WO2008059368A2 - Fused 2-amino pyrimidine compounds and their use for the treatment of cancer - Google Patents

Fused 2-amino pyrimidine compounds and their use for the treatment of cancer Download PDF

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Publication number
WO2008059368A2
WO2008059368A2 PCT/IB2007/003532 IB2007003532W WO2008059368A2 WO 2008059368 A2 WO2008059368 A2 WO 2008059368A2 IB 2007003532 W IB2007003532 W IB 2007003532W WO 2008059368 A2 WO2008059368 A2 WO 2008059368A2
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Prior art keywords
alkyl
compound
heteroaryl
cycloheteroalkyl
halogen
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PCT/IB2007/003532
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French (fr)
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WO2008059368A3 (en
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Pei-Pei Kung
Jerry Jialun Meng
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Pfizer Products Inc.
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Publication of WO2008059368A3 publication Critical patent/WO2008059368A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • the present invention is directed to compounds, and pharmaceutically acceptable salts and solvates thereof, their synthesis, and their use as modulators or inhibitors of HSP-90.
  • the compounds of the present invention are useful for modulating (e.g. inhibiting) HSP-90 activity and for treating diseases or conditions mediated by HSP-90, such as for example, disease states associated with abnormal cell growth such as cancer.
  • chaperones play important roles in cellular function by ensuring proper folding of proteins upon synthesis as well as their refolding under conditions of denaturing stress. By regulating the balance between protein synthesis and degradation, molecular chaperones are a significant part of the cellular response to stress. In addition, by regulating the proper folding of various cellular proteins, chaperones play an important role in regulating cellular functions such as cell proliferation and apoptosis. (See, e.g. Jolly, et al., J. Natl. Cancer Inst. 92: 1564-1572 (2000)).
  • HSPs Heat shock proteins
  • HSPs are a class of chaperones that accumulate in the cell in response to various environmental stresses, such as heat shock, oxidative stress, or the presence of alcohols or heavy metals.
  • HSPs may also play a significant role as chaperones for a variety of cellular proteins under stress-free conditions.
  • Members of the HSP family are classified according to their molecular weight (e.g. HSP-27, HSP-70, and HSP-90).
  • HSP-27, HSP-70, and HSP-90 are classified according to their molecular weight (e.g. HSP-27, HSP-70, and HSP-90).
  • Evidence of differential expression of HSPs in various stages of tumor progression suggests HSPs play a role in cancer. (See, e.g. Martin, et al., Cancer Res. 60:2232-2238 (2000)).
  • HSP-90 is a homodimer with ATPase activity and functions in a series of complex interactions with a variety of substrate proteins (Young, et al., J. Cell Biol. 154: 267-273 (2001)). HSP-90 is unique with regard to other chaperones, however, since most of its known substrate proteins are signal transduction proteins. Thus, HSP-90 plays an essential role in regulating cellular signal transduction networks. (See, e.g. Xu, et al., Proc. Natl. Acad. Sci 90:7074-7078 (1993)).
  • substrate proteins of HSP-90 include many mutated or over-expressed proteins implicated in cancer such as p53, Bcr-At>1 kinase, Raf-1 kinase, Akt kinase, Npm-Alk kinase p185 ErbB2 transmembrane kinase, Cdk4, Cdk6, Wee1 (a cell cycle-dependent kinase), HER2/Neu (ErbB2), and hypoxia inducible factor-l et (HIF-Ia).
  • HSP-90 results in selective degradation of these important signaling proteins involved in apoptosis, cell proliferation, and cell cycle regulation (Holstein, et al., Cancer Res.
  • HSP-90 is an attractive therapeutic target because of the important roles played by these signaling proteins in disease states involving abnormal cell growth, such as cancer. It is thus desirable to discover and develop new inhibitors of HSP-90 activity that can provide a therapeutic benefit to patients suffering from disease states related to abnormal cell growth such as cancer.
  • Examples of compounds that inhibit HSP-90 include those described in published patent applications WO 2006/079789, WO 2006/008503, WO 2006/090094, and WO 2005/021552.
