring agents may be provided in the pharmaceutical composition.
  • preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid.
  • Antioxidants and suspending agents may be also used.
  • compositions include those suitable for oral, topical (including dermal, buccal and sublingual), rectal or parenteral (including subcutaneous, intradermal, intramuscular and intravenous), nasal and pulmonary administration e.g., by inhalation.
  • the formulation may, where appropriate, be conveniently presented in discrete dosage units and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association an active compound with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.
  • compositions suitable for oral administration wherein the carrier is a solid are most preferably presented as unit dose formulations such as boluses, capsules or tablets each containing a predetermined amount of active compound.
  • a tablet may be made by compression or moulding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared by compressing in a suitable machine an active compound in a free-flowing form such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, lubricating agent, surface-active agent or dispersing agent.
  • Moulded tablets may be made by moulding an active compound with an inert liquid diluent. Tablets may be optionally coated and, if uncoated, may optionally be scored.
  • Capsules may be prepared by filling an active compound, either alone or in admixture with one or more accessory ingredients, into the capsule shells and then sealing them in the usual manner.
  • Cachets are analogous to capsules wherein an active compound together with any accessory ingredient(s) is sealed in a rice paper envelope.
  • An active compound may also be formulated as dispersible granules, which may for example be suspended in water before administration, or sprinkled on food. The granules may be packaged, e.g., in a sachet.
  • Formulations suitable for oral administration wherein the carrier is a liquid may be presented as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil- in -water liquid emulsion.
  • Formulations for oral administration include controlled release dosage forms, e.g., tablets wherein an active compound is formulated in an appropriate release - controlling matrix, or is coated with a suitable release - controlling film. Such formulations may be particularly convenient for prophylactic use.
  • compositions suitable for rectal administration wherein the carrier is a solid are most preferably presented as unit dose suppositories.
  • Suitable carriers include cocoa butter and other materials commonly used in the art.
  • the suppositories may be conveniently formed by admixture of an active compound with the softened or melted carrier(s) followed by chilling and shaping in moulds.
  • Pharmaceutical formulations suitable for parenteral administration include sterile solutions or suspensions of an active compound in aqueous or oleaginous vehicles.
  • Injectable preparations may be adapted for bolus injection or continuous infusion. Such preparations are conveniently presented in unit dose or multi-dose containers which are sealed after introduction of the formulation until required for use.
  • an active compound may be in powder form which is constituted with a suitable vehicle, such as sterile, pyrogen-free water, before use.
  • An active compound may also be formulated as long-acting depot preparations, which may be administered by intramuscular injection or by implantation, e.g., subcutaneously or intramuscularly.
  • Depot preparations may include, for example, suitable polymeric or hydrophobic materials, or ion-exchange resins. Such long-acting formulations are particularly convenient for prophylactic use.
  • Formulations suitable for pulmonary administration via the buccal cavity are presented such that particles containing an active compound and desirably having a diameter in the range of 0.5 to 7 microns are delivered in the bronchial tree of the recipient.
  • such formulations are in the form of finely comminuted powders which may conveniently be presented either in a pierceable capsule, suitably of, for example, gelatin, for use in an inhalation device, or alternatively as a self-propelling formulation comprising an active compound, a suitable liquid or gaseous propellant and optionally other ingredients such as a surfactant and/or a solid diluent.
  • suitable liquid propellants include propane and the chlorofiuorocarbons
  • suitable gaseous propellants include carbon dioxide.
  • Self-propelling formulations may also be employed wherein an active compound is dispensed in the form of droplets of solution or suspension.
  • Such self-propelling formulations are analogous to those known in the art and may be prepared by established procedures. Suitably they are presented in a container provided with either a manually-operable or automatically functioning valve having the desired spray characteristics; advantageously the valve is of a metered type delivering a fixed volume, for example, 25 to 100 microlitres, upon each operation thereof.
  • an active compound may be in the form of a solution or suspension for use in an atomizer or nebuliser whereby an accelerated airstream or ultrasonic agitation is employed to produce a fine droplet mist for inhalation.
  • Formulations suitable for nasal administration include preparations generally similar to those described above for pulmonary administration. When dispensed such formulations should desirably have a particle diameter in the range 10 to 200 microns to enable retention in the nasal cavity; this may be achieved by, as appropriate, use of a powder of a suitable particle size or choice of an appropriate valve. Other suitable formulations include coarse powders having a particle diameter in the range 20 to 500 microns, for administration by rapid inhalation through the nasal passage from a container held close up to the nose, and nasal drops comprising 0.2 to 5% w/v of an active compound in aqueous or oily solution or suspension.
  • Pharmaceutically acceptable carriers are well known to those skilled in the art and include, but are not limited to, 0.1 M and preferably 0.05 M phosphate buffer or 0.8% saline. Additionally, such pharmaceutically acceptable carriers may be aqueous or nonaqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.
  • Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media.
  • Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils. Preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, chelating agents, inert gases and the like.
  • Formulations suitable for topical formulation may be provided for example as gels, creams or ointments. Such preparations may be applied e.g. to a wound or ulcer either directly spread upon the surface of the wound or ulcer or carried on a suitable support such as a bandage, gauze, mesh or the like which may be applied to and over the area to be treated.
  • a suitable support such as a bandage, gauze, mesh or the like which may be applied to and over the area to be treated.
  • Liquid or powder formulations may also be provided which can be sprayed or sprinkled directly onto the site to be treated, e.g. a wound or ulcer.
  • a carrier such as a bandage, gauze, mesh or the like can be sprayed or sprinkle with the formulation and then applied to the site to be treated.
  • a process for the preparation of a pharmaceutical or veterinary composition as described above comprising bringing the active compound(s) into association with the carrier, for example by admixture.
  • the formulations are prepared by uniformly and intimately bringing into association the active agent with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product.
  • the invention extends to methods for preparing a pharmaceutical composition comprising bringing a compound of general formula (I) in conjunction or association with a pharmaceutically or veterinarily acceptable carrier or vehicle.
  • the compounds of the invention can be present as salts or esters, in particular pharmaceutically and veterinarily acceptable salts or esters.
  • salts of the compounds of the invention include suitable acid addition or base salts thereof.
  • suitable pharmaceutical salts may be found in Berge et al, J Pharm Sci, 66, 1-19 (1977). Salts are formed, for example with strong inorganic acids such as mineral acids, e.g.
  • hydrohalic acids such as hydrochloride, hydrobromide and hydroiodide, sulphuric acid, phosphoric acid sulphate, bisulphate, hemisulphate, thiocyanate, persulphate and sulphonic acids; with strong organic carboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atoms which are unsubstituted or substituted (e.g., by halogen), such as acetic acid; with saturated or unsaturated dicarboxylic acids, for example oxalic, malonic, succinic, maleic, fumaric, phthalic or tetraphthalic; with hydroxycarboxylic acids, for example ascorbic, glycolic, lactic, malic, tartaric or citric acid; with aminoacids, for example aspartic or glutamic acid; with benzoic acid; or with organic sulfonic acids, such as (Ci-C4)-alkyl- or aryl-sulfonic acids
  • Salts which are not pharmaceutically or veterinarily acceptable may still be valuable as intermediates.
  • Preferred salts include, for example, acetate, trifluoroacetate, lactate, gluconate, citrate, tartrate, maleate, malate, pantothenate, adipate, alginate, aspartate, benzoate, butyrate, digluconate, cyclopentanate, glucoheptanate, glycerophosphate, oxalate, heptanoate, hexanoate, fumarate, nicotinate, palmoate, pectinate, 3-phenylpropionate, picrate, pivalate, proprionate, tartrate, lactobionate, pivolate, camphorate, undecanoate and succinate, organic sulphonic acids such as methanesulphonate, ethanesulphonate, 2- hydroxyethane sulphonate, camphorsulphonate, 2-naphthalenesulphon
  • Esters are formed either using organic acids or alcohols/hydroxides, depending on the functional group being esterified.
  • Organic acids include carboxylic acids, such as alkanecarboxylic acids of 1 to 12 carbon atoms which are unsubstituted or substituted (e.g., by halogen), such as acetic acid; with saturated or unsaturated dicarboxylic acid, for example oxalic, malonic, succinic, maleic, fumaric, phthalic or tetraphthalic; with hydroxycarboxylic acids, for example ascorbic, glycolic, lactic, malic, tartaric or citric acid; with aminoacids, for example aspartic or glutamic acid; with benzoic acid; or with organic sulfonic acids, such as (Ci-C4)-alkyl- or aryl-sulfonic acids which are unsubstituted or substituted (for example, by a halogen) such as methane- or p-toluen
  • Suitable hydroxides include inorganic hydroxides, such as sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminium hydroxide.
  • Alcohols include alkanealcohols of 1-12 carbon atoms which may be unsubstituted or substituted, e.g. by a halogen).
  • the invention includes, where appropriate all enantiomers, diastereoisomers and tautomers of the compounds of the invention.
  • the person skilled in the art will recognise compounds that possess optical properties (one or more chiral carbon atoms) or tautomeric characteristics.
  • the corresponding enantiomers and/or tautomers may be isolated/prepared by methods known in the art.
  • Enantiomers are characterised by the absolute configuration of their chiral centres and described by the R- and ⁇ -sequencing rules of Cahn, Ingold and Prelog. Such conventions are well known in the art (e.g. see 'Advanced Organic Chemistry', 3 rd edition, ed.
  • Compounds of the invention containing a chiral centre may be used as a racemic mixture, an enantiomerically enriched mixture, or the racemic mixture may be separated using well-known techniques and an individual enantiomer may be used alone.
  • Some of the compounds of the invention may exist as stereoisomers and/or geometric isomers - e.g. they may possess one or more asymmetric and/or geometric centres and so may exist in two or more stereoisomeric and/or geometric forms.
  • the present invention contemplates the use of all the individual stereoisomers and geometric isomers of those inhibitor agents, and mixtures thereof.
  • the terms used in the claims encompass these forms, provided said forms retain the appropriate functional activity (though not necessarily to the same degree).
  • the present invention also includes all suitable isotopic variations of the agent or a pharmaceutically acceptable salt thereof.
  • An isotopic variation of an agent of the present invention or a pharmaceutically acceptable salt thereof is defined as one in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually found in nature.
  • isotopes that can be incorporated into the agent and pharmaceutically acceptable salts thereof include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine and chlorine such as 2 H, 3 H, 13 C, 14 C, 15 N, 17 0, 18 0, 31 P, 32 P, 35 S, 18 F and 36 C1, respectively.
  • isotopic variations of the agent and pharmaceutically acceptable salts thereof are useful in drug and/or substrate tissue distribution studies. Tritiated, i.e., 3 H, and carbon-14, i.e., 14 C, isotopes are particularly preferred for their ease of preparation and detectabihty. Further, substitution with isotopes such as deuterium, i.e., 2 H, 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.
  • the invention includes compounds of general formula (I) where any hydrogen atom has been replaced by a deuterium atom. Isotopic variations of the agent of the present invention and pharmaceutically acceptable salts thereof of this invention can generally be prepared by conventional procedures using appropriate isotopic variations of suitable reagents.
  • the invention further includes the compounds of the present invention in prodrug form, i.e. covalently bonded compounds which release the active parent drug according to general formula (I) in vivo.
