WO2010118207A1 - Pyrazolo [1, 5-a] pyrimidine derivatives as mtor inhibitors - Google Patents

Pyrazolo [1, 5-a] pyrimidine derivatives as mtor inhibitors Download PDF

Info

Publication number
WO2010118207A1
WO2010118207A1 PCT/US2010/030350 US2010030350W WO2010118207A1 WO 2010118207 A1 WO2010118207 A1 WO 2010118207A1 US 2010030350 W US2010030350 W US 2010030350W WO 2010118207 A1 WO2010118207 A1 WO 2010118207A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
substituted
heteroaryl
group
methyl
Prior art date
Application number
PCT/US2010/030350
Other languages
French (fr)
Inventor
Yongqi Deng
Binyuan Sun
Hongbo Zeng
Matthew Richards
Gerald W. Shipps, Jr.
Cliff C. Cheng
Yinyan Zhao
Andrew Mcriner
Zhaoyang Meng
Yang Nan
Mehul F. Patel
Iwona E. Wrona
Panduranga Adulla Reddy
Brian M. Eklov
Shuyi Tang
Duan Liu
Amit K. Mandal
Lianyun Zhao
M. Arshad Siddiqui
Original Assignee
Schering Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Schering Corporation filed Critical Schering Corporation
Priority to EP10715010.4A priority Critical patent/EP2417138B1/en
Priority to US13/263,193 priority patent/US8591943B2/en
Publication of WO2010118207A1 publication Critical patent/WO2010118207A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • This invention is directed to pyrazolo pyrimidine derivatives as inhibitors of mammalian Target Of Rapamycin (mTOR) kinase, which are also known as FRAP, RAFT, RAPT or SEP and as a result have a role in controlling cell cycle progression and mitogenic signals involved in cell growth and proliferation and as a result are useful in the treatment of cancer.
  • mTOR mammalian Target Of Rapamycin
  • rapamycin The mammalian target of rapamycin (mTOR) is a central regulator of cell growth and proliferation and plays a gate keeper role in the control of cell cycle progression and mediates mitogenic signals from P13K/AKT through to the downstream targets S6K1 and 4E-BP1 and to Ser 473 on AKT.
  • mTORCI raptor-mTOR complex
  • mTORC2 rapamycin-sensitive complex
  • mTORC2 a rapamycin-insensitive complex that signals to AKT.
  • rapamycin partially inhibits mTOR function through mTORCI . It has been found that mTORC2 is involved in the regulation of cell survival and actin cytoskeletal organization in a rapamycin-independent manner, and inhibition of mTOR through inhibition of mTORCI and mTORC2 is probably important for antitumor activity and better efficacy.
  • WO 2007/087395 describes unsaturated mTOR inhibitors useful in treatment of cancer.
  • WO-2008/012326 describes 2,4-substituted quinozolines as lipid kinase inhibitors useful in treatment of P13K -related diseases, such as proliferative diseases, inflammatory diseases, obstructive airways disorder and transplant related diseases.
  • WO 2006/090169 describes 2,4-diamineo-pyrido-pyrmidine derivatives and their use as mTOR inhibitors.
  • WO 2007/066099 describes pyrimidine derivatives useful as mTOR kinase inhibitors for anticancer and various other therapeutic treatments involving mTOR kinase.
  • WO 2007044813 describes pyridopyrimidinone inhibitors of PI3Ka protein kinase for anticancer or other PI3Ka protein kinase related diseases.
  • US 2002/0041880 describes pyrazolo[1 ,5 a]pyrimidin-7-yl derivatives to inhibit kinase insert domain-containing receptor to block angiogenesis.
  • WO 1998/003510 describes pyrazolo[1 ,5 a]pyrimidin-7-yl derivatives to treat corticotrophin releasing factor dependent diseases.
  • WO 2003/091256 describes pyrazolo[1 ,5-a]pyrimidine derivatives to treat NAD(P)H oxidase dependent diseases.
  • WO 2007/044449, WO 2005/077954, WO 2004/022560 and WO 2004/022561 describe pyrazolopyrimidine derivatives as cyclin-dependent kinase dependent diseases.
  • WO 2004/106341 describes pyrazolopyrimidine derivatives as fungicides There is need in the art for small molecule inhibitors of mTOR kinase that block signaling through mTORCI and mT0RC2 as a potential anticancer treatment or a treatment for other cell proliferative disorders.
  • the present invention discloses novel compounds having mTOR inhibitory activity, methods of preparing such compounds, pharmaceutical compositions comprising one or more of such compounds, methods of treatment or prevention of one or more diseases associated with mTOR by administering one or more of such compounds or pharmaceutical compositions, the compound being represented by the general Formula I:
  • R 1 is independently selected from the group consisting of heterocycloalkyl, heterocycloalkylalkyl, spiroheterocycloalkyl, heterocyclenyl, -NR 3 R 4 , cycloalkyl, heteroaryl, aryl, alkyl, alkynyl, heterocyclenylalkyl, cycloalkylalkyl, heteroarylalkyl, heteroarylalkynyl, spiroheterocycloalkylalkyl, -N-heteroaryl, -alkyl-NH-heterocyclyl and arylalkyl, wherein each of said heterocycloalkyl, heterocycloalkylalkyl, spiroheterocycloalkyl, heterocyclenyl, cycloalkyl, heteroaryl, aryl, alkyl, alkynyl, heterocyclenylalkyl, cycloalkylalkyl, heteroarylalkyl
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl;
  • R 3 is cycloalkyl or heteroaryl, wherein each of said cycloalkyl or heteroaryl can be unsubstituted or substituted with one or more moieties independently selected from the group consisting of X; and R 4 is H.
  • the present invention includes a method of inhibiting mammalian Target Of Rapamycin in a patient, wherein the method comprises administering a therapeutically effective amount of at least one compound of the structural Formula I.
  • the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, wherein the method comprises administering a therapeutically effective amount of at least one compound represented by the structural Formula I.
  • the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, wherein the method comprises administering a therapeutically effective amount of at least one compound represented by the structural Formula I, to a patient in need thereof.
  • the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, and disorders commonly occurring in connection with transplantation, diseases that respond to inhibition of mammalian Target Of Rapamycin, wherein the method comprises administering a therapeutically effective amount of compound represented by the structural Formula I, to a patient in need of such treatment.
  • the present invention includes a method of treatment of a disease, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I.
  • the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, and disorders commonly occurring in connection with transplantation, diseases that respond to inhibition of mammalian Target Of Rapamycin, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I, wherein the disease is a proliferative disease, autoimmune disease, viral disease, fungal disease, neurological/neurodegenerative disorder, arthritis, inflammation, neuronal, alopecia or cardiovascular disease.
  • a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, and disorders commonly occurring in connection with transplantation, diseases that respond to inhibition of mammalian Target Of Rapamycin
  • the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I, wherein the disease is a proliferative disease, autoimmune disease, viral disease, fungal disease, neurological/neurodegenerative disorder, arthritis, inflammation, neuro
  • the present invention includes a method of treatment of a disease, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I, wherein the disease is a proliferative disease.
  • the present invention includes a method of treatment of a proliferative disease, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I, wherein the proliferative disease is selected from the group consisting of: cancer of the bladder, breast, colon, kidney, liver, lung, small cell lung cancer, non- small cell lung cancer, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma, Burkett's lymphoma; acute and chronic myelogenous leukemia, myelodysplastic syndrome, promyelocytic leukemia, mye
  • the present invention includes a method of treatment of a proliferative disease comprising administering a therapeutically effective amount of the compound represented by the structural Formula I, further comprising treatment with radiation therapy.
  • the present invention includes a method of inhibiting a mammalian Target Of Rapamycin, wherein the method comprises administering a therapeutically effective amount of a compound represented by the structural Formula I.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R 1 is heterocycloalkyl, wherein said heterocycloalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl- C(O) 2 H, -alkyl(CO)N(CH 3 )-O-CH 3 , -alkyl(CO)-heteroaryl, -C(O) 2 -alkyl, -alkyl-C(O)-NH 2 , -NH 2 , heteroaryl, -alkyl-CN, -C(O) 2
  • R 1 is heterocyclenyl, wherein said heterocyclenyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl- C(O) 2 H, -alkyl(CO)N(CH 3 )-O-CH 3 , -alkyl(CO)-heteroaryl, -C(O) 2 -alkyl, -alkyl-C(O)-NH 2 , -NH 2 , heteroaryl, -alkyl-CN, -C(O) 2
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CHa) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R 1 is heteroaryl, wherein said heteroaryl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl - C(O) 2 H, -alkyl(CO)N(CH 3 )-O-CH 3 , -alkyl(CO)-heteroaryl, -C(O) 2 -alkyl
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R 1 is cycloalkyl, wherein said cycloalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl-
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of halo, hydroxyl, amino,
  • R 1 is cycloalkenyl, wherein said cycloalkenyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl- C(O) 2 H, -alkyl(CO)N(CH 3 )-O-CH 3 , -alkyl(CO)-heteroaryl, -C(O) 2 -alkyl, -alkyl-C(O)-NH 2 , -NH 2 , heteroaryl, -alkyl-CN, -C(O) 2
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heteroeycloalkyl, and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R 1 is aryl, wherein said aryl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heteroeycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl- C(O) 2 H, -alkyl(CO)N(CH 3 )-O-CH 3 , -alkyl (CO)-heteroaryl, -C(O) 2 -alkyl
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R 1 is alkyl, wherein said alkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl-
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of halo, hydroxyl, amino,
  • R 1 is alkynyl, wherein said alkynyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(0) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl- C(O) 2 H, -alkyl(CO)N(CH 3 )-O-CH 3 , -alkyl(CO)-heteroaryl, -C(O) 2 -alkyl, -alkyl-C(O)-NH 2 , -NH 2 , heteroaryl, -alkyl-CN, -C(O) 2 -
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R 1 is spiroheterocycloalkyl, wherein said spiroheterocycloalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
  • X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O) 2 alkyl, -C(O) 2 H, hydroxyalkyl, -S(O) 2 alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O) 2 -alkyl, -C(O)-heteroaryl, -alkyl- C(O) 2 H, -alkyl(CO)N(CH 3 )-O-CH 3 , -alkyl(CO)-heteroaryl, -C(O) 2 -alkyl, -alkyl-C(O)-NH 2 , -NH 2 , heteroaryl, -alkyl-CN, -C(O) 2
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R 1 is independently selected from the group consisting of:
  • R 2 is quinolinyl, 1 -methyl-pyrazolyl, naphtyl, methoxy-naphtyl, cyano-phenyl, methoxy-phenyl, phenoxy-phenyl, biphenyl, chloro-pyridyl, methoxy-pyridyl, bromo- methyl-pyrazolyl, hydroxyl-methoxy-phenyl, di-methoxy-phenyl, 1 H-indazolyl, fluoro- hydroxy-phenyl, chloro-quinolyl, morpholino-pyridyl-, 4-(isobutyronitrilo)-phenyl, bromo-quinolyl, 4-trifluoromethyl-phenyl-, benzyl -pyrazolyl, fluoro-pyridyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1 -methyl-pyrazolyl, and thienyl;
  • R 1 is independently selected from the group consisting of:
  • R 2 is indazolyl, wherein said indazolyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R 1 is independently selected from the group consisting of:
  • R 2 is phenyl, wherein said phenyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo, alkyl, alkoxy, -CN, hydroxyl, aryl and heteroaryl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R 1 is independently selected from the group consisting of:
  • R 2 is naphthyl, wherein said naphthyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R 1 is independently selected from the group consisting of:
  • R 2 is quinolinyl, wherein said quinolinyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R 1 is independently selected from the group consisting of:
  • R 2 is pyrazolyl, wherein said pyrazolyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R 1 is independently selected from the group consisting of:
  • R 2 is pyridinyl, wherein said pyridinyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo, alkyl, heterocyclyl, heteroaryl, aryl and alkoxy.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is halo
  • R 1 is independently selected from the group consisting of:
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, heteroaryl and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is thienyl; R 1 is independently selected from the group consisting of:
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ⁇ CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, heteroaryl and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is pyrazolyl, wherein said pyrazolyl can be unsubstituted or substituted with alkyl;
  • R 1 is independently selected from the group consisting of:
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is pyridinyl; R 1 is independently selected from the group consisting of:
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyljieterocycloalkyl, heteroaryl and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is methyl; R 1 is independently selected from the group consisting of:
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CHa) 2 CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
  • R is phenyl, wherein said phenyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl;
  • R 1 is independently selected from the group consisting of:
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is acetyl; R 1 is independently selected from the group consisting of:
  • R 2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyljieterocycloalkyl, heteroaryl and arylalkyl.
  • the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
  • R is -CN; R 1 is independently selected from the group consisting of: and
  • R 2 is h ⁇ teroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH 3 ) 2 CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
  • Non-limiting examples of the compounds of the present invention include.
  • the present invention relates to a compound represented by the structural Formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof, in purified form.
  • the present invention relates to a compound represented by the structural Formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof, in isolated form.
  • the present invention includes a composition comprising a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, in combination with at least one pharmaceutically acceptable carrier.
  • the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, said method comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above, or a pharmaceutically acceptable salt, solvate, ester or prodrug of the compound, to a patient in need thereof.
  • the present invention includes a composition comprising a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof, and an anti-cancer agent.
  • the present invention includes a composition comprising a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof, and an anti-neoplastic, anti-tumor, anti-angiogenic, or chemotherapeutic agent.
  • the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, and disorders commonly occurring in connection with transplantation, diseases that respond to inhibition of nnTOR, comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
  • the present invention includes a method of treatment of a proliferative disease, autoimmune disease, viral disease, fungal disease, neurological/neurodegenerative disorder, arthritis, inflammation, neuronal, alopecia or cardiovascular disease comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
  • the present invention includes a method of treatment of a proliferative disease.
  • the present invention includes a method of treatment of a proliferative disease, wherein the proliferative disease is selected from the group consisting of: cancer of the bladder, breast, colon, kidney, liver, lung, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin; small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma, Burkett's lymphoma; acute and chronic myelogenous leukemia, myelodysplastic syndrome, promyelocyte leukemia; fibrosarcoma, rhabdomyosarcoma; astrocytom
  • the present invention includes a method of treatment of hamartoma syndromes, transplant rejection, bowel disorders, inflammatory bowel disease, multiple sclerosis, immunosuppression, immune tolerance, autoimmune diseases, inflammation, bone loss, rheumatoid arthritis, restinosis, cardiac allograft vasculopathy, psoriasis, ocular conditions such as dry eye, hepatic fibrosis, hepatic necrosis, beta-thalassaemia, comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
  • the present invention includes a method of treatment of cancer of the bladder, breast, colon, kidney, liver, lung, small cell lung cancer, non- small cell lung cancer, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma, Burkett's lymphoma; acute and chronic myelogenous leukemia, myelodysplastic syndrome, promyelocytic leukemia; fibrosarcoma, rhabdomyosarcoma; astrocytoma, neuroblastoma, glioma and schwannomas; melanoma
  • the present invention includes a method of treating a disease by inhibiting a mTOR, comprising administering to a patient in need of such treatment an amount of a first compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof; and an amount of at least one second compound, said second compound being an anti-cancer agent; wherein said anti-cancer agent is selected from the group consisting of Adriamycin, Altretamine, Amidox, Aminoglutethimide, Amsacrine, Anastrazole, Antibodies to EGFR, 3-AP, Aphidicolon, Ara-C, Arsenic trioxide, L-Asparaginase, Bevacizumab, Bleomycin, BMS 214662, Bortezomib, Busulfan, Campath, Camptostar, Capecitabine, Carboplatin, Carmustine, Centrosome associated protein E (“CENP-E”) inhibitors, Cetuximab, Cladrib
  • the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, said method comprising administering a therapeutically effective amount of at least one compound of Formula I, or a pharmaceutically acceptable salt, solvate, ester or prodrug of the compound, to a patient in need thereof.
  • the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, diseases related to transplant rejection and diseases that respond to inhibition of mTOR, comprising administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
  • a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, diseases related to transplant rejection and diseases that respond to inhibition of mTOR
  • the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, diseases related to transplant rejection and diseases that respond to inhibition of mTOR, comprising administering a therapeutically effective amount of at least one compound of Formula I or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
  • the present invention includes a compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof exhibiting mTOR inhibition which is at least five-fold the inhibition of CDK2 or CHK-1 by said compound.
  • the present invention includes a compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof exhibiting mTOR inhibition which is at least ten-fold the inhibition of CDK2 or CHK-1 by said compound.
  • the present invention includes a compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof exhibiting mTOR inhibition which is at least fifty-fold the inhibition of CDK2 or CHK-1 by said compound.
  • the compounds of this invention can be used to inhibit the following kinases:
  • Alkyl means an aliphatic hydrocarbon group which may be straight or branched and comprising about 1 to about 20 carbon atoms in the chain. Preferred alkyl groups contain about 1 to about 12 carbon atoms in the chain. More preferred alkyl groups contain about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain.
  • Lower alkyl means a group having about 1 to about 6 carbon atoms in the chain which may be straight or branched.
  • alkyl groups include methyl, ethyl, n-propyl, isopropyl and t-butyl.
  • alkenyl means an aliphatic hydrocarbon group containing at least one carbon- carbon double bond and which may be straight or branched and comprising about 2 to about 15 carbon atoms in the chain. Preferred alkenyl groups have about 2 to about 12 carbon atoms in the chain; and more preferably about 2 to about 6 carbon atoms in the chain. Branched means that one or more, lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkenyl chain.
  • “Lower alkenyl” means about 2 to about 6 carbon atoms in the chain which may be straight or branched.
  • “Alkenyl” may be unsubstituted or optionally substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of halo, alkyl. aryl, cycloalkyl, cyano, alkoxy and -S(alkyl).
  • suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut- 2-enyl, n-pentenyl, octenyl and decenyl.
  • “Alkylene” means a difunctional group obtained by removal of a hydrogen atom from an alkyl group that is defined above. Non-limiting examples of alkylene include methylene, ethylene and propylene.
  • Alkynyl means an aliphatic hydrocarbon group containing at least one carbon- carbon triple bond and which may be straight or branched and comprising about 2 to about 15 carbon atoms in the chain.
  • Preferred alkynyl groups have about 2 to about 12 carbon atoms in the chain; and more preferably about 2 to about 4 carbon atoms in the chain.
  • Branched means that one or more, lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkynyl chain.
  • Lower alkynyl means about 2 to about 6 carbon atoms in the chain which may be straight or branched.
  • alkynyl groups include ethynyl, propynyl, 2-butynyl and 3- methylbutynyl.
  • Alkynyl may be unsubstituted or optionally substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of alkyl, aryl and cycloalkyl.
  • Aryl means an aromatic monocyclic or multicyclic ring system comprising about 6 to about 14 carbon atoms, preferably about 6 to about 10 carbon atoms. The aryl group can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined herein.
  • Suitable aryl groups include phenyl and naphthyl.
  • “Bridged cyclic ring” is a hydrocarbon ring such as cycloalkyl, cyclenyl, or aryl or heteroatom containing ring such as, heterocyclyl, heterocyclenyl, or heteroaryl as described herein, that contains a bridge, which is a valence bond or an atom or an unbranched chain of atoms connecting two different parts of the ring.
  • bridgeheads The two tertiary carbon atoms connected through the bridge are termed "bridgeheads".
  • Heteroaryl means an aromatic monocyclic or multicyclic ring system comprising about 5 to about 14 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the ring atoms is an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. Preferred heteroaryls contain about 5 to about 6 ring atoms.
  • the "heteroaryl” can be optionally substituted by one or more "ring system substituents" which may be the same or different, and are as defined herein.
  • the prefix aza, oxa or thia before the heteroaryl root name means that at least a nitrogen, oxygen or sulfur atom respectively, is present as a ring atom.
  • heteroaryl may also include a heteroaryl as defined above fused to an aryl as defined above.
  • suitable heteroaryls include pyridyl, pyrazinyl, furanyl, thienyl, pyrimidinyl, pyridone (including N-substituted pyridones), isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl, pyrazolyl, triazolyl, 1 ,2,4- thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl, imidazo[1 ,2- a]pyridinyl, imidazo[2,1 -b]thiazolyl
  • Aralkyl or “arylalkyl” means an aryl-alkyl- group in which the aryl and alkyl are as previously described. Preferred aralkyls comprise a lower alkyl group. Non-limiting examples of suitable aralkyl groups include benzyl, 2-phenethyl and naphthalenylmethyl. The bond to the parent moiety is through the alkyl.
  • Alkylaryl means an alkyl-aryl- group in which the alkyl and aryl are as previously described. Preferred alkylaryls comprise a lower alkyl group. Non-limiting example of a suitable alkylaryl group is tolyl. The bond to the parent moiety is through the aryl.
  • Cycloalkyl means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7 ring atoms.
  • the cycloalkyl can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined above.
  • suitable monocyclic cycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.
  • suitable multicyclic cycloalkyls include 1 -decalinyl, norbomyl, adamantyl and the like.
  • Cycloalkylalkyl means a cycloalkyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core.
  • suitable cycloalkylalkyls include cyclohexylmethyl, adamantylmethyl and the like.
  • Cycloalkenyl means a non-aromatic mono or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms which contain at least one carbon-carbon double bond. Preferred cycloaikenyl rings contain about 5 to about 7 ring atoms.
  • the cycloalkenyl can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined above.
  • suitable monocyclic cycioalkenyls include cyclopentenyi, cyclohexenyl, cyclohepta-1 ,3-dienyl, and the like.
  • Non-iimiting example of a suitable multicyclic cycloalkenyl is norbornylenyl.
  • Cycloalkenylalkyl means a cycloalkenyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core.
  • suitable cycloalkenyialkyls include cyclopentenylmethyl, cyclohexenylmethyl and the like.
  • Halogen means fluorine, chlorine, bromine, or iodine. Preferred are fluorine, chlorine and bromine.
  • Ring system substituent means a substituent attached to an aromatic or non- aromatic ring system which, for example, replaces an available hydrogen on the ring system.
  • Ring system substituents may be the same or different, each being independently selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryi, araikyl, alkylaryl, heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkyl heteroaryl, hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo, nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyi, aralkoxycarbonyl, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaral
  • Ring system substituent may also mean a single moiety which simultaneously replaces two available hydrogen on two adjacent carbon atoms (one H on each carbon) on a ring system.
  • Examples of such moiety are methylene dioxy, ethyl enedioxy, -C(CH 3 J 2 - and the like which form moieties such as, for example:
  • Heteroarylalky means a heteroary! moiety as defined above linked via an alkyl moiety (defined above) to a parent core.
  • suitable heteroaryis include 2-pyridinylmethyl, quinolinylmethy! and the like.
  • Heterocyciyl means a non-aromatic saturated monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the atoms in the ring system is an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and/or sulfur atoms present in the ring system.
  • Preferred heterocyclyls contain about 5 to about 6 ring atoms.
  • the prefix aza, oxa or thia before the heterocyciyl root name means that at least a nitrogen, oxygen or sulfur atom respectively is present as a ring atom.
  • Any -NH in a heterocyciyl ring may exist protected such as, for example, as an -N(Boc), -N(CBz), -N(Tos) group and the like; such protections are also considered part of this invention.
  • the heterocyciyl can be optionally substituted by one or more "ring system substituents" which may be the same or different, and are as defined herein.
  • the nitrogen or sulfur atom of the heterocyciyl can be optionally oxidized to the corresponding N-oxide, S-oxide or S 1 S- dioxide.
  • suitable monocyclic heterocyciyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyi, thiazolidinyl, 1 ,4- dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, lactam, lactone, and the like.
  • Heterocyciyl may also mean a single moiety (e.g., carbonyl) which simultaneously replaces two available hydrogen on the same carbon atom on a ring system.
  • a single moiety e.g., carbonyl
  • pyrrolidone e.g., pyrrolidone
  • Heterocyclylalkyl means a heterocyclyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core.
  • suitable heterocyclylaikyls include piperidinylmethyl, piperazinylmethyl and the like.
  • Heterocyclenyl means a non-aromatic monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the atoms in the ring system is an element other than carbon, for example nitrogen, oxygen or sulfur atom, alone or in combination, and which contains at least one carbon-carbon double bond or carbon-nitrogen double bond. There are no adjacent oxygen and/or sulfur atoms present in the ring system.
  • Preferred heterocycienyl rings contain about 5 to about 6 ring atoms.
  • the prefix aza, oxa or thia before the heterocyclenyl root name means that at least a nitrogen, oxygen or sulfur atom respectively is present as a ring atom.
  • the heterocycienyl can be optionally substituted by one or more ring system substituents, wherein "ring system substituent" is as defined above.
  • the nitrogen or sulfur atom of the heterocyclenyl can be optionally oxidized to the corresponding N-oxide, S-oxide or S,S-dioxide.
  • heterocyclenyl groups include 1 ,2,3,4- tetrahydropyridinyl, 1 ,2-dihydropyridinyl, 1 ,4-dihydropyridinyl, 1 ,2,3,6- tetrahydropyridinyl, 1 ,4,5,6-tetrahydropyrimidinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2- imidazolinyl, 2-pyrazoiinyl, dihydroimidazolyl, dihydrooxazolyi, dihydrooxadiazolyl, dihydrothiazolyl, 3,4-dihydro ⁇ 2H-pyranyS, dihydrofuranyl, fluorodihydrofuranyi, 7- oxabicyclo[2.2.1]heptenyl, dihydrothiophenyi, dihydrothiopyranyl, and the like.
  • Heterocyclenyl may also mean a single moiety
  • Heterocyclenylalkyl means a heterocyclenyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core. It should be noted that in hetero-atom containing ring systems of this invention, there are no hydroxyl groups on carbon atoms adjacent to a N, O or S, as well as there are no N or S groups on carbon adjacent to another heteroatom. Thus, for example, in the ring:
  • Alkynylalkyl means an alkynyi-alkyl- group in which the alkynyl and alkyl are as previously described. Preferred alkynylalkyls contain a lower alkynyi and a lower alkyl group. The bond to the parent moiety is through the alkyf. Non-limiting examples of suitable alkynylalkyl groups include propargyimethyl. "Heteroaralkyi” means a heteroaryl-alkyl- group in which the heteroaryl and alkyl are as previously described. Preferred heteroaralkyls contain a lower alky! group. Non-limiting examples of suitable araikyl groups include pyridyl methyl, and quinolin-3- ylmethyl. The bond to the parent moiety is through the alkyi.
  • Spiro ring systems have two or more rings linked by one common atom.
  • Preferred spiro ring systems include spiroheteroaryl, spiroheterocyclenyl, spiroheterocyclyl, spirocycloaikyl, spirocyclenyl, and spiroaryl.
  • the spiro ring systems can be optionally substituted by one or more ring system substituents, wherein "ring system substituent" is as defined above.
  • systems include spiro[4.5jdecane, 8-azaspiro[4.5]dec-2-ene, and ' spiro[4.4]nona-2 t 7-diene.
  • ⁇ ydroxyalkyl means a HO-aikyl- group in which alkyl is as previously defined.
  • Preferred hydroxyalkyls contain lower alkyl.
  • suitable hydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl.
  • acyl means an H-C(O)-, alkyl-C(O)- or cycloalkyl-C(O)-, group in which the various groups are as previously described.
  • the bond to the parent moiety is through the carbonyl.
  • Preferred acyls contain a lower alkyl.
  • suitable acyl groups include formyl, acetyl and propanoyl.
  • Aroyl means an aryl-C(O)- group in which the aryl group is as previously described.
  • the bond to the parent moiety is through the carbonyi.
  • suitable groups include benzoyl and 1- naphthoyl.
  • Alkoxy means an alkyl-O- group in which the alky! group is as previously described.
  • suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy.
  • the bond to the parent moiety is through the ether oxygen.
  • Aryloxy means an aryl-O- group in which the aryl group is as previously described. Non- ⁇ miting examples of suitable aryloxy groups include phenoxy and naphthoxy. The bond to the parent moiety is through the ether oxygen.
  • Aralkyloxy means an aralkyl-O- group in which the aralkyi group is as previously described. Non-limiting examples of suitable aralkyloxy groups include benzyloxy and 1- or 2-naphthalenemethoxy. The bond to the parent moiety is through the ether oxygen.
  • Alkylthio means an alkyl-S- group in which the aikyl group is as previously described.
  • suitable alkylthio groups include methylthio and ethylthio.
  • the bond to the parent moiety is through the sulfur.
  • Arylthio means an aryl-S- group in which the aryl group is as previously described.
  • suitable arylthio groups include phenyithio and naphthylthio.
  • the bond to the parent moiety is through the sulfur.
  • Aralkylthio means an aralkyl-S- group in which the aralkyl group is as previously described.
  • Non-limiting example of a suitable aralkylthio group is benzylthio. The bond to the parent moiety is through the sulfur.
  • Alkoxycarbonyl means an alkyi-O-CO- group.
  • suitable alkoxycarbonyl groups include methoxycarbonyl and ethoxycarbonyl.
  • the bond to the parent moiety is through the carbonyl.
  • Aryloxycarbonyl means an aryl-O-C(O)- group.
  • suitable aryloxycarbonyi groups include phenoxycarbonyl and naphthoxycarbonyi.
  • the bond to the parent moiety is through the carbonyl.
  • Aralkoxycarbonyl means an aralkyl-O-C(O)- group.
  • Non-limiting example of a suitable aralkoxycarbonyl group is benzyloxycarbonyl. The bond to the parent moiety is through the carbonyl.
  • Alkylsulfonyi means an alkyi-S(O 2 )- group. Preferred groups are those in which the alkyl group is lower alkyl. The bond to the parent moiety is through the sulfonyl.
  • Arylsulfonyl means an aryl-S(O 2 )- group. The bond to the parent moiety is through the sulfonyl.
  • substituted means that one or more hydrogen on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency under the existing circumstances is not exceeded, and that the substitution results in a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.
  • stable compound' or “stable structure” is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
  • purified refers to the physical state of said compound after being isolated from a synthetic process (e.g. from a reaction mixture), or natural source or combination thereof.
  • purified refers to the physical state of said compound after being obtained from a purification process or processes described herein or wefl known to the skilled artisan (e.g., chromatography, recrystallization and the like) , in sufficient purity to be characterizable by standard analytical techniques described herein or well known to the skilled artisan. It should aiso be noted that any carbon as well as heteroatom with unsatisfied valences in the text, schemes, examples and Tables herein is assumed to have the sufficient number of hydrogen atom(s) to satisfy the valences.
  • protecting groups When a functional group in a compound is termed "protected", this means that the group is in modified form to preclude undesired side reactions at the protected site when the compound is subjected to a reaction. Suitable protecting groups will be recognized by those with ordinary skill in the art as well as by reference to standard textbooks such as, for example, T. W. Greene et a/, Protective Groups in organic Synthesis (1991), Wiley, New York.
  • variable e.g., aryl, heterocycle, R 2 , etc.
  • its definition on each occurrence is independent of its definition at every other occurrence.
  • composition is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
  • Prodrugs and solvates of the compounds of the invention are also contemplated herein.
  • a discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press.
  • the term "prodrug” means a compound (e.g., a drug precursor) that is transformed in vivo Xo yieid a compound of the present invention or a pharmaceutically acceptable salt, hydrate or solvate of the compound. The transformation may occur by various mechanisms (e.g., by metabolic or chemical processes), such as, for example, through hydrolysis in blood.
  • a prodrug can comprise an ester formed by the replacement of the hydrogen atom of the acid group with a group such as, for example, (C 1 -C ⁇ jalkyl, (C 2 - Ci 2 )alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1- methyl-1 -(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms, 1 -(N-(alk)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1 -(N-
  • a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a group such as, for example, (C 1 -C 6 )alkanoyloxymethyl, 1 -((Ci- C 6 )alkanoyloxy)ethyl, 1 -methyl-1 -((C 1 -C 6 )alkanoyloxy)ethyl, (C 1 - C 6 )alkoxycarbonyloxymethyl, N-(C 1 -C 6 )alkoxycarbonylaminomethyl, succinoyl, (Cr C 6 )alkanoy!, ⁇ -amino(C 1 -C 4 )alkanyl, aryiacyl and ⁇ -aminoacyl, or ⁇ -aminoacyl- ⁇ - aminoacyl, where each ⁇ -aminoacyl group is independently selected from the naturally occurring L-amino acids, P(O)
  • a prodrug can be formed by the replacement of a hydrogen atom in the amine group with a group such as, for example, R-carbonyi, RO-carbonyl, NRR'-carbonyl where R and R' are each independently (Ci ⁇ Cio)alkyt, (C 3 -C 7 ) cycloalkyl, benzyl, or R-carbonyl is a natural ⁇ -aminoacyl or natural ⁇ -aminoacyl, -C(OH)C(O)OY 1 wherein Y 1 is H, (C 1 -Ce)alkyl or benzyl, -C(OY 2 JY 3 wherein Y 2 is (C 1 -C 4 ) alky] and Y 3 is (C 1 -C 6 )alkyl, carboxy (C 1 -C 6 )alky!, amino(C 1 -C 4 )alkyl or mono-N —
  • R-carbonyi RO-carbonyl
  • One or more compounds of the invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable sêts such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms.
  • “Solvate” means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. "Solvate” encompasses both solution-phase and isolatable solvates. Non-limiting examples of suitable solvates include ethanoiates, methanolates, and the like.
  • “Hydrate” is a solvate wherein the solvent molecule is H 2 O.
  • One or more compounds of the invention may optionally be converted to a solvate.
  • Preparation of solvates is generally known.
  • M. Caira et al, J. Pharmaceutical Sci., 93(3), 601-611 (2004) describes the preparation of the solvates of the antifungal fluconazole in ethyl acetate as well as from water.
  • Similar preparations of solvates, hemisolvate, hydrates and the like are described by E. C. van Tonder et al, AAPS PharmSciTech., 5£1), article 12 (2004); and A. L. Bingham et al, Chem. Comm ⁇ n., 603-604 (2001).
  • a typical, non-limiting, process involves dissolving the inventive compound in desired amounts of the desired solvent (organic or water or mixtures thereof) at a higher than ambient temperature, and cooling the solution at a rate sufficient to form crystals which are then isolated by standard methods.
  • Analytical techniques such as, for example I. R. spectroscopy, show the presence of the solvent (or water) in the crystals as a solvate (or hydrate).
  • Effective amount or “therapeutically effective amount” is meant to describe an amount of compound or a composition of the present invention effective in inhibiting the above-noted diseases and thus producing the desired therapeutic, ameliorative, inhibitory or preventative effect.
  • the compounds of the present invention can form salts which are also within the scope of this invention.
  • Reference to a compound of the present invention herein is understood to include reference to salts thereof, unless otherwise indicated.
  • the term “saltfs)" denotes acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases.
  • salts when a compound of the present invention contains both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions ("inner salts") may be formed and are included within the term "salt(s)" as used herein.
  • Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful.
  • Salts of the compounds of the The present invention may be formed, for example, by reacting a compound of the present invention with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.
  • Exemplary acid addition salts include acetates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromides, hydroiodides, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartarates, thiocyanates, toluenesulfonates (also known as tosylates,) and the like.
  • Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamines, t-butyl amines, and salts with amino acids such as arginine, lysine and the like.
  • Basic nitrogen-containing groups may be quartemized with agents such as lower alkyl hatides (e.g. methyl, ethyl, and butyl chlorides, bromides and iodides), dialky! sulfates (e.g.
  • dimethyl, diethyl, and dibutyl sulfates dimethyl, diethyl, and dibutyl sulfates
  • long chain halides e.g. decyl, iauryl, and stearyl chlorides, bromides and iodides
  • aralkyl haiides e.g. benzyl and phenethyl bromides
  • esters of the present compounds include the following groups: (1 ) carboxylic acid esters obtained by esterification of the hydroxy groups, in which the non-carbonyl moiety of the carboxyiic acid portion of the ester grouping is selected from straight or branched chain alkyl (for example, acetyl, n- propyl, t-butyl, or n-butyl), alkoxyalkyl (for example, methoxymethyl), araikyl (for example, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (for example, phenyl optionally substituted with, for example, halogen, d ⁇ alkyl, or Ci.
  • alkyl for example, acetyl, n- propyl, t-butyl, or n-butyl
  • alkoxyalkyl for example, methoxymethyl
  • araikyl for example, benzyl
  • aryloxyalkyl for example,
  • sulfonate esters such as alkyl- or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters (for example, L-valyl or L-isoieucyl); (4) phosphonate esters and (5) mono-, di- or triphosphate esters.
  • the phosphate esters may be further esterified by, for example, a C 1-2 o alcohol or reactive derivative thereof, or by a 2,3-di (C 6 - 24 )acyl glycerol.
  • the compounds of the present invention may contain asymmetric or chiral centers, and, therefore, exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the present invention as well as mixtures thereof, including racemic mixtures, form part of the present invention.
  • the present invention embraces all geometric and positional isomers. For example, if a compound of the present invention incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the invention.
  • Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods well known to those skilled in the art, such as, for example, by chromatography and/or fractional crystallization.
  • Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydroiyzing) the individual diastereomers to the corresponding pure enantiomers.
  • an appropriate optically active compound e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride
  • converting e.g., hydroiyzing
  • some of the compounds of the present invention may be atropisomers (e.g., substituted biaryls) and are considered as part of this invention.
  • the compounds of the present invention may exist in different tautomeric forms, and all such forms are embraced within the scope of the invention. Also, for example, all keto-enol and imine-enamine forms of the compounds are included in the invention.
  • All stereoisomers for example, geometric isomers, optical isomers and the like
  • of the present compounds ⁇ including those of the salts, solvates, esters and prodrugs of the compounds as well as the salts, solvates and esters of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated within the scope of this invention, as are positional isomers (such as, for example, 4-pyridyl and 3-pyridyl).
  • a compound of the present invention incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the invention. Also, for example, all keto-enol and imine-enamine forms of the compounds are included in the invention.
  • Individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers.
  • the chiral centers of the present invention can have the S or R configuration as defined by the IUPAC 1974 Recommendations.
  • salt is intended to equally apply to the salt, solvate, ester and prodrug of enantiomers, stereoisomers, rotamers, tautomers, positional isomers, racemates or prodrugs of the inventive compounds.
  • the present invention also embraces isotopically-labelled compounds of the present invention which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S 1 18 F, 36 CI and 123 I, respectively.
  • Certain isotopically-labelled compounds of Formula (!) are useful in compound and/or substrate tissue distribution assays. Tritiated ⁇ i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred for their ease of preparation and detectabiiity. Certain isotopically-labelled compounds of Formula (I) can be useful for medical imaging purposes.
  • those labeled with positron-emitting isotopes like 11 C or 18 F can be useful for application in Positron Emission Tomography (PET) and those labeled with gamma ray emitting isotopes like 123 I can be useful for application in Single photon emission computed tomography (SPECT).
  • PET Positron Emission Tomography
  • SPECT Single photon emission computed tomography
  • substitution with heavier isotopes such as deuterium (i.e., 2 H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances.
  • isotopically labeled compounds of Formula (I) in particular those containing isotopes with longer half lives (T1/2 >1 day), can generally be prepared by following procedures analogous to those disclosed in the Schemes and/or in the Examples herein below, by substituting an appropriate isotopically labeled reagent for a non-isotopically labeled reagent.
  • polymorphic forms of the compounds of the present invention, and of the salts, solvates, esters and prodrugs of the compounds of the present invention, are intended to be included in the present invention.
  • the compounds according to the invention have pharmacological properties; in particular, the compounds of the present invention can be inhibitors, regulators or modulators of mTOR protein kinases.
  • the compounds of the present invention can be inhibitors of protein kinases such as, for example, the inhibitors of the mTOR.
  • Preferred compounds can exhibit IC 50 values of less than about 5 ⁇ m, preferably about 0.001 to about 1.0 ⁇ m, and more preferably about 0.001 to about 0.1 ⁇ m.
  • the assay methods are described in the Examples set forth below.
  • the compounds of the present invention can be useful in the therapy of proliferative diseases such as cancer, autoimmune diseases, viral diseases, fungal diseases, neurological/neurodegenerative disorders, arthritis, inflammation, antiproliferative (e.g., ocular retinopathy), neuronal, alopecia and cardiovascular disease.
  • proliferative diseases such as cancer, autoimmune diseases, viral diseases, fungal diseases, neurological/neurodegenerative disorders, arthritis, inflammation, antiproliferative (e.g., ocular retinopathy), neuronal, alopecia and cardiovascular disease.
  • proliferative diseases such as cancer, autoimmune diseases, viral diseases, fungal diseases, neurological/neurodegenerative disorders, arthritis, inflammation, antiproliferative (e.g., ocular retinopathy), neuronal, alopecia and cardiovascular disease.
  • the compounds of the present invention can be useful in the treatment of a variety of cancers, including (but not limited to) the following: tumor of the bladder, breast (including BRCA-mutated breast cancer, colorectal, colon, kidney, liver, lung, small cell lung cancer, non-smail cell lung cancer, head and neck, esophagus, bladder, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, including squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma and Burkett's lymphoma; chronic lymphocytic leukemia ("CLL”), acute and chronic myelogenous leukemia, myelodysplastic syndrome and prom
  • CLL chronic lympho
  • inhibitors could act as reversible cytostatic agents which may be useful in the treatment of any disease process which features abnormal cellular proliferation, e.g., benign prostate hyperplasia, familial adenomatosis polyposis, neuro-fibromatosis, atherosclerosis, pulmonary fibrosis, arthritis, psoriasis, glomerulonephritis, restenosis following angioplasty or vascular surgery, hypertrophic scar formation, inflammatory bowel disease, transplantation rejection, endotoxic shock, and fungal infections.
  • Compounds of the present invention may induce or inhibit apoptosis.
  • apoptotic response is aberrant in a variety of human diseases.
  • Compounds of the present invention, as modulators of apoptosis will be useful in the treatment of cancer (including but not limited to those types mentioned hereinabove), virai infections (including but not limited to herpevirus, poxvirus, Epstein- Barr virus, Sindbis virus and adenovirus), prevention of AIDS development in HIV-infected individuals, autoimmune diseases (including but not limited to systemic lupus, erythematosus, autoimmune mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes mellitus), neurodegenerative disorders (including but not limited to Alzheimer's disease, AIDS-reiated dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy and cerebellar degeneration), myelodysplasia syndromes, aplastic anemia, ische
  • Compounds of the present invention can modulate the level of cellular RNA and DNA synthesis. These agents would therefore be useful in the treatment of viral infections (including but not limited to HIV, human papilloma virus, herpesvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus). Compounds of the present invention may also be useful in the chemoprevention of cancer. Chemoprevention is defined as inhibiting the development of invasive cancer by either blocking the initiating mutagenic event or by blocking the progression of pre-malignant cells that have already suffered an insult or inhibiting tumor relapse.
  • Compounds of the present invention may also be useful in inhibiting tumor angiogenesis and metastasis.
  • Another aspect of this invention is a method of treating a mammal ⁇ e.g., human) having a disease or condition associated with mTOR kinases by administering a therapeutically effective amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound to the mammal.
  • a preferred dosage is about 0.001 to 1000 mg/kg of body weight/day of the compound of the present invention.
  • An especially preferred dosage is about 0.01 to 25 mg/kg of body weight/day of a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound.
  • the compounds of this invention may also be useful in combination (administered together or sequentially) with one or more of anti-cancer treatments such as radiation therapy, and/or one or more anti-cancer agents different from the compound of the present invention.
  • the compounds of the present invention can be present in the same dosage unit as the anti-cancer agent or in separate dosage units.
  • Another aspect of the present invention is a method of treating one or more diseases associated with a mTOR protein kinase, comprising administering to a mammal in need of such treatment: an amount of a first compound, which is a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof; and an amount of at least one second compound, the second compound being an anti-cancer agent different from the compound of the present invention, wherein the amounts of the first compound and the second compound result in a therapeutic effect.
  • suitable anti-cancer agent is selected from the group consisting of a Cytostatic agent, Cisplatin, Deforolimus (described in PCT publication No.
  • Triethylenethiophosphoramine Busulfan, Carmustine, Lomustine, Streptozocin, dacarbazine, Floxuridine, Cytarabine, 6-Mercaptopurine, 6-Thioguanine, Fludarabine phosphate, Oxaliplatin, Leucovirin, ELOXATINTM, Vinblastine, Vincristine, Vindesine, Bleomycin, Dactinomycin, Daunorubicin, Doxorubicin, Epirubicin, Idarubicin, Mithramycin, Deoxycoformycin, Mitomycin-C, L-Asparaginase, Teniposide 17oc- Ethinylestradiol, Diethylstilbestroi, Testosterone, Prednisone, Fluoxymesterone, Dromostanolone propionate, Testolactone, Megestrolacetate, Methylprednisoione, Methyltestosterone, Prednisolone, Triamcino
  • such combination products employ the compounds of this invention within the dosage range described herein and the other pharmaceutically active agent or treatment within its dosage range.
  • the CDC2 inhibitor olomucine has been found to act synergistically with known cytotoxic agents in inducing apoptosis (J. CeII ScL, (1995) 108, 2897.
  • Compounds of the present invention may also be administered sequentially with known anticancer or cytotoxic agents when a combination formulation is inappropriate.
  • the invention is not limited in the sequence of administration; compounds of the present invention may be administered either prior to or after administration of the known anticancer or cytotoxic agent.
  • this invention includes combinations comprising an amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof, and an amount of one or more anticancer treatments and anti-cancer agents listed above wherein the amounts of the compounds/ treatments result in desired therapeutic effect.
  • Another aspect of the present invention is a method of inhibiting one or more mTOR protein kinases in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of at least one compound of the present invention or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
  • Another aspect of the present invention is a method of treating, or slowing the progression of, a disease associated with one or more mTOR protein kinases in a patient in need thereof, comprising administering a therapeutically effective amount of at least one compound of the present invention or a pharmaceuticaily acceptable salt, solvate, ester or prodrug thereof.
  • Yet another aspect of the present invention is a method of treating one or more diseases associated with mTOR protein kinases, comprising administering to a mammai in need of such treatment an amount of a first compound, which is a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof; and an amount of at least one second compound, the second compound being an anti-cancer agent, wherein the amounts of the first compound and the second compound result in a therapeutic effect.
  • a first compound which is a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof
  • an amount of at least one second compound the second compound being an anti-cancer agent
  • Another aspect of the present invention is a method of treating, or slowing the progression of, a disease associated with one or more mTOR protein kinases in a patient in need thereof, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising in combination at least one pharmaceutically acceptable carrier and at least one compound according to The present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
  • the mTOR protein kinases to be inhibited can exist in two complexes, mTORC1 and mTORC2.
  • ERK inhibitors include but are not limited to the following compounds: a) Formula H
  • Each Q 1 represents a ring independentiy selected from the group consisting of: cycloalkyl, substituted cycloalkyi, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted ary!, heteroaryl, and substituted heteroaryl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: halo and the R 10 moieties; provided that when Q 1 is ary!, heteroaryl, substituted aryl or substituted heteroaryl then the carbon atoms at the ring junction are not substituted;
  • Q 2 represents a ring selected from the group consisting of: cycloalkyl, substituted cycloaikyl, heterocycloaikyl, and substituted heterocycloalkyl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R 10 moieties;
  • Z 1 represents -(C(R 24 ) 2 ) W - wherein each R 24 is independently selected from the group consisting of: H, alkyl and F, and wherein w is 1 , 2 or 3;
  • Z 2 is selected from the group consisting of: -N(R 44 )-, -O- and -C(R 46 J 2 -; m is 1 to 6; n is 1 to 6; p is 0 to 6; t is O 1 1 , or 2;
  • R 1 is selected from the group consisting of: (I) -CN,
  • R 2 is selected from the group consisting of: (D H 1 )
  • each alkyi is independently selected
  • each alkyl is independently selected
  • each R 48 is independently selected from the group consisting of: H and alkyl
  • each R 3 , R 4 , R 5 , R 6 and R 7 is independently selected from the group consisting of: (I) H 1
  • R 11 is selected from the group consisting of: F, -OH, -CN, -OR 10 , -NHNR 1 R 10 , -SR 10 and heteroaryl;
  • R 12 is selected from the group consisting of: alkyl, aryl, heteroaryl, cycloalkyi, eye! oa Iky I alkyl, heterocycloalkyl and heterocycloalkylaikyi;
  • R 14 is selected from the group consisting of: alkyt, aryl, heteroaryl, cycloalkyi, cycloaikylalkyl-, heterocycloalkyl, alkylheterocycloalkyl, heterocycioalkylalkyl-, aikylheteroaryi- and aikyiaryl-;
  • R 15 is selected from the group consisting of: H, -OH, alkyl, aryl, heteroaryl, cycloalkyl, cycloaikylalkyl-, heterocycloalkyl and heterocycloalkylaikyi-, alkylheteroaryl- and alkylary!-;
  • R 20 represents alkyl
  • R 23 is selected from the group consisting of: H, alkyl, aryl, cycloaikyl, and cycloalkylalkyi-; each R 26 is independently selected from the group consisting of: H and alkyl;
  • R 28 is alkyi; each R 30 is independently selected from the group consisting of: H, alkyl, and F; each R 32 is independently selected from the group consisting of: H and alkyl, and wherein each R 32 is generally H; each R 35 is independently selected from the group consisting of: H and Ci to C 6 aikyl;
  • R 36 is selected from the group consisting of: H, alkyl, and -O-alkyl; each R 38 is independently selected from the group consisting of: H, alkyl, aryi, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalky!
  • R 38 substituted alkyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH 2 , -NO 2 , -CN, -OR 26 , halo, -C(O)-NH-R 28 , -C(O)OR 28 , and -C(O)R 28 , and said R 3 ⁇ substituted aryl, substituted aryialkyl, substituted heteroaryl, substituted hetero
  • R 42 is selected from the group consisting of: alkyl, aryl, heteroaryl, and cycloalkyl
  • R 44 is selected from the group consisting of: H, alkyl, cycloalkyl, and cycloaikylalkyl
  • Each R 46 is independently selected from the group consisting of: H, alkyi, cycloalkyl, and cycloalkylalkyl; and provided that:
  • Q 1 is a substituted ring wherein at least one substitutent is halo; and/or (2) R 2 is substituted alkyl wherein said substituent is -OCH 3 , or R 2 is selected from the group consisting of: -CH 2 OH and -CH 2 OCH 3 ; and/or
  • R 3 , R 4 , R 5 , R 6 , and R 7 is:
  • R 5A is alkyl
  • R 10 is a substituted aryl, and at least one substitutent is selected from the group consisting of: (a) -S(O) t R 38 wherein R 38 is isopropyl,
  • R 20 is isopropyl
  • Each Q 1 represents a ring independently selected from the group consisting of: cycioalkyl, substituted cycioaikyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, and substituted heteroaryl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: haio and the R 10 moieties; provided that when Q 1 is ary!, heteroaryl, substituted aryl or substituted heteroaryl then the carbon atoms at the ring junction are not substituted;
  • Q 2 represents a ring selected from the group consisting of: cycloalkyl, substituted cycloalkyi, heterocycloalkyl, and substituted heterocycloalkyl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R 10 moieties;
  • Z 1 represents - ⁇ C(R 24 ) 2 ) W - wherein each R 24 is independently selected from the group consisting of: H, alkyl and F, and wherein w is 1 , 2 or 3;
  • Z 2 is selected from the group consisting of: -N(R 44 )-, -O- and -C(R 46 ) 2 -;
  • m is 1 to 6;
  • n is 1 to 6;
  • p is 0 to 6;
  • t is 0, 1 , or 2;
  • R 1 is selected from the group consisting of:
  • each R 10 is independently selected, (18) wherein each R 10 is independently selected, (19)
  • each R 10 is independently selected, (22) -C(O)-NR 32 -C(R 18 ) 3 ,
  • R 2 is selected from the group consisting of: (I) H 1
  • substituted alkyl wherein said substituted alkyl is substituted with 1 to 3 substitutents selected from the group consisting of: (a) -OH, (b) -O-alkyl, (c) -O-alkyl substituted with 1 to 3 F atoms, and (d) -N(R 40 ) 2 wherein each R 40 is independently selected from the group consisting of: (i) H, (ii) C 1 -C 3 alkyl, (iii) -CF 3 , and (e) halo, (6) alkynyl,
  • each alkyl is independently selected
  • each R 48 is independently selected from the group consisting of: H and aikyl
  • each R 3 , R 4 , R 5 , R 6 and R 7 is independently selected from the group consisting of:
  • R SA is selected from the group consisting of: halo, -OH, alkyl, -O-alkyl;
  • R 8 is selected from the group consisting of: H, -OH, -N(R 10 ) 2) -NR 10 C(O)R 12 ; each R 9 is independently selected from the group consisting of :halogen, -CN,
  • each R 10 is independently selected from the group consisting of: H, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, heterocycioalkyl, heterocycloalkylalkyl, alkylheteroaryl-, alkylaryl-, substituted alkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloaikyl, substituted cycloalkylalkyl, substituted heterocycioalkyl, substituted heterocycloaikylalkyi, substituted alkylheteroaryl-, substituted alkyiaryl-, heterocycloaikenyl
  • R 10 substituted alkyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH 2 , -NHR 20 , -NO 2 , -CN, -OR 26 , halo, -C(O)-NH-R 26 , -C(O)OR 26 , and -C(O)R 26 , and said R 10 substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroaryialkyl, substituted cycloalkyl, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycloaikylalkyi, substituted alkylheteroaryl- and substituted alkyiaryl- are substituted with 1 to 3 substituents independently selected from the group consisting of: (1 ) -NH 2 , (2) -NO 2 , (3) -CN, (4) -OH, (5)
  • R 11 is selected from the group consisting of: F, -OH, -CN, -OR 10 , -NHNR 1 R 10 , -SR 10 and heteroaryl;
  • R 12 is selected from the group consisting of: alkyl, aryl, heteroaryl, cycioalkyl, cycloalkylalkyl, heterocycloalkyl and heterocycloaikylalkyi;
  • R 14 is selected from the group consisting of: aikyl, aryl, heteroaryl, cycloalkyl, cycloalkylalkyl-, heterocycloalkyl, alkylheterocycloalkyl, heterocycloaikylalkyi-, afkylheteroaryt- and alkyiaryl-;
  • R 15 is selected from the group consisting of: H, -OH, alkyi, aryl, heteroaryl, cycloalkyl, cycloalkylalkyl-, heterocycloaikyl and heterocycioalkylalkyl-, alkylheteroaryi- and aikylaryl-;
  • R 20 represents alkyl
  • R 23 is selected from the group consisting of: H, alkyl, aryl, cycloalkyl, and cycloalkylalkyl-
  • each R 26 is independently selected from the group consisting of: H and alkyi;
  • R 28 is alkyl; each R 30 is independently selected from the group consisting of: H 1 alkyl, and F; each R 32 is independently selected from the group consisting of: H and alkyl; each R 35 is independently selected from the group consisting of: H and Ci to C 6 alkyl; each R 38 is independently selected from the group consisting of: H, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyi, heterocycloalkylalky], alkylheteroaryl-, alkylaryl-, substituted alkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloalkyl, substituted cycloalkylalkyl, substituted heterocycloaikyl, substituted heterocycioalkylalkyl, substituted alkylheteroaryl-
  • R 42 is selected from the group consisting of: alkyi, aryl (e.g., phenyl), heteroaryl, and cycloalkyi;
  • R 44 is selected from the group consisting of: H, alkyl, cycloalkyi, and cycloalkylaikyl;
  • Each R 46 is independently selected from the group consisting of: H, alkyl, cycloalkyi, and cycloalkylaikyl;
  • Y 1 , Y 2 , and Y 3 are each independently selected from the group consisting of:
  • Each Q 1 represents a ring independently selected from the group consisting of: cycloalkyl, substituted cycloalkyl, heterocycioalkyl, substituted heterocycloalkyi, aryl, substituted aryl, heteroaryl, and substituted heteroaryl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R 10 moieties; provided that when Q 1 is aryl, heteroaryl, substituted aryl or substituted heteroaryl then the carbon atoms at the ring junction are not substituted;
  • Q 2 represents a ring selected from the group consisting of: cycloalkyi, substituted cycloalkyi, heterocycJoalkyi, and substituted heterocycioalkyl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R 10 moieties;
  • Z 1 represents -(C(R 24 ) 2 )w- wherein each R 24 is independently selected from the group consisting of: H, alkyl and F, and wherein w is 1 , 2 or 3, and generally w is 1 or 2, and usually w is 1 , and wherein in one example each R 24 is H, and in another example w is 1 , and in another example each R 24 is H and w is 1 , preferably w is 1 and each R 24 is H;
  • Z 2 is selected from the group consisting of: -N(R 44 )-, -O- and -C(R 46 J 2 -; m is 1 to 6; n is 1 to 6; p is 0 to 6; t is 0, 1 , or 2;
  • R 1 is selected from the group consisting of: (I) -CN,
  • each R 10 is independently selected, and preferably each R 10 is independently selected from the group consisting of: (a) H, (b) alkyl, (c) heteroaryl, (d) aryl, and (e) cycloalkyl, wherein for example, each R 10 is selected from the group consisting of: H, methyl, butyl, i-propyl, pyridyl, phenyl and cyclopropyl, wherein, for example, said -C(O)N(R 10 ) 2 moiety is selected from the group consisting of: -C(O)NH 2 , -C(O)NH(CH 3 ), -C(O)NH(CH)(CH 3 ) 2J -C(O)NH(C 4 H 9 ), -C(O)NH(C 6 H 5 ), -C(O)NH(C 3 H 5 ), and -C(O)NH(C 5 H 4 N);
  • R 2 is selected from the group consisting of: (1) H,
  • substituted alkyl wherein said substituted alkyl is substituted with 1 to 3 substitutents selected from the group consisting of: (a) -OH, (b) -O-alkyl (e.g., -0-(Cr C 3 alkyl), (c) -O-alkyi (e.g., -O-(C r C 3 alkyl)) substituted with 1 to 3 F atoms, and (d) - N(R 40 )2 wherein each R 40 is independently selected from the group consisting of: (i) H, (ii) CrC 3 alkyl and (iii) -CF 3 ,
  • each R 3 , R 4 , R 5 , R 6 and R 7 is independently selected from the group consisting of:
  • R 10 is selected from the group consisting of: alkyl,
  • arylheteroaryl- (16) substituted arylheteroaryl-, (17) heteroarylaryi-, such as, for example, pyrimidinylphenyl-, pyrazinylphenyl-, pyridinylphenyl-, furanylphenyl-, thienylphenyl-, and thiazolylphenyl-,
  • substituted heteroarylaryi- such as, for example, substituted pyrimidinylphenyl-, substituted pyrazinylphenyl-, substituted pyridinyiphenyl-, substituted furanylphenyl-, substituted thienylphenyl-, substituted thiazolylphenyl-, and substituted pyrimidinylphenyl,
  • R 8 is selected from the group consisting of: H, -OH, aikyl, aryl, -N(R 10 J 2 and -NR 1 °C(O)R 12 ; each R 9 is independently selected from the group consisting of:halogen, -CN, -NO 2 , -OR 10 , -SR 10 , -N(R 10 J 2 , and R 10 ; each R 10 is independently selected from the group consisting of; H, aikyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkyl aikyl, heterocycloalkyl, heterocycloalkylalkyl, alkylheteroaryl-, alkylaryl-, substituted aikyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloalkyS, substituted cycloalkylal
  • R 1 Ms selected from the group consisting of: F, -OH, -CN, -OR 10 , -NHNR 1 R 10 ,
  • R 12 is selected from the group consisting of: alkyi, aryl, heteroaryi, cycloalkyl, cyctoalkylalkyl, heterocycloalkyl and heterocycloalkylalkyl;
  • R 14 is selected from the group consisting of: aikyl, aryl, heteroaryl, cycloalkyi, cycloalkylalkyl-, heterocycloalkyl, alkylheterocycloalkyl, heterocycloalkylalkyl-, alkylheteroaryl- and alkyiaryl-;
  • R 15 is selected from the group consisting of: H, -OH, alkyl, aryl, heteroaryl, cycioalkyl, cycloalkylalkyl-, heterocycloalkyl and heterocycloalkylalkyl-, alkylheteroaryl- and alkyiaryl-;
  • R 20 represents alkyl;
  • R 23 is selected from the group consisting of: H, alkyl, aryl, cycloalkyl, and cycloalkyialkyl-; each R 26 is independently selected from the group consisting of: H and alkyl; R 28 is alkyl; each R 30 is independently selected from the group consisting of: H, afkyl, and
  • R 42 is selected from the group consisting of: alkyl, aryi, heteroaryl, and cycloalkyi;
  • R 44 is selected from the group consisting of: H, alkyl, cycloaikyl, and cycloalkyialkyl; and Each R 46 is independently selected from the group consisting of: H, alkyl, cycloalkyi, and cycloalkylalkyi;
  • Non-limiting examples of ERK inhibitors are selected from the group consisting of:
  • compositions which comprise at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound and at least one pharmaceutically acceptable carrier.
  • inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. The powders and tablets may be comprised of from about 5 to about 95 percent active ingredient. Suitable solid carriers are known in the art, e.g., magnesium carbonate, magnesium stearate, talc, sugar or lactose.
  • Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration.
  • Examples of pharmaceutically acceptable carriers and methods of manufacture for various compositions may be found in A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18 th Edition, (1990), Mack Publishing Co., Easton, Pennsylvania.
  • Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection or addition of sweeteners and opacifiers for oral solutions, suspensions and emulsions. Liquid form preparations may also include solutions for intranasal administration.
  • Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas, e.g. nitrogen. Also included are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.
  • the compounds of the invention may also be deliverable transdermally.
  • the transdermal compositions can take the form of creams, lotions, aerosols and/or emulsions and can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose.
  • the compounds of this invention may also be delivered subcutaneously.
  • the compound is administered orally or intravenously.
  • delivery methods that are combinations of the above- noted delivery methods, Such methods are typically decided by those skilled in the art.
  • the pharmaceutical preparation is in a unit dosage form.
  • the preparation is subdivided into suitably sized unit doses containing appropriate quantities of the active component, e.g., an effective amount to achieve the desired purpose.
  • the quantity of active compound in a unit dose of preparation may be varied or adjusted from about 1 mg to about 100 mg, preferably from about 1 mg to about 50 mg, more preferably from about 1 mg to about 25 mg, according to the particular application.
  • the actual dosage employed may be varied depending upon the requirements of the patient and the severity of the condition being treated. Determination of the proper dosage regimen for a particular situation is within the skill of the art. For convenience, the total daily dosage may be divided and administered in portions during the day as required.
  • a typical recommended daily dosage regimen for oral administration can range from about 1 mg/day to about 500 mg/day, preferably 1 mg/day to 200 mg/day, in two to four divided doses.
  • kits comprising a therapeutically effective amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound and a pharmaceutically acceptable carrier, vehicle or diluent.
  • kits comprising an amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound and an amount of at least one anticancer therapy and/or anti-cancer agent listed above, wherein the amounts of the two or more ingredients result in desired therapeutic effect.
  • Biosystems API-100 mass spectrometer and Shimadzu SCL-10A LC column Altech platinum C18, 3 micron, 33mm x 7mm ID; gradient flow: 0 min - 10% CH 3 CN, 5 min -
  • NBS N-Bromosuccinimide it: room temperature
  • LiHMDS lithium hexamethyldisilazane
  • HMDS hexamethyldis ⁇ azane
  • Pd/C palladium on carbon
  • PDCI 2 (dppf): [1 ,1 '-bis(diphenylphosphino)ferrocene] dichloropalladium(il) ⁇ mol: micromole TFA: trifluoroacetic acid
  • Pd(PPh 3 ) 4 tetrakis(triphenylphosphine) palladium
  • N-lodosuccinimide (1.26 g, 5.61 mmol, dissolved in 25 ml acetonitrile) was added into a solution of di-tert-buty! 5-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)piperidine-1 ,2-dicarboxylate (3.8 g, 5.61 mmol) in acetonitrile (26 mL). The reaction mixture was stirred at room temperature for 16 hours.
  • N-lodosuccinimide (81 1 mg, 3.6 mmol) was added into a solution of 4 ⁇ 7-[bis-(2- trimethylsilanyl-ethoxymethyl)-amino]-pyrazolo[1 ,5-a]pyrimid!n-5-yl ⁇ - cyclohexanecarboxylic acid methyl ester (1.75 g, 3.27 mmol) in acetonitrile (25 mL). The resulting reaction mixture was stirred at room temperature for 16 hours.
  • N-bromosuccinimide (148 mg, 0.832 mmol) was added to a solution of 4- ⁇ 7-[bis-(2- trimethy!silanyl-ethoxymethyl)-amino]-3-quinolin-3-yl-pyrazolo[1 ,5-a]pyrimidin-5-yl ⁇ - cyciohexanecarboxylic acid methyl ester (500 mg, 0.756 mmol) in acetonitrile (10 ml_). The reaction mixture was stirred at room temperature for 1 hour.
  • Potassium phosphate (3 M -in H 2 O, 0.2 mmoi) was added followed by the addition of 5-chloro-3-(quinolin-3-yl)-N,N- bis((2-(trimethylsily!ethoxy)methyl) pyrazo!o[1 ,5-a]pyrimidin-7-amine (50 mg, 0.09 mmol) in DMF (0.5 ml) and Pd(dppf)CI 2 -CH 2 CI 2 (5 mg, 0.005 mmol). The reaction mixture was heated at 90 °C for 16 h under argon. The solution was cooled to room temperature and concentrated under reduced pressure.
  • Part C To a EtOH (8.0 mL) solution of compound from Part B (1.12 g, 4.1 mmol) was added CuI (39 mg, 0.2 mmol) and followed with Et 3 N (3.5 mL) the resulting mixture was stirred at 70 °C for 3 hours. The reaction mixture was allowed to cool to 25 °C, concentrated to dryness and then dissolved in toluene (3OmL). The solution was washed with 0.1 N HCI (20 mL), saturated NaHCO 3 (20 mL), and brine (30 mL), dried over Na 2 SO 4 .
  • Part D A mixture of the product from Part C (1 equiv), bispinacoiatodiboron (1.2 equiv), PdCI 2 dppf (0.1 equiv), and potassium acetate (3 equiv) in dioxane was heated at 85 °C until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction was allowed to cool to room temperature, filtered through a pad of celite, concentrated to dryness, treated with diethyl ether, filtered through a pad of celite, then concentrated to afford the desired product, 5-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)furo[2,3-b]pyridine.
  • Part A A mixture of 5-bromo-3-iodopyridin-2-ol (1 equiv), copper iodide (3 equiv), palladium catalyst (0.05 equiv), TBAF (1 equiv), triethylamine (3.3 equiv) and alkyne (1 equiv) in toluene was stirred at room temperature until the reaction was deemed complete by TLC analysis. The reaction mixture was diluted with dichloromethane, washed with water, saturated sodium bicarbonate, and brine, dried over sodium bicarbonate, filtered through celite, concentrated, and purified on silica gel to give the product,5-bromo-2-methylfuro[2,3-b]pyridine. Part B:
  • reaction was allowed to cool to room temperature, filtered through a pad of celite, concentrated to dryness, treated with diethyl ether, filtered through a pad of celite, and then concentrated to afford the desired product, 2-methyl-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)furo[2,3- b]pyridine.
  • Part B The 5-bromo-2-(trimethylsilyl)thieno[2,3-b]pyridine from Part A (8.99 g, 31.4 mmol) was dissolved in ethano! (70 mL), followed by addition of K 2 CO 3 (10.85g, 78.5 mmol). Reaction was stirred at 65 °C for 1 hour. The reaction mixture was allowed to cool to 25 °C, concentrated to dryness, and dissolved in EtOAc (150 mL). The solution was washed with H 2 O (80 mL), and brine (15OmL), dried over Na 3 SO 4, filtered, and concentrated to yield white solid, 5-bromothieno[2,3-b]pyridine, 6.57 g (98%).
  • reaction was allowed to cool to room temperature, filtered through a pad of celite, concentrated to dryness, treated with diethyl ether, filtered through a pad of celite, and then concentrated to afford the desired product, 5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)thieno[2,3-b]pyridine.
  • the resulting material was stirred with a mixture of vinyl stannane (2 equiv), palladium catalyst (0.1 equiv), and potassium fluoride (3 equiv) in dioxane at 85 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis.
  • the mixture was allowed to cool to room temperature, diluted with 10% aqueous potassium fluoride, and extracted three times with ethyl acetate. The combined organics were dried over sodium sulfate, filtered, concentrated, and purified on silica gel.
  • the resulting material was stirred with a mixture of HCI in methanol and water at 60 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis.
  • Part D methyl 2-(4-(7-(bis((2-(trimethyisilyl)ethoxy)methyl)amino)-6-bromo-3-(quinolin-3- yl)pyrazolori .5-alpyrimidin-5-yloxy)phenyl)acetate:
  • Part E methyl 2-(4-(7-amino-6-bromo-3-(Quinolin-3-yl)pyrazolo[1 ,5-alpyrimidin-5- yloxy)phenvOacetate: The crude methyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyi)amino)-6-bromo-3-(quinolin- 3-yl)pyrazolo[1,5-a]pyrimidin-5-yloxy)pheny[)acetate from above step D was treated with 50% TFAZH 2 O (3 mL) and stilted at room temperature for 1 h.
  • Part F 4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazoio[l,5-a]pyrimidin-5-yl)benzoic acid:
  • Part C 4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolori .5-aipyrimidin-5-yl)benzoic acid: Compound methyl 4-(6-acetyf-7-amino-3-(6-phenylpyridin-3-yI)pyrazolo[ l,5-a]pyrimtdin-5- yi)benzoate was treated with 2 equivalents of LiOH and stirred at room temperature for 4hrs.
  • Methyl 2-(4-(bromomethyl)phenyl)acetate (1.03 g, 4.24 mmol) was mixed with Pd(PPh3)4 (500 mg, 0.4 mmol, 4,4,4 1 ,4',5,5,5 1 ,5'-octamethyl-2,2 1 -bi(l,3,2-dioxaborolane) (1.27 g, 5 mmol), K2CO3 (1.75 g, 12.7 mmol) and dioxane (20 mL) under inert Argon atmoshphere. The resulting mixture was heated at 80 °C and stirred over night.
  • Pd(PPh3)4 500 mg, 0.4 mmol, 4,4,4 1 ,4',5,5,5 1 ,5'-octamethyl-2,2 1 -bi(l,3,2-dioxaborolane)
  • K2CO3 (1.75 g, 12.7 mmol
  • dioxane 20 mL
  • Part B 5-(4-(methylthio)benzyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5- aipyrimidin-7-amine:
  • Part F ⁇ -bromo-5-(4-(methylsulfonyl)benzyl)-3-(quinoiin-3-yl)-N,N-bis((2-
  • Ttile compound 6-bromo-5-(4-(methylsulfonyl)benzyl)-3-(quinolin-3-yl)pyrazolo[1,5- a]pyrimidin-7-amine was prepared with the same deprotection condition described in example I Part F.
  • HPLC-MS t R 1.49 min (UV 254 nm ); mass calculated for formula C 23 Hj 8 BrN 5 O 2 S 507.0, observed LCMS m/z 5O8.O(M+H).
  • step E The crude from above step E was treated with 50% TFA/H2O (3 mL) and stirred at room temperature for 1 h. Solvent was removed to yield thick oil, methyl 4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-6-(l-ethoxyvinyl)-3-(6-phenylpyridin-3-yl)pyrazolo[1,5- a]pyrimidin-5-yioxy)benzoate.
  • HPLC-MS IR 1.84 min (UV 254 nm ); mass calculated for formula C 27 H 21 N 5 O 4 479.2, observed LCMS m/z 480.2 (M+H).

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Pulmonology (AREA)
  • Hematology (AREA)
  • Oncology (AREA)
  • Transplantation (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The present invention provides methods for inhibiting mTOR using pyrazolo[1,5-a]pyrimidine compounds and methods of treatment, prevention, inhibition, or amelioration of one or more diseases associated with mTOR using such compounds.