  • X is CR 2 , N, or NR 2 ;
  • Y is CH or S, provided that when X is CR 2 , Y is S;
  • R 1 is -(CHa) n -(C 6 to C 14 ) aryl, -(CH 2 ) n -(C 2 to C 9 ) heteroaryl, or -(CH 2 ) n -C(O)NR 4a R 4b , wherein each of said (C 6 to Cu) aryl and (C 2 to Cg) heteroaryl is optionally substituted with at least one R 5 group;
  • R 2 is H, -(CHb) n -(Ce to Ci 4 ) aryl, -(CH 2 ) n -(C 2 to C 9 ) heteroaryl, or - (CH 2 ) n -C(O)NR 4a R 4b , wherein each of said (C 6 to C 14 ) aryl and (C 2 to C 9 ) heteroaryl is optionally substituted with at least one R 5 group;
  • R 3 is H, (Ci to C 6 ) alkyl, (C 2 to C 8 ) alkenyl, (C 2 to C 8 ) alkynyl, -(CH 2 ) n -C(O)OR 7 or -(CH 2 ) n -C(O)NR 4a R 4b ;
  • R 4a and R 4b are each independently H, (Ci to C 6 ) alkyl, (C 2 to C 8 ) alkenyl, (C 2 to C 8 ) alkynyl, (C 6 to Cu) aryl, (C 2 to C 9 ) heteroaryl, (C 2 to C 9 ) cycloheteroalkyl, or (C 3 to C 8 ) cycloalkyl, all optionally substituted with at least one R 8 group, or R 4a and R 4b , taken together with the nitrogen atom to which they are bound, form a (C 2 to C 9 ) cycloheteroalkyl group, wherein said (C 2 to C 9 ) cycloheteroalkyl is optionally substituted with at least one R 8 group; each R 5 is independently -OH, (C 1 to C 6 ) alkyl, cyano, halogen, -C(O)NR 4a R 4b , -NR 4a R 4b , (C 1
  • R 7 is H, or (C 1 to C 6 ) alkyl; each R 8 is independently -OH, (C 1 to C 6 ) alkyl, (C 2 to C 8 ) alkenyl, (C 2 to C 8 ) alkynyl, (C 1 to C 8 ) heteroalkyl, or (C 1 to C 8 ) alkoxy; each n is independently 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof.
  • the compound of formula (I) has the following structure:
  • R 1 is (Ce to Ci 4 ) aryl or (C 2 to Cg) heteroaryl, wherein each of said (C ⁇ to Cu) aryl and (C 2 to C 9 ) heteroaryl is optionally substituted with at least one R 5 group.
  • R 1 is C ⁇ aryl, wherein said Ce aryl is optionally substituted with at least one R 5 group.
  • R 3 is H or -CH 3 . In another embodiment, R 3 is -(CH 2 ) n -C(O)NR 4a R 4b . In another embodiment, R 3 is -C(O)NHR 4b .
  • the compound of formula (I) has the following structure:
  • R 5a is (Ci to C 8 ) alkoxy, which is optionally substituted with at least one R 6 group; R 5b is halogen; and R 5G is halogen.
  • the invention relates to the compound or salt shown above, wherein R 5a is -OCH 3 , R 5b is Br, and R 50 is Cl.
  • the compound of formula (I) has the following structure:
  • R is (Ci to C 8 ) alkoxy, which is optionally substituted with at least one R group;
  • R 5b is halogen
  • F 3 r 5c 3 is halogen
  • the invention relates to the compound or salt shown above, wherein R 5a is -OCH 3 , R 5b is Br, and R 5c is Cl.
  • the invention relates to the compound or salt described above, wherein R 1 is (C ⁇ to Ci 4 ) aryl or (C- 2 to C 9 ) heteroaryl, wherein each of said (Ce to Cu) aryl and (C2 to C 9 ) heteroaryl is optionally substituted with at least one R 5 group.
  • R 1 is Ce aryl, wherein said Ce aryl is optionally substituted with at least one R 5 group.
  • R 3 is H or -CH 3 .
  • R 3 is -(CH 2 ) n -C(O)NR 4a R 4b .
  • R 3 is - C(O)NHR 4b .
  • R 5a is (C 1 to C 8 ) alkoxy, which is optionally substituted with at least one R 6 group;
  • R 5b is halogen
  • R 5c is halogen
  • R 5a is -OCH 3
  • R 5b is Br
  • R 5c is Cl
  • the compound or salt described above has the following structure:
  • R 5a is (Ci to Cs) alkoxy, which is optionally substituted with at least one R 6 group;
  • R 5b is halogen; and R 5c is halogen.