  • prodrugs are generally compounds of the invention wherein one or more appropriate groups have been modified such that the modification may be reversed upon administration to a human or mammalian subject. Reversion is usually performed by an enzyme naturally present in such subject, though it is possible for a second agent to be administered together with such a prodrug in order to perform the reversion in vivo. Examples of such modifications include ester (for example, any of those described above), wherein the reversion may be carried out be an esterase etc. Other such systems will be well known to those skilled in the art.
  • the present invention also includes solvate forms of the compounds of the present invention.
  • the terms used in the claims encompass these forms.
  • POLYMORPHS The invention further relates to the compounds of the present invention in their various crystalline forms, polymorphic forms and (an)hydrous forms. It is well established within the pharmaceutical industry that chemical compounds may be isolated in any of such forms by slightly varying the method of purification and or isolation form the solvents used in the synthetic preparation of such compounds.
  • compositions of the present invention may be adapted for rectal, nasal, intrabronchial, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intraarterial and intradermal), intraperitoneal or intrathecal administration.
  • the formulation is an orally administered formulation.
  • the formulations may conveniently be presented in unit dosage form, i.e., in the form of discrete portions containing a unit dose, or a multiple or sub-unit of a unit dose.
  • the formulations may be in the form of tablets and sustained release capsules, and may be prepared by any method well known in the art of pharmacy.
  • Formulations for oral administration in the present invention may be presented as: discrete units such as capsules, gellules, drops, cachets, pills or tablets each containing a predetermined amount of the active agent; as a powder or granules; as a solution, emulsion or a suspension of the active agent in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water- in-oil liquid emulsion; or as a bolus etc.
  • these compositions contain from 1 to 250 mg and more preferably from 10-100 mg, of active ingredient per dose.
  • the term "acceptable carrier” includes vehicles such as common excipients e.g. binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (Povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose, hydroxypropyl- methylcellulose, sucrose and starch; fillers and carriers, for example com starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride and alginic acid; and lubricants such as magnesium stearate, sodium stearate and other metallic stearates, glycerol stearate stearic acid, silicone fluid, talc waxes, oils and colloidal silica.
  • Flavouring agents such as peppermint, oil of wintergreen, cherry flavouring and the like can also be used. It may
  • a tablet may be made by compression or moulding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared by compressing in a suitable machine the active agent in a free flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface-active or dispersing agent.
  • Moulded tablets may be made by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
  • the tablets may be optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active agent.
  • compositions suitable for oral administration include lozenges comprising the active agent in a flavoured base, usually sucrose and acacia or tragacanth; pastilles comprising the active agent in an inert base such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active agent in a suitable liquid carrier.
  • compositions or emulsions which may be injected intravenously, intraarterially, intrathecally, subcutaneously, intradermally, intraperitoneally or intramuscularly, and which are prepared from sterile or sterilisable solutions.
  • injectable forms typically contain between 10 - 1000 mg, preferably between 10 - 250 mg, of active ingredient per dose.
  • compositions of the present invention may also be in form of suppositories, pessaries, suspensions, emulsions, lotions, ointments, creams, gels, sprays, solutions or dusting powders.
  • An alternative means of transdermal administration is by use of a skin patch.
  • the active ingredient can be incorporated into a cream consisting of an aqueous emulsion of polyethylene glycols or liquid paraffin.
  • the active ingredient can also be incorporated, at a concentration of between 1 and 10% by weight, into an ointment consisting of a white wax or white soft paraffin base together with such stabilisers and preservatives as may be required.
  • a person of ordinary skill in the art can easily determine an appropriate dose of one of the instant compositions to administer to a subject without undue experimentation.
  • a physician will determine the actual dosage which will be most suitable for an individual patient and it will depend on a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the individual undergoing therapy.
  • the dosages disclosed herein are exemplary of the average case. There can of course be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention.
  • an effective amount of a compound of general formula (I) may be administered to inhibit the kinase implicated with a particular condition or disease.
  • this dosage amount will further be modified according to the type of administration of the compound.
  • parenteral administration of a compound of general formula (I) is preferred.
  • An intravenous infusion of the compound in 5% dextrose in water or normal saline, or a similar formulation with suitable excipients, is most effective, although an intramuscular bolus injection is also useful.
  • the parenteral dose will be about 0.01 to about 100 mg/kg; preferably between 0.1 and 20 mg/kg, in a manner to maintain the concentration of drug in the plasma at a concentration effective to inhibit a kinase.
  • the compounds may be administered one to four times daily at a level to achieve a total daily dose of about 0.4 to about 400 mg/kg/day.
  • the precise amount of an inventive compound which is therapeutically effective, and the route by which such compound is best administered, is readily determined by one of ordinary skill in the art by comparing the blood level of the agent to the concentration required to have a therapeutic effect.
  • the compounds of this invention may also be administered orally to the patient, in a manner such that the concentration of drug is sufficient to achieve one or more of the therapeutic indications disclosed herein.
  • a pharmaceutical composition containing the compound is administered at an oral dose of between about 0.1 to about 50 mg/kg in a manner consistent with the condition of the patient.
  • the oral dose would be about 0.5 to about 20 mg/kg.
  • the compounds of this invention which may have good bioavailability, may be tested in one of several biological assays to determine the concentration of a compound which is required to have a given pharmacological effect.
  • the one or more compounds of the invention are administered in combination with one or more other active agents, for example, existing drugs available on the market.
  • the compounds of the invention may be administered consecutively, simultaneously or sequentially with the one or more other active agents.
  • Drugs in general are more effective when used in combination.
  • combination therapy is desirable in order to avoid an overlap of major toxicities, mechanism of action and resistance mechanism(s).
  • the major advantages of combining chemotherapeutic drugs are that it may promote additive or possible synergistic effects through biochemical interactions and also may decrease the emergence of resistance.
  • Beneficial combinations may be suggested by studying the inhibitory activity of the test compounds with agents known or suspected of being valuable in the treatment of a particular disorder. This procedure can also be used to determine the order of administration of the agents, i.e. before, simultaneously, or after delivery. Such scheduling may be a feature of all the active agents identified herein.
  • a further aspect of the invention relates to the use of a compound as described above in an assay for identifying further candidate compounds capable of inhibiting one or more kinases, more preferably LRRK, even more preferably, LRRK2.
  • the assay is a competitive binding assay.
  • the competitive binding assay comprises contacting a compound of the invention with a kinase, preferably LRRK, more preferably LRRK2, and a candidate compound and detecting any change in the interaction between the compound according to the invention and the kinase.
  • a kinase preferably LRRK, more preferably LRRK2
  • the candidate compound is generated by conventional SAR modification of a compound of the invention.
  • conventional SAR modification refers to standard methods known in the art for varying a given compound by way of chemical derivatisation.
  • the identified compound may act as a model (for example, a template) for the development of other compounds.
  • the compounds employed in such a test may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. The abolition of activity or the formation of binding complexes between the compound and the agent being tested may be measured.
  • the assay of the present invention may be a screen, whereby a number of agents are tested.
  • the assay method of the present invention is a high through-put screen.
  • This invention also contemplates the use of competitive drug screening assays in which neutralising antibodies capable of binding a compound specifically compete with a test compound for binding to a compound.
  • HTS high throughput screening
  • the competitive binding assay comprises contacting a compound of the invention with a kinase in the presence of a known substrate of said kinase and detecting any change in the interaction between said kinase and said known substrate.
  • a further aspect of the invention provides a method of detecting the binding of a ligand to a kinase, said method comprising the steps of: a ligand with a kinase in the presence of a known detecting any change in the interaction between said kinase and said and wherein said ligand is a compound of the invention.
  • One aspect of the invention relates to a process comprising the steps of:
  • Another aspect of the invention provides a process comprising the steps of:
  • Another aspect of the invention provides a process comprising the steps of:
  • the invention also relates to a ligand identified by the method described hereinabove.
  • Yet another aspect of the invention relates to a pharmaceutical composition comprising a ligand identified by the method described hereinabove.
  • Another aspect of the invention relates to the use of a ligand identified by the method described hereinabove in the preparation of a pharmaceutical composition for use in the treatment of one or more disorders [insert list of disorders].
  • the above methods may be used to screen for a ligand useful as an inhibitor of one or more kinases.
  • Compounds of general formula (I) are useful both as laboratory tools and as therapeutic agents. In the laboratory certain compounds of the invention are useful in establishing whether a known or newly discovered kinase contributes a critical or at least significant biochemical function during the establishment or progression of a disease state, a process commonly referred to as 'target validation'.
  • a further aspect of the invention relates to a process for preparing a compound of formula (1A) or (IB) as defined above, said process comprising the steps of:
  • Suitable protecting groups, PG will be familiar to a person skilled in the art (see, for example, Greene, Theodora W. and Wuts, Peter G. M. Greene's Protective Groups in Organic Synthesis. 4th Ed. (2006)), and include, but are not limited to, tert- butoxycarbonyl, benzyloxycarbonyl, benzyl, 4-methoxybenzyl, 2,4-dimethoxybenzyl or trityl.
  • the protecting group, PG is trityl or 4- methoxybenzyl (PMB).
  • step (iii) comprises reacting said compound of formula (2) with 1,1,1,2,2,2-hexamethyldistannane and trans-Pd(PPri3)2Cl2 to form an intermediate of formula (2 A):
  • the coupling takes place in the presence of a palladium catalyst, more preferably, Pd(PPh3)4/CuI.
  • the compound of formula (2 A) is coupled with a compound of formula (5 A) or (5B),
  • the compound resulting from the above coupling of (2A) with (5 A) or (5B) is further modified to form a compound of formula (1 A) or (IB).
  • the invention is further described by way of the following non-limiting examples, and with reference to the following figures, wherein:
  • Figure 1 shows the domain structure of LRRK1 and local mutations that have been linked to Parkinson's disease.
  • All LRRK2 protein kinases were of human origin and were sourced from Invitrogen Corporation (Carlsbad, CA 92008 USA) unless otherwise indicated.
  • the active mutant used was recombinant human, catalytic domain (amino acids 970-2527) containing a G2019S mutation, GST-tagged, expressed in insect cells (Invitrogen Cat#PV4881).
  • the wild type used was recombinant human, catalytic domain (amino acids 970-2527) GST - tagged, expressed in insect cells (Invitrogen Cat#PV4873).
  • the kinase dead mutant used was recombinant human, catalytic domain (amino acids 970-2527) containing a D1994A mutation, GST-tagged, expressed in insect cells (Invitrogen Cat#PM4041AE). No special measures were taken to activate any of the kinases.
  • the assays were initiated with Mg/ATP and stopped by the addition of 25 ⁇ 1 ⁇ 11 50% orthophosphoric acid. Reactions were harvested onto Whatman P81 Unifilter Plates (Fisher Scientific. Loughborough, LE115RG, UK. Cat# FDU-105-020U) using a Tomtec harvester. (Tomtec Hamden, Ct 06514. USA) Plates were counted using a Perkin Elmer Top Count NX7. (Perkin Elmer, Shelton CT 06484-4794 USA)
  • IC50 values of inhibitors were determined after carrying out assays at 10 different concentrations of each compound in duplicate.
  • Preparative high pressure liquid chromatography was carried out using apparatus made by Agilent.