Description

PYRAZOLO [1 , 5-A] PYRIMIDINE DERIVATIVES AS MTOR INHIBITORS
FIELD OF THE INVENTION
This invention is directed to pyrazolo pyrimidine derivatives as inhibitors of mammalian Target Of Rapamycin (mTOR) kinase, which are also known as FRAP, RAFT, RAPT or SEP and as a result have a role in controlling cell cycle progression and mitogenic signals involved in cell growth and proliferation and as a result are useful in the treatment of cancer.
BACKGROUND OF THE INVENTION
The mammalian target of rapamycin (mTOR) is a central regulator of cell growth and proliferation and plays a gate keeper role in the control of cell cycle progression and mediates mitogenic signals from P13K/AKT through to the downstream targets S6K1 and 4E-BP1 and to Ser 473 on AKT. Recently it has been shown that mTOR exists in two complexes, raptor-mTOR complex (mTORCI ), a rapamycin-sensitive complex, signaling to S6K1 and 4E-BP1 and rictor-mTOR complex (mTORC2), a rapamycin-insensitive complex that signals to AKT. Although the precise mechanism by which rapamycin inhibits mTOR function is not well understood, rapamycin partially inhibits mTOR function through mTORCI . It has been found that mTORC2 is involved in the regulation of cell survival and actin cytoskeletal organization in a rapamycin-independent manner, and inhibition of mTOR through inhibition of mTORCI and mTORC2 is probably important for antitumor activity and better efficacy.
US 2007/0112005 describes the fused bicyclic mTOR inhibitors useful in treatment of cancer.
WO 2007/087395 describes unsaturated mTOR inhibitors useful in treatment of cancer. WO-2008/012326 describes 2,4-substituted quinozolines as lipid kinase inhibitors useful in treatment of P13K -related diseases, such as proliferative diseases, inflammatory diseases, obstructive airways disorder and transplant related diseases.
WO 2006/090169 describes 2,4-diamineo-pyrido-pyrmidine derivatives and their use as mTOR inhibitors.
WO 2007/066099 describes pyrimidine derivatives useful as mTOR kinase inhibitors for anticancer and various other therapeutic treatments involving mTOR kinase.
WO 2007044813 describes pyridopyrimidinone inhibitors of PI3Ka protein kinase for anticancer or other PI3Ka protein kinase related diseases.
US 2005/0222171 , WO 2005/070431 , WO 2007/0570431 and WO 2007/009773 describe pyrazolo[1 ,5 a]pyrimidin-7-yl amine derivatives to treat protein kinase dependent diseases.
US 2002/0041880 describes pyrazolo[1 ,5 a]pyrimidin-7-yl derivatives to inhibit kinase insert domain-containing receptor to block angiogenesis.
WO 1998/003510 describes pyrazolo[1 ,5 a]pyrimidin-7-yl derivatives to treat corticotrophin releasing factor dependent diseases.
WO 2003/091256 describes pyrazolo[1 ,5-a]pyrimidine derivatives to treat NAD(P)H oxidase dependent diseases.
WO 2007/044449, WO 2005/077954, WO 2004/022560 and WO 2004/022561 describe pyrazolopyrimidine derivatives as cyclin-dependent kinase dependent diseases.
WO 2004/106341 describes pyrazolopyrimidine derivatives as fungicides There is need in the art for small molecule inhibitors of mTOR kinase that block signaling through mTORCI and mT0RC2 as a potential anticancer treatment or a treatment for other cell proliferative disorders.
SUMMARY OF THE INVENTION
In its many embodiments, the present invention discloses novel compounds having mTOR inhibitory activity, methods of preparing such compounds, pharmaceutical compositions comprising one or more of such compounds, methods of treatment or prevention of one or more diseases associated with mTOR by administering one or more of such compounds or pharmaceutical compositions, the compound being represented by the general Formula I:
Figure imgf000004_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is independently selected from the group consisting of heterocycloalkyl, heterocycloalkylalkyl, spiroheterocycloalkyl, heterocyclenyl, -NR3R4, cycloalkyl, heteroaryl, aryl, alkyl, alkynyl, heterocyclenylalkyl, cycloalkylalkyl, heteroarylalkyl, heteroarylalkynyl, spiroheterocycloalkylalkyl, -N-heteroaryl, -alkyl-NH-heterocyclyl and arylalkyl, wherein each of said heterocycloalkyl, heterocycloalkylalkyl, spiroheterocycloalkyl, heterocyclenyl, cycloalkyl, heteroaryl, aryl, alkyl, alkynyl, heterocyclenylalkyl, cycloalkylalkyl, heteroarylalkyl, heteroarylalkynyl, -N-heteroaryl and arylalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X; X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, -alkyl- C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, -alkyl(CO)NS(O)2- cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SO2-cycloalkyl, -alkyl(CO)NS(O)2-alkyl, - alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2-cycloalkyl, -CO-CO2H, - C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl;
R3 is cycloalkyl or heteroaryl, wherein each of said cycloalkyl or heteroaryl can be unsubstituted or substituted with one or more moieties independently selected from the group consisting of X; and R4 is H.
In another embodiment, the present invention includes a method of inhibiting mammalian Target Of Rapamycin in a patient, wherein the method comprises administering a therapeutically effective amount of at least one compound of the structural Formula I. In another embodiment, the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, wherein the method comprises administering a therapeutically effective amount of at least one compound represented by the structural Formula I.
In another embodiment, the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, wherein the method comprises administering a therapeutically effective amount of at least one compound represented by the structural Formula I, to a patient in need thereof.
In another embodiment, the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, and disorders commonly occurring in connection with transplantation, diseases that respond to inhibition of mammalian Target Of Rapamycin, wherein the method comprises administering a therapeutically effective amount of compound represented by the structural Formula I, to a patient in need of such treatment. In another embodiment, the present invention includes a method of treatment of a disease, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I.
In another embodiment, the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, and disorders commonly occurring in connection with transplantation, diseases that respond to inhibition of mammalian Target Of Rapamycin, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I, wherein the disease is a proliferative disease, autoimmune disease, viral disease, fungal disease, neurological/neurodegenerative disorder, arthritis, inflammation, neuronal, alopecia or cardiovascular disease.
In another embodiment, the present invention includes a method of treatment of a disease, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I, wherein the disease is a proliferative disease.
In another embodiment, the present invention includes a method of treatment of a proliferative disease, wherein the method comprises administering a therapeutically effective amount of the compound represented by the structural Formula I, wherein the proliferative disease is selected from the group consisting of: cancer of the bladder, breast, colon, kidney, liver, lung, small cell lung cancer, non- small cell lung cancer, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma, Burkett's lymphoma; acute and chronic myelogenous leukemia, myelodysplastic syndrome, promyelocytic leukemia; fibrosarcoma, rhabdomyosarcoma; astrocytoma, neuroblastoma, glioma and schwannomas; melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderma pigmentosum, keratoctanthoma, thyroid follicular cancer, endometrial cancer, gastrointestinal tract cancer and Kaposi's sarcoma.
In another embodiment, the present invention includes a method of treatment of a proliferative disease comprising administering a therapeutically effective amount of the compound represented by the structural Formula I, further comprising treatment with radiation therapy.
In another embodiment, the present invention includes a method of inhibiting a mammalian Target Of Rapamycin, wherein the method comprises administering a therapeutically effective amount of a compound represented by the structural Formula I.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000007_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is heterocycloalkyl, wherein said heterocycloalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-aikyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl; R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl. In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000008_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo; R1 is heterocyclenyl, wherein said heterocyclenyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SOa-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CHa)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000009_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is heteroaryl, wherein said heteroaryl can be unsubstituted or substituted with one or moieties independently selected from the group X; X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl - C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2) -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000010_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is cycloalkyl, wherein said cycloalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl-
C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000011_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein: R is independently selected from the group consisting of halo, hydroxyl, amino,
-CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is cycloalkenyl, wherein said cycloalkenyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3) -N-alkyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heteroeycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heteroeycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000012_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is aryl, wherein said aryl can be unsubstituted or substituted with one or moieties independently selected from the group X; X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heteroeycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl (CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-eycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heteroeycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl; R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000013_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is alkyl, wherein said alkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl-
C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000014_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein: R is independently selected from the group consisting of halo, hydroxyl, amino,
-CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is alkynyl, wherein said alkynyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(0)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000015_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(C1-C6)alkyl, alkoxy, -C(=O)alkyl, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl and halo;
R1 is spiroheterocycloalkyl, wherein said spiroheterocycloalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2, -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-alkyl, -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000016_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R1 is independently selected from the group consisting of:
Figure imgf000016_0002
Figure imgf000017_0001
Figure imgf000018_0001
Figure imgf000019_0001
Figure imgf000020_0001
R2 is quinolinyl, 1 -methyl-pyrazolyl, naphtyl, methoxy-naphtyl, cyano-phenyl, methoxy-phenyl, phenoxy-phenyl, biphenyl, chloro-pyridyl, methoxy-pyridyl, bromo- methyl-pyrazolyl, hydroxyl-methoxy-phenyl, di-methoxy-phenyl, 1 H-indazolyl, fluoro- hydroxy-phenyl, chloro-quinolyl, morpholino-pyridyl-, 4-(isobutyronitrilo)-phenyl, bromo-quinolyl, 4-trifluoromethyl-phenyl-, benzyl -pyrazolyl, fluoro-pyridyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000020_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein: R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1 -methyl-pyrazolyl, and thienyl; R1 is independently selected from the group consisting of:
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
Figure imgf000024_0001
Figure imgf000025_0001
R2 is indazolyl, wherein said indazolyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000025_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R1 is independently selected from the group consisting of:
Figure imgf000026_0001
Figure imgf000027_0001
Figure imgf000028_0001
Figure imgf000029_0001
Figure imgf000030_0001
R2 is phenyl, wherein said phenyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo, alkyl, alkoxy, -CN, hydroxyl, aryl and heteroaryl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000030_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R1 is independently selected from the group consisting of:
Figure imgf000031_0001
Figure imgf000032_0001
Figure imgf000033_0001
Figure imgf000034_0001
Figure imgf000035_0001
R2 is naphthyl, wherein said naphthyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000035_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R1 is independently selected from the group consisting of:
Figure imgf000036_0001
Figure imgf000038_0001
O=
Figure imgf000039_0001
Figure imgf000040_0001
R2 is quinolinyl, wherein said quinolinyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000040_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R1 is independently selected from the group consisting of:
Figure imgf000041_0001
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
R2 is pyrazolyl, wherein said pyrazolyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000045_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of bromo, chloro, -CN, H, methyl, acetyl, pyridyl, phenyl, 1-methyl-pyrazolyl, and thienyl; R1 is independently selected from the group consisting of:
Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
R2 is pyridinyl, wherein said pyridinyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo, alkyl, heterocyclyl, heteroaryl, aryl and alkoxy.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000050_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is halo;
R1 is independently selected from the group consisting of:
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
O
Figure imgf000054_0001
Figure imgf000055_0001
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, heteroaryl and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000055_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is thienyl; R1 is independently selected from the group consisting of:
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3^CN, trifluromethyl, difluromethyl, monofluromethyl, heterocycloalkyl, heteroaryl and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000060_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is pyrazolyl, wherein said pyrazolyl can be unsubstituted or substituted with alkyl;
R1 is independently selected from the group consisting of:
Figure imgf000061_0001
Figure imgf000062_0001
Figure imgf000063_0001
Figure imgf000064_0001
Figure imgf000065_0001
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000065_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is pyridinyl; R1 is independently selected from the group consisting of:
Figure imgf000066_0001
Figure imgf000067_0001
Figure imgf000068_0001
Figure imgf000069_0001
Figure imgf000070_0001
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyljieterocycloalkyl, heteroaryl and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000070_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is methyl; R1 is independently selected from the group consisting of:
Figure imgf000071_0001
Figure imgf000072_0001
Figure imgf000073_0001
Figure imgf000074_0001
Figure imgf000075_0001
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CHa)2CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
In another embodiment, the present invention relating to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000075_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is phenyl, wherein said phenyl can be unsubstituted or substituted with one or more moieties which can be the same or different each moiety being independently selected from the group consisting of halo or alkyl;
R1 is independently selected from the group consisting of:
Figure imgf000076_0001
Figure imgf000077_0001
Figure imgf000078_0001
Figure imgf000079_0001
Figure imgf000080_0001
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000080_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is acetyl; R1 is independently selected from the group consisting of:
Figure imgf000081_0001
Figure imgf000082_0001
Figure imgf000083_0001
Figure imgf000084_0001
Figure imgf000085_0001
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyljieterocycloalkyl, heteroaryl and arylalkyl.
In another embodiment, the present invention relates to the foregoing method of using a compound represented by the structural Formula I:
Figure imgf000085_0002
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is -CN; R1 is independently selected from the group consisting of:
Figure imgf000086_0001
Figure imgf000087_0001
Figure imgf000088_0001
Figure imgf000089_0001
Figure imgf000090_0001
and
R2 is hβteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, heteroaryl and arylalkyl.
Non-limiting examples of the compounds of the present invention include.
Figure imgf000090_0002
Figure imgf000091_0001
Figure imgf000092_0001
Figure imgf000093_0001
93
Figure imgf000094_0001
Figure imgf000095_0001
95
Figure imgf000096_0001
Figure imgf000097_0001
Figure imgf000098_0001
Figure imgf000099_0001
Figure imgf000100_0001
Figure imgf000101_0001
Figure imgf000102_0001
Figure imgf000103_0001
Figure imgf000104_0001
Figure imgf000105_0001
Figure imgf000106_0001
Figure imgf000107_0001
Figure imgf000108_0001
Figure imgf000109_0001
Figure imgf000110_0001
Figure imgf000111_0001
Figure imgf000112_0001
Figure imgf000113_0001
Figure imgf000114_0001
114
Figure imgf000115_0001
Figure imgf000116_0001
Figure imgf000117_0001
Figure imgf000118_0001
118
Figure imgf000119_0001
Figure imgf000120_0001
Figure imgf000121_0001
Figure imgf000122_0001
Figure imgf000123_0001
Figure imgf000124_0001
124
Figure imgf000125_0001
Figure imgf000126_0001
Figure imgf000127_0001
Figure imgf000128_0001
Figure imgf000129_0001
Figure imgf000130_0001
Figure imgf000131_0001
Figure imgf000132_0001
Figure imgf000133_0001
Figure imgf000134_0001
Figure imgf000135_0001

Figure imgf000136_0001
Figure imgf000137_0001
Figure imgf000138_0001
Figure imgf000139_0001