  • R 5a is -OCH 3
  • R 5b is Br
  • R 5c is Cl.
  • R 1 is (C 6 to Cu) aryl or (C2 to C 9 ) heteroaryl, wherein each of said (C 6 to Ci 4 ) aryl and (C 2 to C 9 ) heteroaryl is optionally substituted with at least one R 5 group.
  • R 1 is C 6 aryl, wherein said C 6 aryl is optionally substituted with at least one R 5 group.
  • R 3 is H or -CH 3 . In another embodiment, R 3 is -(CH 2 ) n -C(O)NR 4a R 4b . In another embodiment, R 3 is -C(O)NHR 4b .
  • the compound of formula (I) has the following structure:
  • R 5a is (Ci to Cs) alkoxy, which is optionally substituted with at least one R 6 group;
  • R 5b is halogen
  • R 50 is halogen
  • R 5a is -OCH 3
  • R 5b is Br
  • R 5c is Cl
  • the compound of formula (I) has the following structure:
  • R 5a is (Ci to Ce) alkoxy, which is optionally substituted with at least one R 6 group;
  • R 5b is halogen
  • R Sc is halogen
  • R 5a is -OCH 3
  • R 5b is Br
  • R 5 ° is Cl
  • the present invention also provides a compound of formula (II),
  • X is CR 9 or N;
  • R 9 is H, -OH, halogen, (Ci to C 6 ) alkyl, (Ci to C 6 ) alkoxy or (Ci to C 6 ) perfluoroalkyl;
  • R 10 is selected from the group consisting of
  • each R 11a , R 11b and R 11c is independently selected from the group consisting of
  • each R 12 is independently H, (C 1 to C 6 ) alkyl, (C 2 to C 6 ) alkenyl, (C 2 to C 6 ) alkynyl,
  • each R 12 is optionally further substituted by 1-3 R 14 ; each R 13a and R 13b is H, (C 1 to C 6 ) alkyl, (C 2 to C 6 ) alkenyl, (C 2 to C 6 ) alkynyl, (C 1 to C 6 ) perfluoroalkyl, (C 3 to C 8 ) cycloalkyl, (C 2 to C 9 ) cycloheteroalkyl, (C 6 to Cu) aryl, (C 2 to Cg) heteroaryl, -(Ci to C 3 alkylene)-(C 3 to Ce) cyclcoalkyl, -(Ci to C 3 alkylene)- (C 2 to C 9 cycloheteroalkyl), -(Ci to C 3 alkylene)- (C 6 to C 14 aryl) or -(C 1 to C 3 alkylene)- (C 2 to C 9 heteroaryl), wherein each R 12 is optionally further substituted by 1-3 R 14
  • each R 15 is otpioanlly further substituted by 1-3 R 17 ; each R 16a and R 16b is independently H, (Ci to C 6 ) alkyl, (C 2 to C 6 ) alkenyl, (C 2 to C 6 ) alkynyl, (Ci to C 6 ) perfluoroalkyl, (C 3 to Ca) cycloalkyl, (C 2 to Cg) cycloheteroalkyl, (C 6 to Ci 4 ) aryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C 2 to Cg) heteroaryl, (C
  • each R 16a or R 16b is optionally further substituted by 1-3 R 17 ; or, R 16a and R 16b , taken together with the nitrogen atom to which they are bound, form a (C 2 to C 9 ) cycloheteroalkyl group or a (C 2 to Cg) heteroaryl group, wherein said (C 2 to Cg) cycloheteroalkyl and (C 2 to Cg) heteroaryl group is optionally further substituted by 1-3 R 17 ; each R 17 is independenly halogen, -OH 1 cyano, -S(O) 2 -(Ci to C 3 alkyl), (Ci to C 3 ) alkyl, (Ci to C 3 ) perfluoroalkyl (Ci to C 3 ) alkoxy, -C(O)-(Ci-C 6 alkyl), -C(O)-
  • X is CR 9 . More preferably, R 9 is H, -OH or (Ci to C 6 ) alkoxy. Even more preferably, R 9 is -OH or (Ci to C 6 ) alkoxy. E ⁇ ven more preferably, R 9 is (Ci to C 6 ) alkoxy. In another preferred aspect of the compound of formula (II) or salt thereof, X is N.