  • the apparatus is constructed such that the chromatography is monitored by a multi-wavelength UV detector (G1365B manufactured by Agilent) and an MM- ES+APCI mass spectrometer (G- 1956 A, manufactured by Agilent) connected in series, and if the appropriate criteria are met the sample is collected by an automated fraction collector (G1364B manufactured by Agilent). Collection can be triggered by any combination of UV or mass spectrometry or can be based on time.
  • Typical conditions for the separation process are as follows: The gradient is run over a 10 minute period (gradient at start: 10% methanol and 90% water, gradient at finish: 100% methanol and 0% water; as buffer: either 0.1%> trifluoroacetic acid is added to the water (low pH buffer), or ammonium bicarbonate (10 mmol / 1) and 35% ammonium hydroxide (1.6 ml / 1) is added to the water (high pH buffer).
  • buffer either 0.1%> trifluoroacetic acid is added to the water (low pH buffer), or ammonium bicarbonate (10 mmol / 1) and 35% ammonium hydroxide (1.6 ml / 1) is added to the water (high pH buffer).
  • Flash chromatography refers to silica gel chromatography and carried out using an SP4 or an Isolara 4 MPLC system (manufactured by Biotage); pre-packed silica gel cartridges (supplied by Biotage); or using conventional glass column chromatography.
  • Xantphos 4 ,5 -B is (diphenylpho sphino)-9 ,9 -dimethy l anthene
  • HATU N,N,N',N'-Tetramethyl-0-(7-azabenzotriazol- 1 -yl)uronium- hexafluorophospate
  • DIPEA N,N-diisopropylethylamine
  • TBSMSC1 Tertiarybutyldimethylsilyl chloride
  • TPAP Tetrapropylammonium perruthenate
  • Step 1 Synthesis of 4-chloro-3-iodo-l-trityl-lH-pyrazolo[4,3-c]pyridine: A suspension of 4-chloro-3-iodo-lH-pyrozolo[4,3-c]pyridine (7.88 g, 0.0280 mol) in DCM (100 mL) was stirred at 0 °C for 5 minutes. TEA (5.62 g, 0.0560 mol) was added and the mixture was stirred for 10 minutes. Trityl chloride (11.7 g, 0.0420 mol) was then added. After being stirred for 3 hours, the reaction mixture was treated with water (100 mL). The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound as an off-white solid (13.2 g, 89%).
  • Step 2 Synthesis of 3-iodo-N-isopropyl-l-trityl-lH-pyrazolo[4,3-c]pyridin-4- amine:
  • a microwave tube equipped with a magnetic stirrer was charged with 4-chloro-3- iodo-l-trityl-lH-pyrazolo[4,3-c]pyridine (2.00 g, 3.84 mmol), propan-2-amine (1.10 g, 19.2 mmol), n-BuOH (0.86 g, 7.68 mmol), and DIPEA (10 mL).
  • the reaction mixture was heated at 170 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • the residue was purified by silica gel column chromatography eluting with DCM/petroleum ether/TEA (2/1/0.01) to afford the title compound as a white solid (1.4 g, 67 %).
  • Step 3 Synthesis of N-isopropyl-3-(trimethylstannyl)-l-trityl-lH-pyrazolo[4,3- c]pyridin-4-amine: A reaction tube equipped with a magnetic stirrer was charged with 3- iodo-N-isopropyl-l-trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (1.00 g, 1.84 mmol), 1,1,1,2,2,2-hexamethyldistannane (723 mg, 2.21 mmol), toluene (12 mL), and trans- Pd(PPh 3 ) 2 Cl 2 (40 mg, 0.055 mmol).
  • Step 4 Synthesis of N-isopropyl-3-(pyridin-2-yl)-l-trityl-lH-pyrazolo[4,3- c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-isopropyl-3-(trimethylstannyl)-l-trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (100 mg, 0.172 mmol), 2-bromopyridine (54.0 mg, 0.344 mmol), LiCl (29.0 mg, 0.688 mmol), Cul (7.0 mg, 0.034 mmol), Pd(PPh 3 ) 4 (20.0 mg, 0.0172 mmol) and THF (2 mL).
  • reaction mixture was heated at 100 °C for 1 hour under microwave irradiation. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (1/2) to afford the title compound as a yellow solid (65 mg, 76%).
  • Step 5 Synthesis of N-isopropyl-3-(pyridin-2-yl)-lH-pyrazolo[4,3-c]pyridin-4- amine: To a mixture of N-isopropyl-3-(pyridin-2-yl)-l-trityl-lH-pyrazolo[4,3-c]pyridin-4- amine (65.0 mg, 0.131 mmol) in dichloromethane (5 mL) was added tnfluoroacetic acid (1.0 mL) and triethylsilane (0.20 mL). The mixture was stirred under reflux for 1 hour. After cooling down, the reaction mixture was neutralized to pH around 7 with saturated NaHC0 3 solution.
  • Step 1 Synthesis of 3-(4-chloropyridin-2-yl)-N-isopropyl-l-trityl-lH-pyrazolo[4,3- c]pyridin-4-amine: To a microwave tube was added N-isopropyl-3-(trimethylstannyl)-l- trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (150 mg, 0.258 mmol), 2-bromo-4- chloropyridine (99.0 mg, 0.515 mmol), LiCl (43.0 mg, 1.03 mmol), Cul (10.0 mg, 0.0515 mmol), Pd(PPh 3 ) 4 (30.0 mg, 0.0258 mmol) and THF (3.0 mL).
  • reaction mixture was heated at 100 °C for 1 hour under microwave irradiation. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure. The residue was purified by prep- TLC eluting with petroleum ether/ethyl acetate (1 :2) to afford the title compound as a yellow solid (100 mg, 73%).
  • Step 2 Synthesis of N-isopropyl-3-(4-morpholinopyridin-2-yl)-l-trityl-lH- pyrazolo[4,3-c]pyridin-4-amine: A mixture of 3-(4-chloropyridin-2-yl)-N-isopropyl-l- trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (40.0 mg, 0.0756 mmol), morpholine (13.0 mg, 0.151 mmol), Pd 2 (dba) 3 (7.0 mg, 0.0076 mmol), Xantphos (9.0 mg, 0.015 mmol), and Cs 2 C0 3 (49.0 mg, 0.151 mmol) in dioxane (1.0 mL) in a microwave tube was stirred at 160 °C for 120 min under microwave irradiation.
  • Step 3 Synthesis of N-isopropyl-3-(4-morpholinopyridin-2-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine: To a mixture of N-isopropyl-3-(4-morpholinopyridin-2-yl)-l-trityl-lH- pyrazolo[4,3-c]pyridin-4-amine (13.0 mg, 0.0224 mmol) in dichloromethane (5.0 mL) was added trifluoroacetic acid (1.0 mL) and triethylsilane (0.20 mL). The mixture was stirred under reflux for 1 hour.
  • Step 1 Synthesis of 3-(6-chloropyrimidin-4-yl)-N-isopropyl-l -trityl-lH-pyrazolo[4,3- c]pyridin-4-amine: To a microwave tube was added N-isopropyl-3-(trimethylstannyl)-l- trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (200 mg, 0.344 mmol), 4,6-dichloropyrimidine (102 mg, 0.688 mmol), LiCl (58.0 mg, 1.38 mmol), Cul (13.0 mg, 0.0688 mmol), Pd(PPh 3 )4 (40.0 mg, 0.0344 mmol) and THF (3.0 mL).
  • reaction mixture was heated at 100 °C for 1 hour under microwave irradiation. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (1 :2) to afford the title compound as a yellow solid (80 mg, 44%).
  • Step 2 Synthesis of N-isopropyl-3-(6-morpholinopyrimidin-4-yl)-l -trityl-lH- pyrazolo[4,3-c]pyridin-4-amine: A mixture of 3-(6-chloropyrimidin-4-yl)-N-isopropyl-l- trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (80.0 mg, 0.151 mmol), morpholine (26.0 mg, 0.302 mmol), and Cs 2 C0 3 (98.0 mg, 0.302 mmol) in i-PrOH (10 mL) was refiuxed at 120 °C for 2 h. After filtration, the filtrate was evaporated to afford the title compound as a yellow solid (80 mg, 92%).
  • Step 3 Synthesis of N-isopropyl-3-(6-mo holinopyrimidin-4-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine: To a mixture of N-isopropyl-3-(6-mo holinopyrimidin-4-yl)-l-trityl- lH-pyrazolo[4,3-c]pyridin-4-amine (80.0 mg, 0.138 mmol) in dichloromethane (5.0 mL) was added trifluoroacetic acid (1.0 mL) and triethylsilane (0.20 mL). The mixture was stirred under reflux for 1 hour.
  • Step 1 Synthesis of N-isopropyl-3-(4-morpholinopyrimidin-2-yl)-l-trityl-lH- pyrazolo[4,3-c]pyridin-4-amine: To a microwave tube was added N-isopropyl-3- (trimethylstannyl)-l-trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (200 mg, 0.344 mmol), 4- (2-bromopyrimidin-4-yl)mo holine (168 mg, 0.688 mmol), LiCl (58.0 mg, 1.38 mmol), Cul (13.0 mg, 0.0688 mmol), Pd(PPh 3 ) 4 (40.0 mg, 0.0344 mmol), and THF (3.0 mL).
  • reaction mixture was heated at 100°C for 1 hour under microwave irradiation. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (1 :2) to afford the title compound as a yellow solid (25 mg, 13%).
  • Step 2 Synthesis of N-isopropyl-3-(4-mo holinopyrimidin-2-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine: To a mixture of N-isopropyl-3-(4-mo holinopyrimidin-2-yl)-l-trityl- lH-pyrazolo[4,3-c]pyridin-4-amine (25 mg, 0.043 mmol) in dichloromethane (5.0 mL) was added trifluoroacetic acid (1.0 mL) and triethylsilane (0.20 mL). The mixture was stirred under reflux for 1 hour.
  • Step 1 Synthesis of 3-(6-chloropyrimidin-4-yl)-N-isopropyl-l-trityl-lH- pyrazolo[4,3-c]pyridine-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-isopropyl-3-(trimethylstannyl)-l-trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (100 mg, 0.172 mmol), 4,6-dichloropyrimidine (51.0 mg, 0.344 mmol), LiCl (29.0 mg, 0.688 mmol), Cul (7.0 mg, 0.034 mmol), Pd(PPh 3 ) 4 (20.0 mg, 0.0172 mmol), and THF (2 mL).
  • reaction mixture was heated at 100 °C for 1 hour under microwave irradiation. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (1/2) to afford the title compound as a yellow solid (72 mg, 79%).
  • Step 2 Synthesis of N-isopropyl-3-(pyrimidin-4-yl)-l-trityl-lH-pyrazolo[4,3- c]pyridin-4-amine: To a mixture of 3-(6-chloropyrimidin-4-yl)-N-isopropyl-l-trityl-lH- pyrazolo[4,3-c]pyridine-4-amine (72.0 mg, 0.136 mmol) and Pd/C (10 mg, 10% w/w) in methanol (10 ml) was added a drop of NH 4 OH. The mixture was stirred at 55 °C under 3 ⁇ 4 overnight. After cooling down to room temperature, the mixture was filtered via Celite.