Figure imgf000140_0001
Figure imgf000141_0001
Figure imgf000142_0001
Figure imgf000143_0001
or pharmaceutically acceptable salt, ester, solvate or prodrug thereof.
In another embodiment, the present invention relates to a compound represented by the structural Formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof, in purified form.
In another embodiment, the present invention relates to a compound represented by the structural Formula I or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof, in isolated form.
In another embodiment, the present invention includes a composition comprising a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, in combination with at least one pharmaceutically acceptable carrier.
In another embodiment, the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, said method comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above, or a pharmaceutically acceptable salt, solvate, ester or prodrug of the compound, to a patient in need thereof.
In another embodiment, the present invention includes a composition comprising a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof, and an anti-cancer agent.
In another embodiment, the present invention includes a composition comprising a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof, and an anti-neoplastic, anti-tumor, anti-angiogenic, or chemotherapeutic agent.
In another embodiment, the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, and disorders commonly occurring in connection with transplantation, diseases that respond to inhibition of nnTOR, comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
In another embodiment, the present invention includes a method of treatment of a proliferative disease, autoimmune disease, viral disease, fungal disease, neurological/neurodegenerative disorder, arthritis, inflammation, neuronal, alopecia or cardiovascular disease comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
In another embodiment, the present invention includes a method of treatment of a proliferative disease.
In another embodiment, the present invention includes a method of treatment of a proliferative disease, wherein the proliferative disease is selected from the group consisting of: cancer of the bladder, breast, colon, kidney, liver, lung, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin; small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma, Burkett's lymphoma; acute and chronic myelogenous leukemia, myelodysplastic syndrome, promyelocyte leukemia; fibrosarcoma, rhabdomyosarcoma; astrocytoma, neuroblastoma, glioma and schwannomas; melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderma pigmentosum, keratoctanthoma, thyroid follicular cancer, endometrial cancer, gastrointestinal tract cancer and Kaposi's sarcoma.
In another embodiment, the present invention includes a method of treatment of hamartoma syndromes, transplant rejection, bowel disorders, inflammatory bowel disease, multiple sclerosis, immunosuppression, immune tolerance, autoimmune diseases, inflammation, bone loss, rheumatoid arthritis, restinosis, cardiac allograft vasculopathy, psoriasis, ocular conditions such as dry eye, hepatic fibrosis, hepatic necrosis, beta-thalassaemia, comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
In another embodiment, the present invention includes a method of treatment of cancer of the bladder, breast, colon, kidney, liver, lung, small cell lung cancer, non- small cell lung cancer, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma, Burkett's lymphoma; acute and chronic myelogenous leukemia, myelodysplastic syndrome, promyelocytic leukemia; fibrosarcoma, rhabdomyosarcoma; astrocytoma, neuroblastoma, glioma and schwannomas; melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderma pigmentosum, keratoctanthoma, thyroid follicular cancer, endometrial cancer, gastrointestinal tract cancer and Kaposi's sarcoma comprising administering a therapeutically effective amount of at least one compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof, further comprising treatment with radiation therapy to a patient in need of such treatment.
In another embodiment, the present invention includes a method of treating a disease by inhibiting a mTOR, comprising administering to a patient in need of such treatment an amount of a first compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof; and an amount of at least one second compound, said second compound being an anti-cancer agent; wherein said anti-cancer agent is selected from the group consisting of Adriamycin, Altretamine, Amidox, Aminoglutethimide, Amsacrine, Anastrazole, Antibodies to EGFR, 3-AP, Aphidicolon, Ara-C, Arsenic trioxide, L-Asparaginase, Bevacizumab, Bleomycin, BMS 214662, Bortezomib, Busulfan, Campath, Camptostar, Capecitabine, Carboplatin, Carmustine, Centrosome associated protein E ("CENP-E") inhibitors, Cetuximab, Cladribine, Chlorambucil, Chlormethine, Chlorotrianisene, Cisplatin, Clofarabine, cyclophosphamide, Cytarabine, a Cytostatic agent, Cytoxan, Dacarbazine, Dactinomycin, Daunorubicin, Dasatinib, Deforolimus (described in PCT publication No. 2003/064383), Deoxycoformycin, Didox, Diethylstilbestrol, Docetaxel, Doxorubicin, Dromostanolone, Droloxafine, Epirubicin, Epothilones, ERK inhibitors, Erlotinib, Etoposide, 17α-Ethinylestradiol, Estramustine, Exemestane, Floxuridine, Fludarabine, Fludarabine phosphate, 5-Fluorouracil, Fluoxymesterone, Flutamide, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab ozogamcicin, Goserelin, GSK- 923295, Hexamethylmelamine, Hydroxyprogesterone, Hydroxyurea, lbritumomab Tiuxetan, Idarubicin, Ifosfamide, lmatinib mesylate, Intron, Irinotecan, ispinesib, KSP inhibitors, L778,123, Lapatinib, Leucovirin, Leuprolide, Lerozole, Letrazole, Levamisole, Liposomal Doxorubicin, Liposomal, Lomustine, Lonafarnib,
Medroxyprogesteroneacetate, Megestrolacetate, Melphalan, 6-Mercaptopurine, Methoxtrexate, Methylprednisolone, Methyltestosterone, Mithramycin, Mitomycin-C, Mitotane, Mitoxantrone, Navelbene, Nilotinib, Oxaliplatin, Paclitaxel, Panitubimab, Pentostatin, Pipobroman, Porfimer, Prednisolone, Prednisone propionate, Procarbazine, Reloxafine, Rituximab, Satriplatin, SB-743921 , SmH , Sorafinib, Streptozocin, Sunitinib, Tamoxifen, Taxotere, Taxol, Temozolomide, Teniposide, Testolactone, Testosterone, Tezacitabine, 6-Thioguanine, Thiotepa, Tipifamib, Topotecan, Toremifene, Tositumomab, Trastuzumab, Triamcinolone, Triapine, Triethylenemelamine, Triethylenethiophosphoramine, Trimidox, Uracil mustard, Vinblastine, Vincristine, Vindesine, and Vinorelbine; wherein the amounts of the first compound and said second compound result in a therapeutic effect.
In another embodiment, the present invention includes a method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, said method comprising administering a therapeutically effective amount of at least one compound of Formula I, or a pharmaceutically acceptable salt, solvate, ester or prodrug of the compound, to a patient in need thereof.
In another embodiment, the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, diseases related to transplant rejection and diseases that respond to inhibition of mTOR, comprising administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
In another embodiment, the present invention includes a method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, diseases related to transplant rejection and diseases that respond to inhibition of mTOR, comprising administering a therapeutically effective amount of at least one compound of Formula I or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
In another embodiment, the present invention includes a compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof exhibiting mTOR inhibition which is at least five-fold the inhibition of CDK2 or CHK-1 by said compound.
In another embodiment, the present invention includes a compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof exhibiting mTOR inhibition which is at least ten-fold the inhibition of CDK2 or CHK-1 by said compound. In another embodiment, the present invention includes a compound from the group of compounds listed above or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof exhibiting mTOR inhibition which is at least fifty-fold the inhibition of CDK2 or CHK-1 by said compound. The compounds of this invention can be used to inhibit the following kinases:
ABL1 , ABL2, AFK, ALK, AMPK, ATM, ATR, Aurora A, Aurora B, AxI, BCKDK, BLK, BMPR1 B, BMX, Brk, BRSK1 , BTK, CaM-Klalpha, CaM-Kllalpha, CaM-KIV, CaM- KKalpha, CaM-KKbeta, CCDPK, CCRK, CDK1 , CDK11 , CDK2, CDK4, CDK5, CDK6, CDK7, CDK9, Chaki , CHK1 , CHK2, CK1 alpha, CK1 delta, CDkI epsilon, CDK2 beta, CLK1 , CSF1 R, Csk, DAPK1 , DAPK2, DAPK3, DCAMKL1 , DNA-PK, DYRK1 A,
DYRK1 B, DYRK2, DYRK3, eEF2K, Eg3, EGFR, EIF2AK2, EphA2, EphA3, EphA4, EphA8, EphB1, EphB2, EphB3, EphB5, ErbB2, FAK, Fer, Fes, FGFR1 , FGFR3, FGFR4, Fgr, FLT1 , FLT3, FLT4, Fyn, GRK-1 , GRK-2, GRk-3, GRK-4, GRK-5, GRK-6, GSK-3alpha, GSK-3beta, HCK, HIPK2, HIPK3, HRI, ICK, IGF1 R, IKK-alpha, IKK-beta, IKK-epsilon, ILK, InsR, IPL1 , IRAKI , IRAK4, ITK, JAK1 , JAK2, JAK3, JNK1 , JNK2, JNK3, KDR, KIS, Kit, KSR1 , Lck, LIMK1 , LIMK2, LKB1 , LOK, Lyn, MAP2K1 , MAP2K2, MAP2K3, MAP2K4, MAP2K6, MAP2K7, MAP3K1 , MAP3K11 , MAP3K14, MAP3K5, MAP3K7, MAP3K8, MAP4K1 , MAP4K2, MAP4K4, MAPK1 , MAPK10, MAPK11 , MAPK12, MAPK13, MAPK14, MAPK3, MAPK4, MAPK6, MAPK7, MAPK8, MAPK9, MAPKAPK2, Mer, Met, MHCK, MLCK, Mnk1 , Mnk2, MOS, MRCKa, MST1 , MST3, NDR1 , NDR2, NEK1 , NEK2, NEK6, NEK9, NLK, NuaK1 , p37, p38, p70S6K, p70S6Kb, PAK1 , PAK2, PAK2, PAK3, PAK5, PAK6, PASK, P-CIP2, PCTAIRE1 , PDGFR alpha, PDGFR beta, PDHK1 , PDHK2, PDHK3, PDHK4, PDK-1 , PDK-2, PHK, PIK3CA, PIK3CB, PIK3CD, PIK3CG, Pim-1 , PKA alpha, PKB beta, PKC alpha, PKC beta, PKC delta, PKC epsilon, PKC eta, PKC gamma, PKC iota, PKC theta, PKC zeta, PKD1 , PKD2, PKD3, PKG1/cGK-l, PKG1/cGK-ll, PKN1 , PLK1 , PLK2, PLK3, PRP4, PYK2, RAF1 , Ret, ROCK1 , ROCK2, Ron, RPL10, RSK-1 , RSK-2, RSK-3, RSK-5, SDK1 , SIK, Sky, Src, STLK3, Syk, TBK1 , Tec, TESK1 , TESK2, TGFbRI , TGFbR2, Tie1 , Tie2, Titin kinase, TNK2, TRKA, TRB, tropomyosin kinase, TSSK3, TXK, Tyk2, VRK1 , Wee1 , Wnk1 , Yes, and ZAP70. As used above, and throughout this disclosure, the following terms, unless otherwise indicated, shall be understood to have the following meanings, including any possible substitutions of the stated groups or moieties: "Patient" includes both human and animals.
"Mammal" means humans and other mammalian animals. "Alkyl" means an aliphatic hydrocarbon group which may be straight or branched and comprising about 1 to about 20 carbon atoms in the chain. Preferred alkyl groups contain about 1 to about 12 carbon atoms in the chain. More preferred alkyl groups contain about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. "Lower alkyl" means a group having about 1 to about 6 carbon atoms in the chain which may be straight or branched. "Alkyl" may be unsubstituted or optionally substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of halo, alkyl, aryl, cycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, oxime (e.g., =N-OH), - NH(alkyl), -NH(cycloalkyl), -N(alkyl)2, -O-C(O)-alkyl, -O-C(O)-aryl, -O-C(O)-cycloalkyl, carboxy and -C(O)O-alkyl. Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl and t-butyl. "Alkenyl" means an aliphatic hydrocarbon group containing at least one carbon- carbon double bond and which may be straight or branched and comprising about 2 to about 15 carbon atoms in the chain. Preferred alkenyl groups have about 2 to about 12 carbon atoms in the chain; and more preferably about 2 to about 6 carbon atoms in the chain. Branched means that one or more, lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkenyl chain. "Lower alkenyl" means about 2 to about 6 carbon atoms in the chain which may be straight or branched. "Alkenyl" may be unsubstituted or optionally substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of halo, alkyl. aryl, cycloalkyl, cyano, alkoxy and -S(alkyl). Non-limiting examples of suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut- 2-enyl, n-pentenyl, octenyl and decenyl. "Alkylene" means a difunctional group obtained by removal of a hydrogen atom from an alkyl group that is defined above. Non-limiting examples of alkylene include methylene, ethylene and propylene.
"Alkynyl" means an aliphatic hydrocarbon group containing at least one carbon- carbon triple bond and which may be straight or branched and comprising about 2 to about 15 carbon atoms in the chain. Preferred alkynyl groups have about 2 to about 12 carbon atoms in the chain; and more preferably about 2 to about 4 carbon atoms in the chain. Branched means that one or more, lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkynyl chain. "Lower alkynyl" means about 2 to about 6 carbon atoms in the chain which may be straight or branched. Non-limiting examples of suitable alkynyl groups include ethynyl, propynyl, 2-butynyl and 3- methylbutynyl. "Alkynyl" may be unsubstituted or optionally substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of alkyl, aryl and cycloalkyl. "Aryl" means an aromatic monocyclic or multicyclic ring system comprising about 6 to about 14 carbon atoms, preferably about 6 to about 10 carbon atoms. The aryl group can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined herein. Non-limiting examples of suitable aryl groups include phenyl and naphthyl. "Bridged cyclic ring" is a hydrocarbon ring such as cycloalkyl, cyclenyl, or aryl or heteroatom containing ring such as, heterocyclyl, heterocyclenyl, or heteroaryl as described herein, that contains a bridge, which is a valence bond or an atom or an unbranched chain of atoms connecting two different parts of the ring. The two tertiary carbon atoms connected through the bridge are termed "bridgeheads". "Heteroaryl" means an aromatic monocyclic or multicyclic ring system comprising about 5 to about 14 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the ring atoms is an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. Preferred heteroaryls contain about 5 to about 6 ring atoms. The "heteroaryl" can be optionally substituted by one or more "ring system substituents" which may be the same or different, and are as defined herein. The prefix aza, oxa or thia before the heteroaryl root name means that at least a nitrogen, oxygen or sulfur atom respectively, is present as a ring atom. A nitrogen atom of a heteroaryl can be optionally oxidized to the corresponding N-oxide. "Heteroaryl" may also include a heteroaryl as defined above fused to an aryl as defined above. Non-limiting examples of suitable heteroaryls include pyridyl, pyrazinyl, furanyl, thienyl, pyrimidinyl, pyridone (including N-substituted pyridones), isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl, pyrazolyl, triazolyl, 1 ,2,4- thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl, imidazo[1 ,2- a]pyridinyl, imidazo[2,1 -b]thiazolyl, benzofurazanyl, indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl, imidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl, pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl, 1 ,2,4-triazinyl, benzothiazolyl and the like. The term "heteroaryl" also refers to partially saturated heteroaryl moieties such as, for example, tetrahydroisoquinolyl, tetrahydroquinolyl and the like.
"Aralkyl" or "arylalkyl" means an aryl-alkyl- group in which the aryl and alkyl are as previously described. Preferred aralkyls comprise a lower alkyl group. Non-limiting examples of suitable aralkyl groups include benzyl, 2-phenethyl and naphthalenylmethyl. The bond to the parent moiety is through the alkyl.
"Alkylaryl" means an alkyl-aryl- group in which the alkyl and aryl are as previously described. Preferred alkylaryls comprise a lower alkyl group. Non-limiting example of a suitable alkylaryl group is tolyl. The bond to the parent moiety is through the aryl.
"Cycloalkyl" means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7 ring atoms. The cycloalkyl can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined above. Non-limiting examples of suitable monocyclic cycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Non-limiting examples of suitable multicyclic cycloalkyls include 1 -decalinyl, norbomyl, adamantyl and the like.
"Cycloalkylalkyl" means a cycloalkyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core. Non-limiting examples of suitable cycloalkylalkyls include cyclohexylmethyl, adamantylmethyl and the like. "Cycloalkenyl" means a non-aromatic mono or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms which contain at least one carbon-carbon double bond. Preferred cycloaikenyl rings contain about 5 to about 7 ring atoms. The cycloalkenyl can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined above. Non-limiting examples of suitable monocyclic cycioalkenyls include cyclopentenyi, cyclohexenyl, cyclohepta-1 ,3-dienyl, and the like. Non-iimiting example of a suitable multicyclic cycloalkenyl is norbornylenyl.
"Cycloalkenylalkyl" means a cycloalkenyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core. Non-limiting examples of suitable cycloalkenyialkyls include cyclopentenylmethyl, cyclohexenylmethyl and the like.
"Halogen" means fluorine, chlorine, bromine, or iodine. Preferred are fluorine, chlorine and bromine.
"Ring system substituent" means a substituent attached to an aromatic or non- aromatic ring system which, for example, replaces an available hydrogen on the ring system. Ring system substituents may be the same or different, each being independently selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryi, araikyl, alkylaryl, heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkyl heteroaryl, hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo, nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyi, aralkoxycarbonyl, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio, cycloalkyl, heterocyclyl, amide, -CHO, -O-C(O)-alkyl, -O-C(O)-aryl, - O-C(O)-cycloalkyl, -C(=N-CN)-NH2l -C(=NH)-NH2, -C(=NH)-NH(alkyl), oxime (e.g., =N-OH), YiY2N-, Y1Y2N-^kVl-, Y1Y2NC(O)-, Y1Y2NSO2- and -SO2NY1Y2, wherein Y1 and Y2 can be the same or different and are independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyi, and araikyl. "Ring system substituent" may also mean a single moiety which simultaneously replaces two available hydrogen on two adjacent carbon atoms (one H on each carbon) on a ring system. Examples of such moiety are methylene dioxy, ethyl enedioxy, -C(CH3J2- and the like which form moieties such as, for example:
Figure imgf000153_0001
"Heteroarylalky!" means a heteroary! moiety as defined above linked via an alkyl moiety (defined above) to a parent core. Non-limiting examples of suitable heteroaryis include 2-pyridinylmethyl, quinolinylmethy! and the like. "Heterocyciyl" means a non-aromatic saturated monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the atoms in the ring system is an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and/or sulfur atoms present in the ring system. Preferred heterocyclyls contain about 5 to about 6 ring atoms. The prefix aza, oxa or thia before the heterocyciyl root name means that at least a nitrogen, oxygen or sulfur atom respectively is present as a ring atom. Any -NH in a heterocyciyl ring may exist protected such as, for example, as an -N(Boc), -N(CBz), -N(Tos) group and the like; such protections are also considered part of this invention. The heterocyciyl can be optionally substituted by one or more "ring system substituents" which may be the same or different, and are as defined herein. The nitrogen or sulfur atom of the heterocyciyl can be optionally oxidized to the corresponding N-oxide, S-oxide or S1S- dioxide. Non-limiting examples of suitable monocyclic heterocyciyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyi, thiazolidinyl, 1 ,4- dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, lactam, lactone, and the like.
"Heterocyciyl" may also mean a single moiety (e.g., carbonyl) which simultaneously replaces two available hydrogen on the same carbon atom on a ring system. Example of such moiety is pyrrolidone:
Figure imgf000153_0002
. "Heterocyclylalkyl" means a heterocyclyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core. Non-limiting examples of suitable heterocyclylaikyls include piperidinylmethyl, piperazinylmethyl and the like.
"Heterocyclenyl" means a non-aromatic monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the atoms in the ring system is an element other than carbon, for example nitrogen, oxygen or sulfur atom, alone or in combination, and which contains at least one carbon-carbon double bond or carbon-nitrogen double bond. There are no adjacent oxygen and/or sulfur atoms present in the ring system. Preferred heterocycienyl rings contain about 5 to about 6 ring atoms. The prefix aza, oxa or thia before the heterocyclenyl root name means that at least a nitrogen, oxygen or sulfur atom respectively is present as a ring atom. The heterocycienyl can be optionally substituted by one or more ring system substituents, wherein "ring system substituent" is as defined above. The nitrogen or sulfur atom of the heterocyclenyl can be optionally oxidized to the corresponding N-oxide, S-oxide or S,S-dioxide. Non- limiting examples of suitable heterocyclenyl groups include 1 ,2,3,4- tetrahydropyridinyl, 1 ,2-dihydropyridinyl, 1 ,4-dihydropyridinyl, 1 ,2,3,6- tetrahydropyridinyl, 1 ,4,5,6-tetrahydropyrimidinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2- imidazolinyl, 2-pyrazoiinyl, dihydroimidazolyl, dihydrooxazolyi, dihydrooxadiazolyl, dihydrothiazolyl, 3,4-dihydro~2H-pyranyS, dihydrofuranyl, fluorodihydrofuranyi, 7- oxabicyclo[2.2.1]heptenyl, dihydrothiophenyi, dihydrothiopyranyl, and the like. "Heterocyclenyl" may also mean a single moiety (e.g., carbonyl) which simultaneously replaces two available hydrogen on the same carbon atom on a ring system. Example of such moiety is pyrrolidinone:
Figure imgf000154_0001
"Heterocyclenylalkyl" means a heterocyclenyl moiety as defined above linked via an alkyl moiety (defined above) to a parent core. It should be noted that in hetero-atom containing ring systems of this invention, there are no hydroxyl groups on carbon atoms adjacent to a N, O or S, as well as there are no N or S groups on carbon adjacent to another heteroatom. Thus, for example, in the ring:
Figure imgf000155_0001
there is no -OH attached directly to carbons marked 2 and 5.
It should also be noted that tautomeric forms such as, for example, the moieties:
Figure imgf000155_0002
are considered equivalent in certain embodiments of this invention.
"Alkynylalkyl" means an alkynyi-alkyl- group in which the alkynyl and alkyl are as previously described. Preferred alkynylalkyls contain a lower alkynyi and a lower alkyl group. The bond to the parent moiety is through the alkyf. Non-limiting examples of suitable alkynylalkyl groups include propargyimethyl. "Heteroaralkyi" means a heteroaryl-alkyl- group in which the heteroaryl and alkyl are as previously described. Preferred heteroaralkyls contain a lower alky! group. Non-limiting examples of suitable araikyl groups include pyridyl methyl, and quinolin-3- ylmethyl. The bond to the parent moiety is through the alkyi.
"Spiro ring systems" have two or more rings linked by one common atom. Preferred spiro ring systems include spiroheteroaryl, spiroheterocyclenyl, spiroheterocyclyl, spirocycloaikyl, spirocyclenyl, and spiroaryl. The spiro ring systems can be optionally substituted by one or more ring system substituents, wherein "ring system substituent" is as defined above. Non-limiting examples of suitable spiro ring
systems include
Figure imgf000155_0003
spiro[4.5jdecane,
Figure imgf000156_0001
8-azaspiro[4.5]dec-2-ene, and '
Figure imgf000156_0002
spiro[4.4]nona-2t7-diene.
Ηydroxyalkyl" means a HO-aikyl- group in which alkyl is as previously defined. Preferred hydroxyalkyls contain lower alkyl. Non-limiting examples of suitable hydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl.
"Acyl" means an H-C(O)-, alkyl-C(O)- or cycloalkyl-C(O)-, group in which the various groups are as previously described. The bond to the parent moiety is through the carbonyl. Preferred acyls contain a lower alkyl. Non-limiting examples of suitable acyl groups include formyl, acetyl and propanoyl. "Aroyl" means an aryl-C(O)- group in which the aryl group is as previously described. The bond to the parent moiety is through the carbonyi. Non-limiting examples of suitable groups include benzoyl and 1- naphthoyl.
"Alkoxy" means an alkyl-O- group in which the alky! group is as previously described. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond to the parent moiety is through the ether oxygen.
"Aryloxy" means an aryl-O- group in which the aryl group is as previously described. Non-ϋmiting examples of suitable aryloxy groups include phenoxy and naphthoxy. The bond to the parent moiety is through the ether oxygen. "Aralkyloxy" means an aralkyl-O- group in which the aralkyi group is as previously described. Non-limiting examples of suitable aralkyloxy groups include benzyloxy and 1- or 2-naphthalenemethoxy. The bond to the parent moiety is through the ether oxygen.
"Alkylthio" means an alkyl-S- group in which the aikyl group is as previously described. Non-limiting examples of suitable alkylthio groups include methylthio and ethylthio. The bond to the parent moiety is through the sulfur.
"Arylthio" means an aryl-S- group in which the aryl group is as previously described. Non-limiting examples of suitable arylthio groups include phenyithio and naphthylthio. The bond to the parent moiety is through the sulfur. "Aralkylthio" means an aralkyl-S- group in which the aralkyl group is as previously described. Non-limiting example of a suitable aralkylthio group is benzylthio. The bond to the parent moiety is through the sulfur.
"Alkoxycarbonyl" means an alkyi-O-CO- group. Non-limiting examples of suitable alkoxycarbonyl groups include methoxycarbonyl and ethoxycarbonyl. The bond to the parent moiety is through the carbonyl.
"Aryloxycarbonyl" means an aryl-O-C(O)- group. Non-limiting examples of suitable aryloxycarbonyi groups include phenoxycarbonyl and naphthoxycarbonyi. The bond to the parent moiety is through the carbonyl. "Aralkoxycarbonyl" means an aralkyl-O-C(O)- group. Non-limiting example of a suitable aralkoxycarbonyl group is benzyloxycarbonyl. The bond to the parent moiety is through the carbonyl.
"Alkylsulfonyi" means an alkyi-S(O2)- group. Preferred groups are those in which the alkyl group is lower alkyl. The bond to the parent moiety is through the sulfonyl.
"Arylsulfonyl" means an aryl-S(O2)- group. The bond to the parent moiety is through the sulfonyl.
The term "substituted" means that one or more hydrogen on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency under the existing circumstances is not exceeded, and that the substitution results in a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds. By "stable compound' or "stable structure" is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
The term "optionally substituted" means optional substitution with the specified groups, radicals or moieties.
The term "purified", "in purified form" or "in isolated and purified form" for a compound refers to the physical state of said compound after being isolated from a synthetic process (e.g. from a reaction mixture), or natural source or combination thereof. Thus, the term "purified", "in purified form" or "in isolated and purified form" for a compound refers to the physical state of said compound after being obtained from a purification process or processes described herein or wefl known to the skilled artisan (e.g., chromatography, recrystallization and the like) , in sufficient purity to be characterizable by standard analytical techniques described herein or well known to the skilled artisan. It should aiso be noted that any carbon as well as heteroatom with unsatisfied valences in the text, schemes, examples and Tables herein is assumed to have the sufficient number of hydrogen atom(s) to satisfy the valences.
When a functional group in a compound is termed "protected", this means that the group is in modified form to preclude undesired side reactions at the protected site when the compound is subjected to a reaction. Suitable protecting groups will be recognized by those with ordinary skill in the art as well as by reference to standard textbooks such as, for example, T. W. Greene et a/, Protective Groups in organic Synthesis (1991), Wiley, New York.
When any variable (e.g., aryl, heterocycle, R2, etc.) occurs more than one time in any constituent or in The present invention, its definition on each occurrence is independent of its definition at every other occurrence.
As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
Prodrugs and solvates of the compounds of the invention are also contemplated herein. A discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press. The term "prodrug" means a compound (e.g., a drug precursor) that is transformed in vivo Xo yieid a compound of the present invention or a pharmaceutically acceptable salt, hydrate or solvate of the compound. The transformation may occur by various mechanisms (e.g., by metabolic or chemical processes), such as, for example, through hydrolysis in blood. A discussion of the use of prodrugs is provided by T. Higuchi and W. Stella, "Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutica! Association and Pergamon Press, 1987.
For example, if a compound of the present invention or a pharmaceutically acceptable salt, hydrate or solvate of the compound contains a carboxyiic acid functional group, a prodrug can comprise an ester formed by the replacement of the hydrogen atom of the acid group with a group such as, for example, (C1-Cβjalkyl, (C2- Ci2)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1- methyl-1 -(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms, 1 -(N-(alkoxycarbonyl)amino)ethy! having from 4 to 10 carbon atoms, 3-phthalidyl, 4- crotonolactonyl, gamma-butyrolacton-4-yl, di-N, N-(Ci -C2)alkylamino(C2-C3)alkyl (such as β-dimethylaminoethyl), carbamoyl-fC1-C^alkyl, N,N-di (C1-C2)alkyicarbamoyi-(C1- C2)alkyl and piperidino-, pyrrolidino- or morpholino{C2-C3)alkyl, and the like.
Similarly, if a compound of the present invention contains an alcohol functional group, a prodrug can be formed by the replacement of the hydrogen atom of the alcohol group with a group such as, for example, (C1-C6)alkanoyloxymethyl, 1 -((Ci- C6)alkanoyloxy)ethyl, 1 -methyl-1 -((C1-C6)alkanoyloxy)ethyl, (C1- C6)alkoxycarbonyloxymethyl, N-(C1-C6)alkoxycarbonylaminomethyl, succinoyl, (Cr C6)alkanoy!, α-amino(C1-C4)alkanyl, aryiacyl and α-aminoacyl, or α-aminoacyl-α- aminoacyl, where each α-aminoacyl group is independently selected from the naturally occurring L-amino acids, P(O)(OH)2, -P(O)(O(Ci -C6)alkyl)2 or glycosyl (the radical resulting from the removal of a hydroxyl group of the hemiacetal form of a carbohydrate), and the like.
If a compound of the present invention incorporates an amine functional group, a prodrug can be formed by the replacement of a hydrogen atom in the amine group with a group such as, for example, R-carbonyi, RO-carbonyl, NRR'-carbonyl where R and R' are each independently (Ci~Cio)alkyt, (C3-C7) cycloalkyl, benzyl, or R-carbonyl is a natural α-aminoacyl or natural α-aminoacyl, -C(OH)C(O)OY1 wherein Y1 is H, (C1-Ce)alkyl or benzyl, -C(OY2JY3 wherein Y2 is (C1-C4) alky] and Y3 is (C1-C6)alkyl, carboxy (C1-C6)alky!, amino(C1-C4)alkyl or mono-N — or di~N,N-(C1-C6)alky!aminoalkyl, -C(Y4) Y5 wherein Y4 is H or methyl and Y5 is mono-N— or di-N,N-(C1-C6)alkylamino morpholino, piperidin-1-yl or pyrroiidin-1 -yl, and the like.
One or more compounds of the invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable soivents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms. "Solvate" means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. "Solvate" encompasses both solution-phase and isolatable solvates. Non-limiting examples of suitable solvates include ethanoiates, methanolates, and the like. "Hydrate" is a solvate wherein the solvent molecule is H2O.
One or more compounds of the invention may optionally be converted to a solvate. Preparation of solvates is generally known. Thus, for example, M. Caira et al, J. Pharmaceutical Sci., 93(3), 601-611 (2004) describes the preparation of the solvates of the antifungal fluconazole in ethyl acetate as well as from water. Similar preparations of solvates, hemisolvate, hydrates and the like are described by E. C. van Tonder et al, AAPS PharmSciTech., 5£1), article 12 (2004); and A. L. Bingham et al, Chem. Commυn., 603-604 (2001). A typical, non-limiting, process involves dissolving the inventive compound in desired amounts of the desired solvent (organic or water or mixtures thereof) at a higher than ambient temperature, and cooling the solution at a rate sufficient to form crystals which are then isolated by standard methods. Analytical techniques such as, for example I. R. spectroscopy, show the presence of the solvent (or water) in the crystals as a solvate (or hydrate).
"Effective amount" or "therapeutically effective amount" is meant to describe an amount of compound or a composition of the present invention effective in inhibiting the above-noted diseases and thus producing the desired therapeutic, ameliorative, inhibitory or preventative effect. The compounds of the present invention can form salts which are also within the scope of this invention. Reference to a compound of the present invention herein is understood to include reference to salts thereof, unless otherwise indicated. The term "saltfs)", as employed herein, denotes acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases. In addition, when a compound of the present invention contains both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions ("inner salts") may be formed and are included within the term "salt(s)" as used herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful. Salts of the compounds of the The present invention may be formed, for example, by reacting a compound of the present invention with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.
Exemplary acid addition salts include acetates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromides, hydroiodides, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartarates, thiocyanates, toluenesulfonates (also known as tosylates,) and the like. Additionally, acids which are generally considered suitable for the formation of pharmaceutically useful salts from basic pharmaceutical compounds are discussed, for example, by P. Stahl et al, Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiiey-VCH; S. Berge et al, Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201 -217; Anderson et al, The Practice of Medicinal Chemistry (1996), Academic Press, New York; and in The Orange Book (Food & Drug Administration, Washington, D. C. on their website). These disclosures are incorporated herein by reference thereto.
Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamines, t-butyl amines, and salts with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups may be quartemized with agents such as lower alkyl hatides (e.g. methyl, ethyl, and butyl chlorides, bromides and iodides), dialky! sulfates (e.g. dimethyl, diethyl, and dibutyl sulfates), long chain halides (e.g. decyl, iauryl, and stearyl chlorides, bromides and iodides), aralkyl haiides (e.g. benzyl and phenethyl bromides), and others.
All such acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention.
Pharmaceutically acceptable esters of the present compounds include the following groups: (1 ) carboxylic acid esters obtained by esterification of the hydroxy groups, in which the non-carbonyl moiety of the carboxyiic acid portion of the ester grouping is selected from straight or branched chain alkyl (for example, acetyl, n- propyl, t-butyl, or n-butyl), alkoxyalkyl (for example, methoxymethyl), araikyl (for example, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (for example, phenyl optionally substituted with, for example, halogen, d^alkyl, or Ci.4aikoxy or amino); (2) sulfonate esters, such as alkyl- or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters (for example, L-valyl or L-isoieucyl); (4) phosphonate esters and (5) mono-, di- or triphosphate esters. The phosphate esters may be further esterified by, for example, a C1-2o alcohol or reactive derivative thereof, or by a 2,3-di (C6-24)acyl glycerol.
Compounds of the present invention, and salts, solvates, esters and prodrugs thereof, may exist in their tautomeric form (for example, as an amide or imino ether). All such tautomeric forms are contemplated herein as part of the present invention.
The compounds of the present invention may contain asymmetric or chiral centers, and, therefore, exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the present invention as weil as mixtures thereof, including racemic mixtures, form part of the present invention. In addition, the present invention embraces all geometric and positional isomers. For example, if a compound of the present invention incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the invention. Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods weil known to those skilled in the art, such as, for example, by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydroiyzing) the individual diastereomers to the corresponding pure enantiomers. Also, some of the compounds of the present invention may be atropisomers (e.g., substituted biaryls) and are considered as part of this invention. Enantiomers can also be separated by use of chiral HPLC column.
It is also possible that the compounds of the present invention may exist in different tautomeric forms, and all such forms are embraced within the scope of the invention. Also, for example, all keto-enol and imine-enamine forms of the compounds are included in the invention.
All stereoisomers (for example, geometric isomers, optical isomers and the like) of the present compounds {including those of the salts, solvates, esters and prodrugs of the compounds as well as the salts, solvates and esters of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated within the scope of this invention, as are positional isomers (such as, for example, 4-pyridyl and 3-pyridyl). (For example, if a compound of the present invention incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the invention. Also, for example, all keto-enol and imine-enamine forms of the compounds are included in the invention.) Individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. The chiral centers of the present invention can have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of the terms "salt", "solvate", "ester", "prodrug" and the like, is intended to equally apply to the salt, solvate, ester and prodrug of enantiomers, stereoisomers, rotamers, tautomers, positional isomers, racemates or prodrugs of the inventive compounds.
The present invention also embraces isotopically-labelled compounds of the present invention which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine and iodine, such as 2H, 3H, 11C, 13C, 14C, 15N, 18O, 17O, 31P, 32P, 35S1 18F, 36CI and 123I, respectively.
Certain isotopically-labelled compounds of Formula (!) (e.g., those labeled with 3H and 14C) are useful in compound and/or substrate tissue distribution assays. Tritiated {i.e., 3H) and carbon-14 (i.e., 14C) isotopes are particularly preferred for their ease of preparation and detectabiiity. Certain isotopically-labelled compounds of Formula (I) can be useful for medical imaging purposes. E.g., those labeled with positron-emitting isotopes like 11C or 18F can be useful for application in Positron Emission Tomography (PET) and those labeled with gamma ray emitting isotopes like 123I can be useful for application in Single photon emission computed tomography (SPECT). Further, substitution with heavier isotopes such as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances. Further, substitution with heavier isotopes such as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances. Additionally, isotopic substitution at a site where epimerization occurs may slow or reduce the eptmerization process and thereby retain the more active or efficacious form of the compound for a longer period of time, isotopically labeled compounds of Formula (I), in particular those containing isotopes with longer half lives (T1/2 >1 day), can generally be prepared by following procedures analogous to those disclosed in the Schemes and/or in the Examples herein below, by substituting an appropriate isotopically labeled reagent for a non-isotopically labeled reagent.
Polymorphic forms of the compounds of the present invention, and of the salts, solvates, esters and prodrugs of the compounds of the present invention, are intended to be included in the present invention. The compounds according to the invention have pharmacological properties; in particular, the compounds of the present invention can be inhibitors, regulators or modulators of mTOR protein kinases.
The compounds of the present invention can be inhibitors of protein kinases such as, for example, the inhibitors of the mTOR. Preferred compounds can exhibit IC50 values of less than about 5μm, preferably about 0.001 to about 1.0 μm, and more preferably about 0.001 to about 0.1 μm. The assay methods are described in the Examples set forth below.
The compounds of the present invention can be useful in the therapy of proliferative diseases such as cancer, autoimmune diseases, viral diseases, fungal diseases, neurological/neurodegenerative disorders, arthritis, inflammation, antiproliferative (e.g., ocular retinopathy), neuronal, alopecia and cardiovascular disease. Many of these diseases and disorders are iisted in U.S. 6,413,974, incorporated by reference herein. More specif icaliy, the compounds of the present invention can be useful in the treatment of a variety of cancers, including (but not limited to) the following: tumor of the bladder, breast (including BRCA-mutated breast cancer, colorectal, colon, kidney, liver, lung, small cell lung cancer, non-smail cell lung cancer, head and neck, esophagus, bladder, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, including squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma and Burkett's lymphoma; chronic lymphocytic leukemia ("CLL"), acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocytic leukemia; fibrosarcoma, rhabdomyosarcoma; head and neck, mantle cell lymphoma, myeloma; astrocytoma, neuroblastoma, glioma, glioblastoma, malignant glial tumors, astrocytoma, hepatocellular carcinoma, gastrointestinal stromal tumors ("GIST") and schwannomas; melanoma, multiple myeloma, seminoma, teratocarcinoma, osteosarcoma, xenoderma pigmentosum, keratoctanthoma, thyroid follicular cancer, endometrial cancer, gastrointestinal tract cancer and Kaposi's sarcoma.
Due to the key roie of kinases in the reguiation of cellular proliferation in general, inhibitors could act as reversible cytostatic agents which may be useful in the treatment of any disease process which features abnormal cellular proliferation, e.g., benign prostate hyperplasia, familial adenomatosis polyposis, neuro-fibromatosis, atherosclerosis, pulmonary fibrosis, arthritis, psoriasis, glomerulonephritis, restenosis following angioplasty or vascular surgery, hypertrophic scar formation, inflammatory bowel disease, transplantation rejection, endotoxic shock, and fungal infections. Compounds of the present invention may induce or inhibit apoptosis. The apoptotic response is aberrant in a variety of human diseases. Compounds of the present invention, as modulators of apoptosis, will be useful in the treatment of cancer (including but not limited to those types mentioned hereinabove), virai infections (including but not limited to herpevirus, poxvirus, Epstein- Barr virus, Sindbis virus and adenovirus), prevention of AIDS development in HIV-infected individuals, autoimmune diseases (including but not limited to systemic lupus, erythematosus, autoimmune mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes mellitus), neurodegenerative disorders (including but not limited to Alzheimer's disease, AIDS-reiated dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy and cerebellar degeneration), myelodysplasia syndromes, aplastic anemia, ischemic injury associated with myocardial infarctions, stroke and reperfusion injury, arrhythmia, atherosclerosis, toxin-induced or alcohol related liver diseases, hematological diseases (including but not limited to chronic anemia and aplastic anemia), degenerative diseases of the musculoskeletal system (including but not limited to osteoporosis and arthritis) aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis, kidney diseases and cancer pain.
Compounds of the present invention, as inhibitors of kinases, can modulate the level of cellular RNA and DNA synthesis. These agents would therefore be useful in the treatment of viral infections (including but not limited to HIV, human papilloma virus, herpesvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus). Compounds of the present invention may also be useful in the chemoprevention of cancer. Chemoprevention is defined as inhibiting the development of invasive cancer by either blocking the initiating mutagenic event or by blocking the progression of pre-malignant cells that have already suffered an insult or inhibiting tumor relapse.
Compounds of the present invention may also be useful in inhibiting tumor angiogenesis and metastasis.
Another aspect of this invention is a method of treating a mammal {e.g., human) having a disease or condition associated with mTOR kinases by administering a therapeutically effective amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound to the mammal.
A preferred dosage is about 0.001 to 1000 mg/kg of body weight/day of the compound of the present invention. An especially preferred dosage is about 0.01 to 25 mg/kg of body weight/day of a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound. The compounds of this invention may also be useful in combination (administered together or sequentially) with one or more of anti-cancer treatments such as radiation therapy, and/or one or more anti-cancer agents different from the compound of the present invention. The compounds of the present invention can be present in the same dosage unit as the anti-cancer agent or in separate dosage units.
Another aspect of the present invention is a method of treating one or more diseases associated with a mTOR protein kinase, comprising administering to a mammal in need of such treatment: an amount of a first compound, which is a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof; and an amount of at least one second compound, the second compound being an anti-cancer agent different from the compound of the present invention, wherein the amounts of the first compound and the second compound result in a therapeutic effect. Non-limiting examples of suitable anti-cancer agent is selected from the group consisting of a Cytostatic agent, Cisplatin, Deforolimus (described in PCT publication No. 2003/064383), Doxorubicin, liposomal doxorubicin {e.g., Caelyx®, Myocet®, Doxil®), Taxotere, Taxol, Etoposide, frinotecan, Camptostar, Topotecan, Paclitaxel, Docetaxel, Epothilones, Tamoxifen, 5-Fluorouracil, Methoxtrexate, Temozolomide, cyclophosphamide, SCH 66336, R115777®, L778.123®, BMS 214662®, iressa®, Tarceva®, Antibodies to EGFR, antibodies to IGFR (including, for example, those published in US 2005/0136063 published June 23, 2005), ESK inhibitors, KSP inhibitors (such as, for example, those published in WO 2006/098962 and WO 2006/098961 ; ispinesib, SB-743921 from Cytokinetics), Centrosome associated protein E ("CENP-E") inhibitors (e.g., GSK-923295), Gleevec®, Intron, Ara-C, Adriamycin, Cytoxan, Gemcitabine, Uracil mustard, Chlormethine, ifosfamide, Melphalan, Chlorambucil, Pipobroman, Triethylenemelamine,
Triethylenethiophosphoramine, Busulfan, Carmustine, Lomustine, Streptozocin, Dacarbazine, Floxuridine, Cytarabine, 6-Mercaptopurine, 6-Thioguanine, Fludarabine phosphate, Oxaliplatin, Leucovirin, ELOXATIN™, Vinblastine, Vincristine, Vindesine, Bleomycin, Dactinomycin, Daunorubicin, Doxorubicin, Epirubicin, Idarubicin, Mithramycin, Deoxycoformycin, Mitomycin-C, L-Asparaginase, Teniposide 17oc- Ethinylestradiol, Diethylstilbestroi, Testosterone, Prednisone, Fluoxymesterone, Dromostanolone propionate, Testolactone, Megestrolacetate, Methylprednisoione, Methyltestosterone, Prednisolone, Triamcinolone, Chlorotrianisene, Hydroxyprogesterone, Aminoglutethimide, Estramustine, Medroxyprogesteroneacetate, Leuprolide, Flutamide, Toremifene, Goserelin, Cisplatin, Carboplatin, Hydroxyurea, Amsacrine, Procarbazine, Mitotane, Mitoxantrone, Levamisole, IMavelbene, Anastrazole, Letrazole, Capecitabine, Reioxafine, Droloxafine, Hexamethylmelamine, Avastin, herceptin, Bexxar, bortezomib ("Velcade"), Zevalin, Trisenox, Xeloda, Vinorelbine, Porfimer, Erbitux, Liposomal, Thiotepa, Altretamine, Melphalan, Trastuzumab, Lerozole, Fulvestrant, Exemestane, Fulvestrant, Ifosfomide, Rituximab, C225®, Satriplatin, mylotarg, Avastin, Rituxan, Panitubimab, Sutent, Sorafinib, Sprycel (dastinib), Nilotinib, Tykerb (Lapatinib) and Campath.
If formulated as a fixed dose, such combination products employ the compounds of this invention within the dosage range described herein and the other pharmaceutically active agent or treatment within its dosage range. For example, the CDC2 inhibitor olomucine has been found to act synergistically with known cytotoxic agents in inducing apoptosis (J. CeII ScL, (1995) 108, 2897. Compounds of the present invention may also be administered sequentially with known anticancer or cytotoxic agents when a combination formulation is inappropriate. The invention is not limited in the sequence of administration; compounds of the present invention may be administered either prior to or after administration of the known anticancer or cytotoxic agent. For example, the cytotoxic activity of the cyclin-dependent kinase inhibitor flavopiridol is affected by the sequence of administration with anticancer agents. Cancer Research, (1997) 57, 3375. Such techniques are within the skills of persons skiiled in the art as well as attending physicians. Accordingly, in an aspect, this invention includes combinations comprising an amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof, and an amount of one or more anticancer treatments and anti-cancer agents listed above wherein the amounts of the compounds/ treatments result in desired therapeutic effect. Another aspect of the present invention is a method of inhibiting one or more mTOR protein kinases in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of at least one compound of the present invention or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
Another aspect of the present invention is a method of treating, or slowing the progression of, a disease associated with one or more mTOR protein kinases in a patient in need thereof, comprising administering a therapeutically effective amount of at least one compound of the present invention or a pharmaceuticaily acceptable salt, solvate, ester or prodrug thereof.
Yet another aspect of the present invention is a method of treating one or more diseases associated with mTOR protein kinases, comprising administering to a mammai in need of such treatment an amount of a first compound, which is a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof; and an amount of at least one second compound, the second compound being an anti-cancer agent, wherein the amounts of the first compound and the second compound result in a therapeutic effect.
Another aspect of the present invention is a method of treating, or slowing the progression of, a disease associated with one or more mTOR protein kinases in a patient in need thereof, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising in combination at least one pharmaceutically acceptable carrier and at least one compound according to The present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
In the above methods, the mTOR protein kinases to be inhibited can exist in two complexes, mTORC1 and mTORC2.
ERK inhibitors (i.e., ERK1 inhibitors and/or ERK2 inhibitors) include but are not limited to the following compounds: a) Formula H
Figure imgf000170_0001
or the pharmaceutically acceptable salts, esters and solvates thereof, wherein:
Y1 , Y2, and Y3 are each independently selected from the group consisting of: -CH=, -N= and -CR9=; z is 1 to 3; Q is a substituent selected from the group consisting of:
(2.8)
Figure imgf000170_0002
(2.13) ) (2.16)
Figure imgf000171_0001
Each Q1 represents a ring independentiy selected from the group consisting of: cycloalkyl, substituted cycloalkyi, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted ary!, heteroaryl, and substituted heteroaryl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: halo and the R10 moieties; provided that when Q1 is ary!, heteroaryl, substituted aryl or substituted heteroaryl then the carbon atoms at the ring junction are not substituted; Q2 represents a ring selected from the group consisting of: cycloalkyl, substituted cycloaikyl, heterocycloaikyl, and substituted heterocycloalkyl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R10 moieties;
Z1 represents -(C(R24)2)W- wherein each R24 is independently selected from the group consisting of: H, alkyl and F, and wherein w is 1 , 2 or 3;
Z2 is selected from the group consisting of: -N(R44)-, -O- and -C(R46J2-; m is 1 to 6; n is 1 to 6; p is 0 to 6; t is O1 1 , or 2;
R1 is selected from the group consisting of: (I) -CN,
(2) -NO2,
(3) -OR10, (4) -SR10,
(5) -N(R10J2,
(6) R10,
(7) -C(O)R10,
(8) -(C(R30)2)n-NR32-C(O)-R10, (9) -(C(R30)2)n-NR32-S(O)rR10,
(10) -(C(R30)2)n-NR32-C(O)-N(R32)-R10,
(11 )
Figure imgf000172_0001
(12) -CF3, (13) -C(O)OR10,
(14) -(C(R30)2)nR13 (β.g., -(CH2)nR13),
(15) alkenyl, (16) -NR32-C(O)-R14, (17)
Figure imgf000173_0003
wherein each R10 is independently selected, (18)
Figure imgf000173_0004
wherein each R10 is independently selected, (19)
Figure imgf000173_0001
(20) -C(O)-N R32-(C(R30)2)p-OR10,
(21 ) -C(O)N(R10)2 wherein each R10 is independently selected, (22) -C(O)-N R32-C(R18)3,
(23) -C(O)-NR32-(C(R30)2)n-C(O)-N(R10)2,
(24) heterocycloalkenyl,
(25)
Figure imgf000173_0002
(26) arylalkenyl-;
R2 is selected from the group consisting of: (D H1
(2) -CN,
(3) halo, (4) alkyl, (5) substituted alkyl wherein said substituted aikyl is substituted with 1 to 3 substitutents selected from the group consisting of: (a) -OH, (b) -O-alkyl (e.g., -0-(Cr C3a!ky!), (c) -O-alkyl substituted with 1 to 3 F atoms, and (d) -N(R40)2 wherein each R40 is independently selected from the group consisting of: (i) H, (ii) C1-C3 alkyl, (iii) - CF3, and (e) halo,
(6) alkynyl,
(7) alkenyl, <8) -(CHΞ)mR1\ (9) -N(R26)2> (1 O) -OR23,
(11 ) -N(R26JC(O)R42, (12) cycloaikyl, (13) cycloaikylalkyl,
Figure imgf000174_0001
(15) -O-(substituted alkyl) wherein said substituted alkyi is substituted with
1 to 3 F atoms,
(16) -S(0)ralkyl, <17) -C(O)-alkyl,
(18)
N— O— H £ Il ξ — C— alkyl >
(19)
N— O— alkyl I — C— alkyl
wherein each alkyi is independently selected,
(20)
O
H I! N—N— C— aikyl
< — C— alkyi > which each alkyl is independently selected, (21 )
Figure imgf000175_0001
wherein each alkyl is independently selected,
(22) -N(R48)-C(O)-R48 wherein each R48 is independently selected from the group consisting of: H and alkyl, and
(23) -C(O)-alkyl, such as, for example, -C(O)-(C1-C6 alky!), such as, for example, -C(O)CH3; each R3, R4, R5, R6 and R7 is independently selected from the group consisting of: (I) H1
(2) alkenyl,
(3) substituted alkenyl,
(4) alkyl,
(5) substituted aikyl, (6) cycloalkyl,
(7) substituted cycioalkyl,
(8) cycloaikylalkyl-,
(9) substituted cycloaikylalkyl-,
(10) heterocycloalkyl, (11 ) substituted heterocycloalkyl,
(12) heterocycloalkylalkyl-,
(13) substituted heterocycloalkylalkyl-,
(14) -C(O)R10,
(15) arylheteroaryl-, (16) substituted arylheteroaryl-,
(17) heteroarylaryl-,
(18) substituted heteroarylaryl-, (19) aryl,
(20) substituted aryl, (21) heteroaryl,
(22) substituted heteroaryi,
(23) heteroarylheteroaryi-,
(24) substituted heteroarylheteroaryi-, (25) arylaminoheteroaryl-,
(26) substituted arylaminoheteroaryl-,
(27) aryialkynyl-,
(28) substituted arylalkynyi-,
(29) heteroarylalkynyl-, (30) substituted heteroarylaikynyl-, and
(31) benzoheteroaryl, wherein said R3 T R4, R5, R6 and R7 substituted groups (7), (9), (11), (13), (16), (18), (20), (22), (24), (26), (28) and (30) are substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NHR20, -N(R20J2 wherein each R20 is independently selected, alkyl, alkenyl, halo, -C(O)-NH-R26, -C(O)OR28, -C(O)R28, and -OR20, and wherein said R3, R4, R5, R6 and R7 substituted groups (3) and (5) are substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, halo (e.g., F, Cl and Br, and in another example F), -C(O)-NH-R28 (e.g., -C(O)-NH-CH3), -C(O)OR28 (e.g., -C(O)OC2H5), and -C(O)R28 (e.g., -C(O)CH3); R5A is selected from the group consisting of: haio, -OH, alkyt, and -O-alkyl; R8 is selected from the group consisting of: H, -OH, -N(R10J2, -NR1°C(O)R12, and alkyl; each R9 is independently selected from the group consisting of:halogen, -CN, -NO2, -OR10, -SR10, -N(R10J2, and R10; each R10 is independently seiected from the group consisting of: H, alkyi, aryl, arylalkyl, heteroaryi, heteroaryl alkyl, cycioalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyi, alkyl heteroaryl-, alkylaryl-, substituted alkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycioalkyl, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycloalkylalkyi, substituted alkyiheteroaryh substituted alkylaryl-, heterocycioalkenyl, and substituted heterocycloalkenyl, and wherein: said R10 substituted alkyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NHR20, -NO2, -CN, -OR26, halo, -C(O)-NH-R26, -C(O)OR26, and -C(O)R26, and said R10 substituted aryl, substituted arylalkyi, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloalkyi, substituted cycloaikylalkyl, substituted heterocycloalkyl, substituted heterocycloalkylaikyi, substituted aikylheteroaryl- and substituted alkyiaryl- are substituted with 1 to 3 substituents independently selected from the group consisting of: (1) -NH2, (2) -NO2, (3) -CN, (4) -OH, (5) -OR20, (6) -OCF3, (7) alkyl substituted with 1 to 3 independently selected halo atoms, (8) -C(O)R33, (9) alkyl, (10) alkenyl, (11 ) halo, (12) -C(O)-NH-R26, (13) -C(O)OR38, (14) -C(O)-NR32-(C(R30)2)n-N(R38)2, (15) -S(O)1R38, (16) -C(O)-NR32-R38, (17) -NR32-C(O)-R38,
Figure imgf000177_0001
(19) -NHR20, (20) cycloalkyi, (21 ) -O-alkyl-O-R20, (22) hydroxyalkyl, (23) -N(R20J2 wherein each R20 is independently selected, (24) -alkyl-OR20, (25) -0-alkyi-OH, (26) -NH(hydroxyalkyl), and (27) oxazolidinone;
R11 is selected from the group consisting of: F, -OH, -CN, -OR10, -NHNR1R10, -SR10 and heteroaryl;
R12 is selected from the group consisting of: alkyl, aryl, heteroaryl, cycloalkyi, eye! oa Iky I alkyl, heterocycloalkyl and heterocycloalkylaikyi;
R14 is selected from the group consisting of: alkyt, aryl, heteroaryl, cycloalkyi, cycloaikylalkyl-, heterocycloalkyl, alkylheterocycloalkyl, heterocycioalkylalkyl-, aikylheteroaryi- and aikyiaryl-;
R15 is selected from the group consisting of: H, -OH, alkyl, aryl, heteroaryl, cycloalkyl, cycloaikylalkyl-, heterocycloalkyl and heterocycloalkylaikyi-, alkylheteroaryl- and alkylary!-;
R20 represents alkyl;
R23 is selected from the group consisting of: H, alkyl, aryl, cycloaikyl, and cycloalkylalkyi-; each R26 is independently selected from the group consisting of: H and alkyl;
R28 is alkyi; each R30 is independently selected from the group consisting of: H, alkyl, and F; each R32 is independently selected from the group consisting of: H and alkyl, and wherein each R32 is generally H; each R35 is independently selected from the group consisting of: H and Ci to C6 aikyl;
R36 is selected from the group consisting of: H, alkyl, and -O-alkyl; each R38 is independently selected from the group consisting of: H, alkyl, aryi, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalky! alkyl, heterocycloalkyl, heterocycloalkylalkyl, alkylheteroaryl-, alkylaryl-, substituted alkyl, substituted aryl, substituted arylaikyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloalkyl, substituted cycloalkylalkyl, substituted heterocycloalkyi, substituted heterocycloalkylalkyi, substituted alkylheteroaryl- and substituted alkylaryl-, and wherein: said R38 substituted alkyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NO2, -CN, -OR26, halo, -C(O)-NH-R28, -C(O)OR28, and -C(O)R28, and said R3δ substituted aryl, substituted aryialkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloaikyl, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycloalkylalkyl, substituted alkylheteroaryl- and substituted alkylaryl- are substituted with 1 to 3 substituents independently selected from the group consisting of: (1) -NH2, (2) -NO2, (3) -CN, (4) -OH, (5) -OR20, (6) -OCF3, (7) -CF3, (8) -C(O)R26, (9) aikyl, (10) alkenyl, (11) halo, (12) -C(O)-NH-R26, (13) -C(O)OR26, (14) -C(O)-NR32-{C(R30)2)π-N(R26)2, (15) -S(O)1R26, (16) -C(O)N(R32J(R26), (17) -NR32C(O)R26,
Figure imgf000178_0001
(19) -NHR20;
R42 is selected from the group consisting of: alkyl, aryl, heteroaryl, and cycloalkyl; R44 is selected from the group consisting of: H, alkyl, cycloalkyl, and cycloaikylalkyl; Each R46 is independently selected from the group consisting of: H, alkyi, cycloalkyl, and cycloalkylalkyl; and provided that:
(1) Q1 is a substituted ring wherein at least one substitutent is halo; and/or (2) R2 is substituted alkyl wherein said substituent is -OCH3, or R2 is selected from the group consisting of: -CH2OH and -CH2OCH3; and/or
(3) at least one of R3, R4, R5, R6, and R7 is:
(i) selected from the group consisting of: oxadiazolylphenyl-, pyridazinylphenyl-, pyrimidinylpyrazinyl-, substituted oxadiazolylphenyl-, substituted pyridazinylphenyl-, substituted pyrimidinylpyrazinyi-, and benzoheteroaryl; or (ii) selected from the group consisting of:
(a) substituted cycloalkyl,
(b) substituted cycloalkyialkyl-,
(c) substituted heterocycloaikyl, <d) substituted heterocycloalkylalkyl-,
(e) substituted arylheteroaryl-,
(f) substituted heteroarylaryl-,
(g) substituted aryl,
(h) substituted heteroaryl, (i) substituted heteroarylheteroaryl-,
G) substituted arylaminoheteroaryl-, (k) substituted arylalkynyl-, and (!) substituted heteroarylalkynyl-, and wherein at least one substitutent on at least one at least one of said (ii)(a) to (U)(I) R3, R4, R5, R6, or R7 group is selected from the group consisting of: -NHR20, -N(R20J2 (wherein each Rao is independently selected), and -OR20; and/or
(4) R5A is alkyl; and/or
(5) R10 is a substituted aryl, and at least one substitutent is selected from the group consisting of: (a) -S(O)tR38 wherein R38 is isopropyl,
(b) -O-alkyl-O-R20,
(c) hydroxyalkyl,
Figure imgf000180_0001
(e) -aikyl-OR20,
(f) -O-alkyl-OH,
(g) -NH(hydroxyalkyl), and (h) oxazolidinone; and/or
(6) R20 is isopropyl;
Formula Il is described in PCT publication No. 2007/070398, herein incorporated by reference; b) Formula III
Figure imgf000180_0002
II! or the pharmaceutically acceptable salts, esters or solvates thereof, wherein: z is 1 to 3; Q is a substituent selected from the group consisting of:
Figure imgf000180_0003
( ) ( (2.7) (2.8) (2.13) ( 15) (2.16)
(2.20) (2.22)
Figure imgf000181_0001
Each Q1 represents a ring independently selected from the group consisting of: cycioalkyl, substituted cycioaikyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, and substituted heteroaryl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: haio and the R10 moieties; provided that when Q1 is ary!, heteroaryl, substituted aryl or substituted heteroaryl then the carbon atoms at the ring junction are not substituted;
Q2 represents a ring selected from the group consisting of: cycloalkyl, substituted cycloalkyi, heterocycloalkyl, and substituted heterocycloalkyl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R10 moieties;
Z1 represents -{C(R24)2)W- wherein each R24 is independently selected from the group consisting of: H, alkyl and F, and wherein w is 1 , 2 or 3; Z2 is selected from the group consisting of: -N(R44)-, -O- and -C(R46)2-; m is 1 to 6; n is 1 to 6; p is 0 to 6; t is 0, 1 , or 2; R1 is selected from the group consisting of:
O) -CN,
(2) -NO2,
(3) -OR10,
(4) -SR10, (5) -N(R1V
(6) R10,
(7) -C(O)R10,
(8) -(C(R30)2)n-NR32-C(O)-R10, wherein in one example n is 1 , each R30 is H, R32 is H, and R10 is selected from the group consisting of: cycloalkyl and alkyl, (9) -(C(R30)2)n-NR32-S(O)rR10,
(10) -(C(R30)2)n-NR32-C(O)-N(R32}-R10,
(11)
Figure imgf000182_0001
(12) -CF3, (13) -C(O)OR10,
(14) -(C(R30)2)nR13,
(15) alkenyl (e.g., -CH=CHCH3),
(16) -NR32-C(O)-R14, (17)
Figure imgf000183_0004
wherein each R10 is independently selected, (18)
Figure imgf000183_0005
wherein each R10 is independently selected, (19)
Figure imgf000183_0001
(20) -C(O)-NR32-(C(R30)2)p-OR10,
(21 ) -C(O)N(R10)2 wherein each R10 is independently selected, (22) -C(O)-NR32-C(R18)3,
(23) -C(O)-NR32-(C(R30)2)n-C(O)-N(R10)2l (24) heterocycloalkenyl, such as, for example:
Figure imgf000183_0002
wherein r is 1 to 3, (25)
Figure imgf000183_0003
(26) arylalkenyl-, and
(27) halo; R2 is selected from the group consisting of: (I) H1
(2) -CN,
(3) halo, (4) alkyl,
(5) substituted alkyl wherein said substituted alkyl is substituted with 1 to 3 substitutents selected from the group consisting of: (a) -OH, (b) -O-alkyl, (c) -O-alkyl substituted with 1 to 3 F atoms, and (d) -N(R40)2 wherein each R40 is independently selected from the group consisting of: (i) H, (ii) C1-C3 alkyl, (iii) -CF3, and (e) halo, (6) alkynyl,
(7) alkenyl,
(8) -(CH2UR1 \
(9) -N(R26)2) (1O) -OR23, (11 ) -N(R26)C(O)R42,
(12) cycloalkyl, (13) cycloalkylalkyl,
(14)
Figure imgf000184_0001
(15) -O-(substituted alkyl) wherein said substituted alkyl is substituted with 1 to 3 F atoms,
(16) -S(0)t-alkyl, (17) -C(O)-alkyl,
(18)
Figure imgf000184_0002
(19) S ?
Figure imgf000184_0003
wherein each alkyl is independently selected, (20)
Figure imgf000185_0001
wherein each alkyl is independently selected,
(21)
Figure imgf000185_0002
wherein each alkyl is independently selected,
(22) -N(R4^-C(O)-R48 wherein each R48 is independently selected from the group consisting of: H and aikyl, and
(23) -C(O)-alkyl; each R3, R4, R5, R6 and R7 is independently selected from the group consisting of:
O) H,
(2) aikenyl,
(3) substituted aikenyl,
(4) alkyl, (5) substituted alkyl,
(6) cycloalkyl,
(7) substituted cycloalkyl,
(8) cycloalkylalkyl-,
(9) substituted cycloalkylalkyl-, (10) heterocycloaikyl,
(1 1) substituted heterocycioalkyl,
(12) heterocydoalkyialkyl-,
(13) substituted heterocydoalkyialkyl-,
(14) -C(O)R10, (15) arylheteroaryl-,
(16) substituted arylheteroaryl-,
(17) heteroarylaryl-, (18) substituted heteroaryiaryl-, (19) aryl,
(20) substituted aryl,
(21) heteroaryl, (22) substituted heteroaryl,
(23) heteroarylheteroaryl-,
(24) substituted heteroarylheteroaryi-,
(25) arylaminoheteroaryl-,
(26) substituted aryiaminoheteroaryl-, (27) arylalkynyl-,
(28) substituted arylalkynyl-,
(29) heteroarylalkynyl-,
(30) substituted heteroarylalkynyl-,
(31 ) benzoheteroaryl; wherein said R3, R4, R5, R6 and R7 substituted groups (7), (9), (11), (13), (16),
(18), (20), (22), (24), (26), (28) and (30) are substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NHR20, -N(R20)2 wherein each R20 is independently selected, alkyl, alkenyl, halo, -C(O)-NH-R28, -C(O)OR28, -C(O)R28, and -OR20, wherein said R3, R4, R5, R6 and R7 substituted groups (3) and (5) are substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, halo, -C(O)-NH-R28, -C(O)OR26, and -C(O)R28;
RSA is selected from the group consisting of: halo, -OH, alkyl, -O-alkyl;
R8 is selected from the group consisting of: H, -OH, -N(R10)2) -NR10C(O)R12; each R9 is independently selected from the group consisting of :halogen, -CN,
-NO2, -OR10, -SR10, -N(R10J2, and R10; each R10 is independently selected from the group consisting of: H, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, heterocycioalkyl, heterocycloalkylalkyl, alkylheteroaryl-, alkylaryl-, substituted alkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloaikyl, substituted cycloalkylalkyl, substituted heterocycioalkyl, substituted heterocycloaikylalkyi, substituted alkylheteroaryl-, substituted alkyiaryl-, heterocycloaikenyl
Figure imgf000187_0001
and substituted heterocycloaikenyl, and wherein: said R10 substituted alkyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NHR20, -NO2, -CN, -OR26, halo, -C(O)-NH-R26, -C(O)OR26, and -C(O)R26, and said R10 substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroaryialkyl, substituted cycloalkyl, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycloaikylalkyi, substituted alkylheteroaryl- and substituted alkyiaryl- are substituted with 1 to 3 substituents independently selected from the group consisting of: (1 ) -NH2, (2) -NO2, (3) -CN, (4) -OH, (5) -OR20, (6) -OCF3, (7) alkyl substituted with 1 to 3 independently selected halo atoms, (8) -C(O)R38, (9) alkyl, (10) alkenyl, (1 1 ) haio, (12) -C(O)-NH-R26, (13) -C(O)OR38, (14) -C(O)-NR32-(C(R30)2)n-N(R38)2, (15) -S(O),R3e, (16) -C(O)-NR32-R38, (17) -NR32-C(O)-R38,
Figure imgf000187_0002
(19) -NHR20, (20) cycloalkyl, (21) -O-alkyi-O-R20, (22) hydroxyalkyl, (23) -N(R20J2 wherein each R20 is independently selected, (24) -alky!-OR20, (25) -O-aikyl-OH, (26) -NH (hydroxyalkyl), and (27) oxazolidinone;
R11 is selected from the group consisting of: F, -OH, -CN, -OR10, -NHNR1R10, -SR10 and heteroaryl;
R12 is selected from the group consisting of: alkyl, aryl, heteroaryl, cycioalkyl, cycloalkylalkyl, heterocycloalkyl and heterocycloaikylalkyi; R14 is selected from the group consisting of: aikyl, aryl, heteroaryl, cycloalkyl, cycloalkylalkyl-, heterocycloalkyl, alkylheterocycloalkyl, heterocycloaikylalkyi-, afkylheteroaryt- and alkyiaryl-; R15 is selected from the group consisting of: H, -OH, alkyi, aryl, heteroaryl, cycloalkyl, cycloalkylalkyl-, heterocycloaikyl and heterocycioalkylalkyl-, alkylheteroaryi- and aikylaryl-;
R20 represents alkyl; R23 is selected from the group consisting of: H, alkyl, aryl, cycloalkyl, and cycloalkylalkyl-; each R26 is independently selected from the group consisting of: H and alkyi;
R28 is alkyl; each R30 is independently selected from the group consisting of: H1 alkyl, and F; each R32 is independently selected from the group consisting of: H and alkyl; each R35 is independently selected from the group consisting of: H and Ci to C6 alkyl; each R38 is independently selected from the group consisting of: H, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyi, heterocycloalkylalky], alkylheteroaryl-, alkylaryl-, substituted alkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloalkyl, substituted cycloalkylalkyl, substituted heterocycloaikyl, substituted heterocycioalkylalkyl, substituted alkylheteroaryl- and substituted alkylaryl-, and wherein: said R38 substituted alkyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NO2, -CN, -OR26, halo, -C(O)-NH-R28, -C(O)OR28, and said R38 substituted aryl, substituted aryialkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloaikyl, substituted cycloalkyl alkyi, substituted heterocycloalkyi, substituted heterocycioalkylalkyl, substituted alkylheteroaryl- and substituted alkylaryl- are substituted with 1 to 3 substituents independently selected from the group consisting of: (1) -NH2, (2) -NO2, (3) -CN, (4) -OH, (5) -OR20, (6) -OCF3, (7) -CF3, (8) -C(O)R26, (9) alkyl, (10) alkenyi, (1 1 ) halo, (12) -C(O)-NH-R26, (13) -C(O)OR26, (14) -C(O)-NR32-(C(R30)2)n-N(R26)Ξ) (15) - S(O)1R26, (16) -C(O)N (R32KR26), (17) -NR32C(O)R26,
Figure imgf000189_0001
and (19) -NHR20;
R42 is selected from the group consisting of: alkyi, aryl (e.g., phenyl), heteroaryl, and cycloalkyi; R44 is selected from the group consisting of: H, alkyl, cycloalkyi, and cycloalkylaikyl; and
Each R46 is independently selected from the group consisting of: H, alkyl, cycloalkyi, and cycloalkylaikyl;
Formuia III is described in PCT publication No. 2008/156739, herein incorporated by reference; or c) Formula IV:
Figure imgf000189_0002
IV or the pharmaceutically acceptable salts thereof, wherein: Y1 , Y2, and Y3 are each independently selected from the group consisting of:
-CH=, -N= and -CR9=; z is 1 to 3; Q is a substituent selected from the group consisting of:
Figure imgf000189_0003
(2.5) (2.6 (2.7) (2.8)
(2.9) (2. 0) (2.1 1 ) (2.12)
(2.13) (2.14) (2.15) (2.16)
Figure imgf000190_0001
Each Q1 represents a ring independently selected from the group consisting of: cycloalkyl, substituted cycloalkyl, heterocycioalkyl, substituted heterocycloalkyi, aryl, substituted aryl, heteroaryl, and substituted heteroaryl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R10 moieties; provided that when Q1 is aryl, heteroaryl, substituted aryl or substituted heteroaryl then the carbon atoms at the ring junction are not substituted;
Q2 represents a ring selected from the group consisting of: cycloalkyi, substituted cycloalkyi, heterocycJoalkyi, and substituted heterocycioalkyl, wherein said substituted rings are substituted with 1 to 3 substituents independently selected from the group consisting of: the R10 moieties; Z1 represents -(C(R24)2)w- wherein each R24 is independently selected from the group consisting of: H, alkyl and F, and wherein w is 1 , 2 or 3, and generally w is 1 or 2, and usually w is 1 , and wherein in one example each R24 is H, and in another example w is 1 , and in another example each R24 is H and w is 1 , preferably w is 1 and each R24 is H;
Z2 is selected from the group consisting of: -N(R44)-, -O- and -C(R46J2-; m is 1 to 6; n is 1 to 6; p is 0 to 6; t is 0, 1 , or 2;
R1 is selected from the group consisting of: (I) -CN,
(2) -NO2,
(3) -OR10, (4) -SR10,
(5) -N(R10J2,
(6) R10,
(7) halo,
(8) -CF3; (9) alkenyl;
(10) -C(O)N(R10J2 wherein each R10 is independently selected, and preferably each R10 is independently selected from the group consisting of: (a) H, (b) alkyl, (c) heteroaryl, (d) aryl, and (e) cycloalkyl, wherein for example, each R10 is selected from the group consisting of: H, methyl, butyl, i-propyl, pyridyl, phenyl and cyclopropyl, wherein, for example, said -C(O)N(R10)2 moiety is selected from the group consisting of: -C(O)NH2, -C(O)NH(CH3), -C(O)NH(CH)(CH3)2J -C(O)NH(C4H9), -C(O)NH(C6H5), -C(O)NH(C3H5), and -C(O)NH(C5H4N);
(1 1) arylalkenyl-;
R2 is selected from the group consisting of: (1) H,
(2) -CN,
(3) halo, (4) alkyi,
(5) substituted alkyl wherein said substituted alkyl is substituted with 1 to 3 substitutents selected from the group consisting of: (a) -OH, (b) -O-alkyl (e.g., -0-(Cr C3alkyl), (c) -O-alkyi (e.g., -O-(CrC3alkyl)) substituted with 1 to 3 F atoms, and (d) - N(R40)2 wherein each R40 is independently selected from the group consisting of: (i) H, (ii) CrC3 alkyl and (iii) -CF3,
(6) alkynyl,
(7) alkenyl, <8) -(CH2)mR1\ (9) -N(R26)2,
(1 O) -OR23, (1 1 ) -N(R26)C(O)R42,
(12) cycloalkyl,
(13) cycloalkylalkyl, and (14)
Figure imgf000192_0001
each R3, R4, R5, R6 and R7 is independently selected from the group consisting of:
O) H, (2) alkenyl,
(3) substituted alkenyl,
(4) alkyl,
(5) substituted aikyl,
(6) cycloalkyl, (7) substituted cycloalkyl ,
(8) cycloalkylalkyl-,
(9) substituted cycloalkylalkyl-,
(10) heterocycloalkyl,
(11 ) substituted heterocycloalkyl, (12) heterocycloalkylalkyl-,
(13) substituted heterocycloalkylalkyl-, (14) -C(O)R10 wherein in one example R10 is selected from the group consisting of: alkyl,
(15) arylheteroary1-,
(16) substituted arylheteroaryl-, (17) heteroarylaryi-, such as, for example, pyrimidinylphenyl-, pyrazinylphenyl-, pyridinylphenyl-, furanylphenyl-, thienylphenyl-, and thiazolylphenyl-,
(18) substituted heteroarylaryi-, such as, for example, substituted pyrimidinylphenyl-, substituted pyrazinylphenyl-, substituted pyridinyiphenyl-, substituted furanylphenyl-, substituted thienylphenyl-, substituted thiazolylphenyl-, and substituted pyrimidinylphenyl,
(19) aryl,
(20) substituted aryl,
(21) heteroaryl,
(22) substituted heteroaryl, (23) heteroaryiheteroaryl-,
(24) substituted heteroaryiheteroaryl-,
(25) arylaminoheteroaryl-,
(26) substituted aryiaminoheteroaryl-,
(27) arylalkynyl-, (28) substituted arylalkynyl-,
(29) heteroaryialkynyl-,
(30) substituted heteroaryialkynyl-, (3I) -C(O)NHR28,
(32) cycloaikylheteroarylaryl-, (33) substituted arylaryh
(34) arylalkenylaryh
(35) arylary!-,
(36) substituted arylaikyl-,
(37) arylaikyl-, (38) -SO2aryl,
(39) benzoheteroaryl-C(O)-(substituted heterocycloalkyl)-,
(40) substituted heterocycloalkyl, (41) heterocycloalkyl-C(O)-alky!-, and
(42) benzo[1 ,3]dioxolyl, wherein said R3, R4, R5, R6 and R7 substituted groups (7), (9), (11 ), (13), (16), (18), (20), (22), (24), (26), (28), (30), (33), (36), (39) and (40) are substituted with 1 to 3 substituents independently selected from the group consisting of: -CH2OH, CN, -OH1 -NH2, aikyl, alkenyl, halo, -C(O)-NH-R28, -C(O)NH2, -C(O)OR28, -C(O)R28, -C(alky!)=NOH, -C(alkyi)=NO(alkyl), alkoxy, hydroxy! substituted alkyi, dialkylamine wherein each aikyl group is independently selected, -CF3, -S02alkyl, and -NHC(O)H, wherein said R3, R4, R5, R6 and R7 substituted groups (3) and (5) are substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, halo, -C(O)-NH-R28, -C(O)OR28, and -C(O)R28;
R8 is selected from the group consisting of: H, -OH, aikyl, aryl, -N(R10J2 and -NR1°C(O)R12; each R9 is independently selected from the group consisting of:halogen, -CN, -NO2, -OR10, -SR10, -N(R10J2, and R10; each R10 is independently selected from the group consisting of; H, aikyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkyl aikyl, heterocycloalkyl, heterocycloalkylalkyl, alkylheteroaryl-, alkylaryl-, substituted aikyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloalkyS, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycioalkylalkyl, substituted alkylheteroaryl- and substituted alkylaryl-, and wherein: said R10 substituted aikyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NHR20, -NO2, -CN, -OR26, halo, -C(O)-NH-R26, -C(O)OR26, and -C(O)R26, and said R10 substituted aryl, substituted aryialkyl, substituted heteroaryl, substituted heteroarylalkyi, substituted cycloalkyl, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycloalkylalkyl, substituted alkylheteroaryl- and substituted alkylaryl- are substituted with 1 to 3 substituents independently selected from the group consisting of: (1 ) -NH2, (2) -NO2, (3) -CN,
(4) -OH, (5) -OR20, (6) -OCF3, (7) -CF3, (8) -C(O)R38, (9) aikyl, (10) alkenyl, (1 1 ) halo, (12) -C(O)-NH-R26, (13) -C(O)OR38, (14) -C(O)-NR32-(C(R30)2)π-N(R38)2, (15) -S(O)tR38, (16) -C(O)-NR32-R38, (17) -NR32-C(O)-R38,
Figure imgf000195_0001
(19) -NHR20; R1Ms selected from the group consisting of: F, -OH, -CN, -OR10, -NHNR1R10,
-SR10 and heteroaryl;
R12 is selected from the group consisting of: alkyi, aryl, heteroaryi, cycloalkyl, cyctoalkylalkyl, heterocycloalkyl and heterocycloalkylalkyl;
R14 is selected from the group consisting of: aikyl, aryl, heteroaryl, cycloalkyi, cycloalkylalkyl-, heterocycloalkyl, alkylheterocycloalkyl, heterocycloalkylalkyl-, alkylheteroaryl- and alkyiaryl-;
R15 is selected from the group consisting of: H, -OH, alkyl, aryl, heteroaryl, cycioalkyl, cycloalkylalkyl-, heterocycloalkyl and heterocycloalkylalkyl-, alkylheteroaryl- and alkyiaryl-; R20 represents alkyl;
R23 is selected from the group consisting of: H, alkyl, aryl, cycloalkyl, and cycloalkyialkyl-; each R26 is independently selected from the group consisting of: H and alkyl; R28 is alkyl; each R30 is independently selected from the group consisting of: H, afkyl, and
F; each R32 is independently selected from the group consisting of: H and alky!; each R35 is independently selected from the group consisting of: H and C1 to C6 alkyl; R36 is selected from the group consisting of: H and alkyl; each R38 is independently selected from the group consisting of: H, alkyl, aryl, arylalkyl, heteroaryl, heteroaryl alkyl, cycloalkyS, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, alkylheteroaryl-, alkyiaryl-, substituted alkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyl, substituted cycloaikyl, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycioalkylalkyl, substituted alkylheteroaryl- and substituted alkylaryl-, and wherein: said R38 substituted alkyl is substituted with 1 to 3 substituents independently selected from the group consisting of: -NH2, -NO2, -CN, -OR26, halo, -C(O)-NH-R28 , -C(O)OR28, and -C(O)R28, and said R38 substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalkyi, substituted cycloalkyi, substituted cycloalkylalkyl, substituted heterocycloalkyl, substituted heterocycloalkylaikyl, substituted alkylheteroaryl- and substituted alkylaryl- are substituted with 1 to 3 substituents independently selected from the group consisting of: (1) -NH2, (2) -NO2, (3) -CN, (4) -OH, (5) -OR20, (6) -OCF3, (7) -CF3, (8) -C(O)R26, (9) alkyl, (10) alkenyl, (11) halo, (12) -C(O)-NH-R26, (13) -C(O)OR26, (14) -C(O)-NR32-(C(R30)2)n-N(R26)2, (15) -S(O)1R26, (16) -C(O)N(R32J(R26), (17) -NR32C(O)R26,
Figure imgf000196_0001
(19) -NHR20;
R42 is selected from the group consisting of: alkyl, aryi, heteroaryl, and cycloalkyi;
R44 is selected from the group consisting of: H, alkyl, cycloaikyl, and cycloalkyialkyl; and Each R46 is independently selected from the group consisting of: H, alkyl, cycloalkyi, and cycloalkylalkyi;
Formula IV is described in U.S. publication No. 2007/0232610, herein incorporated by reference.
Non-limiting examples of ERK inhibitors are selected from the group consisting of:
Figure imgf000196_0002
Figure imgf000197_0001
Figure imgf000198_0001
Figure imgf000199_0001
Figure imgf000200_0001
Figure imgf000201_0001
Figure imgf000202_0001
Figure imgf000203_0001
Figure imgf000204_0001
Figure imgf000205_0001
Figure imgf000206_0001
Figure imgf000207_0001
Figure imgf000208_0001
Figure imgf000210_0001
Figure imgf000211_0001
Figure imgf000212_0001
Figure imgf000213_0001
Figure imgf000214_0001
Figure imgf000215_0001
Figure imgf000216_0001
Figure imgf000217_0001
Figure imgf000218_0001
Figure imgf000219_0001
Figure imgf000220_0001
Figure imgf000221_0001
Figure imgf000222_0001
Figure imgf000223_0001
Figure imgf000224_0001
Figure imgf000225_0001
Figure imgf000226_0001
Figure imgf000227_0001
Figure imgf000228_0001
Figure imgf000229_0001
Figure imgf000230_0001
or a pharmaceutically acceptable isomer, sa!t, solvate or co-crystal form thereof.
These compounds are described in PCT publication No. WO 2007/070398, U.S. application No. US 61/030407, PCT publication No. WO 2008/156739 and U.S. publication No. 2007/0232610, herein incorporated by reference. The pharmacoiogical properties of the compounds of this invention may be confirmed by a number of pharmacological assays. The exemplified pharmacological assays which are described herein below have been carried out with compounds according to the invention and their salts, solvates, esters or prodrugs.
This invention is also directed to pharmaceutical compositions which comprise at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound and at least one pharmaceutically acceptable carrier. For preparing pharmaceutical compositions from the compounds described by this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. The powders and tablets may be comprised of from about 5 to about 95 percent active ingredient. Suitable solid carriers are known in the art, e.g., magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and methods of manufacture for various compositions may be found in A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co., Easton, Pennsylvania.
Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection or addition of sweeteners and opacifiers for oral solutions, suspensions and emulsions. Liquid form preparations may also include solutions for intranasal administration.
Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas, e.g. nitrogen. Also included are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.
The compounds of the invention may also be deliverable transdermally. The transdermal compositions can take the form of creams, lotions, aerosols and/or emulsions and can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose.
The compounds of this invention may also be delivered subcutaneously. Preferably the compound is administered orally or intravenously. Also contemplated are delivery methods that are combinations of the above- noted delivery methods, Such methods are typically decided by those skilled in the art.
Preferably, the pharmaceutical preparation is in a unit dosage form. In such form, the preparation is subdivided into suitably sized unit doses containing appropriate quantities of the active component, e.g., an effective amount to achieve the desired purpose.
The quantity of active compound in a unit dose of preparation may be varied or adjusted from about 1 mg to about 100 mg, preferably from about 1 mg to about 50 mg, more preferably from about 1 mg to about 25 mg, according to the particular application.
The actual dosage employed may be varied depending upon the requirements of the patient and the severity of the condition being treated. Determination of the proper dosage regimen for a particular situation is within the skill of the art. For convenience, the total daily dosage may be divided and administered in portions during the day as required.
The amount and frequency of administration of the compounds of the invention and/or the pharmaceutically acceptable salts thereof will be regulated according to the judgment of the attending clinician considering such factors as age, condition and size of the patient as well as severity of the symptoms being treated. A typical recommended daily dosage regimen for oral administration can range from about 1 mg/day to about 500 mg/day, preferably 1 mg/day to 200 mg/day, in two to four divided doses.
Another aspect of this invention is a kit comprising a therapeutically effective amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound and a pharmaceutically acceptable carrier, vehicle or diluent.
Yet another aspect of this invention is a kit comprising an amount of at least one compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester or prodrug of said compound and an amount of at least one anticancer therapy and/or anti-cancer agent listed above, wherein the amounts of the two or more ingredients result in desired therapeutic effect.
The invention disclosed herein is exemplified by the following preparations and examples which should not be construed to limit the scope of the disclosure. Alternative mechanistic pathways and analogous structures will be apparent to those skilled in the art. Where LC/MS data are presented, analyses was performed using an Applied
Biosystems API-100 mass spectrometer and Shimadzu SCL-10A LC column: Altech platinum C18, 3 micron, 33mm x 7mm ID; gradient flow: 0 min - 10% CH3CN, 5 min -
95% CH3CN, 7 min - 95% CH3CN, 7.5 min - 10% CH3CN1 9 min - stop. The retention time and observed parent ion are given.
The following solvents, reagents and reaction conditions may be referred to by their abbreviations:
Aq: aqueous g: grams psi: pounds per square inch pH: percent Hydrogen
°C: degrees Celsius h: hours
THF: Tetrahydrofuran HMDS:
Et2O: diethyloxide
SEM: 2-(trimethylsilyi)ethoxymethyl
LC-MS: Liquid chromatography mass spectrometry
DCM: dichloromethane N: Normal mS: milliliter
NBS: N-Bromosuccinimide it: room temperature
MeOH: methanol DIEA: diisopropylethylamine
EtOAc: ethyl acetate
EtOH: ethanol
DMF: dimethylformamide
WT%: weight percent m/z: mass per charge
LiOH: lithium hydroxide
DMSO: dimethylsulfoxide HPLC: high performance liquid chromatography
Ret: retention
RP: reverse phase
CH3CN: acetonitriie MeCN: acetonitriie pTSA: para-to!uene sulfonic acid
RT: retention time
NaOH: sodium hydroxide
CDI: N,N'-carbonyldiimidazole mg: milligram
PMA: phosphomolybdic acid
CO2: carbon dioxide
LiHMDS: lithium hexamethyldisilazane
HMDS: hexamethyldisϋazane Pd/C: palladium on carbon
H2: hydrogen gas
NIS: N-iodosuccinimide
PDCI2(dppf): [1 ,1 '-bis(diphenylphosphino)ferrocene] dichloropalladium(il) μmol: micromole TFA: trifluoroacetic acid
NMP: N-methyl-2-pyrrolidone min: minute
NaIO4: sodium periodate
DME: dimethylethane OsO4: osmium tetroxide
Na2S2Og: sodium thiosulfate
AcOH: acetic acid
NaBH3CN: sodium cyanoborohydride
H2O: water BBN: 9-borabicyclo[3.3.1]nonane
CH2CI2: dichloromethane
BOC: tertiary-butyloxycarbonyi POCI3: phosphorous oxychloride
NaHCO3: sodium bicarbonate
NH4CI: ammonium chloride
Na2SO4: sodium sulphate HC!: hydrogen chloride
M: Molar mmol: miilimolar
NH3: ammonia
DIEA; diisopropylethylamine Bu3SnCN: tributyltin cyanide
Pd[P(t-Bu)3]2: bis(tributyl)Phosphine) palladium