  • R 10 is
  • each R 13a and R 13b is independently H or (Ci to C 6 ) alky wherein said (Ci to C 6 ) alkyl is optionally further substituted by 1-3 halogen; even more preferably, R 13a is H. Also more preferably, R 13a and R 13b , taken together with the nitrogen atom to which they are bound, form a (C- 2 to C 9 ) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C 2 to Cg) cycloheteroalkyl or (C2 to Cg) heteroaryl group is optionally further substituted by 1-3 R 14 .
  • R 10 is -C(O)-OR 12 . More preferably, R 12 is selected from (Ci to C 6 ) alkyl, the said (Ci to C 6 ) alkyl is optionally further substituted by 1-3 halogen.
  • R 10 is H.
  • R 10 is (Ci to C 6 ) alkyl optionally further substituted by 1-3 halogen.
  • R 11a and R 11 b is independently halogen, Ci to C 6 alkyl or Ci to C ⁇ perfluoroalkyl
  • R 11c is -O- (Ci to C 6 alkylene)-R 15 , -0-(C 2 to C 6 alkenylene)-R 15 or -0-(C 2 to C 6 alkynylene)-R 15
  • R 15 is (C 2 to C 9 ) heteroaryl, (C 3 to Ce) cycloalkyl or (C 2 to C 9 ) heteroaryl
  • R 15 is optionally further substituted by 1-3 R 17 .
  • R 11a and R 11 b is independently halogen
  • R 11c is -O-(Ci to C 6 alkylene)-(C 2 to C 9 heteroaryl), wherein the (C 2 to C 9 ) heteroaryl is optionally further substituted by 1-3 groups selected from halogen and (Ci to C 3 ) alkyl.
  • X is CR 9 or N
  • R 9 is H, -OH, or (C 1 -C 3 alkoxy);
  • R 10 is selected from (a) H, (Ci to C 6 ) alkyl, and (Ci to C 6 ) perfluoroalkyl,
  • R 12 is H or (Ci to C 6 ) alkyl optionally substituted by 1-3 halogen; and (e) -(CH 2 )p-C(O)NR 13a R 13b , wherein each R 13a and R 13b is selected from H, (Ci to C 6 alkyl) optionally substituted by 1-3 halogen, (C 3 to C 8 ) cycloalkyl optionally substituted by 1-3 groups selected from halogen and (Ci to C 3 ) alkyl, or R 13a and R 13b , taken together with the nitrogen atom to which they are bound, form a (C 2 to C 9 ) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C 2 to C9) cycloheteroalkyl or (C 2 to Cg) heteroaryl group is optionally further substituted by 1-3 groups selected from halogen
  • R 11c is -0-(C 1 to C 6 alkylene)-R 15 , -0-(C 2 to C 6 alkenylene)-R 15 or -0-(C 2 to C 6 alkynylene)-R 15 , R 15 is (C 2 to C 9 ) heteroaryl, (C 3 to C 8 ) cycloalkyl or (C 2 to C 9 ) heteroaryl, and R 15 is optionally further substituted by 1-3 R 17 .
  • R 10 is -C(O)NR 13a R 13b , wherein R 13a is H, R 13b is (Ci to C 6 alkyl) optionally substituted by 1-3 halogen, or R 13a and R 13b , taken together with the nitrogen atom to which they are bound, form a (C 2 to C 9 ) cycloheteroalkyl group, the said (C 2 to Cg) cycloheteroalkyl group is optionally further substituted by 1-3 groups selected from halogen, (Ci to C 3 ) alkyl, (CrC 3 ) perfluoroalkyl and -S(O) 2 -(Ci to C 3 alkyl); each R 11a and R 11b is independently halogen, (Ci to C 6 alkyl) or (Ci to C 6 ) alkoxy;
  • R 110 is -O-(Ci to C 6 alkylene)-R 15 or -0-(C 2 to C 6 alkenylene)-R 15 , R 15 is (C 2 to C 9 ) heteroaryl, and R 15 is optionally further substituted by 1-3 groups seleted from halogen and (Ci to C 3 ) alkyl.