  • Step 3 Synthesis of N-isopropyl-3-(pyrimidin-4-yl)-lH-pyrazolo[4,3-c]pyridin-4- amine: To a mixture of N-isopropyl-3-(pyrimidin-4-yl)-l-trityl-lH-pyrazolo[4,3-c]pyridin- 4-amine (65.0 mg, 0.131 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.0 mL) and triethylsilane (0.20 mL). The mixture was stirred under reflux for 1 hour.
  • Step 1 Synthesis of 4-chloro-3-iodo-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridine: To a solution of 4-chloro-3-iodo-lH-pyrazolo[4,3-c]pyridine (2.00 g, 7.17 mmol) in DMF (30 mL) was added KOH (0.800 g, 14.3 mmol) and l-(chloromethyl)-4- methoxybenzene (2.24 g, 14.3 mmol). The mixture was stirred at room temperature overnight.
  • Step 2 Synthesis of N-ethyl-3-iodo-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 4- chloro-3-iodo-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridine (2.00 g, 3.84 mmol), ethanamine (865 mg, 19.2 mmol), and methanol (10 mL).
  • reaction mixture was heated at 150 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM/petroleum ether/TEA (2/1/0.01) to afford the title compound as a white solid (1.5 g, 73 %).
  • Step 3 Synthesis of N-ethyl-l-(4-methoxybenzyl)-3-(trimethylstannyl)-lH- pyrazolo[4,3-c]-pyridin-4-amine: A reaction tube equipped with a magnetic stirrer was charged with N-ethyl-3-iodo- 1 -(4-methoxybenzyl)- lH-pyrazolo[4,3-c]pyridin-4-amine (750 mg, 1.84 mmol), 1,1,1,2,2,2-hexamethyldistannane (721 mg, 2.21 mmol), toluene (12 mL), and trans-Pd(PPh 3 )2Cl2 (39.0 mg, 0.0551 mmol).
  • reaction mixture was sealed and heated at 140 °C for 0.5 h. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure to afford the crude title compound, which was used in next step directly without further purification.
  • Step 4 Synthesis of N-ethyl-l-(4-methoxybenzyl)-3-(pyridin-2-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-ethyl-l-(4-methoxybenzyl)-3-(trimethylstannyl)-lH-pyrazolo[4,3- c]pyridin-4-amine (100 mg, 0.225 mmol), 2-bromopyridine (71 mg, 0.45 mmol), LiCl (38 mg, 0.9 mmol), Cul (9.0 mg, 0.045 mmol), Pd(PPh 3 ) 4 (26.0 mg, 0.0225 mmol), and THF (3 mL).
  • reaction mixture was heated at 100 °C for 1 hour under microwave irradiation. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (1 :2) to afford the title compound as a yellow solid (74 mg, 91%).
  • Step 5 Synthesis of N-ethyl-3-(pyridin-2-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-ethyl-l-(4- methoxybenzyl)-3-(pyridin-2-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (74.0 mg, 0.206 mmol) and TFA (3.0 mL) . The reaction mixture was heated at 120 °C for 2 h under microwave irradiation. After cooling down to room temperature, the mixture was concentrated under reduced pressure.
  • Step 1 Synthesis 3-(6-chloropyrimidin-4-yl)-N-ethyl-l-(4-methoxybenzyl)-lH- pyrazolo-[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-ethyl-l-(4-methoxybenzyl)-3-(trimethylstannyl)-lH-pyrazolo[4,3- c]pyridin-4-amine (100 mg, 0.225 mmol), 4,6-dichloropyrimidine (67 mg, 0.45 mmol), LiCl (38 mg, 0.90 mmol), Cul (9.0 mg, 0.045 mmol), Pd(PPh 3 ) 4 (26.0 mg, 0.0225 mmol), and THF (3 mL).
  • reaction mixture was heated at 100 °C for 1 hour under microwave irradiation. After cooling down to room temperature, the mixture was filtered via Celite. The filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (1 :2) to afford the title compound as a yellow solid (78 mg, 87%).
  • Step 2 Synthesis of N-ethyl-l-(4-methoxybenzyl)-3-(pyrimidin-4-yl)-lH- pyrazolo[4,3-c]-pyridin-4-amine: To a mixture of 3-(6-chloropyrimidin-4-yl)-N-isopropyl- l-trityl-lH-pyrazolo[4,3-c]-pyridine-4-amine (78.0 mg, 0.198 mmol) and Pd/C (10 mg, 10% w/w) in methanol (10 ml) was added a drop of NH 4 OH. The mixture was stirred at 55 °C under 3 ⁇ 4 overnight. After cooling down, the reaction mixture was filterred via Celite. The filtrate was concentrated to give the title compound as a yellow solid (65 mg, 91 >).
  • Step 3 Synthesis of N-ethyl-3-(pyrimidin-4-yl)-lH-pyrazolo[4,3-c]pyridin-4- amine: A microwave vial equipped with a magnetic stirrer was charged with N-ethyl-l-(4- methoxybenzyl)-3-(pyrimidin-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (65 mg, 0.18 mmol) and TFA (3 mL) . The reaction mixture was heated at 120 °C for 2 h under microwave irradiation. After cooling down to room temperature, the mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC to give the title compound as yellow solid (18 mg, 41%).
  • Step 1 Synthesis of 3-iodo-l-(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4-yl)- lH-pyrazolo[4,3-c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with 4-chloro-3-iodo-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridine (2.60 g, 6.50 mmol), tetrahydro-2H-pyran-4-amine (1.97 g, 19.5 mmol), n-BuOH (15 mL), and DIPEA (2.53 g, 19.5 mmol).
  • the reaction mixture was heated at 180 °C for 4 h under microwave irradiation. After cooling down, the solid was collected and washed with EtOH to afford the title compound as a white solid (2.35 g, 78%).
  • Step 2 Synthesis of l-(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4-yl)-3- (trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine:
  • a reaction tube equipped with a magnetic stirrer was charged with 3-iodo-l-(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4- yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (600 mg, 1.29 mmol), 1,1,1,2,2,2- hexamethyldistannane (507 mg, 1.55 mmol), toluene (12 mL), and trans-Pd(PPh 3 )2Cl2 (24.0 mg, 0.0314 mol).
  • reaction mixture was heated at 140 °C for 1 h. After cooling down, the reaction mixture was filterred. The filtrate was concentrated to give the title compound as a yellow solid which was used for the next step without further purification (850 mg).
  • Step 3 Synthesis of l-(4-methoxybenzyl)- N-(tetrahydro-2H-pyran-4-yl)-3 -(6- (trifluo romethyl) pyrimidin-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with l-(4-methoxybenzyl)-N-(tetrahydro- 2H-pyran-4-yl)-3-(trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (500 mg, 0.997 mmol), 4-bromo-6-(trifiuoromethyl)pyrimidine (271 mg, 1.19 mmol), LiCl (184 mg, 4.28 mmol), Cul (20.0 mg, 0.107 mmol), Pd(PPh 3 ) 4 (124 mg, 0.107 mmol), and THF (15 mL).
  • reaction mixture was heated at 100 °C for 1 h under microwave irradiation. After cooling down, the reaction mixture was filterred. The filtrate was concentrated and the residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (1 :2) to afford the title compound as a yellow solid (180 mg, 49% yield over two steps).
  • Step 4 Synthesis of N-(tetrahydro-2H-pyran-4-yl) -3-(4-(trifluoromethyl) pyridin- 2-yl) -lH-pyrazolo [4,3-c] pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with l-(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4-yl)-3-(6- (trifluoromethyl) pyrimidin-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (100 mg, 0.210 mmol) and TFA (10 mL).
  • Step 1 Synthesis of 3-iodo-N-isopropyl-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with 4- chloro-3-iodo-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridine (2.00 g, 3.84 mmol), propan-2-amine (1.10 g, 19.2 mmol), n-BuOH (0.860 g, 7.68 mmol), and DIPEA (10 mL).
  • the reaction mixture was heated at 170 °C for 2 h under microwave irradiation.
  • Step 2 Synthesis of N-isopropyl-l-(4-methoxybenzyl)-3-(trimethylstannyl)-lH- pyrazolo[4,3-c]pyridin-4-amine: A reaction tube equipped with a magnetic stirrer was charged with 3-iodo-N-isopropyl-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4- amine (750 mg, 1.78 mmol), 1 ,1 ,1 ,2,2,2-hexamethyldistannane (700 mg, 2.14 mmol), toluene (12 mL), and trans-Pd(PPri3)2Cl2 (32 mg, 0.045 mmol).
  • reaction mixture was sealed and heated at 110°C for 12 h. After cooling down, the mixture was filtered and the filtrate was concentrated under reduced pressure to give the crude title compound, which was used in the next step directly without further purification.
  • Step 3 Synthesis of N-isopropyl-l-(4-methoxybenzyl)-3-(6- (trifluoromethyl)pyrimidin-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-isopropyl-l-(4-methoxybenzyl)-3- (trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (310 mg, 0.670 mmol), 4-bromo-6- (trifluoromethyl)pyrimidine (120 mg, 0.530 mmol), LiCl (113 mg, 2.68 mmol), Cul (25.5 mg, 0.134 mmol), Pd(PPh 3 ) 4 (77 mg, 0.067 mmol), and THF (8 mL).
  • Step 4 Synthesis of N-isopropyl-3-(6-(trifluoromethyl)pyrimidin-4-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-isopropyl-1 -(4-methoxybenzyl)-3-(6-(trifluoromethyl)pyrimidin-4-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine (200 mg, 0.450 mmol) and TFA (5 mL). The reaction mixture was heated at 150 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • Step 1 Synthesis of 3-(4-chloropyridin-2-yl)-N-isopropyl-l-(4-methoxybenzyl)- lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-isopropyl-l-(4-methoxybenzyl)-3-(trimethylstannyl)-lH-pyrazolo[4,3- c]pyridin-4-amine (100 mg, 0.220 mmol), 2-bromo-4-chloropyridine (34 mg, 0.18 mmol), LiCl (37 mg, 0.88 mmol), Cul (8.4 mg, 0.044 mmol), Pd(PPh 3 ) 4 (26 mg, 0.022 mmol), and THF (4 mL).
  • Step 2 Synthesis of 3-(4-((3R,4R)-3,4-difluoropyrrolidin-l-yl)pyridin-2-yl)-N-iso- propyl-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A mixture of 3-(4- chloropyridin-2-yl)-N-isopropyl- 1 -(4-methoxybenzyl)- 1 H-pyrazolo [4,3 -c]pyridin-4-amine (200 mg, 0.490 mmol), (3R,4R)-3,4-difluoropyrrolidine hydrochloride (63 mg, 0.59 mmol), Pd 2 (dba) 3 (45 mg, 0.049 mmol), Xantphos (57 mg, 0.098 mmol), cesium carbonate (559 mg, 1.72 mmol), dioxane (10 mL) was degas
  • Step 3 Synthesis of 3-(4-((3R,4R)-3,4-difiuoropyrrolidin-l-yl)pyridin-2-yl)-N- isopropyl-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 3-(4-((3R,4R)-3,4-difluoropyrrolidin-l-yl)pyridin-2-yl)- N-isopropyl-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (95.0 mg, 0.199 mmol) and TFA (3 mL).