Pd(PPh3)4: tetrakis(triphenylphosphine) palladium
K2CO3: potassium carbonate
EDC: 1 -ethyl-3-(3-dimethylaminopropyl)-carbodiimide UV: ultraviolet
K3PO4: potassium phosphate
LDA: lithium diisopropylamide
Tf: trifluoromethanesulfonyl
NaH: sodium hydride
Figure imgf000235_0001
Scheme 1
Pvridine-2.5-dicarboxvlic acid 2-tert-butv) ester 5-methvi ester
Figure imgf000236_0001
5-{Methoxycarbony!)pyridine-2-carboxylic acid (7.72 g, 42.65 mmol) was suspended in tert-butanol {70 mL) and pyridine (25 ml_) and cooled in an ice-water bath. 4- Toluenesulfonyl chloride (19.4 g, 102 mmol) was added in one portion and the mixture was stirred 30 minutes in the ice-water bath. The reaction mixture was warmed to room temperature and stirred for 16 h. The reaction mixture was then slowly poured into a stirring mixture of saturated aqueous sodium bicarbonate (300 mL) and ethyl ether (150 mL). The resulting two-phase mixture was then extracted with ethyl ether (3x150 mL). The extracts were combined, washed with brine, dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product (5.8 g, 51% yield) was used in the next step without further purification.
Piperidine-2,5-dicarboxylic acid 2-tert-butyl ester 5-methyl ester
Figure imgf000236_0002
Pyridine-2,5-dicarboxylic acid 2-tert-buty! ester 5-methyl ester (5.8 g, 24.45 mmol) was dissolved in glacial acetic acid (30 mL) and hydrogenated at 50-60 psi for 3 days with 10% palladium on carbon catalyst (0.6 g). The reaction mixture was filtered through a pad of Ceiite which was then washed with methanol. The filtrates were combined and concentrated under reduced pressure. The residue was dissolved in water (100 mL) and solid sodium carbonate (15 g) was added to bring the pH to 8. The solution was extracted with dichloromethane (2x100 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound (5.2 g, 87%).
Piperidine-1 ,2,5-tricarboxylic acid 1 ,2-di-tert-butyl ester 5-methyl ester
Figure imgf000237_0001
Piperidine-2,5-dicarboxyiic acid 2-tert-butyl ester 5-methyl ester (9.0 g, 37.0 mmol) and triethylamine (15.5 ml, 111 mmol) were combined in dichloromethane (70 ml_) and cooled to 0 °C. Di-tert-butyldicarbonate (12.2 g, 55.5 mmol) was added and reaction mixture was warmed to room temperature and stirred for 16 h. The reaction mixture was diluted with water (100 ml_) and extracted with dichloromethane (2x100 ml_). The combined extracts were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by Biotage chromatography column (hexanes/EtOAc, 9:1 to 8:2). A colorless oil was obtained (12.6 g, 99%).
1.6-bisftert-butoxycarbonyl)piperidine-3-carboxylic acid
Figure imgf000237_0002
Piperidine-1 ,2,5-tricarboxylic acid 1,2-di-tert-butyl ester 5-methy! ester (36.7 mmol, 12.6 g) was dissolved in 180 mL of 2:1 THF:H2O. To this solution was added 2N sodium hydroxide solution (52 mL) and the reaction mixture was stirred for 16h at room temperature. The reaction mixture is acidified to pH = 3 with 1 N HCl(aq) and extracted with dichloromethane(3x100 mL). The combined organics are then dried over Na2SO4 and the solvent removed in vacuo to yield the title compound (12.0 g, 100% yield).
Di-tert-butyl 5-(3-ethoxy-3-oxopropanov!)piperidine-1,2-dicarboxvlate
Figure imgf000238_0001
i jδ-bisttert-butoxycarbonylJpiperidine-3-carboxylic acid (41.4 mmol, 13.6 g) and N, N'- carbonyldiimidazole (CDI) (51.75 mmol, 8.4 g) in anhydrous THF (140 mL) were stirred 16 h at room temperature under argon. In a separate, sealed and argon- flushed flask, lithium HMDS (1.0 M in THF, 86.9 mmol) is added to 80 mL anhydrous THF stirring at -78° C. To this solution is added dropwise anhydrous ethyl acetate (89 mmol, 8.69 mL). This solution is allowed to stir at -78° C for 1 hour prior to the dropwise addition of the original CDi/acid solution that had been stirred overnight. The reaction mixture is allowed to stir and warm to room temperature overnight. The reaction is then quenched with saturated NH4C!(aq) (250 mL) and extracted with Et2O (100x2). The combined organics are then washed with water, saturated brine, and dried over Na2SO4. The solvent is removed in vacuo and the residue (12.6 g) was used in the next step without further purification.
Figure imgf000238_0002
Scheme 2
Di-tert-butyl 5-(7-hydroxypyrazoio[1 ,5-alpyrimidin-5-vθpiperidine-1 ,2-dicarboxylate
Figure imgf000239_0001
Di-tert-butyl 5-(3-ethoxy-3-oxopropanoyl)piperidine-1,2-dicarboxylate (18.79 mmol, 7.5
Figure imgf000239_0004
g) and 3-amino-1 H-pyrazole (15.0 mmol, 1.25g) were mixed and heated neat at 100° C for 16 hours. The resulting residue is dissolved in dichloromethane (100 ml.) and concentrated in vacuo to remove water formed during cyclization. LC-MS: 419 [M+H], This solid is taken forward without further purification.
Di-tert-butyl 5-(7-chloropyrazolori ,5-a1pyrimidin-5-yl)piperidine-1 ,2-dicarboxyiate
Figure imgf000239_0002
Di-tert-butyl 5-(7-hydroxypyrazo!o[1 ,5~a]pyrimidin-5-yl)pipeπdine-1 ,2-dicarboxylate {16.7 mmol, 7.0 g), N, N'-diisopropyiethylamine (51.9 mmol, 9.0 ml) and POCi3 (80 mL) were mixed and stirred at room temperature for 18 hours. The solution is concentrated in vacuo and cooled to 0° C in an ice bath. The reaction is then quenched with sat. NaHCO3(aq) and extracted with DCM (100 ml_x3). The combined organics are dried with Na2SO4 and the solvent removed in vacuo. LC-MS: 437 [M+H]. The resulting oil was used in the next step without further purification.
Di-tert-butyl 5-(7-aminopyrazoloH ,5-a1pyrimidin-5-yl)piperidine-1 ,2-dicarboxylate
Figure imgf000239_0003
Di-tert-butyl 5-(7-ch!oropyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 ,2-dicarboxylate was dissolved in 30 mL ~7N ammonia in methanol in a sealed vessel. The reaction mixture was heated at 80° C for 16 hours. After 16 hours, the reaction mixture is cooled to room temperature and concentrated in vacuo to yield a brown solid. LC-MS: 418 [M+H]. The title compound was used in the next step without further purification.
Di-tert-butyl 5-(7-(bisff2-(trimethylsiiyl)ethoxy)methyl)amino)pyrazoloπ .5-aipyrimidin- 5-yl)piperidine-1 ,2-dicarboxvlate
Figure imgf000240_0001
Di-tert-butyl 5-(7~aminopyrazolo[1 ,5-a]pyrimidin-5-yi)piperidine-1 ,2-dicarboxylate (5.5 g, 13.18 mmol) was dissolved in 1 ,2-dichloroethane {60 ml_). To this solution was added N, N'-diisopropylethylamine (92.28 mmol, 16.1 ml_). The resulting solution was stirred at room temperature while 2-(tritnethylsilyi)ethoxymethyl chloride (SEM-CI, 8.14 mL, 46.14 mmol) was added dropwise. After the addition is completed, the reaction mixture was stirred at 90° C for 2 h. The solvent is removed in vacuo and the residue is purified on silica gel column (0% to 60% ethyl acetate in hexanes gradient) to yieid the title compound (3.8 g, 34% yield over three steps) as pale yellow oil. LC-MS: 678 [M+H].
Di-tert-butyl 5-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolori ,5- aipyrimidin-5-yl)piperidine-1 ,2-dicarboxylate
Figure imgf000240_0002
N-lodosuccinimide (1.26 g, 5.61 mmol, dissolved in 25 ml acetonitrile) was added into a solution of di-tert-buty! 5-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)piperidine-1 ,2-dicarboxylate (3.8 g, 5.61 mmol) in acetonitrile (26 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated in vacuo and purified via silica gel chromatography (0% to 30% ethyl acetate in hexanes gradient) to yield the title compound (3.16 g, 70% yield) at clear oil. LC-MS: 804 [M+H].
Di-tert-butyl 5-(7-(bisf(2-(trimethylsilyl)ethoxy)methyl)amino)-3-(Quinolin-3- yl)pyrazoloM ,5-a]pyrimidin~5-yl)piperidine-1 ,2-dicarboxylate
Figure imgf000241_0001
3-Quinoline boronic acid (1.25 mmol, 216 mg), K3PO4 (1.87 mmol, 400 mg), and PdCI2(dppf) • CH2CI2 (0.062 mmol, 51 mg) was added to a solution of di-tert-butyl 5-(7- (bis((2-(trimethytsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yi)piperidine-1 ,2-dtcarboxylate (0.62 mmol, 500 mg) in dioxane (5 ml_). To this suspension was added distilled H2O (0.5 ml_). The reaction mixture was stirred at 100° C under an argon atmosphere for 18 hours. The reaction mixture was concentrated in vacuo and then purified via silica gel chromatography (0% to 60% ethyl acetate in hexanes gradient) to yield the title compound (450 mg, 90% yield) as yellow oil. LC-MS: 805 [M+H].
5^-amino-3-(quinolin-3-yl)pyrazolofi .5-aipyrimidin-5-yl)piperidine-2-carboxyiic acid
Figure imgf000241_0002
Di-tert-buty! 5-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)pipehdine-1 ,2-dicarboxylate (45 mg, 0.056 mmol) was dissolved in ethanol (1 ml) and treated with 3N hydrochloride solution (1.4 ml) at 65 °C for 4 h. The reaction solution was concentrated and purified by prep-LC to afford the title compound (9.6 mg): LC/MS RT = 2.01 min. Mass calculated for, M+H 389.17, observed 389.17.
By essentially the same procedure given in Scheme 2, the compounds listed in Table 1 can be prepared.
Table 1
Figure imgf000242_0001
Figure imgf000243_0001
Figure imgf000244_0001
Figure imgf000245_0001
Figure imgf000246_0002
Figure imgf000246_0001
Scheme 3 Methyl 5-(7-amino-3-(quinoiin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-2-carboxylate
Figure imgf000247_0001
Di-tert-butyl 5-(7-(bis({2-(trimethylsilyi)ethoxy)m8thyl)amino)-3-(quino!in-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1,2-dicarboxylate (370 mg, 0.46 mmol) in methanol (12.6 mL) was treated with 4N hydrogen chloride in dioxane (5.4 ml). The reaction mixture was heated at 80 °C for 16 h, cooled to room temperature and concentrated in vacuo. The residue was purified by prep-LC to afford the title compound: LC/MS RT = 2.31 min. Mass calculated for, M+H 403.18, observed 403.18.
Methyl 5-(7-amino-6-bromo-3-(αuinoiin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)pipeπdine-2- carboxylate
N-bromosuccinimide (44 mg, 0.249 mmol) was added into a solution of methyl 5-(7- amino-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-2-carboxy!ate (100 mg, 0.249 mmol) in acetonitrite (1.5 mL) and methanol (0.5 ml). The resulting solution is stirred at room temperature for 1 hour. The reaction was concentrated in vacuo and purified by prep-LC to afford the title compound (22.32 mg): LC/MS RT = 2.56 min. Mass calculated for, M+H 481.09, observed 481.09.
Figure imgf000247_0003
5-(y-amino-e-bromo-3-(quinoiin-a-yl)pyrazolofi ^-aiPyrimiciin-5-yl)pipericiine-2- carboxylic acid
Methyl 5-(7-amino-6-bromo-3-(quinoiin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-2- carboxylate (1 1 mg, 0.023 mmol) in 2:1 THFiH2O (1.5 ml_) was treated with 2N NaOH(aq) (0.25 ml_). The resulting solution was stirred at room temperature for 18 hours. This solution is reduced in vacuo and purified by prep-LC to afford the title compound (8.9 mg): LC/MS RT = 2.16 min. Mass calculated for, M+H 467.08, observed 467.08.
By essentially the same procedure given in Scheme 3, the compounds listed in Table 2 can be prepared.
Table 2
Figure imgf000248_0001
Figure imgf000249_0001
Figure imgf000250_0003
Figure imgf000250_0001
Scheme 4 5-(7"Amino-6-bromo-3-quinoiin-3-yl-pyrazoiof1 ,5-a1pyrjmidin-5-yl)-1 -(2-methoxy- acetyl)-piperidine-2-carboxylic acid
A mixture of 5-(7-amino-6-bromo-3-quinolin-3-yl-pyrazolo[1 ,5-a]pyrimidin-5-yi)- piperidine-2-carboxylic acid methyl ester (20 mg, 0.042 mmol), methoxy-acetyl chloride (13 mg, 0.071 mmol) and triethylamine (0.024 ml, 0.167 mmol) in THF:DMF (0.5 ml:0.5 ml) was stirred at room temperature for 16 h. The crude mixture was treated with 2.0 N sodium hydroxide solution (0.14 ml) at room temperature for 16 h. The reaction mixture was acidified with 1.0 N aqueous hydrogen chloride solution (0.5 ml), concentrated and purified by prep-LC to afford the title compound (7.94 mg): LC/MS RT = 3.06 min. Mass calculated for, M+H 539.10, observed 539.10. By essentially the same procedure given in Scheme 4, the compounds listed in Table 3 can be prepared.
Table 3
Figure imgf000250_0002
Figure imgf000251_0001
Figure imgf000252_0001
Figure imgf000253_0001
Figure imgf000254_0001
Scheme 5
4-(2-Ethoxycarbonyl-acetylVcvclohexanecarboxyiic acid methyl ester
Figure imgf000254_0002
Cyclohexane-1 ,4-dicarboxylic acid monomethyl ester (26.9 mmol, 5.0 g) and N, N'- carbonyldiimidazole (33.6 mmol, 5.44 g) in anhydrous THF (80 ml_) were stirred 16 h at room temperature under argon. In a separate, sealed and argon-flushed flask, lithium HMDS (1.0 M in THF1 56.5 ml) is added to 40 mL anhydrous THF stirring at - 78° C. To this solution is added dropwise anhydrous ethyl acetate (57.8 mmol, 5.65 mL). This solution is allowed to stir at -78° C for 1 hour prior to dropwise addition of original CDI/acid solution that had been stirring overnight. The reaction mixture was allowed to stir and warm to room temperature overnight. The reaction is then quenched with saturated NH4CI(aq) (200 mL) and extracted with Et2O (100x2). The combined organics are then washed with water, saturated brine, and dried over Na2SO4. The solvent was removed in vacuo and the crude residue (8.5 g) was used in the next step without further purification.
4-(7-Hvdroxy-pyrazoloh ,5-aipyrimidin-5-yl)-cvclohexanecarboxylic acid methyl ester
Figure imgf000255_0001
4-{2-Ethoxycarbony[-acetyl)-cyc!ohexanecarboxylic acid methyl ester (8.5 g) and 3- amino-1 H-pyrazole (26.5 mmol, 2.2 g) were mixed and heated neat at 100° C for 16 hours. The resulting residue is dissolved in dichloromethane (100 mL) and concentrated in vacuo to remove water formed during cyciization. This solid is taken forward without further purification. LC-MS: 403 [M+HJ.
4-(7-Chloro-pyrazolo[1 ,5-aipvrimidin-5-vl)-cyclohexanecarboxvlic acid methyl ester
Figure imgf000255_0002
4-(7-Hydroxy-pyrazolo[1 ,5-ajpyrimidin-5-yl)-cyciohexanecarboxylic acid methyl ester (10.6 g), N, N-dimethylaniiine (15.0 mi) and POCI3 (120 mL) were mixed and stirred at room temperature for 18 hours. The solution is reduced in vacuo and cooled to 0° C in ice bath. The reaction is then quenched with sat. NaHCO3(aq) and extracted with DCM (100 mLx3). The combined organics are dried with Na2SO4 and the solvent removed in vacuo. The resulting oil was purified via silica gel column on 20% to 100% ethyl acetate in hexanes gradient to yield the titled compound (2.5 g, 32% over three steps), LC-MS: 294 [M+H].
4-(7-Amino-pyrazolori ,5-a1pyrimidin-5-yl)-cvciohexanecarboxylic acid methyl ester
Figure imgf000255_0003
4-(7-Chloro-pyrazolo[1 ,5-a]pyrimidin-5-yl)-cyclohexanecarboxylic acid methyl ester (2.5 g, 8.51 mmol) was dissolved in 15 mL ~7N ammonia in methanol in a sealed vessel. The reaction mixture was heated at 80° C for 16 hours. After 16 hours, the reaction mixture is cooled to room temperature and concentrated in vacuo to yield a brown solid (2.58 g). The title compound was used in the next step without further purification. LC-MS: 275 [M+H].
4-(7-|-Bis-(2-trimethylsilanyl-ethoxymethyl)-amino1-pyrazolo[1 ,5-aiPyrimidin-5-yll- cvclohexanecarboxylic acid methyl ester
Figure imgf000256_0001
4-(7-Amino-pyrazolo[1 ,5-a]pyrimidin-5-yl)-cyclohexanecarboxylic acid methyl ester (2.58 g, 9.4 mmoi) was dissolved in 1 ,2-dichloroethane (20 mL). To this solution was added N, N'-diisopropylethylamine (65.8 mmol, 11.5 mL). The resulting solution was stirred at room temperature while 2-(trimethylsilyl)ethoxymethyl chloride (SEM-CI, 5.8 mL, 33.0 mmol) was added dropwise. After the addition was completed, the reaction mixture was stirred at 90° C for 2 h. The solvent is removed in vacuo and the residue is purified on silica gel column (0% to 60% ethyl acetate in hexanes gradient) to yield the title compound (2.05 g, 41% yield over two steps) as pale yellow oil. LC-MS: 535 [M+H].
4-f7-rBis-(2-trimethylsilanyl-ethoxymethyl)-aminol-3-iodo-pyrazoloπ .5-aipyrimidin-5- yl)-cvclohexanecarboxylic acid methyl ester
Figure imgf000256_0002
N-lodosuccinimide (81 1 mg, 3.6 mmol) was added into a solution of 4~{7-[bis-(2- trimethylsilanyl-ethoxymethyl)-amino]-pyrazolo[1 ,5-a]pyrimid!n-5-yl}- cyclohexanecarboxylic acid methyl ester (1.75 g, 3.27 mmol) in acetonitrile (25 mL). The resulting reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated in vacuo and purified via silica gel chromatography (0% to 30% ethyl acetate in hexanes gradient) to yield the title compound (1.79 g, 82.8% yield) at clear oil. LC-MS: 661 [M+H].
4-{7-rBis-(2-tπmethylsilanyl-ethoxymethyl)-amino1-3-quinolin-3-yl-Pyrazolo[1 ,5- aipyrimidin-5-yll-cvclohexanecarboχylic acid methyl ester
Figure imgf000257_0001
3-Quinoline boronic acid (2.42 mmol, 420 mg), K3PO4 (3.63 mmol, 771 mg), and PdCI2(dppf) ■ CH2CI2 (0.121 mmol, 100 mg) was added to a solution of 4-{7-[Bis-(2- trimethylsilanyi-ethoxymethyl)-amino]-3-iodo-pyrazolo[1 ,5-a]pyrimidin-5-yl}- cyclohexanecarboxylic acid methyl ester (1.21 mmoi, 800 mg) in dioxane (10 mL). To this suspension was added distilled H2O (1.0 mL). The resulting reaction mixture was stirred at 100° C under an argon atmosphere for 18 hours. The reaction mixture was concentrated in vacuo and then purified via silica gel chromatography (0% to 60% ethyl acetate in hexanes gradient) to yield the title compound (630 mg, 79% yield) as yellow oil. LC-MS: 662 [M+H].
4-(7-[Bis-(2-trimethylsilanyl-ethoxymethyi)-amino1-6-bromo-3-quinolin-3-yl- pyrazolof^5-alpyrimidin-5-yll-cvciohexanecarboxylic acid methyl ester
Figure imgf000257_0002
N-bromosuccinimide (148 mg, 0.832 mmol) was added to a solution of 4-{7-[bis-(2- trimethy!silanyl-ethoxymethyl)-amino]-3-quinolin-3-yl-pyrazolo[1 ,5-a]pyrimidin-5-yl}- cyciohexanecarboxylic acid methyl ester (500 mg, 0.756 mmol) in acetonitrile (10 ml_). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo and the resulting oil is then purified via silica gel chromatography (0% to 30% ethyl acetate in hexanes gradient) to yield the title compound (530 mg, 95% yield) as yellow oil. LC-MS: 740 [M+H].
4-(7-Amino-6-bromo-3-quinolin-3-yl-pyrazoloπ .5-alpyrimidin-5-yl)- cyclohexanecarboxylic acid
Figure imgf000258_0001
4-{7-[Bis-(2-trimethylsilanyl-ethoxymethyl)-amino]-6-bromo-3-quinolin-3-yl- pyrazolo[1 ,5-a]pyrimidin-5-yl}-cyctohexanecarboxylic acid methyl ester (100 mg, 0.135 mmol) in 2:1 MeOH :H2O (3 mL) was treated with 2N NaOH(aq) (0.5 mL). The reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was treated with 1 N hydrochloride solution (2 ml) at 65 °C for 4 h. The reaction mixture was concentrated and purified by prep-LC to afford the title compound (21.8 mg): LC/MS RT = 3.57 min. Mass calculated for, M+H 466.08, observed 466.08. By essentially the same procedure given in Scheme 5, the compounds listed in Table 4 can be prepared.
Table 4
Figure imgf000258_0002
Figure imgf000259_0001
Figure imgf000260_0001
Figure imgf000261_0001
Following additional compounds in table~4a were synthesized following the general experimental procedures described in scheme-5
TABLE-4A
Figure imgf000261_0002
Figure imgf000262_0001
Figure imgf000263_0001
Figure imgf000264_0001
Figure imgf000265_0001
Figure imgf000266_0001
Figure imgf000267_0001
Figure imgf000268_0001
Figure imgf000269_0001
Figure imgf000270_0001
Figure imgf000271_0001
Figure imgf000272_0001
Figure imgf000273_0001
Figure imgf000274_0001
Figure imgf000275_0001
Figure imgf000276_0001
Figure imgf000277_0001
Figure imgf000278_0002
SCHEME- 5a
Figure imgf000278_0001
6-(7-amino-3-(quinolin-3-yl)pyrazok>[1 ,5-a]pyrimidin-5-yl)hexanoic acid (4.87)
To a solution of methyl hex-5-enoate (12 mg, 0.1 mmol) in THF (0.5 ml) under argon at 0 °C was added 9-BBN {0.2 mmol, 0.4 mi of 0.5 M solution in THF). The mixture was warmed to room temperature and stirred for 16 h. Potassium phosphate (3 M -in H2O, 0.2 mmoi) was added followed by the addition of 5-chloro-3-(quinolin-3-yl)-N,N- bis((2-(trimethylsily!)ethoxy)methyl) pyrazo!o[1 ,5-a]pyrimidin-7-amine (50 mg, 0.09 mmol) in DMF (0.5 ml) and Pd(dppf)CI2-CH2CI2 (5 mg, 0.005 mmol). The reaction mixture was heated at 90 °C for 16 h under argon. The solution was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 2:1 THFiMeOH (3 ml_) and was treated with 2N NaOH(aq) (0.25 ml_). The resulting solution was stirred at room temperature for 4 hours followed by the addition of 2N hydrochloride solution (1.0 ml). The solution was heated at 65 °C for 2 h, cooled to room temperature, concentrated and purified by prep-LC to afford the title compound (8.2 mg): LC/MS RT = 2.74 min. Mass calculated for, M+H 376.17, observed 376.17. Analogues to the compound 4.87, following compounds in the table-4C can be synthesized as described above in the scheme-5a
Table-4C
Figure imgf000279_0001
Figure imgf000280_0001
Figure imgf000281_0001
Following nucleophilic displacement reaction of 5-chloro-3-(quinolin-3-yl)-N,N-bis{(2- (trimethylsilyl)ethoxy)methyl) pyrazolo[1 ,5-a]pyrimidin-7-amine with appropriate amine or alchol ( experimental procedure described in Scheme-5a, compounds in the table- 4C can be synthesized. Synthesis of 2-methyl-6-(4.4.5.5-tetramethyl-1.3.2-dioxaborolan-2-vnthienor3,2- bfoyridine:
Figure imgf000282_0001
Part A:
To a stirring solution of thiophenecarboxyfic acid (1 equiv) in DMF and f-BuOH at 0 °C was added the DPPA (1 equiv) and TEA (1 equiv). The reaction was then stirred at room temperature for 1 hour, then warmed to reflux and stirred for 15 hours. The reaction was allowed to cool to room temperature, and then evaporated to dryness.
The resulting residue was dissolved in EtOAc then washed with sat. aq. NH4CI, sat. aq. NaHCO3, and brine, dried over sodium sulfate, filtered, and purified on silica gel to afford the product.
Part B:
A solution of HCI in dioxane was added to a stirring solution of the product from Part A in methanol at 0 °C. The reaction was then stirred at reflux for 1 hour, then allowed to cool to room temperature and concentrated to afford the product.
Part C: Bromomalonaldehyde (1 equiv) was added to a solution of the product from Part B (1 equiv) in glacial acetic acid at room temperature. The mixture was stirred at reflux for 1 hour, then allowed to cool to room temperature, then evaporated to dryness. The residue was dissolved in methanol, filtered through a pad of celite, concentrated, and purified on silica gel to afford the product.
Part D:
A mixture of the product from Part C (1 equiv), bispinacolatodiboron (1.2 equiv), PdCbdppf (0.1 equiv), and potassium acetate (3 equiv) in dioxane was heated at 85 °C until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction was allowed to cool to room temperature, filtered through a pad of celite, concentrated to dryness, treated with diethyl ether, filtered through a pad of celite, then concentrated to afford the desired product, 2-methyl-6-{4,4,5l5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)thieno[3,2-b]pyridine.
Synthesis of 6-(4.4.5.5-tetramethyl-1.3.2-dioxaborolan-2-yl)thienof3,2-biPyridine:
Essentially utilizing the procedure described above scheme- the title compound 6- (4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)thieno[3,2-b]pyridine can be synthesized
P N
Figure imgf000283_0001
SCHEME-7
Synthesis of 5-(4,4.5.5-tetramethyl-1 ,3,2-dioxaboroian-2-vhfurof2.3-bipyridine:
(modified procedure from the patent # GB2289276A)
H — TMS
Figure imgf000283_0002
Part A:
To a solution of 5-bromo-2-(1H)-pyridone (18.8 gf 10.8 mmol) in dry toluene {400 ml_) was added N-iodosuccinimide (24.3 g, 10.8 mmol) under nitrogen. The reaction mixture was stirred at 90 °C for 20 minutes then cooled to 25 °C. The precipitate was filtered off and washed with methanol, dried under vacuum to give the pink-orange color solid 28.0 g (86%). 1H NMR (300 MHz, CDCi3 ) δ 8.1 (s, 1 H), 7.6 (s, 1 H). Mass caicuiated for formula C5H3BrINO 298.84, observe MS ES+: 300.0/302.0. Part B:
The iodide (6.0 g, 20.0 mmol), Pd(OAc)2 (45 mgt 0.2 mmol), PPh3 (105 mgt 0.4 mmol), CuI (76 mg, 0.4 mmol) were added to a round bottom flask under nitrogen. Dry THF (30 ml_) was added and the solution was degassed 2 minutes with nitrogen. TMS- acetylene (2.9g, 30 mmol) was added followed with n-BuNH2 (2.9 g, 40 mmol). The homogeneous green solution was heated to 38 °C for 4 hours. The reaction mixture was cool to 25 °C, concentrated to dryness and then dissolved in EtOAc (100 ml_). The solution was washed with saturated sodium potassium tartrate (50 ml_), followed with 0.1 N HCI (50 mL), saturated NaHCO3 (50 mL), and brine (5OmL), dried over NasSO4. Purification by column chromatography (SiO2, 50% ethyl acetate/hexanes) afforded compound as a green solid 4.40 g (81 %). 1H NMR (300 MHz, CDCI3) δ 7.85 (s, 1 H), 7.5 (s, 1 H), 0.2 (s, 9H).
Part C: To a EtOH (8.0 mL) solution of compound from Part B (1.12 g, 4.1 mmol) was added CuI (39 mg, 0.2 mmol) and followed with Et3N (3.5 mL) the resulting mixture was stirred at 70 °C for 3 hours. The reaction mixture was allowed to cool to 25 °C, concentrated to dryness and then dissolved in toluene (3OmL). The solution was washed with 0.1 N HCI (20 mL), saturated NaHCO3 (20 mL), and brine (30 mL), dried over Na2SO4. Purification by column chromatography (SiO2, 20% ethyl acetate/hexanes) afforded compound as a white solid 0.26 g (32%). H NMR (300 MHz, CDCl3) δ 8.4 (s, 1 H), 8.1 (s, 1H), 7.7 (s, 1H), 6.8 (s, 1 H).
Part D: A mixture of the product from Part C (1 equiv), bispinacoiatodiboron (1.2 equiv), PdCI2dppf (0.1 equiv), and potassium acetate (3 equiv) in dioxane was heated at 85 °C until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction was allowed to cool to room temperature, filtered through a pad of celite, concentrated to dryness, treated with diethyl ether, filtered through a pad of celite, then concentrated to afford the desired product, 5-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)furo[2,3-b]pyridine.
2-methyl-5-(4.4.5.5-tetramethyi-1 ,3,2-dioxaborolan-2-yl)furo[2,3-blPyridine:
I- Λ ,
YV
Figure imgf000284_0001
Part B
SCHEME-8
Part A: A mixture of 5-bromo-3-iodopyridin-2-ol (1 equiv), copper iodide (3 equiv), palladium catalyst (0.05 equiv), TBAF (1 equiv), triethylamine (3.3 equiv) and alkyne (1 equiv) in toluene was stirred at room temperature until the reaction was deemed complete by TLC analysis. The reaction mixture was diluted with dichloromethane, washed with water, saturated sodium bicarbonate, and brine, dried over sodium bicarbonate, filtered through celite, concentrated, and purified on silica gel to give the product,5-bromo-2-methylfuro[2,3-b]pyridine. Part B:
A mixture of the product, 5-bromo-2-methy[furo[2,3-b]pyridine, from Part A (1 equiv), bispinacolatodiboron (1.2 equiv), PdCi2dppf (0.1 equiv), and potassium acetate (3 equiv) in dioxane was heated at 85 °C until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction was allowed to cool to room temperature, filtered through a pad of celite, concentrated to dryness, treated with diethyl ether, filtered through a pad of celite, and then concentrated to afford the desired product, 2-methyl-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)furo[2,3- b]pyridine.
Synthesis of 5-(4,4.5,5-tetramethyl-1.3i2-dioxaborolan-2-vnthienoF2,3-biDyridine:
Figure imgf000285_0001
SCHEME-9
Part A:
To a solution of 5-bromo-3-((trimethylsiyl)ethynyl)ipyhdin-2-oI (10.63 g, 39.5 mmol) in toluene (195 mL) was added Lawesson's reagent (8.00 g, 19.8 mmol) under nitrogen. The reaction mixture was stirred at 120 °C for 1 hour, and then allowed to cool to 25 °C. Purification by column chromatography (SiO2, 20% ethyl acetate/hexanes) afforded compound as a white solid, 5-bromo-2-(trimethylsilyl)thieno[2,3-b]pyridine, 8.99 g (79%). 1H NMR (300 MHz, CDCI3) δ 8.57 (d, J= 2.2 Hz, 1 H), 8.16 (d, J = 2.2 Hz 1 H), 7.30 (s, 1H), 0.39 (s, 9H).
Part B: The 5-bromo-2-(trimethylsilyl)thieno[2,3-b]pyridine from Part A (8.99 g, 31.4 mmol) was dissolved in ethano! (70 mL), followed by addition of K2CO3 (10.85g, 78.5 mmol). Reaction was stirred at 65 °C for 1 hour. The reaction mixture was allowed to cool to 25 °C, concentrated to dryness, and dissolved in EtOAc (150 mL). The solution was washed with H2O (80 mL), and brine (15OmL), dried over Na3SO4, filtered, and concentrated to yield white solid, 5-bromothieno[2,3-b]pyridine, 6.57 g (98%). 1H NMR (300 MHz, CDCI3) δ 8.60 (d, J = 2.2 Hz, 1 H), 8.16 (d, J = 2.2 Hz 1 H), 7.57 (d, J = 6.0 Hz 1 H), 7.21 (d, J = 6.0 Hz, 1 H). Part C: A mixture of the product, 5-bromothieno[2,3-b]pyridineI from Part B (1 equiv), bispinacolatodiboron (1.2 equiv), PdCI2dppf (0.1 equiv), and potassium acetate (3 equiv) in dioxane was heated at 85 °C until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction was allowed to cool to room temperature, filtered through a pad of celite, concentrated to dryness, treated with diethyl ether, filtered through a pad of celite, and then concentrated to afford the desired product, 5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)thieno[2,3-b]pyridine.
General synthesis of (1 r,4r)-4-(7-amino-3- Aryl,6-substituted pyrazoioM ,5-alPyrimidin- 5-vOcvclohexanecarboxvlic acid:
Figure imgf000286_0001
A mixture of compound {1 r,4r)-tert-butyl 4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yljcyclohexanecarboxylate (1 equiv), boronate (1.75 equiv), palladium catalyst (0.1 equiv), and potassium carbonate (3 equiv) in water and dioxane was stirred at 40 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The mixture was allowed to cool to room temperature, diluted with water and extracted three times with dichloromethane. The combined organics were dried over sodium sulfate, filtered, concentrated, and purified on silica gel to afford compound (1r,4r)-tert- butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyI)amino)-3-aryl-pyrazolo[1 ,5-a]pyrimidin- 5-yi)cyclohexanecarboxylate.
When R = Br:
Compound , (1 r,4r)-tert-butyl 4-(7-(bis((2-{trimethylsilyl)ethoxy)methyl)amino)-3-aryl- pyrazoioJI.5-ajpyrimidin-5-yl)cyclohexanecarboxylate, was stirred in a mixture of dichloromethane, water, and TFA until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The reaction was concentrated and purified on prep-HPLC. The resulting material was stirred with NBS (1.05 equiv) in acetonitrile and DMF at room temperature until the reaction was deemed complete by HPLC1 LC-MS, or TLC analysis. The mixture was concentrated, purified on prep-HPLC, and freeze-dried to afford compound, (1 r,4r)-4-(7-amino-6-bromo-3-aryl - pyrazolo[1 ,5-a]pyrimidin-5- y!)cyclohexanecarboxy!ic acid.
When R = acetyl: Compound, (1 r,4r)-tert-buty! 4-(7-{bis{(2- {trimethylsilyi)ethoxy)methyl)amino)-3-aryl-pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexanecarboxylate , (1 equiv) was stirred with NBS (1.05 equiv) in DMF at room temperature until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The reaction was diluted with ethyl acetate, washed three times with brine, dried over sodium sulfate, filtered, concentrated, and purified on silica gel. The resulting material was stirred with a mixture of vinyl stannane (2 equiv), palladium catalyst (0.1 equiv), and potassium fluoride (3 equiv) in dioxane at 85 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The mixture was allowed to cool to room temperature, diluted with 10% aqueous potassium fluoride, and extracted three times with ethyl acetate. The combined organics were dried over sodium sulfate, filtered, concentrated, and purified on silica gel. The resulting material was stirred with a mixture of HCI in methanol and water at 60 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The mixture was then allowed to cool to room temperature, concentrated, purified by prep-HPLC, and freeze-dried to afford compound (1 r,4r)-4-(6-acetyl-7-amino-3~aryl - pyrazolo[1 ,5-a]pyrimidin-5- yi)cyclohexanecarboxylic acid.
Essentially using the general procedure described above the compounds in the table 4-D can be synthesized.
TABLE-4D
Figure imgf000287_0001
Figure imgf000288_0001
6-Bromo-5-((1-(cvclopropylsulfonyl)pipehdin-4-yl)nnethoxy)-3-(Quinolin-3- yl)pyrazoiof1 ,5-aiPyrimidin-7-amine:
Figure imgf000289_0001
SCHEME- 11
tert-Butv! 4-(f7-(bis(f2-(trinπethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(quinoiin-3- yl)pyrazoio[1 ,5-aiPyrimidin-5-yloxy)methyl)piperidine-1-carboxylate: To a solution of tert-butyl 4-{hydroxymethyl)piperidine-1 -carboxylate (31 1 mg, 1 .44 mmol) and sodium hydride (60% in mineral oil, 58 mg, 1.44 mmol) in THF (2 mf) under argon at room temperature was added 5-chloro-3-(quinolin-3-y!)-N,N-bis((2 (trimethylsilyl)ethoxy)methyl) pyrazolo[1 ,5-a3pyrimidin-7-amine {400 mg, 0.72 mmoi). The mixture was heated at 100°C for 30 min in microwave. Ethyl acetate (50 ml) was added and the mixture was washed with brine and water, dried over Na2SO4, and concentrated in vacuo. The crude product was dissolved in acetonitrile, followed by the addition of N-bromosuccinimide (193 mg, 1.08 mmol). The reaction mixture was stirred at room temperature for 2 h, concentrated in vacuo and purified via silica gel chromatography (10% to 50% ethyl acetate in hexanes gradient) to yield the title compound (0.56 g, 95% yield). LC-MS: 813 [M+H].
6-Bromo-5-(piperidin-4-ylmethoxy)-3-(quinolin-3-yl)pyrazolof1 ,5-a1pyrimidin-7-amine: tert-Butyl 4-({7-(bis((2-(trimethylsi[yl)ethoxy)methyl)amino)-6-bromo-3-(quinolin-3- y[)pyrazolo[1 ,5-a]pyrimidin-5-yloxy)methyl)piperidine-1-carboxylate {0.56 g, 68.9 mmol) was dissolved in methanol (6 ml) and treated with 4N hydrochloride solution in dioxane (4 ml) at 65 °C for 30 min. The reaction solution was concentrated and the crude product (0.48 g) was used in the next step without further purification. LC-MS: 453 [M+H].
6-Bromo-5-((1 -(cvclopropylsulfonyl)piperidin-4-yl)methoxy)-3-(quinolin-3- yi)pyrazolo[1 ,5-alpyrimidin-7-amine:
Cyclopropanesuifonyl chloride {24 mg, 0.167 mmoi) was added into a solution of 6- bromo-5-(piperidin-4~ylmethoxy)-3-(quinolin-3-y!)pyrazoio[1 ,5-a]pyrimidin-7-amine (50 mg, 0.11 1 mmol) and triethylamine (56 mg, 0.553 mmol) in DMF at room temperature. The resulting reaction mixture was stirred at room temperature for 16 h, concentrated and purified by prep-LC to afford the title compound (21.5 mg): LC/MS RT = 4.17 min. Mass calculated for, M+H 557.09, observed 557.09.
Compounds in the table-4-E can be synthesized as described following general procedure dscribed above scheme( 11 )
TABLE-4E
Figure imgf000290_0001
Figure imgf000291_0001
Figure imgf000292_0001
Figure imgf000293_0001
Figure imgf000294_0001
Figure imgf000295_0002
By essentially using the similar experimental conditions used in the scheme 11 , compounds in the table 4F can be synthesized.
TABLE-4F
Figure imgf000295_0001
2-(4-(7-amino-6-bromo-3-(quinolin-3-yl) pyrazolori ,5-aipyrimidin-5-yloxy)phenyl)acetic acid:
Figure imgf000296_0001
SCHEME-12
PartA:
Methyl 2-(4-(7-(bis((2-(trimethyisilyl)ethoxy)methyhamino)pyrazoio[1 ,5-aipyrimiclin-5- yloxy)phenyl)acetate:
The compound methyl 2- (4-hydroxy phenyl) acetate (330 mg, 0.2 mmol) was dissolved in dry DMF (3 mL) and NaH (80 mg, 60% in oil, 0.2 mmol) was added and the mixture was stirred at room temperature for 5 min. Then, 5-chloro-N,N-bis((2-
(trimethylsilyl)ethoxy)methyI)pyrazolo[1,5-a]pyrimidin-7-amine (430 mg, 0.1 mmol) was added and the resulting mixture was heated up to 1300C with microwave and stirred for 30 min. After cooling to room temperature, NH4Cl (aq) was added to quench the reaction and extracted with EtOAc (50 mL x 3). The combined organics were washed with water and brine and dried over Na2SO4. After concentration, the residue was purified with column (silica gel, 20% EtOAc/Hexane) gave the product, methyl 2-(4-(7-(bis((2- (trimethylsiIyi)ethoxy)methyl)amino)pyrazolo[ L,5-a]pyrimidin-5-yIoxy)phenyl)acetate, (490 mg) as clear oil. HPLC-MS tR = 2.77 min (UV254 nm); mass calculated for formula 027H42N4O5Si2 558.3, observed LCMS m/z 559.3 (M+H).
Part B: Methyl 2-(4-(7-(bis((2-(trimethylsiiyl)ethoxy)methyl)amino)-3-iodopyrazoiori ,5- aipyrimidin-5-yloxy)phenvOacetate:
Methyl 2-(4-(7-{bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[l,5-a]pyrimidin-5- yloxy)phenyl)acetate (450 mg, 0.81 mmol) was dissolved in ACN (15 raL) and NIS (200 mg, 0.88 mmol) was added. The resulting mixture was stirred at room temperature for 2 h and then the solvent was removed under reduced pressure. The residue was purified with column (silica gel, 10% EtOAc/Hexane) gave the product, methyl 2-(4-(7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1,5-a]pyrimidin-5-yloxy)phenyl)acetate (543 mg) as yellowish oil. HPLC-MS tR = 2.96 min (UV254 nm); mass calculated for formula C27H4IlN4O5Si2684.2, observed LCMS m/z 685.2 (M+H).
Part C:
Methyl 2-(4-(7-(bisf(2-(trimethylsiiyl)ethoxy)methyπamino)-3-(quinolin-3- yl)pyrazolori ,5-a]pyrSmidin-5-yloxy)phenyl)acetate:
Under Ar, compound, methyl 2-(4-(7-(bis((2-(rrimethylsilyl)ethoxy)methyl)amino)-3- iodopyrazolo[1,5-a]pyrimidin-5-yloxy)phenyI)acetate (180 mg, 0.26 mmol) was mixed with Pd(dppf)Cl2 (21 mg, 0.26 mmol, K3PO4 (106 mg, 0.5 mmol), 3-quinoIine bornic acid (55 mg, 0.31 mmol) and dioxane (10 mL with 1 ml water). The resulting mixture was heated at 90 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (60 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product, methyl 2-(4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3-yl)pyrazolo[l,5-a]pyrimidin-5- yloxy)phenyl)acetate (150 mg). HPLC-MS tR = 2.60 min (UV254 nm); mass calculated for formula C36H47N5O5Si2685.3, observed LCMS m/z 686.3 (M+H).
Part D: methyl 2-(4-(7-(bis((2-(trimethyisilyl)ethoxy)methyl)amino)-6-bromo-3-(quinolin-3- yl)pyrazolori .5-alpyrimidin-5-yloxy)phenyl)acetate:
Compound, methyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinoIin-3- yl)pyrazolo[1,5-a]pyrimidin-5-yloxy)phenyl)acetate (150 mg, 0.22 mmol) was dissolved in ACN (10 mL). NBS (40 mg, 0.22 mmol) was added and the mixture was stirred at room temperature for 1 h. After concentration, the product, methyl 2-(4-(7-(bis((2-
(trimethylsilyl)ethoxyJmethyl)aminoJ-ό-bromo-3-tquinolin-3-yl)pyrazoloCU5-alpyrimidin-5- yloxy)phenyl)acetate was used in the next step without further purification. HPLC-MS tR - 2.70 min (UV254 nm); mass calculated for formula C36H46BrN5O5Si2 763.2, observed LCMS m/z 764.2 (M+H).
Part E: methyl 2-(4-(7-amino-6-bromo-3-(Quinolin-3-yl)pyrazolo[1 ,5-alpyrimidin-5- yloxy)phenvOacetate: The crude methyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyi)amino)-6-bromo-3-(quinolin- 3-yl)pyrazolo[1,5-a]pyrimidin-5-yloxy)pheny[)acetate from above step D was treated with 50% TFAZH2O (3 mL) and stilted at room temperature for 1 h. Solvent was removed to yield thick oil, methyl 2-(4-(7-amino-ό-brorno-3-(quinolin-3-yl)pyrazolo[1,5-a]pyrimidin-5- yloxy)phenyl)acetate, which was used in the next step directly without further purification. HPLC-MS tit = 1.60 min (UV254 nm); Mass calculated for formula C24H18BrN5O3. 503.1, observed LCMS m/z 504.1 (M+ Na).
Part F:
2-(4-(7-amino-6-bromo-3-(quinolin-3-yl) pyrazoloH ,5-a]pyrimidin-5-yloxy)phenyl)acetic acid:
Crude compound , methyl 2-(4-(7-amino-6-brorno-3-(quinolin-3-yl)pyrazolo[1,5-a]pyrimidin- 5 -yloxy) phenyl) acetate from step E was dissolved in THF (5 mL) and LiOH ( IN, 1 mL) was added. The mixture was stirred at room temperature overnight. The solvent was removed under vacuo, and the resulting residue was taken over with water and the pH value was adjusted to 5-6. The solid was collected with filtration and purified with HPLC gave the product, 2-(4-(7- amino-6-bromo-3-(quinolin-3-yl) pyrazolo[1,5-a]pyrimidin-5-yloxy)phenyl)acetic acid. HPLC-MS tR = 1.53 min (UV254 nm); Mass calculated for formula C23Hj6BrN5O3, 489.0, observed LCMS m/z 490.0 (M+H).
By essentially the same procedure given in Preparative Example given above, compounds 4.130-4.144 given in Column 2 of Table 4G can be prepared.
TABLE 4G
Figure imgf000298_0001
Figure imgf000299_0001
Figure imgf000300_0001
Synthesis of 4-(7-amino-6-bromo-3-(6-phenylpyndin-3-yl)pyrazolori ,5-a1pyrimidin-5- yl)benzoic acid: SCHEME- 13
Figure imgf000301_0001
Part A: Syntheisis of methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methy!)amino)pyrazolori ,5- aipyrimidin-5-yl)benzoate:
Under Ar, compound, 5-chloro-N,N-bis((2-(trimemylsilyl)ethoxy)methyl)pyrazolo[l,5- a]pyrirnidin-7-amine, (430 mg, 1.0 mmol) was mixed with Pd(dppf)Cl2 (82 mg, 0.1 mmol, K3PO4 (636 mg, 3.0 mmol), methyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzoate (200 mg, 1.1 mmol) and dioxane (20 mL with 1 mi water). The resulting mixture was heated at 90 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (60 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product, methyl 4-(7-(bis((2- (trimethy!siiyl)ethoxy)methyl)amino)pyrazolo[1,5-a]pyrimidin-5-yi)benzoate (459 mg). HPLC-MS tR = 2.89 min (UV254 πm); mass calculated for formula C26H40N4O4Si2 528.3, observed LCMS m/z 529.2 (M+H).
Part B:
Synthesis of methyl 4-(7-(bis((2-(trimethylsilvt)ethoxy)methyl)amino)-3- iodopyrazolo[1 ,5-a]pyrimidin-5-yl)benzoate:
Compound, methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyI)amino)-3-iodopyrazolo[ 1 ,5- a]pyrimidin-5-yl)benzoate was prepared with the same iodonation condition described in scheme- 12 Part B. HPLC-MS tR = 3.09 min (UV254 ,,m); mass calculated for formula C26H39IN4O4Si2 654.2, observed LCMS m/z 655.2 (M+H).
Part C: Synthesis of methyl 4-(3-(biphenyl-4-yl)-7-(bis((2- (trimethylsilvi)ethoxy)methyl)amino)pyrazolo[1 ,5-alpyrimidin-5-yl)benzoate: Title Compound, methyl 4-(3-(biphenyl-4-yiy7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)pyrazolof 1,5-a1pyrimidin-5-yl)benzoate was prepared with the same coupling condition described in the schem-12 Part C. HPLC-MS IR = 2.98 min (UV254 nm); mass calculated for formula C37H47N5O4S12681.3, observed LCMS m/z 682.2 (M+H).
Part D:
Synthesis of methyl 4-(7-(bisf(2-(trimethylsilyl)ethoxy)methy0amino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-alpyrimidin-5-yl)benzoate:
Title Compound was prepared with the same bromonation condition described in Scheme- 12, Part D . HPLC-MS tR = 3.11 min (UV254 nm); mass calculated for formula C37H46BrN5O4Si2 759.2, observed LCMS m/z 760.2 (M+H).
Part E:
Synthesis of methyl 4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5- aipyrimidin-5-yl)benzoate:
Title compound, methyl 4-(7-amino-6-brorno-3-(6-phenylpyridin-3-yl)pyrazolo[l,5- a]pyrimidin-5-yl)benzoate was prepared with the same deprotection condition described in Scheme- 12 Part E. HPLC-MS tR = 1.88 min (UV254 nm); mass calculated for formula C23H18BrN5O2499.1 , observed LCMS m/z 500.1 (M+H).
Part F: 4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazoio[l,5-a]pyrimidin-5-yl)benzoic acid:
Compound, 4-(7-amino-6-bromo-3-(6-pheny!pyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5- yl)benzoic acid was prepared with the same hydrolysis condition described in Scheme- 12 Part E. HPLC-MS tR = 1.44 min (UV254 nm); mass calculated for formula C24Hi6BrN5O2 485.0, observed LCMS m/z 486.1 (M+H).
By essentially the same procedure given in Preparative scheme-33 above, compound 4.145- 4.166 given in Column 2 of Table 4 H can be prepared from compound 2.
TABLE - 4H
Figure imgf000303_0001
Figure imgf000304_0001
Figure imgf000305_0001
Figure imgf000306_0001
Figure imgf000307_0001
Synthesis of 4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-y0pyrazoloH ,5-a1pyrimidin-5- yl)benzoic acid:
Figure imgf000308_0001
SCHEME-14
Part A methyl 4-(7-(bisff2-(trimethyisilyl)ethoxy)methyl)amino)-6-(1-ethoxyylnyl)-3-(6- phenylpyridin-3-yl)pyrazolori ,5-aipyrimidin-5-yl)benzoate:
Compound, methyl 4-(7-(bis((2-(trimemylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolo[1,5-a]pyrirnidin-5-yl)benzoate (40 mg, 0.053 mmol) under Argon, was mixed with Pd(PPh3)4 (12 mg, 0.01 mmol, (l-ethoxyvinyl)tributylstannane (85 uL, 0.25 mmol) and dioxane (3 mL). The resulting mixture was heated at 100 °C and stirred over night. After cooled to room temperature, the mixture was filtered through 10% KF on silica gel and washed with EtOAc. After concentration, the crude product, methyl 4-(7-(bis((2- (trimethylsily!)ethoxy)methyl)amino)-6-(l-ethoxyvinyl)-3-(6-phenyIpyridin-3-yl)pyrazolo[1,5- a]pyrimidin-5-yl)benzoate was used in the next step directly without further purification. HPLC-MS tR = 3.18 min (UV254 ,™); mass calculated for formula C4[H53N5O5Si2 751.4, observed LCMS m/z 752.3 (M+H).
Part B
Methyl 4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-alpyrimidin-5- yl)benzoate:
Compound, methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-(l-ethoxyvinyl)-3-(6- phenylpyridin-3-yl)pyrazoIo[1,5-a]pyrimidin-5-yl)benzoate was treated with 4N HCi in doioxane for 2hrs to result in the product, methyl 4-(6-acetyl-7-amino-3-(6-phenylpyridin-3- yl)pyrazolo[l,5-a]pyrimidin-5-y[)benzoate purified by prep.HPLC. HPLC-MS tR = 1.85 min (UV254 nm); mass calculated for formula C27H2IN5O3463.2, observed LCMS m/z 464.0 (M+H).
Part C: 4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolori .5-aipyrimidin-5-yl)benzoic acid: Compound methyl 4-(6-acetyf-7-amino-3-(6-phenylpyridin-3-yI)pyrazolo[ l,5-a]pyrimtdin-5- yi)benzoate was treated with 2 equivalents of LiOH and stirred at room temperature for 4hrs. Neutralization with dil.HCl and extraction in to organic layer, gave the product, 4-(6-acetyl-7- amino-3-(6-phenylpyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5-yl)benzoic acid in quantitavive yield. The compound dissolved in to water-acetonitrile and lyophilized to give a white powder. HPLC-MS tR = 1.49 min (UV254 nm); mass calculated for formula C26Hi9N5O3 449.1, observed LCMS m/z 450.2 (M+H).
By essentially the same procedure given in above preparative example , compounds 4.167-4.171 given in Column 2 of Table 41 can be prepared.
TABLE-41
Figure imgf000309_0001
Figure imgf000310_0001
Synthesis of 6-bronno-5-(4-(methylsu[fonyl)phenoxy)-3-(quinolin-3-yl)pyrazolori ,5- aiDyrimidin-7-amine:
Figure imgf000311_0001
SCHEME- 15
Part A:
5-(4-(methylthSo)phenoxy)-N,N-bis((2-(trimethylsilyπethoxy)methyl)pyrazolori ,5- aipyrimidin-7-amine: The 4-(methylthio)phenol (280 mg, 2.0 mmol) was dissolved in dry DMF (3 mL) and NaH (88 mg, 60% in oil, 2.2 mmol) was added and the mixture was stirred at room temperature for 5 min. Then, 5-chloro-N,N-bis((2-(trimemyIsilyl)ethoxy)methyl)pyrazolo[l ,5-a]pyrimidin-7- amine (430 mg, 0.1 mmol) was added and the resulting mixture was heated up to 130 0C with Microwave and stirred for 30 min. After cooling to room temperature, NH4C1 (aq) was added to quench the reaction and extracted with EtOAc (50 mL x 3). The combined organics were washed with water and brine and dried over Na2SO4. After concentration, the residue was purified with column (silica gel, 20% EtOAc/Hexane) gave the product 5-(4- (methylthio)phenoxy)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7- amine (490 mg) HPLC-MS .R = 2,98 min (UV254 nm); mass calculated for formula C25H40N4O3SSi2 532.2, observed LCMS m/z 533.2 (M+H).
Part B:
3-iodo-5-(4-(methyithio)pheπoxy)-N.N-bisff2-
(trimethyisilyl)ethoxy)methyi)pyrazolof1.5-aipyrimidin-7-amine:
5-(4-(methylthio)phenoxy)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5- a]pyrimidin-7-amine (490 mg, 0.81 m) was dissolved in ACN (15 mL) and NIS (200 mg, 0.88 mmol) was added. The resulting mixture was stirred at room temperature for 2 h and then the solvent was removed under reduced pressure. The residue was purified with column (silica gel, 10% EtOAc/Hexane) gave the product, 3-iodo-5-(4-(methylthio)phenoxy)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (541 mg) as yellowish oil.. HPLC-MS tR = 3.18 mln (UV254 nJ; mass calculated for formula C25H39IN4O3SSi2658.1, observed LCMS m/z 659.2 (M+H). Part C:
5-(4-(methylthio)phenoxy)-3-(αuinolin-3-yl)-N.N-bisff2- (trimethylsilvi)ethoxy)methyl)pyrazoiori ,5-aipynmidin-7-amine:
3-iodo-5-(4-(methylthio)phenoxy)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5- a]pyrimidin-7-amine (180 mg, 0.26 mmol) under Argon,was mixed with Pd(dppf)Cl2 (21 mg, 0.26 mmol, K3PO4 (106 mg, 0.5 mmol), 3-quinoline bornic acid (55 mg, 0.31 mmol) and dioxane (10 mL with 1 ml water). The resulting mixture was heated at 90 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (60 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product 5-(4-(methylthio)phenoxy)-3-(quirιolin-3-y I)-N1N- bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (150 mg). HPLC-MS tR = 2.85 min (UV254 nm); mass calculated for formula C34H45N5O3SSi2659.3, observed LCMS m/z 660.1 (M+H). Part D:
5-(4-(methylsuifonyl)phenoxy)-3-(quinoiin-3-yl)-N.N-bis(f2- (trimethy[silyl)ethoxy)methyl)pyrazolori .5-alpyrimidin-7-amine:
Compound, 5-(4-(methylthio)phenoxy)-3-(quinolin-3-yl)-N,N-bis((2- (trimethyIsilyl)ethoxy)methyi)pyrazolo[l,5-a]pyrimidm-7-amine (87 mg, OJ 32 mmol) was dissolved in DCM (5 mL) and m-CPBA (66 mg, -70%, 0.27 mmol) was added. The result mixture was stirred at room temperature for 1 h. NaHCO3 (aq.) was added and the water was extracted with DCM. The combined organics was dried over Na2SO4 and concentrated. The crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product, 5-(4- (methylsulfonyl)phenoxy)-3-(quinolin-3-yl)-N,N-bis((2-
(trimethylsilyI)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine (89 mg). HPLC-MS IR = 2.37 min (UV254 nm); mass calculated for formula C34H4SNsOsSSi2 691.3, observed LCMS m/z 692.2 (M+H). Part E:
6-Bromo-5-(4-(methylsulfonyl)phenoxy)-3-(quinoiin-3-yl)-N,N-bis((2- (tπmethylsiiyl)ethoxy)methyl)pyrazolo[1 ,5-alpyrimidin-7-amine:
Compound, 5-(4-(methylsulfonyl)phenoxy)-3-(quinolin-3-yl)-N,N-bis((2- (trimethyisilyl)ethoxy)methyl)pyrazolo[ 1 ,5-a]pyrimidin-7-amine (150 mg, 0.22 mmol) was dissolved in ACN (10 mL). NBS (40 mg, 0.22 mmol) was added and the mixture was stirred at room temperature for 1 h. After concentration, the residue, 6-bromo-5-(4-
(methylsulfonyl)phenoxy)-3-(quinolin-3-yi)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazoIo[1,5-a]pyrimidin-7-amine was used in the next step without further purification. HPLC-MS IR = 2.66 min (UV254 nm); mass calculated for formula
C34H44BrN5O5SSi2769.2, observed LCMS m/z 770.0 (M+H).
Part F: 6-bromo-5-(4-(methylsu[fonyl)phenoxy)-3-(quinolin-3-yl)pyrazolof1 ,5-alpyrimidin-7- amine:
The crude, 6-bromo-5-(4-(methylsulfonyl)phenoxy)-3-(quinolin-3-yl)-N,N-bis((2- (tritnethylsilyl)ethoxy)methyl)pyrazoIo[1,5-a]pyrimidin-7-amine from above step E was treated with 50% TFA/H2O (3 mL) and stirred at room temperature for 1 h. Solvent was removed to yield thick oil which was purified by prep HPLC to give the final product. 6- bromo-5-(4-(methylsuifonyl)phenoxy)-3-(quinoHn-3-yl)pyrazolo[l,5-a]pyrimidin-7-amine. HPLC-MS tR = 1.37 min (UV254 nm); mass calculated for formula C22Hi6BrN5O3S2509,0, observed LCMS m/z 510.0 (M+H).
By essentially the same procedure given in Preparative above Example, compounds 4.172- 4.174 given in Column 2 of Table 4 J can be prepared.
TABLE 4-J
Figure imgf000313_0001
Figure imgf000314_0002
Synthesis of 2-(4-((7-amino-6-bromo-3-(quinolin-3-yltoyrazolori ,5-alpyrimidin-5- yl)methyDphenvOacetic acid :
Figure imgf000314_0001
SCHEME- 16
Part A:
Methyl 2-(4-((4,4,5,5-tetramethyl-1.3.2-dioxaborolan-2-yl)methyπphenyl)acetate:
Methyl 2-(4-(bromomethyl)phenyl)acetate (1.03 g, 4.24 mmol) was mixed with Pd(PPh3)4 (500 mg, 0.4 mmol, 4,4,41,4',5,5,51,5'-octamethyl-2,21-bi(l,3,2-dioxaborolane) (1.27 g, 5 mmol), K2CO3 (1.75 g, 12.7 mmol) and dioxane (20 mL) under inert Argon atmoshphere. The resulting mixture was heated at 80 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (100 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 10~30% EA/hexane) gave the product, methyl 2-(4-((4,4,5,5-tetramethyl- [,3,2-dioxaborolan-2-yl)methyl)phenyl)acetate (721 mg) as semi- solid. HPLC-MS tR = 2.17 min (UV254 πm); mass calculated for formula Ci6H23BO4290.2, observed LCMS m/z 291.3 (M+H).
Part B:
Methyl 2-(4-((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a1pyπmidin-5- yl)methyDphenvOacetate:
Under Ar, 5-chloro-N,N-bis((2-(trimethylsiIyl)ethoxy)methyl)pyrazolo[1,5-a]pyrirnidin-7- amine (429 mg, 1.0 mmol) was mixed with Pd(dppf)C12 (82 mg, 0.1 mmol, boronate, methyl 2-(4-((4,4,5,5-tetramethyl-I,3,2-dioxaborolan-2-yl)methyl)phenyl)acetate (4go mg, 1.65 mmol, not very pure), K3PO4 (424 mg, 2.0 mmol) and dioxane (10 mL). The resulting mixture was heated at 100 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (100 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 10-30% EA/hexane) gave the product, methyl 2-(4-((7- (bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1,5-a]pyrimidin-5- y!)methyl)phenyl)acetate (188 mg) and recovered the starting material 5-chloro-N,N-bis((2- (trimethylsilyI)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (289 mg). HPLC-MS IR = 2.50 min (UV254 nm); mass calculated for formula C28H44N4O4Si2 556.3, observed LCMS m/z 557.3 (M+H).
Part C:
Methyl 2-(4-((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5- a1pyrimidin-5-yl)methyl)phenyl)acetate: methyl 2-(4-((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[ 1 ,5-a]pyrimidin-5- yl)methyl)phenyl)acetate (450 mg, 0.81 mmol) was dissolved in ACN (15 mL) and NIS (200 mg, 0.88 mmol) was added. The resulting mixture was stirred at room temperature for 2 h and then the solvent was removed under reduced pressure. The residue was purified with column (silica gel, 10% EtOAc/Hexane) gave the product methyl 2-(4-((7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1,5-a]pyrimidin-5- yi)methyl)phenyi)acetate (541 mg) as yellowish oil. HPLC-MS .R = 2.97 min (UV2g4 nm); mass calculated for formula C28H43IN4O4Si2682.2, observed LCMS m/z 683.2 (M+H). Part D:
Methyl 2-(4-(f7-(bis(f2-(trimethylsilyl)βthoxy)methyl)amino)-3-(quinolin-3- yl)pyrazoloH ,5-a1pyrimidin-5-yl)methyl)phenyl)acetate:
Under Ar, methyl 2-(4-((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1,5- a]pyrimidin-5-yl)methyl)phenyl)acetate (180 mg, 0.26 mmol) was mixed with Pd(dppf)Cl2 (21 mg, 0.26 mmol, K3PO4 (106 mg, 0.5 mmol), 3-quinoline bornic acid (55 mg, 0.31 mmol) and dioxane (10 mL with 1 ml water). The resulting mixture was heated at 90 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (60 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product methyl 2-(4-((7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3-yl)pyrazolo[l ,5-a]pyrimidin-5- yl)methyl)phenyl)acetate (150 mg).HPLC-MS tR = 2.50 min (UV254 nm); mass calculated for formula C37H49N5O4Si2683.3, observed LCMS m/z 684.4 (M+H). Part E:
Methyl 2-(4-((7-(bis((2-(thmethylsilyl)ethoxy)methyl)amino)-6-bromQ-3-(quinolin-3- vQpyrazoloH .5-aipyrimidin-5-yl)methyl)phenyl)acetate:
Methyl 2-(4-{(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quLnolin-3-yl)pyrazolo[l,5- a]pyrimidin-5-yI)methyl)phenyl)acetate (150 mg, 0.22 mmol) was dissolved in ACN (10 mL).
NBS (40 mg, 0.22 mmol) was added and the mixture was stirred at room temperature for 1 h, After concentration, the product that resulted, methyl 2-(4-((7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(quinolin-3-yl)pyrazoio[1,5-a]pyrimidin-5- yl)methyl)phenyl)acetate was used in the next step without further purification
HPLC-MS tR = 2.66 min (UV254 nm); mass calculated for formula C37H48BrN5O4Si2 761.2, observed LCMS m/z 762.2 (M+H).
Part F:
Methyl 2-(4-(f7-amino-6-bromo-3-(αuinolin-3-yl)pyrazoloπ .5-a1pyrimidin-5- yl)methyDphenypacetate: The crude methyl 2-(4-((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(quinolin- 3-yI)pyrazolo[1,5-a]pyrimidin-5-yl)methyl)phenyl)acetate from above step E was treated with 50% TFA/H2O (3 mL) and stirred at room temperature for 1 h. Solvent was removed to yield thick oil, Methyl 2-(4-((7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[l,5-a]pyrimidin-5- yl)methyl)phenyl)acetate which was used in the next step directly without further purification. HPLC-MS tR = 1.72 m in (UV254 nm); mass calculated for formula C25H20BrN5O2 501.1, observed LCMS m/z 502.0(M+H).
Part G:
2-(4-((7-amino-6-bromo-3-(quinolin-3-yl)pyrazoio[1 T5-alpyrimidin-5- yl)methyl)phenyl)acetic acid :
Crude Methyl 2-(4-((7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1,5-a]pyrimidin-5- yl)methy[)phenyl)acetate from step F was dissolved in THF (5 mL) and LiOH (IN, i mL) was added. The mixture was stirred at room temperature overnight. The solvent was removed under vacuo, and the resulting residue was taken over with water and the pH value was adjusted to 5-6. The solid was collected with filtration and purified with HPLC gave the product 2-(4-((7- amino-ό-bromo-3-tquinolin-3-ylJpyrazolofU5-alpyrimidin-5-ylJmethyl)phenylJacetic acid. HPLC-MS CR = 1.53 min (UV254 nm); mass calculated for formula C24Hi8BrN5O2487.1, observed LCMS m/z 487.9 (M+H).
By essentially same procedure given, in preparative above example, compounds 4.175 & 4. 176 given in Column 2 of Table 4Kcan be prepared.
TABLE-4 K
Figure imgf000317_0002
Synthesis of 6-bromo-5-(4-(methylsulfonyl)benzvn"3-(quinoiin-3-yl)pyrazolof1 ,5- alpyrimidin-7-amine:
Figure imgf000317_0001
SCHEME-17 Essentially the steps from A through G described in the example, synthesis of 2-(4-((7-amino- 6-bromo-3-(quinolin-3-yl)pyrazolo[J,5-a]pyrimidin-5~yi)methyl)phenyI)acetic acid can be used for this synthesis of 6-bromo-5-(4-(methylsulfonyI)benzyl)-3-(quinolin-3-yl)pyrazolo[1,5- a] py rim id in-7 -am ine
Part A:
4,4,5, 5-tetramethyi-2-(4-(methylthio)benzyl)-1 ,3,2-dioxaboroiane: 4,4,5, 5-tetramethyl-2-(4-(methyIthio)benzyl)- [,3,2-dioxaborolane prepared with the same condition described in above example Part A. HPLC-MS _R = 2.23 min (UV254 nm); mass calculated for formula Ci4H2IBO2S 264.1, observed LCMS m/z 265.2 (M+H).
Part B: 5-(4-(methylthio)benzyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5- aipyrimidin-7-amine:
Compound, 5-(4-(methylthio)benzyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[ 1 ,5- a]pyrimidin-7-amine was prepared with the same coupling condition described in above example Part B. HPLC-MS ta = 2.74 min (UV254 nm); mass calculated for formula C26H42N4O2SSi2530.3, observed LCMS m/z 531.2 (M+H).
Part C:
5-(4-(methylsulfonyl)benzyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5- aipyrimidin-7-amine:
Compound, 5-(4-(methylthio)benzyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[ 1,5- a]pyrimidin-7-amine (87 mg, 0.132 mmol) was dissolved in DCM (5 niL) and m-CPBA (66 mg, -70%, 0.27 mmoi) was added. The result mixture was stirred at room temperature for 1 h. NaHCO3 (aq.) was added and the water was extracted with DCM. The combined organics was dried over Na2SO4 and concentrated. The crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product 5-(4-(methy[sulfonyl)benzyl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (89 mg). HPLC-MS tR = 2.52 min (UV2S4 nm); mass calculated for formula C26H42N4O4SSi2 562.2, observed LCMS m/z 563.3 (M+H).
Part D:
3-iodo-5-(4-(methylsulfonvhbenzyl)-N.N-bisff2-
(trimethyisilyl)ethoxy)methyl)pyrazolof1 ,5-aipyrimidin-7-anπine:
Compound, 3-iodo-5-(4-(methylsulfonyl) benzyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl) pyrazoio [1,5-a]pyrimidin-7-amine was prepared with the same iodonation condition described in above example part C. HPLC-MS tR = 2.81 min (UV254 nm); mass calculated for formula C26H4IlN4O4SSi2 688.1, observed LCMS m/z 689.2 (M+H).
Part E:
5-(4-(methylsulfonyl)benzvn-3-(αuinoiin-3-yl)-N,N-bisf(2- ftrimethvlsilvπethoxv)methvl)pvrazolof1.5-alpvrimidin-7-amine: Compound, 5-(4-(methylsulfonyl)benzyl)-3-(quinolin-3-yϊ)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoio[1,5-a]pyrimidin-7-arnine was prepared with the same coupling condition described in the above example Part D. HPLC-MS tR = 2.35 mm (UV2S4 πm); mass calculated for formula C35H47N5O4SSi2689.3, observed LCMS m/z 690.3 (M+H).
Part F: β-bromo-5-(4-(methylsulfonyl)benzyl)-3-(quinoiin-3-yl)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazoiori ,5-alpyrimidin-7-amine:
Compound, 6-bromo-5-(4-(methylsulfonyl)benzyl)-3-(quinoIin-3-yI)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[l ,5-a]pyrimidin-7-amine was prepared with the same bromonation condition described in the above example 1 Part E. HPLC-MS IR = 2.52 min (UV254 nm); mass calculated for formula C35H46BrN5O4SSi2 767.2, observed LCMS m/z 768.2 (M+H).
Part G:
6-bromo-5-(4-(methylsulfonyl)benzyl)-3-(quinolin-3-yl)pyrazolof1 ,5-alpyrimidin-7- amine:
Ttile compound, 6-bromo-5-(4-(methylsulfonyl)benzyl)-3-(quinolin-3-yl)pyrazolo[1,5- a]pyrimidin-7-amine was prepared with the same deprotection condition described in example I Part F. HPLC-MS tR = 1.49 min (UV254 nm); mass calculated for formula C23Hj8BrN5O2S 507.0, observed LCMS m/z 5O8.O(M+H).
By essentially the same procedure given in preparative above example, compound 4.177-4.178 given in Column 2 of Table 4L can be prepared.
TABLE 4L
Figure imgf000319_0001
Figure imgf000320_0002
Synthesis of 4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5- yloxy)benzoic acid:
Figure imgf000320_0001
SCHEME-18 Part A:
Methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolof1 ,5-a1pyrimidin-5- yloxy)benzoate:
The methyl 4-hydroxybenzoate (330 mg, 0.2 mmol) was dissolved in dry DMF (3 niL) and NaH (8 mg, 60% in oil, 0.2 mmol) was added and the mixture was stirred at room temperature for 5 min. Then, 5-chloro-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin- 7-amine (430 mg, 0.1 mmol) was added and the resulting mixture was heated up to 130 oC with Microwave and stirred for 30 min. After cooling Co room temperature, NH4CI (aq) was added to quench the reaction and extracted with EtOAc (50 mL x 3). The combined organics were washed with water and brine and dried over Na2SCW. After concentration, the residue was purified with column (silica gel, 20% EtOAc/Hexane) gave the product, methyl 4-(7-(bis((2- (trimemylsilyl)ethoxy)methyl)amino)pyrazolo[1,5-a]pyrimidin-5-yloxy)benzoate (490 mg) as clear oil. HPLC-MS IR = 2.84 min (UV254 πm); mass calculated for formula C26H4ON4O5Si2 544.3, observed LCMS m/z 545.3 (M+H).
Part B:
Methyl 4-(7-{bisff2-(trimethylsiiyl)ethoxy)methynamino)-3-iodopyrazolof1 ,5- a1pyrimidin-5-yloxy)benzoate:
Methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[ 1 ,5-a]pyrimidin-5- yioxy)benzoate (450 mg, 0.81 mmol) was dissolved in ACN ( 15 mL) and NIS (200 mg, 0.88 mmol) was added. The resulting mixture was stirred at room temperature for 2 h and then the solvent was removed under reduced pressure. The residue was purified with column (silica gel, 10% EtOAc/Hexane) gave the product, methyl 4-(7-(bis((2-
(trimethylsilyl)ethoxy)methyl)ammo)-3-iodopyrazolo[1,5-a]pyrimidin-5-yloxy)benzoate (541 mg) as yellowish oil. HPLC-MS CR = 3.10 min (UV254 nm); mass calculated for formula C26H39IN4O5Si2 670.2, observed LCMS m/z 671.2 (M+H). Part C:
Methyl 4-(7-(bis(f2-(trimethylsiiyl)ethoxy)methyπamSnoV3-(6-phenylpyridin-3- v])pyrazoiof1 ,5-a1pyrimidin-5-yloxy)benzoate:
Under Ar, methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1,5- a]pyrimidin-5-yloxy)benzoate (180 mg, 0.26 mmol) was mixed with Pd(dppf)Cl2 (21 mg, 0.26 mmol, K3PO4 (106 mg, 0.5 mmol), 3-quinoline bornic acid (55 mg, 0.31 mmol) and dtoxane (10 mL with 1 ml water). The resulting mixture was heated at 90 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (60 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3- (6-phenyIpyridin-3-yl)pyrazoIo[1,5-a]pyrimidin-5-yIoxy)benzoate. HPLC-MS tR = 2.77 min (UV254 nm); mass calculated for formula C37H47N5O5Si2697.3, observed LCMS m/z 698.2 (M+H). Part D:
Methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenylpyridin-3- yl)pyrazolori ,5-a1pyrimidin~5-yloxy)benzoate:
Compound, methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3- yl)pyrazolo[1,5-a]pyrimidin-5-yloxy)benzoate (150 mg, 0.22 mmol) was dissolved in ACN (10 mL). NBS (40 mg, 0.22 mmol) was added and the mixture was stirred at room temperature for
1 h. After concentration, the residue, methyl 4-(7-(bis((2-
(trimethylsiIyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[l,5- alpyrimidin-5-yloxy)benzoate was used in the next step without further purification. HPLC-MS tR = 3.06 min (UV254 πm); mass calculated for formula C37H46BrN5O5Si2775.2, observed LCMS m/z 776.0 (M+H).
Part E: Methyl 4-(7-(bisf(2-(trimethylsiiyl)ethoxy)methyl)amino)-6-(1 -ethoxyylnyl)-3-(6- phenylpyridin-3-yl)pyrazolof1 ,5-aipyrimidin-5-yloxy)benzoate:
Under Ar, methyl 4-(7-(bis((2-(trimethylsiIyl)ethoxy)methyl)amino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5-yIoxy)benzoate (180 mg, 0.26 mmoi) was mixed with Pd(dppf)Cl2 (21 mg, 0.26 mmol, K3PO4 (106 mg, 0.5 mmol), 3-phenyl pyridyl bornic acid (75 mg, 0.31 mmol) and dioxane (10 mL with 1 ml water). The resulting mixture was heated at 90 °C and stirred over night. After cooled to room temperature, the mixture was diluted with EtOAc (60 mL) and filtered through celite. After concentration, the crude was purified with column (silica gel, 0-30% EtOAc/Hexane) gave the product 5 (150 mg). which was used in the next step directly without further purification. HPLC-MS tβ = 2.96 min (UV254 πm); mass calculated for formula C4IH53N5O6Si2 767.4, observed LCMS m/z 768.4 (M+H).
Part F:
Methyl 4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5- yloxy)benzoate :
The crude from above step E was treated with 50% TFA/H2O (3 mL) and stirred at room temperature for 1 h. Solvent was removed to yield thick oil, methyl 4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-6-(l-ethoxyvinyl)-3-(6-phenylpyridin-3-yl)pyrazolo[1,5- a]pyrimidin-5-yioxy)benzoate. HPLC-MS IR = 1.84 min (UV254 nm); mass calculated for formula C27H21N5O4 479.2, observed LCMS m/z 480.2 (M+H).
Part G:
4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazoiof1 ,5-a]pyrimidin-5-yloxy)benzoic acid;
Crude from step F was dissolved in THF (5 mL) and LiOH (IN, 1 mL) was added. The mixture was stirred at room temperature overnight. The solvent was removed under vacuo, and the resulting residue was taken over with water and the pH value was adjusted to 5-6. The solid was collected with filtration and purified with HPLC gave the product 4-(ό-acetyl-7- amino-3-(6-phenylpyridin-3-yl) pyrazolo[1,5-a]pyrimidin-5-yloxy)benzoic acid.HPLC-MS IR = 1.49 min (UV254 πm); mass calculated for formula C26H19N5O4465.1, observed LCMS m/z 466.2 (M+H).
By essentially the same procedure given in preparative above example(scheme-18), compound 4.179 given in Column 2 of Table 4M can be prepared.
TABLE 4M
Figure imgf000322_0001
Figure imgf000323_0002
Synthesis of 4-(7-amino-6-cvano-3-(6-phenylpyridin-3-yl)pyrazolori ,5-aipyrimidin-5- yl)benzoic acid:
Figure imgf000323_0001
SCHEME-19
Part A
Methyl 4-(7-(bis((2-(trimethylsilyi)ethoxy)methyl)amino)-6-cyano-3-(6-phenylpyridin-3- yl)pyrazolo[1,5-a]pyrimidin-5-yl)benzoate:
Under Ar, compound, methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolo[3 ,5-a]pyrimidin-5-yi)benzoate (80 mg, 0.1 mmol) was mixed with Pd(PPh3)4 (12 mg, 0.01 mmol, Pd(t-Bu3P)3 (5 mg, 0.01 mmol), cyanotributylstannane (85 uL, 0.3 mmol) and dioxane (3 mL) in sealed tube. The resulting mixture was heated at 160 °C and stirred 2 hrs. After cooled to room temperature, the mixture was filtered through 10% KF on silica gel and washed with EtOAc. After concentration, the crude was used in the next step directly without further purification. HPLC-MS tR = 3.18 min (UV254 nm); mass calculated for formula C38H46N6O4Si2 706.3, observed LCMS rπ/z 707.2 (M+H). Part B
Methyl Φfy-amino-β-cvano-3-(6-phenylpyriclin-3-yl)pyrazolori .5-aipyπmidin-5- yQbenzoate: The crude from above step A was treated with 50% TFA/H2O (3 niL) and stirred at room temperature for 1 h. Solvent was removed to yield thick oil which was used in the next step directly without further purification. HPLC-MS t.R = 2.15 min (UV254 nm); mass calculated for formula C26Hi8N6O2446.1, observed LCMS m/z 447.1 (M+H). Part C:
4-(7-amino-6-cvano-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yi)benzoic acid:
Crude compound from step B was dissolved in THF (5 mL) and LiOH (IN, 1 mL) was added. The mixture was stirred at room temperature overnight. The solvent was removed under vacuo, and the resulting residue was taken over with water and the pH value was adjusted to 5~6. The solid was collected with filtration and purified with HPLC gave the product 4-(7-amino-6- cyano-3-(6-phenylpyridin-3-yl) pyrazolo[1,5-a]pyrimidin-5-yl)benzoic acid. HPLC-MS tR - 1.40 min (UV254 nm); mass calculated for formula C25Hi6NeO2432.1, observed LCMS m/z 433.1 (MH-H).
Figure imgf000324_0001
SCHEME-20
Synthesis of Ethyl 1 ,4-dioxaspiror4.51decane-8-carboxylate:
Figure imgf000325_0001
Ethyl 4-oxocyclohexane carboxylate (122.56 mmol, 20.86 g) and para-toluenesulfonic acid monohydrate (12.56 mmol, 2.33 g) are charged to a 1000 rriL roundbottom flask. To this flask was added benzene (300 mL) followed by ethylene glycol (0.37 mol, 20.5 mL). The resulting bi-layer reaction mixture was refluxed overnight in a Dean-Stark trap. Upon cooling, the reaction mixture was diluted in 300 mL DCM and washed with 300 mL saturated NaHCO3(aq)- The acqueous layer was washed with DCM (3X) and the combined organics are dried over Na2SO4. The residue was disolved in DCM and purified by silica gel chromatography on 0% to 50% ethyl acetate in hexanes gradient. The product is tracked via PMA stain. Chromatography yields 17.05 grams (79.58 mmol, 65% yield) ethyl 1 ,4-dioxaspiro[4.5]decane-8-carboxylate.
Synthesis of 1 ,4-DioxasDiro[4.51decane-8-carboxylic acid:
Figure imgf000325_0002
Ethyl 1 ,4-dioxaspiro[4.5]decane-8-carboxylate (79.58 mmol, 17.05 g) is taken up in 300 mL 2:1 THF:H2O in 500 mL roundbottom flask. Lithium hydroxide monohydrate (120 mmoi, 5.01 g) and 100 mL MeOH were added to this solution. The reaction mixture was allowed to stir overnight at room temperature. The reaction mixture was acidified to pH 3 with 1 N HCI(aq) and extracted with 100 mL DCM (x5). The combined organics were combined and then dried over Na2SO4, and the soivent was removed in vacuoto yield 1 ,4-dioxaspiroE4-5]decane-8-carboxyiic acid (14.96 g, 100% yield) as white solid.
Synthesis of Ethyl 3-oxo-3-(1 ,4-dioxaspirof4.51decan-8-yl)propanoate:
Figure imgf000326_0001
1 ,4-Dioxaspiro[4.5]decane-8-carboxylic acid (80.34 mmol, 14.96 g) is charged to a 500 mL roundbottom flask. To this flask is added anhydrous THF (200 ml_), followed by N, N'-carbonyldiimidazole (96.41 mmol, 15.63 g). After vigorous release of CO2 gas, the solution was flushed with argon, sealed, and allowed to stir overnight at room temperature under argon.
After 18 hours, in a separate, sealed and argon-flushed 1000 mL roundbottom flask, LiHMDS (1.0 M in THF1 168.7 mmol) is added to 200 mL anhydrous THF stirring at -78° C. To this solution is added dropwise anhydrous ethyl acetate (173 mmoi, 16.9 mL). This solution is allowed to stir at -78° C for 1 hour prior to dropwise addition of original CDi/acid solution that has been stirred overnight. The reaction mixture was allowed to stir and warm to room temperature overnight.
The reaction was then quenched with saturated NH4CI(aq) (500 mL) and extracted with Et2O (x2). The combined organics are then washed with H2O, saturated brine, and dried over Na2SO4. The solvent is removed in vacuo and the residue is taken up in DCM. The reaction mixture was purified with silica gel chromatography (0% to 60% ethyl acetate in hexanes gradient) to yield ethyl 3-oxo-3- (1 ,4-dioxaspiro[4.5]decan-8-y!)propanoate (1 1.26 g, 55% yield) as pale yellow oil.
Synthesis of 5-( 1 ,4-dioxaspiror4.51decan-8-yl)pyrazolo[1 ,5-aipyrimidin-7-ol:
Figure imgf000327_0001
To a 20 ml_ scintillation vial containing ethyl 3-oxo-3-(1,4-dioxaspiro[4.5]decan-8- yl)propanoate {1 1.7 mmol, 3.00 g) is added 3-amino-1 H-pyrazole (11.7 mmol, 973 mg). The two oils were mixed and heated neat at 100° C for 3 hours. The resulting off-white solid was dissolved up in EtOH (100 ml_) and reduced in vacuo to remove water formed during cyclization. This solid was used in the next step without further purification.
Synthesis of 7-chloro-5-(1 ,4-dioxaspiro[4.51decan-8-yl)pyrazolo[1 ,5-aipyrimidine:
Figure imgf000327_0002
5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[1 ,5-a]pyrimidin-7-ol (11.7 mmol) is charged to 250 ml_ roundbottom flask. To this fiask is added 4 mL N, N-dimethylaniline, followed by 40 mL POCl3. This suspension was sonicated to break up the starting material and stirred at room temperature for 18 hours. After 18 hours, a solution formed. The solution was reduced in vacuo and cooled to 0° C in ice bath. The reaction is then quenched with sat. NaHCO3(aq) and extracted with DCM (x3). The combined organics are dried with Na2SO4 and the solvent was removed in vacuo. The resulting oil was purified via silica gel column on 20% to 100% ethyl acetate in hexanes gradient to yield 7-chloro-5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazoio[1 ,5- a]pyrimidine (2.65 grams, 77% across 2 steps) as white solid.
Synthesis of 5-( 1 ,4-dioxaspirof4.51decan-8-yi)pyrazolof 1 ,5-alpyrimidin-7-amine:
Figure imgf000328_0001
7-chloro-5-{1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[1 ,5-a]pyrimidine (5.58 mmol, 1.64 g) is charged to 10-20 ml microwave vessel. To this vessel is added 10 mL ~7N NH3 in methanol. The vessel was sealed and heated at 100° C for 18 hours. After 18 hours, the reaction mixture was cooled to room temperature and diluted with 100 mL DCM. The resulting solution is washed with saturated NaHCO3(aq) and extracted with DCM twice more. The combined organics were dried over Na2SO4 and the solvent removed in vacuo to yield 5-{1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazoto[1 ,5-a]pyrimidin-7- amine (1.52 g, 99% yield) as pale orange solid.
Synthesis of 4-(7-aminopyrazolod ^-alpyrimidin-5-vPcvclohexanone:
Figure imgf000328_0002
In a 40 mL scintillation vial is combined 5 (1 ,4 dioxaspiro[4.5]decan-8-yl)pyrazoio[1,5- a]pyrimidin-7-amine {5.54 mmol, 1.52 g), EtOH (10 mL), H2O (4 mL) and 4N
HCkdioxane (4 mL). The vial was capped, sealed and the reaction was heated to 80° C overnight. After 18 hours, the reaction mixture was cooled to room temperature and diluted with DCM (100 mL). The solution was washed with saturated NaHCO3^ and extracted with DCM twice more. The combined organics are dried over Na2SO4 and the solvent removed in vacuo to yield 4-(7-aminopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexanone (1.18 g, 93% yield) as pale orange solid.
Synthesis of 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolori ,5-aipyrimidin- 5-yl)cyclohexanone:
Figure imgf000329_0001
4-(7-aminopyrazolo[1 ,5-a]pyrimidin-5-y!)cyclohexanone (5.12 mmol, 1.18 g) was dissolved in DCM (15 mL). To this solution was added N, NJ-diisopropyiethylamine (17.94 mmol, 3.13 mL). The resulting solution was stirred at room temperature while 2-(trimethylsilyl)ethoxymethyl chloride (SEM-CI, 17.94 mL, 3.17 mL) in DCM (5 mL) was added dropwise. After the addition was completed, the reaction mixture was stirred at 50° C for 10 minutes. The solvent was removed in vacuo, and the residue was purified on silica gel column (0% to 60% ethyl acetate in hexanes gradient) to yield 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexanone (1.87 g, 74% yield) as pale yellow oil.
Synthesis of Ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a1pyrimidin-5-yl)cvclohexylidene)acetate:
Figure imgf000329_0002
A 20 mL scintillation vial containing 5 mL THF was charged sodium hydride (60% w/w in mineral oil, 448 μmol, 18 mg). This suspension was broken up via sonication. Triethyl phosphonoacetate (448 μmol, 89.3 μL) in THF (2 mL) was added dropwise. The resulting solution was stirred at room temperature for 10 minutes. To this solution was slowly added a solution of 4-(7-(bis((2-
(trimethy!silyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyr!midin-5-yl)cyclohexanone (408 μmol, 200 mg) in THF (2 mL). The reaction mixture was stirred for 18 hours at room temperature. After 18 hours, the reaction mixture was diluted with DCM (25 mL) and washed with H2O. The aqueous phase was washed with DCM (x2), and the combined organics was dried over NaaSOtt. The solvent was removed in vacuo to yield ethyl 2-(4-(7-(b!s((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin- 5-yl)cyclohexyJidene)acetate (222 mg, 97% yield) as pale yellow oil.
Synthesis of 2-(4-(7-(Bis((2-(trimethy]silvOethoxy)methyhamino)pyrazolori .5- alpyrimidin-5-yl)cvclohexylidenetecetonitriie:
Figure imgf000330_0001
2-(4-(7-{Bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexy!idene)acetonitrile was synthesized in a manner similar to the synthesis of ethyl 2-{4-(7-(bis((2-{trimethy!silyi)ethoxy}nnethyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyiidene)acetate, but with diethyl cyanomethylphosphonate substituted for triethyl phosphonoacetate.
Synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsiiyl)ethoxy)methyl)amino)pyrazolo[1 ,5- aipyrimidin-5-yi)cvclohexyl)acetate:
Figure imgf000330_0002
To a 50 ml_ roundbottom flask was charged ethyl 2~(4-(7-(bis((2-(trimethylsily!)ethoxy)- methyl)amino)pyrazolo[1.5-alpyπmidin-5-yl)cyclohexylideneJacetate (800 mg, 1.43 mmol) and ethyl acetate (15 ml_). The flask was flushed with argon, and 5% palladium on carbon (100 mg) was added. The flask was sealed and degassed under vacuum. Hydrogen gas was then added via balloon. The reaction was stirred under a hydrogen atmosphere for 18 hours. The reaction was then filtered through celite to yield ethyl 2-(4-(7-(bis((2-(trimethylsi!yl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin- 5-y!)cyclohexyl)acetate (761 mg, 1.35 mmol, 95% yield) as pale yellow oii.
Synthesis of 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy^methyl)amino)pyrazolori ,5- alpyrimidin-5-vOcvclohexyl)acetonitriie:
Figure imgf000331_0001
2-(4-(7-(Bis((2-{trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetonitriie was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-{bis((2-(trimethylsiIyl)ethoxy)methyl)amino)pyrazolo[1 ]5-a]pyrimidin-5- yl)cyclohexyl)acetate, but with 2-(4-{7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyc!ohexyiidene)acetonitrile substituted for ethyl 2-(4~(7-(bis((2- (trimethylsityl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexylidene)acetate.
Synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsilvhethoxyϊmethyπamino)-3- iodopyrazoloπ ^-aipyrimidin-5-yl)cvclohexyl)acetate:
Figure imgf000331_0002
A 50 mL roundbottom flask was charged ethyl 2-(4-(7~{bis((2-(trimethylsilyl)ethoxy)- methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5-y!)cyclohexyl)acetate (381 mg, 0.68 mmol) and acetonitrile (10 mL). To this soiution was added N-lodosuccinimtde (167 mg, 0.74 mmol). The reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo and the crude oil was purified via silica gel chromatography (0% to 30% ethyl acetate in hexanes gradient) to yield ethyl 2-(4-(7- (bis((2-(trimethylsily!)ethoxy)methy!)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate (335 mg, 0.49 mmol, 72% yield) as clear oil.
Synthesis of 2-(4-(7-(bis((2-(tπmethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolof 1 ,5- alpyrimidin-3-yl)cvclohexyl)acetonitrile:
Figure imgf000332_0001
2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 !5-a]pyrimidin-5- yl)cyclohexyi)acetonitrile was synthesized in a manner similar to the synthesis of ethyl
2-(4-(7-(bis((2-(trimethyIsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate, but with 2-(4-(7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetonitrile substituted for ethyl 2-(4-(7-(bis((2- (trimethylsilyl)ethoxyJmethylJaminoJpyrazoloJi .5-alpyrimidin-5-yl)cyclohexyl)acetate.
Synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsilvnethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazoloπ .5-aipyrimidin-5-yl)cvclohexyl)acetate:
Figure imgf000333_0001
To a 40 ml_ scintillation via! was charged 3-quinoline boronic acid (0.73 mmol, 127 mg), K3PO4 (1.46 mmol, 310 mg) and PdC!2(dppf) • CH2CI2 (.049 mmol, 40 mg). To this mixture was added a solution of ethyl 2-(4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1,5-a]pyrimidin-5- yl)cyciohexyl)acetate (335 mg, 0.49 mmol) in dioxane (9 m!_). To this suspension was added distilled H2O (1 m!_). The resulting solution was stirred at 100° C for 18 hours. The reaction was concentrated in vacuo and then purified via silica gel chromatography (0% to 60% ethyl acetate in hexanes gradient) to yield ethyl 2-{4-(7- (bis((2-(trimethylsiiyl)ethoxy)methyl)amino)-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate (198 mg, 287 μmol, 59% yield) as yellow oil.
Synthesis of 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyπamino)-3-(αuinolin-3- yl)pyrazoloH ^-alpyrimidin-5-vOcvclohexyl)acetonitrile:
Figure imgf000333_0002
2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyI)amrno)-3-(quinolin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)acetonitrile was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis((2-(thmethylsilyi)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-y!)cyclohexyl)acetate, but with 2-(4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetonitrile substituted for ethyl 2-(4-(7-(bis((2- (trimethylsily[)ethoxy)methy!)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate. Synthesis of ethyl 2-(4-(7-(bis((2-(trimethyisilvBethoxy)methyπamino)-6-bromo-3- (quinolin-3-yl)pyrazolori ,5-aipyrirnidin-5-yltovclohexyl)acetate:
Figure imgf000334_0001
To a 25 mL roundbottom flask was charged ethyl 2-(4-(7-{bis((2-(trimethylsi!yl)ethoxy)- methyl)amιno)-3-(quinolin-3-yl}pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetate (198 mg, 287 μmoi) and acetonitrile (10 mL). To this solution was added N- bromosuccinimide (56 mg, 316 μmol). The resulting reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo and the resulting oil was then purified via silica gel chromatography (0% to 30% ethyl acetate in hexanes gradient) to yield ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)- 6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetate (205 mg, 266 μmol, 93% yield) as yellow oil.
Synthesis of 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyπamino)-6-bromo-3-(Quinolin- 3-vOpyrazoloH .5-aipyrimidin-5-yl)cvclohexyl)acetonitrile:
Figure imgf000334_0002
2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(quinoiin-3- yl)pyrazoloti .5-alpyrimidin-5-yl)cyclohexyl)acetonitriie was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsilyi)ethoxy)methyl)amino)-6- bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-ajpyhmidin-5-yl)cyclohexyl)acetate, but with 2-(4- (7-{bis((2-(trimethylsilyl)ethoxy)methyl)annino)-3-{quinolin-3-yl)pyrazo!ot1,5- a]pyrimidin-5-yl)cyclohexyl)acetonitrile substituted ethyl 2-{4-{7-(bis((2- (tπmethy!silyl)ethoxy)methyl)amino)-3-(quinolin-3-yl)pyrazoIoE1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate.
Synthesis of 2-(4-(7-amino-6-bromo-3-(αuinolin-3-yl)pyrazolo[1 ,5-a1pyrimidin-5- yl)cvclohexyl)acetic acid:
Figure imgf000335_0001
To a 20 nrsL scintillation vial was charged ethyl 2-(4-{7-(bis((2-(trimethy!silyl)ethoxy)- methyl)amino)-6-bromo-3-(quinolin-3-yl)pyrazoio[1 ,5-a]pyrirrιidin-5- yl)cyclohexyl)acetate (100 mg, 130 μmol) and 2:1 THF:H2O (10 ml_). To this solution was added 1 N LiOH(aq) (200 μi_). The resulting reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo and the resulting residue was dissolved in TFA and stirred at room temperature for 1 hour. This solution was concentrated in vacuo and the residue was dissolved in 3:1 DMSO:MeCN. The crude product was purified via reverse-phase preparatory HPLC to yield 2-(4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyciohexyl)acetic acid (m+H = 480.1 , retention time = 3.95 min (isomer 1) and 4.03 min (isomer 2))
Synthesis of fraπs-2-(4-(7-amino-6-bromo-3-(αuinolin-3-v[)pyrazo]oi"1 ,5-alpyrimidin-5- yl)cvclohexyPacetic acid and c/s-2-(4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5- alpyrimidin-5-yl)cvclohexyl)acetic acid:
Figure imgf000336_0001
2-(4-(7-amino-6-bromo-3-(quinolin-3-yi)pyrazoio[1 ,5-a3pyrimidin-5-yl)cyclohexyl)acetic acid is taken up in 9:1 MeOH:TFA. The mixture is etuted through chiral column to yield two single isomers. (m+H = 480.1 , 10 min. RP HPLC Ret(isoi): 4.03 min., 10 min. RP HPLC Ret(iso2): 3.95 min.)
Synthesis of 2-(4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-alpyrimidin-5- vQcyclohexyl)acetonitrile:
Figure imgf000336_0002
2-(4-(7-amino-6-bromo-3-{quinolin-3-yi)pyrazolo[1 ,5-a3pyrimidin-5- yl)cyciohexyl)acetonitrile (44 μmol, 32 mg) is disolved in 1 mL TFA in a 20 mL scintillation viai. The reaction mixture is allowed to stir 1 hour at room temperature. The reaction mixture is concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield 2-(4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyciohexyi)acetonitrile as a two-isomer mixture. (m+H = 461.2, retention time = 4.13 min (isomer 1 ) and 4.18 min (isomer 2))
5-Trøns^-(fi H-tetrazol-5-yl)methyπcvclohexyl)-β-bromo-3-(quinolin-3-yl)pyrazoloπ .5- alpyrimidin-7-amine and 5-c/s-4-((1 H-tetrazoi-5-yl)methy0cvclohexyl)-6-bromo-3- (quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine:
Figure imgf000337_0001
To a solution of 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetonitrile (28 mg, 0.044 mmol) in dioxane {0.50 mL) was added azotributyltin azide (22 mg, 0.066 mmoi). The mixture was heated at 105 °C for 20 h. The tin reagent was removed by a short column and the fraction containing the desired product was collected and concentrated in vacuo. The residue was dissolved in CH3CN and NBS (2.0 mg) was added. The mixture was stirred at rt for 1 h and concentrated in vacuo. The residue was purified by prep-LC to give the fraπ-isomer and the c/s-isomer of the title compounds. LC/MS RT = 3.53 min (trans), 3.73 min (cis). Mass calculated for, M+H 504.12, observed 504.12.
By essentially the same procedures given in the complete synthesis of 2-(4-(7-amino- 6-bromo-S^quinolin-3-yl)pyrazoloII .S-alpyrimidin-5-yl)cyclohexyl)acetonitrile, but with cis/trans isomers separated on silica gel after hydrogen reduction. By essentially the same procedure given in Scheme 6, the compounds listed in Table 5 can be prepared.
Table 5
Figure imgf000337_0002
Figure imgf000338_0001
Figure imgf000339_0001
Figure imgf000340_0002
fΦfy-Amino-6-pyridin-Φyl-S-quinolin-S-yl-pyrazolori .5-alpyrimidin-5-yl)-cvclohexyll- acetic acid:
Figure imgf000340_0001
SCHEME-21 :
4-Pyridine boronic acid (0.156 mmoi, 20 mg), K2CO3 (0.234 mmol, 33 mg), and Pd(PPh3)4 (0.008 mmol, 10 mg) was added to a solution of (4-{7-[bis-(2- trimethylsilanyl-ethoxymethyl)-aminol-e-bromo-S-quinolin-S-yl-pyrazoloCI .5- a]pyrimidin-5-yl}-cyciohexyl)-acetic acid ethyl ester (0.078 mmol, 60 mg) in dioxane (1 mL). To this suspension was added distilled H2O (0.2 ml_). The resulting reaction mixture was stirred at 100° C under argon for 18 hours. The reaction mixture was concentrated in vacuo. The crude mixture was dissolved in 2:1 MeOH:H2O (1.5 mL) and was treated with 2N NaOH(aq) {0.5 mL). The resulting reaction mixture was stirred at room temperature for 18 hours. Aqueous hydrochloride solution (1.0 N, 2 ml) was added to the reaction mixture and the resulting solution was stirred at 65 °C for 2 h. The solution was concentrated in vacuo and purified by prep-LC to afford the title compound (3.6 mg) as a mixture of cis and trans isomers: LC/MS RT = 2.84 min. Mass calculated for, M+H 479.21 , observed 479.21. r4-(7-Amino-6-cvano-3-quinolin-3-yl-pyrazoion ,5-a1pyrimidin-5-yl)-cvclohexyl]-acetic acid
A degassed mixture of (4-{7-[bis-(2-trimethylsilanyl-ethoxymethyl)-amino3-6-bromo-3- quinolin-3-yl-pyrazolo[1 ,5-a]pyrimidin-5-yt}-cyclohexyl)-acetic acid ethyl ester {0.078 mmol, 60 mg), Bu3SnCN (50 mg, 0.156 mmoL), Pd[P{t-Bu)3]2 (8.3 mg, 0.016 mmoL) in Dioxane (1 mL) was heated at 100°C for 18 h. The reaction mixture was concentrated in vacuo. The crude mixture was dissolved in 2:1 MeOH:H2O (1.5 mL) and was treated with 2N NaOH(aq) (0.5 mL). The resulting reaction mixture was stirred at room temperature for 18 hours. Aqueous hydrochloride solution {1.0 N, 2 ml) was added to the reaction mixture and the resulting solution was stirred at 65 °C for 2 h. The solution was concentrated and purified by prep-LC to afford the title compound (8.8 mg) as a mixture of cis and trans isomers: LC/MS RT = 3.88 min. Mass calculated for, M+H 427.18, observed 427.18.
By essentially the same procedure given in Scheme 7, the compounds listed in Table 6 can be prepared.
Table 6
Figure imgf000341_0001
Figure imgf000342_0001
2-(4-(7-amino-6-bromo-3-(αuinolin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)cvclohexyl)-N- (methylsulfonvOacetamide
Figure imgf000343_0001
SCHEME-22
A mixture of 2-(4-(7-{bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(quinolin-3- yl)pyrazoio[1 ,5-a]pyrimidin-5-yi)cyclohexyl)acetic acid (60 mg, 0.081 mmo!), methanesuifonamide (12 mg, 0.122 mmol), 4-dimethy!aminopyridine (20 mg, 0.163 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (32 mg, 0.163 mmol) in dichloromethane (2.0 ml) was stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo. The crude mixture was dissolved in methanol (2 ml) and treated with 1 N hydrochloride solution (2 ml) at 65 °C for 2 h. The reaction solution was concentrated and purified by prep-LC to afford the title compound (two isomers, cis and trans). Isomer 1 : LC/MS RT = 3.63 min. Mass calculated for, M+H 557.09, observed 557.09. Isomer 2: LC/MS RT = 3.88 min. Mass calculated for, M+H 557.09, observed 557.09.
By essentially the same procedure given in Scheme 22, the compounds listed in Table 7 can be prepared.
TABLE- 7
Figure imgf000343_0002
Figure imgf000344_0002
Ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)m8thyl)amino)-3-(naphthalen-2- vnpyrazoion ,5-aipyrimidin-5-yl)cvclohexyl)acetate. LC/MS RT = 3.40 min {5 min method). Mass calculated for, M+H 689.38, observed 689.38.