  • the present invention encompasses compounds of formula (II) as described above formed by any and all combinations of the preferred aspects, more preferred aspects, and even more preferred aspects of the compounds of formula (II), wherein any of the value of R 9 , R 10 , R 11a , R 11b , R 11 °, R12, R 13a , R 13b , R 14 , R 15 , R 16a , R 16b and R 17 are further narrowed, so long as the aspects are not inconsistent with each other, and includes pharmaceutically acceptable salts and solvates thereof of any of said compounds.
  • a pharmaceutical composition comprising a compound of formula (II) or a salt thereof.
  • the present invention further provides a pharmaceutical composition, comprising at least one compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier or diluent.
  • the present invention further provides a method of reducing abnormal cell growth in a mammal in need thereof, comprising the step of administering to said mammal a therapeutically effective amount of at least one compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the abnormal cell growth is cancerous.
  • the present invention further provides a method of treating cancer in a mammal, comprising the step of administering to said mammal a therapeutically effective amount of at least one compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the present invention further provides a method of inhibiting HSP-90 enzymatic activity, comprising contacting a HSP-90 enzyme with an HSP-90-inhibiting amount of at least one compound according to any one of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof.
  • the present invention further provides the use of a compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for the treatment of abnormal cell growth in a mammal. In one embodiment, such abnormal cell growth is cancerous.
  • halo and/or halogen refer to fluorine, chlorine, bromine or iodine.
  • (Ci to C 6 ) alkyl refers to a saturated aliphatic hydrocarbon radical including straight chain and branched chain groups of 1 to 6 carbon atoms. Examples of (Ci to C 6 ) alkyl groups include methyl, ethyl, propyl, 2-propyl, n-butyl, /so-butyl, tert- butyl, pentyl, and the like.
  • perfluoroalkyl refers to an alkyl group in which all of its hydrogen atoms are replaced by fluorine atoms.
  • (C 2 to Ce) alkenyl as used herein, means an alkyl moiety comprising
  • (C2 to Cs) alkynyl means an alkyl moiety comprising from 2 to 8 carbon atoms and having at least one carbon-carbon triple bond.
  • the carbon-carbon triple bond in such a group may be anywhere along the 2 to 8 carbon chain that will result in a stable compound. Examples of such groups include, but are not limited to, ethyne, propyne, 1-butyne, 2-butyne, 1-pentyne, 2-pentyne, 1-hexyne, 2- hexyne, and 3-hexyne.
  • (Ci to Cs) alkoxy means an O-alkyl group wherein said alkyl group contains from 1 to 8 carbon atoms and is straight, branched, or cyclic. Examples of such groups include, but are not limited to, methoxy, ethoxy, n-propyloxy, iso-propyloxy, n-butoxy, iso-butoxy, tert-butoxy, cyclopentyloxy, and cyclohexyloxy.
  • (Ci to C 8 ) heteroalkyl refers to a straight- or branched-chain alkyl group having a total of from 2 to 12 atoms in the chain, including from 1 to 8 carbon atoms, and one or more atoms of which is a heteroatom selected from S, O, and N, with the proviso that said chain may not contain two adjacent O atoms or two adjacent S atoms.
  • the S atoms in said chains may be optionally oxidized with one or two oxygen atoms, to afford sulfides and sulfones, respectively.
  • the (Ci to Ce) heteroalkyl groups in the compounds of the present invention can contain an oxo group at any carbon or heteroatom that will result in a stable compound.
  • Exemplary (Ci to C 3 ) heteroalkyl groups include, but are not limited to, alcohols, alkyl ethers, primary, secondary, and tertiary alkyl amines, amides, ketones, esters, sulfides, and sulfones.
  • ⁇ CQ to Cu) aryl means a group derived from an aromatic hydrocarbon containing from 6 to 14 carbon atoms. Examples of such groups include, but are not limited to, phenyl or naphthyl.
  • Ph and phenyl
  • benzyl as used herein, means a -CH 2 C 6 H 5 group.
  • (C 2 to Cg) heteroaryl means an aromatic heterocyclic group having a total of from 5 to 10 atoms in its ring, and containing from 2 to 9 carbon atoms and from one to four heteroatoms each independently selected from O, S and N, and with the proviso that the ring of said group does not contain two adjacent O atoms or two adjacent S atoms.