  • Step 1 Synthesis of N-isopropyl-l-(4-methoxybenzyl)-3-(4-(l-methyl-lH-pyrazol- 4-yl)pyridin-2-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 3-(4-chloropyridin-2-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (150 mg, 0.370 mmol), l-methyl-4- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (116 mg, 0.560 mmol), Pd(tBu 2 PC6H4NMe 2 )2Cl2 (26 mg, 0.037 mmol), Na 2 C0 3 (156 mg, 1.48 mmol), CH
  • reaction mixture was sealed and heated at 145 °C for 45 minutes under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM/MeOH (20:1) to give the title product as a yellow solid (80 mg, 48%).
  • Step 2 Synthesis of N-isopropyl-3-(4-(l-methyl-lH-pyrazol-4-yl)pyridin-2-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine: A mixture of N-isopropyl-l-(4-methoxybenzyl)-3-(4-(l- methyl-lH-pyrazol-4-yl)pyridin-2-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (80 mg, 0.18 mmol) and TFA (15 mL) was stirred at reflux overnight. After cooling down and concentration, the mixture was adjusted pH around 7 by saturated NaHC0 3 solution.
  • Step 1 Synthesis of 3-(4-(lH-pyrazol-3-yl)pyridin-2-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: To a solution of 3-(4-chloropyridin-
  • Step 3 Synthesis of N-isopropyl-3-(4-(l-methyl-lH-pyrazol-3-yl)pyridin-2-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine: A solution of N-isopropyl-l-(4-methoxybenzyl)-3-(4-(l- methyl-lH-pyrazol-3-yl)pyridin-2-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (72 mg, 0.16 mmol) in TFA (1 mL) was heated at 120 °C for 45 min under microwave irradiation.
  • Step 1 Synthesis of 3-(4-cyclohexenylpyridin-2-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 3-(4-chloropyridin-2-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (350 mg, 0.860 mmol), 2-(cyclohex- 2-enyl)-4,4,5,5-tetramethyl-l ,3,2-dioxaborolane (266 mg, 1.29 mmol), Pd(tBu 2 PC6H4NMe 2 )2Cl2 (61 mg, 0.086 mmol), Na 2 C0 3 (0.43 mL, 2M), and CH 3 CN (10 mL).
  • Step 2 Synthesis of 3-(4-cyclohexylpyridin-2-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A three-neck flask equipped with a magnetic stirrer was charged with 3-(4-cyclohexenylpyridin-2-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (60.0 mg, 0.132 mmol), Pd/C (30 mg), and MeOH (10 mL). The suspension was heated at 65 °C under H 2 for 12 h.
  • Step 3 Synthesis of 3-(4-cyclohexylpyridin-2-yl)-N-isopropyl-lH-pyrazolo[4,3- c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 3- (4-cyclohexylpyridin-2-yl)-N-isopropyl-l -(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin- 4-amine (44 mg, 0.096 mmol) and TFA (5 mL).
  • Step 1 Synthesis of 3-(6-chloropyrimidin-4-yl)-N-isopropyl-l-(4-methoxybenzyl)- lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-isopropyl-l -(4-methoxybenzyl)-3-(trimethylstannyl)-lH-pyrazolo[4,3- c]pyridin-4-amine (300 mg, 0.660 mmol), 4,6-dichloropyrimidine (195 mg, 1.32 mmol), LiCl (50 mg, 1.2 mmol), Cul (50 mg, 0.26 mmol), Pd(PPh 3 ) 4 (76 mg, 0.066 mmol), and THF (30 mL).
  • reaction mixture was heated at 100 °C for 30 min under microwave irradiation. After cooling down, the mixture was filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with petroleum ether/ethyl acetate (1 :2) to afford the title compound as a yellow solid (250 mg, 92%).
  • Step 2 Synthesis of 3-(6-cyclohexenylpyrimidin-4-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 3-(6-chloropyrimidin-4-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (200 mg, 0.490 mmol), 2-(cyclohex- 2-enyl)-4,4,5,5-tetramethyl-l ,3,2-dioxaborolane (144 mg, 0.740 mmol), Pd(tBu 2 PC 6 H 4 NMe2)2Cl2 (35 mg, 0.049 mmol), Na 2 C0 3 (0.5 mL, 2M), and CH 3 CN (6 mL).
  • Step 3 Synthesis of 3-(6-cyclohexylpyrimidin-4-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A three-neck flask equipped with a magnetic stirrer was charged with 3-(6-cyclohexenylpyrimidin-4-yl)-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (130 mg, 0.290 mmol), Pd/C (13 mg), and MeOH (10 mL). The suspension was heated at 65 °C under 3 ⁇ 4 for 2 h. After cooling down, the mixture was filtered. The filtrate was concentrated under reduced pressure to afford the title compound as yellow oil (50 mg, 38%).
  • Step 4 Synthesis of 3-(6-cyclohexylpyrimidin-4-yl)-N-isopropyl-lH-pyrazolo[4,3- c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 3- (6-cyclohexylpyrimidin-4-yl)-N-isopropyl-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-4-amine (50 mg, 0.1 1 mmol) and TFA (0.5 mL). The reaction mixture was heated at 150 °C for 30 min under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • Step 1 Synthesis of methyl 2-(4-(isopropylamino)-l-(4-methoxybenzyl)-lH- pyrazolo[4,3-c]pyridin-3-yl)-isonicotinate:
  • a microwave vial equipped with a magnetic stirrer was charged with N-isopropyl-l-(4-methoxybenzyl)-3-(trimethylstannyl)-lH- pyrazolo[4,3-c]pyridin-4-amine (2.4 g, 5.2 mmol), methyl 2-bromoisonicotinate (1.35 g, 6.25 mmol), LiCl (877 mg, 20.8 mmol), Cul (99.0 mg, 0.522 mole), Pd(PPh 3 ) 4 (201 mg, 0.174 mol), and THF (30 mL).
  • Step 2 Synthesis of 2-(4-(isopropylamino)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-3-yl)isonicotinic acid: To a mixture of methyl 2-(4-(isopropylamino)-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-3-yl)isonicotinate (704 mg, 1.63 mmol) and LiOH (321 mg, 7.16 mmol) in THF (20 mL) and methane (20 mL), was added H 2 0 (10 mL). The resulting mixture was stirred at room temperature for 4 h.
  • Step 3 Synthesis of 2-(4-(isopropylamino)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-3-yl)-N,N-dimethylisonicotinamide: A mixture of 2-(4-(isopropylamino)-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-3-yl)isonicotinic acid (200 mg, 0.480 mmol), dimethylamine hydrochloride (59 mg, 0.72 mmol), HATU (274 mg, 0.720 mmol), and Et 3 N (145 mg, 1.44 mmol) in DMF (3 mL) was stirred at room temperature overnight. After removal of the volatiles, the residue was purified by prep-HPLC to afford the title compound as a yellow solid (130 mg, 55%).
  • Step 4 Synthesis of 2-(4-(isopropylamino)-lH-pyrazolo[4,3-c]pyridin-3-yl)-N,N- dimethylisonicotin amide:
  • a microwave vial equipped with a magnetic stirrer was charged with 2-(4-(isopropylamino)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-3-yl)-N,N- dimethylisonicotinamide (130 mg, 0.293 mmol) and TFA (3 mL).
  • the reaction mixture was heated at 120 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • Step 1 Synthesis of 6-(4-(isopropylamino)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-3-yl)pyrimidine-4-carboxylic acid: To a solution of ethyl 6-(4-(isopropylamino)- l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-3-yl)pyrimidine-4-carboxylate (140 mg, 0.312 mmol) in MeOH (2 mL) was added lithium hydroxide solution (2 mL, 2 M in water). The mixture was stirred at room temperature for 2 h. After removal of the volatiles, the residue was purified by reverse phase Combi-flash to give the title compound as white solid (100 mg, 76%).
  • Step 2 Synthesis of 6-(4-(isopropylamino)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-3-yl)-N,N-dimethylpyrimidine-4-carboxamide: To a solution of 6-(4- (isopropylamino)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-3-yl)pyrimidine-4- carboxylic acid (100 mg, 0.240 mmol) in methylene chloride (20 mL) was added oxalyl chloride (10 mL) and DMF (0.5 mL). The mixture was stirred at room temperature for 2 hours.
  • the acid chloride was treated with dimethylamine hydrochloride (40 mg, 0.48 mmol). After being stirred for 30 minutes at room temperature, the resulting mixture was concentrated. The residue was washed with water, extracted with DCM. The organic layer was dried and concentrated in vacuo to afford the title compound as a white solid (100 mg, 90%).
  • Step 3 Synthesis of 6-(4-(isopropylamino)-lH-pyrazolo[4,3-c]pyridin-3-yl)-N,N- dimethylpyrimidine-4-carboxamide:
  • a microwave vial equipped with a magnetic stirrer was charged with 6-(4-(isopropylamino)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin- 3-yl)-N,N-dimethylpyrimidine-4-carboxamide (50 mg, 0.11 mmol) and TFA (2 mL).
  • the reaction mixture was heated at 120 °C for 30 min under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • Step 1 Synthesis of 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4-ylamino)- lH-pyrazolo[4,3-c]pyridin-3-yl)isonicotinic acid: A microwave vial equipped with a magnetic stirrer was charged with 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4- ylamino)-lH-pyrazolo[4,3-c]pyridin-3-yl)isonicotinonitrile (250 mg, 0.570 mmol), KOH (3.3 g, 50 mmol), and H 2 0 (16.5 mL).
  • reaction mixture was heated at 100 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford the title compound as a white solid (230 mg, 88%).
  • Step 2 Synthesis of 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4-ylamino)- lH-pyrazolo[4,3-c]pyridin-3-yl)-N,N-dimethylisonicotinamide:
  • 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4-ylamino)-lH- pyrazolo[4,3-c]pyridin-3-yl)isonicotinic acid 200 mg, 0.440 mmol
  • dimethylamine 72 mg, 0.88 mmol
  • HATU 333 mg, 0.880 mmol
  • DIPEA 0.5 mL
  • DMF 20 mL.
  • Step 3 Synthesis of N,N-dimethyl-2-(4-(tetrahydro-2H-pyran-4-ylamino)-lH- pyrazolo[4,3-c]pyridin-3-yl)isonicotinamide:
  • a microwave vial equipped with a magnetic stirrer was charged with 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4-ylamino)-lH- pyrazolo[4,3-c]pyridin-3-yl)-N,N-dimethylisonicotinamide (186 mg, 0.390 mmol) and TFA (20 mL).
  • Step 1 Synthesis of methyl 2-(l-(4-methoxybenzyl) -4-(tetrahydro-2H- pyran-4- ylamino)-lH-pyrazolo[4,3-c]pyridin-3-yl)isonicotinate:
  • a microwave vial equipped with a magnetic stirrer was charged with l-(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4-yl)-3- (trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (200 mg, 0.399 mmol), methyl 2- bromoisonicotinate (172 mg, 0.798 mmol), LiCl (67 mg, 1.6 mmol), Cul (7.6 mg, 0.040 mmol), Pd(PPh 3 )4 (46 mg, 0.040 mmol), and THF (10 mL).