Figure imgf000344_0001
2-(4-(7-Amino-6-bromo-3-(6-bromonaphthalen-2-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cvclohexyl)acetic acid . LC/MS RT = 6.01 min (10 min method). Mass calculated for, M+H 557.01 , observed 557.01.
Figure imgf000345_0001
2-(4-(7-amino-6-bromo-3-(6-bromonaphthalen-2-yl)pyrazolo[1 ,5-a]pyrimidin -5- yl)cyclohexyl)-N-(methylsulfonyl)acetamide. LC/MS RT = 5.86 min (10 min method). Mass calculated for, M+H 634.01 , observed 634.01 .
Figure imgf000345_0002
fS)-2-(f1s,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-alpyhmidin-5- yl)cvclohexyl)-2-fluoroacetic acid
Part-A
Ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)m8t{iyl)amino)pyrazolo[1 ,5-a1pyrimidin-5- yl)cvclohexylidene)-2-fluoroacetate
Figure imgf000345_0003
To a 250 mL roundbottom flask containing 80 ml. THF was charged sodium hydride (60% w/w in mineral oil, 12.86 mmol, 515 mg). This suspension was broken up in the sonicaid and then cooled to 0° C in icebath. Triethyl 2-fluoro-2-phosphonoacetate (12.86 mmol, 3.12 g) was taken up in THF (10 ml_) and added dropwise to stirring NaH suspension. The resulting solution was allowed to stir at 0° C for 10 minutes. 4- (7-(bis{(2-(trimethylsilyl)ethoxy)methyI)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexanone (6.43 mmol, 3.16 g) was taken up in THF (10 mL) and added dropwise to the stirring solution. The reaction was allowed to warm to room temperature and stir 18 hours. At 18 hours, the reaction was diluted with DCM (400 mL) and washed with H2O. The organic layer was collected and dried over NaaSO4. This solution was reduced in vacuo and purified via flash chromatography to yield the title compound (3.43 g, 5.92 mmol).
PART-B
Ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyπamino)pyrazoloπ ,5-alpyhmidin-5- yl)cvclohexylidene)-2-fluoroacetate
Figure imgf000346_0001
To a 50 mL roundbottom flask was charged ethanol (20 mL), ethyl 2-(4-(7-(bis((2- (trimethylsilyl)ethoxyJmethyl)aminoJpyrazoloCI ^-aJpyrimidin-5-yl)cyclohexylidene)-2- fiuoroacetate (1.99 mmol, 1.15 g), and 10% palladium on carbon (200 mg). The flask was sealed and degassed under vacuum. Hydrogen gas was then added via balloon and the reaction was allowed to stir at room temperature for 18 hours. At 18 hours the reaction was filtered through celite and the solvent reduced in vacuo. The title compound was then purified via flash chromatography on high-performance silica (829 mg). PART-C
(S)-ethyi 2-((1s.4R)-4-(7-(bis(f2-(trimethylsilyl)ethoxy)methynaminoV3- iodopyrazolofi ^-aipyrimidin-5-yl)cvclohexyl)-2-fluoroacetate
Figure imgf000347_0001
To a 50 ml_ roundbottom flask was charged acetonitrϋe (20 mL), ethyl 2-(4-(7-(bis({2- (trimethylsϋyl)ethoxyJmethyl)aminoJpyrazoioti .5-alpyrimidin-5-yllcyclohexylidene)-2- fluoroacetate (1.43 mmol, 828 mg), and N-lodosuccinimtde (1.57 mmol, 353 mg). The resulting solution was allowed to stir at room temperature for 18 hours. At 18 hours, the reaction was reduced in vacuo and purified via flash chromatography to yield the title compound (995 mg, 1.41 mmol). PART-D
(S)-ethyl 2-(f1s,4R)-4-(7-(bis(f2-(trimethylsilyl^ethoxy)methynaminoV3-(6- phenylpyridin-G-vOpyrazoloπ.5-aipyrimidin-5-vhcvclohexyπ-2-fluoroacetate
Figure imgf000347_0002
A pressure vial was charged with (S)-ethyl 2-((1s,4R)-4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)-2- fiuoroacetate (4.26 mmol, 3.01 g), 6-phenylpyridine-3-boronic acid pinacol ester (5.54 mmol, 1.56 g), potassium phosphate (12.78 mmol, 2.71 g), PdCI2(dppf)-CH2CI2 (0.43 mmol, 348 mg), and a solution of 9:1 1 ,4-dioxane:H20 (45 mL). The flask was flushed with argon and sealed. The reaction was stirred at 80° C for 18 hours. At 18 hours, the reaction was diluted with DCM (200 mL) and washed with H2O. The organic layer was collected and dried over Na2SO4. The resulting residue was purified via flash chromatography to yield the title compound (3.79 mmol, 2.78 g) PART-E fS)-ethyl 2-(f1s.4RV4-(7-(bisff2-(trimethylsilyl)ethoxy)methyl)amino)-e-bromo-3-(6- phenylpyridin-3-yl)pyrazolori .5-aipyrimidin-5-yl)cvclohexyl)-Z-fluoroacetate
Figure imgf000348_0001
To a 100 mL roundbottom flask was charged (S)-ethyl 2-{(1s,4R)-4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3-yI)pyrazo[o[1 ,5--a]pyrimidin-5- yl)cyclohexyl)-2-fluoroacetate (3.51 mmol, 2.58 g), acetonitrile {40 mL) and N- bromosuccinimide (3.87 mmol, 688 mg). The resulting solution was stirred at room temperature for 18 hours. At 18 hours, the solvent was removed in vacuo and the residue was purified via flash chromatography to yield the title compound (3.21 mmol, 2.61 g) PART-F f SVethyl 2-ff1 s.4RV4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazoloπ .5- alpyrimidin-5-yl)cvciohexyl)-2-fluoroacetate
Figure imgf000348_0002
To a 50 mL roundbottom flask was charged (S)-ethyl 2-((1s,4R)-4-(7-(bis{(2- {trimethylsily!)ethoxy)methyf)amino)--6-bromo-3-{6-phenylpyridin-3-yl)pyrazoio[1 ,5- a]pyrimidin-5-yi)cyclohexyl)-2-fluoroacetate {0.62 mmol, 500 mg}, DCM {10 mL), and TFA (10 mL). The resulting solution was stirred at room temperature for 30 minutes. At 30 minutes, the solvent was removed in vacuo and the titie compound was purified by reverse-phase preparatory HPLC (m+H = 552.20, retention time = 3.46 min). PART-G
(S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6-phenylpyhdin-3-yl)pyrazoloπ .5-a1pyrimidin-5- yl)cvclohexyl)-2-f[uoroacetic acid
Figure imgf000349_0001
To a 20 mL scintillation vial was charged (S)-ethy! 2-((1 s,4R)-4-(7-amino-6-bromo-3- (6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyciohexyl)-2-fluoroacetate as a formic acid salt (0.25 mmol, 150 mg), a solution of 2:1 THF:H2O (9 mL), and 1 N NaOH(aq) (0.50 mmol, 0.50 mL). This solution was stirred at room temperature for 2 hours. At 2 hours, the solvent was removed in vacuo and the residue was taken up in 2 mL 1 :1 MeCN:H2θ. This solution was frozen in liquid nitrogen and then pumped dry on lyophilizer to yield (S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-y[)cyclohexyl)-2-fiuoroacetic acid as a sodium salt (m+H = 524.08, retention time = 4.16 min).
The following compounds ( Table-7A) were synthesized using essentially the same procedures used for the total synthesis of (S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6- phenylpyridin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)-2-fluoroacetic acid:
TABLE-7A
Figure imgf000350_0002
Ethyl 7-(bis((2-(trimethylsilyl)ethoxy)methynamino)-5-((1 R,4s)-4-((S)-2-ethoxy-1 - fluoro-2-oxoethyl)cyclohexyl)-3-(6-phenylpyridin-3-yl)pγrazolo[1 ,5-a1pyrimidine-6- carboxylate
Figure imgf000350_0001
To a pressure viai was charged (S)-ethyl 2-((1 s,4R)-4-(7-(bis((2- {trimethylsilyl)ethoxy)methyl)amirιo)-6-brorno-3-(6-phenylpyridin-3-y!)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)-2-fluoroacetate (0.32 mmol, 263 mg), tetrakis(tripheπylphosphine)palladium(0) (0.032 mmol, 37 mg), 1 ,4-dioxane (5 mL), and tributyl(1-ethoxyvinyl)tin (0.97 mmol, 329 μl_). The vessel was flushed with argon, sealed, and the reaction stirred at 100° C for 18 hours. At 18 hours, the solvent was removed in vacuo and the residue purified via flash chromatography to yield the title compound.
(SV2-((1s,4R)-4-(6-acetyl-7-amino-3-(6-phenylpyridSn-3-yl)pyrazolo[1 ,5-aipyrimidin-5- vhcvclohexyl)-2-fluoroacetic acid
Figure imgf000351_0001
To a 50 mL roundbottom flask is charged ethyl 7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)-5-((1 R,4s)-4-((S)-2-ethoxy-1 -fluoro-2~ oxoethyl)cyclohexyl)-3-tβ-phenyipyridin-3-yl)pyrazoloti ^-alpyrimidine-6-carboxylate (0.29 mmol, 236 mg), a solution of 2:1 THF:H2O (9 mL), and 1 N LiOH(aq) (0.59 mmol, 0.59 mL). The resulting solution was stirred at room temperature for 2 hours. The reaction was then acidified to pH ~ 4 with 1 N HCI(aq) and the solvent removed in vacuo. This residue was dissolved in 10 mL 1 ,4-dioxane. To this solution was added H2O (3 mL) followed by 4N HChdioxane (5 mL). This solution was stirred at room temperature for 18 hours. After 18 hours, the solvent was removed in vacuo and the residue purified via reverse-phase HPLC to yield (S)-2-((1s,4R)-4-(6-acetyl-7-amino-3- (6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cycIohexyl)-2-fluoroacetic acid (m+H = 488.62, retention time = 3.71 min). fS^-(fisΛRM-(e-acetyl-y-amino-3-(e-(i -(a-hydroxyethyD-I H-pyrazol-Φyl^pyridin-3- yl)pyrazolof1 ,5-alpyrimidin-5-yl)cvclohexyl)-2-fluoroacetic acid
HChdioxaπe
Figure imgf000352_0001
(S)-2-((1s,4R)-4-(6-acetyl-7-amino-3-(6-(1 -(2-hydroxyethyl)-1 H-pyrazol-4-yl)pyridin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)-2-fluoroacetic acid was synthesized in the manner previously described. (m+H = 521.22, retention time = 3.01 min).
(S)-2-((1 s,4R)-4-(7-amino-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cvclohexyl)-2-fluoroacetic acid
Figure imgf000353_0001
To a 50 ml. roundbottom flask was charged (S)-ethyl 2-({1s,4R)-4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-(6-pheny[pyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl^-fiuoroacetate (0.27 mmol, 200 mg), 1 ,4-dioxane (5 nriL), H2O (1 mL), and 4N HCI in 1 ,4-dioxane. The resulting solution was stirred at room temperature for 5 days. At day 5, the solvent was removed in vacuo and the residue taken on without further purification.
(S)-2-((1 s,4R)-4-(7-amino-6-chloro-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-alpyrimidin-5- yl)cvclohexyl)-2-fluoroacetic acid
Figure imgf000353_0002
To a 50 mL roundbottom flask was charged (S)-2-((1s,4R)-4-(7-amino-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)-2-f!uoroacetic acid hydrochloride (0.27 mmol), DMF (8 mL), and N-chlorosuccinimide (0.30 mmol, 40 mg). The resulting solution was stirred at room temperature for 18 hours. At 18 hours, the solvent was removed in vacuo. The resulting residue was purified via reverse-phase HPLC to yield (S)-2-((1 s,4R)-4-(7-amino-6-chloro-3-(6-phenylpyridin-3- yi)pyrazolo[1 ,5-a]pyrimidin-5-yi)cyclohexyI)-2-fluoroacetic acid (m+H = 480.02, retention time = 4.02 min).
fS)-ethyi 2-((1s,4RV4-(7-(bisff2-(trimethyisilyl)ethoxy)methyl)amino)-3-(6- 5 chloropyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)cvclohexyl)-2-fluoroacetate
Figure imgf000354_0001
(S)-ethy! 2-((1 s,4R)-4-(7-(bis({2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-
I O chloropyridin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyciohexyl)-2-fluoroacetate is synthesized in a manner similar to the sythesis of (S)-ethyl 2-((1s,4R)-4-(7-(bis((2- (trimethyIsilyl)ethoxy)methyl)amino)-3-(6-phenyipyridin-3-yl)pyrazo!o[1 ,5-a]pyrimidin-5- yl)cyclohexyl)-2-fluoroacetate, but with 6-chloropyridine-3-boronic acid pinacol ester substituted for 6-phenylpyridine-3-boronic acid pinacol ester.
15 fS)-ethyl 2-(f1 s.4R)-4-(7-(bisff2-(trimethylsilyl)ethoxy)methynamino)-3-(6-(pyrimidin-2- yl)pyridin-3-yl)pyrazolof1 ,5-aiPyrimidin-5-yl)cvdohexyl)-2-fluoroacetate
Figure imgf000354_0002
20
To a 2-5 mL microwave vessel was charged (S)-ethyl 2-((1s,4R)-4-(7-(bis{(2- (trimethylsi[yl)ethoxy)methyl)amino)-3-(6-chloropyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cycIohexyl)-2-fluoroacetate (0.22 mmol, 150 mg), acetonitrile (3 mL), 2- (tributylstannyi)pyrimidine (0.65 mmoi, 240 mg), and tetrakis(triphenylphosphine)palladium{0) (0.02 mmol, 25 mg). The vessel was flushed with argon, sealed and heated to 150° C for 40 minutes in microwave synthesizer. Upon completion, the solvent was removed in vacuo and the residue purified by flash chromatography to yield the title compound.
(S)-2-(f1s.4R)-4-('7-amino-6-bromo-3-(6-(pyrimidin-2-yl)pyridin-3-yl)pyrazoloπ .5- a1pyrimidin-5-yl)cvclohexyl)-2-fluoroacetic acid
Figure imgf000355_0001
(S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6-(pyrimidin-2-y[)pyridin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)-2-fluoroacetic acid is synthesized in a manner similar to the sythesis of (S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3- yl)pyrazolo[1.5-aJpyrimidin-5-yl)cyclohexyl)-2-fluoroacetic acid (m+H = 524.80, retention time = 3.70 min).
The following compounds ( Table-7B) were synthesized using essentially the same procedures used for the total synthesis of (S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6- (pyrimidin-2-yl)pyridin-3-yi)pyrazolo[1,5-a]pyrimidin-5-yl)cyciohexyl)-2-fluoroacetic acid: TABLE-7B
Figure imgf000356_0001
Figure imgf000357_0001
Figure imgf000358_0001
Figure imgf000359_0001
2-(4-(4A5,5-tetramethyl-1.3.2-dioxaborolan-2-yl)-1 H-pyrazoi-1-yltethyl acetate
Figure imgf000360_0001
To a 20 ml_ pressure vial was charged pyrazole-4-boronic acid pinacoi ester (5.15 mmo!, 1.00 g) and DMF (10 ml_). NaH (60% w/w in mineral oil, 5.67 mmol, 227 mg) was then added portion-wise. The resulting solution was stirred at room temperature for 10 minutes. 2-bromoethyl acetate (5.67 mmol, 623 μl_) was then added dropwise. The vial was flushed with argon, sealed, and the reaction was stirred at 60° C for 18 hours. At 18 hours, the DMF was removed in vacuo, and the residue taken up in DCM. This suspension was filtered through celite and then purified via flash chromatography to yield the title compound.
2-(Φ-f4,4,5.5-tetramethyi-1.3.2-dioxaborolan-2-vh-1 H-pyrazoi-1-yl)ethanol
Figure imgf000361_0001
To a 50 mL roundbottom flask was charged 2-(4-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yI)-1 H-pyrazol-1-y!)ethyl acetate (2.11 mmol, 591 mg), a solution of 2:1 THFiH2O (15 mL), and 1 N LiOH(aq) (4.82 mmol, 4.82 mL). The resulting solution was stirred at room temperature for 48 hours. At 48 hours, the reaction was quenched with 1 N HCI(aq) (4.82 mmol, 4.82 mL) and the solvent removed in vacuo. The product was taken forward without further purification.
1 -(2-trideuteromethoxyethyl)-4-(4.4.5.5-tetramethyl-1 ,3,2-dioxaborolan-2-vh-1 H- pyrazoie
Figure imgf000361_0002
To a 20 mL pressure via! was 2-(4-(4,4,5,5-tetramethy!-1 ,3,2-dioxaborolan-2-y!)-1 H- pyrazol-1-yl)ethanol (2.1 1 mmol) and DMF (10 mL). NaH (60% w/w in mineral oil, 4.22 mmol, 169 mg) was added portion-wise and the resulting soiution was allowed to stir at room temperature for 10 minutes. At 10 minutes, iodomethane-d3 (4.22 mmoi, 263 μL) was added dropwise. The vial was sealed and the reaction was stirred at 80° C for 48 hours. At 48 hours, the solvent was removed in vacuo. The residue was taken up in DCM and filtered through celite. The title compound was then purified via fiash chromatography.
2-methy)-1 -(4-(4.4.5.5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-1 H-ρyrazoi-1 -vnpropan-2- ol
Figure imgf000362_0001
To a 20 mL pressure vial was charged pyrazole-4-boronic acid pinacol ester (5.15 mmol, 1.00 g), DMF (10 mL), and Cs2CO3 (5.67 mmoi, 1.85 g). To this suspension was added isobutylene oxide (15.45 mmol, 1.38 mL). The vial was flushed with argon and sealed. The reaction was stirred 18 hours at 80° C. After 18 hours, the reaction was filtered through celite and the solvent removed in vacuo. This residue was pumped dry under high vacuum and then purified via flash chromatography to yield the title compound.
1 -trideuteromethyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-ylV1 H-pyrazole
Figure imgf000362_0002
1 -trideuteromethyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-1 H-pyrazo!e was synthesized in the manner previously described. 3-(6-phenylpyridin-3-ylV5-(1 H-Pyrazol-4-yl)-N.N-bis((2- (trimethyisilyl)ethoxy)methyl)pyrazolo[1,5-a1pyrimidin-7-amine
Figure imgf000363_0001
3-(6-phenylpyridin-3-yl)-5-(1 H-pyrazot-4-yl)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine was synthesized in a manner similar to the sythesis of (S)-ethyl 2-((1s,4R)-4-{7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenyipyridin-3-yl)pyrazo!o[1 ,5-a]pyrimidin-5- yl)cyciohexyl)-2-fluoroacetate.
2-(4-(7-(bis{(2-(trimethylsilyl)ethoxy)methyl)anninoV3-(6-phenylpyridin-3- vOpyrazolofi ,5-aipyrimidin-5-vP-1 H-pyrazol-1 -yl)ethyl acetate
Figure imgf000363_0002
To a 2-5 mL pressure vial was charged 3-(6-phenylpyridin-3-yt)-5-(1H-pyrazol-4-yl)- N,N-bis((2-(trimethylsily!)ethoxy)methyl)pyrazolo[1 f5-a]pyrimidin-7-amine (0.38 mmol, 230 mg), DMF (4 mL), and sodium hydride (40% w/w in mineral oil, 0.56 mmol, 23 mg). This suspension was sonicaided and allowed to stir at room temperature for 15 minutes. At 15 minutes, 2-bromoethyl acetate (0.56 mmol, 62 μl_). The vial was then sealed and the reaction was stirred at 80° C for 18 hours. After 18 hours, the solvent was removed in vacuo, and the resulting residue was purified by flash chromatography to yield the title compound.
1 -(7-amino-5-(1 -(2-hydroxyethyl)-1 H-pyrazol-4-yl)~3-(6-phenylpyridin-3- vQpyrazoio[1 ,5-alpyrimidin-6-yl)ethanone
Figure imgf000364_0001
Figure imgf000364_0002
1 -(7-amino-5-(1 -(2-hydroxyethyl)-1 H-pyrazol-4-y!)-3-(6-phenylpyridin-3- yl)pyrazolo[1 ,5-a]pyrimidin-6-yl)ethanone was synthesized in a manner similar to the sythesis of {S)-2-((1s,4R)-4-(6-acetyl-7-amino-3-(6-phenylpyπdin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)-2-fluoroacetic acid (m+H = 440.64, retention time = 2.95 min).
Ethyl 2-(4-(7-(bisf(2-(trimethylsilyl)ethoxy)methyi')aminoV3-(1 -phenyl- 1H-pyrazoi-4- vQpyrazolon .5-alpyrimidin-5-vOcvclohexynacetate
Figure imgf000365_0001
Ethyl 2-(4-(7-{bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(1 -phenyl-1 H-pyrazol-4- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetate was synthesized in a manner similar to the sythesis of (S)-ethyl 2-((1s,4R)-4-(7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)-3-{6-phenylpyridin-3-yi)pyrazoto[1 ,5-a]pyrimidin-5- yl)cyclohexyl)-2-fluoroacetate.
2-(4-(7-amino-6-bromo-3-(1 -phenyl-1 H-pyrazol-4-yl)pyrazoloH ,5-a1pyrimidin-5- yl)cvclohexyl)acetic acid
Figure imgf000365_0002
2-(4-(7-amino-6-bromo-3-(1 -phenyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5-a]pyrimidin-5- yi)cyciohexyl)acetic acid was synthesized in the manner previously described. (m+H = 495.57, retention time = 4.78 min (isomer-!) and 4.84 min (isomer 2)). 0350
365
(SV2-((1 s,4RV4-(7-amtno-6-bromo-3-(1 -phenyl-1 H-pyrazol-4-vhpyrazoloH ,5- aipyrimidin-5-yl)cvclohexyl)-2-fiuoroacetic acid
Figure imgf000366_0001
(S)-2-((1s,4R)-4-(7-aminθ"6-bromo-3-(1-phenyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5- a]pyrimidin-5-yi)cyclohexyl)-2-fluoroacetic acid was synthesized in the manner similar to the synthesis of 2-{4-(7-amino-6-bromo-3-(1-phenyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)acetic acid. (m+H = 513.54, retention time = 4.81 min).
2-(4-(6-acetyl-7-amino-3-(1 -phenyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5-alpyrimidin-5- yl)cvciohexyl)acetic acid
Figure imgf000366_0002
2-(4-{6-acetyl-7-amino-3-(1 -phenyl-1 H-pyrazol-4-yl)pyrazo!o[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetic acid was synthesized in the manner previously described. {m+H = 459.66, retention time = 4.58 min).
(SV2-((1 s,4R)-4-(6-acetyl-7-amsno-3-(1 -phenyl-1 H-pyrazoi-4-vQpyrazolo[1.5- alpyrimidin-5-vOcvclohexyπ-2-fluoroacetic acid
Figure imgf000367_0001
(S)-2-((1 s,4R)-4~(6-acetyl-7-amtno-3-(1 -phenyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)-2-fluoroacetic acid was synthesized in the manner similar to the synthesis of 2-(4-(6-acetyl-7-amino-3-(1 -phenyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)acetic acid. {m+H = 477.64, retention time = 4.53 min).
Ethyl 6-(7-(bisf(2-(trimethylsilv[)ethoxy^methyl)amino)pyrazolo[1 ,5-alpyrimidin-5- yl)spirof2.5'loctane-1-carboxylate
Figure imgf000367_0002
To 50 mL roundbottom flask containing trimethylsuifoxonium iodide (3.57 mmol, 785 mg) in DMSO (8 mL) was added potassium tert-butoxide (3.57 mmol, 401 mg). This suspension was allowed to stir at room temperature for 3 hours. After 3 hours, ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexylidene)acetate (1.78 mmol, 1.00 g) was added in 2 mL DMSO. The resulting solution was stirred at room temperature for 18 hours. After 18 hours, the reaction was diluted with saturated NaC!(aq) (30 mL). The pH was adjusted to ~7 with saturated NH4CI(aq) and extracted with DCM (50 mL) 4 times. The combined organics are dried over Na2S0_j and reduced in vacuo. The resulting residue was purified via flash chromatography to yield the title compound. Ethyl 6-( 7-(bis(f2-(trimethyisilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-aiPyrimidin-5-y1H ■ f!uorospiro[2.51octane-1 -carboxylate
Figure imgf000368_0001
Ethyl 6-(7-(bis((2-(trinrtethylsily!)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5-y!)-1 ■ fluorospiro[2.5]octane-1 -carboxylate was synthesized in the manner similar to the synthesis of ethyl 6-(7-(bis((2-(trimethytsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)spiro[2.5]octane-1 -carboxylate.
6-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-alpynmidin-5- yl)spiro[2.5]octane-1 -carboxylic acid
Figure imgf000368_0002
6-(7-amino-6-bromo-3-<6-phenylpyridin-3-yl)pyrazolo[1 ,5~a]pyrimidin-5- yl)spiro[2.5]octane-1 -carboxylic acid was synthesized in the manner previously described. {m+H = 417.86, retention time = 4.41 min).
6-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazoio[1 ,5-alpyrimidin-5-yl)-1- f Iuorospirof2.51octane-1 -carboxylic acid
Figure imgf000369_0001
6-(7-amino-6-bromo-3-(6-pheny!pyridin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5-yI)-1 - fluorospiro[2.5]octane-1 -carboxylic acid was synthesized in the manner previously described. (m+H = 435.86, retention time = 4.40 min).
ethyl 2-acetoxy-2-(4-(7-(bis((2-(trimethylsilyltethoxy)methy0amino)pyrazoloH ,5- aipyrimidin-5-yl)cvclohexylidene)acetate
Figure imgf000369_0002
To a 100 mL roundbottom flask was charged ethyl 2-acetoxy-2- (diethoxyphosphoryl)acetate (11.21 mmol, 3.16 g), LiCI (1 1.21 mmoi, 475 mg), and THF (25 mL). The flask was then flushed with argon, sealed, and cooled to -78° C in a dry ice/lPA bath. Added dropwise to this solution was a solution of 1 ,1 ,3,3- tetramethylguanidine (11.21 mmol. 1.41 g) in THF (5 mL). The resulting solution was allowed to stir at -78° C for 30 minutes. At 30 minutes, 4-{7-(bis((2- (trimethylS!lyl)ethoxy)methyl)am!no)pyrazolo[1 ,5-a]pynmidin-5-yl)cyclohexanone (5.09 mmol, 2.50 g) was added in THF (3 mL). The reaction was then allowed to siowly warm to room temperature and then stirred for 48 hours. After 48 hours, the reaction was diluted with DCM (100 mL) and washed with H2O (50 mL). The organic layer was collected, dried over Na2SO4, and reduced in vacuo. The resulting residue was purified via flash chromatography to yield the title compound.
Ethyl 2-acetoxy-2-(4-(7-(bisf(2-(trimethylsiiyl)ethoxy)methynamino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)cvdohexylidene)acetate
Figure imgf000370_0001
Ethyl 2-acetoxy-2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyc!ohexylidene)acetate was synthesized in the manner previously described.
2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenylpyridin-3- vOpyrazoioπ .5-a1pyrimidin-5-yl)cvclohexyl)-2-oxoacetic acid
Figure imgf000371_0001
To a 50 mL roundbottom flask was charged ethyl 2-acetoxy-2-(4-(7-(bis((2-
{trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5- alpyrimidin-5-yl)cyclohexylideneJacetate (0.47 mmol, 400 mg), a solution of 2:1 THF:H2O (15 mL), and 1 N LiOH(aq) (1.18 mmol, 1.18 mL). The resulting solution was stirred at room temperature for 18 hours. After 18 hours, the reaction was quenched with 1 N HCI(aq) (1.18 mmol, 1.18 mL) and the solvent removed in vacuo. The resulting residue was taken on without further purification.
(RV2-(f1 r.4R)-4-(7-(bisα2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6- phenylpyπdin-3-yl)pyrazolori ,5-alpyrimidin-5-yl)cvclohexylV2-hydroxyacetic acid
Figure imgf000372_0001
To a 50 mL roundbottom fiask was charged 2-{4-(7-(bis((2- (tπmethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6--phenylpyridin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yi)cyclohexyl)-2-oxoacetic acid (0.47 mmol) and MeOH (10 mL). To this solution was charged NaBH4 (0.71 mmol, 27 mg). This solution was allowed to stir at room temperature for 18 hours. After 18 hours, the reaction was quenched with saturated NH4CI(aq) {10 mL) and extracted with DCM twice. The combined organics were dried over Na≥SCλi, and the solvent removed in vacuo. The compound was taken on without further purification.
(R)-1-(f1 r.4R)-4-(7-(bisff2-(trimethylsiivnethoxy)methyl)amino)-3-(6-phenylpyridin-3- yl)pyrazolori ,5-aipyrimidin-5-yl)cyclohexyl)ethane-1 ,2-diol
Figure imgf000372_0002
To a 50 mL roundbottom flask was charged (R)-2-((1 r,4R)-4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenyIpyridin-3"yl)pyrazolo[1 ,5- a]pyrimidin-5-yi)cyclohexyl)-2-hydroxyacetic acid (0.47 mmol) and THF (20 mL). This solution was cooled to 0° C in an ice bath. Lithium aluminum hydride (1.0 M in hexanes, 2.06 mmol, 2.06 mL) was added dropwise. The reaction is then allowed to slowly warm to room temperature and then stirred for 30 hours. After 30 hours, the reaction was quenched with EtOAc (20 mL) and washed with H2O. The organic layer was dried over Na2SO4, and the solvent removed in vacuo. The product was taken on without further purification.
(FO-1 -((1 r,4R)-4-{7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5- yl)cyclohexyl)ethane-1 ,2-diol
Figure imgf000373_0001
(R)-1-((1 r,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)ethane-1 ,2-diol was synthesized in the manner previously described. (m+H = 507.84, retention time = 3.56 min).
1 -(7-amino-5-((1 R,4r)-4-(f RV 1 ^-dihydroxyethyncvclohexyl)-3-(e-phenylpyridin-3- vθpyrazolori ,5-a1pyrimidin-6-yl)ethanone
Figure imgf000373_0002
1 -(7-amino-5-((1 R,4r)-4-((R)-1 ^-dihydroxyethyl)cyclohexyl)-3-^-phenyipyridin-3- yl)pyrazoio[1 ,5-a]pyrimidin-6-yl)ethanone was synthesized in the manner previously described. (m+H = 471.98, retention time = 3.45 min). (R)-2-(f1 rτ4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5- yl)cyclohexyl)-2-hydroxyacetic acid
Figure imgf000374_0001
(R)-2-((1 r,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexy!)-2-hydroxyacetic acid was synthesized in the manner previously described. (m+H = 522.06, retention time = 3.51 min).
2-((1 rT4r)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo|-1.5-a1pyrimidin-5- yl)cvclohexyl)-2-oxoacetic acid
Figure imgf000374_0002
2-((1 r,4r)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)-2-oxoacetic acid was synthesized in the manner previously described. (m+H = 520.05, retention time = 3.79 min).
(SVethyl 2-acetoxy-2-((1 s,4R)-4-(7-(bisff2-(trimethylsilvnethoxy)methyπamino)-3-(6- phenylpyridin-3-yl)pyrazolofi .5-aipyrimidin-5-yl)cvclohexyQacetate
Figure imgf000375_0001
To a 50 ml_ roundbottom flask was charged ethyl 2-acetoxy-2-(4-(7-(bis((2- (trimethylstlyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyiidene)acetate (0.47 mmol, 400 mg), ethanol (15 ml_), di- isopropylethyl amine (0.50 mmol, 88 μL), and Raney 2800 nickel (slurry in H2O, 100 mg). The flask was sealed and degassed under vacuum for 15 minutes. After 15 minutes, the hydrogen gas was added in balloon and the reaction stirred at room temperature for 18 hours. After 18 hours, the reaction was filtered through celite,
10 being careful to not dry out Raney nickel. The filtrate was reduced in vacuo and the resulting residue was purified via flash chromatography to yield the title compound.
(S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolof1 ,5-alpyrimidin-5- vOcycloheχγQ-2-hydroxyacetic acid
\5
Figure imgf000375_0002
(S)-2-((1s,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)~2-hydroxyacetic acid was synthesized in the manner previously 0 described. (m+H = 522.10, retention time = 2.68 min).
(R)-methyl 2-(f1 r,4R)-4-(7-(b!S((2-(trimethylsilyl)ethoxy)methyπamino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolori .5-alPyrimidin-5-vncvciohexyl)-2-methoxyacetate
Figure imgf000376_0001
To a 50 ml_ roundbottom flask was charged (R)-2-((1 r,4R)-4-(7-(bis((2- (trimethylsi!yl)ethoxy)methyl)anrιino)-6-bromo-3-(6-phenylpyridin-3-yl)pyrazoio[1 ,5- a]pyrimidin-5-yI)cyclohexyl)-2-hydroxyacetic acid (0.40 mmol), MeOH (15 m!_), and TMS diazomethane (2.0 M in hexanes, 40 mmol, 20 mL). This solution was allowed to stir at room temperature for 72 hours. After 72 hours, the solvent was removed in vacuo and the residue was purified via flash chromatography to yield the title compound.
(R)-2-((1 rτ4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolori ,5-a1pyrimidin-5- yl)cvclohexyl)-2-methoxyacetic acid
Figure imgf000376_0002
(R)-2-((1 r,4R)-4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)-2-methoxyacetic acid was synthesized in the manner previously described. (m+H = 536.56, retention time = 3.84 min).
2-(4-(7-amino-3-(6-phenylpyridin-3-yl)pyrazolof1 ,5-aipyrimidin-5- vOcyclohexyl)acetohydrazide
Figure imgf000377_0001
To a 2-5 mL pressure vial was charged ethyl 2-(4-(7-(bis{(2- (trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)acetate (0.12 rnrnol, 95 mg), DMF (1 mL). and hydrazine monohydrate (1 mL). The pressure vial was sealed and heated to 80° C for 18 hours. After 18 hours, the solvents were removed in vacuo and the product taken on without further purification.
5-((4-(7-amino-3-(6-phenylpyridin-3-yl)pyrazoloπ .5-aipyrimidin-5- yl)cvciohexyl)methyiyi .3,4-oxadiazol-2(3H)-one
Figure imgf000377_0002
To a 20 mL scintillation vial was charged 2-{4-(7-amino-3-(6-phenylρyridin-3- yl)pyrazoloti .5-atøyrimidin-5-yl)cyclohexyl)acetohydrazide (0.12 mmol), THF (5 mL), and 1 ,1'-carbonyidiimidazo!e (0.12 mmol, 25 mg). The resulting solution was stirred at room temperature for 2 hours. After 2 hours, the solvent was removed in vacuo and the residue taken up in 3:1 DMSO: MeCN. The solids are removed via centrifugation, and the title compound was purified via reverse-phase HPLC.
5-((4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolof1.5-aipyrimidin-5- yl)cvclohexyl)methyl)-1.3.4-oxadiazol-2(3HVone
Figure imgf000378_0001
To a 20 mL scintillation vial was charged 5-((4-(7-amino-3-(6-phenylpyridin-3- yl)pyrazoio[1 ,5-a]pyhmidin-5-yt)cyclohexyl)methyl)-1 ,3,4-oxadiazol-2(3H)-one as a trifluoroacetate salt (0.035 mmol, 20 mg), MeCN (2 mL) and N-bromosuccinimide (0.035 mmol, 6.3 mg). The resulting solution was stirred at room temperature for 1 hour. After 1 hour, the solvent was removed in vacuo and the resulting residue was purified via reverse-phase HPLC to yield 5-((4-(7-amino-6-bromo-3-(6-phenylpyridin-3- yl)pyrazolo[1.5-alpyrimidin-5-yl)cyclohexyl)methylJ-i ,3,4-oxadiazol-2(3H)-one (m+H = 546.57, retention time = 4.07 min).
4-ftert-butoxycarboπylamino)cvclorιex-1-enyl trifluoromethanesulfonate
Figure imgf000378_0002
(Step 2)
To a 100 mL roundbottom flask was charged 4-N-Boc-aminocyclohexanone (4.69 mmol,
1.0Og) and THF (15 mL). The flask was flushed with argon and sealed with a rubber septum. The solution was then cooled to -78° C in a dry ice/isopropanol bath. LiHMDS (1.0 M in hexanes, 9.61 mmol, 9.61 mL) was then added dropwise. After addition, the resulting solution was stirred at -78° C for 45 minutes. At this point, 2-[NIN-Bis(trifiuoromethanesulfonyl)amino]- 5-chloropyridine (5.16 mmol, 2.03 g) was taken up in THF (5 mL) and added dropwise to stirring solution at -78° C. The resulting solution was allowed to gently warm to room temperature over 1 hour. At 1 hour, the reaction was quenched with H2O and extracted with DCM (x2). The combined organics were dried over Na2SO4 and the solvent them removed in vacuo. The residue was then purified via flash chromatography to yield the title compound.
4-(tert-butoxycarbonylamino)cvdohex-1-enyl trifluoromethanesulfonate
Figure imgf000379_0001
To a 10-20 mL pressure vial was charged 4-(tert-butoxycarbonylamino)cyclohex-1- enyi trifluoromethanesulfonate (1.15 mmol, 397 mg), 1 ,4-dioxane (8 mL), bis(pinacolato)diboron (1.72 mmol, 438 mg), potassium acetate (3.45 mmol, 339 mg), and PdCI2(dppf) ■ CH2Cl2 (1 15 μmol, 94 mg). The via! was flushed with argon, sealed, and heated to 80° C for 5 hours. After 5 hours, the solvent removed in vacuo and the residue was taken up in DCM. This suspension was filtered through celite and then purified via flash chromatography to yield the title compound.
tert-butyl 4-(7-(bisff2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolof1 ,5-alpyrimidin-5- vOcvclohex-3-enylcarbamate
Figure imgf000380_0001
tert-butyl 4-(7-(bis({2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohex-^-enylcarbamate was synthesized in the manner previously described.
tert-fautyl f 1 s,4s)-4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolori ,5- alpyrimidin-5-vOcyclohexylcarbamate and tert-butyl (1 r,4r)-4-(7-(bis((2- ftrimethylsilyl)ethoxy^methyπamino^pyrazolofi .5-aiPyrimidin-5-yl)cvclohexylcarbamate
Figure imgf000380_0002
To a 50 mL roundbottom flask was charged tert-butyl 4-{7-{bis{(2- (trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohex-3- enylcarbamate (0.67 mmoi, 396 irng), ethanol (15 mL), and 10% palladium on carbon (100 mg). The flask was flushed with argon, sealed, and degassed under vacuum. Hydrogen gas was added in a balloon and the reaction was allowed to stir at room temperature under a hydrogen atmosphere for 18 hours. After 18 hours, the reaction was filtered though ceiite. The solvent was removed in vacuo and the resulting residue was purified to yield the title compounds (isomer 1 = cis, isomer 2 = trans).
5-(dsΛsM-aminocyclohexyl)-e-bromo-3-(6-phenylpyridin-3-yl)pyrazoloπ .5- aipyrimidin-7-amine
Figure imgf000381_0001
5-tCis^sJ-Φaminocyclohexyl)-6-bromo-3-tβ-phenylpyridin-3-yl)pyrazoloti .5- a]pyrimidin-7-amine was synthesized in the manner previously described.
(R)-N-(f1s,4S)-4-(7-amino-6-brQmo-3-(6-phenylpyridin-3-yl)pyrazolof1 ,5-alpyrimidin-5- vOcvclohexyl)-2-hydroxypropanamide
Figure imgf000381_0002
To a 20 mL scintillation vial was charged 5-((1 s,4s)-4-aminocyclohexyl)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazo!o[1 ,5-a]pyrimidin-7-amine as a hydrochloride salt (37 μmol), DMF (2 mL), D-!actic acid (0.08 mmol, 7.2 mg), EDC (0.08 mmol, 15.4 mg), HOBt (0.08 mmol, 10.8 mg), and DIEA (0.15 mmol, 26 μL). The resulting solution was stirred at room temperature for 4 hours. After 4 hours, the solvent was removed in vacuo and the residue purified via reverse-phase HPLC to yield (R)-N-((1 s,4S)-4-(7- amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazoIo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)-2- hydroxypropanamide (m+H = 534.74, retention time = 3.82 min).
The following compound ( Table-7C) was synthesized using essentially the same procedures used for the total synthesis of (R)-N-((1 s,4S)-4-(7-amino-6-bromo-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)-2-hydroxypropanamide:
TABLE-7C
Figure imgf000382_0001
(R)-N-((1s,4S)-4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)-2-hydroxypropanamide
Figure imgf000383_0001
{R)-N-((1s,4S)-4-(6-acetyl-7-amino-3-(6-pheny!pyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- ylJcyclohexyl)-2-hydroxypropanamide was synthesized in the manner previously described (m+H = 499.11 , retention time = 3.68 min).
The following compounds( Tabie-7D) were synthesized using essentially the same procedures used for the total synthesis of (R)-N-((1 s,4S)-4-(6-acetyl-7-amino-3-(6- phenylpyridin-3-yl)pyrazoloti .5-alpyrimidin-5-yl)cyclohexyl^-hydroxypropanamide:
TABLE-7D
Figure imgf000383_0002
Figure imgf000384_0002
Synthesis of 1 -(7-amino-5-((1 r,4r)-4-( methylsulfonylmethyl)cvclohexyπ-3-(1 -phenyl- 1 H-pyrazol-4-yl)pyrazoloM ,5-atoyrimidin-6-yl)ethanone:
Figure imgf000384_0001
SCHEME- 23 Part A: (lr.4r)-tert-butyl 4-(7-(bisf(2-(trimethylsilyl)ethoxy)methyl)amirLθ)-3-("l-phenyl-1H-pyrazol-4- yl)pyrazolori.5-alpyrimidin^-yl)cyclohexanecarboxyiate:
Compound, (lr,4r)-tert- butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(l-phenyl- 1H-pyrazol-4-y[)pyrazolo[1,5-a]pyrimidin-5-yl)cyclohexanecarboxylate was prepared from compound (lr,4r)-tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyI)amino)-3- iodopyrazolo[1,5-a]pyrimidin-5-yl)cyclohexanecarboxylate and 1 -phenyl-4-(4,4,5,5- tetramethyl- l,3,2-dioxaborolan-2-yl)-lH-pyrazoIe using the coupling conditions described in Part G of Example 8 (68%). ΗPLC-MS TR = 3.13 min (UV 254 nm, 5 min method); mass calculated for formula C38H58N6O4Si2718.4, observed LCMS m/z 719.3 (M+H).
Part B:
((1 r,4r)-4-(7-(bisf(2-(trimethylsilyl)ethoxy)methyl)amino)-3-(1 -phenyl-1 H-pyrazoi-4- yl)pyrazoiof1 ,5-alpyrimidin-5-yl)cvclohexyl)methanoi:
To a solution of compound (lr,4r)-tert-butyl 4-(7-(bis((2-
(trimethylsiIyl)ethoxy)methyl)amino)-3-(l-phenyl-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidin- 5-yl)cyclohexanecarboxylate (1.68 mmol, 1.21 g) in THF (20 mL) at 0 °C was added lithium aluminum hydride solution (1.68 mL, 2.0 M in THF). The resulting reaction mixture was slowly warmed to rt and stirred for 1 h. The reaction quenched with 3 N NaOH and filtered. The filtrate and washes were combined and concentrated. The crude product was purified by a SiO2 column (0-40% EtOAc/Hexanes, Rf = 0.7 in 50% EtOAc) to afford compound, ((lr,4r)-4- (7-(bis((2-(trimethylsily0ethoxy)methyl)amino)-3-(l-phenyl-1H-pyrazol-4-yl)pyrazolo[1,5- a]pyrimidin-5-yl)cyclohexyl)methanol, as a pale yellow solid (923 mg, 85%). HPLC-MS TR = 3.08 min (UV 254 nm, 5 min method); mass calculated for formula C34H52N6O3Si2648.4, observed LCMS m/z 649.3 (M+H).
Part C: f(1 r,4r)-4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(1-phenyl-1H-pyrazol-4- yl)pyrazolofi .5-alPyrimidin-5-yl)cvciohexyl)methyl methanesulfonate:
To a solution of compound, ((lr,4r)-4-(7-(bis((2-(trimethyIsilyl)ethoxy)methyl)amino)-3-(l- phenyI-1H-pyrazoi-4-yi)pyrazolo[1,5-a]pyrimidin-5-yl)cycIohexyl)methanol (1.42 mmol, 920 mg) in DCM (10 mL) was added TEA (4.26 mmol, 0.594 mL), followed by MsCl (2.13 mmol, 0.165 mL) at 0 °C and stirred for 1 h. The reaction mixture was washed with H2O and brine, dried over Na2SO4, and concentrated. The crude product was purified by a SiO2 column (0- 30% EtOAc/Hexanes, Rf = 0.8 in 50% EtOAc) to afford compound, ((lr,4r)-4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-(i-phenyl-1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidin- 5-yl)cyclohexyl)methyl methanesulfonate, as a pale yellow oil (949 rag, 92%).
Part D: 5-(fi rΛ^^-(methylthiomethyDcvclohexyπ-3-(i -phenyl-1H-pyrazol^-vh-N.N-bisffa- (trimethylsilyl)ethoxy)iinethyl)pyrazolo[1,5-a1pyrimidin-7-amine:
A mixture compound(( 1 r,4r) -4-(7-(b is((2-(trimethy lsilyi)ethoxy)methy!)amino)-3-(l -phenyl - 1H-pyrazol-4-yl)pyrazolo[l,5-a]pyrimidin-5-yi)cyciohexyl)methyl methanesulfonate (1.31 mmol, 949 mg) and NaSMe (2.61 mmol, 183 mg) in DMF (5 mL) was heated at 80 °C for 3 d. The reaction mixture was diluted with EtOAc, washed with H2O and brine, dried over Na2SO4, and concentrated. The crude product was purified by a SiO2 column (0-20% EtOAc/Hexanes, Rf = 0.5 in 20% EtOAc) to afford compound, 5-((lr,4r)-4-(methylthiomethyl)cyclohexyl)-3-(l- phenyl-1H-pyrazol-4-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin- 7-amine, as a pale yellow oil (804 mg, 90%). HPLC-MS TR = 3.59 min (UV 254 nm, 5 min method); mass calculated for formuia C35H54N6O2SSi2 678.4, observed LCMS m/z 679.3 (M+H).
Part E:
5-((1r,4r)-4-(mθthyisulfonylmethyl)cvclohexyl)-3-(1-phenyl-1 H-pyrazol-4-yl)-N,N-bisf(2- (trimethylsiiyl)ethoxy)methyl)pyrazolon ,5-a1pyrimiciin-7-amine:
To a solution of compound, 5-((l r,4r)-4-(methylthiomethyl)cyclohexyl)-3-(l-phenyl-1H- pyrazol-4-yI)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (1.18 mmol, 804 mg) in DCM (10 mL) was added mCPBA (2.83 mmol, 699 mg) and the resulting mixture was stirred at rt for overnight. The reaction was quenched with Na2S2O3 (aq.) and diluted with DCM (10 mL). The separated organic layer was washed with NaHCO3 (2*) and brine, dried over Na2SO4, and concentrated. The crude product was purified by a SiO2 column (0-40% EtO Ac/He xanes, Rf = 0.45 in 50% EtOAc) to afford compound, 5-((lr,4r)-4- (methylsulfonylmethyl)cycϊohexy])-3-(l -phenyl- 1H-pyrazoI-4-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoIo[1,5-a]pyrimidin-7-amine, as a pale yellow oil (446 mg, 53%). HPLC-MS TR = 2.99 min (UV 254 nm, 5 min method); mass calculated for formula C35H54N6O4SSi2710.3, observed LCMS m/z 71 1.2 (M+H).
Part F:
1 -(7-amino-5-((1 r,4r)-4-(methylsulfonylmethyl)cyciohexyl)-3-(1 -phenyl-1 H-pyrazol-4- yl)pyrazolori ,5-aipyrimidin-6-vQethanone:
By essentially the same procedures given in Preparative earliar Example compound, l-(7- amino-5-((lr,4r)-4-(methylsulfonylmethyl)cyclohexyl)-3-(l-phenyl-iH-pyrazol-4- yl)pyrazolo[1,5-a]pyrimidm-6-yI)ethanone was prepared from compound, 5-((lr,4r)-4- (methylsulfonylmethyl)cyclohexyl)-3-(l-phenyI-1H-pyrazol-4-yl)-N,N-bis((2- (trimethylsilyi)ethoxy)methy!)pyrazolo[1,5-a]pyrimidin-7-arnine. HPLC-MS TR = 4.64 min (UV 254 nm, 10 min method); mass calculated for formula C2SH28N6O3S, 492.2, observed LCMS m/z 493.1 (M+H).
Figure imgf000388_0001
SCHEME- 24
Synthesis of ethyl 1 ,4-dioxaspiro[4.51decane-7-carboxyiate
Figure imgf000388_0002
Ethyl 1 ,4-dioxaspiro[4.5]decane-7-carboxylate was synthesized in a manner similar to the synthesis of ethyl 1 ,4-dioxasptro[4.5]decane-8-carboxyiate, but with ethyl 3- oxocyclohexanecarboxylate substituted for ethyl 4-oxocyclohexanecarboxylate.
Synthesis of ethyl 1 ,4-dioxaspiro[4.41nonane-7-carboxylate
Figure imgf000389_0001
Ethyl 1 ,4-dioxaspiro[4.4]nonane-7-carboxylate was synthesized in a manner similar to the synthesis of ethyl 1 ,4-dioxaspiro[4.5]decane-8-carboxylate, but with ethyl 3- oxocyclopentanecarboxylate substituted for ethyl 4-oxocyclohexanecarboxytate.
Synthesis of 1 ,4-dioxaspirof4.51decane"7-carboxylic acid
Figure imgf000389_0002
1 ,4-Dioxaspiro[4.5]decane-7-carboxylic acid was synthesized in a manner similar to the synthesis of 1 ,4-dioxaspiro[4.5]decane-8-carboxyiic acid, but with ethyl 1 ,4- dioxaspiro[4.5]decane-7-carboxylate substituted for ethyl 1 ,4-dioxaspiro[4.5]decane-8- carboxylate.
Synthesis of 1.4-dioxaspiro[4.41nonane-7-carboxyiic acid
Figure imgf000389_0003
1 ,4-dioxaspiro[4.4]nonane-7-carboxylic acid was synthesized in a manner simiiar to the synthesis of 1 ,4-dioxaspiro[4.5]decane-8-carboxy!ic acid, but with ethyl 1 ,4- dioxaspiro[4.4]nonane-7-carboxylate substituted for ethyl 1 ,4-dioxaspiro[4.5]decane-8- carboxylate.
Synthesis of ethyl 3-oxo-3-(1.4-dioxaspiror4.51decan-7-yl)propanoate
Figure imgf000390_0001
Ethyl 3-oxo-3-(1 ,4-dioxaspiro[4.5]decan-7-yl)propanoate was synthesized in a manner similar to the synthesis of ethyl 3-oxo-3-(1 ,4-dioxaspiro[4.5]decan-8-yl)propanoate, but with 1 ,4-dioxaspiro[4.5]decane-7-carboxylic acid substituted for 1 ,4- dioxaspiro[4.5]decane-8-carboxylic acid.
Synthesis of ethyl 3-oxo-3-(1.4-dioxaspirof4.41nonan-7-yl)propanoate
Figure imgf000390_0002
Ethyl 3-oxo-3-(1 ,4-dioxaspiro[4.4]nonan-7-yi)propanoate was synthesized in a manner similar to the synthesis of ethyl 3~oxo-3-(1 ,4-dioxaspiro[4.5]decan-8-yi)propanoate, but with 1 ,4~dioxaspiro[4.4]nonane-7-carboxylic acid substituted for 1 ,4- dioxaspiro[4.5]decane-8-carboxylic acid.
Synthesis of 5-( 1.4-dioxaspirof4.5idecan-7-yl)pyrazolori ,5-aipyrimidin-7-ol
Figure imgf000390_0003
5-(1 ,4-dioxaspiro[4.5]decan-7-yl)pyrazolo[1 ,5-a]pyrimidin-7-o! was synthesized in a manner similar to the synthesis of 5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[ 1 ,5- a]pyrimidin-7-ol, but with ethyl 3-oxo-3-{1 ,4-dioxaspiro[4.5]decan-7-yl)propanoate substituted for ethyl 3-oxo-3-(1 ,4-dioxaspiro[4.5]decan-8-yl)propanoate. Synthesis of 5-(1 AdioxaspiroKΛInonan^-vQpyrazolori ^-alpyrimidin^-ol
Figure imgf000391_0001
5-(1 ,4-dioxaspiro[4.4]nonan-7-yl)pyrazolo[1 ,5-a]pyrimidin-7-o! was synthesized in a manner similar to the synthesis of 5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[1,5- a]pyrimidin-7-ol, but with ethyl 3-oxo-3-(1 ,4-dioxaspiro[4.4]nonan-7-yl)propanoate substituted for ethyl 3-oxo-3-(1 ,4-dioxaspiro[4.5]decan-8-y!)propanoate.
Synthesis of 7-chloro-5-(1 ,4-dioxaspiror4.51decan-7-y])pyrazolof1.5-aipyrimidine
Figure imgf000391_0002
7-Chloro-5-(1 ,4-dioxaspiro[4.5]decan-7-yl)pyrazo!o[1 ,5-a]pyrimidine is synthesized in a manner similar to the synthesis of 7-chloro-5-(1,4-dioxaspiro[4.5]decan-8- yl)pyrazolo[1 ,5-a]pyrimidine, but with 5-(1 ,4-dioxaspiro[4.5]decan-7-yl)pyrazolo[1 ,5- a]pyrimidin-7-ol substituted for 5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[1 ,5- a]pyrimidin-7-ol.
Synthesis of 7-chloro-5-( 1.4-dioxaspiroF4,41nonan-7-yl)pyrazoiof 1 ,5-aipyrimidine
Figure imgf000391_0003
7-Chioro-5-(1 ,4-dioxaspiro[4.4]nonan-7-y!)pyrazolo[1 ,5-a]pyrimidine is synthesized in a manner similar to the synthesis of 7-ch!oro-5-(1 ,4-dioxaspiro[4.5]decan-8- yl)pyrazolo[1 ,5-a]pyrimidine, but with 5-(1 ,4-dioxaspiro[4.4]nonan-7-yl)pyrazolo[1 ,5- a]pyrimidin-7-oi substituted for 5-{1 ,4-dioxaspiro[4.5]decan-8-y[)pyrazolo[1 ,5- a]pyrimidin-7-ol.
Synthesis of 5-(1 ,4-dioxaspirof4.5ldecan-7-yl)pyrazoiof1 ,5-a1pyrimidin-7-amine
Figure imgf000392_0001
5-(1 ,4-Dioxaspiro[4.5]decan-7-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine is synthesized in a manner similar to the synthesis of 5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[1 ,5- a]pyrimidin-7-amine, but with 7-chloro-5-(1 ,4-dioxaspiro[4.5]decan-7-yl)pyrazolo[1 ,5- a]pyrimidine substituted for 7-chloro-5-{1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazo!o[1 ,5- a]pyrimidine.
Synthesis of 5-(1.4-dioxaspiro[4.41nonan-7-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine
Figure imgf000392_0002
5-(1 ,4-Dioxaspiro[4.4]nonan-7-yl)pyrazo!o[1 ,5-a]pyrimidin-7-amine was synthesized in a manner similar to the synthesis of 5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazo!o[1 ,5- a]pyrimidin-7-amine, but with 7-chloro-5-(1 ,4-dioxaspiro[4.4]nonan-7~yl)pyrazolo[1 ,5- a]pyrimidine substituted for 7-chloro-5-(1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[1 ,5- a]pyrimidine. Synthesis of 3-(7-aminopyrazoiori ,5-aipyrimidin-5-yl)cyciohexanone
Figure imgf000393_0001
3-(7-Aminopyrazolo[1 ,5-a]pyrimidin-5-yl)cycIohexanone was synthesized in a manner similar to the synthesis of 4-(7-aminopyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexanone, but 5-{1 ,4-dioxaspiro[4.5]decan-7-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine substituted for 5- (1 ,4-dioxaspiro[4.5]decan-8-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine.
Synthesis of 3-(7-aminopyrazolo[1 ^-alpyrimidin-5-yl)cvclopentanone
Figure imgf000393_0002
3-(7-Aminopyrazolo[1 ,5-a]pyrimidin-5-yl)cyclopentanone was synthesized in a manner similar to the synthesis of 4-(7-aminopyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexanone, but 5-(1 ,4-dioxaspiro[4.4]nonan-7-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine substituted for 5- (1 Adioxaspiro[4.5]decan-8-yl)pyrazo]o[1 ,5-a]pyrimidin-7-amine.
Synthesis of 3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-aipyrimidin- 5-yl)cvciohexanone
Figure imgf000393_0003
3-(7-(Bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazo!o[1 ,5-a]pyrimidin-5- yl)cyclohexanone was synthesized in a manner similar to the synthesis of 4-{7-(bis((2- (trimethylsilylJethoxyJmethylJaminoJpyrazoloti .5-alpyrimidin-5-yl)cyclohexanone, but 3-(7-aminopyrazo!o[1 ,5-a]pyrimidin-5-yl)cyclohexanone substituted for 4-(7- aminopyrazo]o[1 ,5-a]pyrimidin-5-yl)cyclohexanone.
Synthesis of 3-{7-{ bis((2-(trimethylsilyl)ethoxy)methyi)amino)pyrazolof 1 ,5-aipyrimidin- 5-yl)cvclopentanone
Figure imgf000394_0001
3-(7-(Bis({2-{trimethylsilyl)ethoxy)methyt)amino)pyrazolo[1 ,5-a]pynmidin-5- yl)cyclopentanone was synthesized in a manner similar to the synthesis of 4-(7- (bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1,5-a]pyrimidin-5- yl)cyclohexanone, but 3-(7-aminopyrazolo[1 ,5-a]pyrimidin-5-yl)cyclopentanone substituted for 4-(7-aminopyrazolo[1 ,5-a]pyrimidin-5-yi)cyclohexanone.
Synthesis of ethyl 2-(3-(7-(bis(f2-(trimethylsilyl)ethoxy)methyl)amino)pyrazoio[1 T5- aipyrimidin-5-yl)cvclohexy!idene)acetate
Figure imgf000394_0002
Ethyl 2-(3-(7-(bis{(2-(trimethylsi!y!)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexylidene)acetate was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-{bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexylidene)acetate, but with 3-(7-(bis((2- (trimethylsilyl)ethoxyJmethyl)aminoJpyrazoloCi .5-alpyrimidin-5-ylJcyclohexanone substituted for 4-(7-(bis{(2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yi)cyclohexanone.
Synthesis of 2-(3-(7-(bis((2-(trimethylsilyl)ethoxy)methyπamino)pyrazolo[1 ,5- aipyrimidin-5-yl)cvclopentylidene)acetonitrile
Figure imgf000395_0001
2-(3-(7-(bis((2-{trimethylsiiy!)ethoxy)πnethyl)amino)pyrazolo[1 )5-a]pyrimidin-5- yl)cyclopentylidene)acetonitriie was synthesized in a manner similar to the synthesis of 2-(4-(7-(Bis({2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1,5-a]pyrimidin-5- yi)cyclohexylidene)acetonitrile, but with 3-(7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyhmidin-5-yl)cyclopentanone substituted for 4-{7-(bis((2-(trimethylsilyl)ethoxy)methyS)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexanone.
Synthesis of ethyl 2-(3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazoloM ,5- aipyri m idi n-5-yl )cycioh exyl) acetate
Figure imgf000395_0002
Ethyl 2-(3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate was synthesized in a manner similar to the synthesis of ethyl 2- {4-{7-(bis((2-(trimethylsiiy!)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyciohexyl)acetate, but with ethyl 2-(3-(7-(bis((2- (trimethylsilyl)ethoxyJmethyl)aminoJpyrazoloti .5-alpyrimidin-5- yl)cyclohexyliclene)acetate substituted for ethyl 2-(4-(7-(bis{(2- (trimethylsilyI)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexylidene)acetate.
Synthesis of 2-(3-(7-(bisf(2-(trimethylsiiyl)ethoxy)methyl)amino)pyrazolo[1 ,5- alpyrimidin-5-yl)cvclopentyl)acetonitrile
Figure imgf000396_0001
2-(3-(7-(Bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 I5-a]pyrimidin-5- yl)cyciopentyl)acetonitrile was synthesized in a manner similar to the synthesis of ethyl 2-{4-(7-(bis((2-{trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 I5-a]pyrimidin-5- yl)cyclohexyl)acetate, but 2-(3-(7-(bis((2- {trimethylsiiyi)ethoxy)methyl)amino)pyrazolo[1 ,5-a3pyrimidin-5- yl)cyclopentylidene)acetonitrile substituted for ethyl 2-(4-(7-(bis((2- (trimethy!silyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexylidene)acetate.
Synthesis of ethyl 2-(3-(7-(bisff2-(thmethylsilvhethoxy)methyπaminoV3- iodopyrazoloπ .5-alpyrimidin-5-yl)cvciohexyl)acetate
Figure imgf000396_0002
Ethyl 2-(3-(7-(bis((2-(trimethylsiiyl)ethoxy)methy])amino)-3-iodopyrazoto[1 ,5- a]pyrimidin-5-yl)cyclohexyi)acetate was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsiiyi)ethoxy)methyl)amino)-3- iodopyrazoio[1 ,5-a]pyrimidin-5-yl)cyciohexy!)acetate, but with 2-{3-{7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclopentyl)acetonitrile substituted for ethyl 2-(4-(7-(bis((2- (trimethylsilyiJethoxyJmethylJaminoJpyrazoloEI .5-alpyrimidin-5-yl)cyclohexyl)acetate.
Synthesis of 2-(3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazoio[1 ,5- alpyrimidin-5-yl)cyclopentvPacetonitrile
Figure imgf000397_0001
2-{3-(7-(Bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclopentyl)acetonitrile was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis((2-(trimeihylsilyi)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyi)acetate, but with ethyl 2-(3-(7-(bis({2- (tnmethylsilyl)ethoxyJmethylJaminoJpyrazolofi .5-aJpyrimidin-5-yl)cyclohexylJacetate substituted for ethyl 2-(4-(7-{bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexyl)acetate.
Synthesis of ethyl 2-(3-(7-(bis((2-(trimethy[silyl)ethoxy)methy[)amino)-3-(quinoiin-3- yl)pyrazolori .5-aipyrimidin-5-vOcvclohexyl)acetate
Figure imgf000398_0001
Ethyl 2-(3-(7-(bis((2-(trimethylsilyI)ethoxy)methyl)amino)-3-(quinolin-3-yl)pyrazolo[1 l5- atøyrimidin-5-yl)cyclohexyl)acetate was synthesized in a manner similar to the synthesis of ethyl 2-(4-{7-{bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quino!in-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-y!)cyclohexyi)acetate, but with ethyl 2-(3-(7-(bis({2- (trimethylsilyI)ethoxy)methy!)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate substituted for ethyl 2-(4-(7-(bis((2- (trimethylsilyi)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate.
Synthesis of 2-(3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolori ,5-aipγrimidin-5-yl)cvclopentyl)acetonitrile
Figure imgf000398_0002
2-(3-(7-(Bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclopentyl)acetonitrile was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1.5-aføyrimidin-5-ylJcyclohexyl)acetate, but with 2-(3-(7-(bis((2- (trimethyJsilyl)ethoxy)methyl)amino)-3-iodopyrazo[o[1,5-a]pyrimidιn-5- yl)cyclopentyl)acetonitrile substituted for ethyl 2-(4-(7-(bis((2-
(trimethylsiiyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate. Synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(1-methyl-1 H- pyrazol^-yl)pyrazoloH .5-alpyrimidin-5-yl)cvclohexyl)acetate
Figure imgf000399_0001
Ethyl 2-(4-{7-(bis((2-(trimethylsilyI)ethoxy)methyl)amino)-3-(1-methyl-1 H-pyrazol-4- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetate was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis{(2-(trimethylsilyl)ethoxy)methyl)amino)-3- (quinolin-3-yl)pyrazo!o[1 ,5-a3pyrimidin-5-y!)cyclohexyi)acetate, but with N-methyl pyrazo!e-4-boronic acid pinacol ester substituted for quinoline-3-boronic acid.
Synthesis of 2-(3-(7-amino-6-bromo-3-(αuinolin-3-yl)pyrazolo[1 ,5-alpyrimidin-5- vOcyclohexyltecetic acid
Figure imgf000399_0002
2-(3-(7-Amino-6-bromo-3-(quinolin-3-y!)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetic acid was synthesized in a manner similar to the synthesis of 2-(4-(7-amino-6-bromo-3- (quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetic acid, but with ethyl 2-(3- {7-{bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinotin-3-yl)pyrazolo[1 t5- a]pyrimidin-5-yl)cyclohexyl)acetate substituted for ethyl 2-(4-(7-(bis((2- {trimethylsiiy!)ethoxy)methyl}amino)-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate. The reaction was concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield 2-(4-(7-amino-6-bromo-3-(quinolin~3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexy[)acetonitrile as a four-isomer mixture. (m+H = 480.1 , retention time = 3.51 min)
Synthesis of 2-(3-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolori ,5-a1pyrimidin-5- yl)cvclopentyl )aceton it ri Ie
Figure imgf000400_0001
2-(3-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- y!)cyclopentyl)acetonitrile was synthesized in a manner similar to the synthesis of 2- (4-(7-amino-6-bromo-3-(quinolin-3-yI)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidin-1 -yl)acetic acid, but with 2-{3-(7-(bis((2-(trimethylsiIyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1.5-alpyrimidin-5-ylJcyclopentyl)acetonitrile substituted for tert-butyl 2-(4- (7-amino-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidin-1 -yljacetate. The reaction was concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield 2-(3-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclopentyl)acetonitrile as a yellow solid. (m+H = 447.22, retention time = 3.99 min).
Figure imgf000401_0001
SCHEME- 6
Synthesis of tert-butyl 4-(7-hydroxypyrazolori ,5-alpyrimidin-5-yl)piperidine-1- carboxylate
Figure imgf000401_0002
tø/t-Butyl 4-(7-hydroxypyrazoio[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxylate was synthesized in a manner similar to the synthesis of 5-(1 ,4-dioxaspiro[4,5]decan-8- yl)pyrazolo[ 1 ,5-a]pyrimidin-7-ol, but with tert-butyl 4-(3-ethoxy-3- oxopropanoyl)piperidine-1 -carboxylate substituted for ethyl 3-oxo-3-(1 ,4- dioxaspiro[4.5]decan-8-yl)propanoate.
Synthesis of tert-butyl 4-(7-chloropyrazolori .5-alpyrimidin-5-vnpiperidine-1- carboxviate
Figure imgf000402_0001
tert-Butyl 4-(7-chloropyrazo!o[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxylate was synthesized in a manner similar to the synthesis of 7-chloro-5-(1 ,4- dioxaspiro[4.5]decan-8-yI)pyrazolo[1.5-ajpyrimidine, but with te/t-butyl 4-(7- hydroxypyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxylate substituted for 5-(1 ,4- dioxaspiro[4.5]decan-8-yl)pyrazolo[1 ,5-a]pyrimidin-7-oi.
Synthesis of fert-butyl 4-(7-aminopyrazoloM ,5-aipyrimidin-5-γl)piperidine-1- carboxylate
Figure imgf000402_0002
tert-Butyl 4-(7-aminopyrazolo[1 ,5-a]pyhmidin-5-yl)piperidine-1 -carboxylate was synthesized in a manner similar to the synthesis of 5-{1 ,4-dioxaspiro[4.5]decan-8- yl)pyrazolo[1 ,5-a]pyrimidin-7-amtne, but tert-butyl 4-(7-chloropyrazo!o[1 ,5-a]pyrimidin- 5-y[)piperidine-1 -carboxylate substituted for 7-ch!oro-5-(1 ,4-dioxaspiro[4.5]decan-8- yi)pyrazolo[1 ,5-a]pyrimidine.
Synthesis of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazoloF1 ,5- aipyrimidin-5-yl)pipeπdine-1 -carboxylate
Figure imgf000402_0003
tert-Butyl 4-(7-(bis{(2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)piperidine-1-carboxylate was synthesized in a manner similar to the synthesis of 4- (7-(bis((2-(trimethylsilyl)ethoxy)methyl}amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexanone, but with te/t-butyl 4-(7-aminopyrazolo[1 ,5-a]pyrimidin-5- yi)piperidine-1-carboxylate substituted for 4-(7-aminopyrazolo[1 ,5-a]pyrimidin-5- yl)cyciohexanone.
Synthesis of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3- iodopyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxyiate
Figure imgf000403_0001
tert-Buty\ 4-(7-(bis((2-(trimethylsiiyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5- a]pyrimidin-5-y[)piperidine-1 -carboxyiate was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3- iodopyrazoioπ^-ajpyrimidin-5-ylJcyclohexyl)acetate, but with tørf-butyl 4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amιno)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 - carboxylate substituted for ethyl 2-(4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexyl)acetate.
Synthesis of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1 ,5-aipyrimidin-5-yl)piperidine-1-carboxvlate
Figure imgf000403_0002
tert-Butyl 4-(7-(bis({2-(thmethylsilyl)ethoxy)methyl)amino)-3-(quinoiin-3- y!)pyrazo!o[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxyiate was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis({2-(trimethylsilyl)ethoxy)methyl)amino)-3- (quinolin-3-yi)pyrazolo[1.5-alpyrimidin-5-yl)cyclohexylJacetate, but with førf-butyl 4-(7- (bis((2-(trimethylsilyi)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)piperidine-1-carboxylate substituted for ethyl 2-{4-(7-(bis((2- {trimethylsilyl)ethoxy)methyl)amino)-3~iodopyrazolo[1 ,5-ajpyrimidin-5- yl)cyclohexyl)acetate.
Synthesis of te/f-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)aminoV3-(1-methyl- 1 H-pyrazol-4-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)piperidine-1 -carboxyiate
Figure imgf000404_0001
te/t-Butyl 4-(7-(bis({2-{trimethylsilyl)ethoxy)methyl)amino)-3-(1 -methyl- 1 H-pyrazoi-4- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxyiate was synthesized in a manner similar to the synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsily!)ethoxy)methyl)amino)-3- {1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1.5-alpyrimidin-5-yl)cyclohexyl)acetate, but with tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5- a]pyrimidin-5-yi)piperidine-1 -carboxyiate substituted for ethyl 2-(4-(7-(bis{(2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate.
Synthesis of 5-(piperidin-4-vO-3-(cιuino[in-3-yl)pyrazolori ,5-aipyrimidin-7-amine
Figure imgf000404_0002
To a 20 mL scintillation vial was charged tert-butyl 4-(7-{bis((2- (trimethylsilyl)ethoxy)methyl)-amino)-3-(quinol!n-3-yl)pyrazoio[1 ,5-a]pyrimidin-5- yl)piperidine-1 -carboxylate (80 μmol, 56 mg) followed by ethanol (2 mL) and H2O (500 μ[_). To this solution was added 2 mL of 4N HCI in 1 ,4-dioxane. This solution was heated to 60° C for 2 hours. Upon completion, the reaction mixture was diluted with DCM (10 mL) and washed with NaHCO3(aq) and extracted with DCM twice more. The combined organics were then dried over Na2SO4 and the solvent was removed in vacuo to yield 5-(piperidin-4-yl)-3-(quinolin-3-yl)pyrazoio[1 ,5-a]pyrimidin-7-amine (20.3 mg, 59 umoi) as cream-colored solid .
Synthesis of tert-butyl 2-(4-(7-amino-3-(quinolin-3-yl)pyrazolori ,5-a1pyrimidin-5- yl)piperidin-1 -yl)acetate
Figure imgf000405_0001
To a 20 mL scintillation vial was charged tert-butyl 2-(4-(7-amino-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyrirnidin-5-yl)piperidin-1-yl)acetatec (100 μmol, 34 mg) followed by a solution of N,N-diisopropylethylamine (150 μmol, 26 μL) in DMF (2 mL). To the resulting solution was added te/t-butyl 2-bromoacetate (120 μmol, 18 μL). The reaction mixture was stirred at 60° C for 18 hours. After 18 hours, the reaction mixture was concentrated in vacuo and purified via silica gel chromatography (0% to 20% MeOH in DCM) to yield ført-butyl 2-(4-(7-amino-3-(quinolin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)piperidin-1 -y[)acetate (41 mg, 90 μmol).
Synthesis of 2~(4-(7-amino-6-bromo-3-(quinolin~3-yl)pyrazolof 1 ^-alpyrimidin-5- yl)piperidin-1-yl)acetic acid
Figure imgf000406_0001
To a 20 mL scintillation vial was charged 2-(4-(7-amino-6-bromo-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperic!in-1-yl)acetic acid (41 mg, 90 μmol) plus acetonitrile (2 mL). To this solution was added N-bromosuccinimide {90 μmol, 16 mg). This solutionwas allowed to stir at room temperature for 1 hour. The reaction was concentrated in vacuo and dissolved in TFA (2 mL). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield 2-(4-(7-amino-6-bromo-3- (quino!in-3-yl)pyrazolo[1T5-a]pyrimidin-5-yi)piperidin-1-yl)acetic acid as yellow solid. (m+H = 481.22, retention time = 2.32 min)
Synthesis of 3-(1 -methyl-1 H-pyrazol-4-yl)-5-(piperidin-4-vQpyrazoloπ ,5-a1pyrimidin-7- amine
Figure imgf000406_0002
3-(1 -Methyl-1 H-pyrazol-4-yl)-5-(piperidin-4-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine was synthesized in a manner similar to the synthesis of 5-(piperidin-4-yl)-3-(quinolin-3- yl)pyrazoio[1 ,5-a]pyrimidin-7-amine, but with tert-butyl 4-(7-(bis((2- (trimethylsilyi)ethoxy)methyl)amino)-3-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5- a]pyrimidin-5-yi)piperidine-1-carboxylate substituted for tert-butyl 4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-(quino!in-3-yl)pyrazolo[1 ,5-a3pyrimidin-5- yl)piperidine-1 -carboxylate. Synthesis of tert-butyl 2-(4-(7-amino-3-(1 -methyl-1 H-pyrazol-4-yl)pyrazolori ,5- aipyrimidin-5-yl)piperidin-1-yl)acetate
Figure imgf000407_0001
te/t-Butyl 2-(4-(7-amino-3-(1-methyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)piperidin-1-yl)acetate was synthesized in a manner similar to the synthesis of tert- butyl 2-(4-(7-amino-3-(quino!in-3-yl)pyrazoio[1 ,5-a]pyrimidin-5-yl)piperidin-1 - yl)acetate, but with 3-(1 -methyl-1 H-pyrazo!-4-yl)-5-(ptperidin-4-yl)pyrazolo[1 ,5- a]pyrimidin-7-amine substituted for 5-(piperidin-4-yi)-3-(quinolin-3-yl)pyrazoIo[1 ,5- a]pyrimidin-7-amine.
Synthesis of 2-(4-(7-amino-6-bromo-3-( 1-methyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5- alpyrimidin-5-vOpiperidin-i -ypacetic acid
Figure imgf000407_0002
2-(4-{7-amino-6-bromo-3-(1 -methyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)piperidin-1-yl)acetic acid was synthesized in a manner similar to the synthesis of 2- (4-(7-amino-6-bromo-3-(quinolin-3-yJ)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidin-1-yl)acetic acid, but with tert-butyl 2-(4-(7-amino-3-(1-methyI-1 H-pyrazol-4-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)piperidin-1-yl)acetate substituted for tert-butyl 2-{4-(7-amino-3- (quinolin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidin-1-yl)acetate. The reaction mixture wass reduced in vacuo and purified via reverse-phase preparatory HPLC to yieid 2-(4-(7-amino-6-bromo-3-(1-methyl-1 H-pyrazol-4-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)piperidin-1-yl)acetic acid as off-yellow solid. (m+H = 434.19, retention time = 2.25 min)
tert-butyl 3-(7-hydroxypyrazolof 1 ,5-a1pyrimidin-5-yl)pyrrolidine-1 -carboxylate
Figure imgf000408_0001
A mixture of tert-butyl 3-(3-ethoxy-3-oxopropanoyl)pyrrolidine-1 -carboxylate (7.00 g, 25 mmol) and 1 H-pyrazol-3-amine (2.08 g, 25 mmol) in toluene 30 mL) was heated at 1 10 °C under argon for 18 h and concentrated in vacuo. The residue was triturated with EtOAc and the solid was collected by filtration and dried in high vacuum to give 6.84 g (90%) of the title compound as a white solid. LC/MS RT = 1.40 min (5 min method). Mass calculated for, M+H 305.15, observed 305.15.
tert-butyl 3-(7-chloropyrazolo|"1 ,5-a1pyrimidin-5-yl)pyrrolidine-1 -carboxylate
Figure imgf000408_0002
To a mixture of tert-butyl 3-(7-hydroxypyrazoio [1 , 5-a] pyrimidin-5-yl) pyrrolidine-1 - carboxylate (3.70 g, 12.0 mmol) and dimethylaniline (4.0 mL, 30 mml) was added phosphoryl trichloride (40 mL). The mixture was heated at 50 °C for 5 h and concentrated in vacuo. The residue was diluted with methylene chloride (100 mL) and quenched with saturated NaHCOe (100 mL). The mixture was separated and the aqueous layer was extracted with chloride (100 mL x Z). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by Biotage (CH2Ci2/Et0Ac, 20:1 to 4:1). LC/MS RT = 2.09 min (5 min method). Mass calculated for, M+H 322.12, observed 337.14. tert-butyl 3-( 7-aminopyrazolo[1 ,5-a1pyrimidin-5-yl)pyrroiidine-1 -carboxyiate
Figure imgf000409_0001
A mixture of tert-butyl 3-(7-chloropyrazolo [1 , 5-a] pyrimidin-5-yl) pyrroiidine-1- carboxyiate (3.40 g, 10.5 mmol) in a solution of ammonia in MeOH (7N1 20 mL) was heated in sealed vessel at 80 °C for 5 h. The reaction mixture was cooled down and concentrated in vacuo. The residue was dried in high vacuum and used for the next step without further purification. LC/MS RT = 1.30 min (5 min method). Mass calculated for, M+H 303.17, observed 318.19.
tert-butyl 3-(7-(bisf(2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-aipyrimidin-5- vQpyrrolidine-1 -carboxviate
Figure imgf000409_0002
To a mixture of tert-butyl 3-(7-aminopyrazolo [1 , 5-a]pyrimidin-5-yi)pyrrolidine-1 - carboxyiate (10.5 mmol) in CH2CI2 was added DIEA (10.8 mL, 63 mmol) and SEM-CI (5.6 mL, 32 mmol). The mixture was heated at 50 °C for 5 h under an argon atmosphere. The mixture was cooled, and the reaction was quenched with saturated NaHCO3 (100 mL) and extracted with CH2CI2 (50 mL x 3). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by Biotage (CH2CI2/EtOAc, 20:1 to 5:1 ) to give the title compound as a light brown oil. LC/MS RT - 2.90 min (5 min method). Mass calculated for, M+H 563.33, observed 578.35.
tert-butyl 3-(7-(bisf(2-(trimethylsiiyl)ethoxy)methyl)amino)-3-iodopyrazolori .5- aiPvrimidin-5-vhpvrroiidine-1 -carboxviate
Figure imgf000410_0001
To a solution of tert-butyl 3-(7-(bis((2-(trimethylsilyl)ethoxy)methy!)amino)pyrazoio[1 ,5- a]pyrimidin-5-yl)pyrroliciine-1-carboxylate 3.95 g, 7.0 mmol) in CH3CN {30 mL) was added NIS {1.65 mg, 7.0 mmol). The mixture was stirred at rt for 2 h and diluted with EtOAC (50 mL) and washed with saturated sodium thiosulfate, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by Biotage (CH2CI2/Et0Ac, 100:1 to 10:1 ) to the title compound as a light brown oil. LC/MS RT = 2.49 min. Mass calculated for, M+H 689.23, observed 389.17.
tert-butyl 3-(7-(bisf(2-(trimethylsilyl)ethoxy)methyhamino)-3-(quinolin-3- yhpyrazoioπ .5-alpvrimidin-5-vhpyrrolidine-1 -carboxylate
Figure imgf000410_0002
to 5 h N(SEM)2
A mixture of tert-butyl 3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3- iodopyrazolo[1 ,5-a]pyrirnidin-5-yl)pyrrolidine-1 -carboxylate (703 moi), quinolin-3- ylboronic acid (350 mg, 2.00 mmol), Pd(dppf)CI2-CH2CI2 (82 mg, 0.10 mmol) and potassium phosphate (636 mg, 3.0 mmol) in a mixture of dioxane/H2O (9:1 , 20 mL) was heated at 100 0C for 5 h under argon. The reaction mixture was cooled down and diluted with EtOAC, washed with Brine, dried over Na2SO4, filtered and concentrated. The residue was purified by Biotage (CH2CI2ZEtOAc, 10:1 to 3:1 ) to give the title compound as brown solid. LC/MS RT = 2.49 min. Mass calculated for, M+H 690.37, observed 389.17.
tert-butyl 3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(αuinoiin-3- yl)pyrazoloπ ,5-aipyrimidin-5-yl)piperidine-1 -carboxyiate
Figure imgf000411_0001
To a solution of tert-butyl 3-{7-(bis((2-(trimethylsiIyl)ethoxy)methyl)amino)-3-(quinolin- 3-y!)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxyiate (350 mg, 0.50 mmol) in CH3CN (5.0 mL) was added NBS (90 mg, 0.50 mmol). The mixture was stirred at rt for 3 h and concentrated in vacuo. The residue was purified by Biotage (CH2CI2/Et0Ac, 100:1 to 10:1) to give the title compound as light yellow solid. LC/MS RT = 2.49 min. Mass calculated for, M+H 768.29, observed 389.17.
6-bromo-5-(piperidin-3-yl)-3-(quinoiin-3-yl)pyrazolo[1 ,5-aiPyrimidin-7-amine
Figure imgf000411_0002
To a solution of tert-butyl 3-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromσ-3- (quinolin-3-yl)pyrazoio[1 ,5-a3pyrimidin-5-yl)piperidine-1-carboxylate (196 mg, 0.40 mmol) in EtOH (5.0 mL) was added 4 N HCI (5.0 mL). The mixture was heated at 60 °C for 4 h under argon, cooled down and concentrated in vacuo. The residue was dried in high vacuum to give the title compound as yellow solid (HCI salt). LC/MS RT = 2.49 min. Mass calculated for, M+H 408.07, observed 389.17.
6-bromo-5-(1 -(methylsulfonyl)piperidin-3-yl)-3-(quinolin-3-yl)pyrazolo[1 ,5-aipyrimidin- 7-amine
Figure imgf000411_0003
To a mixture of 6-bromo-5-(piperidin-3-y!)-3-(quinoIin-3-yi)pyrazolo[1 ,5-a]pyrimidin-7- amine (HCI salt, 46 mg, 0.10 mmol) in DMF (2.0 ml_) was added DIEA (0.10 mL, 0.30 mmol) and methylsuifonyl chloride (14 mg, 0.12 mmol). The reaction mixture was stirred at rt for 3 h and concentrated in vacuo. The residue was purified by prep-LC to give the title compound. LC/MS RT = 2.49 min. Mass calculated for, M+H 486.05, observed 389.17.
By essentially the same procedure given in Schemes 25, the compounds listed in Table 8 can be prepared.
Table 8
Figure imgf000412_0001
Figure imgf000413_0001
Figure imgf000414_0001
Figure imgf000415_0001
Alternative to the described procedure in the Scheme-14, these compounds can aiso be made by the scheme 25 given below. 7-Amino-3-(6-Dhenylpyridin-3-yl)pyrazoloπ .5-aipyrimidin-5-yl)piperidine-1 -carboxyiate:
Figure imgf000416_0001
SCHEME-26
Step-1
Synthesis of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amSno)pyrazoioπ ,5- alpyrimidin-5-yl)-5,6-dihydropyridine-1 (2H)-carboxylate
Figure imgf000416_0002
DME, 2M Na2CO3
100 °C
To a pressure tube were charged 5-chloro-N,N-bis((2-
(trimethylsϋylJethoxyJmethyl)pyrazoloII ^-alpyrimidin-y-amine {5.36 g, 12.5 mmols), tert-butyl 4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaboroian-2-yl)-5,6-dihydropyridine-1 (2H)- carboxylate (4.26 g, 13.8 mmols), PdC!2(dppf).CH2Cl2 ([1 ,1 '- bis(diphenylphosphino)ferrocene]dichloropal!adium(il) complex with dichloromethane (1 :1 ) ) (510 mg, 0.62 mmol), 2M Na2CO3 (30 mL) and DME (60 ml_). The tube was degassed with Ar briefly, capped and heated at 100 0C with stirring overnight. After cooling, the reaction mixture was diluted with EtOAc and water, organic layer was isolated, washed with brine and dried (MgSO4). After solvent was removed under reduced pressure, the residue was purified on silica. Elution with EtOAc in hexanes (0-25%) gave tert-butyl 4-(7-(bis((2-(trimethylsilyi)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)-5,6-dihydropyridine-1(2H)-carboxylate (5.76 g, 80%).
Step 2
Synthesis of fe/t-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolof 1 ,5- aipyrimidin-5-yl)piperidine-1-carboxyiate
Figure imgf000417_0001
A mixture of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)-5,6-dihydropyridine-1 (2H)-carboxylate (5.05 g,8.78 mmoi) and 10% Pd/C (100 mg) in EtOAc was stirred at 45 °C under hydrogen (balloon pressure) for three hours. After filtration, washing with EtOAc (3x) and concentration, te/t-butyl A- (7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine- 1 -carboxyiate (5.1 g, 99.5%) was obtained as oil.
Step 3 Synthesis of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyhamino)-3- iodopyrazolori ,5-aipyrimidin-5-yl)piperidine-1-carboxviate
Figure imgf000417_0002
To a mixture of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5- a]pyrimidin-5-yl)piperidine-1 -carboxyiate (5 g, 8.65 mmol) in DMF (15 ml_) was added N-iodosuccinimide (1.86 g, 8.27 mmoi) and the resulting mixture was allowed to stir at room temperature for 1 hour. The mixture diluted with EtOAc (100 ml_), washed with water (20 mL). The aqueous layer was extracted with EtOAc (20 ml_) once, and combined organic layer was washed with water (20 mL) three times, brine once and dried (MgSO4). After concentration, the residue was purified by column chromatography on silica gel. Elution with EtOAc/Hex (0-30%) gave tert-butyl 4-(7- (bis((2-(tπmethylsilyl)ethoxy)methyl)amino)-3-iodopyrazoio[1 ,5-a]pyrimidin-5- yl)piperidine-1 -carboxyiate (4.55 g, 75%). (Note: DMF could be replaced by acetonitrile.)
Step 4 Synthesis of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyQamino)-3-(6- phenv[pyridin-3-yl)pyrazoloM,5-aiPyrimidin-5-yl)piperidine-1-carboxylate
DME/water
Figure imgf000418_0001
To a pressure tube were charged te/ϊ-butyl 4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 - carboxylate (4.2 g, 5.97 mmol), 2-phenyl-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)pyridine (2 g, 7.1 mmol), PdCI2(dppf).CH2CI2 {240 mg, 0.33 mmol), DME (16 ml_) and 2M Na2Cθ3 (8 mL). The mixture was briefly degassed with Argon and the tube was capped and heated at 100 °C for 15 hours. On cooling, H2O (20 mL) and EtOAc (40 mL) and aqueous layer was extracted with EtOAc (3x) and combined organic layers were washed with brine once and dried (MgSO4). After concentration in vacuo the residue was purified on silica gel. Elution with EtOAc/Hexanes (0-40%) gave the title compound (3.24 g, 74%).
Step 5 Preparation of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyDamino)-6-bromo-3- (6-phenylpyridin-3-yl)pyrazolori,5-a1pyrimidin-5-yl)piperidine-1 -carboxvlate
Figure imgf000418_0002
To a solution of tert-buty! 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)pyrazo!o[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxylate (3.24 g, 4.44 mmo!) in DMF (20 mL) was added NBS (750 mg, 4.21 mmol). After stirring at rt for one hour, NBS (75 mg 0.42 mmol) was added and the resulting mixture was stirred for another 40 minutes to complete the reaction. The reaction mixture was diluted with EtOAc (200 mL), washed with water (2x50 mL), brine (20 mL) and dried (MgSO4). After concentration in vacuo the residue was purified on silica gel. Elution with EtOAc/Hexanes (0-40%) gave the title compound (3 g, 83%).
Step-6 General method:
Figure imgf000419_0001
To a solution of the piperidine derivative (1 eq), DlEA (4 eq) in DMF was added appropriate coupling reagent (1 eq): chloroformate to form carbamate; isocyanate to provide urea; suifonyl chloride to form sulfonamide; and carboxylic acid/EDCI/HOBt to yield the corresponding amide. The reaction was monitored by HPLC. Once the reaction was complete, the reaction mixture was directly purified by HPLC to furnish the corresponding product.
TABLE-8A
Figure imgf000420_0001
Figure imgf000421_0001
Figure imgf000422_0001
Figure imgf000423_0001
Figure imgf000424_0001
Figure imgf000425_0001
Figure imgf000426_0001
Figure imgf000427_0001
Figure imgf000428_0001
Figure imgf000429_0001
Figure imgf000430_0001
Figure imgf000431_0001
Figure imgf000432_0001
Figure imgf000433_0001
Synthesis of (RH-(4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolori ,5- a1pyrimidin-5-yl)piperidin-1 -yl)-2-hydroxypropan-1 -one
Figure imgf000434_0001
Step 6
Preparation of tert-butyl 4-(7-(bisf(2-(trimethylsilyl)ethoxy)methyπamino)-6-(1 - ethoxyylnyl)-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)piperidine-1- carboxylate
Figure imgf000434_0002
To a pressure flask were charged compound fert-buty! 4-(7-(bis((2- (trimethylsilyl}ethoxy)methy!)amino)-6-bromo-3-{6-phenylpyridin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)piperidine-1 -carboxylate ( intermediate from Scheme-10A) (13.66 g, 16.86 mmol), tributyl (i-ethoxyvinyl)tin (11.4 mL, 33.74 mmols) and dioxane (150 ml). The flask was degassed with Ar briefly, capped and heated at 100 C with stirring overnight. After cooling, the reaction mixture was concentrated. The residue was taken into ether (200 ml), washed with 0.5 M KF (50 ml), brine and dried (MgSO4). After solvent was removed under reduced pressure, the residue was purified on silica. Elution with EtOAc in hexanes (0-30%) gave compound tert-butyl 4- (7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-{1 -ethoxyvinyl)-3-(6-phenylpyridin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1-carboxylate (1 1.55 g, 85%) as yellow form.
Step 7
Preparation of 1 -(7-amino-3-(6-phenylpyridin-3-yl)-5-(piperidin-4-yl)pyrazoiori ,5- aipyhmidin-6-yl)ethanoπe
Figure imgf000435_0001
To a solution of tert-butyl 4-(7-(bis({2-(trimethylsilyl)ethoxy)methyl)amino)-6-(1- ethoxyvinyl)-3-(6-phenylpyridin-3-y!)pyrazoio[1 ,5-a]pyrimidin-5-yi)piperidine-1- carboxyiate (1 1.5 g, 14.4 mmol) in dioxane (60 mL) was added 4M HCI in water (14.4 mL) at 0 °C. After stirring for 30 min at 0 °C, the reaction mixture was allowed to warm to rt in 1.5 hours. 4M HCl in dioxane (10 mL) was added. The reaction mixture was stirred at rt overnight. Solvent was removed in vacuo to get the desired product as an HCI salt (6.97 g).
Step 8
Preparation of (R)-1-(4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolof1 ,5- alpyhmidin-5-γ[)piperidin-1 -yl)-2-hydroxypropan-1 -one
Figure imgf000435_0002
To a solution of 1-(7-amino-3-(6-phenylpyridin-3-yl)-5-(piperidin-4-yl)pyrazoio[1 ,5- a]pyrimidin-6-yl)ethanone hydrochloride (0.5 mmol), DIEA (350 μL, 2 mmol) in DMF (5 mL) was added a solution of D-lactic acid (45 mg, 0.5 mmol), HOBt (68 mg, 0.5 mmol), EDCI (144 mg, 0.75 mmol) in DMF (1 mL). The reaction was stirred at rt for 20 min. Another solution of solution of D-lactic acid (45 mg, 0.5 mmol), HOBt (68 mg, 0.5 mmol), EDCI (144 mg, 0.75 mmol) in DMF (2 mL) was added and stirred for 5 minutes. LC-MS indicated the reaction was complete and reaction mixture was directly purified by HPLC to furnish the title compound.
LC/MS RT = 3.22 min. Mass calculated for M+H 485.2, observed 485.4.
Synthesis of (R)-1-(4-(3-(6-( 1H-1 , 2,4~triazol-1-vnpyridin-3-ylV-6-acetyl-7- aminopyrazoioh ,5-alpyrimidin-5-yl)piperidin-1 -yl)-2-hydroxypropan-1 -one
Figure imgf000436_0001
Step 1
Preparation of 5-bromo-2-(1 H-1 ,2,4-triazol-1 -yl)pyridine:
Figure imgf000436_0002
A mixture of 1 ,2,4-triazole {1.17 g, 16.9 mmol), 2,5-dibromopyridine (2 g, 8.4 mmol) and K2CO3 (1.2 g, 8.7 mmol) in NMP (20 mL) was heated at 100 °C with stirring for 5 hours. After cooling, the reaction mixture was poured into water (100 mL) and solid was collected by filtration and further purified by column chromatography on silica gel. Elution with EtOAc/Hexanes (0-50%) provided the title compound (250 mg, 13%).
Step 2
Preparation of 5-(4.4.5.5-tetramethyl-1 ,3,2-dioxaborolan-2-ylV-2-(1 H-1 ,2,4-triazol-1- vppyridine:
Figure imgf000437_0001
A mixture of 5-bromo-2-(1 H-1 ,2,4-triazol-1 -yl)pyridine (248 mg, 1.1 mmol), bis(pinacolato)diboron {337 mg, 1.4 mmol), KOAc (324 mg, 3.3 mmol), PdCI2(dppf)2.CH2CI2 (45 mg, 0.06 mmol) in dioxane (4 mL) was flushed with Argon and stirred at 80 °C for 16h. On cooling, the solvent was rotoevaporated, and the crude was redissolved in dichloromethane (20 mL), washed with water (3 x 10 ml), brine (1 x 101 ml), and dried over MgSO4. Solvent was removed in vacuo and the residue was triturated with hexanes to provide the title compound (287 mg, 96%).
Step 3
Preparation of fert-butyl 4-(3-(6-(1 H-1.2.4-triazol-1-vnpyridin-3-yl)-7-(bisff2-
(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]Dyrimidin-5-yl)piperidine-1 - carboxyiate:
Figure imgf000437_0002
To a pressure tube were charged tert-butyl 4-(7-(bis((2- (trimethy!silyl)ethoxy)methyl)amino)-3-iodopyrazoio[1 ,5-a]pyrimidin-5-yl)piperidine-1 - carboxylate (563 mg, 0.8 mmoi), 5-{4,4,5,5-tetramethyl-1 ,3,2-dioxaboroian-2-yl)-2- (1 H-1 ,2,4-triazol-1-yl)pyridine (287 mg, 1.1 mmol), PdCI2JdPPf)2-CH2CI2 (29 mg, 0.04 mmol), DME (6 mL) and 2M Na2CO3 (2 mL). The resulting mixture was briefly degassed with Argon; the tube was capped, and heated with stirring under 80 ° C overnight. After cooling, solvent was removed. The residue was diluted with EtOAc (20 ml), and organic layer was isolated, washed with brine and dried (MgSO4). After concentration under reduced pressure, the residue was purified on silica gel eluting with EtOAc/Hexanes (0-50%) to provide the title compound (370 mg, 64%). Step 4
Synthesis of (RH -(4-(3-(6-(1 H-1.2.4-triazol-1-vθDyridin-3-yl)-6-acetyl-7- aminopyrazolof 1 ,5-a1pyrimidin-5-vPpipeπdin-1 -vO-2-hydroxypropan-1 -one: The rest of the synthesis is similar to the synthesis of (R)-1 -(4-(6-acetyl-7-amino-3-{6- phenylpyridin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5-yi)piperidin-1 -yl)-2-hydroxypropan-1 -one.
Synthesis of 1 -( 7-aminθ"5-(1 -methylpiperidin-4-vO-3-(6-phenylpyridin-3- yl)pyrazolori ,5-aipyrimidin-6-y[)ethanone
Figure imgf000438_0001
To a solution of 1-(7-amino-3-(6-phenylpyridin-3-yt)-5-(piperidin-4-yl)pyrazolo[1 ,5- a]pyrimidin-6-yl)ethanone (25 mg, 0.06 mmoi), DiEA (52 μl_, 0.3 mmol) in DMF (1 ml_) was added Mei (15 μL, 0.24 mmol). The reaction mixture was stirred for one hour and directly purified by HPLC to provide the title compound. LC/MS RT = 2.76 min. Mass calculated for M+H 427.2, observed 427.3.
General methods described above, compounds in the table 8-B can be synthesized.
TABLE- 8B
Figure imgf000438_0002
Figure imgf000439_0001
Figure imgf000440_0001
Figure imgf000441_0001
Figure imgf000442_0001
Figure imgf000443_0001
Figure imgf000444_0001
Figure imgf000445_0001
Figure imgf000446_0001
Figure imgf000447_0001
Figure imgf000448_0001
Figure imgf000449_0001
Figure imgf000450_0001
Figure imgf000451_0001
Figure imgf000452_0001
Figure imgf000453_0001
Figure imgf000454_0001
Figure imgf000455_0001
Figure imgf000456_0001
Figure imgf000457_0001
Figure imgf000458_0001
Figure imgf000459_0001
Figure imgf000460_0001
Figure imgf000461_0001
Figure imgf000462_0001
Figure imgf000463_0001
Figure imgf000464_0001
Figure imgf000465_0001
Figure imgf000466_0001
Figure imgf000467_0003
Figure imgf000467_0002
Synthesis of tert-butyl 4-(3-(6-(1 H-Dyrazol-1-yl)pyridin-3-yl)-7-(bisff2-(trimethylsilyl) ethoxy)methyl)amino)pyrazolof1 ,5-aiPyrimidin-5-yl)piDeridine-1-carboxylate:
Figure imgf000467_0001
To tert-butyl 4-(7-(bis((2-(iximethylsilyl)ethoxy)methyl)amino)-3-(6-chloropyridin-3- yl)pyrazolo[l,5-a]pyrimidin-5-yi)piperidine-1-carboxylate (2.25g, 3.3mmol) in Dioxane (24ml) and H2O (6mi) was added the 2-fluorophenyIboronic acid (0.92g, 6.6mmol), PdCl2(dppf)2.CH2Cl2 (0.32g, 0.39mmol) and K3PO4-H2O ( L7g, 8.2mmol). The reaction was heated at 100 °C for 16 hours, at which time LC/MS analysis confirmed full consumption of starting material. On cooling, H2O (100ml) and EtOAc (100ml) were added and organics were extracted with EtOAc (4 x 50ml), dried (Na2SO4) and concentrated in vacuo to give a crude product. Gradient column chromatography on silica eluting with 0% to 100% EtOAc/hexanes gave tert-butyl 4-(7-(bis((2-(trimethylsilyl) ethoxy)methyl)amino)-3-(6-(2- fluoropheny!)pyridin-3-yi)pyrazoio[i,5-a]pyrimidin-5-yl)piperidine-l-carboxylate (2.3g, 95%). By essentially the same procedure as to other related compound, different derivatives of above compound is prepared (Table-SB ).
Synthesis of tert-butyl 4-(3-(6-(1 H-pyrazol-1-yl)pyridin-3-yl)-7-(bisff2-(trimethylsilyl) ethoxyϊmethyltøminotøyrazoloH .5-aipyrimidin-5-vQpiperidine-i -carboxylate:
Figure imgf000468_0001
By essentially the same procedure as above, tert-butyl 4-(3-(6-(1H-pyrazol-1-yl)pyridin-3-yl)- 7-(bis((2-(trimethylsiiyl) ethoxy)methyl)amino) pyrazolo[ 1 ,5-a]pyrimidin-5-yl)piperidine-1- carboxylate is prepared.
Synthesis of (RH -(4-(7-aminQ-6-(2-hydroxyacetyl)-3-(6-phenylpyridin-3- yl)pyrazolo[1 ,5-alpyrimidin-5-yl)piperidin-1 -yl)-2-hydroxypropan-1 -one:
Figure imgf000468_0002
Step-1
Synthesis of 1 -(7-amino-3-(6-phenylpyridin-3-yl)-5-(piperidin-4-yl)pyrazoloπ .5- a1pyrimidin-6-yl)-2-hydroxyethanone: A solution of oxone (1.9g, 3.1mmol), trifluoroacetic anhydride (1.1ml, 7.9mmoi) and H2O (5.7ml) was stirred at 40°C for 7 h, and then cooled to room temperature. To this a solution of tert-butyl 4-(6-acetyl-7-(bis((2-(trimethylsiIyi)ethoxy)methyl)amino)-3-(6-phenylpyridin-3- yl)pyrazolo[t,5-a]pyrimidin-5-yl)piperidine-l-carboxylate (0.87g, 1.13mmol) and PhI (0.03ml, 0.23mmol) in acetonitrile (17ml) was added. The resulting solution was stirred at 90°C for 15 h. On cooling, the reaction mixture was filtered and solids were washed with DCM. It was then concentrated in vacuo to give a crude product which was submitted to the analytical group for purification to afford the desired l-(7-amino-3-(6-phenyIpyridin-3-yl)-5-(piperidin-4- yl)pyrazolo[ 1,5-a]pyrimidin-6-yl)-2-hydroxyethanone (94.2mg, 17%).
Step-2:
Synthesis of (RH -(4-(7-amino-6-(2-hydroxyacetyl)-3-(6-phenylpyridin-3- yl)pyrazoioM ,5-aipyrimidin-5-yl)piperidin-1 -yl)-2-hydroxypropan-1 -one:
A mixture of (R)-2-hydroxypropanoic acid (13.8mg, O.lβmraoi), EDCI (46mg, 0.24mmol), and 1-hydroxybenzotriazole (16.22mg, 0.12mmol) in DMF (2ml) was stirred at room temperature for lOmin. To this l-(7-amino-3-(6-phenylpyridin-3-yl)-5-(piperidin-4- y!)pyrazolo[1,5-a]pyrimidin-6-yl)-2-hydroxyethanone (58.77mg, 0.12mmol) was added followed by N,N-diisopropylethylamine (0. ImI, 0.6mmol). It was stirred further for 20min at room temperature at which time LC/MS analysis confirmed full consumption of starting material. This crude compound was submitted to the analytical group for purification to afford the desired (R)-1-(4-(7-amino-6-(2-hydroxyacetyl)-3-(6-phenylpyridin-3-yl)pyrazolo[ 1 ,5- a]pyrimidin-5-yl)piperidin-l-yl)-2-hydroxypropan-l -one. LCMS: 3.27 mins, m/z = 501.0 (MH+).
Synthesis of (R)-1-(7-amino-5-(1-(5-(1 -hydroxyethyl)-1 ,3.4-oxadiazol-2-yl)piperidin-4- yl)-3-(6-phenylpyridin-3-yl)pyrazoio[1 ,5-alpyrimidin-6-yl)ethaπone:
Figure imgf000470_0001
Step 1
Preparation of benzyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3- iodopyrazoioπ ^-alpyrimidin-5-yPpiperidine-i -carboxylate:
Figure imgf000470_0002
To a solution of benzyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1,5- a]pyrimidin-5-yl)piperidine-1-carboxylate (2.27g, 3.7 mmol) in acetonitriie (15 mL) was added NIS (833 mg, 1 eq}. After stirring 1.5 hour, solvent was removed and the residue was purified on silica gel. Elution with EtOAc/Hexanes (0-50%) gave the title compound (2.38 g, 86%) as gum.
Step 2
Preparation of benzyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)pyrazo[ori ,5-alpyrimidin-5-yl)piperidine-1 -carboxylate:
Figure imgf000470_0003
To a pressure tube were charged benzyl 4-(7-(bis((2-
(trimethylsilyl)ethoxyJmethyl)aminoJ-3-iodopyrazoloCI^-alpyrimidin-5-yl)piperidine-1- carboxylate (1 g, 2.85 mmol), 2-phenyl-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)pyridine (1.2g, 4.1 mmol), Pd(PPh^)4 (164 mg, 0.14 mmol), DME (20 mL) and 2 M IMa2CO3 (10 mL). The mixture was briefly degassed with Argon and the tube was capped and heated at 80 °C for 15 hours. On cooling, H2O (40 mL) and EtOAc (100 mL) and aqueous layer was extracted with EtOAc (3x) and combined organic layers were washed with brine once and dried (MgSO4). After concentration in vacuo the residue was purified on silica gel. Eiution with EtOAc/Hexanes (0-30%) gave the title compound (1.33 g, 61%).
Step 3
Preparation of 3-(6-phenylpyridin-3-yl)-5-(piperidin-4-yl)-NiN-bis(f2- ftrimethylsiiyl)ethoxy)methyl)pyrazolo[1 ,5-aipyrimidin-7-amine:
Figure imgf000471_0001
A mixture of benzyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-a3pyrimidin-5-yi)piperidine-1-carboxylate (1.33 g, 1.77 mmol), 10% Pd/C (170 mg) in EtOAc (10 mL) was stirred under hydrogen (balloon pressure) for two days, and then at 45 °C for 15 hours. After filtration and washing with EtOAc 4 times the title product (1 g, 93%) was obtained.
Step 4
Preparation of (R)-2-(benzyloxy)propanehydrazide;
Figure imgf000471_0002
A mixture of (R)-methyl 2-(benzyloxy)propanoate (1g, 5.15 mmol), hydrazine monohydrate (250 ul) in MeOH ( 14 mL) was refluxed 3 hours. Solvent was removed under reduced pressure and the residue was purified on silica gel. Eiution with EtOAc yielded title compound (640 mg, 64%).
Step S
Preparation of (RV5-M -(benzyloxy)ethyh-i ,3.4-oxadiazol-2(3/-fl-one:
Figure imgf000472_0001
To a solution of (R)-2-(benzyloxy)propanehydrazide (194 mg, 1 mmol) in anhydrous THF ( 2 mL) was added CDI (178 mg, 1.1 mmol) and the resulting mixture was allowed to stir one hour. . Solvent was removed under reduced pressure and the residue was purified on silica gel. Elution with EtOAc yielded title product.
Step 6
Preparation of (R)-5-(1-(5-(1-(benzyloxy)ethyl)-1 ,3.4-oxadiazol-2-yl)piperidin-4-yl)-3-
(6-phenylpyridin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5- alpyrimidin-7-amine:
Figure imgf000472_0002