  • the heterocyclic groups include benzo-fused ring systems.
  • aromatic heterocyclic groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furaza ⁇ yl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinox
  • the C 2 to Cg heteroaryl groups may be C-attached or N-attached where such is possible.
  • a group derived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached).
  • a group derived from imidazole may be imidazol-1-yl (N-attached) or imidazol-3-yl (O attached).
  • (C-2 to Cg) cycloheteroalkyl means a non-aromatic, monocyclic, bicyclic, tricyclic, spirocyclic, or tetracyclic group having a total of from 4 to 13 atoms in its ring system, and containing from 2 to 9 carbon atoms and from 1 to 4 heteroatoms each independently selected from O, S and N, and with the proviso that the ring of said group does not contain two adjacent O atoms or two adjacent S atoms.
  • such C 2 to Cg cycloheteroalkyl groups may contain an oxo substituent at any available atom that will result in a stable compound.
  • such a group may contain an oxo atom at an available carbon or nitrogen atom. Such a group may contain more than one oxo substituent if chemically feasible.
  • a C 2 to C 9 cycloheteroalkyl group contains a sulfur atom, said sulfur atom may be oxidized with one or two oxygen atoms to afford either a sulfoxide or sulfone.
  • An example of a 4 membered cycloheteroalkyl group is azetidinyl (derived from azetidine).
  • An example of a 5 membered cycloheteroalkyl group is pyrrolidinyl.
  • An example of a 6 membered cycloheteroalkyl group is piperidinyl.
  • An example of a 9 membered cycloheteroalkyl group is indolinyl.
  • An example of a 10 membered cycloheteroalkyl group is 4H- quinolizinyl.
  • C 2 to C 9 cycloheteroalkyl groups include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1 ,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1 ,3-dioxolanyl
  • (C 3 to C 10 ) cycloalkyl group means a saturated or unsaturated, monocyclic, fused, spirocyclic, or polycyclic ring structure having a total of from 3 to 8 carbon ring atoms.
  • groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, and adamantyl.
  • (C 3 to C 6 ) cycloalkyl or “(C 4 to C 8 ) cycloalkyl” refer to the corresponding saturated or unsaturated, monocyclic, fused, spirocyclic, or polycyclic ring structure having a total of from 3 to 6 carbon ring atoms or from 4 to 8 carbon atoms, respectively.
  • cyano refers to a -C ⁇ N group.
  • the new term refers to a diradical formed by removing one hydrogen atom from the original term of which the new term derived from.
  • an alkylene refers to a diradical group formed by removing one hydrogen atom from an alkyl group and that a "methylene” refers to a divalent radical -CH 2 - derived from removing one hydrogen atom from methyl.
  • diradicals include, but are not limited to: alkenylene, alkynylene, cycloalkylene, phenylene, heterocyclylene, heteroarylene and (nonaromatic unsaturated carbocyclylene), which are derived from alkenyl, alkynyl, cycloalkyl, phenyl, heterocyclyl, heteroary! and (nonaromatic unsaturated carbocyclyl), respectively.
  • cyclopropylene For example, “CrC 2 alkylene” refers to all of the following: -CH 2 -, -CH(CH 3 )- and -CH 2 -CH 2 -.
  • substituted means that the specified group or moiety bears one or more substituents.
  • unsubstituted means that the specified group bears no substituents.
  • optionally substituted means that the specified group is unsubstituted or substituted by one or more substituents. It is to be understood that in the compounds of the present invention when a group is said to be “unsubstituted,” or is “substituted” with fewer groups than would fill the valencies of all the atoms in the compound, the remaining valencies on such a group are filled by hydrogen.
  • Ce aryl group also called "phenyl” herein
  • phenyl substituted with one additional substituent
  • one of ordinary skill in the art would understand that such a group has 4 open positions left on carbon atoms of the Ce aryl ring (6 initial positions, minus one to which the remainder of the compound of the present invention is bonded, minus an additional substituent, to leave 4).
  • the remaining 4 carbon atoms are each bound to one hydrogen atom to fill their valencies.
  • a C ⁇ aryl group in the present compounds is said to be "disubstituted," one of ordinary skill in the art would understand it to mean that the Ce aryl has 3 carbon atoms remaining that are unsubstituted.