  • Step 2 Synthesis of 2-(4-(tetrahydro-2H-pyran-4-ylamino)-lH-pyrazolo[4,3- c]pyridin-3-yl)isonicotinic acid:
  • a microwave vial equipped with a magnetic stirrer was charged with methyl 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4-ylamino)-lH- pyrazolo[4,3-c]pyridin-3-yl)isonicotinate (86.0 mg, 0.181 mmol) and TFA (12 mL).
  • the reaction mixture was heated at 150 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • the residue was purified by reverse-phase prep-HPLC to afford the title compound as a green solid (50 mg, 81%).
  • Step 3 Synthesis of azetidin-l-yl(2-(4-(tetrahydro-2H-pyran-4-ylamino)-lH- pyrazolo[4,3-c]pyridin-3-yl)pyridin-4-yl) methanone: To a 25-mL round bottom flash was charge with 2-(4-(tetrahydro-2H-pyran-4-ylamino)-lH-pyrazolo[4,3-c]pyridin-3- yl)isonicotinic acid (50 mg, 0.15 mmol), azetidine hydrochloride (28 mg, 0.30 mmol), HATU (96.5 mg, 0.254 mmol), DIPEA (0.5 mL), and DMF (3 mL).
  • Step 1 Synthesis of 3-(4-chloropyridin-2-yl)-l-(4-methoxybenzyl)-N-(tetrahydro- 2H-pyran-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with l-(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4-yl)-3- (trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (500 mg, 0.990 mmol), 2-bromo-4- chloropyridine (287 mg, 1.49 mmol), LiCl (166 mg, 3.96 mmol), Cul (19 mg, 0.099 mmol), Pd(PPh 3 ) 4 (114 mg, 0.0990 mmol), and THF (20 mL).
  • reaction mixture was heated at 100 °C for 1 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl acetate (3:1) to afford the title compound as a white solid (230 mg, 52%).
  • Step 2 Synthesis of 3-(4-ethoxypyridin-2-yl)-l-(4-methoxybenzyl)-N-(tetrahydro- 2H-pyran-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 3-(4-chloropyridin-2-yl)-l-(4-methoxybenzyl)-N- (tetrahydro-2H-pyran-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (200 mg, 0.445 mmol) , sodium ethanolate (61 mg, 0.89 mmol), and THF(20 ml).
  • reaction mixture was heated at 100 °C for 1 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by reverse-phase prep- HPLC to afford the title compound as a white solid (160 mg, 78%).
  • Step 3 Synthesis of 3-(4-ethoxypyridin-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with (3-(4-ethoxypyridin-2-yl)-l -(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4- yl)-lH-pyrazolo[4,3-c]pyridin-4-amine (160 mg, 0.350 mmol) and TFA (20 mL).
  • the reaction mixture was heated at 150 °C for 2 h under microwave irradiation.
  • Step 1 Synthesis of l-(4-methoxybenzyl)-3-(4-(2,2,6,6- tetrafluoromorpholino)pyridin-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with 3- (4-chloropyridin-2-yl)- 1 -(4-methoxybenzyl)-N-(tetrahydro-2H-pyran-4-yl)- 1 H- pyrazolo[4,3-c]pyridin-4-amine (150 mg, 0.330 mmol), Pd 2 (dba) 3 (30 mg, 0.033 mmol), Xantphos (382 mg, 0.660 mmol), Cs 2 C0 3 ( 216 mg, 0.660 mmol), 2,2,6,6- tetrafluoromorpholine (105 mg,
  • reaction mixture was heated at 140 °C for 1 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by reverse-phase prep- HPLC to afford the title compound as a white solid (86 mg, 46%).
  • Step 2 Synthesis of 3-(4-(2,2,6,6-tetrafluoromo holino)pyridin-2-yl)-N- (tetrahydro-2H-pyran-4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with l-(4-methoxybenzyl)-3-(4-(2,2,6,6- tetrafluoromoi holino)pyridin-2-yl)-N-(tetrahydro-2H-pyran-4-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine (86 mg, 0.15 mmol) and TFA (20 mL).
  • Step 1 Synthesis of ethyl 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4- ylamino)-lH-pyrazolo[4,3-c]pyridin-3-yl)-6-methylisonicotinate:
  • a microwave vial equipped with a magnetic stirrer was charged with l-(4-methoxybenzyl)-N-(tetrahydro- 2H-pyran-4-yl)-3-(trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (500 mg, 0.990 mmol), ethyl 2-bromo-6-methylisonicotinate (362 mg, 1.49 mmol), LiCl (166 mg, 3.96 mmol), Cul (19 mg, 0.099 mmol), Pd(PPh 3 ) 4 (1 14 mg, 0.0990 mmol), and THF (20 mL).
  • reaction mixture was heated at 100 °C for 1 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by prep-TLC eluting with petroleum ether/ethyl estate (2:1) to afford the title compound as a white solid (200 mg, 40%).
  • Step 2 Synthesis of 2-methyl-6-(4-(tetrahydro-2H-pyran-4-ylamino)-lH- pyrazolo[4,3-c]pyridin-3-yl)isonicotinic acid:
  • a microwave vial equipped with a magnetic stirrer was charged with ethyl 2-(l-(4-methoxybenzyl)-4-(tetrahydro-2H-pyran-4- ylamino)-lH-pyrazolo[4,3-c]pyridin-3-yl)-6-methylisonicotinate (200 mg, 0.390 mmol) and TFA (20 mL).
  • the reaction mixture was heated at 150 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford the title compound as a white solid (100 mg, 73%).
  • Step 3 Synthesis of N,N,2-trimethyl-6-(4-(tetrahydro-2H-pyran-4-ylamino)-lH- pyrazolo[4,3-c]pyridin-3-yl)isonicotinamide: To a 25-mL round bottom flash was charge with 2-methyl-6-(4-(tetrahydro-2H-pyran-4-ylamino)-lH-pyrazolo[4,3-c]pyridin-3- yl)isonicotinic acid (100 mg, 0.280 mmol), dimethylamine (36 mg, 0.45 mmol), HATU (167 mg, 0.450 mmol), DIPEA (0.50 mL), and DMF (20 mL).
  • Step 1 Synthesis of 3-iodo-l-(4-methoxybenzyl)-N-(2,2,2-trifluoroethyl)-lH- pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 4-chloro-3-iodo-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridine (1.0 g, 2.5 mmol), 2,2,2-trifluoroethanamine hydrochloride (1.0 g, 7.5 mmol), DIPEA (2.40 g, 18.6 mmol), and n-BuOH (10 ml).
  • reaction mixture was heated at 190 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM/petroleum ether/TEA (2: 1 : 0.01) to afford the title compound as a white solid (300 mg, 25%).
  • Step 2 Synthesis of l-(4-methoxybenzyl)-N-(2,2,2-trifiuoroethyl)-3- (trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A reaction tube equipped with a magnetic stirrer was charged with 3-iodo-l-(4-methoxybenzyl)-N-(2,2,2-trifiuoroethyl)- lH-pyrazolo[4,3-c]pyridin-4-amine (750 mg, 1.62 mmol), 1,1,1,2,2,2- hexamethyldistannane (637 mg, 1.95 mmol), toluene (12 mL), and trans-Pd(PPh 3 )2Cl2 (39 mg, 0.055 mmol).
  • reaction mixture was sealed and heated at 140 °C for 0.5 h. After cooling down, the mixture was filtered. The filtrate was concentrated under reduced pressure to give the title compound, which was used in the next step without further purification.
  • Step 3 Synthesis of l-(4-methoxybenzyl)-3-(pyridin-2-yl)-N-(2,2,2-trifiuoroethyl)- lH-pyrazolo[4,3-c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with N-ethyl-l-(4-methoxybenzyl)-3-(trimethylstannyl)-lH-pyrazolo[4,3- c]pyridin-4-amine (100 mg, 0.200 mmol), 2-bromopyridine (63 mg, 0.40 mmol), LiCl (34 mg, 0.80 mmol), Cul (8.0 mg, 0.040 mmol), Pd(PPh 3 ) 4 (24 mg, 0.020 mmol), and THF (3 mL).
  • Step 4 Synthesis of 3-(pyridin-2-yl)-N-(2,2,2-trifiuoroethyl)-lH-pyrazolo[4,3- c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with 1- (4-methoxybenzyl)-3-(pyridin-2-yl)-N-(2,2,2-trifluoroethyl)-lH-pyrazolo[4,3-c]pyridin-4- amine (76.0 mg, 0.184 mole) and TFA (3 mL) . The reaction mixture was heated at 120 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • Step 1 Synthesis of N-(4,4-difluorocyclohexyl)-3-iodo-l-(4-methoxybenzyl)-lH- pyrazolo[4,3-c]pyridin-4-amine:
  • a microwave vial equipped with a magnetic stirrer was charged with 4-chloro-3-iodo-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridine (400 mg, 1.00 mmol), 4,4-difluorocyclohexanamine hydrochloride (342 mg, 2.00 mmol), DIPEA (336 mg, 3.00 mmol), and n-BuOH (10 mL).
  • reaction mixture was heated at 170 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with DCM/petroleum ether/TEA (2:1 :0.01) to afford the title compound as a grey solid (310 mg, 62%).
  • Step 2 Synthesis of N-(4,4-difluorocyclohexyl)-l-(4-methoxybenzyl)-3- (trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A reaction tube equipped with a magnetic stirrer was charged with N-(4,4-difluorocyclohexyl)-3-iodo-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (240 mg, 0.482 mmol), 1,1,1,2,2,2- hexamethyldistannane (315 mg, 0.946 mmol), toluene (12 mL), and trans-Pd(PPri3)2Cl2 (8.5 mg, 0.012 mmol).
  • Step 3 Synthesis of N-(4,4-difiuorocyclohexyl)-l-(4-methoxybenzyl)-3-(pyridin-2- yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-(4,4-difluorocyclohexyl)-l-(4-methoxybenzyl)-3-(trimethylstannyl)- lH-pyrazolo[4,3-c]pyridin-4-amine (300 mg, 0.561 mmol), 2-bromopyridine (177 mg, 1.12 mmol), LiCl (94.0 mg, 2.24 mmol), Cul (11 mg, 0.056 mmol), Pd(PPh 3 ) 4 (63 mg, 0.056 mmol), and THF (3 mL).
  • Step 4 Synthesis of N-(4,4-difluorocyclohexyl)-3-(pyridin-2-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-(4,4-difluorocyclohexyl)-l-(4-methoxybenzyl)-3-(pyridin-2-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine (80.0 mg, 0.178 mmol) and TFA (3 mL). The reaction mixture was heated at 120 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • Step 1 Synthesis of 3-(6-chloropyrimidin-4-yl)-N-(4,4-difiuorocyclohexyl)-l-(4- methoxybenz-y-l)-lH-pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-(4,4-difluorocyclohexyl)-l-(4-methoxybenzyl)-3- (trimethylstannyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (300 mg, 0.561 mmol), 4,6- dichloropyrimidine (166 mg, 1.12 mmol), LiCl (94.0 mg, 2.24 mmol), Cul (1 1 mg, 0.056 mmol), Pd(PPh 3 )4 (63 mg, 0.056 mmol), and THF (3 mL).