To a stirred solution of (R)-5-(1-(benzyloxy)ethyl)-1 ,3,4-oxadiazol-2(3H)-one {44 mg, 0.2 mmol), 3-(6-phenyipyridin-3-yl)-5-(piperidin-4-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a3pyrimidin-7-amine (150 mg, 0.24 mmol) and DIEA {70 ul, 0.4 mmol) in DMF (1.4 mL) was added BOP (97 mg,0.22 mmol) and the resulting solution was allowed to stir overnight. Water was added and was extracted with EtOAc (3x) and combined organic layers were washed with water (3x), brine once and dried (MgSO4). After concentration in vacuo the residue was purified on silica gel. Elution with EtOAc/Hexanes (0-10%) gave the title compound (142 mg, 85 %).
Step 7
Preparation of I RH -(5-(4-( 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)pipehdin-1-yl)-1 ,3,4-oxadiazol-2- yl)ethanol:
Figure imgf000473_0002
2
A mixture of (R)-5-(1-(5-(1 -(benzyloxy)ethyl)-1 ,3,4-oxadiazoi-2-yl)piperidin-4-yl)-3-{6- phenylpyπdin-3-yl)-Λ/,Λ/-bis{(2-(trimethylstlyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrirτiidin-7- amine (105 mg), 20% Pd(OH)2/C (150 mg) in MeOH {4 ml_) was stirred under hydrogen (60 psi) overnight. After filtration, washing with EtOAc 4 times and concentration in vacuo the residue was purified on siiica gel. Elution with EtOAc/Hexanes (0-100%) gave the title compound (32 mg,43%).
Step 8
Preparation of (RH -(5-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)aminoV6-bromo-3-
(6-phenylpyridin-3-yl)pyrazolof 1 ,5-a1pyrimidin-5-vQpiperidin-1 -yl)-1 ,3,4-oxadiazol-2- yl)ethanol:
Figure imgf000473_0001
To a solution of (R)-1 -(5-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidin-1 -yl)-1 ,3,4-oxadiazol-2- yl)ethanol (32 mg, 0.04 mmo!) in acetonitriie (1 mi_) was added NBS (7 mg, 0.04 mmol) and the resulting solution was stirred for 10 minutes and concentrated, the residue was purified on silica gel. Elution with EtOAc provided a relatively pure product (9 mg).
Figure imgf000474_0001
Step 9
Preparation of (R)-1-(7-amino-5-(1-(5-(1-hydroxyethyl)-1 ,3,4-oxadiazol-2-yl)piperidin-
4-yl)-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-6-yl)ethanone:
Figure imgf000474_0002
To a pressure tube were charged (R)-1-(5-(4-(7-(bis({2- (trimethylstlyl)ethoxy)methyl)amino)--6-brorno-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5- a]pyhmidin-5-yl)piperidin-1-yI)-1 ,3,4-oxadiazol-2-yl)ethanol (9 mg,~0.01 mmol), Pd(PPh3)4 (10 mg, 0.087 mmoi), tributyl{-ethoxy-vinyl)tin (8 ui, 0.02 mmol) and dioxane (2 mL). The resulting mixture was degassed with Argon briefly, capped with a Teflon cap and stirred at 100 °C overnight. On cooling, the solvent was rotoevaporated, and the crude was redissolved in EtOAc (10 ml), washed with 0.5 M KF solution (1 x 2 ml), water (1 x 3 ml), brine (1 x 3 ml), and dried over MgSO4. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with EtOAc/Hexanes (0-100%) gave the intermediate eno! ether which was re-dissolve in dioxane (2 mL) and cooled to 0 °C and treated aqueous 4N HCI (0.2 mL). After the reaction mixture was warmed up to room temperature and stirred for 4 hours, solvents were removed and the residue was directly purified by HPLC to provide the title compound. Synthesis of (R)-7-amino-5-(1 -(2,3-dihydroxypropanoyl)piperidin-4-yl)-3-(6- phenylpyridin-3-yl)pyrazoio, ,5-alpyrimidine-6-carbonitrile:
Figure imgf000475_0001
Step 1
Preparation of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-cvano-3-(6- phenv[pyridin-3-yl)pyrazoiori ,5-alpyrimidin-5-yl)Diperidine-1 -carboxylate:
Figure imgf000475_0002
To a Schlenk tube were charged compound tert-butyl 4-(7-{bis((2- (trimethylsilyl)ethoxy)methy!)amino)-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)piperidine-1 -carboxylate (81 mg, 0.1 mmol), Bu3SnCN (47 mg, 1.5 eq.), Pd(PPh3J4 (23 mg, 0.2 eq.), Bis(tri-f-butylphosphine)palladium (0) (10 mg, 0.2 eq.). The tube was evacuated and charged with Ar for three cycles. Dioxane (3 ml) was added; the tube was capped and heated at 160 C with stirring for one hour. After cooling, the mixture was diluted with EtOAc and washed with brine once. Organic layer separated, dried over MgSO4 and concentrated. The residue was purified on silica gel. Elution with EtOAc/hexane (0-25%) gave the desired title product (63 mg, 84%).
Step 2
Preparation of 7-amino-3-(6-phenylpyridin-3-yl)-5-(piperidin-4-yl)pyrazolof 1 ,5- aipyrimidine-6-carbonitrile:
Figure imgf000476_0001
Tert-butyl 4-{7-(bis((2-(trtmethylsilyl)ethoxy)methyl)amino)-6-cyano-3-(6-pheny!pyridin- 3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperidine-1 -carboxylate (190 mg) was treated with TFA/water(95:5, 3 ml) with stirring for 10 minutes. The reaction mixture was concentrated and lyophilized to yield the title compound as TFA salt.
Step 3
Preparation of (R)-7-amino-5-(1 -(2,3-dihydroxypropanoyl)piperidin-4-vO-3-(6- phenylpyridin-3-yl)pyrazoloF1 ,5-alpyrimidine-6-carbonitrile:
DMF
Figure imgf000476_0002
To a solution of 7-amino-3-(6-phenyJpyridin-3-yl)-5-(piperidin-4-yl)pyrazolo[1 ,5- a]pyrimidine-6-carbonitrile TFA salt (0.1 mmol), DIEA (87μL, 0.5 mmol) in DMF (2 ml_) was added a solution of (R)-2,3-dihydroxypropanoic acid (11 mg, 0.1 mmol), HOBt (14 mg, 0.1 mmoi), EDCl (29 mg, 0.15 mmol) in DMF (1 mL). The reaction was stirred at rt for 2.5 hour and the mixture was directly purified by HPLC to furnish the title compound. LC/MS RT = 2.86 min. Mass calculated for M+H 484.2, observed 484.3.
C
Figure imgf000477_0001
Scheme 27
Synthesis of pyrazoloH ,5-aipyrimidine-5,7-diol:
Diethyl malonate NaOEt, EtOH
Figure imgf000477_0002
To 1 H-pyrazol-3-amine (12.3 g, 148.0 mmol) in EtOH (50 mL) was added diethy! malonate (25.0 mL, 164.7 mmol), 21wt% NaOEt in EtOH (1 10 mL, 294.6 mmol) and additional EtOH (50 mL). The resulting reaction mixture was then heated at 80 °C under an atmosphere of argon for 16 hours, at which time the reaction was allowed to cool to room temperature. The reaction mixture was then concentrated in vacuo until almost dry, before H2O (500 mL) was added. Vigorous stirring aided the dissolution of solids, at which time cone. HCI was added until pH~2 was attained (precipitate formed). The precipitate was collected and dried by vacuum fiitration giving pyrazolo[1 ,5-a]pyrimidine-5,7-diol as a tan solid (17.13 g, 113.4 mmol, 77%).
Synthesis of 5.7-dichloropyrazolo[1 ,5-aipyrimidine
Figure imgf000477_0003
To pyrazolo[1.5-alpyrimidine-5J-diol (9.6 g, 63.5 mmol) in a 500 mL flask was added POCI3 (125 mL, 1341 .1 mmol). The flask was then cooled to 0 °C and N, N- dimethylaniline (22 mL, 173.6 mmol) was carefully added. On warming to room temperature, the reaction mixture was then heated at 60 °C under an atmosphere of argon for 16 hours. On cooling, the reaction mixture was concentrated in vacuo to give a brown viscous liquid. This brown viscous liquid was carefully poured onto ice and allowed to warm to room temperature overnight. To the brown solution was carefully added saturated NaHCO3 solution until no further effervescence was observed and pH ~ 8 was attained. Organics were then extracted with CH2CI2 (4 x 50 mL), dried (Na2SO4) and concentrated in vacυo to give a brown liquid (29.8 g). Gradient column chromatography on silica eluting with 50% CH2C!2/hexanes (to elute aniline) followed by 75% CH2CI2/hexanes (to elute product) gave 5,7-dichloropyrazoio[1 ,5-a]pyrimidine as a white solid (7.7 g, 40.8 mmol, 64%).
Synthesis of 5-chloropyrazoloH ,5-a]pyrimidin-7-amine
Figure imgf000478_0001
To 5,7-dichloropyrazolo[1 ,5-a]pyrimidine (7.6 g, 40.4 mmoi) in a sealed vessel was added NH4OH (100 mL). The vessel was then sealed and heated at 85 °C for 2.5 .. hours, at which time the consistency of the white solid had changed (from foamy white solid to free-flowing white solid). The vessel was removed from the heat source and allowed to cool to room temperature overnight. On cooling, the contents of the vessel were collected and dried by vacuum filtration giving 5-chloropyrazolo[1 ,5-a]pyrimidin- 7-amine as a yellow-tinged white solid (6.8 g, 40.3 mmol, 100%).
Synthesis of 5-chloro-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolof 1 ,5-aipyrimidin- 7-amine
Figure imgf000478_0002
To 5-chloropyrazoio[1 ,5-a]pyrimidin-7-amine (6.7 g, 39.7 mmol) in CH2CI2 (30 ml_) was added N,N-diisopropylethylamine (48.0 mL, 275.6 mmol) followed by 2- (Trimethylsilyl)ethoxymethyl chloride (25.0 mL, 141.7 mmol). The reaction mixture was heated at 45 °C for 3 hours before being allowed to cool to room temperature. The reaction mixture was then poured into a separatory funnel containing ~100 mL saturated NaHCO3 solution and CH2CI2 (50 mL). Organics were then extracted with CH2CI2 (4 x 50 mL), dried (Na2SO4) and concentrated in vacuo to give a thick orange liquid (33.8 g). Gradient column chromatography on silica eluting with 5% to 15% EtOAc/hexanes gave crude 5-chloro-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine as a colorless liquid (18.7 Q).
Synthesis of 5-chloro-3-iodo-N,N-bis((2-(trimethylsi[yπethoxy)methyl)pyrazolori .5- aipyrimidin-7-amine
Figure imgf000479_0001
To crude 5-chloro-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazoIo[1 ,5-a]pyrimidin-7- amine (7.9 g) in CH3CN (100 mL) was added N-iodosuccinimide (4.3 g, 19.2 mmol) and the resulting mixture was stirred at room temperature for 30 mins, at which time LC/MS confirmed full conversion of starting material to product. Saturated sodium thiosulfate solution (-20 mL) was added and stirring continued for 5 minutes before the mixture was transferred to a separatory funnel using CH2Cb (30 mL) and H2O (30 mL). Brine (50 mL) was added and organics were extracted with CH2CI2 (4 x 40 mL), dried (Na2SO4) and concentrated in vacuo to give crude 5-chloro-3-iodo-N,N-bis({2- (trimethylsilyl)ethoxy)methyl)pyrazoio[1,5-a]pyrimidin-7-amine as a Sight brown liquid (10.4 g). LCMS: 2.95 mins, m/z = 555.1 (MH+).
Synthesis of 5-chloro-3-(quinolin-3-vO-N,N-bisf(2- ftrimethylsilyl)ethoxy)methyl)pyrazolon .5-alpyrimidin-7-amine
Figure imgf000480_0001
To crude 5-chloro-3-iodo-N,N-bis((2-(tnmethytsilyI)ethoxy)methyl)pyrazolo[1 ,5- a]pyrimi,din-7-amine (7.1 g) in DME (120 mL) and H2O (15 ml_) was added the quinoline boronic acid (2.4 g, 14.1 mmol), PdCI2(dppf)2 (1.0 g, 1.2 mmol) and K3PO4-H2O (5.4 g, 25.6 mmol). The reaction mixture was heated at 60 °C for 2 hours, at which time LC/MS analysis confirmed full consumption of starting material. On cooling, H2O (40 mL) and EtOAc (100 mL) were added and organics were extracted with EtOAc (4 x 50 mL), dried (Na2SO4) and concentrated in vacuo to give a brown oil. Gradient column chromatography on silica eluting with 10% to 60% EtOAc/hexanes gave 5-chloro-3-(quinolin-3-yl)-N,N-bis((2-(thmethylsiiyl)ethoxy)methyl)pyrazoio[1 J5- a]pyrimidin-7-amine as a light yellow solid (4.1 g, 7.4 mmol, 65% over three steps). LCMS: 2.62 mins, m/z = 556.2 (MH+).
Figure imgf000480_0002
SCHEME-28
Synthesis of 5-morpholino-3-(αuinolin-3-yl)pyrazoloM .5-aipyrimidin-7-amine
Figure imgf000480_0003
To a 2-5 mL microwave vessel is charged 5~chloro~3-(quinolin-3-yl)~N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine (72 μmol, 40 mg), N1N- diisopropylethylamine (220 μmol, 38 μl_), NMP (2 mL), and morpholine (220 μmol, 19 μL). The reaction vessel is flushed with agron and sealed. The reaction mixture was heated to 200° C in microwave synthesizer for 30 minutes. Upon completion, the NMP was removed in vacuo as an azeotropic using chlorobenzene. The crude product was purified via reverse-phase preparatory HPLC to yield 5-morpholino-3- (quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine as brown-yellow solid. (m+H = 347.31 , retention time = 2.97 min).
10
By essentially the same procedure given in the synthesis of 5-morpho!ino-3-(quinolin- 3-yl) pyrazolo[1 ,5-a]pyrimidin-7-amine, Schemes 27 & 28, the compounds listed in Table 9 can be prepared.
\5 Table 9
Figure imgf000481_0001
Figure imgf000482_0001
Figure imgf000483_0001
Synthesis of 1-(7-amino-3-(quinolin-3-yl)pyrazolo[1 ,5-aipyrimidin-5-yl)-1,4-ciiazepan-5- one
Figure imgf000484_0001
A mixture of 5-chIoro-3-(quinolin-3-yl)-N]N-bis((2-(trimethylsilyl)ethoxy) methyl)- pyrazolo-[1 ,5-a]pyrimidin-7-amine (50 mg, 0.09 mmoL), 1 ,4-diazepan-5-one (36 mg, 0.32 mmoL), EtN(SPr)2 (110 uL, 0.63 mmoL) in NMP (0.8 ml_) was heated at 185°C under microwave condition for 45 min. Purification by prep-LC afforded titled compound (32.3 mg, 85%) as its formic acid salt. LCMS tR = 2.49 Min. Mass calculated for, M+ 373.1 , observed LC/MS m/z 374.1 (M+H).
Synthesis of 1-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo|"1 ,5-a1pyrimidin-5-yl)-1 A- diazepan-5-one
Figure imgf000484_0002
A solution of NBS (14.6 mg, 0.082 mmoL) in CH3CN (0.5 mL) was added to a mixture of 1 -(7-amino-3-(quinolin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5-yi)-1 ,4-diazepan-5-one formic acid salt (32.3 mg, 0.077 mmoL) in CH3CN (3 mL). After stirring at room temperature for 10 min, the solvent was evaporated and the residue was purified by prep-LC to afford titled compound (22.6 mg, 59%). LCMS tR =2.78 Min. Mass calculated for, M+ 451.0, observed LC/MS m/z 452.0 (M+H).
Methyi 1 -(7-amino-3-(αuinolin-3-yl)pyrazoloH ,5-aipyrimidin-5-yl)pyrrolidine-3- carboxyiate
Figure imgf000485_0001
A mixture of 5-chloro-3-(quinolin-3-yl)-N,N-bis((2-(trimethylsiiyl)ethoxy) methyl)- pyrazolo-[1 ,5-a]pyrimidin-7-amine (55 mg, 0.10 mmoL), methyl pyrrolidine-3- carboxylate (52 mg, 0.30 mmoL) and EtN(IPr)2 (86 uL, 0.50 mmoL) in DMF (1.0 ml_) was heated at 18O°C under microwave condition for 45 min. The mixture was purified by prep-LC to afford the title compound. LC/MS RT = 3.30 min. Mass calculated for, M+H 389.17, observed 389.17.
1 -(7-Amino-3-(quinolin-3-yl)pyrazo!of 1 ,5-alpyrimidin-5-yl)pyrrolidine-3-carboxylic acid
Figure imgf000485_0002
To a solution of methyl 1-(7-amino-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)pyrrolidine-3-carboxylate (19 mg, 0.050 mmol) in a 2:1 mixture of THF and H2O (3 ml_) was added 1 N LiOH solution (0.25 ml_). The reaction mixture was stirred at rt overnight and concentrated in vacuo. The residue was purified by prep-LC to afford the title compound. LC/MS RT = 2.80 min. Mass calculated for, M+H 375.16, observed 375.15.
.1-(7-Amino-6-bromo-3-(Quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)pyrroiidine-3- carboxylic acid
Figure imgf000486_0001
To a solution of 1 -(7-amino-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)pyrroiidine-3- carboxylic acid (17 mg, 0.033 mmol) in a 2:1 mixture of CH3CN/MeOH (1.0 mL) was added NBS (6.2 mg, 0.033 mmol). The mixture was stirred at rt for 1 h then concentrated in vacuo. The residue was purified by prep-LC to afford the title compound. LC/MS RT = 3.27 min. Mass calculated for, M+H 453.07, observed 453.07
Synthesis of tetrahydro-imidazoH ,5-a]pyrazine-1 ,3-dione
Figure imgf000486_0002
Stepi :
Potassium cyanate (10 mmol, 811 mg) was added to 1 -benzyi-3-methyl piperazine-
1 ,3-dicarboxy!ate ( 2 mmol, 556 mg) dissolved in methanol-water, and the reaction mixture heated at 75 °C for 2hrs until LCMS showed the completion of the reaction.
The solvent was evaporated in vacuo and the crude product was purified by silica get column chromatography to yield the product, benzyl 1 ,3- dioxohexahydroimidazo[1 ,5- a]pyrazine-7(1 H)-carboxylate, ( 290mg, 50%). LCMS tR =2.50 min. Mass calculated for, M+ 289.11, observed LC/MS m/z 288.20 (M-H negative ion mode).
Step2;
To benzyl 1 , 3- dioxohexahydroimidazo[1 ,5-a]pyrazine-7(1 H)-carboxylate ( 1.0 mmol,
290 mg) in methanol was added 50 mg of Pd/C(10%). The solution was kept under hydrogen atmospheric by a balloon for 14 hrs at which time LCMS showed the completion of the reaction. The solution was filtered through celite and the solvent was evaporated in vacuo Xo dryness to yield, tetrahydroimidazo[1 ,5-a]pyrazine-1 ,3(2H,5H)- dione. LCMS tR =1.00 min. Mass calculated for, M+ 155.07, observed LC/MS m/z 156 (M+H).
By essentially the same procedure given in Scheme 28, the compounds listed in TablelO can be prepared. Table lO
Figure imgf000487_0001
Figure imgf000488_0001
Figure imgf000489_0001
Figure imgf000490_0001
Figure imgf000491_0001
Figure imgf000492_0001
Figure imgf000493_0001
Figure imgf000494_0001
Figure imgf000495_0001
Figure imgf000496_0001
Figure imgf000497_0001
Figure imgf000498_0001
Figure imgf000499_0001
Figure imgf000500_0001
6-bromo-5-morphoiino-3-(quinolin-3-yl)-N,N-bisf(2-(trimethyisilyl) ethoxy)methyl)pyrazolo[1 ,5-aiPyrimidin-7-amine
Figure imgf000501_0001
A solution of NBS (20.6 mg, 0.116 mmoL) in CH3CN (1 mL) was added to a mixture of 5-morphoiino-3-(quinolin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyra--olo[1 ,5- a]pyrimidin-7-amine (70.0 mg, 0.1 16 mmoL) in CH3CN (6 mL). After stirring at room temperature for 10 min, the reaction mixture was concentrated and purified by column chromatography to afford the desired compound. LCMS tR =2.91 Min (5 min run, UV 254nm). Mass calculated for, M+ 684.2, observed LC/MS m/z 685.2 (M+H).
Synthesis of 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-5-morphoiino-3-fquinolin-3- yl)pyrazolori .5-aipyrimidine-6-carbonitrile
Figure imgf000501_0002
A degassed mixture of 6-bromo-5-morpholino-3-(quinolin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine (35 mg, 0.051 mmoL), Bu3SnCN (32.3 mg, 0.10 mmoL}, Pd[P(t-Bu)3]2 (5.2 mg, 0.010 mmoL) in Dioxane (3 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and the solvent was evaporated in vacuo. Purification by column chromatography afforded the titled compound. LCMS tR = 2.76 Min (5 min run, UV 254nm)- Mass calculated for, M+ 631.3, observed LC/MS m/z 632.2 (M+H).
Synthesis of 7-amino-5-morpholino-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidine-6- carbonitrile
Figure imgf000502_0001
A mixture of 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-5-morpholino-3-(quinolin-3- yi)pyrazolo[1 ,5-a]pyrimtdine-6-carbonitπle (5 mg), EtOH (1 mL) and 3N HCi (1 mL) was heated at 6O°C until LCMS indicated the completed reaction. The mixture was cooled to room temperature and the solvent was evaporated in vacuo. Purification by prep-LC afforded the desired compound. LCMS tR = 3.12 Min (UV 254nm)- Mass calculated for, M+ 371.1 , observed LC/MS m/z 372.2 (M+H).
Synthesis of 5-morpholino-3-(quinolin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-aipyrimidin-7-amine
Figure imgf000502_0002
A mixture of 5-chloro-3-(quinolin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoio[l,5-a]pyrimidin-7-amine (77.9 mg, 0.14 mmoL), morpholine (36.7 mg, 0.42 mmoL), NaHCO3 (53 mg, 0.63 mmoL) in NMP (3 mL) was heated at 13O°C overnight. The mixture was cooled to room temperature and diluted with H2O and then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation of solvent afforded the crude displacement compound. Purification by column chromatography afforded 5-morpholino-3-(quinolin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methy[)pyrazolo[l,5-a]pyrimidin-7-amine (70 mg, 82%). LCMS tR = 2.72 Min (5 min run, UV 2sW- Mass calculated for, M+ 606.3, observed LC/MS m/z 607.2 (M+H).
Synthesis of 5-thiomorpholinodioxo-3-(quinolin-3-yl)-N, N-bis((2- (trimethylsilyl)ethoxy)methyJ)pyrazolof1 ,5-a1pyrimidin-7-amine
Figure imgf000503_0001
A mixture of 5-chloro-3-(quinolin-3-yl)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine (503.0 mg, 0.91 mmoL), thiomorpholine 1, 1-dioxide (367.1 mg, 2.72 mmoL), NaHCO3 (533.0 mg, 6.34 mmoL) in NMP (8 mL) was heated at 13O°C overnight. The mixture was cooled to room temperature and diluted with H2O and then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation of solvent afforded the crude displacement compound. Purification by column chromatography afforded compound 5j; thiomorpholinodioxo-3-(quinolin-3-yl)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine LCMS tR = 2.52 Min (5 min run, UV 254nm)< Mass calculated for, M+ 654.2, observed m/z 655.2 (M+H).
Synthesis of 5-morpholino-3-(quinolin-3-yl)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazo[oπ,5-aipyrimidin — 6-bromo-7-amine
Figure imgf000503_0002
A solution of NBS (20.6 mg, 0.116 mmoL) in CH3CN (1 mL) was added to a mixture of 5- morpholino-3-(quinolin-3-yl)-N,N-bis((2-(trimethy lsilyl)ethoxy)methyl)pyrazolo[ 1 ,5- a]pyrimidin-7-amine (70.0 mg, 0.116 mmoL) in CH3CN (6 mL). After stirring at room temperature for 10 min, the reaction mixture was concentrated and purified by column chromatography to afford 6-bromo-5-morphoIino-3-(quinolin-3-yl)-N,N-bis((2- (trimethyisiIyl)ethoxy)methyl)pyrazoIo[1,5-a]pyrimidin-7-amine. LCMS tR =2.91 Min (5 min run, UV 254nm). Mass calculated for, M+ 684.2, observed LC/MS m/z 685.2 (M+H). Synthesis of 5-thiomorpholinodioxo-3-(αuinolin-3-yl)-N,N-bis((2- (trimethylsilv[)ethoxy)methyl)pyrazoio|"1 ,5-aipyrimidin-6-bromo-7-amine
Figure imgf000504_0001
By essentially the same procedure given in Preparative Example 3- 1, 6-bromo-3-(quinolin-3- yl)-5-thiomorpholino-r, r-dioxide-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[I ,5- a]pyrimidin-7-amine can be prepared. LCMS IR = 2.90 Min (S min run, UV 2S4nm)- Mass calculated for, M+ 732.2, observed LC/MS m/z 733.2 (M+H).
Synthesis of 6-bromo-3-(6-fluoroquino[in-3-yl)-5-thiomorpholin-dioxo-pyrazoiori ,5- aipyrimidin-7-amine:
Figure imgf000504_0002
SCHEME-29
Part A:
Synthesis of 3-iodo-5-thiomorpholin-4-dioxo-N,N-bisf(2-
(trimethylsilyl)ethoxy)methyQpyrazolori ,5-a]pyrimidin-7-amine: A mixture of compound 5-chloro-3-iodo-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (10.8 mmol, 6.00 g, preparation reported previously), thiomorpholine 1,1-dioxide (32.4 mmol, 4.38 g), and NaHCO3 (48.6 mmol, 4.08 g) in NMP (50 mL) was heated at 130 °C for 28 h. LCMS showed nearly complete consumption of A. The reaction mixture was diluted with EtOAc, washed with H2O and brine, dried over Na2SO4, and concentrated. The crude product was purified by a SiO2 column (0-30% EtOAc/Hexanes) to afford compound 3-iodo-5-thiomorpholin-4-dioxo- N,N-bis((2-(trimethyisiiyI)ethoxy)methy[)pyrazoIo[1,5-a]pyrimidin-7-amine as a pale yellow solid (6.15 g, 87%). HPLC-MS TR = 2.70, min (UV 254 nm, 5 min method); mass calculated for formula C22H40IN5O4SSi2653.1 , observed LCMS m/z 654.1 (M+H).
Part B:
Synthesis of 3-(6-fluoroαuinolin-3-yl)-5-thiomorpholin-4'-dioxo-N,N-bts((2-
(trimethylsilyl)ethoxy)methyi)pyrazolori ,5-a1pyrimidin-7-amine: A mixture of compound 3-iodo-5-thiomorpholin-4-dioxo-N,N-bis((2-
(trimethylsi!yl)ethoxy)methyl)pyrazoio[1,5-a]pyrimidin-7-amine (0.551 mmol, 360 mg), boronate 7 (0.827 mmol, prepared in situ, see below), PdCl2CdPPf)CH2Cl2 (0.055 mmol, 45.0 mg), and K2CO3 (1.65 mmoi, 229 mg) in DMEyH2O (5/1 mL) was degassed and then heated at 90 °C for 24 h. The reaction mixture was diluted with EtOAc and filtered. The filtrate was concentrated and purified by a SiO2 column (0-50% EtOAc/Hexanes) to afford compound 3- (6-fluoroquinolin-3-yl)-5-thiomorphoIin-4'-dioxo-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine as a yellow solid (260 mg, 70%). HPLC-MS TR = 2.39 min (UV 254 nm, 5 min method); mass calculated for formula C3 IH45FN6O4SSi2672.3, observed LCMS m/z 673.2 (M+H).
Synthesis of 6-fluoro-3-(4,4,5,5-tetramethyl-1 ,3.2-dioxaborolan-2-yl)Quinoliπe:
Figure imgf000505_0001
A mixture of the corresponding 3-bromo-6-fluoro quinoline (0.827 mmol, 187 mg, 1.0 eq), bis(pinacolato)diboron ( 12 eq) ,PdCl2(dppf)CH2Cl2 (0.1 eq), and KOAc (3.0 eq) in dioxane (5 mL) was degassed and then heated at 80 °C for 16 h. All the volatiles were removed under reduced pressure and the crude product 6-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)quinoline was used without further purification.
Part C:
3-(6-fluoroαuinolin-3-yl)-5-thiomorpholin-4'-dioxo- pyrazolo[1.5-a1pyrimidin-7-amine :
To a solution of compound 3-(6-fluoroquino[in-3-yl)-5-thiomorphoIin-4'-dioxo-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (0.169 mmol, 156 mg) in EtOH (4 mL) was added 4 N HCl in dioxane ( 1 mL) and the resulting mixture was heated at 65 °C for 1 h. All the volatiles were removed under reduced pressure. The residue, which has a very poor solubility in DMSO, was diluted with a small amount of DMSO and basicified with NaHCO3. The resulting slurry was stirred vigorously overnight and filtered. The precipitates were washed with H2O and MeOH to afford compound 3-(6-fluoroquinolin-3-yl)-5- thiomorphoiin-4'-dioxo- pyrazolo[l ,5-a]pyrimidin-7-amine as a pale yellow solid (60.1 mg, 86%). HPLC-MS TR = 1.17 min (UV 254 nm, 5 min method); mass calculated for formula CI9HI7FN6O2S 412.1, observed LCMS m/z 413.0 (M+H).
Part D: 3-(6-fluoroquinolin-3-yl)-5-thiornorpholin-4'-dioxo-6-brorno-pyrazolori .5-aipyrimidin-7- amine :
To a slurry of compound 3-(6-fluoroquinolin-3-yl)-5-thiomorphoUn-4'-dioxo- pyrazolo[l,5- a]pyrimidin-7-amine (0.0659 mmol, 27.2 mg) in CH3CN was added NBS (1.0 eq, in a stock solution in CH3CN) and the resulting mixture was stirred at rt for ! h. The reaction solution was concentrated and purified by prep-HPLC to afford compound 3-(6-fluoroquinolin-3-yl)-5- thiomorpholin-4'-dioxo-6-bromo-pyrazolo[1,5-a]pyrimidin-7-amine as a pale yellow solid (6.1 mg, 19%). HPLC-MS TR = 1.56 min (UV 254 nm, 5 min method); mass calculated for formula C19Hi6BrFN6O2S 490.0, observed LCMS m/z 491.0 (M+H).
By essentially the same procedures given in Preparative Example (scheme-29)Part B, Part C and Part D, compounds 10.65-10.85 given in Table 1OA can be prepared. TABLE ■ IQA
Figure imgf000507_0001
Figure imgf000508_0001
Figure imgf000509_0001
Figure imgf000510_0002
Synthesis of 3-(2,3'-bipyridin-5-yl)-5-thiomorphoiino-4-dioxo-N, N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-alpyrimidin-7-amine:
Figure imgf000510_0001
SCHEME-30
Part A:
3-(6-chloropyridin-3-yl)-5-thionnorpholino4-dioxo-N,N-bis((2- (trimethylsilyi)ethoxy)methyl)pyrazolo[1,5-aipyrimidin-7-amine:
Title compound was prepared from compound 3-iodo-5-εhiomorpholino4'-dioxo-N,N-bis((2- (trimethy I si Iy i )ethox y ) methy l)py razolo [ 1 ,5 -a] py rimidin-7-amine and 6-chloro -3 - pyridineboronic acid pinacol ester using the coupling conditions described in Part B of Example 1. EPLC-MS TR = 2.89 min (UV 254 ran, 5 min method); mass calculated for formula C27H43ClN6O4SSi2 638.2, observed LCMS m/z 639.1 (M+H).
Part B: 3-(2.3'-bipyridin-5-yl)-5-thiomorphoiino-4-dioxo-N.N-bisf(2- (trimethylsilyl)ethoxy)methyi)pyrazolori ,5-a1pyrinnidin"7-amine: A mixture of 3-(6-chloropyridin-3-yl)-5-thiomorpholino4-dioxo-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine (0.258 mmol, 165 mg), pyridine-3-ylboronic acid (0.516 mmol, 63.4 mg), PdCI2(dppf)CH2Cl2 (0.026 mmol, 21.2 mg), and K2CO3 (0.774 mmol, 107 mg) in OMEfH2O (2/0.4 mL) was degassed and then heated at 150 °C under microwave radiation for 1 h. The reaction mixture was diluted with EtOAc and filtered. The filtrate was concentrated and purified by a SiO2 column (20-100% EtOAc/Hexanes) to afford compound 3-(2,3'-bipyridin-5-yl)-5-thiomorpholino-4-dioxo-N,N- bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine as a pale yellow solid (161 mg, 91%). HPLC-MS TR = 2.15 min (UV 254 nm, 5 min method); mass calculated for formula C32H47N7O4SSi2681.3, observed LCMS m/z 682.3 (M+H).
By essentially the same procedures given in Preparative above Example(scheme-30) Compounds given in Table 1OB can be prepared.
Table 1OB
Figure imgf000511_0001
Figure imgf000512_0002
Synthesis of 3-(3,3'-bipyriclin-5-yl)-5-thiomorpholin-4'-dioxo-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-alpyrimidin-7-amine;
Figure imgf000512_0001
SCHEME-31
Part A:
3-(5-bromopyridin-3-yl)-5-thiomorpholin-4'-dSoxo-N,N-bis((2- ftrimethylsilyl)ethoxy)methyl)pyrazolof1 ,5-aipyrimidSn-7-amine: Compound 3-(5-bromopyridin-3-yI)-5-thiomorpholin-4'-dioxo-N,N-bis((2- (trimethylsϋyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine was prepared from compound 3-iodo-5-thiomorphoUno4'-dioxo-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5- a]pyrimidin-7-amine and 5-bromo-3-pyridineboronic acid pinacol ester using the coupling conditions described in Part B of Example 1. HPLC-MS TR = 2.62 min (UV 254 nm, 5 min method); mass calculated for formula C2TH43BrN6O4SSi2 682.2, observed LCMS m/z 683.0 (M+H).
Part B: 3-(3,3'-bipyridin-5-yl)-5-thiomorDholin-4'-dioxo-N.N-bis((2-
(trimethylsilvOethoxy)nnethyl)pyrazoloπ,5-alpyrimidin-7-amine:
Compound, 3-(3,3'-bipyridin-5-yl)-5-thiomorpholin-4'-dioxo-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine was prepared from 3-(5- bromopyridin-3-yi)-5-thiomorpholin-4'-dioxo-N,N-bis((2- trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine using the coupling conditions described in Part B of Preparative Example 3. HPLC-MS TR = 2.50 min (UV 254 nm, 5 min method); mass calculated for formula C27H43BrN6O4SSi2 681.3, observed LCMS m/z 682.2 (M+H). By essentially the same procedures given in Preparative Example above( Scheme-31 ), compounds given in Table- 1OC can be prepared
Table IQC
Figure imgf000513_0001
1-(7-amino-5-morpholino-3-(quinolin-3-yl)pyrazolori ,5-aipyrimiciin-6-yl)ethanone:
Figure imgf000514_0001
SCHEME-32
A degassed mixture of 6-bromo-5-morpholino-3-(quinolin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)niethyl)pyrazolo[l,5-a]pyrimidin-7-amine (141 mg, 0.20mmoL), tributyl(l-ethoxyvinyl)tin (112 mg, 0.31 mmoL), Pd(PPh3)4 (24 mg, 0.02 mmoL) in Dioxane (4 mL) was heated at 100°C for 2 days. The mixture was cooled to room temperature and filtered through short plug of 10% KF/Siθ2. The crude Stille coupling product obtained after concentrating the filtrate was then treated with 3mL of 1: 1 mixture of TFA and H2O at room temperature for 30 min, Concentration and purification by prep-LC afforded l-(7-amino-5- morpholino-3-(quinolin-3-yl)pyrazolo[1,5-a]pyrimidin-6-yl)ethanone. LCMS tR = 3.32 Min. Mass calculated for, M+ 388.1, observed LC/MS mJz 389.1 (M+H).
By essentially the same procedure given in Preparative above(Scheme-32), compounds in the table- 10-D can be synthesized.
TABLE-10D
Figure imgf000514_0002
Figure imgf000515_0001
Figure imgf000516_0002
Synthesis of 1-(7-amino-3-(1-phenyl-1 H-pyrazoi-4-yl)-5-thiomorpholin-4'-dioxo- pyrazoion ,5-aipyrimidin-6--yl)etharione:
Figure imgf000516_0001
SCHEME- 33
Part A: 10 3-(1 H-pyrazol-4-ylV5-thiomorpholin-4'-dioxo-N.N-bisf(2- ftrimethvlsilvπethoxvlmethvπpvrazoloFl5-aipvrimidin-y-amine:
A mixture of compound, 3-(5-bromopyridin-3-yl)-5-thiomorpholin-4'-dioxo-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine (1.90 mmol, 1.24 g) 1-Boc- ] 5 pyrazole-4-boronic acid pinacol ester (2.85 mmol, 838 mg), Pd(PPh3)4 (0.19 mmol, 220 mg), and K2CO3 (5.70 mmol, 788 mg) in dioxane/H2O (20/5 mL) was degassed and heated at 80 °C overnight using an oil bath. Then the reaction mixture was heated at 150 °C under microwave radiation for 1 h. After cooling to rt, the reaction mixture was diluted with EtOAc, washed with H2O and brine, dried over Na2SO4, and concentrated. The residue was purified by a SiO2 column (20-100% EtOAc/Hexanes, Rf = 0.5 in 100% EtOAc/Hexanes) to afford compound, 3- (1H-pyrazol-4-yl)-5-thiomorpholin-4'-dioxo-N,N-bis((2- (trimethylsilyl)ethoxy)methy!)pyrazolo[1,5-a]pyrirnidin-7-amine as a brown solid (358 rag, 32%). HPLC-MS TR = 2.39 min (UV 254 nm, 5 min method); mass calculated for formula C25H43N7O4SSi2593.3, observed LCMS m/z 594.2 (M+H).
Part B: 3-(1 -pheπyl-1 H-pyrazol-4-yl)-5-thiomorprιoiin-4'-dioxo-N,N-bis(f2- (trimethyisilvnethoxy)methyl)pyrazolori ,5-a1pyrimiciin-7-aiinine: A mixture compound, 3-(1H-pyrazol-4-yl)-5-thiomorpholin-4'-dioxo-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[l ,5-a]pyrimidin-7-amine (0.603 mmol, 358 mg), iodobenzene (0.904 mmol, 101 μL), CuI (0.0603 mmol, 11.5 mg), N,N'-dimethyl- ethylenediamine (0.241 mmol, 25.9 μL) and K3PO4 (1.27 mmol, 269 mg) in toluene (6 mL) was stirred at 110 °C under Ar2 for 24 h. After cooling to rt, the crude mixture was diluted with EtOAc and filtered. The filtrate was concentrated and purified by a SiO2 column (0-50% EtOAc/Hexanes, Rf = 0.7 in 50% EtOAc/Hexanes) to afford the compound, 3-(l-phenyl-1H- pyrazol-4-yl)-5-thiomorpho!in-4'-dioxo-Ν,Ν-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine, as a pale yellow solid (365 mg, 90%). HPLC-MS TR = 2.77 min (UV 254 nm, 5 min method); mass calculated for formula C31H47N7O4SSi2 669.3, observed LCMS m/z 670.3 (M+H).
Part C: 6-bromo-3-(1 -phenyl-1 H-pyrazol-4-yl)-5-thiomorpholin-4'-dioxo-N.N-bis((2- (trirπethylsilyl)ethoxy)methyhpyrazolof1,5-aipyrimidin-7-amine:
To a solution of compound 3-(l-phenyl-1H-pyrazol-4-yl)-5-thiomorpholin-4'-dioxo-N,N- bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (0.545 mmol, 365 mg) in CH3CN/DCM (3/3 mL) was added NBS (0.545 mmol, 97 mg) and stirred at rt for 1 h. All the volatiles were removed under reduced pressure and the residue was purified by a SiO2 column (0-50% EtOAc/Hexanes, Rf = 0.8 in 50% EtOAc) to afford compound, 6-Bromo-3-(l- phenyl- 1H-pyrazoI-4-yl)-5-thiomorpholin-4'-dioxo- -N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine, as a yellow forming solid (224 mg, 55%). HPLC-MS TR = 3.01 min (UV 254 nm, 5 min method); mass calculated for formula C3IH46BrN7O4SSi2 747.2, observed LCMS m/z 748.0 (M+H).
Part D:
6-(1-ethoxyylnylV3-(1-phenyl-1 H-pyrazoi-4-yl)-5-thiomorpholin-4'"dioxo-N,N-bis(f2-
(trimethylsiiyl)ethoxy)methyl)pyrazoiof1,5-alPyrimidin-7-amine:
A mixture of compound 6-Bromo-3-(l-phenyl-1H-pyrazoI-4-yI)-5-thiomorpholin-4'-dioxo- - N,N-bis((2-(trimethyIsilyl)ethoxy)methyl)pyrazolo[ 1 ,5-a]pyrimidin-7-amine (0.299 mmol, 224 mg), tributyl(l-ethoxyviny[)tin (0.897 mmol, 0.303 mL), Pd(PPh3)4 (0.030 mmol, 34.6 mg) in dioxane (5 mL) was stirred at 100 °C under Ar2 for 16 h. After cooling to rt, the reaction mixture was passed through a short SiCVKF (9:1) plug to removed majority of the Sn species, and then purified by a SiO2 column (0-40% EtOAc/Hexanes, Rf = 0.7 in 50% EtOAc) to afford compound, 6-( L-ethoxyvinyl)-3-(l-phenyl-1H-pyrazol-4-yl)-5-thiomorpholin-4'-dioxo-N,N- bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine as a pale yellow oil (185 mg, 84%). HPLC-MS TR = 3.06 min (UV 254 nm, 5 min method); mass calculated for formula C35H53N7O5SSi2739.3, observed LCMS m/z 622.2 (M-117).
Part E:
1-(7-amino-3-(1-phenyl-1 H-pyrazo!-4-yl)-5-thiomorpholin-4'-dioxo-pyrazolo[1 ,5- alpyrimidin-6-yπethanone:
Compound , 6-(l-ethoxyvinyl)-3-(l-phenyl-1H-pyrazol-4-yl)-5-thiomorpholin-4'-dioxo-N,N- bis((2-(trimethylsi!yl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine, (0.250 mmol, 185 mg) was treated with a mixture of TFAZH2O (2/2 mL) at rt for 30 min. All the volatiles were removed under reduced pressure. The residue was dissolved in 3.0 mL of DMSO and purified by a reverse phase HPLC (H2O/CH3CN, 0.1 % TFA). The product fraction was concentrated to dryness and converted to HCl salt by adding 1 N HCl (aq) solution to its slurry in MeOH, and then concentrated to dryness. Compound, l-(7-amino-3-(l-phenyl-IH-pyrazoi-4-yl)-5- thiomorpholin-4'-dioxo-pyrazolo [1,5-a]pyrimidin-6-yl)ethanone (HCl salt) was obtained as a pale yeliow solid (16.4 mg, 38%). HPLC-MS TR = 1.86 min (UV 254 nm, 5 min method); mass calculated for formula C2iH2!N7O3S 451.1, observed LCMS m/z 452.0 (M+H).
By essentially the same procedures given in Preparative Example above(Scheme-33), compounds given in Table- 1OE can be prepared.
TABLE-IOE
Figure imgf000519_0001
Figure imgf000520_0001
Figure imgf000521_0001
Synthesis of H7-amino-3-(quinolin-3-yl)-5-thiomorpholino-1 ', 1 '-dioxide-pyrazoloM ,5- aipyramidin-6-vθpropan-i -one:
Figure imgf000522_0001
A degassed mixture of 6-bromo-3-(quinolin-3-yl)-5-thiomorpholino- f, l'-dioxide--N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (16 mg, 0,022 mmoL), tributy!(prop-1-ynyl)stannane (22 mg, 0.067 mmoL), Pd(PPh3)4 (2.6 mg, 0.002mmoL) in Dioxane (3 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and filtered through short plug of 10% KFZSiO2. The crude Stille coupling product obtained after concentrating the filtrate was then treated with 1 ; 1 mixture of TFA and H2O at room temperature for 30 min. Concentration and purification by prep-LC afforded i-(7-amino-3- (quinolin-3-yl)-5-thiomorpholino-r, r-dioxide-pyrazolo[1,5-a]pyrimidin-6-yl)propan-1-one. LCMS tR = 3.20 Min ( 10 min run, UV 254nm). Mass calculated for, M+ 450.1, observed LC/MS m/z 451.1 (M+H).
7-(7-amino-6-propionyl-3-(quinolin-3-yl)pyrazolof1.5-aipyrimidin-5- yl)tetrahydroimidazofi ,5-aiPyrazine-1.S^H.SI-D-dione:
Figure imgf000522_0002
By essentially the same procedure given in Preparative Example above, 7-(7-amino-6- propionyl-3-(quinolin-3-yl)pyrazolo[1,5-a]pyrimidin-5-yl)tetrahydroimidazo[l,5-a]pyrazine- 1 ,3(2H,5H)-dione can be prepared from 7-(7-(bis((2-(trimethyIsilyl)ethoxy)methyl)amino)-6- bromo-3-(quinoIin-3-yl)pyrazoIo[1,5-a]pyrimidin-5-yl)tetrahydroimidazo[1,5-a]pyrazine- 1,3(2H,5H)-dione. LCMS tR = 3.14 Min (10 min run, UV 254nm). Mass calculated for, M+ 470.18, observed LC/MS m/z 471.28 (M+H).
Synthesis of 1-(7-amino-3-(αuinolin-3-yl)-5-thiomorpholino-1 '. 1 '-dioxide-pyrazolo[1 ,5- aipyrimidin-6-vO-2-phenylethanone:
Figure imgf000523_0001
By essentially the same procedure given in Preparative Example above, l-(7-amino-3-
(quinolin-3-yl)-5-thiomorpholino-l', r-dioxide-pyrazolo[l,5-a]pyrimidin-6-yl)-2- phenylethanone can be prepared using tributy[(phenyiethynyl)stannane instead of tributyl(prop-1-ynyl)stannane. LCMS IR = 3.74 Min (10 min run, UV 2S4nm)- Mass calculated for, M+ 512.16, observed LC/MS m/z 513.18 (M+H).
Synthesis of 1 -(7-amino-5-(3-methylthiomorpholin-4'dioxo)-3-(1 -phenyl- 1 H-pyrazol-4- yl)pyrazolol"l .5-alpyrimidin-6-vhethanone:
Figure imgf000524_0001
SCHEME-34
Part A:
Synthesis of 3-methyithiomorpholine:
To a solution of KOH (331 mmol, 21.9 g, 85%) in 250 mL of MeOH was added cystamine hydrochloride (158 mmol, 17.9 g), in small portions, followed by a solution of chloroacetone (158 mmol, 12.6 mL) in 100 mL of MeOH at 0-5 °C. After being stirred for 90 min at 0 °C, the reaction was acidified with 350 mL of ! .25 M HCI in MeOH at 0-5 °C and stirred for 1 h. NaBH4 was then added in small portions at below 5 °C and the mixture was stirred for 30 min. The reaction was quenched with 1 N HCl (aq) and filtered. The filtrate was concentrated and then dissolved in H2O and extracted with DCM (*8). The organic layers were discarded and the aqueous phase was carefully neutralized with NaOH first, and then Na2CO3. The aqueous phase was extracted with DCM, dried over Na2SO4 and concentrated to afford compound, 3- methylthiomorpholine (7.69 g, 42%), which slowly turned into a sticky solid and was used without further purification. HPLC-MS TR = 0.19 min (UV 254 nm, 5 min method); mass calculated for formula C5Hi iNS 117.1 , observed LCMS m/z 1 18.2 (M+H).
Part B:
Synthesis of tert-butyl 3-methylthiomorpholine-4-carboxylate: To a solution of compound, 3-methylthiomorpholine (65.6 mmol, 7.69 g) and TEA (131 mmol, 18.3 mL) in THF (200 mL) was added (BoC)2O (98.4 mmol, 21.5 g), followed by DMAP (13.1 mmol, 1.60 g). The resulting reaction mixture was stirred at rt overnight. THF was removed and the residue was dissolved in DCM and directly purified by a SiO2 column (0-15% EtOAc/Hexanes, Rf = 0.55 in 20% EtOAc, KMnO4 staining) to afford compound tert-butyl 3- methylthiomorpholine-4-carboxylate as a white sticky solid (7.31 g, 51%).
Part C:
Synthesis of tert-butyl 3-methylthiomorpholine-4'-dioxo-4-carboxylate: To a solution of compound, tert-butyl 3-methylthiomorpholine-4-carboxylate (33.6 mmol, 7.33 g) in DCM (170 mL) at 0 °C was added mCPBA (101 mmol, 24.0 g) in small portions. The reaction mixture was warmed to rt and stirred for 1 h. DCM was removed and the residue was then partitioned between EtOAc and NaHCO3 (aq). The aqueous layer was extracted with EtOAc (*3). The combined organic layers were washed with brine, dried over Na2SO^ and concentrated. The crude product was purified by a SiO2 column (0-50% EtOAc/Hexanes, Rf = 0.6 in 50% EtOAc) to afford compound, tert-butyl 3-methyIthiomorpholine-4'-dioxo-4- carboxylate as a white solid (6.51 g, 78%).
Part D: Synthesis of 3-methylthiomorpholine-4'-dioxide:
Compound tert-butyl 3-methylthiomorpholine-4'-dioxo-4-carboxylate (20.1 mmol, 5.00 g) was treated with TFA/DCM (25/25 mL) at rt for 1 h. The reaction mixture was evaporated to afford compound, 3-methylthiomorpholine-4' -dioxide as a white solid (TFA salt MW:263.23, 5.18 g, 98%), which was used without further purification. HPLC-MS TR = 0.18 min (UV 254 nm, 5 min method); mass calculated for formula C5H1 1NO2S 149.1, observed LCMS m/z 150.1 (M+H).
Part E:
Synthesis of 5-(3-methylthiomorpholin-4'-dioxo)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoio[1 ,5-a1pyrimidin-7-amine:
A mixture of compound, 3-methylthiomorpholine-4' -dioxide (1.00 mmol, 263 mg), 5-chloro- N,N-bis[(2-trimethylsilylethoxy)methyl]-pyrazolo[1,5-a]pyrimidin-7-amine (0.667 mmol, 286 mg, preparation was reported previously), Pd(OAc)2 and CS2CO3 in dioxane (3 mL) was heated at 1 10 °C for 24 h. The reaction mixture was diluted with EtOAc and washed with H2O and brine, dried over Na2SO4, and concentrated. The crude product was purified by a SiO2 column (0-40% EtOAc/Hexanes, Rf = 0.5 in 50% EtOAc) to afford compound, 5-(3- methylthiomorpholin-4'-dioxo)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazoio[l,5- a]pyrimidin-7-amine, as a colorless oil (145 mg, 40%). HPLC-MS TR = 2.75 min (UV 254 nm, 5 min method); mass calculated for formula C23H43NjO4SSi2 541.3, observed LCMS m/z 542.2 (M+H).
Part F:
Synthesis of 3-iodo-5-(3-methylthiomorpholin-4'-dioxo)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazoio[1 ,5-a|pyrimidin-7-amine:
To a solution of compound, 5-(3-methylthiomorpholin-4'-dioxo)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (0.268 mmol, 145 mg) in CH3CN (5 mL) was added NIS (0.294 mmol, 66.2 mg). The resulting solution was stirred at rt for 30 min. The reaction mixture was evaporated and directly purified by a SiO2 column (0- 15% EtOAc/Hexanes, Rf = 0.55 in 20% EtOAc) to afford compound, 3-iodo-5-(3- methylthiomorpholin-4'-dioxo)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazoIo[1,5- a]pyrimidin-7-amine, as a pale yellow oil (161 mg, 90.0%). HPLC-MS TR = 7.29 min (UV 254 nm, 10 min method); mass calculated for formula C23H42INgO4SSi2 667.2, observed LCMS m/z 668.0 (M+H).
Part G:
Synthesis of 5-(3-methylthiomorphoiin-4'dioxo)-3-(1 -phenyl-1 H-pyrazol-4-yl)-NTN- bisf(2-(trimethylsilyl)ethoxy)methyl)pyrazolori .5-a]pyrimidin-7-amine:
A mixture of compound, 3-iodo-5-(3-memylthiomorpholin-4'-dioxo)-N,N-bis((2- (trimethylsilyl)ethoxy)methyf)pyrazolo[ l,5-a]pyrimidin-7-amine (0.428 mmol, 286 mg), 1- phenyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)»1H-pyrazole (0.557 mmol, 150 mg) , PdCl2(dρpf)CH2Cl2, and K3PO4 in dioxane/H2O (4/0.4 mL) was degassed and then heated at 100 °C for 16 h. After cooling to r.t., the reaction mixture was diluted with EtOAc and filtered. The filtrate was concentrated and purified by a SiO2 column (0-40% EtOAc/Hexanes, Rf = 0.6 in 50% EtOAc/Hexanes) to afford compound, 5-(3-methyIthiomorpholin-4'-dioxo>3-(l- phenyl- 1H-pyrazol-4-yl)-N,N-bis((2-(trimethylsilyi)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin- 7-amine, as a pale yellow oil (142 mg, 48%). HPLC-MS TR = 2.92 min (UV 254 nm, 5 min method); mass calculated for formula C32H49N7O4SSi2 683.3, observed LCMS m/z 684.2 (M+H).
Part H:
1-(7-amino-5-(3-methylthiomorpholin-4'dioxo)-3-(1-phenyl-1 H-pyrazol-4- yl)pyrazolo[1 ,5-aipyrimidin-6-yl)ethanone:
compound, 5-(3-methylthiomorpholin-4'-dioxo)-3-(l-phenyl-1H-pyrazol-4-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoIo[1,5-a]pyrimidin-7-amine, bromination with NBS, foliwed by on stiile reaction with Vinyl triflate and treatment with 4N HCl in dioxane as described in the above example (scheme-???) resulted in a compound, 5-(3- methylthiomorpholin-4'-dioxo)-3-(l-phenyl-1H-pyrazoI-4-yl)pyrazolo[1,5-a]pyrimidin-7- amine as HCl salt. Evaporation of dioxane and lyophilization of the product resulted in 5-(3- methylthiomorpholin-4'-dioxo)-3-( l-phenyl-1H-pyrazol-4-yl) pyrazolo [1, 5-a] pyrimidin-7- amine as white powder. HPLC-MS TR = 4.93 min (UV 254 nm, 10 min method); mass calculated for formula C32H49N7O4S 465.16, observed LCMS m/z 466.10 (M+H).
Synthesis of 6-bromo-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyraπ-4-yl-1 ', V- dioxide)- methyQpyrazolori ,5-aipyrimidin-7-amine
Figure imgf000528_0001
SCHEME-35
Part A:
3-oxo-3-(tetrahydro-2H-thiopyran-4-yl)propanenitriie:
A solution of CH3CN (48,7 mrnol, 2.54 mL) in 10 mL of THF was added dropwise to a solution of "BuLi (48.7 mniol) in 15 mL of THF at -78 °C. After stirring for 1 h at -78 °C, a solution of methyl tetrahydro-2H-thiopyran-4-carboxylate (24.3 mmol, 3.90 g) in 10 mL of TΗF was added dropwise and the resulting reaction mixture was stirred at -78 °C for 1 h, then slowly warmed to -40 °C in 2 h. The reaction was quench with 1 N HCl (pΗ<l). THF was removed and the residue was extracted with EtOAC, washed with brine, dried over Na2SO4, and concentrated. The crude product was purified by a SiO2 column (0-50% EtOAc/Hexanes, Rf = 0.45 in 50% EtOAc) to afford compound 3-oxo-3-(tetrahydro-2H-thiopyran-4- yl)propanenitrile (3.60 g, 87.5%) as a light brownish oil. HPLC-MS TR = 1.15 min (UV 254 nm, 5 min method); mass calculated for formula CeHnNOS 169.1 observed LCMS m/z 170.1 (M+H).
Part B: 5-(tetrahydro-2H-thiopyran-4-yl)-N.N-bis((2-(trimethylsilvhethoxy)methynpyrazolori .5- aipyrimidin-7-aminβ: A mixture of 3-aminopyrazole (19.3 mmol, 1.61 g) and compound 3-oxo-3-(tetrahydro-2H- thiopyran-4-yl) propanenitrile (21.3 mmol, 3.60 g) in acetic acid (40 mL) was heated at 100 °C in a sealed tube overnight. After cooling to rt, all the volatiles were removed under reduced pressure to afford a light brownish oil. This brownish oil was dissolve in DCM (60 mL), and then SEMCI (67.6 mmoi, 11.9 mL) and DIPEA (135 mmol, 23.5 mL) were added. The resulting reaction mixture was stirred at 45 °C for 1 h. After cooling to rt, all the volatiles were removed under reduced pressure. The residue was diluted with EtOAc (300 mL), filtered and washed with EtOAc. The filtrate was concentrated and the crude product was purified by a SiO2 column (0-30%) to afford compound, 5-(tetrahydro-2H-thiopyran-4-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine as a light brownish oil (9.23 g, 97%). HPLC-MS TR = 3.09 min (UV 254 nm, 5 min method); mass calculated for formula C23H42N4O2SSi2494.3, observed LCMS m/z 495.1 (M+H).
Part C: 5-(tetrahydro-2H-thiopyran-4-yl-1 '. 1 '-dioxide)-)-N,N-bis(f2- ftrimethylsiiyl)ethoxy)methyl)pyrazolof1 ,5-alpyrimidin-7-amine: To a solution of compound, 5-(tetrahydro-2H-thiopyran-4-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methy0pyrazolo[1,5-a]pyrimidin-7-amine (18.7 mmol, 9.23 g) in DCM (100 mL) was added mCPBA and stirred at rt for 1 h. The reaction was quenched with Na2S2O3 (aq.) and diluted with DCM (100 mL), The separated organic layer was washed with NaHCO3 (2*) and brine, dried over Na2SO4, and concentrated to afford crude compound, 5- (tetrahydro-2H-thiopyran-4-yl-l ', 1 '-dioxide)-)-N,N-bis((2-
(trimethylsilyi)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-arnine as a purple oil, which was used without further purification. HPLC-MS TR = 2.72 min (UV 254 nm, 5 min method); mass calculated for formula C23H42N4O4SSi2 526.2, observed LCMS m/z 527.2 (M+H).
Part D:
3-iodo-5-(tetrahydro-2H-thioDyran-4-yl-1.1-dioxideVN.N-bis(f2- (trimethylsilyl)ethoxy)methyl)pyrazolof1 ,5-aipyrimidin-7-amine: To a solution of all crude compound, 5-(tetrahydro-2H-thiopyran-4-yl- 1 ', 1 '-dioxide)-)-N,N- bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine from Part C (18.7 mmol) in CH3CN (100 mL) was added NIS (20.6 mmol, 4.63 g). The resulting solution was stirred at it for 30 min. TLC (50% EtOAc/Hexanes) indicated complete conversion to a new spot (Rf= 0.5, less polar than SM Rf = 0.3). The reaction was quenched with Na2S2O3. CH3CN was removed under reduced pressure. The residue was dissolved in DCM, washed with H2O and brine, dried over Na2SO4, and concentrated. The crude product was purified by a SiO2 column (0-50% EtO Ac/Hex anes) to afford compound, 3-iodo-5-(tetrahydro-2H-thiopyran-4- yl- 1 , l-dioxide)-N,N-bis((2-(trimethyIsilyI)ethoxy)methyl)pyrazoIo[ 1 ,5-a]pyrimidin-7-amine: as a pale yellow oil (6.82 g, 56%).
Part E: 3-(5-phenylpyridin-2-yl)-5-(tetrahydro-2H-thiopyran-4-yl-1 ,1 -dioxide)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoiof1 ,5-alpyrimidin-7-amine:
To a solution of 2-bromo-5-phenylpyridine (0.400 mmol, 93.6 mg) and triisopropyl borate (0.440 mmol, 0.102 mL) in a mixed solvent of toluene/THF ( 1.6/0.4 mL) was added dropwise "BuLi (0.440 mmol) at -78 °C, then kept at -78 °C for 30 min (sticky mixture, stopped stirring after 5 min). The reaction mixture was allowed to warm to rt and stirred for 4 h. All the volatiles were removed under reduced pressure from rt to 90 °C. The thus formed crude lithium borate was used without further purification. A mixture of the crude lithium borate (0.400 mmol), compound, 3-iodo-5-(tetrahydro-2H-fhiopyran-4-yl-I,l-dioxide)-N,N-bis((2- (trimethytsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine (0.267 mmol, 174 mg),
PdCl2(dppf) CH2Cl2 (0.027 mmol, 22.0 mg), and KF (0.800 mmol, 46.5 mg) in dioxane (2 mL) was degassed and then heated at 90 °C for 16 h. The reaction mixture was diluted with EtOAc and filtered. The filtrate was concentrated and purified by a SiO2 column (0-40% EtOAc/Hexanes, Rf = 0.5 in 50% EtO Ac/Hex anes) to afford compound, 3-(5-phenylpyridin-2- yl)-5-(tetrahydro-2H-thiopyran-4-yl- 1 , 1 -dioxide)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazo[o[l ,5-a]pyrimidin-7-amine as a pale yellow solid (32 mg, 18%). HPLC-MS TR = 2.62 min (UV 254 nm, 5 min method); mass calculated for formula C34H49N5O4SSi2 679.3, observed LCMS m/z 680.2 (M+H).
Part- F:
6-bromo-3-(6-phenylpyridin-3-vh-5-(tetrahydro-2H-thiopyran-4-yl-1 '. 1 '-dioxideVN.N- bisf(2-(trimethylsilyl)ethoxy)methyl)pyrazolori ,5-a1pyrimidin-7-amine: NBS (176 mg, 0.99 mmoL) was added to a solution of 3-(6-phenyipyridin-3-yl)-5-(tetrahydro- 2H-thiopyran-4-yl-l', r-dioxide)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5- a]pyrimidin-7-amine (679 mg, 0.94 mmoL) in CH3CN (10 mL). The mixture was stirred at room temperature for Ih. Concentration and purification by column chromatography afforded 6-bromo-3-(6-pheny lpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl- 1 ', 1 '-dioxide)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidirι-7-amine. LCMS .R = 2.87 Min (5 min run, UV 254nm). Mass calculated for, MH- 757, observed LC/MS m/z 758.2 (M+H). Part G:
6-bromo-3-(6-phenylpyridin-3-yl)-5-(tΘtrahydro-2H-thiopyran-4-yl-1 ', 1 -dioxide)- methyl)pyrazolof1 ,5-a]pyrimidin-7-amine:
6-bromo-3-(6-phenyIpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-l', l'-dioxide)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine was treated with 4NHC1 in dioxane at room temperature for 1 hr and reaction evaporated to dry ness and lyophilized under vacuo to give product in powder form, 6-bromo-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H- thiopyran-4-yl-T, 1 '-dioxide)- methyl)pyrazolo[i ,5-a]pyrimidin-7-amine as HCl salt. LCMS IR = 1.27 Min (5 min run, UV 254nm). Mass calculated for, M+ 497.10, observed LC/MS m/z 498.20 (M+H).
By essentially the same procedure given in Preparative above(Scheme-35), compounds 10.1 16 to 10.133 given in the table (10-F) can be prepared.
Figure imgf000531_0001
Figure imgf000532_0001
Figure imgf000533_0001
Figure imgf000534_0002
Synthesis of 3-(6-phenylpyridin-3-vO-5-(tetrahydro-2H-thiopyran-4--yl-1 M '-dioxide)- N,N-bis((2-(trimethylsilyl)ethoxy)methyl)-6-ylnylpyrazoio[1 ,5-alpyrimidin-7-amine
Figure imgf000534_0001
SCHEME-36 Part A
3-(6-phenylpyridin-3-vn-5-(tetrahydro-2H-thiopyran-4-yl-1M'-dioxide)-N,N-bis((2- (trimethyisilyl)ethoxy)methyl)-6-ylnylpyrazolori ,5-a1pyrimidin-7-amine:
A degassed mixture of 6-bromo-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl- r,r-dioxide)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (203 mg, 0.27mmol), Pd(PPh3)4 (31 mg, 0.027 mmoL), tributyl(vinyl)stannane (255 mg, 0.80 mmoL) in CH3CN (6 mL) was heated at 150°C under microwave condition for 60 min. The reaction mixture was cooled to room temperature, filtered through 9: 1 Siθ2:KF plug and concentrated in vacuo. Purification by column chromatography afforded 3-(6-phenylpyridin-3- yl)-5-(tetrahydro-2H-thiopyran-4-yl- 1 ' , r-dioxide)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)- 6-vinylpyrazolo[1,5-a]pyrimidin-7-amine: LCMS tR = 2.82 Min (5 min run, UV 254nm). Mass calculated for, M+ 705.3, observed LC/MS m/z 706.3 (M+H).
Part B
7-(bisf(2-(trimethyisiiyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3-ylV5-(tetrahydro- 2H-thiopyran-4-yl-1 M '-dioxide)pyrazolori ,5-aipyrirnidine-6-carbaidehyde:
To 3-(6-phenylpyridin-3-y l)-5-(tetrahydro-2H-thiopyran-4-yl- 1 ' , 1' -dioxide)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)-6-vinylpyrazolo[1,5-a]pyrimidin-7-amine (189 mg, 0.26 mmol) in 1 , 4-doixane (3 mL) was added 2.5 wt% OsO4 in 1 , 4-dioxane ( 168 uL, 0.013 mmol), 2,6- kitidine (265 uL, 2.68 mmol) and H2O (1 mL) and the resulting mixture was stirred at room temperature for 20 minutes. NaIO4 (287 mg, 1.34 mmol) was then added and stirring at room temperature continued for 4 days. Saturated Na2S2O3 solution (5 mL) was added and the mixture stirred for 10 minutes. Organic s were then extracted with CH2Cl2 (2 x), dried (Na2SO4) and concentrated in vacuo to 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-r,r-dioxide)pyrazolo[l,5-a]pyrimidine- 6-carbaldehyde, which was used without further purification: LCMS tR = 2.87 Min (5 min run, UV 254nm). Mass calculated for, M+ 707.2, observed LC/MS m/z 708.3 (M+H).
Part C 7-amino-3-(6-pheπylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-1 ', 1 '- dioxide)pyrazoiol"1.5-aipyrimidine-6-carbaldehyde:
The crude 7-(bis((2-(ti-imethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3-yi)-5- (tetrahy dro-2H-thiopyran-4-y 1 - 1 ' , 1 ' -dio xide)py razo io [ 1 ,5- a]pyr i midine-6-c arbaldehy de (150 mg, 0.21 ramoL) was treated with 50% TFA in H2O (4 mL) until the disappearance of starting material in LCMS. Concentration and purification by prep-LC afforded 7-amino-3-(6- phenyIpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-l', l'-dioxide)pyrazolo[l ,5- a]pyrimidine-6-carbaldehyde, LCMS tR = 3. !0 Min (10 min run, UV 2s4nm)- Mass calculated for, M+ 447.1 , observed LC/MS m/z 448.1 (M+H).
By essentially the same procedure given in Preparative Example(Scheme-36) compounds 10.134- 10.137 ( Table-10-G) can be prepared.
TABLE- 10 G
Figure imgf000536_0001
Figure imgf000537_0002
Synthesis of i -fy-amino-3-(β-phenylpyridin-3-yl)-5-(tetrahydro^H-thiopyran-4-yl-1 ', 1 '- dioxide)pyrazolo[1 ^-alpyrimidin-6-yl)propan-i -one:
Figure imgf000537_0001
By essentially the same procedure given in Preparative Example above(Scheme-36), l-(7- amino-3-(6-phenylpyridm-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl- 1 ', l'-dioxide)pyrazolo[1,5- a]pyrimidin-6-yl)propan-1-one can be prepared from 6-bromo-3-(6-phenylpyridin-3-yl)-5- (tetrahydro-2H-thiopyran-4-yl-r, 1 '-dioxide)-N,N-bis((2 trimethylsilyl) ethoxy) methyl)pyrazolo[1,5-a]pyrimidin-7-amine. LCMS tR = 3.13 Min ( 10 min run, UV 254nm). Mass calculated for, M+ 449.15, observed LC/MS m/z 450.40 (M+H). Synthesis of 1 -(7-amino-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-1 ', 1 '- dioxide)pyrazolo[1.5-aipyrimidin-6- yl)ethanone:
Figure imgf000538_0001
A degassed mixture of 6-bromo-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl- r,r-dioxide)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7-amine (203 mg, 0.27mmol), Pd(PPh3)4 (31 mg, 0.027 mmoL), tributyl(vinyl)stannane (255 mg, 0.80 mmoL) in CH3CN (6 mL) was heated at 15O°C under microwave condition for 60 min. The reaction mixture was cooled to room temperature, filtered through 9:1 SiO2:KF plug and concentrated in vacuo. The crude β-Cl-ethoxyvinyO-3-Cό-phenylpyridin-3-yl)-5-Ctetrahydro- 2H-thiopyran-4-yl-r,r-dioxide)-N,N-bis((2-(trimethylsUyl)ethoxy)methyI)pyrazolo[1,5- a]pyrimidϊn-7-aminewas treated with 50% TFA in H2O (2 mL) for Ih. Concentration and purification afforded l-(7-amino-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yi- Γ,1' -dioxide)pyrazolo[1,5-a]pyrimidin-6-yl)ethanone: LCMS IR = 3.18 Min (10 min run, UV 254nm). Mass calculated for, M+ 461.1, observed LC/MS m/z 462.1 (M+H).
6-(methylsulfonvh-3-(Quinolin-3-yi)-5-thiomorpholino-1 M '-dioxide-pyrazoioH ,5- aipyrimidin-7-amine:
Figure imgf000538_0002
A mixture of 6-bromo-3-(quinolin-3-yl)-5-thiomorpholino-r,r-dioxide-N,N-bis((2- (trimethyIsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (0.06J mmol, 45 mg), CuI (0.49 mmol, 94 mg) and NaSO2Me (0.49 mmol, 50 mg) in NMP (2mL) was heated at 12O°C for Ih and then 13O°C for Ih under microwave condition. Purification by prep-LC afforded 6- (methylsulfonyl)-3-(quinolin-3-yl)-5-thiomorpholino--r, l'-dioxide-pyrazolo [1,5- a]pyrimidin-7-amine: LCMS tR = 2.94 Min (10 min run, UV 254nm)- Mass calculated for, M+ 472.0, observed LC/MS m/z 473.0 (M+H).
Figure imgf000539_0001
A degassed mixture of 6-bromo-5-thiomorpholinodioxo-3-(quinoIin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyra2olo[1,5-a]pyrimidin-7-amine (35 mg, 0.051mmoL), Bu3SnCN (32.3 mg, 0.10 mmoL), Pd[P(t-Bu)3]2 (5.2 mg, 0.010 mmoL) in Dioxane (3 mJL) was heated at 100°C overnight. The mixture was cooled to room temperature and the solvent was evaporated. Purification by column chromatography afforded 7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-5-thiomorpholinodioxo-3-(quinoiin-3-yl)pyrazolo[l,5- a]pyrimidine-6-carbonitrile. LCMS tβ = 2.76 Min (5 min run, UV 254nm). Mass calculated for, M+ 678.28, observed LC/MS m/z 679.20 (M+H).
A mixture of 7-(bis((2-(trimethylsilyL)ethoxy)methyl)araino)-5-thiomorpholinodioxo-3- (quinoiin-3-yl)pyrazolo[i,5-a]pyrimidine-6-carbonitrile (5 mg), EtOH (ImL) and 3N HCl (ImL) was heated at 6O°C until LCMS indicated complete reaction. The mixture was cooled to room temperature and the solvent was evaporated. Purification by prep-LC afforded 7-amino- 5-thiomorpholinodioxo-3-(quinolin-3-yl) pyrazolo [1, 5-a] pyrimidine-6-carbonitrile. LCMS tR = 3.12 Min (10 min run, UV 254nm)- Mass calculated for, M+ 418.12, observed LC/MS m/z 419.2 (M+H).
Following the examples described above, the compounds (Scheme-36)) given in the Table- 10- H can be synthesized
TABLE-IO-H
Figure imgf000540_0001
Synthesis of 7-amino-3-(Quinolin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-1 M'- dioxide)pyrazoiof1 ,5-aipyrimidine-6-carboxamide:
Figure imgf000541_0001
The 7-amino-3-(quinoUn-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl- 1', 1 '-dioxide)pyrazolo[ 1 ,5- a]pyrimidine-6-carbonitriiewas stirred in cone. H2SO4(ImL) at 6O°C until LCMS indicated complete conversion. The reaction was quenched with ice and the pH was adjusted to ~ 8 by 6 N NaOH. The mixture was extratce by 15% j-PrOH/CH2Cl2 (x3). The combined organic layers were dried with Na2SO^ Evaporation of solvent afforded the crude displacement compound. Purification by prep-LC afforded 7-amino-3-(quinolin-3-yl)-5-(tetrahydro-2H- thiopyran-4-yl- l',r-dioxide)pyrazolo[i,5-a]pyrimidine-6-carboxamide. LCMS tR = 2.27 Min (10 min run, UV 254nm). Mass calculated for, M+ 436.13, observed LC/MS m/z 437.23 (M+H).
Synthesis of 7-amino-3-(quinolin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl)pyrazolo[1 ,5- a]pyrimidine-6-carboxamide:
Figure imgf000541_0002
By essentially the same procedure given in Preparative above, 7-amino-3-(quinolin-3-yl)-5- (tetrahydro-2H-thiopyran-4-yl) pyrazolo [1,5-a]pyrimidine-6-carboxamide can be prepared. LCMS tR = 3.21 Min (10 min run, UV 2s4!im). Mass calculated for, M+ 405.13, observed LC/MS m/z 406.15 (M+H).
Synthesis of 1-(7-amino-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-1M '- dioxide)pyrazolo[1 ,5-a]pyrimidin-6-yl)ethanone oxime:
Figure imgf000541_0003
A mixture of l-(7-amino-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-r, 1'- dioxide)pyrazolo [1,5-a]pyrimidin-6-yl)ethanone (4mg) and 50% NH2OH (120 uL) in NMP (ImL) was heated at lOOoC for 30 min under microwave condition. Purification by prep-LC afforded t-(7-amino-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-r,r- dioxide)pyrazolo[ 1 ,5-a]pyrimidin-6-yl)ethanone oxime: LCMS tR = 3.04 Min ( 10 min run, UV 254nm). Mass calculated for, M+ 476.16, observed LC/MS m/z 477.59 (M+H).
Synthesis of 1-amino-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyraπ-4-yl--1', 1'- dioxide)pyrazoio[1 ,5-a]pyrimidine-6-carbaldehyde oxirne:
Figure imgf000542_0001
By essentially the same procedure given in Preparative Example given above l-amino-3-(6- phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl--r, l'-dioxide) pyrazolo [1,5- a]pyrimidine-6-carbaldehyde oximecan be prepared. LCMS tR = 3.15 Min (10 min run, UV 254nm)- Mass calculated for, M+ 462.14, observed LC/MS m/z 463.57 (M+H).
Synthesis of 1-(7-amino-3-(αuinoiin-3-vO-5~thiomorpholinopyrazoio-1', 1'-dioxide-f1.5- aipyrimidin-6-vhethanol:
Figure imgf000542_0002
SCHEME-37
NaBH4 (10 mg) was added to a solution of l-(7-amino-3-(quinolm-3-yl)-5-thiomorpholino-r, l'-dioxide-pyrazoio [1, 5-a]pyrimidin-6-yl)ethanone (10 mg, 0.023 mmoL) in MeOH (1 mL). After stirring at room temperature for Ih, the mixture was concentrated to dry. Purification by prep-TLC afforded l-(7-amino-3-(quinolin-3-yI)-5-thiomorpholinopyrazolo-r, l '-dioxide- [1,5-a]pyrimidin-6-yl)ethanol: LCMS tR = 2.44 Min (5 min run, UV 254nm)- Mass calculated for, M+ 438.1, observed LC/MS m/z 439.1 (M+H).
By essentially the same procedure given in Preparative Example described above(Scheme-37), following compounds 10.142-10.148 in table-10-I can be prepared.
TABLE-IQ-I
Figure imgf000543_0001
Figure imgf000544_0001
Synthesis of 3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl--1 M '-dioxide)-6- (thiophen-2-yl)pyrazoiof1 ,5-aipyrimidin-7-amine:
Figure imgf000545_0001
By essentially the same procedure given in Preparative Example described above(Scheme-36), 3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl- 1 ' , l'-dioxide)-6-(thiophen-2- yl)pyrazolo[i,5-a]pyrimidin-7-amine be prepared. LCMS tR = 4.18 Min (10 min run, UV 254πm). Mass calculated for, M+ 501.12, observed LC/MS m/z 502.0 (M+H).
By essentially the same procedure given in Preparative Example above(36), compounds in table 10-J ( 10.149 -10.151) can be prepared.
TABLE-10-.T
Figure imgf000545_0002
Figure imgf000546_0002
Synthesis of β-cvciopropyl-3-(β-phenylpyridin-3-vh-5-ftetrahydro^H-thiopyran-Φyl-- 1 M '-dioxide)pyrazolo[1 ,5-aipyrimidin-7~amine:
Figure imgf000546_0001
SCHEME-38
A degassed mixture of 6-bronio-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl- l',i '-dioxide)-N,N-bis((2-(tπmethylsilyl)ethoxy)methyl)pyrazoio[1,5-a]pyrimidin-7-amine (53 mg, 0.07mmol), Pd(OAc)2 (3.1 mg, 0.014 mmoL), P(C6H11)3 (7.9 mg, 0.028 mmoL), 2- cyclopropyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (23.5 mg, 0.14 mmoL), K3PO4 (53 mg, 0.25 mmoL) in Toluene (0.4 mL) and H2O (0.02 mL)was heated at 10O°C overnight. The reaction mixture was cooled to room temperature, filtered through celite and concentrated in vacuo. The crude coupling product was treated with 50% TFA in H2O (4 mL) for Ih. The reaction mixture was concentrated in vacuo. Concentration and purification afforded 6- cyclopropyl-3-(6-phenylpyridin-3-yI)-5-(tetrahydro-2H-thiopyran-4-yl-- IM'- dioxide)pyrazolo[1,5-a]pyrimidin-7-amine: LCMS tβ = 3.49 Min (10 min run, UV 254nm). Mass calculated for, M+ 459.17, observed LC/MS m/z 460.22 (M+H).
By essentially the same procedure given in Preparative Example above(Scheme-38), compounds 10.152 - 10.154 can be prepared are given in Table-10-K
TABLE-IO-K
Figure imgf000547_0001
Figure imgf000548_0001
Scheme-39
Part A:
(7--(bisf(2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3-yl)-5-(tetrahydro- 2H-thiopyran~4-yl-1',1 '-dioxide) pyrazolo[1 ,5-alpyrimidin-6-yl)methyl methanesulfonate:
At O°C, Et3N (41.7 uL, 0.29 mmol) and MsCl (17.4 uL, 0.22 mmol) were added to a solution of (7-(bis((2-(trimethylsilyl)ethoxy)methyl)aniino)-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H- thiopyran-4-yl-r,r-dioxide)pyrazoIo[l,5-a]pyrimidin-6-yl)methanol (53 tng, 0.75 mmol)in THF (3mL). After stirring at room temperature for Ih, the compound, (7-(bis((2- (trimethylsilyl)ethoxy)raethyl)amino)-3-(6-phenylpyridin-3-yl)-5-(teti-ahydro-2H-thiopyran-4- y I- 1',Γ -dioxide)pyrazolo[l,5-a]pyrimidin-6-yl)methyl methanesulfonate was evenly transferred to three vials. Part B:
6-(aminomethyπ-3-(6-phenylpyriciin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-1 M '-dioxide
)pyrazoio[1 ,5-alpyrimidin-7-amine:
7N NH3 in MeOH (ImL) added was added to vial I. 6-(aminomethyl)-3-(6-phenyipyridin-3- yl)-5-(tetrahydro-2H-thiopyran-4-yl-l', l'-dioxide) pyrazolo[l,5-a]pyrimidin-7-amine was obtained after concentration and deprotection by treating with 50% TFA in H2O and then prep- LC purification to give the product, 6-(aminomethyI)-3-(6-phenylpyridin-3-yl)-5-(tetrahydro- 2H-thiopyran-4-yl-l', l'-dioxide) pyrazolo[1,5-a]pyrimidin-7-amine. LCMS tR = 1.97 Min ( 10 min run, UV 254nm). Mass calculated for, M+ 448.16, observed LCMS m/z 449.2 (M+H).
Part C:
6-((methylamino)methyl)-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-1 M '- dioxide) pyrazoloH ,5-a1pyrimidin-7-amine:
2 N MeNH2 in THF (ImL) added was added to vial 2. 6-((methylamino)methyI)-3-(6- phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-r, r-dioxide)pyrazolo[l,5-a]pyrimidin- 7-amine was obtained after concentration and deprotection by treating with 50% TFA in H2O and then prep-LC purification resulted in pure 6-((methylamino)metb.yl)-3-(6-phenyIpyridin-3- yl)-5-(tetrahydro-2H-thiopyran-4-yl-l\t '-dioxide) pyrazolo[1,5-a]pyrimidin-7-amine as TFA salt. LCMS tR = 2.12 Min (10 min run, UV 254nm). Mass calculated for, M+ 462.18, observed LC/MS m/z 463.3 (M+H).
Part D: 6-(methoxynnethyl)-3-(6-phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4- yl)pyrazolori ,5-aipyrimidin-7-amine:
0.5 M NaOMe in MeOH (1 mL) added was added to vial 3. 6-(methoxymethyl)-3-(6- phenylpyridin-3-yl)-5-(tetrahydro-2H-thiopyran-4-yl-l ', 1 '-dioxide)pyrazoio[l ,5-a]pyrimidin- 7-aminewas obtained after concentration and deprotection by treating with 50% TFA in H2O and then prep-LC purification provided pure compound, 6-(methoxymethyl)-3-(6- phenylpyridin-3-y[)-5-(tetrahydro-2H-thiopyran-4-yl- i ', l'-dioxide)pyrazolo[l,5-a]pyrimidin- 7-amine. LCMS tR = 2.66 Min (10 min run, UV 254nm). Mass calculated for, M+ 463.1, observed LC/MS m/z 464.1 (M+H).
By essentially the same procedure given in Scheme-39, following compounds 10.155 to 10.161can be prepared. (Table-10-L)
TABLE-IO-L
Figure imgf000550_0001
Figure imgf000551_0001
Synthesis of tert-butyl 2-(7-(bis((2-(trimethyisilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5-yl)-5-methoxy-1 H-indole-1 -carboxylate:
Figure imgf000552_0001
Htert-butoxycarbonyl)-5-methoxy-1H-indol-Z-ylboronic acid (0.24 mmol, 70 mg), K3PO4 (0.36 mmoi, 77 mg), and PdCl2(dppf>CH2Cl2 (0.012 mmol, 9.8 mg) were added to a solution of 5-chloro-3-(6-phenyIpyridin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1,5- a]pyrimidin-7-amine (0.12 mmol, 72 mg) in dioxane (4 mL) and H2O (0.5 mL). The resulting solution was stirred at 90° C under argon overnight. The mixture was diluted with H2O and then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation and purification by column chromatography afforded tert- butyl 2-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3-yl)pyrazolo[1,5- a]pyrimidJn-5-yl)-5-methoxy- 1H-indoie-1-carboxylate, LCMS tR = 3.48 Min (5 min run, UV 254nm). Mass calculated for, M+ 792.3, observed LC/MS m/z 793.2 (M+H).
6-bromo-5-(5-methoxy-1 H-indol-2-yl)-3-(6-pheπylpyridin-3-yl)pyrazolof1 ,5-a1pyrimidin- 7-amine:
Figure imgf000552_0002
By essentially the same procedure given in Preparative Examples above, 6-bromo-5-(5- methoxy- 1 H-indol-2-yl)-3-(6-pheny lpyridin-3-yl)pyrazolo[ 1 ,5-a]pyrimidin-7-amine can be prepared. LCMS IR = 4.59 Min (10 min run, UV 254nm)- Mass calculated for, M+ 510.08, observed LC/MS m/z 5 U.50 (M+H).
Synthesis of 5-chloro-3-(6-phenylpyπdin-3-yl)-N,N-bis((2- (trimethyisilyl)ethoxy)methyl)pyrazolo[1 ,5-aipyrimidin-7-amine:
Figure imgf000553_0001
2-phenyl-5-(4,4,5,5-tetraraethyl-1,3,2-dioxaboroian-2-yl)pyridine (2.38 mmol, 675 mg), K3PO4 (5.96 mmol, 1264 mg), and PdCl2(dppfyCH2Cl2 (0.20 mmol, 162 mg) were added to a solution of 5-chloro-3-iodo-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[I,5-a]pyrimidin-7- amine (1.98 mmol, UOl mg) in dioxane (18 mL) and H2O (3 mL). The resulting solution was stirred at 70° C under argon overnight. The mixture was diluted with H2O and then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation and purification by column chromatography afforded 5-chloro-3-(6- phenyipyridin-3-yl)-N,N-bis((2 trimethylsilyl)ethoxy)methyl)pyraεoio[l ,5-a]pyrimidin-7- amine: LCMS IR = 3.36 Min (5 min run, UV 254nm)- Mass calculated for, M+ 581.2, observed LC/MS m/z 582.2 (M+H).
Synthesis of tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)pyrazoiori .5-alPyrimidin-5-yloxy)piperidine-1-carboxyiate:
NaH (120 mg, 3 mmoL) was added to a solution of tert-butyl 4-hydroxypiperidine-1- carboxylate (522 mg, 2.60 mmoL) in THF (8 mL). After stirring at room temperature for 5 min, 5-chloro-3-(6-phenyipyridin-3-yl)-N,N-bis((2-
(trimethylsiIyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (755 mg, 1.30 mmoL) in THF (4 mL) was added dropwise. The mixture was heated under microwave condition at 100°C for 30 min, diluted with H2O and then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation and purification by column chromatography afforded tert-butyl 4-(7-(bis((2-(trimetrtylsilyl)ethoxy)methyt)amino)- 3-(6-phenylpyridin-3-yl)pyrazolo[I,5-a]pyrimidin-5-yloxy)piperidine-1-carboxylate: LCMS tR = 3.53 Min (5 min run, UV 254«m). Mass calculated for, M+ 746.4, observed LC/MS m/z 747.2 (M+H).
Synthesis of tert-butyl 4-(7-(bisf(2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6- phenylpyridin-3-yl)pyrazoloπ .5-aipyrimidin-5-yloxy)pipericiine-1-carboxylate:
Figure imgf000554_0001
NBS (246 mg, 1.38 mmoL) was added to a solution of tert-butyl 4-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5- yloxy)piperidine-1-carboxylate (1022 mg, 1.38 mmoL) in CH3CN (15 mL). After stirring at room temperature for 30 min, the mixture was concentrated in vacuo. Purification by column chromatography afforded tert-butyl 4-(7-(bis((2-(trimethy!siiyi)ethoxy)methyl)amino)-6- bromo-3-(6-phenylpyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5-yloxy)piperidine-1-carboxyIate: LCMS tR = 3.59 Min (5 min run, UV 254nm). Mass calculated for, M+ 824.3, observed LC/MS m/z 825.1 (M+H).
The tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-phenylpyridin- 3-yl)pyrazolo(1,5-a]pyrimidin-5-yloxy)piperidine-1-carboxylate (143 mg, OJ 7 mmoL) was treated with 50% TFA in H2O (6 mL) and stirred overnight. The reaction mixture was concentrated in vacuo.
(R)-1 -(4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolof1 ,5-a1pyrimidin-5- y1oxy)piperidin-1 -yl)-2-hydroxypropan-1 -one:
A mixture of the crude 6-bromo-3-(6-phenylpyridin-3-yl)-5-(piperidin-4-yloxy)pyrazolo[1,5- a]pyrimidin-7-amine, D-(-)-iactic acid (18.5 mg, 0.20 mmoL), EDC (78.7 mg, 0.41 mmoL), HOBt (27.7 mg, 0.20 mmoL) and DIEA ( 238 uL, 1.37 mmoL) in DMF ( 4 mL) was stirred at room temperature for 2 hr. Purification with prep-LC provided (R)-1-(4-(7-amino-6-bromo-3- (6-phenylpyridin-3-yl)pyrazolo[l,5-a]pyrimidin-5-yloxy)piperidin-1-yl)-2-hydroxypropan-l- one, LCMS tR = 4.39 Min (10 min run, UV 254nm). Mass calculated for, M+ 513.0, observed LC/MS m/z 514.43 (M+H).
Synthesis of 1 -(7-amino-3-(6-phenylpyridin-3-yl)-5-(piperidin-4-yloxy)pyrazolori .5- alpyrimidin-6-yl)ethaπoπe:
Figure imgf000555_0001
A degassed mixture of tert-butyl 4-(7-(bis((2-(trimethytsilyl)ethoxy)methyl)amino)-6-bromo» 3-Cό-phenylpyridin-3-yl)pyrazolotl^-aJpyrimidin-5-yloxyJpiperidine-1-carboxylate (0.57 mmol, 486 mg), Pd(PPh3)4 (66 mg, 0.056 mmoL), tributyl(l-ethoxyvinyl)stannane (410mg, 1.13 mmoL) in Dioxane (6 mL) was heated at 100°C overnight. The reaction mixture was cooled to room temperature, filtered through 9: 1 Siθ2:KF plug and concentrated in vacuo.
The crude tert-butyl 4-(7-(bts((2-(trimethyIsilyl)ethoxy)methyl)amino)-6-( l-ethoxyvinyl)-3-(6- phenylpyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5-yloxy)piperidine-1-carboxylate was treated with 50% TFA in H2O (6 mL) and stirred overnight. The reaction mixture was concentrated in vacuo. Purification with prep-LC provided l-(7-amino-3-(6-phenylpyridin-3-yl)-5-(piperidin- 4-yloxy) pyrazolo [1,5-a]pyrimidin-6-yl)ethanone: LCMS IR = 1.1 IMin (5 min run, UV 254nm). Mass calculated for, M+ 428.1 , observed LC/MS m/z 429.1 (M+H).
(R)-1-(4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolori ,5-a1pyrimidin-5- y[oxy)piperidin-1 -vO-2-hydroxypropan-1 -one:
A mixture of t-(7-amino-3-(6-phenyJpyridin-3-yl)-5-(piperidm-4-yloxy)pyrazolo[1,5- a]pyrimidin-6-yl)ethanone (21 1 mg, 0.49 mmoL), D-(-)-lactic acid (53.3 mg, 0.59 mmoL), EDC (189.0 mg, 0.99 mmoL), HOBt (132.0 mg, 0.99 mmoL) and DEA ( 514 uL, 2.96 mmoL) in DMF (5 mL) was stirred at room temperature for 2 hr. Purification with prep-LC provided (R)-I -(4-(6-acetyl-7-amino-3-(6-phenylpyridin-3-yl)pyrazolo[ 1,5-a]pyrimidin-5- yloxy)piperidin-1-yl)-2-hydroxypropan-1-one: LCMS tR = 3.53 Min (10 min run, UV 254nm). Mass calculated for, M+ 500.21, observed LC/MS m/z 501.62 (M+H).
Figure imgf000556_0001
R = H or F
SCHEME-40
By essentially the same procedure given in Preparative Examples above and scheme-40, compounds 10.162 and 10.163 can be prepared ( Table- 10-M).
TABLE-IO-M
Figure imgf000556_0002
Figure imgf000557_0001
SCHEME-41
By essentially the same procedure given in Preparative Example(Scheme-4l), 10.164 and 10.165 can be prepared.(Table- 10-N)
TABLE-IO-N
Figure imgf000557_0002
Synthesis of tert-butyl 4-(2-methoxy-2-oxoethyl)piperidine-1-carboxylate:
Figure imgf000558_0001
At O°C, TMSCHN2 (2.0 M in ether, 15 mL) was added dropwise to a mixture of 2-(l-(tert- butoxycarbonyl)piperidin-4-yl)acetic acid (6272 mg, 25.78 mmoL) in Toluene (60 mL) and MeOH (60 mL). After stirring at room temperature for Ih, the mixture was concentrated. The crude tert-butyl 4-(2-methoxy-2-oxoethyl)piperidine-1-carboxylate was used as it is without further purification.
tert-butyl 4-(f7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolof1 ,5-aipyrimidin-5- yl)methyl)piperidine-1-carboxvlate:
Figure imgf000558_0002
tert-butyl 4-(3-cyano-2-oxopropyl)piperidine-1-carboxylate
At -78°C, CH3CN in THF (1505 uL, 28.82 mmoL) was added dropwise to BuLi (2.5 M in hexane, 11.52 mL) in THF (40 mL). After stirring at -78°C for Ih, tert-butyl 4-(2-methoxy-2- oxoethyl)piperidine-1-carboxylate (3708 mg, 14.41 mmoL) in THF (10 mL) was added dropwise in 5 min. The mixture was stirred at -78°C for Ih and -45°C for Ih. At O°C, IN HCl was added carefully to adjust the pH to about 7. The mixture was then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation afforded tert-butyl 4-(3-cyano-2-oxopropyl) piperidine-1 -carboxylate.
tert-butyl 4-((7-aminopyrazolon .5-aipyrimidin-5-yl)methyhpiperidine-1-carboxylate: The crude tert-butyl 4-(3-cyano-2-oxopropyl)piperidine-1-carboxylate was then heated at 100°C overnight with 1 H-pyrazol-5-amine (1197 mg, 14.41 mmoL) in HOAc (25 mL). Concentration provided crude tert-butyl 4-((7-aminopyrazolo[l,5-a]pyrimidin-5- yl)methyl)piperidine- 1 -carboxylate.
tert-butyl 4-((7-(bis((2-(trimethyisilyl)ethoxy)methyπamino)pyrazo[ori ,5-alpyrimidin-5- yl)methyl)piperidine-1 -carboxylate:
The amino analog was treated with SEMCl (72.05 mmoL, 12.71 mL) and DEEA (144.4 mmoL, 25.05 mL) in DCE (100 mL) at 5O°C for 1 h. The mixture was diluted with H2O and then extracted with CH2Cl2 (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation and purification by column chromatography afforded tert-butyl 4- ((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolo[1,5-a]pyrimidin-5- yl)methyl)piperidine-1-carboxylate: LCMS tβ = 3.05 Min (5 min run, UV 254nm)- Mass calculated for, M+ 591.3, observed LC/MS m/z 592.3 (M+H).
Synthesis of tert-butyl 4-((7-(bis(f2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- phenylpyridin-3-yl)pyrazolori ,5-aipyrimidin-5-yl)methyhpiperidine-1-carboxvlate:
Figure imgf000559_0001
tert-butyl 4-((7-(bis((2-(trimethyisilyl)ethoxy)methyl)amino)-3-iodopyrazoio[1,5- a1pyrimidin-5-yl)methyl)piperidine-1-carboxylate:
NIS (288 mg, 1.28 mmoL) was added to a solution of tert-butyl 4-((7-(bis((2- (trimethyIsilyl)ethoxy)methyl)amino)pyrazolo[1,5-a]pyrimidin-5-yI)methyl)piperidine-l - carboxylate (741.6 mg, 1.25 mmoL) in CH3CN (10 mL). The mixture was stirred at room temperature for Ih. Purification by column chromatography afforded tert-butyl 4-((7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-iodopyrazolo[1,5-a]pyrimidin-5-yl)methyl)piperidine- 1-carboxylate: LCMS tR = 3.02 Min (5 min run, UV 254nm). Mass calculated for, M+ 717.2, observed LC/MS m/z 718.3 (M+H).
tert-butyl 4-((7-(bisff2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3- yl)pyrazoiof^5-aipyrimidin-5-yl)methyl)piperidine-i -carboxylate:
2-phenyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (1.32 mmol, 370 mg), K3PO4 (3.04 mmol, 644 mg), and PdCl2(dppf) • CH2Cl2 (0.10 mmol, 83 mg) were added to a solution of tert-butyl 4-((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-todopyrazolo[1,5- a]pyrimidin-5-yl)methyl)piperidine- 1-carboxylate (1.02 mmol, 726 mg) in dioxane (6 mL) and H2O (ImL). The resulting solution was stirred at 100° C under argon for 18 hours. The mixture was diluted with H2O and then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation and purification by column chromatography afforded tert-butyl 4-((7-(bis((2-
(trimethylsslyl)ethoxy)memyl)ammo)-3-(6-phenylpyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5- yl)methyl)piperidine- 1-carboxylate: LCMS IR = 3.47 Min (5 min run, UV 254™)- Mass calculated for, M+ 744.2, observed LC/MS m/z 745.2 (M+H).
(R)- 1 -(4-((7-amino-6-bromo-3-( 6-phenylpyridin-3-yl) pyrazoioM ,5-alpyrimidin-5- yl)methyl)piperidin-1 -vO-2-hydroxypropan-1 -one:
Figure imgf000560_0001
By essentially the same procedure given in Preparative Example above, (R)-1-(4-((7-amino-6- bromo-3-(6-phenylpyridin-3-yl) pyrazolo[l,5-a]pyrimidin-5-yl)methyl)piperidin-1-yl)-2- hydroxypropan-ϊ-one can be prepared. LCMS tβ = 3.58 Min (10 min run, UV 254nm)- Mass calculated for, M-s- 534.13, observed LC/MS m/z 535.55 (M+H). (R)-1-(4-((6-acetyl-7-amJno-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-aipyrimidin-5- yl)methyl)piperidin-1 -yl)-2-hydroxypropan-1 -one:
Figure imgf000561_0001
By essentially the same procedure given in Preparative Example above, (R)-1-(4-((6-acetyl-7- amino-3-(6-pheny[pyriciin-3-yl)pyrazolo[l ,5-a]pyrimidin-5-yl)methyl)piperidin-1-yl)-2- hydroxypropan-1-one was synthesized in a manner similar: LCMS IR = 3.42 Min (10 min run, UV 254nm). Mass calculated for, M+ 498.23, observed LC/MS m/z 499.64 (M+H).
tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3- yl)pyrazolo[1 ,5-aipyrimidin-5-yl)piperazine-1 -carboxviate:
Figure imgf000561_0002
A mixture of 5-chloro-3-(6-phenylpyridin-3-yI)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoio[i,5-a]pyrimidin-7-amine (200 mg, 0.34 mmoL), tert- butyl piperazine-1-carboxylate (224 mg, 1.20 mmoL) and NaHCO3 (116 mg, 1.38 mmoL) in NMP (5 mL) was heated at 13O°C overnight. The mixture was diluted with H2O and then extracted with ethyl acetate (x2). The combined organic layers were washed with brine and dried with Na2SO4. Evaporation and purification by column chromatography afforded tert- butyl 4-(7-(bis((2-(trimethylsiIyl)ethoxy)methyl)amino)-3-(6-phenylpyridin-3-yi)pyrazoio[1,5- a]pyrimidin-5-yl)piperazine-1-carboxylate: LCMS tR = 3.04 Min (5 min run, UV 2S4nm). Mass calculated for, M+ 731.4, observed LC/MS m/z 732.4 (M+H).
(R)-1-(4-(6-acety[-7-amino-3-(6-phenylpyridin-3-yl)pyrazolo[1 ,5-alpyrimidin-5- yl)piperazin-1 -yl)-2-hydroxypropan-1 -one:
Figure imgf000562_0001
By essentially the same procedure given in Preparative Example above scheme, (R)- 1 -(4-(6- acetyl-V-amino-3-tό-phenylpyridin-3-ylJpyrazolotl.5-alpyrimidin-5-yl)piperazin- l-yl)-Z- hydroxypropan-1-one can be prepared in 96% yield, LCMS IR = 2.79 Min (10 min run, UV 254nm). Mass calculated for, M+ 485.2, observed LC/MS m/z 486.3 (M+H).
i -(y-amino-3-(e-phenvIpyridin-3-yl)-5-d -(pyridin-3-ylmethyDpiperidin^- yl)pyrazolon ,5-aipyrimidin-6-yl)ethanone:
Figure imgf000562_0002
A mixture of l-(7-amino-3-(6-phenyipyridin-3-yl)-5-(piperidin-4-yl)pyrazoIo[l ,5-a]pyrimidin- 6-yI)ethanone (0.049 mmol, 20 mg), 3-(bromomethyl)pyridine (0.049 mmol, 12 mg) and DDEA (0.29 mmol, 37 mg) in THF (2mL) was stirred at room temperature overnight. Concentration and purification by prep-LC afforded l-(7-aniino-3-{6-phenylpyridin-3-yl)-5-(l-(pyridin-3- ylmethyl)piρeπdin-4-yl)pyrazoIo[1,5-a]pyrimidin-6-yl)ethanone: LCMS tn = 2.71 Min (10 min run, UV 254nm)- Mass calculated for, M+ 503.24, observed LC/MS m/z 503.99 (M+H).
1-(7-amino-5-(1 -(morpholinosulfonyl)piperidin-4-yl)-3-(6-phenylpyridin-3- yl)pyrazolo[1 ,5a1pyrimidin-6-yl)ethanone :
Figure imgf000563_0001
DIEA (121 mg, 0.94 mmoL) was added to at O°C a mixture of l-(7-amino-3-(6-phenylpyridin- 3-yl)-5-(piperidin-4-yl) pyrazolo [1,5-a]pyrimidin-6-yl)ethanone (64.3 mg, 0.15 mmoL) and morpholine-4-sulfonyl chloride (28.9 mg, 0.15 mmol) in DMF (3 mL). After stirring at O°C for 0.5h and then room temperature for 2 h, the mixture was purified by prep-LC to afford l-(7- amino»5-(l-(morpholinosulfonyl)piperidin-4-yI)-3-(6-phenylpyridin-3-yl)pyrazolo[1,5-
10 a]pyrimidin-6-yl)ethanone, LCMS tR = 3.83 Min (5 min run, UV 2S4nm)- Mass calculated for, M+ 561.66, observed LC/MS m/z 567.66 (M+H).
Synthesis of tert-butyl 4-(7-amino-6-bromo-3-(Quinoiin-3-yl)pyrazolori ,5-a1pyrimidin-5- vQpiperazine-1 -carboxylate
\ 5
Figure imgf000563_0002
To a 20 mL scintillation via! was charged 5-chloro-3-(quinolin-3-yl)-N,N-bis((2- {trimethylsilyl)-ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidtn-7-amine (0.09 mmoi, 50 mg), NMP (1 mL), N,N-diisopropylethylamine (0.36 mmol, 63 μL), and 4-N-Boc-piperazine 0 (0.27 mmol, 50 mg). The resulting solution was heated to 200° C in microwave synthesizer for 30 minutes. Upon completion, the NMP was removed in vacuo as an azeotrope using chlorobenzene. This residue was dissolved in acetonitrile (1 mL) to which N-bromosuccinimide (0.10 mmol, 17.7 mg) was added. This reaction mixture was allowed to stir at room temperature for 2 hours. The reaction mixture was concentrated in vacuo and tert-butyl 4-(7-amino-6-bromo-3-(quinoiin-3-yl)pyrazoio[1 ,5- a]pyrimidin-5-yl)piperazine-1-carboxylate used without further purification.
Synthesis of 6-ch[oro-5-(piperazin-1-yl)-3-(Quinolin-3-vθpyrazoiori ,5-aipyrimidin-7- amine
Figure imgf000564_0001
tert-Butyl 4-<7-amino-6-bromo-3-(quinolin-3-yi)pyrazolo[1 ,5-a]pyrimidin-5- yl)piperazine-1 -carboxylate (0.09 mmol) was charged to a 20 imL scintillation viai. To this vial was added 4N HCI in 1 ,4-dioxane. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo and 6-chloro- 5-(piperazin-1 -yl)-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine was used without further purification.
Synthesis of (4-(7-amino-6-chloro-3-(quinolin-3-yl)pyrazolof1 ,5-alpyrimidin-5- yl)piperazin-1-yl)(thiophen-2-yl)methanone
Figure imgf000564_0002
(4-(7-Amino-6-chloro-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)piperazin-1 - y!)(thiophen-2-yl)methanone was synthesized in a manner similar to the synthesis of (4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)-5,6-dihydropyridin- 1 (2H)-yl)(thiophen-2-yl)methanone, but with 6-chIoro-5-(piperazin-1-yl)-3-(quinoItn-3- yl)pyrazolo[1 ,5-a3pyrimidin-7-amine substituted for 6-bromo-3-(quinolin-3-yl)-5- (1 ,2,3f6-tetrahydropyridin-4-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine. The reaction was purified via reverse-phase preparatory HPLC to yield (4-(7-amino-6-chloro-3-(quinolin- 3-yi)pyrazoio[1 ,5-a]pyrimidin-5-yl)piperazin-1-yl)(thiophen-2-yl)methanone as yellow solid. (M+H = 490.19, retention time = 3.75 min).
Figure imgf000565_0001
Scheme-42
[4-(7-Amino-3-auinonn-3-yl-pyrazolori ,5-a1pyrimidin-5-yl)-phenyll-acetic acid
Figure imgf000565_0002
[4-(4,4,5,5-Tetramethyl-[1 ,3,2]dioxaborolan-2-yl)-phenyl]-acetic acid methyl ester (0.576 mmoi, 160 mg), K3PO4 (0.864 mmol, 183 mg), and PdCI2(dppf) • CH2CI2 (0.029 mmol, 24 mg) was added to a solution of (5-Chloro-3-quinolin-3-yl-pyrazolo[1 ,5- a]pyrimidin-7-yl)-bis-(2-trimethylsilanyl-ethoxymethy[)-amine (0.288 mmol, 160 mg) in dioxane (2 ml_). To this suspension was added distilled H2O (0.2 ml_). The resulting solution was stirred at 100° C under an argon atmosphere for 18 hours. The reaction mixture was concentrated in vacuo. The crude mixture was dissolved in 2:1 MeOH:H2O (1.5 ml_) and was treated with 2N NaOH(aq) (1.0 ml_). The resulting solution was stirred at room temperature for 18 hours. Aqueous hydrochloride solution (1.0 N, 4 ml) was added to the reaction mixture and the resulting solution was stirred at 65 °C for 2 h. The reaction mixture was concentrated and purified by prep- LC to afford the title compound (70 mg, 62% yield): LC/MS RT = 3.21 min. Mass calculated for, M+H 396.14, observed 396.14.
Figure imgf000566_0001
r4-(7-Amino-6-bromo-3-αuinolin-3-yl-pyrazoio[1 ,5-a]pyrimidin-5-yl)-phenyl)-acetic acid A solution of NBS (32 mg, 0.179 mmol) in CH3CN (1 mi_) was added to a mixture of [4-(7-amino-3-quinoiin-3-yl-pyrazolo[1 ,5-a]pyrimidin-5-yl)-phenyl]-acetic acid {70 mg, 0.179 mmoi) in CH3CN (3 ml_). After stirring at room temperature for 1 h, the solvent was evaporated in vacuo and the residue was purified by prep-LC to afford titled compound (22.9 mg, 27%). LC/MS RT = 3.44 min. Mass calculated for, M+H 474.05, observed 474.05.
By essentially the same procedure given in Scheme 42, the compounds listed in Table 11 can be prepared.
Table 11
Figure imgf000566_0002
Figure imgf000567_0001
Figure imgf000568_0001
Synthesis of tert-butyl 4-(7-(bis(f2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolof1 ,5-aipyrimidin-5-yl)-5,6-dihydropyridine-1 (2H)-carboxylate
Figure imgf000569_0001
To a 40 mL scintillation vial was charged 5-chloro-3-(quinoiin-3-yl)-N,N-bis((2- (trimethylsiiyl)-ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine (0.86 mmol, 480 mg), tert-butyl 4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-5,6-dihydropyrid!ne-1 (2H)- carboxylate (1.30 mmol, 400 mg), K3PO4 (2.60 mmol, 550 mg), and PdCI2(UpPf)-CH2CI2 (0.086 mmol, 70 mg). To this reacton mixture was added a 9:1 solution of 1 ,4-dioxane:H2O (10 mL). The vial was flushed with argon, sealed with Teflon tape, and stirred at 100° C 18 hours. After 18 hours, the reaction was concentrated in vacuo and the residue purified via silica gel chromatography to yield tert-butyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyhmidin-5-yl)-5,6-dihydropyridine-1 (2H)-carboxylate (586 mg, 833 mmol, 97% yield) as yellow solid.
.Synthesis of 3-(αuinolin-3-yl)-5-(1 ,2.3.6-tetrahydropyridin-4-yl)pyrazoioM .5- aipyrimidin-7-amine
Figure imgf000569_0002
tø/t-Butyl 4-(7-(bis((2-(trimethylsiiyl)ethoxy)methyl)amino)-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)-5,6-dihydropyridine-1(2H)-carboxylate (0.057 mmol, 40 mg) was charged to a 20 mL scintillation vial containing 1 mL TFA. The reaction mixture was stirred at room temperature. The reaction mixture was concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield 3-(quinolin-3-yl)-5- (1 ,2,3,6-tetrahydropyridin-4-y!)pyrazolo[1 ,5-a]pyrimidin-7-amine (m+H = 343.31 , retention time = 2.05 min) Synthesis of 6-bromo-3-(ouinolin-3-yl)-5-( 1 ,2,3,6-tetrahydropyridin-4-yl)pyrazolori .5- aipvrimidin-7-amine
Figure imgf000570_0001
tø/t-Butyl 4-(7-(bis((2-{trimethy!silyl)ethoxy)methyI)amino)-3-(quinolin-3- yl)pyrazoio[1 ,5-a]pyrimidin-5-yl)-5,6-dihydropyridine-1 (2H)-carboxyiate (0.071 mmol, 50 mg) was charged to a 20 ml_ scintillation vial containing 1 mi_ acetonitrile. To this solution was added N-bromosuccinimide (0.078 mmoi, 14 mg). The reaction mixture was allowed to stir at room temperature for 1 hour. After 1 hour, the reaction was concentrated in vacuo and the crude intermediate was dissolved in 1 mL TFA. The reaction mixture was stirred for 1 hour at room temperature and concentrated in vacuo to yield 6-bromo-3-(quinolin-3-y!)-5-(1 ,2,3,6-tetrahydropyridin-4-yl)pyrazolo[1 ,5- a]pyrimidin-7-amine as yellow solid.
Synthesis of 6-chloro-3-(quinolin-3-yl)-5-(1 ,2,3,6-tetrahydropyridin-4-yl)pyrazolof 1 ,5- aipyrimidin-7-amine
Figure imgf000570_0002
6-Chloro-3-(quinolin-3-yl)-5-(1 ,2,3,6-tetrahydropyridin-4-yl)pyrazoio[1 ,5-a3pyrimidin-7- amine was synthesized in a manner similar to the synthesis of 6-bromo-3-(quinolin-3- yl)-5-(1 ,2,3,6-tetrahydropyridin-4-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine, but with N- chlorosuccinimide substituted for N-bromosuccinimide. Synthesis of 2-(4-(7-amino-6-bromo-3-(αuinolin-3-yl)pyrazolof1 ,5-alpyrimidin-5-yl)-5,6- dihydropyridin-1 (2H)-yl)acetic acid
Figure imgf000571_0001
6-Bromo-3-{quinolin-3-yl)-5-{1 ,2,3,6-tetrahydropyrid!n-4-yl)pyrazo[o[1 ,5-a]pyrimidin-7- amine (0.035 mmol) was charged to a 20 mL scintillation vial. Then DMF (1 ml), N1N- diisopropylethylamine {0.14 mmo!, 25 μl_), and tert-butyl 2-bromoacetate (0.035 mmol, 5.2 μl_) were added, respectively. The resulting reaction mixture was stirred at 50Q C 18 hours. After 18 hours, the reaction was concentrated in vacuo and treated with 1 mL TFA. This reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield 2-(4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)-5,6-dihydropyridin-1 (2H)-yl)acetic acid as yellow solid. (m+H = 479.17, retention time = 2.09 min)