  • solvate means a pharmaceutically acceptable solvate form of a compound of the present invention that retains the biological effectiveness of such compound.
  • solvates include, but are not limited to, compounds of the invention in combination with water, isopropanol, ethanol, methanol, dimethylsulfoxide (DMSO), ethyl acetate, acetic acid, ethanolamine, or mixtures thereof. It is specifically contemplated that in the present invention one solvent molecule can be associated with one molecule of the compounds of the present invention, such as a hydrate.
  • solvates of the present invention are contemplated as solvates of compounds of the present invention that retain the biological effectiveness of the non-hydrate form of the compounds.
  • pharmaceutically acceptable salt means a salt of a compound of the present invention that retains the biological effectiveness of the free acids and bases of the specified derivative and that is not biologically or otherwise undesirable.
  • pharmaceutically acceptable formulation means a combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a carrier, diluent, and/or excipient(s) that are compatible with a compound of the present invention, and is not deleterious to the recipient thereof.
  • Pharmaceutical formulations can be prepared by procedures known to those of ordinary skill in the art.
  • the compounds of the present invention can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, and the like.
  • excipients, diluents, and carriers that are suitable for such formulations include the following: fillers and extenders such as starch, sugars, mannitol, and silicic derivatives; binding agents such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl pyrrolidone; moisturizing agents such as glycerol; disintegrating agents such as povidone, sodium starch glycolate, sodium carboxymethylcellulose, agar, calcium carbonate, and sodium bicarbonate; agents for retarding dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as cetyl alcohol, glycerol monostearate; adsorptive carriers such as kaolin and bentonite; and lubricants such as talc, calcium and magnesium stearate and solid polyethylene glycols.
  • fillers and extenders such as starch, sugars, mannitol, and silicic derivatives
  • binding agents such
  • Final pharmaceutical forms may be pills, tablets, powders, lozenges, saches, cachets, or sterile packaged powders, and the like, depending on the type of excipient used. Additionally, it is specifically contemplated that pharmaceutically acceptable formulations of the present invention can contain more than one active ingredient. For example, such formulations may contain more than one compound according to the present invention. Alternatively, such formulations may contain one or more compounds of the present invention and one or more additional agents that reduce abnormal cell growth.
  • HSP-90-inhibiting amount refers to the amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, required to inhibit the enzymatic activity of HSP-90 in vivo, such as in a mammal, or in vitro.
  • the amount of such compounds required to cause such inhibition can be determined without undue experimentation using methods described herein and those known to those of ordinary skill in the art.
  • inhibiting HSP-90 enzyme activity means decreasing the activity or functioning of the HSP-90 enzyme either in vitro or in vivo, such as in a mammal, such as a human, by contacting the enzyme with a compound of the present invention.
  • a therapeutically effective amount means an amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, that, when administered to a mammal in need of such treatment, is sufficient to effect treatment, as defined herein.
  • a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof is a quantity sufficient to modulate or inhibit the activity of the HSP-90 enzyme such that a disease condition that is mediated by activity of the HSP-90 enzyme is reduced or alleviated.
  • treat refers to any treatment of an HSP-90 mediated disease or condition in a mammal, particularly a human, and include: (i) preventing the disease or condition from occurring in a subject which may be predisposed to the condition, such that the treatment constitutes prophylactic treatment for the pathologic condition; (ii) modulating or inhibiting the disease or condition, i.e., arresting its development; (iii) relieving the disease or condition, i.e., causing regression of the disease or condition; or (iv) relieving and/or alleviating the disease or condition or the symptoms resulting from the disease or condition, e.g., relieving an inflammatory response without addressing the underlying disease or condition.
  • these terms simply mean that the life expectancy of an individual affected with a cancer will be increased or that one or more of the symptoms of the disease will be reduced.
  • compound of the present invention refers to any of the above- mentioned compounds, as well as those in the Examples that follow, and include those generically described or those described as species.
  • the term also refers to pharmaceutically acceptable salts or solvates of these compounds.