  • Step 2 Synthesis of N-(4,4-difluorocyclohexyl)-l-(4-methoxybenzyl)-3-(pyrimidin- 4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: A mixture of 3-(6-chloropyrimidin-4-yl)-N-(4,4- difluorocyclohexyl)-l-(4-methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (110 mg, 0.227 mmol) and Pd/C (20 mg ) in methanol (20 mL) was stirred at 50 °C under 3 ⁇ 4 for 10 h. The insoluble was filtered off and the filtrate was concentrated under reduced pressure to give the title compound as white oil (90 mg, 88%).
  • Step 3 Synthesis of N-(4,4-difluorocyclohexyl)-3-(pyrimidin-4-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine: A microwave vial equipped with a magnetic stirrer was charged with N-(4,4-difluorocyclohexyl)-l-(4-methoxybenzyl)-3-(pyrimidin-4-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine (90 mg, 0.20 mmol) and TFA (3 mL). The reaction mixture was heated at 120 °C for 2 h under microwave irradiation. After cooling down, the mixture was concentrated under reduced pressure.
  • lodomethane (0.011 mL, 0.180 mmol) was added, the vial was capped and the reaction was heated to 90 °C for 2 h.
  • the reaction mixture was diluted with DCM and water and the layers were separated.
  • the aqueous layer was extracted with DCM (2x), the combined organics were dried over sodium sulfate, filtered and concentrated in vacuo.
  • the crude residue was purified by column chromatography (0-100% EtOAc in DCM). Recovered two regioisomers (20.0 mg, 40%>; 12.9 mg, 26%>).
  • Step 1 Synthesis of 3-(5-ethyl-lH-pyrazol-3-yl)-N-(tetrahydro-2H-pyran-4-yl)-l- trityl-lH-pyrazolo[4,3-c]pyridin-4-amine: To a microwave vial was added l-(4- ((tetrahydro-2H-pyran-4-yl)amino)- 1 -trityl- 1 H-pyrazolo [4,3 -c]pyridin-3-yl)pent- 1 -en-3- one (0.114 g, 0.210 mmol) followed by N,N-dimethylformamide (2.43 mL, 31.4 mmol).
  • Step 2 Synthesis of 3-(5-ethyl-l-methyl-lH-pyrazol-3-yl)-N-(tetrahydro-2H-pyran- 4-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: To a solution of 3-(5-ethyl-lH-pyrazol-3-yl)-N- (tetrahydro-2H-pyran-4-yl)-l-trityl-lH-pyrazolo[4,3-c]pyridin-4-amine (0.114 g, 0.206 mmol) in N,N-dimethylformamide (3.0 mL, 39 mmol) at rt was added cesium carbonate (0.100 g, 0.308 mmol) and methyl iodide (0.0192 mL, 0.308 mmol).
  • Step 1 Synthesis of 4-chloro-3-(l-isopropyl-lH-imidazol-4-yl)-l-trityl-lH- pyrazolo[4,3-c]pyridine: To a microwave vial was added 4-chloro-3-iodo-l-trityl-lH- pyrazolo[4,3-c]pyridine (0.44 g, 0.00085 mol), l-isopropyl-4-(tributylstannyl)-lH- imidazole (0.338 g, 0.847 mmol), [1 ,1'- bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.069 g, 0.085 mmol), copper(I) thiophene-2-carboxylate (0.242 g, 1.27 mmol), cesium fluoride (0.26 g, 1.70 mmol).
  • Step 2 Synthesis of 3-(l-isopropyl-lH-imidazol-4-yl)-N-(tetrahydro-2H-pyran-4- yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: To a microwave vial was added crude 4-chloro-3- (l-isopropyl-lH-imidazol-4-yl)-l-trityl-lH-pyrazolo[4,3-c]pyridine (0.064 g, 0.13 mmol), 4-aminotetrahydropyran (0.0154 mL, 0.152 mmol), BrettPhos palladium pre -catalyst (0.0101 g, 0.0127 mmol), BrettPhos (0.00682 g, 0.0127 mmol) and sodium feri-butoxide (0.0293 g, 0.305 mmol).
  • the vial was purged with nitrogen and 1 ,4-dioxane (1.4 mL, 18 mmol) was added. Nitrogen was then bubbled through the reaction mixture for 5 mins then the reaction mixture was heated in an oil bath to 110 °C overnight. The reaction was then filtered through Celite, eluting with DCM, concentrated in vacuo and purified by column chromatography (0-10% DCM in MeOH). The combined product fractions were concentrated in vacuo and dissolved in dichloromethane (1.8 mL, 28 mmol).
  • Triethylsilane (0.0811 mL, 0.508 mmol) and trifluoro acetic acid (1.8 mL, 23 mmol) were added and the reaction was stirred at room temperature for 15 mins. The reaction was concentrated in vacuo and the crude residue was purified by reverse-phase HPLC to give the title compound (4.6 mg, 11% over two steps).
  • Step 1 Synthesis of 3-ethynyl-N-isopropyl-l-(4-methoxybenzyl)-lH-pyrazolo[4,3- c]pyridin-4-amine: To a microwave vial was added 3-iodo-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (0.350 g, 0.829 mmol), bis(triphenylphosphine)palladium(II) chloride (0.0451 g, 0.0643 mmol), copper(I) iodide (0.0122 g, 0.0643 mmol).
  • the vial was purged with nitrogen and acetonitrile (2.2 mL, 43 mmol), triethylamine (0.470 mL, 3.37 mol), and (trimethylsilyl)acetylene (0.136 mL, 0.964 mmol) were added sequentially.
  • the vial was sealed and stirred at rt for 1 h.
  • the reaction mixture was filtered through Celite, eluting with DCM and concentrated in vacuo.
  • the residue was purified by column chromatography (0-5% EtOAc in DCM). The combined product fractions were concentrated in vacuo and dissolved in methanol (60.2 mL, 149 mmol).
  • Step 2 Synthesis of N-isopropyl-3-(l-isopropyl-lH-l,2,3-triazol-4-yl)-lH- pyrazolo[4,3-c]pyridin-4-amine: To a vial was added 3-ethynyl-N-isopropyl-l-(4- methoxybenzyl)-lH-pyrazolo[4,3-c]pyridin-4-amine (0.0907 g, 0.283 mmol), sodium azide (0.0221 g, 0.340 mmol), and copper(I) iodide (0.00539 g, 0.0283 mmol).
  • Trifluoromethanesulfonic acid (0.15 mL, 1.7 mmol) was added and the reaction mixture was stirred for 1 h at room temp.
  • the reaction mixture was diluted with DCM and water and made basic by slow addition of sat. aq. sodium bicarbonate.
  • the organic layer was separated and the aqueous layer was extracted with CHCI 3 : ; PrOH (5:1) (3x).
  • the combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo.
  • the crude residue was purified by reverse-phase HPLC to give the title compound (25.9 mg, 32% over two steps). !
  • Step 1 Synthesis of N-isopropyl-l-(4-methoxybenzyl)-3-(4-(trifluoromethyl)-lH- pyrazol-l-yl)-lH-pyrazolo[4,3-c]pyridin-4-amine: To a vial was added 3-iodo-N- isopropyl-l-[(4-methoxyphenyl)methyl]pyrazolo[4,3-c]pyridin-4-amine (100 mg, 0.237 mmol), 4-(trifiuoromethyl)-lH-pyrazole (30.6 mg, 0.225 mmol), copper(I) iodide (11.3 mg, 0.059 mmol), potassium carbonate (57.8 mg, 0.414 mmol).
  • the vial was purged with nitrogen and toluene (0.47 mL, 4.45 mmol) and ira «s-N,N'-dimethylcyclohexane-l,2- diamine, (0.02 mL, 0.12 mmol) were added.
  • the vial was sealed and heated to 110 °C for 24 h.
  • the reaction mixture was filtered through Celite, eluting with DCM and concentrated in vacuo.
  • the crude residue was purified by column chromatography (0-20% EtOAc in DCM) to give the title compound which was used directly in the next step.
  • Step 2 Synthesis of N-isopropyl-l-[(4-methoxyphenyl)methyl]-3-[4- (trifluoromethyl)pyrazol-l-yl]pyrazolo[4,3-c]pyridin-4-amine: To a solution of N- isopropyl-l-(4-methoxybenzyl)-3-(4-(trifluoromethyl)-lH-pyrazol-l-yl)-lH-pyrazolo[4,3- c]pyridin-4-amine (15.2 mg, 0.035 mmol) in dichloromethane (1.0 mL; 16 mmol) was added trifiuoromethanesulfonic acid (0.032 mL, 0.35 mmol).

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Abstract

L'invention porte sur un composé de formule (1A) ou (1B), ou un sel pharmaceutiquement acceptable de celui-ci, dans laquelle formule R1 représente un groupe choisi parmi un groupe alkyle, un groupe hétérocycloalkyle monocyclique, un groupe hétérocycloalkyle bicyclique et un groupe cycloalkyle, chacun de ceux-ci étant éventuellement substitué par un ou plusieurs groupes choisis parmi un groupe alkyle, un atome d'halogène et un groupe cycloalkyle ; pour la formule (1A) : un, deux ou trois groupes parmi X1, X2, X3 et X4 représentent N et les autres représentent chacun indépendamment CR2 ; ou pour la formule (1B) : X4 représente C ou N ; et un ou deux des groupes X1, X2 et X3 sont indépendamment choisis entre N et NR8 et les autres représentent chacun indépendamment CR2 ; de façon à ce que X1, X2, X3, X4 et N forment un groupe hétéroaryle ; chaque R2 est indépendamment choisi parmi H, un groupe alkyle, CN, un atome d'halogène, un groupe hétéroaryle, un groupe hétérocycloalkyle, un groupe cycloalkyle, OR4, CONR5R6 et SO2R7, lesdits groupes alkyle, hétéroaryle, hétérocycloalkyle et cycloalkyle étant chacun éventuellement en outre substitués par un ou plusieurs groupes choisis parmi un groupe alkyle, un atome d'halogène et OR9 ; chaque R8 est indépendamment choisi entre H et un groupe alkyle, ledit groupe alkyle étant éventuellement en outre substitué par un ou plusieurs groupes choisis parmi CN, un atome d'halogène, un groupe hétéroaryle, un groupe hétérocycloalkyle, un groupe cycloalkyle, OR4, CONR5R6 et SO2R7 ; R4, R5, R6, R7 et R9 sont chacun indépendamment choisis entre H et un groupe alkyle ; ou R5 et R6 conjointement avec l'atome d'azote auquel ils sont attachés sont liés pour former un groupe cyclique qui comprend en outre éventuellement un ou plusieurs hétéroatomes choisis parmi O, N et S ; à condition que, lorsque le composé est représenté par la formule (1B), le composé soit autre que la 3-(5-isopropyl-4H-1,2,4-triazol-3-yl)-N-(tétrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine ou la 3-(5-cyclopropyl-4H-1,2,4-triazol-3-yl)-N-(tétrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine. L'invention dans d'autres aspects porte sur des compositions pharmaceutiques, sur des utilisations thérapeutiques et sur un procédé pour la préparation de composés de formule (1A) et (1B).