Synthesis of (4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazoiori ,5-a1pyrimidin-5-yl)-5.6- dihydropyridin-1 (2H)-yl)(thiophen-2-yl)methanone and (4-(7-amino-3-(quinolin-3- yl)pyrazolo[1 ,5-alpyrimidin-5-yl)-5,6-dihydropyridin-1(2H)-yl)(thiophen-2-yl)methanone
Figure imgf000571_0002
SCHEME-43
6-Bromo-3~(quino!in-3-yl)-5-(1 ,2,3,6-tetrahydropyridin-4-yl)pyrazolo[1 ,5-a]pyrimidin-7- amine (0.035 mmol) was charged to a 20 mL scintillation vial. Then DMF (1 ml), N1N- diisopropylethylamine (0.14 mmol, 25 μL), thiophene-2-carboxylic acid (0.039 mmol, 5 mg), and EDC (0.039 mmol, 7.4 mg) were added, respectively. The resulting reaction mixture was stirred at room temperature for 18 hours. Upon completion, the reaction mixture was concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield (4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yI)-5,6- dihydropyridin-1(2H)-yl)(thiophen-2-yl)methanone (m+H = 531.06, retention time = 3.74 min). Also recovered was the dehalogenated product, (4-(7-amino-3-(quinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl}-5,6-dihydropyridin-1 (2H)-yl)(thiophen-2-y])methanone (m+H = 453.24, retention time = 3.51 min).
Synthesis of (4-(7-amino-6-chloro-3-(quino[in-3-yl)pyrazolof1.5-alpyrimidin-5-yl)-5,6- dihydropyridin-1(2H)-yl)(thiophen-2-yl)methanone
Figure imgf000572_0001
(4-(7-amino-6-chloro-3-(quinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)-5I6-dihydropyridin- 1 (2H)-yl)(thiophen-2-yl)methanone was synthesized in a manner similar to the synthesis of (4-(7-amino-6-bromo-3-(quinolin-3-yl)pyrazolo[1 ,5~a]pyrimidin-5-yl)-5,6- dihydropyridin-1 (2H)-yl)(thiophen-2-yi)methanone, but with 6-chloro-3-(quinolin-3-yl)- 5-(1 ,2,3,6-tetrahydropyridin-4-yI)pyrazolo[1 ,5<|pyrimidin-7-amine substituted for 6- bromo-3-(quinolin-3-yl)-5-(1 ,2,3,6-tetrahydropyridin-4-yl)pyrazolo[1 ,5-a]pyrimidin-7- amine. The reaction mixture was purified via reverse-phase preparatory HPLC to yield (4-(7-amino-6-chloro-3-(quino!in-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yi)-5,6- dihydropyridin-1(2H)-yl)(thiophen-2-yl)methanone as yellow solid. (m+H = 487.17, retention time = 3.73 min)
Synthesis of 4-bromo-1 -((2-(trimethylsilyl)ethoxy)nπethyπ-1 H-pyrazole SEM-Cl
Figure imgf000573_0001
To a 100 mL roundbottom flask was charged 4-bromopyrazole (13.6 mmol, 2.00 g), K2CO3 (20.4 mmol, 2.82 g), and DMF (20 mL). To this suspension was added 2- (trimethylsilyl)ethoxymethyl chloride (15.0 mmol, 2.64 mL). This suspension was allowed to stir at room temperature 18 hours. The reaction mixture was then concentrated in vacuo, dissolved in 20 mL DCM, and filtered through celite. The crude mixture was purified via silica gel chromatography (0% to 80% ethyl acetate in hexanes gradient) to yield 4-bromo-1 -({2-(trimethylsilyI) ethoxy) methyl)-1 H-pyrazole (2.53 g, 9.12 mmol, 67% yield) as clear oil.
Synthesis of 4-(1 ,4-dioxaspiro[4.51dec-7-en-8-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)- 1 H-pyrazole
Figure imgf000573_0002
To a 10-20 mL microwave vessel was charged 4-bromo-1 -({2- (trimethylsilyl)ethoxy)methyl)-1 H-pyrazole (0.60 mmol, 166 mg), 4,4,5,5-tetramethyl-2- (1 ,4-dioxaspiro[4.5]dec-7-en-8-yl)-1 ,3,2-dioxaborolane (0.84 mmol, 224 mg), K3PO4 (1.80 mmol, 382 mg), PdCI2{dppf) ■ CH2CI2 (0.06 mmoi, 50 mg) and 9:1 Dioxane:H2O (15 mL). The suspension was sonicated to break up salts, flushed with argon, sealed, and heated to 160° C for 45 minutes in microwave synthesizer. The reaction was concentrated in vacuo, and the residue purified via silica gel chromatography (0% to 60% ethyl acetate in hexanes) to yield 4-(1 ,4-dioxaspiro[4.5]dec-7-en-8-yl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 H-pyrazole (115 mg, 0.34 mmol, 57% yield) as clear oil. Synthesis of 4-(1 ,4-dioxaspiro|"4.5ldecan-8-y1H -((2-(trimethyisilyl)ethoxy)methyl)-1 H- pyrazoie
EtOAc
Figure imgf000574_0001
4-{1 ,4-Dioxasptro[4.5]dec-7-en-8-yl)-1 -((2-(trimethylsiiyl)ethoxy)methyl)-1 H-pyrazole (505 μmol, 170 mg) was dissolved in 10 ml_ EtOAc in a 50 ml_ roundbottom flask. The flask was flushed with argon and 5% palladium on carbon (50 mg) was added. The flask was sealed, degassed under vacuum, and hydrogen gas was added with a balloon. The reaction mixture was stirred at room temperature under hydrogen atmosphere for 18 hours. The reaction was then filtered through ceiite to yield 4-(1 ,A- dioxaspiro[4.5]decan-8-yl)-1 -((2-(trimethylsilyl)ethoxy)methyl)-1 H-pyrazole (165 mg, 487 μmol, 97% yield) as a clear oil.
Synthesis of 4-(1-((2-(trimethylsilyl)ethoxy)methy1H H-pyrazol-4-yl)cvclohexanone
EtOH
Figure imgf000574_0002
4-(1 ,4-dioxaspiro[4.53decan-8-yi)-1-((2-(trimethylsilyI)ethoxy)methyl)-1 H-pyrazoie (487 μmol, 165 mg) was dissolved in EtOH (10 m L) in a 20 ml_ scintillation vial. To this solution was added H2O (3 ml_) followed by 4N HCI in 1 ,4-dioxane. The resulting reaction mixture was stirred at room temperature for 60 hours. At this point, the reaction was quenched with saturated
Figure imgf000574_0003
and extracted with DCM (x3). The combined organics were dried over Na2SO4 to yield 4-(1-((2- (trimethylsilyl)ethoxy)methyl)-1 H-pyrazol-4-yl)cyclohexanone (141 mg, 480 μmol, 98% yield) as white solid.
Synthesis of 4-(1 -((2-(trimethylsiiyl)ethoxy)methyl)-1 H-pyrazol-4-yl)cvclohex-i -enyl trifluoromethanesulfonate
Figure imgf000575_0001
4-(1 -((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)cyclohexanone (480 μmol, 141 mg) was added to a 50 mi_ roundbottom flask containing anhydrous THF (5 ml_). The flask was flushed with argon, sealed, and cooled to -78° C in dry ice/isopropanol bath. To this solution was added dropwtse LDA (1.5 M is cyclohexane, 0.72 mmol, 480 μL). This solution was stirred at -78° C for 30 minutes. To this solution was added 2-[N1N- bis(trifluoromethylsulfonyl)amino]-5-chloropyridine (0.72 mmol, 282 mg). The resulting reaction mixture was allowed to gradually warm to room temperature as it stirs for 18 hours. After 18 hours, the reaction mixture was concentrated in vacuo and purified via silica gel chromatography to yield 4-(1-((2-(tπmethylsilyl)ethoxy)methyl)-1H-pyrazol-4- yl)cyclohex-1-enyl trifluoromethanesulfonate (115 mg, 270 μmol, 56% yield) as clear oil.
Synthesis of 4-(4-(4,4,5,5-tetramethyl-1.3.2-dioxaborolan-2-vncvclohex-3-enyl)-1 -((2- (trimethylsiiyl)ethoxy)methyl)-1 H-pyrazole
Figure imgf000575_0002
To a 20 mL scintillation vial was charged 4-(1 -((2-(trtmethylsilyl)ethoxy)methyl)-1 H- pyrazol-4-yl)cyclohex-1 -enyl trifluoromethanesulfonate (0.047 mmol, 20 mg), bis(pinacolato)diboron (0.070 mmol, 18 mg), potassium acetate (0.094 mmol, 9.2 mg), PdCI2(PPh3)2 (0.0014 mmol, 1 mg), triphenylphosphine (0.0028 mmol, 0.8 mg), KBr (0.07 mmol, 8.4 mg), and toluene (0.5 mL). The vial was flushed with argon and sealed. The reaction was then stirred at 60° C for 18 hours. After 18 hours, the reaction was filtered through celite, and 4-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaboroJan- 2-yl)cyclohex-3-enyl)-1 -((2-(trimethylsilyi)ethoxy)methyl)-1 H-pyrazole was used without further purification. Synthesis of 5-(4-(1 H-pyrazoi-4-yl)cvciohex-1-enyl)-3-(αuinolin-3-yl)pyrazoio[1 ,5- aipyrimidin-7-amine
TFA
Figure imgf000576_0001
To a 2 ml microwave vessel was charged 5-chloro-3-(quinolin-3-yl)-N,N-bis({2- (trimethylsilyl)ethoxy)methy])pyrazolo[1 ,5-a]pyrimidin-7-amine (0.043 mmol, 23.5 mg), 4-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)-1 -((2- (trimethylsilyi)ethoxy)methyl)-1 H-pyrazole (0.047 mmol), K3PO4 (130 mmol, 27 mg), and PdCi≥(dppf) • CH2CI2 (0.0043 mmoi, 3.5 mg). To this mixuture was added 9:1 dioxane: H2O (1 mL). The vessel was flushed with argon and heated to 160° C for 30 minutes in microwave synthesizer. Upon completion, the reaction was concentrated in vacuo and the residue was dissolved in 1 mL TFA. This reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo and purified via reverse-phase preparatory HPLC to yield 5-(4-(1 H-pyrazo!-4-yl) cyclohex~1-enyl)-3~(quinolin-3-yl) pyrazolo [1 , 5-a] pyrimidin-7-amine as yellow solid. (m+H = 408.30, retention time = 3.16 min)
Synthesis of 5-(4-(1 H-pyrazoi-4-vOcvclohex-i -enyl)-6-bromo-3-(αuinolin-3- yl)pyrazolof1.5-aipyrimidin-7-amine
Figure imgf000576_0002
5-(4-(1 H-pyrazoi-4-yl)cyclohex-1 -enyl)-6-bromo-3-(quinolin-3-yl)pyrazo!o[1 ,5- ajpyrimidin-7-amine was synthesized in a manner similar to the synthesis of 6-bromo- 3-(quinoiin-3~yl)-5-(1 ,2,3,6-tetrahydropyridin-4-yl)pyrazolo[1 t5-a]pyrimidin-7-amine, but with 6-bromo-3-{quinolin-3-yl)-N,N-bis((2-(trimethylsilyi)ethoxy)methyl)-5-(4-(1-((2- (trimethylsilyl)ethoxy)methyl)-1 H-pyrazo!-4-yl)cyclohex-1 -enyl)pyrazolo[1 ,5- a]pyrimidin-7-anπine substituted for tert-Butyl 4-{7-{bis((2-
(trimethylsilyl)ethoxy)methyl)amino)-3-(quinolin-3-yl)pyrazoio[1 ,5--a]pyπmicIin-5-yl)-5,6- dihydropyridine-1 (2H)-carboxylate. The reaction mixture was purified via reverse- phase preparatory HPLC to yield 5-(4-(1 H-pyrazoi-4-yi) cyclohex-1-enyl)-3-(quinolin-3- yi) pyrazolo [1 ,5-a]pyrimidin-7-amine as yellow solid. (M+H = 486.12, retention time = 3.43 min.
By essentially the same procedure given in Scheme 43, the compounds listed in Table 12 can be prepared.
Table 12
Figure imgf000577_0001
Figure imgf000578_0001
Figure imgf000579_0001
Scheme 44
Synthesis of 3-(quinolin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)-5- vJnylpyrazolof1 ,5-alpyrinnidin-7-amine
Figure imgf000579_0002
To 5-chloro-3-tquinolin-3-yl)-N.N-bisftZ-^rimethylsilyl)ethoxyJmethyl)pyrazoloCi .5- a]pyrimidin-7-amine (0.9 g, 1.64 mmol) in a 20 ml_ vials was added 1 ,4-dioxane (10 ml_), tributyl(vinyl)tin (0.5 mL, 1.71 mmol) and Pd(PPh3J4 (0.1 g, 0.08 mmol). The vial was flushed with argon, sealed and heated at 100 °C for 16 hours, at which time LC/MS analysis confirmed full conversion of the starting material to the product. The reaction mixture was filtered through a mixture of silica (8 g) and KF (2 g), and EtOAc was used to wash the filter pad. Concentration of the filtrate gave a brown oil (1.4 g). Further purification via gradient column chromatography on silica eltuting with 5% to 40% EtOAc/hexanes gave 3-(quinoiin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)-5- viny]pyrazolo[1 ,5-a]pyrimidin-7-amine as a yellow oil (0.7 g, 1.3 mmol, 78%). LCMS: 2.55 mins, m/z = 548.2 (MH+). Synthesis of 7-(bis((2-(tnmethylsilyl)ethoxy)methyl)amino)-3-(quinoiin-3- yl)pyrazoloH ,5-aipyrimidine-5-carbaidehyde
Figure imgf000580_0001
To 3-{quinolin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)-5-vinylpyrazolo[1 ,5- a]pyrimidin-7-amine (0.7 g, 1.3 mmol) in 1 ,4-doixane (10 ml_) was added 2.5 wt% OsO4 in 1 ,4-dioxane (1.0 g, 0.02 mmoi), 2,6-lutidine (0.6 ml_, 5.2 mmol) and H2O (2 mL), and the r esulting mixture was stirred at room temperature for 20 minutes. NaIO4 (0.8 g, 3.81 mmol) was then added and the reaction mixture was stirred at room temperature for 16 hours. LC/MS analysis at that time showed the diol intermediate still present. Therefore, more NaIO4 (0.3 g, 1 ,3 mmol) was added, and the reaction mixture was stirred for 12 hours for fuil conversion to the product. Saturated Na2S2Oa solution (10 mL) was added and the mixture was stirred for 10 minutes. Organics were then extracted with CH2CI2 (4 x 40 mL), dried (Na2SO4) and concentrated in vacuo to give 7-(bis((2-(trimethy!silyl)ethoxy)methyl)amino)-3-(quinolin-3-yi)pyrazolo[1 ,5- a]pyrimidine-5-carbaldehyde as a yellow solid (0.6 g, 1.2 mmol, 92%). LCMS: 2.67 mins, m/z = 550.0 (MH+).
Synthesis of 5-(morpholinomethyl)-3-(Quinolin-3-vS)-N, N-bis((2-(trimethylsilvh- ethoxy)methyl)pyrazolo[1 ,5-aipyrimidin-7-amine
Figure imgf000580_0002
To 7-(bis((2-(trimethyIsilyl)ethoxy)methyl)amino)-3-(quinolin-3-yi)pyrazolo[1 ,5- a]pyrimidine-5-carbaldehyde (0.15 g, 0.3 mmol) in EtOH (10 mL) was added morphoiine {0.12 mL, 1.4 mmol) and AcOH (0.2 mL), and the resulting mixture was stirred at room temperature for 15 minutes. NaBH3CN (0.12 g, 2.0 mmol) in EtOH (1.5 mL total) was then added and the reaction mixture was stirred at room temperature for 16 hours, at which time LC/MS analysis confirmed full consumption of starting material. Saturated NaHCO3 solution (~ 20 mL) was added and the mixture was stirred for 30 minutes. The mixture was transferred to a separatory funnei with CH2CI2 (30 mL) and H2O (10 mL) and brine (20 mL) was then added. Organics were extracted with CH2CI2 (4 x 30 mL), dried (Na2SO4) and concentrated in vacuo to give crude 5- (morpholinomethyl)-3-(quinolin-3-yl)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine as a brown oil (0.2 g). LCMS: 2.01 mins, m/z = 621.3 (MH+).
Synthesis of 5-(morphoJinomethyD-3-(αuinolin-3-vS)pyrazolori ,5-alpyrimidin-7-amine
Figure imgf000581_0001
To crude 5-(morpholinomethyl)-3-(quinoiin-3-yl)-N,N-bis((2-(trimethylsilyl)- ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin~7-amine (0.2 g) in a 20 mL vial was added EtOH (4 mL) and then 4M HCI/dioxane (1.5 mL, 6.0 mmol). The vial was capped and heated at 85 °C for 16 hours. On cooling, saturated NaHCO3 solution (~5 mL) was added, and the reaction mixture was stirred for 30 minutes. The reaction mixture was then transferred to a separatory funnel using CH2Cl2 (20 mL) and H2O (20 mL) and saturated NaHCO3 solution (20 mL) and brine (20 mL) were added. Organics were extracted with CH2CI2 (4 x 30 mL), dried (Na2SO4) and concentrated in vacuo to give crude 5-(morpholinomethyl)-3-(quinolin-3-yl) pyrazolo [1, 5-a] pyrimidin-7-amine as a brown solid (0.04 g). LCMS: 0.78 mins, m/z = 361.1 (MH+). Synthesis of 6-bromo-5-(morphoiinomethyl)-3-(quinolin-3-yl)pyrazolo[1 ,5-aipyrimidin- 7-amine
Figure imgf000582_0001
To crude 5-(morpholinomethyl)-3-(quinolin-3-yl) pyrazolo[1 ,5-a]pyrimidin-7-amine (44 mg) in CH3CN {8 mL) was added N-bromosuccinimide (22 mg, 0.12 mmol), followed by additional CH3CN (4 mL). The reaction was then stirred at room temperature for 30 minutes, at which time LC/MS analysis confirmed full conversion of the starting material to the product. The reaction mixture was concentrated in vacuo and purified by preparative HPLC to give 6-bromo-5-(morpholinomethyl)-3-(quinolin-3- yi)pyrazolo[1 ,5-a]pyrimidin-7-amine as a yellow solid (8.7 mg, 0.02 mmol, 7% over three steps). LCMS: 0.82 mins, m/z = 439.0 and 441.0 (MH+).
Related reductive amination analogs were prepared as described in Preparative Example included in the following table. In most cases, reduction of the aldehyde to the primary alcohol was a competing reaction to the desired reductive amination. Generally, reactions giving >80% reductive amination product progressed without purification whereas those with >20 % primary alcohol were purified by silica gel column chromatography before continuing to the de-SEM reaction.
By essentially the same procedure given in Scheme 44, the compounds listed in Table 13 can be prepared.
Table 13
Figure imgf000582_0002
Figure imgf000583_0001
Figure imgf000584_0001
Figure imgf000585_0001
Figure imgf000586_0001
Figure imgf000587_0001
Figure imgf000588_0001
Figure imgf000589_0001
Figure imgf000590_0001
Figure imgf000591_0001
Figure imgf000592_0001
Figure imgf000593_0001
Figure imgf000594_0001
Figure imgf000595_0001
Figure imgf000596_0001
Synthesis of 6-bromo-5-ff(2R.6SV2.6-dimethyltetrahydro-2H-Dyran-4- ylamino)methyl)3-(6-fluoroQuinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-7-amine: Part A
Figure imgf000597_0001
To a solution of (2R,6S)-2,6-dimethyldihydro-2H-pyran-4(3H)-one (0.22g, 1 ,7mmol) in MeOH (11.5ml) at room temperature was added ammonium acetate (1.3g, 16.9mmol) and sodium borohydride (0.07g, l.8mmol) slowly and the reaction mixture was stirred for 16 h, at which time LC/MS analysis confirmed full consumption of starting material. The mixture was transferred to a separatory funnel with CH2Cl2 (30ml) and H2O (10ml) and brine (20ml) was then added. Organics were extracted with CH2Cl2 (4 x 30ml), dried (Na2SO4) and concentrated in vacuo to give crude product 2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-amine. Part B:
Figure imgf000597_0002
2 3
To 2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-amine (crude) in CH3CN (3.2ml) was added
N,N-diisopropylethylamine (0.9ml) followed by di-tørf-butyl dicarbonate (1.9g) and the resulting mixture was stirred at room temperature for Ih, at which time LC/MS confirmed full conversion of starting material to product. Solvent was removed in vacuo and the crude was redissoived in DCM (20ml), washed with water (1 x 5ml), brine (1 x 5ml), dried over MgSO4. Solvent was removed in vacuo to give tert-butyl (2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4- ylcarbamate as a crude product which was used for the next step without any purification.
Part C:
Figure imgf000598_0002
To tert-butyl (2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-ylcarbamate (crude, 0.6g) was added 4M HCl in 1,4-dixoane (15ml) at r.t. It was stirred further at room temperature for 2 h, at which time LC/MS analysis confirmed full consumption of starting material. Solvent was removed in vacuo to get the desired (2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-amine as an HCl salt.
Part D:
Figure imgf000598_0001
To crude (2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-amine (O.lδg) in DCE (4.2ml) and Ethanol (4.2ml) was added 7-(bis((2-(trimethylsilyI)ethoxy)methyl)amino)-6-bromo-3-(6- fluoroquinolin-3-yl)pyrazolo[l,5-a]pyrimidine-5-carbaldehyde (0.19g, 0.3mmol) followed by AcOH (0.03ml, O.όmrøol). After stirring for 30 minutes at room temperature, NaBH3CN
(37.8mg, O.όmmol) was added and stirring was continued further for 30 minutes more. LC/MS showed no starting material remaining. After the solvent was rotoevaporated, the crude was dissolved with DCM (20ml), washed with sat. NaHCO3 (1 x 5ml), water (1 x 5ml), brine (1 x 5ml), dried over MgSO4. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with EtOAc/Hexanes (0-60%) gave desired 6- bromo-5-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-ylamino)methyl)-3-(6-fluoroquinolin- 3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methy[)pyrazolo[ 1 ,5-a]pyrimidin-7-amine (68 mg, 30%).
Part E 6-bromo-5-(((2R,6S)-2.6-dimethyltetrahyclro-2H-Dyran-4-ylamino)methyπ-3-(6- fluoroquinolin-3-yl)pyrazoloπ,5-a1pyrimidin-7-amine:
Figure imgf000599_0001
To a 6-bromo-5-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-ylamino)methyl)-3-(6- fluoroquinolin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)niethyl)pyrazolo[1,5-a]pyrimidin-7- amine (69.3mg, 0.09mmol) in TFA (2ml) was added few drops of water and stirring continued for 2h at room temperature. LC/MS showed no starting material remaining. TFA along with water was rotoevaporated. This crude compound was submitted to the analytical group for purification to afford the desired 6-brorno-5-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4- ylamino)methyl)-3-(6-fluoroquinolin-3-yl)pyrazolo[1,5-a]pyrimidin-7-amine. LCMS: 3.157 mins, m/z = 499.0 (MH+).
Synthesis of 6-bromo-3-(6-fluoroquinolin-3-yl)-5-((3-methoxytetrahydro-2H-pyran-4- ylamino)methyθpyrazolori ,5-aipyrimidin-7-amine:
M
Figure imgf000599_0002
SCHEME- 45
Above compound was prepared by essentially the same procedure (1) as to compound 7 is prepared. However exact configuration has not been determined. Isomer 1: LCMS: 1.942 mins, m/z = 501.1 (MH+) Isomer 2: LCMS: 2.706 mins, m/z = 501.0 (MH+)
Synthesis of 6-bromo-3-(6-fluoroquinolin-3-ylV5-((1 -
(methylsulfonylmethyl)cvclopropylamino)methyπpyrazolof1.5-aipyrinnidin-7-amiπe:
Figure imgf000600_0001
SCHEME-46
/nthesis of ethyl 1 -(tert-butoxycarbonylamino)cvclopropanecarboxylate:
To ethyl 1-aminocyclopropane-1-carboxylic acid ethyl ester hydrochloride (7.9g, 48mmol) in DCM (400ml) was added N,N-diisopropylethylamine (21.7ml, 124.7mmol) followed by di- tert-butyl dicarbonate (1 l.5g, 52.8mmoi) at O°C, and the resulting mixture was stirred at room temperature for 16h, at which time LC/MS confirmed full conversion of starting material to product. Reaction mixture was transferred in to the seperatory funnel and washed with water (1 x 75ml), brine (1 x 75ml), dried over MgSO4. Solvent was removed in vacuo to get desired ethyl l-(tert-butoxycarbonylamino)cyclopropanecarboxylate as a crude product which was used for the next step without any purification.
Synthesis of tert-butyl Hhydroxymethyπcvclopropylcarbarnate:
While under argon atmosphere, LiBH4 (0.73g, 33.6mmol) was suspended in Et2O (18ml). To this mixture was added dropwise a solution of ethyl 1 -(tert- butoxycarbonylamino)cyclopropane carboxylate (5g, 21.8mmol) in Et2O (18ml). The reaction mixture was stirred at room temperature for 2 h and then quenched by slow addition of MeOH (18ml), After being stirred overnight, the reaction mixture was poured into an equal volume of sat. NH4Cl (54ml). The Et2O layer was separated, and the aqueous layer was extracted four times with Et2O. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with EtOAc/Hexanes (50-100%) gave desired tert-butyl 1 -(hydroxymethyl)cyclopropyl carbamate as a white solid (3.14g, 77%).
Synthesis of (1 -(tert-butoxycarbonylamino^cvclopropyl)methylmethanesulfonate:
To tert-butyl l-OαydroxymethyOcyclopropylcarbamate (1.Ig, 5.83mmo!) in DCM (47ml) was added N,N-diisopropylethylamine (1.3ml, 7.67mmol) followed by methanesulfonylchloride (0.5ml, 6.4mmol) at 0°C, and the resulting mixture was stirred at room temperature for 16h, at which time LC/MS confirmed full conversion of starting material to product. Reaction mixture was transferred in to the seperatory funnel and washed with sat. NaHCOj (1 x 10ml), water (1 x 10ml), brine (1 x 10ml), dried over MgSO4. Solvent was removed in vacuo to get desired (1- (tert-butoxycarbonylamino)cyclopropyl)methylmethanesulfonate as a crude product ( 1.38g) which was used for the next step without any purification.
Synthesis of tert-butyl i-(methylthiomethyDcvciopropylcarbamate:
A mixture of (l-(tert-butoxycarbonylamino)cyc[opropyl)methy!methanesulfonate (1.38g,
5.2mmol), and NaSCH3 (0.48g, 6.8mmol) in DMF (21ml) was stireed at room temperature for 2 h, at which time LC/MS confirmed full conversion of starting material to product (No Starting material is present). Reaction mixture was diluted with EtOAc (250ml), and. washed with water (3 x 60ml), brine (I x 60ml) and dried over MgSO4. Solvent was removed to give the tert-butyl l-(methylthiomethyl)cyclopropylcarbamate as a crude product which was used for the next step without any purification.
Synthesis of tert-butyl i -(methyisulfonylmethyDcvciopropylcarbamate:
A mixture of tert-butyl l-tmethylthiomethyDcyclopropylcarbamate (crude), Oxone (4.8g, 7.8mmol) and NaHCO3 (2.6g, 3 L3mmol) in MeOH (28ml) and Water (7ml) was stirred at room temperature for 1.5 h, at which time LC/MS analysis confirmed full consumption of starting material. Solvent was removed in vacuo and the crude was redissolved in EtOAc (100ml), washed with water (1 x 20ml), brine (I x 20ml), dried over MgSO4. Solvent was removed in vacuo to give tert-butyl l-(methylsulfonylmethyl) cyclopropy [carbamate as a crude product (white solid, l.lg) which was used for the next step without any purification.
Synthesis of i-(methylsulfonylmethyl)cyclopropanamine:
To tert-butyl l-(methylsulfonylmethyl)cyclopropyicarbamate_(crυde, 1.05g) in DCM (7ml) was added 4M HCl in 1,4-dixoane (16ml) at r.t. It was stirred further at room temperature for 1 h, at which time LC/MS analysis confirmed full consumption of starting material. Solvent was removed in vacuo and the solid was washed with ether to get the desired 1 - (methytsulfonylmethyl)cyclopropanarnine as an HCl salt.
Synthesis of 6-bromo-3-(6-fluoroquinolin-3-yl)-5-(f 1 -(methyisulfonylmethyhcvclo propylamino)methyl)pyrazolori,5-aipyrimidin-7-amine:
6-bromo-3-(6-fluoroquinolin-3-yI)-5-((l-(methylsulfonylmethyl)cyclo propylamino)methyl)pyrazolo[1,5-a]pyrimidin-7-amine was prepared by essentially the same procedure (1) as to compound 7 is prepared. LCMS: 2.52 mins, m/z = 519.0 (MH+).
Synthesis of (1 -((7-(bis(f2-(trinnethylsilyl)ethoxy)methyπamino)-6-bromo-3-(6- fiuoroQuinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)methylamino)cyclopropyl)methanoi:
Figure imgf000603_0001
To l-aminocyclopropyl)methanol (0.22g, 2.5mmol) in DCE (7ml) and Ethanol (7ml) was added -(bis{(2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-fluoroquinolin-3- yl)pyrazoIo[1,5-a]pyrimidine-5-carbaldehyde (0.32g, O.Smmol) followed by AcOH (0.064ml, Immol). After stirring for 30 minutes at room temperature, NaBH3CN (63mg, Immol) was added and stirring was continued further for 30 minutes more. After the solvent was rotoevaporated, the crude was dissolved with DCM (40ml), washed with sat. NaHCO3 (I x 10ml), water (1 x 10ml), brine (1 x 10ml), dried over MgSO^ Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with EtOAc/Hexanes (0-100%) gave slightly impure desired (1 -((7 -(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-fluoroquinolin-3-yOpyrazolo[1,5- a]pyrimidin-5-yl)methylamino)cyclopropyl)methanol (320mg).
Synthesis of tert-butyl (7-(bis((2-(trimethylsilyl)ethoχy) methyl)amino)-6-bromo-3-(6- fluoroquinolin-3-yl)pyrazolori 3-a1pyrimidin-5-vOmethyl(1 - (hydroxymethyDcvciopropyl)carbamate:
Figure imgf000603_0002
To ( 1 -((7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-fluoroquinolin-3- yl)pyrazolo[1,5-a]pyrimidin-5-yl)methylamino)cyclopropyl)methanol (0.17g, 0.24mmol) in dichloroethane (2ml) was added N,N-diisoproρylethyIamine (0.09ml, 0.49mmoi) followed by di-tert-butyl dicarbonate (O.lόg, 0.73mmol) and the resulting mixture was stirred at room temperature for 16 h, at which time LC/MS confirmed full conversion of starting material to product. The mixture was diluted with DCM (20ml), washed with water (1 x 5ml), brine (1 x 5ml), dried over MgSO4. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with EtOAc/Hexanes (0-60%) gave desired tert- butyl (7-(bis((2-(trimethylsilyl)ethoxy) methyl)amino)-6-bromo-3-(6-fluoroquinolin-3- yl)pyrazolo[1,5-a]pyrimidin-5-yl)metliyl(l-(hydroxymethyI)cyclopropyl)carbamate (O.l9g, 95%).
Figure imgf000604_0001
To tert-butyl (7-(bis((2-(trimethylsiIyl)ethoxy)methyl)amino)-6-bromo-3-(6-fluoroquinolin-3- y i )py razolo [ 1 ,5-a] py rimidin-5-y I )methy 1 ( 1 -(hy droxy methy I ) cyclopropy l)carbamate (0.19g, 0.23mmol) in DCM (2.5ml) was added protonsponge (70mg, 0.33mmol) followed by trimethyloxoniumtetrafluoroborate (48.2mg, 0,33mmol) at room temperature and the resulting mixture was stirred at room temperature for 16 h, at which time LC/MS confirmed full conversion of starting material to product. Reaction mixture was quenched with sat. NH4Cl and diluted with DCM (25ml). Organics were separated and washed with brine (5ml) and dried over MgSO4. Solvent was removed in vacuo and the residue was was purified by column chromatography on silica gel. Elution with DCM/MeOH (0-50%) gave slightly impure desired tert-butyl (7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-6-bromo-3-(6-fluoroquinolin-3- yOpyrazoloU^-alpyrimidin-5-yl)methyKHmethoxymethyOcyclopropyOcarbamate (97mg, 50%).
Figure imgf000604_0002
By essentially following the the same procedure above compounds prepared. LCMS: 1.87 mins, m/z = 473.0 (MH+).
Synthesis of 4-(methoxymethy0tetrahydro-2H-pyran-4-amine:
Synthesis of benzyl 4-(methoxymethyl)tetrahydro-2H-pyran-4-ylcarbamate:
Figure imgf000605_0001
To benzyl 4-(hydroxymethyl)tetrahydro-2H-pyran-4-ylcarbamate (0.59g, 2.23mraol) in DCM (24ml) was added protonsponge (0.53g, 2.5mmol) followed by trimethyloxoniumtetrafluoroborate (0.36g, 2.5mmol) at 0°C. After 20min at 0°C, reaction mixture was warmed up to room temperature and stirred further for 16 h, at which time LC/MS confirmed full conversion of starting material to product. Reaction mixture was quenched with sat. NH4Cl and with DCM (75ml). Organics were separated and washed with brine (20ml) and dried over MgSO4. Solvent was removed in vacuo and the residue was was purified by column chromatography on silica gel. Eiution with EtOAc/Hexanes (66%-i00%) gave slightly impure desired benzyl 4-(methoxymethyl)tetrahydro-2H-pyran-4-ylcarbamate (0.3 Ig, 50%).
Synthesis of 4-(methoxymethyDtetrahydro-2H-pyran-4-amine:
Figure imgf000605_0002
A mixture of benzyl 4-(methoxymethyl)tetrahydro-2H-pyran-4-ylcarbamate (0.86g, 3.1mmol), 10% Pd/C (0.2I g) in EtOAc (25ml) was stirred at room temperature under hydrogen (balloon pressure) for 16 hours. It was then filtered and concentrated to get the desired 4- (m ethoxy methy l)tetrahy dro-2H-py ran-4-am ine.
Figure imgf000606_0001
By essentially the same procedure as to compounds above is prepared. LCMS: 2.173 mins, m/z = 515.2 (MH+).
Synthesis of 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazolof 1 ,5-a]pyrimidine-5- carbonitrile:
Figure imgf000606_0002
5-chloro-N,N-bis((2-(trimethylsiIyl)ethoxy)methyl)pyrazolo[i,5-a]pyrimidin-7-arnine (1.5 g, 3.5 mmol), tributyltin cyanide (1.7 g, 5.3 mmol), tetrakis (triphenylphosphine) palladium (0.8 g, 0.7 mmol), bis(tri-^-butylphosphine) palladium (0.4 g, 0.7 mmol) were charged in a pressure tube. The tube was evacuated and charged with argon for three cycles. Dioxane (20 ml) was added, and the tube was capped and heated at 150°C with stirring for one hour. After cooling, the mixture was diluted with EtOAc (100 ml) and washed with brine (1 x 20 ml), dried over MgSO4. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with EtOAc/Hexanes (0-35%) gave desired product, 7- (bis((2-(trimethylsilyl) ethoxy)methyl)amino) pyrazolo[ l,5-a]pyrimidine-5-carbonitrile (1.3 g, 90%).
Synthesis of 5-π-aminoethyl)-N,N-bis((2~(trimethyisilyl)ethoxy)methyl) pyrazoloM ,5- a]pyrimidin-7-amine:
Figure imgf000607_0001
To a solution 7-(bis((2-(tiimethylsilyl)ethoxy)methyi)amino)pyrazolo[l ,5-a]pyrimidine-5- carbonitrile (0.6 g, i .4 mmol), copper bromide (42 mg, 0.3 mmol), and TBSCl (0.2 g, 1.4 mmol) in THF (14 m!) at O°C was added methyl magnesium bromide (3M in ether, 0.47 ml, 1.4 mmol) dropwise. Reaction mixture was warmed up to room temperature and stirred further for 6 h. LC/MS showed no starting material (M.W = 419) remaining. A borane.THF solution (IM in THF, 2.8 ml, 2.8 mmol) was then added to the reaction mixture at -78°C, and the resulting solution was stirred for 4 h and allowed to warm at room temperature overnight. Distilled water (0.5 ml) was cautiously added to the solution cooled at -20°C, followed by sat. NH4Cl (0.5 ml). After the THF was rotoevaporated, the aqueous phase was extracted with DCM (60 ml), washed with water (1 x 5 ml), brine (1 x 5 ml), dried over MgSO4. Solvent was removed in vacuo and the crude was used for the next step without any further purification.
Alternatively, the 5-(l-aminoethyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl) pyrazolo[l,5- a]pyrimidin-7-amine can also be synthesized by the following procedure.
Synthesis of 5-(1 -aminoethyl)-N,N-bis(f2-(trimethylsilyl)ethoxy)methyl) pyrazolof 1 ,5- alpyrimidin-7-amine can also be synthesized by the following procedure:
MeMgBr
Figure imgf000607_0002
Part A Synthesis of 1 -(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazoloM .5-aipyrimidin- 5-vOethanone:
An oven-dried 250 rriL 3-neck round-bottom flask fitted with a pressure-equalizing addition funnel, temperature probe and septum was charged with 7-(bis{[2- (trimemyisilyI)ethoxy]methyl}amino)pyrazolo[l,5-a]pyrimidine-5-carbonitrile (7.10 g, 16.9 mmol) in dry THF (59 niL), copper© bromide (120 mg, 0.84 mmol) and tert- butyldimethylsilyl chloride (2.80 g, 18.6 mmoi), and the stirred mixture was cooled in an ice- brine bath. 3M MeMgBr in ether (9.59 mL, 28.8 mmol) was added dropwise over 10 min via the addition funnel, and the resulting heterogeneous mixture was stirred at -5-0 °C for 1.5 h and quenched carefully by the dropwise addition of water until gas evolution ceased. The mixture was diluted with additional water (100 mL) and extracted with diethyl ether (2 x 150 mL), and the combined organic phase was washed with brine (100 mL), dried over Na2SO4 and concentrated under reduced pressure. Purification by chromatography on silica gel (120 g), eluting with 0-20% EtOAc/hexanes, afforded the title compound, l-(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)pyrazoIo[1,5-a]pyrimidin-5-yl)ethanone (4.34 g, 59% yield) as a light yellow oil: MS (ESI+) for C20H36N4O3Si2 m/z 437 (M+H)+; R/= 0.70 (25% EtOAc/hexanes).
Part B Synthesis of 1 -(7-(bis((2-(trimethylsilyl)ethoxy)methy0amino)pyrazolof 1 ,5-aipyrimidin- 5-yl)ethanone oxime:
A solution of l-[7-(bis{[2-(trimethylsilyl)ethoxy]methyl }amino)pyrazolo[ 1 ,5-a]pyrimidin-5- yi]ethanone (4.34 g, 9.94 mmol) in 1:1 EtOH/pyridine (50 mL) was treated with hydroxyiamine hydrochloride (3.45 g, 49.7 mmol). The resulting mixture was stirred at room temperature for 1.5 h and then concentrated under reduced pressure to remove most solvent. The residue was partitioned between EtOAc (100 mL) and water (60 mL), the layers were separated, and the organic phase was washed with water (60 mL) and brine (20 mL), dried over Na2SO4 and concentrated under reduced pressure to give the title compound, l-(7-(bis((2- (trimethylsilyl)ethoxy)methyI)amino)pyrazolo[l,5-alpyrimidin-5-yl)ethanone oxime (4.90 g, 85% purity, 93% yield) as an off-white semisolid: MS (ESI+) for C20H37N5O3Si2 m/z 452 (M+H)+.
Part C Synthesis of 5-(1-aminoethyi)-N.N-bisf(2-(trimethylsilyS)ethoxy)methyl)pyrazolori ,5- aipyrimidin-7-amine:
: A solution of l-[7-(bis{[2-(trimethylsily0ethoxy]methyl}amino)pyrazolo[l,5-a]pyrimidin-5- yl]ethanone oxime (4,50 g, 8.77 mmol) in 4: 1 EtOH/ AcOH (90 mL) under nitrogen in a Parr bottle was treated with 10% Pd on carbon (933 mg, 0.877 mmol), and the mixture was shaken on the Parr apparatus under 45 psi hydrogen for 23 h. The catalyst was removed by filtration through CeI ite, and the filtrate was concentrated under reduced pressure, azeotroping with toluene at 40-45 °C to remove most AcOH. The oily residue was taken up in EtOAc (-100 mL), washed with saturated aqueous NaHCO3 (2 x 50 mL) and brine (20 mL), dried over Na2SO4, and concentrated under reduced pressure to give the title compound, 5-(l- aminoethyl)-N,N-bis((2-(tximethylsilyl)ethoxy)methyl)pyrazolo[l ,5-a]pyrimidin-7-amine (4.02 g, 91% purity, 95% yield) as an oil: MS (ESI+) for C20H39N5O2Si2 m/z 438 (M+H)+.
Synthesis of 5-(1 -aminoethyl)-3-(6-fiuoroquinolin-3-yl)-N,N-bis((2- (trimethylsiM)ethoxy)methyθpyrazolo[1 ,5~a1pyrimidin-7-amine:
Figure imgf000609_0001
Part A
Synthesis of 1 -(7-(bis(f2-(trimethylsiiyl)ethoxy)methyl)annino)-3-(6-fluoroauinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)ethanol:
To a solution of 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3- yl)pyrazolo[1,5-a]pyrimidine-5-carbaidehyde (11.0 g, 19.4 mmol) in anhydrous THF (200 mL, 2 mol) under nitrogen at 0 °C was added 3M methylmagnesium bromide in ether (9.69 mL, 29.0 mmol) dropwise. After 1 hour the reaction was complete by HPLC. Water (10 mL) was added over a 15 minute period to quench the reaction. Removed the flask from the ice bath and concentrated in vacuo. Dissolved the remaining residue in ethyl acetate, washed three times with water and then twice with brine. The organics were dried over sodium sulfate, filtered and concentrated in vacuo. Purified the remaining crude material by flash chromatography (5-40% EtOAc/hex) to afford l-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3- yl)pyrazolo[l ,5-a]pyrimidin-5-yl)ethanol (6.8 g; 60% yield) as a yellow solid. HPLC: 7.54 min (Method C). LCMS: ESI+ m/z of 584 (M+H)
Part B
Synthesis of 1 -(7-( bis((2-(trimethylsilyl)ethoxy)methyi)amino)-3-(6-f luoroquinoiin-3- yl)pyrazolofi ,5-a1pyrimidin-5-yl)ethyl methanesulfonate:
To a solution of l-(7-(bis((2-(trimethylsilyl)ethoxy)methy!)amino)-3-(6-fIuoroquinolin-3- yl)pyrazolo[1,5-a]pyrimidin-5-yl)ethanol (6.1 g, 10.4 mmol) in methylene chloride (100 mL) and triethytamine (7.1 mL, 50.9 mmol) under nitrogen was added methanesulfonyl chloride (2.4 mL, 31.0 mmol) dropwise. After 15 minutes the reaction was complete by HPLC. The contents of the flask were transferred to a separatory funnel, washed twice with water and twice with brine. The organics were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(7-(bis((2-(trimethyIsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3- yl)pyrazoIo[1,5-a]pyrimidin-5-yl)ethyl methanesulfonate (6.8 g; 98% yield) as a dark yellow oil that was used as is for the following step. HPLC: 6.18 min (Method D). LCMS: ESI+ m/z of 662 (M+H)
Part C
Synthesis of 5-( 1 -azidoethyl)-3-(6-fluoroquinolin-3-yl)-N,N-bis((2- (trimethylsiiyl)ethoxy)methy0pyrazolof1 ,5-a1pyrimidin-7-amine:
To a solution of l-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinoiin-3- y!)pyrazolo[1,5-a]pyrimidin-5-yl)ethyl methanesulfonate (6.8 g, 10.3 mmol) in DMSO (100 mL) and N,N-diisopropyl-ethylamine (8.95 mL, 51.4 mmol) was added sodium azide (2.0 g, 30.8 mmol). After 4 hours the reaction was complete by HPLC. The contents of the flask were transferred to a separatory funnel, washed twice with water and twice with brine. The organics were dried over sodium sulfate, filtered and concentrated in vacuo to afford 5-(l -azidoethyl)-3- (6-fluoroquinolin-3-yl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7- amine (6.2 g; 99% yield) as a dark yellow oil. HPLC: 6.67 min (Method D). LCMS: No ion detected.
Part D
Synthesis of 5-(1 -aminoethyπ-3-(6-fluoroαuinolin-3-yl)-N,N-bisf (2- ftrimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-alpyrimidin-7-amine:
To a solution of 5-(l-azidoethyl)-3-(6-fluoroquinolin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (6.2 g, 10.3 mmol) and THF (90 mL) was added IM trimethylphosphine in THF (30.8 mL, 30.8 mmol) dropwise. After 30 minutes the reaction was complete by HPLC. Water (30 mL) was therefore added and the reaction stirred an additional hour. The contents of the flask were transferred to a separatory funnel, diluted with 300 mL of EtOAc and washed three times with water and three times with brine. The organics were dried over sodium sulfate, filtered and concentrated in vacuo. Purified the remaining crude material by flash chromatography (0-2% 0.7N NH3 in MeOH/CHC13) to afford 5-(l -aminoethyl)-3-(6-fluoroquinolin-3-yl)-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (4.9 g, 82% yield) as a yellow solid. HPLC: 6.62 min (Method C). LCMS:ESI+ m/z of 583 (M+H)
Synthesis of 5-( 1 -(tetrahydro-2H-Dyran-4-ylarπino)ethyπ-N.N-bis(f2-(trimethy!silyl) ethoxy)methyl)pyrazoloπ .5-aipyrimidin-7-amine:
Figure imgf000611_0001
To crude 5-(l-aminoethyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5- a]pyrimidin-7-amine (0.6 g) in DCE (20 ml) and Ethanol (20 ml) was added tetrahydro-4H- pyran-4-one (0.6 ml, 7 mmol) followed by AcOH (0.08 ml, 1.4 mmol). After stirring for 30 minutes at room temperature, NaBH3CN (0.18 g, 2.8mmoi) was added and stirring was continued further for 30 minutes more. LC/MS showed no starting material (M.W = 437) remaining. After the solvent was rotoevaporated, the crude was dissolved with DCM (60ml), washed with sat. NaHCO3 (1 x 5ml), water (1 x 5ml), brine (1 x 5ml), dried over MgSO4. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Eiution with MeOH/EtOAC (0-20%) gave desired product, 5-(l -(tetrahydro-2H- pyran-4-ylamino)ethyl)-N,N-bis((2-(trimethylsilyl) ethoxy)methyl) pyrazolo[l ,5-a]pyrimidin- 7-amine (0.2 g, 27%).
Synthesis of 3-iodo-5-(1-(tetrahydro-2H-pyran-4-ylamino)ethyl)-N,N-bis((2- (trimethylsiivhethoxy)methyl)pyrazolof1 ,5-a]pyrimidin-7-amine:
Figure imgf000611_0002
To a solution of 5-(l-(tetrahydro-2H-pyran-4-ylamino)ethyl)-N,N-bis((2-(trimethylsiIy[) ethoxy) methyl) pyrazolo[1,5-a]pyrimidin-7-amine (0.17g, 0.32mmol) in acetonitrile (15 ml) at 0°C was added NIS (78mg, 0.35 mmol) and stirring continued for 10 minutes. LC/MS showed no starting material (M.W = 521) remaining. Saturated sodium thiosulfate solution (~0.5ml) was added and stirring continued for 5 minutes. After the acetonitrile was rotoevaporated, the aqueous phase was extracted with DCM (25ml), washed with water (I x 5ml), brine ( I x 5ml), dried over MgSO4. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with MeOH/EtOAC (0-20%) gave desired product, 3- iodo-5-(l-(tetrahydro-2H-pyran-4-ylamino)ethyl)-N,N-bis((2- (trimethylsilyl)ethoxy)methy[)pyrazolo[ 1 ,5-a]pyrimidin-7-amine (0.14g, 69%).
Synthesis of 3-(6-fluoroQuinolin-3-yl)-5-(1-(tetrahydro-2H-pyran-4-ylamino)ethyh-N .N- bis(f2-(trimethylsilyl)ethoxy)methyl)pyrazolori ,5-aipyrimidin-7-amine:
Figure imgf000612_0001
To 3-iodo-5-( 1 -(tetrahydro-2H-pyran-4-ylamino)ethyl)-N,N-bis((2-(trimethylsilyl) ethoxy)methyl)pyrazolo[1,5-a]pyrimidin-7-amine (0.14g, 0.21mmol) was added 6- fluoroquinoline boronate (0.1 Ig, 0.42mmol), K2CO3 (87mg, 0.63mmol), PdC!2(dppf).CH2Cl2 (18mg, 0.02mmoi), dioxane (4ml) and H2O (ImI). The resulting mixture was heated at 100 °C for 2 hours, at which time LC/MS analysis confirmed full consumption of starting material. On cooling, the solvent was rotoevaporated, and the crude was redissoived in DCM (40ml), washed with water (1 x 5ml), brine (i x 5ml), dried over MgSO4. Solvent was removed in vacuo and the residue was purified by column chromatography on silica gel. Elution with MeOH/EtOAC (0-20%) gave desired product, 3-(6-fluoroquinolin-3-yl)-5-( 1 -(tetrahydro-2H- pyran-4-ylamino)ethyl)-N,N-bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[l,5-a]pyrimidin-7- amine (84mg, 60%).
Synthesis of 3-(6-f luoroquinolin-3-vh-5-(1 -(tetrahydro-2H-pyran-4- ylamino)ethyl)pyrazolof1 ,5-alpyrimidin-7-amine:
Figure imgf000613_0001
To a solution of 3-(6-fiuoroquinolin-3-yl)-5-(l-(tetrahydro-2H-pyran-4-ylamino)ethyl)-N,N- bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[ l,5-a]pyrimidin-7-amine (84 mg, 0.13mmol) in TFA (2 ml) was added few drops of water and stirring continued for 2h at room temperature. LC/MS showed no starting material (M. W = 666) remaining. TFA along with water was rotoevaporated, and the crude was dried under the high vacuum for 4 h, which was used without further purification for the next step.
Synthesis of 6-bromo-3-(6-fluoroQuinolin-3-yl)-5-(1 -(tetrahydro-2H-pyran-4- ylamino)ethyl)pyrazolo[1 ,5-aipyrimidin-7-amine:
Figure imgf000613_0002
To a solution of 3-(6-fluoroquinolin-3-y!)-5-(l-(tetrahydro-2H-pyran-4- ylamino)ethyl)pyrazolo[1,5-a]pyrimidin-7-amine (Crude, ~0.13mmol) in DMF (2ml) at room temperature was added NBS (25mg, 0.14mmol) and stirring continued for 10 minutes. LC/MS showed no starting material (M.W = 406) remaining. This crude compound was submitted to the analytical group for purification to afford the desired 6-bromo-3-(6-fluoroquinolin-3-yl)-5- (l-(tetrahydro-2H-pyran-4-yIamino)ethyi)pyrazolo[1,5-a]pyrimidin-7-amine.
Synthesis of H4-aminopiperidin-1-yl)-2-methoxyethanone:
Figure imgf000614_0001
Methyl chloroformate (289 μl_, 3.74 mmol) was added to a solution of 4-(N-Boc- amino)-piperidine (500 mg, 2.50 mmol) in methylene chloride (25 mL) at room temperature. N,N-diisopropylethy!amine (1.30 mL, 7.49 mmol) was added and the solution was stirred overnight. The reaction mixture was diluted with sat. ammonium chloride (30 mL) and extracted with methylene chloride (3 x 30 mL). The combined extracts were dried over sodium sulfate and concentrated. The crude material was purified by column chromatography on silica gel (0% to 100% ethyl acetate in methylene chloride) to give 566 mg of a white soiid. This solid was taken up in methylene chloride (2 mL) and trifluoroacetic acid (2 mL) was added at room temperature. After 20 min, the solution was concentrated, and then diluted with methylene chloride (2 mL). The solution was basified with sat. sodium carbonate (2 mL) and the mixture was extrated with methylene chloride (5 x 5 mL) and ethyl acetate (5 x 5 mL). The combined extracts were concentrated and the resulting soiid was freeze dried from acetonitrile and water to give 290 mg of the title compound as a white solid, 1 -{4-aminopiperidin-1-yl)-2-methoxyethanone, (73% over two steps). NMR (300 MHz, D2O) δ 4.79 (s, 2H), 4.12 (d, J= 13.5 Hz, 2H), 3.72 (s, 3H), 3.28-3.14 (m, 1 H), 2.95 (t, J = 12.6 Hz1 2H), 1.98 (d, J = 12.0 Hz1 2H)1 1.54-1.36 (m, 2H).
Synthesis of 1-(4-aminopiperidin-1-yl)-3-methoxypropan-1-one:
Figure imgf000614_0002
3-Methoxypropanoic acid (225 μL, 2.40 mmol) was added to a solution of 4-(N-Boc- amino)-piperidine (400 mg, 2.00 mmol) in methylene chloride (25 mL) at room temperature. N,N-dsisopropylethylamine (1.30 mL, 7.49 mmol) and propylphosphonic anhydride (1.78 mL, 50 wt. % in ethyl acetate, 3.00 mmol) was added, and the solution was stirred for 4.5 h. The reaction mixture was concentrated and partitioned between ethyl acetate (5 mL) and water (5 mL). The organic portion was washed with 0.2M HCl (2 mL), sat. Sodium bicarbonate (5 mL) and brine (5 mL). The solution was then dried over sodium sulfate and concentrated to a white solid. This solid was taken up in methylene chloride (6 mL), and trifluoroacetic acid (3 mL) was added at room temperature. After 1 h the solution was concentrated. The resulting solid was dissolved in methylene chloride (2 mL) and 1 M sodium hydroxide (1 mL). This mixture was extracted with methylene chloride (4 x 2 mL) and ethyl acetate (4 x 2 mL). The combined organic extracts were dried over sodium sulfate and concentrated to give 135 mg of the title compound, 1 -(4-aminopiperidin-1-yl)-3-methoxypropan-1-one (36% over two steps). 1H NMR (300 MHz, CD3OD) δ 4.51-4.41 (m, 1H), 4.03-3.93 (m, 1 H), 3.64 (d, J = 6.3 Hz, 2H), 3.32 (s, 3H), 3.17-3.05 (m, 1 H), 2.96-2.83 (m, 1 H), 2.75-2.55 (m, 3H), 1.99-1.80 (m, 2H), 1.40-1.14 (m, 2H).
Synthesis of (R)-1-(4-aminopiperidin-1-yl)-2,3-dihydroxy-3-methyibutan-1 -one:
Figure imgf000615_0001
3,3-Dimethylacryioyl chloride (670 μl_, 5.99 mmol) was added to a solution of 4-(N- Boc-amino)-piperidine (1.00 g, 4.99 mmol) in methylene chloride (50 mi_) at room temperature. N,N-diisopropylethylamine (1.74 ml_, 9.99 mmol) was added and the solution was stirred overnight. The reaction mixture was diluted with sat. ammonium chloride (50 rriL) and extracted with methylene chloride (3 x 50 rriL). The combined extracts were dried over sodium sulfate and concentrated. The crude material was purified by column chromatography on silica gel (0% to 100% ethyl acetate in methylene chloride) to give 1.39 g of an off white solid. This solid was dissolved in t- butanol (18 mL) and water (18 ml_), and the solution was cooled to 0 °C. To this mixture were added AD-mix-α (7.50 g), methanesulfonamide (449 mg, 4.72 mmol), sodium bicarbonate (1.13 g, 13.5 mmol}, and potassium osmate dihydrate (15 mg? 0.0450 mmol). The solution was allowed to warm to room temperature overnight. A 1.9M solution of sodium sulfite (40 mL) was added and the mixture was stirred for 1 h. The resulting aqueous solution was extracted with ethyl acetate (4 x 10OmL) and dried over sodium sulfate. The solution was concentrated and the crude material was purified by column chromatography on silica gel (0% to 20% methanol in methylene chloride, and 0% to 10% methanol in methylene chloride) to give 670 mg of the diol. A portion of this diol (650 mg, 2.05 mmol) was dissolved in dioxane (10 mL) and HCl in dioxane (5.1 mL, 4M, 20.5 mmol) was added at room temperature. After stirring overnight, the reaction mixture was diluted with ether (20 mL) and filtered to give a white solid. This solid was taken up in methylene chloride (50 mL) and MP-carbonate resin (830 mg, 2.11 mmol) was added. After 1.5 h the solvent was decanted and the resin was washed with methylene chloride (20 mL). Methylene chloride (50 mL) was added to the resin along with additional fresh resin (1.0 g, 2.54 mmol). After stirring for 30 min, the solvent was decanted andthe remaining resin was washed with methylene chloride (20 mL). The combined organics were concentrated to 121 mg of the free amine, (R)-1 -(4-aminopiperidin-1-yl)-2,3-dihydroxy-3-methylbutan-1 -one (13% over three steps). 1H NMR (300 MHz, CD3OD) δ 4.54-4.40 (m, 1 H), 4.32-4.26 (m, 1 H)1 4.26-4.13 (m, 1 H), 3.16-3.03 (m, 1 H), 2.95-2.81 (m, 1 H), 2.80-2.69 (m, 1 H), 1.94-1.79 (m, 2H), 1.47-1.28 (m, 2H), 1.25 (d, J = 5.4 Hz, 3H), 1.17 (d, J = 5.1 Hz, 3H). Synthesis of (R)-1 -(4-aminopiperidin-1-yl)-2,3-dihydroxypropan-1 -one:
Figure imgf000616_0001
N.N-diisopropylethylamine (2.54 mL, 14.6 mmol) was added to a solution of 4-(N-Boc- amino)-piperidine (1.75 g, 8.75 mmol) in tetrahydrofuran {36 mL) at O °C. Propylphosphonic anhydride (5.92 mL, 50 wt. % in ethyl acetate, 9.48 mmol) and acrylic acid (500 μL, 7.29 mmol) were added, and the solution was allowed to warm to room temperature overnight. The reaction mixture was partitioned between ethyl acetate (20 mL) and water (20 mL). The organic portion was washed with 0.2M HCI (5 mL), sat. sodium bicarbonate (20 mL), and brine (20 mL). The solution was then dried over sodium sulfate and concentrated to a white solid. A portion of this solid (800 mg, 3.15 mmol) was dissolved in f-butanol (13 mL) and water (13 mL) and cooled to 0 °C. To this mixture were added AD-mix-α (5.24 g), methanesulfonamide (314 mg, 3.30 mmol), sodium bicarbonate (793 mg, 9.44 mmol), and potassium osmate dihydrate (52 mg, 0.157 mmol). After 4.5 h, a 0.8M solution of sodium sulfite (100 mL) was added and the mixture was stirred for 1 h at room temperature. The resulting aqueous solution was extracted with ethyl acetate (4 x 10OmL) and dried over sodium sulfate. The solution was concentrated and the crude material was purified by column chromatography on silica gel (0% to 20% methanol in methylene chloride) to give a clear syrup. A portion of this syrup (790 mg, 2.74 mmol) was dissolved in dioxane (14 mL) and HCI in dioxane (6.8 mL, 4M, 27.4 mmol) was added at room temperature. After 18 h, the solution was concentrated. The resulting white solid was taken up in methylene chloride (50 mL) and MP-carbonate resin (3.2 g, 8.1 mmoi) was added. After stirring for 30 min, the solvent was decanted. The resin was washed with ethyl acetate (10 mL) and methylene chloride (10 mL), and the combined organics were discarded. The remaining resin was diluted with methanol (50 mL) and stirred for 30 min. The methanol was decanted and the resin was washed with methanol (10 mL). The combined methanol portions were concentrated to give clear oil, (R)-1 -(4- aminopiperidin-1-yl)-2,3-dihydroxypropan-1-one, 382 mg (60% over three steps). 1H NMR (300 MHz, CD3OD) δ 4.53-4.40 (m, 2H), 4.14-4.04 (m, 1 H), 3.77-3.56 (m, 2H), 3.20-3.05 (m, 1 H), 3.04-3.92 (m, 1 H), 2.84-2.69 (m, 1 H), 2.00-1.82 (m, 2H), 1.48-1.20 (m, 2H).
Synthesis of (4-aminopiperidin-1 -yl)(3-methyloxetan-3-yl)methanone:
Figure imgf000617_0001
A solution of 3-hydroxymethyI-3-methyloxetane (1.90 mL, 19.1 mmol) in methylene chloride (6 mL) was added to a solution of nickel (II) chloride hexahydrate (113 mg, 0.476 mmol) in water (2 mL). This mixture was cooled to 0 °C and bleach (127 mL) was added over a 30 min period via addition funnel. After 2 h the ice bath was removed and the reaction mixture was stirred at room temperature for an additional 2 h. 2M HCI (120 mL) was added and the solution was extracted with ether (4 x 200 mL). The combined extracts were dried over sodium sulfate and concentrated to a clear oil. A portion of this oil (1.00 g, 8.61 mmol) was dissolved in tetrahydrofuran (43 mL) and 4-(N-Boc-amino)-piperidine (2.07 g, 10.3 mmol) was added at room temperature. Propylphosphonic anhydride (6.99 mL, 50 wt. % in ethyl acetate, 11.2 mmol) and N,N-diisopropylethylamine (3.00 mL, 17.2 mmol) were added, and the reaction mixture was stirred for 24 h. The solution was partitioned between ethyl acetate (35 mL) and water (35 mL). The organic portion was washed with 0.2M HCI (10 mL), sat. sodium bicarbonate (35 mL), and brine (35 mL). The solution was then dried over sodium sulfate and concentrated. The crude material was purified by column chromatography on silica gel (0% to 100% methylene chloride in ethyl acetate). The resulting amide (213 mg, 0.714 mg) was diluted with methylene chloride (7 mL) and trifluoroacetic acid (1.8 mL) was added at room temperature. After 2 h the solution was concentrated and a 2M sodium hydroxide solution was added. The aqueous solution was extracted with ethyl acetate (3 x 10 mL). The combined extracts were dried over sodium sulfate and concetrated to give the title compound as a clear oil, (4-aminopiperidin-1-yl)(3-methyloxetan-3-yl)methanone, 68 mg (4% over three- steps). 1H NMR (300 MHz, CD3OD) δ 4.95-4.88 (m, 2H), 4.48-4.38 (m, 1 H), 4.38-4.31 (m, 2H), 3.19-2.96 (m, 2H), 2.94-2.81 (m, 1 H), 2.81-2.66 (m, 1 H), 1.93-1.81 (m, 2H), 1.67-1.60 Jm1 3H)1 1.36-1.15 (m, 2H).
Synthesis of (2R,3S)-1 -(4-aminopiperidin-1-yl)-2,3-dihydroxybutan-1~one: 1. crotyl chloride
Figure imgf000618_0001
3. HCI1 dioxane, rt
Crotyl chloride (674 μl_, 5.99 mmol) was added to a solution of 4-(N-Boc-amino)- piperidine (1.00 g, 4.99 mmol) in methylene chloride (50 ml_) at 0 °C. N, N- diisopropylethylamine (1.74 mL, 9.99 mmol) was added and the solution was stirred for 21 h. The reaction mixture was diluted with sat. ammonium chloride (50 mL) and extracted with methylene chloride (3 x 50 mL). The combined extracts were dried over sodium sulfate and concentrated. The residue was dissolved in methylene chloride (10 mL) and was triturated with hexanes (100 mL). The solvent was filtered off to give an off white solid. This solid was dissolved in f-butanoi (16 mL) and water (16 mL) and cooled to 0 °C. To this mixture were added AD-mix-α (6.83 g), methanesuifonamide (409 mg, 4.30 mmol), sodium bicarbonate (1.03 g, 12.3 mmol), and potassium osmate dihydrate (68 mg, 0.205 mmol). The solution was allowed to warm to room temperature overnight. A 0.5M solution of sodium sulfite (100 mL) was added and the mixture was stirred for 1 h. The resulting aqueous solution was extracted with ethyl acetate (4 x 10OmL) and dried over sodium sulfate. The solution was concentrated and the crude material was purified by column chromatography on silica gel (0% to 20% methanol in methylene chloride, and 0% to 20% methanol in methylene chloride) to give the diol as a white solid. A portion of the diol (450 mg, 1.49 mmol) was dissolved in dioxane (7 mL) and HCI in dioxane (3.7 mL, 4M, 27.4 mmol) was added at room temperature. After 1.5 h, the solution was concentrated. The resulting white solid was taken up in methylene chloride (30 mL) and MP-carbonate resin (2.0 g, 5.1 mmol) was added. After stirring for 45 min, the solvent was decanted, and the resin was washed with methylene chloride. The remaining resin was washed with methanol (2 x 30 mL) and the combined methanol portions were concentrated to give a clear oil, (2R,3S)-1 -(4-aminopiperidin-1-yl)-2,3-dihydroxybutan-1 -one, 160 mg (33% over three steps). 1H NMR (300 MHz, CD3OD) 54.51-4.35 (m, 1 H), 4.32-4.25 (m, 1H), 4.14-4.01 (m, 1H), 3.96-3.84 (m, 1 H), 3.19-3.02 (m, 1 H), 2.98-2.84 (m, 1 H), 2.84-2.67 (m, 1 H), 1.97-1.80 (m, 2H), 1.44-1.20 (m, 2H), 1.18 (s, 3H).
Synthesis of 1-(4-amino-4-methylpiperidin-1 -yl)ethanone:
Figure imgf000618_0002
Methylene chloride (53 ml_) was added to a mixture of 4-amino-4- methylpiperidine»2HCI (500 mg, 2.67 mmol) and 4-(dimethy!amino)pyridine (33 mg, 0.267 mmol). N,N-diisopropylethylamine (4.65 ml_, 26.7 mmol) was added and the mixture was cooled to -78 °C. Acetic anhydride (126 μL, 1.34 mmol) was added and the solution was allowed to warm to room temperature overnight. 2M sodium hydroxide (10 rnL) was added and the mixture was extracted with ethyl acetate (5 x 20 ml_) and methylene chloride (5 x 20 mL). The combined organic extracts were dried over sodium sulfate and concentrated. The crude material was purified by column chromatography on silica gei (0% to 20% methanol in methylene chloride w/ 0.1% ammonium hydroxide) to give 181 mg of the title compound, 1-(4-amino-4- methylpiperidin-1 -yl)ethanone (87%). 1H NMR (300 MHz, CD3OD) δ 3.67-3.40 (m, 4H), 2.08 (s, 3H), 1.64-1.38 (m, 4H), 1.18 (s, 3H).
Synthesis of 1 -(4-aminopiperidin-1 -yl)-3-hydroxy-3-methylbutan-1 -one:
Figure imgf000619_0001
3-hydroxy-3-methyl butyric acid (1.20 equiv), DIPEA (2.00 equiv), and T3P (1.50 equiv) were added to a stirring solution of compound 1 (1.00 equiv) in THF (0.2M) at room temperature under nitrogen. TLC analysis after 2 hours showed almost complete conversion, so reaction was concentrated. Residue was dissolved in ethyl acetate then washed with water, dilute HCI, saturated NaHCO3, and brine. The phases were separated and the organics dried over sodium sulfate, filtered, and concentrated. The resulting residue was purified by flash column chromatography (SiO2, 12 g; 0% to 10% methanol in methylene chloride). The resulting material was dissolved in methylene chloride (0.97M) and TFA (0.97M) was added under nitrogen and the mixture was stirred at room temperature for 1 hour at which time HPLC analysis indicated the reaction was complete. Reaction was concentrated and the resulting residue was dissolved in 1 N NaOH and stirred at room temperature for 30 minutes. The aqueous mixture was extracted with methylene chloride. The phases were separated and the organics dried over sodium sulfate, filtered, and concentrated affording the title compound, 1 -(4-aminopiperidin-1 -yl)-3-hydroxy-3-methylbutan-1 - one. 1H NMR (300 MHz1 CDCI3) δ 4.40 (d, 1H, JHz=13.9), 3.73 (d, 1 H, JHz=12.1 ), 2.99-2.95 (m, 1H), 2.90 (m, 1 H), 2.70-2.63 (m, 1 H), 2.33 (s, 2H), 1.76 (m, 2H), 1.12 (s, 8H). General synthesis of 1-(4-aminopiperidin~1-yl)-acyl derivatives:
Figure imgf000620_0001
DlPEA (3.00 equiv) and propionyl chloride {compound A), isopropyl sulfonyl chloride (compound B), dimethyl carbamyl chloride (compound C), or isobutyryl chloride (compound D) (1.50 equiv) were added to a stirring solution of tert-butyl piperidin-4- ylcarbamate (1.00 equiv) in methylene chloride (0.1 M) at 0 °C. Reaction warmed to room temperature. TLC analysis after 16 hours showed no remaining starting materia! and reaction was diluted in saturated NH4CI and extracted with methylene chloride. The phases were separated and the organics dried over sodium sulfate, filtered, and concentrated. The resulting residue was purified by flash column chromatography (SiO2, 40 g; 0% to 100% ethyl acetate in hexanes). The resulting material was dissolved in methylene chloride (1.0M) and TFA (1.0M) was added under nitrogen and the mixture was stirred at room temperature for 30 minutes at which time TLC analysis indicated the reaction was complete. Reaction was concentrated and the resulting residue was dissolved in 1 N NaOH and stirred at room temperature for 30 minutes. The aqueous mixture was extracted with methylene chloride. The phases were separated and the organics dried over sodium sulfate, filtered, and concentrated . affording the title compound.
Compound A: 1H NMR (300 MHz, CDCl3) δ 4.53 (d, 1H, JHz=13.2), 3.84 (d, 1H,
JHz=13.2), 3.11-3.02 (m, 1 H), 2.97-2.87 (m, 1H), 2.76-2.67 (m, 1 H), 2.40-2.33 (m, 2H), 1.89-1.83 (m, 2H)5 1.37-1.14 (m, 5H);
Compound B: 1H NMR (300 MHz, CDCI3) δ 3.82-3.77 (m, 2H), 3.26-3.12 (m, 1 H)1 3.02-2.93 (m, 2H), 2.90-2.81 (m, 1 H), 1.91-1.86 (m, 2H)1 1.47-1.33 (m, 8H); Compound C: 1H NMR (300 MHz, CDCi3) δ 3.54 (d, 2H1 JHz=14.1 ), 2.77-2.65 (m, 7H), 1.75-1.70 (m, 2H), 1.30-1.61 (m, 4H);
Compound D: 1H NMR (300 MHz, CDCI3) δ 4.50-4.55 (d, J= 13.5 Hz, 1H), 3.89-3.92 (d, J = 12.3, 1 H), 3.03-3.11 (t, J= 12.2 Hz, 1H), 2.76-2.95 (m, 2H), 2.64-2.73 (t, J = 12.9 Hz, 1 H) 1.82-1.90 (m, 2H), 1 .32 (s, 2H), 1.19-1.32 (m, 2H), 1.11 -1.13 (d, J= 6.3 Hz1 6H). General Synthesis of 3-aryl-6-bromo-5-(1 -(disubstitutedamino)ethyQpyrazolori .5- a]pyrimidin-7-amine:
Scheme-XXXXXX
Figure imgf000621_0001
Synthesis of 3-aryl-5-(1-(disubstitutedamino)ethyπ-N,N-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazolori ,5-aipyrimidin-7-amine:
Sodium cyanoborohydride (3.5 equiv) was added to a stirring mixture of ketone, 1 -(3- argio-7-(bis{(2-(trimethylsiiyl)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)ethanone (1 equiv) and amine (2 equiv} in ethanol and acetic acid at room temperature. When the reaction was deemed complete by HPLC, LC-MS, or TLC analysis, the mixture was quenched with saturated aqueous sodium bicarbonate, then extracted with dichloromethane (2x). The combined organics were dried over sodium sulfate, filtered, concentrated to dryness, and purified on silica gel to afford compound 3-aryl-5-(1-(disubstitutedamino)ethyl)-N,N-bis((2- (trimethylsiIyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine.
Synthesis of 3-Aryl -6-bromo-5-(1 -(disubstitutedamino)ethyl)pyrazolori ,5-a]pyrirnidin- 7-amine: Compound, 3-aryl-5-(1-(disubstitutedamino)ethyl)-N,N-bis((2-
(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidin-7-amine, was stirred in a mixture of TFA, dichloromethane, and water, then concentrated to dryness when the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. This material was then treated with NBS (1.05 equiv) in acetonithle. When the reaction was deemed complete by HPLC, LC-MS, or TLC analysis, the mixture was concentrated to dryness, purified by prep-HPLC, and freeze-dried to afford compound 3-Aryl -6- bromo-5-(1 -(disubstitutedamino)ethyl)pyrazolo[1 ,5-a]pyrimtdin-7-amine.
General synthesis of 3-aryl-6-acetyl-5-π-(disubstitutedamino)ethyθpyrazoiof1.5- aipyrimidin-7-amine:
Figure imgf000622_0001
A mixture of compound 3-ary!-5-(1-(disubstitutedamino)ethyl)-N,N-bis({2- {trimethylsiIyl)ethoxy)methyl)pyrazoio[1,5-a]pyrimidin-7-amine. (1 equiv) and NBS (1.05 equiv) in acetonitrile was stirred at room temperature until the reaction was deemed complete by HPLC1 LC-MS, or TLC analysis. The reaction mixture was concentrated, re-dissolved in dichloromethane, washed with saturated aqueous sodium thiosulfate, dried over sodium sulfate, filtered, concentrated, and purified on silica ge!. The resulting material was stirred with a mixture of vinyl stannane (2 equiv), palladium catalyst (0.1 equiv), and potassium fluoride (3 equiv) in dioxane at 85 °C until the reaction was deemed complete by HPLC1 LC-MS, or TLC analysis. The mixture was allowed to cool to room temperature, diluted with 10% aqueous potassium fluoride, and extracted three times with ethyl acetate. The combined organics were dried over sodium sulfate, filtered, concentrated, and purified on silica gel. The resulting material was stirred with a mixture of HCl in methanol and water at 60 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The mixture was then allowed to cool to room temperature, concentrated, purified by prep-HPLC, and freeze-dried to afford compound 3-aryl-6-acetyl-5-(1 - (disubstitutedamino)ethyl)pyrazolo[1 ,5-a]pyrimidin-7-amine.
4-((7-amino-6-bromo-3-(6-fluoroαuinolin-3-yl)pyrazolo[1 ,5-aipyrimidin-5- ylamino)methyl)-N-alkylpiperidine-1 -carboxamide:
Figure imgf000622_0002
SCHEME-47
Sodium cyanoborohydride (3.5 equiv) was added to a stirring mixture of aldehyde, 7- (bis((2-{trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3-yl)pyrazoio[1 ,5- a]pyrimidine-5-carbaidehyde (1 equiv) and 1-benzylpiperidin-4-yl)methanamine (2 equiv) in ethanol and acetic acid at room temperature. When the reaction was deemed complete by HPLC, LC-MS, or TLC analysis, the mixture was quenched with saturated aqueous sodium bicarbonate, then extracted with dichloromethane (2x). The combined organics were dried over sodium sulfate, filtered, concentrated to dryness, and purified on silica gel to afford the desired product, N5-((1 - benzylpiperidin-4-yl)methyl)-3-(6-fluoroquinolin-3-y!)-N7,N7-bis({2- (trimethylsilyl)ethoxy)methy!)pyrazolo[1 ,5-a]pyrimidine-5,7-diamine.
The N5-{(1-benzylpiperidin-4-yl)methyl)-3-{6-fluoroquinolin-3-yl)-N7,N7-bis((2- (trimethylsilyl)ethoxy)methyl)pyrazoIo[1 ,5-a3pyrimidine-5,7-diamine (1 equiv), BoC2O (2 equiv), DMAP (0.2 equiv), and triethylamine (3 equiv) in dichloromethane was stirred at room temperature until the reaction was deemed complete by TLC, HPLC, or LC- MS analysis. The mixture was diluted with additional dichloromethane, washed with saturated aqueous ammonium chloride, dried over sodium sulfate, filtered, concentrated, and purified on silica gel to give the product, tert-butyl (1- benzylpiperidin-4-yl)methyl(7-(bis({2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- fluoroquinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yi)carbamate.
This material, tert-butyl (1 -benzyipiperidin-4-yl)methyl(7-(bis((2- (trimethylsilyl)ethoxy)methyl)amino)-3-{6-fIuoroquinolin-3-yI)pyrazolo[1 ,5-a]pyrimidin- 5-yl)carbamate, was then stirred with Pd(OH)2/C (0.2 equiv) and ammonium formate (20 equiv) in ethanol at 80 °C until the reaction was deemed complete by HPLC, LC- MS, or TLC analysis. The mixture was allowed to cool to room temperature, filtered through a pad of celite, concentrated and purified on silica gel to afford compound tert- butyl 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl(piperidin-4-ylmethyl)carbamate.
6-bromo-3-(6-fluoroquinolin-3-yl)-N5-(piperidin-4-ylmethyl)pyrazolori,5-aipyrimidine- 5,7-diamine :
Compound tert-butyl 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin- 3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yS(piperidin-4~ylmethyl)carbamate was stirred in a mixture of TFA, dichloromethane, and water, then concentrated to dryness when the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. This material -was then treated with NBS (1.05 equiv) in acetonitrile. When the reaction was deemed complete by HPLC, LC-MS, or TLC analysis, the mixture was concentrated to dryness, purified by prep-HPLC, and freeze-dried to afford compound 6-bromo-3-(6- fluoroquinolin-3-yl)-N5-(piperidin-4-ylmethyl)pyrazolo[1 ,5"a]pyrimidine-5,7-diamine.
Compound, tert-butyl 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin- 3-yl)pyrazoio[1 ,5-a]pyrimidin-5-y!(piperidin-4-ylmethyl)carbamate (1 equiv) was stirred in a mixture of isocyanate (1.1 equiv), and triethylamine (1.5 equiv) in dichloromethane at 0 °C until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction mixture was poured into saturated aqueous ammonium chloride, and then extracted twice with dichloromethane. The combined organics were dried over sodium sulfate, filtered, concentrated, and purified on silica gel to give the desired product. This material was stirred in a mixture of TFA, dichloromethane, and water, and then concentrated to dryness when the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. This material was then treated with NBS (1.05 equiv) in acetonitrile. When the reaction was deemed complete by HPLC, LC- MS, or TLC analysis, the mixture was concentrated to dryness, purified by prep- HPLC, and freeze-dried to afford compound. Synthesis of 4-((7-amino-6-cvano-3-(6-fiuoroquinoiin-3-yl)pyrazolof1 ,5-alpyrimidin-5- ylamino)methyl)-N-methylpiperidine-1-carboxamide:
Figure imgf000624_0001
SCHEME-48 Compound, tert-butyl 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinoiin- 3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yI(piperidin~4-yimethyl)carbamate (1 equiv) was stirred in a mixture of methyl isocyanate (1.1 equiv), and triethylamine (1.5 equiv) in dichloromethane at 0 °C until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction mixture was poured into saturated aqueous ammonium chloride, and then extracted twice with dichloromethane. The combined organics were dried over sodium sulfate, filtered, concentrated, and purified on silica gel to give the desired product. This material (1 equiv) and NBS (1.05 equiv) in acetonitriie was stirred at room temperature until the reaction was deemed complete by HPLC, LC- MS, or TLC analysis. The reaction mixture was concentrated, re-dissolved in dichloromethane, washed with saturated aqueous sodium thiosulfate, dried over sodium sulfate, filtered, concentrated, and purified on silica gel. The resulting material was stirred with the palladium catalysts (0.2 equiv each) and stannane (1.5 equiv) in dioxane at 160 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The reaction was allowed to cool to room temperature, diluted with ethyl acetate, washed with brine, dried over sodium sulfate, filtered, concentrated, and purified in silica gel. The resulting material was stirred in TFA and water at room temperature until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The mixture was concentrated, purified by prep-HPLC, and freeze-dried to afford compound 4-((7-amino-6-cyano-3-(6-fluoroquinolin-3-yl) pyrazolo [1 , 5-a] pyrimidin-5-yiamino) methyl)-N-methylpiperidine-1 -carboxamide.
Genera! Synthesis of 1-(4-((7-amino-6-bromo-3-(6-fluoroquinolin-3-v1)pyrazolori ,5- aipyrimidin-5-ylamino)alkvQpiperidin-1-yl)ethanone:
Figure imgf000624_0002
SCHEM E-49 Sodium cyanoborohydride ( 3.5 equiv) was added to a stirring mixture of 7-(bis((2- (tπmethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3-yl)pyrazolo[1 ,5-a]pyrimidine-- 5-carbaidehyde (1 equiv) and amine (2 equiv) in ethanol and acetic acid at room temperature. When the reaction was deemed complete by HPLC, LC-MS, or TLC analysis, the mixture was quenched with saturated aqueous sodium bicarbonate, and then extracted with dichloromethane (2x). The combined organics were dried over sodium sulfate, filtered, concentrated to dryness, and purified on siiica gel to afford the desired product, N5-({1-benzylpipeπdin-4-yl)methyl)-3-(6-fluoroquinolin-3-yl)-N7,N7- bis((2-(trimethylsilyl)ethoxy)methyl)pyrazolo[1 ,5-a]pyrimidine-5,7-diamine.
This material, (1 equiv), BoC2O (2 equiv), DMAP (0.2 equiv), and triethylamine (3 equiv) in dichloromethane was stirred at room temperature until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The mixture was diluted with additional dichloromethane, washed with saturated aqueous ammonium chloride, dried over sodium sulfate, filtered, concentrated, and purified on siiica gel to give the product, tert-butyl {1-benzylpiperidin-4-yl)methyl(7-(bis((2-
(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin- 5-yl)carbamate. This material was then stirred with Pd(OH)2/C (0.2 equiv) and ammonium formate (20 equiv) in ethanol at 80 °C until the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. The mixture was allowed to cool to room temperature, filtered through a pad of ceiite, concentrated and purified on silica gel to afford compound tert- butyl 7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl(piperidin-4-ylmethyl)carbamate.
This material was stirred with carboxyiic acid (1.5 equiv), HATU (1.5 equiv), and DIPEA (3 equiv) in DMF until the reaction was deemed complete by TLC, HPLC, or LC-MS analysis. The reaction mixture was concentrated, the resulting residue dissolved in ethyl acetate, washed with water, saturated aqueous sodium bicarbonate, and brine, dried over sodium sulfate, filtered, concentrated, and purified on silica gel to give the product. This material was stirred in a mixture of TFA, dichloromethane, and water, then concentrated to dryness when the reaction was deemed complete by HPLC, LC-MS, or TLC analysis. This material was then treated with NBS (1.05 equiv) in acetonitrile. When the reaction was deemed complete by HPLC, LC-MS, or TLC analysis, the mixture was concentrated to dryness, purified by prep-HPLC, and freeze- dried to afford compound 1 -(4-((7-amino-6-bromo-3-(6-fluoroquinoiin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-ylamino)methyl)piperidin-1 -yi)ethanone.
Synthesis of 1-(4-((7-amino-6-bromo-3-(6-fluoroαuinolin-3-yl)pyrazoioM ,5-aipyrimidin- 5-yl)methylamino)piperidin-1 -yl)-2-methoxypropan-1 -one:
Scheme-49
Figure imgf000626_0001
To a solution of tert-butyl (7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6- f!uoroquinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)methyl(piperidin-4-yl)carbamate (190 mg, 0.25 mmoi) in dichloromethane (3 mL) was added imidazolium salt (84 mg, 0.25mmol) {prepared according to the literature procedure:Grzyb, J.A. et. al. Tetrahedron, 2005, 61, 7153-7175) and Et3N (35 μl_, 0.25 mmol). The resulting mixture was stirred at 25 °C for 16 hours to give. The reaction mixture was diluted with CH2CI2 (50 mL) and was washed with 0.1 N HCI (3 mL), brine (10 mL), dried over Na2SO4. LCMS indicated that the crude product, crude tert-butyl (7-(bis((2- {trimethylsiIyl)ethoxy)methyl)amino)-3-(6-fluoroquinolin-3-yl)pyrazolo[1 ,5-a]pyrimidin- 5-yl)methyl(1-(2-methoxypropanoyl)piperidin-4-yl)carbamate was clean, This material was then converted to the final compound, by bromination with NBS and followed by treatment with 4NHCI, to give 1-(4-((7-amino-6-bromo-3-(6-fluoroquinotin-3- yl)pyrazolo[1 ,5-a]pyrimidin-5-yl)methy!amino)piperidin-1-yl)-2-methoxypropan-1 -one following a similar procedure as that described above schemes: MW: 556.13; MS MH+ m/z: 557; HPLC tR: 4.17. . LCMS: 2.67 mins, m/z =485.0 (MH+).
By essentially the same procedure given in Schemes 45 through 49, the compounds listed ϊn Table 14 can be prepared
TABLE-14
Figure imgf000626_0002
Figure imgf000627_0001
Figure imgf000628_0001
Figure imgf000629_0001
Figure imgf000630_0001
Figure imgf000631_0001
Figure imgf000632_0001
Figure imgf000633_0001
Figure imgf000634_0001
Figure imgf000635_0001
Figure imgf000636_0001
Figure imgf000637_0001
Figure imgf000638_0001
Figure imgf000639_0001
Figure imgf000640_0001
Figure imgf000641_0003
Figure imgf000641_0002
6-(7-amino<3-(quinoiin-3-vl)pvrazolof1 ,5-aipvrimidin-5-vl)hexanoic acid
Figure imgf000641_0001
Scheme 70
To a solution of methyl hex-5-enoate (12 mg, 0.1 mmol) in THF (0.5 m!) under argon at 0 °C was added 9-BBN (0.2 mmol, 0.4 mi of 0.5 M solution in THF). The mixture was warmed to room temperature and stirred for 16 h. Potassium phosphate (3 M in H2O, 0.2 mmol) was added followed by the addition of 5-chloro-3-(quino!in-3-yl)-N,N- bis((2-(trimethylsilyi)ethoxy)methyl) pyrazolo[1 ,5-a]pyπmidin-7-amine (50 mg, 0.09 mmol) in DMF (0.5 ml) and Pd(dppf)Cl2-CH2Ci2 (5 mg, 0.005 mmol). The reaction mixture was heated at 90 °C for 16 h under argon. The solution was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 2:1 THF: MeOH (3 ml_) and was treated with 2N NaOH<aq) (0.25 mL). The resulting solution was stirred at room temperature for 4 hours followed by the addition of 2N hydrochloride solution (1.0 mi). The solution was heated at 65 °C for 2 h, cooled to room temperature, concentrated in vacuo and purified by prep-LC to afford the title compound (8.2 mg): LC/MS RT = 2.74 min. Mass calculated for, M+H 376.17, observed 376.17.
By essentially the same procedure given in Scheme 50, the compounds listed in Table 15 can be prepared.
Table 15
Figure imgf000642_0001
Incorporation of deuterium at the 3-position
Figure imgf000643_0001
Scheme 51
Synthesis of Methyl 4-(7-(bis((2-(trimethylsiM)ethoxy)methyDamino)-3-(6- chloropyridin-3-yl)pyrazoloH ,5-alpyrimidin-5-yl)cvc!ohexanecarboxy[ate
Figure imgf000643_0002
2-Chioro-5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)pyridine (0.55 mmol, 131 rng), K3PO4 (1.36 mmol, 290 mg), and PdCI2{dppf) ■ CH2CI2 (0.045 mmol, 37 mg) is added to a solution of 4-{7-[Bis-(2-trimethylsilanyl-ethoxymethy!)-amino]-3-iodo-pyrazolo[1 ,5- a3pyrimidin-5-yl}-cyciohexanecarboxylic acid methyl ester (0.45 mmol, 300 mg) in dioxane (3.8 ml_). To this suspension is added distilled H2O (0.38 mL). The resulting solution is stirred at 90° C under argon for 18 hours. The reaction mixture is concentrated in vacuo and then purified via silica gel chromatography (0% to 60% ethyl acetate in hexanes gradient) to yield the title compound (220 mg, 75% yield) as yellow oil. LC-MS: 646 [M+H]. Synthesis of (1 r.4r)-methyl 4-(7-(bis((2-(trimethylsilyl)ethoxy)methyQamino)-3-(6- phenyl-d5-pyridin-3-yl)pyrazoloπ ^-alpyrimidin-5-yl)cvclohexanecarboxylate
Figure imgf000644_0001
Phenyl-d5-boronic acid (1.08 mmol, 137 mg), K3PO4 (1.63 mmol, 345 mg), and PdCI2(dppf) • CH2CI2 (0.054 mmol, 45 mg) is added to a solution of methyl 4-(7- (bis((2-(trimethylsilyl)ethoxy)methyl)amino)-3-(6-chloropyridin-3-yi)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexanecarboxylate (0.54 mmol, 350 mg) in dioxane (6.0 ml_). To this suspension is added distilled H2O (0.6 mL). The resulting solution is stirred at 100° C under argon for 18 hours. The reaction mixture is concentrated in vacuo and then purified via silica gel chromatography (0% to 60% ethyl acetate in hexanes gradient) to yield the title compound.
Synthesis of (1r,4r)-4-(7-amino-6-bromo-3-(6-phenyl-d5-pyridin-3-vQpyrazolori ,5- alpyrSmidin-5-yl)cvclohexanecarboxylic acid
Figure imgf000644_0002
N-bromosuccinimide (73 mg, 0.41 mmol) is added to a solution of <1 r,4r)-methyl 4-(7- (bis((2-(trimethyls!lyl)ethoxy)methyl)amino)-3-(6-phenyl-d5"pyπdin-3-yl)pyrazolo[1 ,5- a]pyrimidin-5-yl)cyclohexanecarboxyiate (284 mg, 0.41 mmoi) in acetonitrile (4 mL). The resulting solution is stirred at room temperature for 1 hour. The reaction mixture is concentrated in vacuo and dissolved in 3:3:2 MeOH:THF:H2O (8 mL), followed by the treatment with 3.75 N NaOH(aq) (0.6 mL). The resulting solution is stirred at room temperature for 18 hours. The reaction mixture is treated with 1 N hydrochloride solution (4 ml) at 65 °C for 4 h. The reaction solution is concentrated and purified by prep-LC to afford the title compound.
incorporation of deuterium at the 5-position
Figure imgf000645_0001
Scheme 52
Synthesis of ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)methyl)amino)pyrazoloπ ,5- alpyrimidin-5-yl)cvclohexyl)acetate
Figure imgf000645_0002
To a 50 mL roundbottom flask is charged ethyl 2-(4-(7-(bis((2-(trimethylsilyl)ethoxy)- methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5-yl)cyclohexylidene)acetate (800 mg, 1.43 mmol) and ethyl acetate (15 mL). The flask is flushed with argon and 5% palladium on carbon (100 mg) is added. The flask is sealed and degassed under vacuum. Deuterium gas is then added via balloon. The reaction is stirred under D2 atmosphere 18 hours. The reaction is then filtered through celite to yield deuterated ethyl 2-{4-(7- (bis((2-(trimethylsily!)ethoxy)methyl)amino)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate (761 mg, 1.35 mmol, 95% yield) as pale yellow oil.
Incorporate deuterium at both 3- and 5-positions
Figure imgf000646_0001
Scheme 53 Synthesis of ethyl 2-(4-(7-amino-6-bromo-3-(6-phenylpyridin-3-yl)pyrazolori ,5- aipyrimidin-5-yl)cvclohexyl)acetate
Figure imgf000646_0002
By essentialiy the same procedure given in Schemes 16, 17 and 18, ethyl 2-(4-(7- amino-6-bromo-3-(6-phenyIpyr!din-3-yl)pyrazolo[1 ,5-a]pyrimidin-5- yl)cyclohexyl)acetate can be prepared.
ASSAYS: mTOR Kinase Assay
The nπTOR assay buffer contains 10 mM hepes (pH 7.4), 50 mM NaCI, 100 μg/ml BSA, 50 mM B-glycerophosphate, 10 mM MnCI2 and 0.5 mM DTT. 20 ng of mTOR enzyme is preincubated with the compound for 10 minutes. 5 μM ATP and 0.1 μM GSTS6K is added. The reaction is incubated for one hour at 30 °C. Anti phospho P70S6K (about 1.7 ng/well) and anti GSTXL665 (1 :1 Ratio with the substrate GSTS6K) are added after incubating. The plates are read at least 2 hours after adding the anti phospho p70S6K and the anti GSTXL665. ICm DETERMINATIONS: Dose-response curves were plotted from inhibition data generated, each in duplicate, from 8 point serial dilutions of inhibitory compounds. Concentration of compound was plotted against % kinase activity, calculated by CPM of treated samples divided by CPM of untreated samples. To generate IC5O values, the dose-response curves were then fitted to a standard sigmoidal curve and IC50 values were derived by nonlinear regression analysis.
CHK1 in yltro Kinase Assay
This in vitro assay utilizes recombinant His-CHK1 expressed in the baculovirus expression system as an enzyme source and a biotinyiated peptide based on CDC25C as substrate (biotin-RSGLYRSPSMPENLNRPR). Materials and Reagents:
1 ) CDC25C Ser 216 C-term Biotinyiated peptide substrate (25 mg), stored at -20° C1 Custom Synthesis by Research Genetics: biotin-RSGLYRSPSMPENLNRPR 2595.4 MW 2) His-CHK1 In House lot P976, 235 μg/mL, stored at -80° C.
3) D-PBS (without CaCI and MgCI): GIBCO, Cat.# 14190-144
4) SPA beads: Amersham, Cat.# SPQ0032: 500 mg/vial
Add 10 mL of D-PBS to 500 mg of SPA beads to make a working concentration of 50 mg/mL. Store at 4 °C. Use within 2 week after hydration. 5) 96-WeII White Micropiate with Bonded GF/B filter: Packard, Cat.# 6005177 6) Top seai-A 96 well Adhesive Film: Perkin Elmer, Cat.# 6005185
7) 96-weil Non-Binding White Polystyrene Plate: Corning, Cat. # 6005177
8) MgCl2: Sigma, Cat.# M-8266
9) DTT: Promega, Cat.# V3155 10) ATP, stored at 4 ^C: Sigma, Cat.# A-5394
11 ) γ33p-ATP, 1000-3000 Ci/mMol: Amersham, Cat.# AH9968 12} NaCI: Fisher Scientific, Cat.# BP358-212
13) H3PO4 85% Fisher, Cat.#A242-500
14) Tris-HCL pH 8.0: Bio-Whittaker, Cat. # 16-015V 15) Staurosporine, 100 μg: CALBIOCHEM, Cat. # 569397
16) Hypure Cell Culture Grade Water, 500 mL: HyClone, Cat.# SH30529.02 Reaction Mixtures:
1) Kinase Buffer: 50 mM Tris pH 8.0; 10 mM MgCl2; 1 mM DTT
2) His-CHK1 , In House Lot P976, MW -3OKDa1 stored at -80° C. 6 nM is required to yield positive controls of -5,000 CPM. For 1 piate (100 reaction): dilute 8 μi_ of 235 μg/mL (7.83 μM) stock in 2 mL Kinase Buffer. This makes a 31 nM mixture. Add 20 μL/well. This makes a final reaction concentration of 6 nM.
3) CDC25C Biotinylated peptide.
Dilute CDC25C to 1 mg/mL (385 μM) stock and store at -20 °C. For 1 plate (100 reactions): dilute 10 μL of 1 mg/mL peptide stock in 2 mL Kinase Buffer. This gives a 1.925 μM mix. Add 20 μL/reaction. This makes a final reaction concentration of 385 nM.
4) ATP Mix.
For 1 plate (100 reactions): dilute 10 μL of 1 mM ATP (cold) stock and 2 μL fresh P33-ATP (20 μCi) in 5 mL Kinase Buffer. This gives a 2 μM ATP (cold) solution; add 50 μL/well to start the reaction. Final volume is 100 μL/reaction so the final reaction concentrations will be 1 μM ATP (cold) and 0.2 μCi/reaction.
5) Stop Solution:
For 1 plate add: To 10 mL Wash Buffer 2 (2M NaCI 1 % H3PO4) : 1 mL SPA bead slurry (50 mg); Add 100 μL/well
6) Wash buffer 1 : 2 M NaCl 7) Wash buffer 2: 2 M NaCl, 1% H3PO4 Assay Procedure:
Figure imgf000650_0001
* Total reaction volume for assay.** Final reaction volume at termination of reaction (after addition of stop solution).
1) Dilute test compounds to desired concentrations in water/10% DMSO - this will give a final DMSO concentration of 1 % in the reaction. Dispense 10 μL/reaction to appropriate weils. Add 10 μl_ 10% DMSO to positive (CHK1 +CDC25C+ATP) and negative (CHK1+ATP only) control wells. 2) Thaw enzyme on ice -- dilute enzyme to proper concentration in kinase buffer (see
Reaction Mixtures) and dispense 20 μL to each well.
3) Thaw the Biotinylated substrate on ice and dilute in kinase buffer (see Reaction
Mixtures). Add 20 μL/well except to negative control wells. Instead, add 20 μL Kinase
Buffer to these wells. 4) Dilute ATP (cold) and P33-ATP in kinase buffer (see Reaction Mixtures). Add 50 μL/well to start the reaction.
5) Allow the reaction to run for 2 hours at room temperature.
6) Stop reaction by adding 100 μL of the SPA beads/stop solution (see Reaction Mixtures) and leave to incubate for 15 minutes before harvest 7) Place a blank Packard GF/B filter plate into the vacuum filter device (Packard plate harvester) and aspirate 200 mL water through to wet the system. 8) Take out the blank and put in the Packard GF/B filter plate. 9) Aspirate the reaction through the filter plate.
10) Wash: 200 ml_ each wash; 1 X with 2M NaCI; 1 X with 2M NaCi/ 1 % H3PO4
1 1 ) Allow filter plate to dry 15 minutes.
12) Put TopSeal-A adhesive on top of fitter plate. 13) Run filter plate in Top Count
Settings: Data mode: CPM
Radio nuclide: Manual SPA:P33 Scintillator: Liq/plast Energy Range: Low ICgn DETERMINATIONS: Dose-response curves were plotted from inhibition data generated, each in duplicate, from 8 point serial dilutions of inhibitory compounds. Concentration of compound was plotted against % kinase activity, calculated by CPM of treated samples divided by CPM of untreated samples. To generate IC50 values, the dose-response curves were then fitted to a standard sigmoida! curve and IC50 values were derived by nonlinear regression analysis.
Selected Thiazole Derivatives of the present invention were tested using this assay and provided IC50 values ranging from about 1 nM to about 5500 nM.
CDK2 Kinase Assay BACULQVIRUS CONSTRUCTIONS: Cyclin E was cloned into pVL1393
(Pharmingen, La JoIIa, California) by PCR, with the addition of 5 histidine residues at the amino-terminal end to allow purification on nickel resin. The expressed protein was approximately 45kDa. CDK2 was cloned into pVL1393 by PCR, with the addition of a haemaglutinin epitope tag at the carboxy-terminal end (YDVPDYAS). The expressed protein was approximately 34kDa in size.
ENZYME PRODUCTION: Recombinant bacuioviruses expressing cyclin E and CDK2 were co-infected into SF9 celis at an equal multiplicity of infection (MOi=5), for 48 hrs. Cells were harvested by centrifugation at 1000 RPM for 10 minutes, then pellets lysed on ice for 30 minutes in five times the peiiet volume of lysis buffer containing 5OmM Tris pH 8.0, 15OmM NaCl, 1% NP40, 1 mM DTT and protease inhibitors (Roche Diagnostics GmbH, Mannheim, Germany). Lysates were spun down at 15000 RPM for 10 minutes and the supernatant retained. 5mL of nickel beads (for one liter of SF9 cells) were washed three times in lysis buffer (Qiagen GmbH, Germany). Imidazole was added to the baculovirus supernatant to a final concentration of 2OmM, then incubated with the nickel beads for 45 minutes at 4° C. Proteins were eluted with lysis buffer containing 25OmM imidazole. Eluate was dialyzed about 15 hours in 2 liters of kinase buffer containing 5OmM Tris pH 8.0, 1 mM DTT, 1OmM MgCI≥, 100μM sodium orthovanadate and 20% glycerol. Enzyme was stored in aliquots at -7O°C.
In Vitro Cyclin E/CDK2 Kinase Assays Cyclin E/CDK2 kinase assays can be performed as described below in low protein binding 96-we!S plates {Corning Inc, Corning, New York).
Enzyme is diluted to a final concentration of 50 μg/mL in kinase buffer containing 5OmM Tris pH 8.0, 10 mM MgCI2J mM DTT, and 0.1 mM sodium orthovanadate. The substrate used in these reactions is a biotinylated peptide derived from Histone H1 (from Amersham, UK). The substrate is thawed on ice and diluted to 2 μM in kinase buffer. Test compounds are diluted in 10% DMSO to desirable concentrations. For each kinase reaction, 20 μl_ of the 50 μg/mL enzyme solution (1 μg of enzyme) and 20 μl of the 2 μM substrate solution are mixed, then combined with 10 μL of diluted compound in each well for testing. The kinase reaction is initiated by addition of 50 μL of 2 μM ATP and 0.1 μCi of 33P-ATP (from Amersham, UK). The reaction iss allowed to run for 1 hour at room temperature, then is stopped by adding 200 μL of stop buffer containing 0.1 % Triton X-100, 1 mM ATP, 5mM EDTA, and 5 mg/mL streptavidine coated SPA beads (from Amersham, UK) for 15 minutes. The SPA beads are then captured onto a 96-well GF/B filter plate (Packard/Perkin Elmer Life Sciences) using a Filtermate universal harvester (Packard/Perkin Elmer Life Sciences.). Non-specific signals are eliminated by washing the beads twice with 2M NaCI then twice with 2 M NaCI with 1% phosphoric acid. The radioactive signal can then be measured using, for example, a TopCount 96 well liquid scintillation counter (from Packard/Perkin Elmer Life Sciences). ICgi DETERMINATIONS: Dose-response curves are plotted from inhibition data generated, each in duplicate, from 8 point serial dilutions of inhibitory compounds. Concentration of compound is plotted against % kinase activity, calculated by CPM of treated samples divided by CPM of untreated samples. To generate IC50 values, the dose-response curves are then fitted to a standard stgmoidal curve and IC50 values can be derived using nonlinear regression analysis.
Compounds of the present invention exhibit mTOR IC50 values of about 1 nM to about 5500 nM, CHK1 IC50 values of about 100 nM to about 55000 nM, and CDK2 IC50 values of about 800 nM to about 30000 nM. In all cases, the compounds are much more selective for mTOR over CHK1 and CDK2. Table 15 shows the activity data for an illustrative list of compounds of the invention.
Table 15
Figure imgf000653_0001
Figure imgf000654_0002
Table 16 also lists compounds of the invention with activity data whereby the IC50 values are rated "A", "B1" "C," "D," "E" or T." The IC50 values are rated "A" for IC50 values less than 100 nM, "B" for ICs0 values in the range from 101 nM to 200 nM, "C" for IC50 values in the range from 201 nM to 500 nM, "D" for IC50 values in the range of about 501 nM to 1000 nM, Ε" for IC50 values in the range from 1001 nM to 5000 nM and "F" for IC50 values greater than about 5000 nM.
Table 16
Figure imgf000654_0001
Figure imgf000655_0001
Figure imgf000656_0001
Figure imgf000657_0001
Figure imgf000658_0001
Figure imgf000659_0001
Figure imgf000660_0001
Figure imgf000661_0001
Figure imgf000662_0001
Whiie the present invention has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and other variations thereof will be apparent to those of ordinary skill in the art. All such alternatives, modifications and variations are intended to fall within the spirit and scope of the present invention.