PCT/EP2013/055878 2012-03-21 2013-03-21 Pyrazolopyridines pour le traitement de la maladie de parkinson WO2013139882A1 (fr)

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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015113452A1 (fr) * 2014-01-29 2015-08-06 Glaxosmithkline Intellectual Property Development Limited Composés
WO2015113451A1 (fr) * 2014-01-29 2015-08-06 Glaxosmithkline Intellectual Property Development Limited Composés
US9156845B2 (en) 2012-06-29 2015-10-13 Pfizer Inc. 4-(substituted amino)-7H-pyrrolo[2,3-d] pyrimidines as LRRK2 inhibitors
US9695171B2 (en) 2013-12-17 2017-07-04 Pfizer Inc. 3,4-disubstituted-1 H-pyrrolo[2,3-b]pyridines and 4,5-disubstituted-7H-pyrrolo[2,3-c]pyridazines as LRRK2 inhibitors
WO2017215600A1 (fr) * 2016-06-15 2017-12-21 Suzhou Yabao Pharmaceutical R&D Co., Ltd. Composés héterocycliques tricycliques substitués et leur utilisation
US10039753B2 (en) 2015-09-14 2018-08-07 Pfizer Inc. Imidazo[4,5-c]quinoline and imidazo[4,5-c][1,5]naphthyridine derivatives as LRRK2 inhibitors
WO2020021149A1 (fr) 2018-07-25 2020-01-30 Consejo Superior De Investigaciones Científicas Procédé pour déposer des éléments sur un substrat d'intérêt et dispositif
WO2020039110A1 (fr) 2018-08-24 2020-02-27 Consejo Superior De Investigaciones Científicas Composés inhibiteurs de lrrk2 et leur utilisation pour le traitement de maladies neurodégénératives
CN111615514A (zh) * 2018-01-18 2020-09-01 奥瑞生物药品公司 作为ret激酶抑制剂的取代的吡唑并[4,3-c]吡啶化合物
US10858359B2 (en) 2016-06-07 2020-12-08 Jacobio Pharmaceuticals Co., Ltd. Heterocyclic ring derivatives useful as SHP2 inhibitors
US10988466B2 (en) 2017-03-23 2021-04-27 Jacobio Pharmaceuticals Co., Ltd. Heterocyclic derivatives useful as SHP2 inhibitors
US11524963B2 (en) 2018-01-18 2022-12-13 Array Biopharma Inc. Substituted pyrazolo[3,4-d]pyrimidines as RET kinase inhibitors
WO2023076404A1 (fr) 2021-10-27 2023-05-04 Aria Pharmaceuticals, Inc. Méthodes de traitement de lupus érythémateux disséminé
WO2023160652A1 (fr) * 2022-02-25 2023-08-31 苏州亚宝药物研发有限公司 Composé cyclique fusionné substitué, composition pharmaceutique associée, procédé de préparation et utilisations associés

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984003564A1 (fr) 1983-03-08 1984-09-13 Commw Serum Lab Commission Procede de determination de sequences d'acides amines antigeniquement actives
WO2010106333A1 (fr) 2009-03-19 2010-09-23 Medical Research Council Technology Composés
WO2011141756A1 (fr) 2010-05-14 2011-11-17 Medical Research Council Technology Pyrazolopyridines en tant qu'inhibiteurs de la kinase lrrk2
WO2012038743A1 (fr) * 2010-09-22 2012-03-29 Medical Research Council Technology Pyrazolopyridines comme inhibiteurs de la kinase lrrk2

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984003564A1 (fr) 1983-03-08 1984-09-13 Commw Serum Lab Commission Procede de determination de sequences d'acides amines antigeniquement actives
WO2010106333A1 (fr) 2009-03-19 2010-09-23 Medical Research Council Technology Composés
WO2011141756A1 (fr) 2010-05-14 2011-11-17 Medical Research Council Technology Pyrazolopyridines en tant qu'inhibiteurs de la kinase lrrk2
WO2012038743A1 (fr) * 2010-09-22 2012-03-29 Medical Research Council Technology Pyrazolopyridines comme inhibiteurs de la kinase lrrk2

Non-Patent Citations (22)

* Cited by examiner, † Cited by third party
Title
"Advanced Organic Chemistry", 1985, JOHN WILEY AND SONS
"Handbook of Pharmaceutical Excipients", 1994
"Remington's Pharmaceutical Sciences", 1985, MACK PUBLISHING CO.
BERGE ET AL., J PHARM SCI, vol. 66, 1977, pages 1 - 19
BOSGRAAF, L.; RUSSCHER, H.; SMITH, J. L.; WESSELS, D.; SOLL, D. R.; VAN HAASTERT, P. J.: "A novel cGMP signalling pathway mediating myosin phosphorylation and chemotaxis in Dictyostelium", EMBO J., vol. 21, 2002, pages 4560 - 4570
BOSGRAAF, L.; VAN HAASTERT, P. J.: "Roc, a Ras/GTPase domain in complex proteins", BIOCHIM BIOPHYS ACTA, vol. 1643, 2003, pages 5 - 10, XP004477079, DOI: doi:10.1016/j.bbamcr.2003.08.008
BRETSCHER, A.; EDWARDS, K.; FEHON, R. G.: "ERM proteins and merlin: integrators at the cell cortex", NAT REV MOL CELL BIOL., vol. 3, 2002, pages 586 - 599
COHEN, P.; KNEBEL, A.: "KESTREL: a powerful method for identifying the physiological substrates of protein kinases", BIOCHEM J., vol. 393, 2006, pages 1 - 6
FARRER, M.; STONE, J.; MATA, I. F.; LINCOLN, S.; KACHERGUS, J.; HULIHAN, M.; STRAIN, K. J.; MARAGANORE, D. M.: "LRRK2 mutations in Parkinson disease", NEUROLOGY, vol. 65, 2005, pages 738 - 740
FINGL: "The Pharmacological Basis of Therapeutics", 1975, pages: 1
GOLDBERG, J. M.; BOSGRAAF, L.; VAN HAASTERT, P. J.; SMITH, J. L.: "Identification of four candidate cGMP targets in Dictyostelium", PROC NATL ACAD SCI USA., vol. 99, 2002, pages 6749 - 6754
GREGGIO, E.; JAIN, S.; KINGSBURY, A.; BANDOPADHYAY, R.; LEWIS, P.; KAGANOVICH, A.; VAN DER BRUG, M. P.; BEILINA, A.; BLACKINTON, J: "Kinase activity is required for the toxic effects of mutant LRRK2/dardarin", NEUROBIOL DIS., vol. 23, 2006, pages 329 - 341
JALEEL, M.; NICHOLS, R. J.; DEAK, M.; CAMPBELL, D. G.; GILLARDON, F.; KNEBEL, A.; ALESSI, D. R.: "LRRK2 phosphorylates moesin at threonine-558: characterization of how Parkinson's disease mutants affect kinase activity", BIOCHEM J., vol. 405, 2007, pages 307 - 317, XP002494862
MANNING, G.; WHYTE, D. B.; MARTINEZ, R.; HUNTER, T.; SUDARSANAM, S.: "The protein kinase complement of the human genome", SCIENCE, vol. 298, 2002, pages 1912 - 1934, XP002422776, DOI: doi:10.1126/science.1075762
MARIN, 1.: "The Parkinson disease gene LRRK2: evolutionary and structural insights.", MOL BIOL EVOL., vol. 23, 2006, pages 2423 - 2433
MATA, 1. F.; WEDEMEYER, W. J.; FARRER, M. J.; TAYLOR, J. P.; GALLO, K. A.: "LRRK2 in Parkinson's disease: protein domains and functional insights", TRENDS NEUROSCI., vol. 29, 2006, pages 286 - 293, XP025031251, DOI: doi:10.1016/j.tins.2006.03.006
NICHOLS, R. J.; DZAMKO, N.; HUTTI, J. E.; CANTLEY, L. C.; DEAK, M.; MORAN, J.; BAMBOROUGH, P.; REITH, A. D.; ALESSI, D. R.: "Substrate specificity and inhibitors of LRRK2, a protein kinase mutated in Parkinson's disease", BIOCHEM J., vol. 424, 2009, pages 47 - 60, XP002654035, DOI: doi:10.1042/BJ20091035
PAISAN-RUIZ, C.; JAIN, S.; EVANS, E. W.; GILKS, W. P.; SIMON, J.; VAN DER BRUG, M.; LOPEZ DE MUNAIN, A.; APARICIO, S.; GIL, A. M.;: "Cloning of the gene containing mutations that cause PARKB-linked Parkinson's disease", NEURON, vol. 44, 2004, pages 595 - 600, XP002371965, DOI: doi:10.1016/j.neuron.2004.10.023
POLESELLO, C.; PAYRE, F.: "Small is beautiful: what flies tell us about ERM protein function in development.", TRENDS CELL BIOL., vol. 14, 2004, pages 294 - 302
TAYLOR, J. P.; MATA, 1. F.; FARRER, M. J.: "LRRK2: a common pathway for parkinsonism, pathogenesis and prevention?", TRENDS MOL MED., vol. 12, 2006, pages 76 - 82, XP028058701, DOI: doi:10.1016/j.molmed.2005.12.004
WEST, A. B.; MOORE, D. J.; BISKUP, S.; BUGAYENKO, A.; SMITH, W. W.; ROSS, C. A.; DAWSON, V. L.; DAWSON, T. M.: "Parkinson's disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity", PROC NATL ACAD SCI USA., vol. 102, 2005, pages 16842 - 16847, XP002437140, DOI: doi:10.1073/pnas.0507360102
ZABETIAN, C. P.; SAMII, A.; MOSLEY, A. D.; ROBERTS, J. W.; LEIS, B. C.; YEAROUT, D.; RASKIND, W. H.; GRIFFITH, A.: "A clinic-based study of the LRRK2 gene in Parkinson disease yields new mutations.", NEUROLOGY, vol. 65, 2005, pages 741 - 744

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US9695171B2 (en) 2013-12-17 2017-07-04 Pfizer Inc. 3,4-disubstituted-1 H-pyrrolo[2,3-b]pyridines and 4,5-disubstituted-7H-pyrrolo[2,3-c]pyridazines as LRRK2 inhibitors
CN105940004A (zh) * 2014-01-29 2016-09-14 葛兰素史密斯克莱知识产权发展有限公司 化合物
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US10618901B2 (en) 2014-01-29 2020-04-14 Glaxosmithkline Intellectual Property Development Limited LRRK2 inhibitors for the treatment of Parkinson's disease
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US9815841B2 (en) 2014-01-29 2017-11-14 Glaxosmithkline Intellectual Property Development Limited Compounds
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US10039753B2 (en) 2015-09-14 2018-08-07 Pfizer Inc. Imidazo[4,5-c]quinoline and imidazo[4,5-c][1,5]naphthyridine derivatives as LRRK2 inhibitors
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US10918645B2 (en) 2016-06-15 2021-02-16 Lifearc Substituted tricyclic heterocyclic compounds and use thereof
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US10988466B2 (en) 2017-03-23 2021-04-27 Jacobio Pharmaceuticals Co., Ltd. Heterocyclic derivatives useful as SHP2 inhibitors
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US11472802B2 (en) 2018-01-18 2022-10-18 Array Biopharma Inc. Substituted pyrazolyl[4,3-c]pyridine compounds as RET kinase inhibitors
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WO2020021149A1 (fr) 2018-07-25 2020-01-30 Consejo Superior De Investigaciones Científicas Procédé pour déposer des éléments sur un substrat d'intérêt et dispositif
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