Claims

What is claimed is: 1. A compound of the formuia:
Figure imgf000663_0001
Figure imgf000664_0001
Figure imgf000665_0001
Figure imgf000666_0001
Figure imgf000667_0001
Figure imgf000668_0001
668
Figure imgf000669_0001
Figure imgf000670_0001
670
Figure imgf000671_0001
Figure imgf000672_0001
Figure imgf000673_0001
Figure imgf000674_0001
674
Figure imgf000675_0001
Figure imgf000676_0001
Figure imgf000677_0001
Ķ77
Figure imgf000678_0001
Figure imgf000679_0001
Figure imgf000680_0001
Figure imgf000681_0001
Figure imgf000682_0001
Figure imgf000683_0001
Figure imgf000684_0001
Figure imgf000685_0001
Figure imgf000686_0001
Figure imgf000687_0001
Figure imgf000688_0001
Figure imgf000689_0001
Figure imgf000690_0001
Figure imgf000691_0001
Figure imgf000692_0001
Figure imgf000693_0001
Figure imgf000694_0001
Figure imgf000695_0001
Figure imgf000696_0001
Figure imgf000697_0001
Figure imgf000698_0001
Figure imgf000699_0001
Figure imgf000700_0001
Figure imgf000701_0001
Figure imgf000702_0001
Figure imgf000703_0001
Figure imgf000704_0001
Figure imgf000705_0001
Figure imgf000706_0001
Figure imgf000707_0001
Figure imgf000708_0001
Figure imgf000709_0001
Figure imgf000710_0001
Figure imgf000711_0001
Figure imgf000712_0001
Figure imgf000713_0001
Figure imgf000714_0001
Figure imgf000715_0001
Figure imgf000716_0001
or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
2. A pharmaceutical composition comprising a therapeutically effective amount of at least one compound of claim 1 or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, in combination with at least one pharmaceutically acceptable - carrier.
3. The pharmaceutical composition according to claim 2, further comprising one or more anti-cancer agents different from the compounds of claim 1.
4. The pharmaceutical composition according to claim 3, wherein the one or more anti-cancer agents are selected from the group consisting of Adriamycin, Altretamine, Amidox, Aminoglutethimide, Amsacrine, Anastrazole, Antibodies to EGFR, 3-AP, Aphidicolon, Ara-C, Arsenic trioxide, L-Asparaginase, Bevacizumab, Bleomycin, BMS 214662, Bortezomib, Busulfan, Campath, Camptostar, Capecitabine, Carboplatin, Carmustine, Centrosome associated protein E ("CENP-E") inhibitors, Cetuximab, Cladribine, Chlorambucil, Chlormethine, Chlorotrianisene, Cisplatin, Clofarabine, cyclophosphamide, Cytarabine, a Cytostatic agent, Cytoxan, Dacarbazine, Dactinomycin, Daunorubicin, Dasatinib, Deforolimus, Deoxycoformycin, Didox,
Diethylstilbestrol, Docetaxel, Doxorubicin, Dromostanolone, Droloxafine, Epirubicin, Epothilones, ERK inhibitors, Erlotinib, Etoposide, 17α-Ethinylestradiol, Estramustine, Exemestane, Floxuridine, Fludarabine, Fludarabine phosphate, 5-Fluorouracii, Fluoxymesterone, Flutamide, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab ozogamcicin, Gosereliπ, GSK-923295, Hexamethylmelamine, Hydroxyprogesterone, Hydroxyurea, lbritumomab Tiuxetan, [darubicin, Ifosfamide, lmatinib mesylate, Intron, Irinotecan, ispinesib, KSP inhibitors, L778.123, Lapatinib, Leucovirin, Leuprolide, Lerozole, Letrazole, Levamisole, Liposomal Doxorubicin, Liposomal, Lomustine, lonafornib, Medroxyprogesteroneacetate, Megestrolacetate, Melphalan, 6-Mercaptopurine, Methoxtrexate, Methylprednisoione, Methyltestosterone, Mithramycin, Mitomycin-C, Mitotane, Mitoxantrone, Navelbene, Niiotinib, Oxaliplatin, Paclitaxel, Panitubimab, Pentostatin, Pipobroman, Porfimer, Prednisolone, Prednisone propionate, Procarbazine, Reioxafine, Rituximab, Satriplatin, SB-743921, SmII , Sorafinib, Streptozocin, Sunitinib, Tamoxifen, Taxotere, Taxol, Temozolomide, Teniposide, Testolactone, Testosterone, Tezacitabine, 6-Thioguanine, Thiotepa, Tipifarnib, Topotecan, Toremifene, Tositumomab, Trastuzumab, Triamcinolone, Triapine, Triethylenemelamine, Triethylenethiophosphoramine, Trimidox, Uracil mustard, Vinblastine, Vincristine, Vindesine, and Vinoreibine.
5. A method of treating, or slowing the progression of, a disease by inhibiting mammalian Target Of Rapamycin in a patient, said method comprising administering a therapeutically effective amount of at least one compound of claim 1 , or a pharmaceuticaliy acceptable salt, solvate, ester or prodrug of the compound, to a patient in need thereof.
6. A method of treatment of a disease selected from the group consisting of proliferative inflammatory diseases, allergic diseases, obstructive airways diseases, diseases related to transplant rejection and diseases that respond to inhibition of mTOR, comprising administering a therapeutically effective amount of compound of claim 1 or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof to a patient in need of such treatment.
7. A method of treating a disease by inhibiting a mTOR, comprising administering to a patient in need of such treatment an amount of a first compound of claim 1 or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof; and an amount of at least one second compound, said second compound being an anti-cancer agent; wherein the amounts of the first compound and said second compound result in a therapeutic effect.
8. The method according to claim 7, wherein the one or more anti-cancer agents are selected from the group consisting of Adriamycin, Altretamine, Amidox, AminogSutethimide, Amsacrine, Anastrazole, Antibodies to EGFR, 3-AP, Aphidicolon, Ara-C, Arsenic trioxide, L-Asparaginase, Bevacizumab, Bleomycin, BMS 214662, Bortezomib, Busuifan, Campath, Camptostar, Capecitabine, Carboplatin, Carmustine, Centrosome associated protein E ("CENP-E") inhibitors, Cetuximab, Cladribine, Chlorambucil, Chiormethine, Chlorotrianisene, Cisplatin, Clofarabine, cyclophosphamide, Cytarabine, a Cytostatic agent, Cytoxan, Dacarbazine, Dactinomycin, Daunorubicin, Dasatinib, Deforolimus, Deoxycoformycin, Didox, Diethylstilbestroi, Docetaxel, Doxorubicin, Dromostanolone, Droloxafine, Epirubicin, Epothilones, ERK inhibitors, Erlotinib, Etoposide, 17α-Ethiny!estradiol, Estramustine, Exemestane, Fioxuridine, Fludarabine, Fludarabine phosphate, 5-F!uorouracil, Fluoxymesterone, Flutamide, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab ozogamcicin, Goserelin, GSK-923295, Hexamethylmelamine, Hydroxyprogesterone, Hydroxyurea, lbritumomab Tiuxetan, Idarubicin, Ifosfamide, lmatinib mesylate, Intron, Irinotecan, ispinesib, KSP inhibitors, L778,123, Lapatinib, Leucovirin, Leuprolide, Lerozole, Letrazole, Levamisole, Liposomal Doxorubicin, Liposomal, Lomustine, Lonafarnib, Medroxyprogesteroneacetate, Megestrolacetate, Melphalan, 6-Mercaptopurine, Methoxtrexate, Methylprednisolone, Methyltestosterone, Mithramycin, Mitomycin-C, Mitotane, Mitoxantrone, Navelbene, Nilotinib, Oxaliplatin, Paclitaxel, Panitubimab, Pentostatin, Pipobroman, Porfimer, Prednisolone, Prednisone propionate, Procarbazine, Reloxafine, Rituximab, Satriplatin, SB-743921 , SmH , Sorafinib, Streptozocin, Sunitinib, Tamoxifen, Taxotere, Taxol, Temozoiomide, Teniposide, Testolactone, Testosterone, Tezacitabine, 6-Thioguanine, Thiotepa, Tipifarnib, Topotecan, Toremifene, Tositumomab, Trastuzumab, Triamcinolone, Triapine, Triethylenemeiamine, Triethylenethiophosphoramine, Trimidox, Uracil mustard, Vinblastine, Vincristine, Vindesine, and Vinorelbine.
9. A method of treating a cancer comprising administering a therapeutically effective amount of at least one compound of claim 1.
10. The method of claim 9, wherein said cancer is selected from the group consisting of: cancer of the bladder, breast, colon, kidney, liver, lung, small cell lung cancer, non-small ceil lung cancer, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, including squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma and Burkett's lymphoma; acute and chronic myelogenous leukemia, myelodysplastic syndrome and promyelocyte leukemia; fibrosarcoma, rhabdomyosarcoma; mantle ceil lymphoma, myeloma; astrocytoma, neuroblastoma, glioma and schwannomas; melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderma pigmentosum, keratoctanthoma, thyroid follicular cancer, endometrial cancer, gastrointestinal tract cancer and Kaposi's sarcoma.
1 1. A method of inhibiting a mammalian Target Of Rapamycin, comprising administering a therapeutically effective amount of a compound of claim 1.
12. A method of treating a disease by inhibiting a mTOR, comprising administering to a patient in need of such treatment an amount of a first compound represented by the structural Formula I:
Figure imgf000719_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein:
R is independently selected from the group consisting of halo, hydroxyl, amino, -CN, H, -(CrC6)alkyl, alkoxy, -C(=O)alkyi, heteroaryl and aryl, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl or halo; R1 is independently selected from the group consisting of heterocycloalkyl, heterocycioalkylalkyl, spiroheterocycloalkyl, heterocyclenyl, -NR3R4, cycloaikyl, heteroaryl, aryl, alkynyi, heterocyclenylalkyi, cycloalkylalkyl, heteroarylaikyl heteroarylalkynyl, spiroheterocycloalkylalkyl, -N-heteroaryl, and arylalkyl, wherein each of said heterocycloalkyl, heterocycloalkyfaJkyl, spiroheterocycloaikyl, heterocycienyl, cycloaikyl, heteroaryl, aryl, alkynyi, heterocycienylalky!, cycloalkylaikyl, heteroarylaikyl, heteroarylalkynyl , -N-heteroaryi and arylalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloaikyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -a]kyl(CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2l -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyl, halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylaikyl, -C(O)-heteroaryl, -C(O)-alkyl-O-alkyl, - aikyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S(O)2-CF3, -N-aikyl, -SOa-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2l -alkyl-NS(O)2-alkyl, alkyl(CO)NS(O)2- cycloalkyl, -CO-CO2H1 -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C(O)-heteroaryl can be unsubstituted or substituted with one or more alkyl; R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3)2CN, trifiuromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl, and arylalkyl; R3 is cycloaikyl or heteroaryl, wherein each of said cycloaikyl or heteroaryl can be unsubstituted or substituted with one or more moieties independently selected from the group consisting of X; and
R4 is H; and an amount of at least one second compound, said second compound being an anti-cancer agent; wherein the amounts of the first compound and said second compound result in a therapeutic effect.
13. The method according to claim 12, wherein the one or more anti-cancer agents are selected from the group consisting of Adriamycin, Altretamine, Amidox, Aminoglutethimide, Amsacrine, Anastrazole, Antibodies to EGFR, 3-AP, Aphidicolon, Ara-C, Arsenic trioxide, L-Asparaginase, Bevacizumab, Bleomycin, BMS 214662, Bortezomib, Busulfan, Campath, Camptostar, Capecitabine, Carboplatin, Carmustine, Centrosome associated protein E ("CENP-E") inhibitors, Cetuximab, Cladribine, Chlorambucil, Chlormethine, Chlorotrianisene, Cisplatin, C10farabine, cyclophosphamide, Cytarabine, a Cytostatic agent, Cytoxan, Dacarbazine, Dactinomycin, Daunorubicin, Dasatinib, Deforolimus, Deoxycoformycin, Didox, Diethylstilbestrol, Docetaxel, Doxorubicin, Dromostanoione, Droloxafine, Epirubicin, Epothilone, ERK inhibitors, Erlotinib, Etoposide, 17tx-Ethinylestradiol, Estramustine, Exemestane, Fioxuridine, Fludarabine, Fludarabine phosphate, 5-Fluorouracil, Fluoxymesterone, Flutamide, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab ozogamcicin, Goserelin, GSK-923295, Hexamethylmelamine, Hydroxyprogesterone, Hydroxyurea, lbritumomab Tiuxetan, idarubicin, Ifosfamide, lmatinib mesylate, Intron, Irinotecan, ispinesib, KSP inhibitors, L778,123, Lapatinib, Leucovirin, Leuprolide, Lerozoie, Letrazole, Levamisole, Liposomal Doxorubicin, Liposomal, Lomustine, Lonafarnib, Medroxyprogesteroneacetate, Megestrolacetate, Melphatan, 6-Mercaptopurine, Methoxtrexate, Methylprednisolone, Methyltestosterone, Mithramycin, Mitomycin-C, Mitotane, Mitoxantrone, Navelbene, Nilotinib, Oxaliplatin, Paclitaxel, Panitubimab, Pentostatin, Pipobroman, Porfimer, Prednisolone, Prednisone propionate, Procarbazine, Reloxafine, Rituximab, Satriplatin, SB-743921 , SmM , Sorafinib, Streptozocin, Sunitinib, Tamoxifen, Taxotere, Taxol, Temozolomide, Teniposide, Testolactone, Testosterone, Tezacitabine, 6-Thioguanine, Thiotepa, Tipifarnib, Topotecan, Toremifene, Tositumomab, Trastuzumab, Triamcinolone, Triapine, Triethylenemelamine, Triethylenethiophosphoramine, Trimidox, Uracil mustard, Vinblastine, Vincristine, Vindesine, and Vinorelbine.
14. A method of treating a cancer comprising administering a therapeutically effective amount of at least one compound represented by the structural Formula I:
Figure imgf000722_0001
Formula I or a pharmaceutically acceptable salt, solvate, ester, or prodrug thereof, wherein: R is independently selected from the group consisting of halo, hydroxyl, amino,
-CN, H, -(CrCβ)alkyΙ, alkoxy, -C(=O)a!kyl, heteroaryl and aryi, wherein each of said heteroaryl and aryl can be unsubstituted or substituted with one or more alkyl or halo;
R1 is independently selected from the group consisting of heterocycloalkyl, heterocycloalkylalkyl, spiroheterocycloalkyl, heterocyclenyl, -NR3R4, cycloalky!, heteroaryl, aryl, alkynyl, heterocyclenylaikyl, cycloalky I alkyl, heteroarylalky^ heteroarylalkynyl, spiroheterocycloalkylalkyi, -N-heteroaryl, and arylalkyl, wherein each of said heterocycloalkyl, heterocycloalkylalkyl, spiroheterocycloalkyl, heterocyclenyl, cycioaikyl, heteroaryl, aryl, alkynyl, heterocyclenylaikyl , cycloalkyialkyl, heteroarylalkyl, heteroaryialkynyl, -N-heteroaryl and arylalkyl can be unsubstituted or substituted with one or moieties independently selected from the group X;
X is alkoxyl, alkyl, -C(O)alkyl, -C(O)-hydroxyalkyl, -C(O)2alkyl, -C(O)2H, hydroxyalkyl, -S(O)2alkyl, hydroxyl, heterocycloalkyl, -NH-heterocycloalkyl, - trihaloalkyl, -dihaloalkyl, -monohaloalkyl, -N-S(O)2-alkyl, -C(O)-heteroaryl, -alkyl- C(O)2H, -alkyl{CO)N(CH3)-O-CH3, -alkyl(CO)-heteroaryl, -C(O)2-alkyl, -alkyl-C(O)-NH2j -NH2, heteroaryl, -alkyl-CN, -C(O)2-arylalkyi( halo, carboxyesteralkyl, -C(O)-NH2, - alkyl-C(O)2alkyl, heteroarylalkyl, -C(O)-heteroaryl, -C(O)-alkyi-O-alkyl, - alkyl(CO)NS(O)2-cycloalkyl, -alkyl(CO)N-S{O)2-CF3, -N-alkyl. -SO2-cycloalkyl, - alkyl(CO)NS(O)2-alkyl, -alkyl-C(O)-N(alkyl)2, -alkyl-NS<O)2-alkyl, alky!(CO)NS(O)2- cycloalkyl, -CO-CO2H, -C(O)2-alkyl-aryl, -SO2-CF3 or -C(O)H, wherein each of said heterocycloalkyl, heteroaryl or -C (O)- heteroaryl can be unsubstituted or substituted with one or more alkyl;
R2 is heteroaryl or aryl, wherein each of said heteroaryl or aryl can be unsubstituted or independently substituted with one or more moieties independently selected from the group consisting of alkyl, alkoxyl, -CN, aryloxyl, aryl, halo, hydroxyl, -C(CH3^CN, trifluromethyl, difluromethyl, monofluromethyl,_heterocycloalkyl and arylalkyl;
R3 is cycloalkyl or heteroaryi, wherein each of said cycloalkyl or heteroaryl can be unsubstituted or substituted with one or more moieties independently selected from the group consisting of X; and R4 is H.
15. The method of claim 14, wherein said cancer is selected from the group consisting of: cancer of the bladder, breast, colon, kidney, liver, lung, small cell lung cancer, non-smail cell lung cancer, head and neck, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, including squamous cell carcinoma; leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, mantle cell lymphoma, myeloma and Burkett's lymphoma; acute and chronic myelogenous leukemia, myeiodysplastic syndrome and promyelocytic leukemia; fibrosarcoma, rhabdomyosarcoma; mantle cell lymphoma, myeloma; astrocytoma, neuroblastoma, glioma and schwannomas; melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderma pigmentosum, keratoctanthoma, thyroid follicular cancer, endometrial cancer, gastrointestinal tract cancer and Kaposi's sarcoma.
PCT/US2010/030350 2009-04-09 2010-04-08 Pyrazolo [1, 5-a] pyrimidine derivatives as mtor inhibitors WO2010118207A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP10715010.4A EP2417138B1 (en) 2009-04-09 2010-04-08 Pyrazolo[1, 5-a]pyrimidine derivatives as mtor inhibitors
US13/263,193 US8591943B2 (en) 2009-04-09 2010-04-08 Pyrazolo[1,5-a]pyrimidine derivatives as mTOR inhibitors

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US16809309P 2009-04-09 2009-04-09
US61/168,093 2009-04-09

Publications (1)

Publication Number Publication Date
WO2010118207A1 true WO2010118207A1 (en) 2010-10-14

Family

ID=42936570

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2010/030350 WO2010118207A1 (en) 2009-04-09 2010-04-08 Pyrazolo [1, 5-a] pyrimidine derivatives as mtor inhibitors

Country Status (4)

Country Link
US (1) US8591943B2 (en)
EP (1) EP2417138B1 (en)
TW (1) TW201102392A (en)
WO (1) WO2010118207A1 (en)

Cited By (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013016164A1 (en) * 2011-07-26 2013-01-31 Merck Sharp & Dohme Corp. FUSED TRICYCLIC COMPOUNDS AS mTOR INHIBITORS
US20130150362A1 (en) * 2010-08-23 2013-06-13 Merck Sharp & Dohme Corp. NOVEL PYRAZOLO[1,5-a]PYRIMIDINE DERIVATIVES AS mTOR INHIBITORS
WO2013184876A1 (en) 2012-06-06 2013-12-12 Constellation Pharmaceuticals, Inc. Benzo [c] isoxazoloazepine bromodomain inhibitors and uses thereof
US8871754B2 (en) 2012-11-19 2014-10-28 Irm Llc Compounds and compositions for the treatment of parasitic diseases
US8940742B2 (en) 2012-04-10 2015-01-27 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
WO2015037716A1 (en) 2013-09-12 2015-03-19 住友化学株式会社 Nitrogen-containing saturated heterocyclic compound
US9359365B2 (en) 2013-10-04 2016-06-07 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
WO2016142508A1 (en) * 2015-03-11 2016-09-15 Centre Léon-Bérard Composition for treating pancreatic neuroendocrine tumours
US9556169B2 (en) 2012-11-19 2017-01-31 Novartis Ag Compounds and compositions for the treatment of parasitic diseases
JP2017507980A (en) * 2014-03-17 2017-03-23 レクシコン ファーマシューティカルズ インコーポレイテッド Inhibitor of adapter-related kinase 1, composition containing the same, and method of use thereof
US9708348B2 (en) 2014-10-03 2017-07-18 Infinity Pharmaceuticals, Inc. Trisubstituted bicyclic heterocyclic compounds with kinase activities and uses thereof
US9751888B2 (en) 2013-10-04 2017-09-05 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US9775844B2 (en) 2014-03-19 2017-10-03 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
EP3305786A2 (en) 2018-01-22 2018-04-11 Bayer CropScience Aktiengesellschaft Condensed bicyclic heterocycle derivatives as pesticides
US10160761B2 (en) 2015-09-14 2018-12-25 Infinity Pharmaceuticals, Inc. Solid forms of isoquinolinones, and process of making, composition comprising, and methods of using the same
JP2019502678A (en) * 2015-12-10 2019-01-31 ユーシービー バイオファルマ エスピーアールエル Hexahydropyrazinotriazinone derivatives as kinase inhibitors
WO2019054427A1 (en) 2017-09-14 2019-03-21 第一三共株式会社 Compound having cyclic structure
US20200206188A1 (en) * 2012-03-23 2020-07-02 Dennis M. Brown Compositions and methods to improve the therapeutic benefit of indirubin and analogs thereof, including meisoindigo
US10759806B2 (en) 2016-03-17 2020-09-01 Infinity Pharmaceuticals, Inc. Isotopologues of isoquinolinone and quinazolinone compounds and uses thereof as PI3K kinase inhibitors
CN112010860A (en) * 2020-08-05 2020-12-01 南京纳丁菲医药科技有限公司 Benzyloxypyrazolopyrimidine compounds, pharmaceutical compositions and uses thereof
US10919914B2 (en) 2016-06-08 2021-02-16 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
JP2021506954A (en) * 2017-12-15 2021-02-22 ピラミッド バイオサイエンシズ インコーポレイテッド 5- (2- (2,5-difluorophenyl) pyrrolidine-1-yl) -3- (1H-pyrazole-1-yl) pyrazolo [1,5] as a TRK kinase inhibitor for the treatment of cancer -A] Pyrimidine derivatives and related compounds
WO2021122745A1 (en) 2019-12-16 2021-06-24 Carrick Therapeutics Limited 4-[[(7-aminopyrazolo[1,5-a]pyrimidin-5-yl)amino]methyl]piperidin-3-ol compounds and their therapeutic use
WO2021216454A1 (en) * 2020-04-21 2021-10-28 Lexicon Pharmaceuticals, Inc. 4-(3-(pyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5-yl)piperazine for use in the the treatment of cov-229e or cov-oc43 coronaviruses infections
CN114423756A (en) * 2019-06-28 2022-04-29 上海瑛派药业有限公司 Substituted fused heteroaromatic bicyclic compounds as kinase inhibitors and uses thereof
WO2023134692A1 (en) * 2022-01-13 2023-07-20 浙江同源康医药股份有限公司 Polycyclic compound and use thereof

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011090935A1 (en) 2010-01-19 2011-07-28 Merck Sharp & Dohme Corp. PYRAZOLO[1,5-a]PYRIMIDINE COMPOUNDS AS mTOR INHIBITORS
CA2898294C (en) * 2013-02-21 2020-06-09 Calitor Sciences, Llc Heteroaromatic compounds as pi3 kinase modulators
US9944648B2 (en) 2014-01-09 2018-04-17 Intra-Cellular Therapies, Inc. Organic compounds
TWI820146B (en) * 2018-06-15 2023-11-01 瑞典商阿斯特捷利康公司 Purinone compounds and their use in treating cancer
CA3111980A1 (en) 2018-09-10 2020-03-19 Mirati Therapeutics, Inc. Combination therapies
CA3134814A1 (en) 2019-03-26 2020-10-01 Ventyx Biosciences, Inc. Tyk2 pseudokinase ligands
AU2020378345A1 (en) 2019-11-08 2022-06-02 Ventyx Biosciences, Inc. TYK2 pseudokinase ligands
CN113372531B (en) * 2021-05-13 2022-11-25 眉山尤博瑞新材料有限公司 Polyurethane with increased modulus after water absorption and preparation method and application thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006090169A1 (en) * 2005-02-25 2006-08-31 Kudos Pharmaceuticals Limited 2,4-diamino-pyridopyrimidine derivatives and their use as mtor inhibitors
US20070082900A1 (en) * 2005-10-06 2007-04-12 Schering Corporation Methods for inhibiting protein kinases
WO2007044449A2 (en) * 2005-10-06 2007-04-19 Schering Corporation Novel pyrazolopyrimidines as cyclin dependent kinase inhibitors
WO2007061737A2 (en) * 2005-11-17 2007-05-31 Osi Pharmaceuticals, Inc. FUSED BICYCLIC mTOR INHIBITORS
WO2007087395A2 (en) * 2006-01-25 2007-08-02 Osi Pharmaceuticals, Inc. UNSATURATED mTOR INHIBITORS

Family Cites Families (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
HU229024B1 (en) 1996-07-24 2013-07-29 Bristol Myers Squibb Pharma Co Azolo-pyridimidines, pharmaceutical compositions containing the same and use thereof
US20020041880A1 (en) 2000-07-05 2002-04-11 Defeo-Jones Deborah Method of treating cancer
WO2003091256A1 (en) 2002-04-23 2003-11-06 Shionogi & Co., Ltd. PYRAZOLO[1,5-a]PYRIMIDINE DERIVATIVE AND NAD(P)H OXIDASE INHIBITOR CONTAINING THE SAME
EP1534710B1 (en) 2002-09-04 2007-10-24 Schering Corporation Pyrazolopyrimidines as cyclin dependent kinase inhibitors
US7119200B2 (en) 2002-09-04 2006-10-10 Schering Corporation Pyrazolopyrimidines as cyclin dependent kinase inhibitors
EP1537116B1 (en) 2002-09-04 2010-06-02 Schering Corporation Pyrazolopyrimidines suitable for the treatment of cancer diseases
CA2527136A1 (en) 2003-06-03 2004-12-09 Basf Aktiengesellschaft Substituted pyrazolopyrimidines, methods for the production thereof, use of the same for controlling pathogenic fungi, and agents containing said compounds
US20050222171A1 (en) 2004-01-22 2005-10-06 Guido Bold Organic compounds
PE20051089A1 (en) 2004-01-22 2006-01-25 Novartis Ag PYRAZOLE [1,5-A] PYRIMIDIN-7-IL-AMINE DERIVATIVES AS PROTEIN KINASE INHIBITORS
GB0515026D0 (en) 2005-07-21 2005-08-31 Novartis Ag Organic compounds
EP1940839B1 (en) 2005-10-07 2013-07-31 Exelixis, Inc. PYRIDOPYRIMIDINONE INHIBITORS OF PI3Kalpha
GB0525080D0 (en) 2005-12-09 2006-01-18 Astrazeneca Ab Pyrimidine derivatives
CA2657200A1 (en) 2006-07-28 2008-01-31 Novartis Ag 2,4-substituted quinazolines as lipid kinase inhibitors
WO2011002887A1 (en) * 2009-07-02 2011-01-06 Schering Corporation FUSED TRICYCLIC COMPOUNDS AS NOVEL mTOR INHIBITORS

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006090169A1 (en) * 2005-02-25 2006-08-31 Kudos Pharmaceuticals Limited 2,4-diamino-pyridopyrimidine derivatives and their use as mtor inhibitors
US20070082900A1 (en) * 2005-10-06 2007-04-12 Schering Corporation Methods for inhibiting protein kinases
WO2007044449A2 (en) * 2005-10-06 2007-04-19 Schering Corporation Novel pyrazolopyrimidines as cyclin dependent kinase inhibitors
WO2007061737A2 (en) * 2005-11-17 2007-05-31 Osi Pharmaceuticals, Inc. FUSED BICYCLIC mTOR INHIBITORS
WO2007087395A2 (en) * 2006-01-25 2007-08-02 Osi Pharmaceuticals, Inc. UNSATURATED mTOR INHIBITORS

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
A. L. BINGHAM ET AL., CHEM. COMMUN., 2001, pages 603 - 604
CANCER RESEARCH, vol. 57, 1997, pages 3375
J. CELL SCI., vol. 108, 1995, pages 2897
M. CAIRA ET AL., J. PHARMACEUTICAL SCI., vol. 93, no. 3, 2004, pages 601 - 611
P. GOULD, INTERNATIONAL J. OF PHARMACEUTICS, vol. 33, 1986, pages 201 - 217
S. BERGE ET AL., JOURNAL OF PHARMACEUTICAL SCIENCES, vol. 66, no. 1, 1977, pages 1 - 19
T. HIGUCHI; V. STELLA: "Pro-drugs as Novel Delivery Systems", A.C.S. SYMPOSIUM SERIES, 1987
T. HIGUCHI; W. STELLA: "Pro-drugs as Novel Delivery Systems", A.C.S. SYMPOSIUM SERIES, vol. 14
TONDER ET AL., AAPS PHARMSCITECH., vol. 12, 2004

Cited By (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130150362A1 (en) * 2010-08-23 2013-06-13 Merck Sharp & Dohme Corp. NOVEL PYRAZOLO[1,5-a]PYRIMIDINE DERIVATIVES AS mTOR INHIBITORS
EP2608669A1 (en) * 2010-08-23 2013-07-03 Merck Sharp & Dohme Corp. NOVEL PYRAZOLO[1,5-a]PYRIMIDINE DERIVATIVES AS mTOR INHIBITORS
EP2608669A4 (en) * 2010-08-23 2014-01-22 Merck Sharp & Dohme NOVEL PYRAZOLO[1,5-a]PYRIMIDINE DERIVATIVES AS mTOR INHIBITORS
US8883801B2 (en) * 2010-08-23 2014-11-11 Merck Sharp & Dohme Corp. Substituted pyrazolo[1,5-a]pyrimidines as mTOR inhibitors
US8901142B2 (en) 2011-07-26 2014-12-02 Merck Sharp & Dohme Corp. Fused tricyclic compounds as mTOR inhibitors
WO2013016164A1 (en) * 2011-07-26 2013-01-31 Merck Sharp & Dohme Corp. FUSED TRICYCLIC COMPOUNDS AS mTOR INHIBITORS
US20200206188A1 (en) * 2012-03-23 2020-07-02 Dennis M. Brown Compositions and methods to improve the therapeutic benefit of indirubin and analogs thereof, including meisoindigo
US9255108B2 (en) 2012-04-10 2016-02-09 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US8940742B2 (en) 2012-04-10 2015-01-27 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
WO2013184876A1 (en) 2012-06-06 2013-12-12 Constellation Pharmaceuticals, Inc. Benzo [c] isoxazoloazepine bromodomain inhibitors and uses thereof
US8871754B2 (en) 2012-11-19 2014-10-28 Irm Llc Compounds and compositions for the treatment of parasitic diseases
US9556169B2 (en) 2012-11-19 2017-01-31 Novartis Ag Compounds and compositions for the treatment of parasitic diseases
US9926314B2 (en) 2012-11-19 2018-03-27 Novartis Ag Compounds and compositions for the treatment of parasitic diseases
WO2015037716A1 (en) 2013-09-12 2015-03-19 住友化学株式会社 Nitrogen-containing saturated heterocyclic compound
US9359365B2 (en) 2013-10-04 2016-06-07 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US10329299B2 (en) 2013-10-04 2019-06-25 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US9751888B2 (en) 2013-10-04 2017-09-05 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US9828377B2 (en) 2013-10-04 2017-11-28 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
JP2017507980A (en) * 2014-03-17 2017-03-23 レクシコン ファーマシューティカルズ インコーポレイテッド Inhibitor of adapter-related kinase 1, composition containing the same, and method of use thereof
US11541059B2 (en) 2014-03-19 2023-01-03 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US9775844B2 (en) 2014-03-19 2017-10-03 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US10675286B2 (en) 2014-03-19 2020-06-09 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US10941162B2 (en) 2014-10-03 2021-03-09 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US10253047B2 (en) 2014-10-03 2019-04-09 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
US9708348B2 (en) 2014-10-03 2017-07-18 Infinity Pharmaceuticals, Inc. Trisubstituted bicyclic heterocyclic compounds with kinase activities and uses thereof
FR3033499A1 (en) * 2015-03-11 2016-09-16 Centre Leon-Berard COMPOSITION FOR THE TREATMENT OF PANCREATIC NEUROENDOCRINE TUMORS
WO2016142508A1 (en) * 2015-03-11 2016-09-15 Centre Léon-Bérard Composition for treating pancreatic neuroendocrine tumours
US10160761B2 (en) 2015-09-14 2018-12-25 Infinity Pharmaceuticals, Inc. Solid forms of isoquinolinones, and process of making, composition comprising, and methods of using the same
US11939333B2 (en) 2015-09-14 2024-03-26 Infinity Pharmaceuticals, Inc. Solid forms of isoquinolinones, and process of making, composition comprising, and methods of using the same
US11247995B2 (en) 2015-09-14 2022-02-15 Infinity Pharmaceuticals, Inc. Solid forms of isoquinolinones, and process of making, composition comprising, and methods of using the same
JP2019502678A (en) * 2015-12-10 2019-01-31 ユーシービー バイオファルマ エスピーアールエル Hexahydropyrazinotriazinone derivatives as kinase inhibitors
US10759806B2 (en) 2016-03-17 2020-09-01 Infinity Pharmaceuticals, Inc. Isotopologues of isoquinolinone and quinazolinone compounds and uses thereof as PI3K kinase inhibitors
US10919914B2 (en) 2016-06-08 2021-02-16 Infinity Pharmaceuticals, Inc. Heterocyclic compounds and uses thereof
WO2019054427A1 (en) 2017-09-14 2019-03-21 第一三共株式会社 Compound having cyclic structure
JP2021506954A (en) * 2017-12-15 2021-02-22 ピラミッド バイオサイエンシズ インコーポレイテッド 5- (2- (2,5-difluorophenyl) pyrrolidine-1-yl) -3- (1H-pyrazole-1-yl) pyrazolo [1,5] as a TRK kinase inhibitor for the treatment of cancer -A] Pyrimidine derivatives and related compounds
JP7321183B2 (en) 2017-12-15 2023-08-04 ピラミッド バイオサイエンシズ インコーポレイテッド 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-3-(1H-pyrazol-1-yl)pyrazolo[1,5] as TRK kinase inhibitors for treating cancer -A] pyrimidine derivatives and related compounds
EP3305786A2 (en) 2018-01-22 2018-04-11 Bayer CropScience Aktiengesellschaft Condensed bicyclic heterocycle derivatives as pesticides
EP3997087A4 (en) * 2019-06-28 2023-05-31 Impact Therapeutics (Shanghai), Inc Substituted fused heteroaromatic bicyclic compounds as kinase inhibitors and the use thereof
CN114423756A (en) * 2019-06-28 2022-04-29 上海瑛派药业有限公司 Substituted fused heteroaromatic bicyclic compounds as kinase inhibitors and uses thereof
WO2021122745A1 (en) 2019-12-16 2021-06-24 Carrick Therapeutics Limited 4-[[(7-aminopyrazolo[1,5-a]pyrimidin-5-yl)amino]methyl]piperidin-3-ol compounds and their therapeutic use
WO2021216454A1 (en) * 2020-04-21 2021-10-28 Lexicon Pharmaceuticals, Inc. 4-(3-(pyridin-3-yl)pyrazolo[1,5-a]pyrimidin-5-yl)piperazine for use in the the treatment of cov-229e or cov-oc43 coronaviruses infections
CN112010860B (en) * 2020-08-05 2023-03-10 南京纳丁菲医药科技有限公司 Benzyloxypyrazolopyrimidine compounds, pharmaceutical compositions and uses thereof
CN112010860A (en) * 2020-08-05 2020-12-01 南京纳丁菲医药科技有限公司 Benzyloxypyrazolopyrimidine compounds, pharmaceutical compositions and uses thereof
WO2023134692A1 (en) * 2022-01-13 2023-07-20 浙江同源康医药股份有限公司 Polycyclic compound and use thereof

Also Published As

Publication number Publication date
US8591943B2 (en) 2013-11-26
EP2417138B1 (en) 2019-11-27
TW201102392A (en) 2011-01-16
US20120114739A1 (en) 2012-05-10
EP2417138A1 (en) 2012-02-15

Similar Documents

Publication Publication Date Title
EP2417138A1 (en) Pyrazolo [1, 5-a] pyrimidine derivatives as mtor inhibitors
JP7229257B2 (en) GCN2 inhibitors and uses thereof
KR101659193B1 (en) Heteroaryl pyridone and aza-pyridone compounds as inhibitors of btk activity
AU2009276339B2 (en) Pyrimidine compounds, compositions and methods of use
JP6159028B2 (en) Syk inhibitor
JP5152922B2 (en) Use of pyrazolo [1,5-A] pyrimidine derivatives to inhibit protein kinases and methods for inhibiting protein kinases
JP5109109B2 (en) A novel imidazopyrazine as a cyclin-dependent kinase inhibitor
WO2008156614A2 (en) Imidazopyrazines as protein kinase inhibitors
KR20190018645A (en) Positive allosteric modulator of muscarinic acetylcholine receptor M4
CA2628455A1 (en) Imidazopyrazines as protein kinase inhibitors
CA2665127A1 (en) Novel pyrazolopyrimidines as cyclin dependent kinase inhibitors
EP2424859A1 (en) Inhibitors of pi3 kinase and / or mtor
AU2008279776A2 (en) Heterocyclic amide compounds as protein kinase inhibitors
CN113683629B (en) Substituted heteroaryl compounds, compositions and uses thereof
EP2448942B1 (en) FUSED TRICYCLIC COMPOUNDS AS mTOR INHIBITORS

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10715010

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2010715010

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 13263193

Country of ref document: US