  • abnormal cell growth refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition), including the abnormal growth of normal cells and the growth of abnormal cells. This includes, but is not limited to, the abnormal growth of: tumor cells (tumors) that proliferate by expressing a mutated tyrosine kinase or overexpression of a receptor tyrosine kinase; benign and malignant cells of other proliferative diseases in which aberrant tyrosine kinase activation occurs; any tumors that proliferate by receptor tyrosine kinases; any tumors that proliferate by aberrant serine/threonine kinase activation; benign and malignant cells of other proliferative diseases in which aberrant serine/threonine kinase activation occurs; tumors, both benign and malignant, expressing an activated Ras oncogene; tumor cells, both benign and malignant, in which the Ras protein
  • abnormal cell growth also refers to and includes the abnormal growth of cells, both benign and malignant, resulting from activity of the enzyme famesyl protein transferase.
  • abnormal cell growth and “hyperproliferative disorder” are used interchangeably in this application.
  • stereoisomers refers to compounds that have identical chemical constitution, but differ with regard to the arrangement of their atoms or groups in space.
  • enantiomers refers to two stereoisomers of a compound that are non-superimposable mirror images of one another.
  • racemic or “racemic mixture,” as used herein, refer to a 1 :1 mixture of enantiomers of a particular compound.
  • diastereomers refers to the relationship between a pair of stereoisomers that comprise two or more asymmetric centers and are not mirror images of one another.
  • HSP-90 activity is useful for the prevention and/or treatment of disease states associated with abnormal cell growth such as cancer, alone or in combination with other anti-cancer agents.
  • hydrogen atoms are not explicitly depicted, e.g., ' ⁇ - represents a methyl group,
  • the compounds of the present invention may have asymmetric carbon atoms.
  • the carbon-carbon bonds of the compounds of the present invention may be depicted herein using a solid line ( ), a solid wedge ( m ), or a dotted wedge (--"" 111 Hi).
  • a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that all possible stereoisomers (e.g. specific enantiomers, racemic mixtures, etc.) at that carbon atom are included.
  • the use of either a solid or dotted wedge to depict bonds to asymmetric carbon atoms is meant to indicate that only the stereoisomer shown is meant to be included. It is possible that compounds of the invention may contain more than one asymmetric carbon atom. In those compounds, the use of a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that all possible stereoisomers are meant to be included.
  • the compounds of the present invention can exist as enantiomers and diastereomers or as racemates and mixtures thereof.
  • the use of a solid line to depict bonds to one or more asymmetric carbon atoms in a compound of the invention and the use of a solid or dotted wedge to depict bonds to other asymmetric carbon atoms in the same compound is meant to indicate that a mixture of diastereomers is present.
  • enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate using, for example, chiral high pressure liquid chromatography (HPLC).
  • HPLC high pressure liquid chromatography
  • the racemate (or a racemic precursor) may be reacted with a suitable optically active compound, for example, an alcohol, or, in the case where the compound contains an acidic or basic moiety, an acid or base such as tartaric acid or 1- phenylethylamine.
  • a suitable optically active compound for example, an alcohol, or, in the case where the compound contains an acidic or basic moiety, an acid or base such as tartaric acid or 1- phenylethylamine.
  • the resulting diastereomeric mixture may be separated by chromatography and/or fractional crystallization and one or both of the diastereoisomers converted to the corresponding pure enantiomer(s) by means well known to one skilled in the art.
  • Chiral compounds of the invention may be obtained in enantiomerically-enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing from 0 to 50% isopropanol, typically from 2 to 20%, and from 0 to 5% of an alkylamine, typically 0.1% diethylamine. Concentration of the eluate affords the enriched mixture.
  • Stereoisomeric conglomerates may be separated by conventional techniques known to those skilled in the art. See, e.g. "Stereochemistry of Organic Compounds" by E L Eliel (Wiley, New York, 1994), the disclosure of which is incorporated herein by reference in its entirety.
  • a compound of the invention contains an alkenyl or alkenylene group
  • geometric cisltrans (or ZIE) isomers are possible.
  • the compound contains, for example, a keto or oxime group or an aromatic moiety
  • tautomeric isomerism ('tautomerism') can occur.
  • Examples of tautomerism include keto and enol tautomers.
  • a single compound may exhibit more than one type of isomerism. Included within the scope of the invention are all stereoisomers, geometric isomers and tautomeric forms of the inventive compounds, including compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof.
  • Cis/trans isomers may be separated by conventional techniques well known to those skilled in the art, for example, chromatography and fractional crystallization.
  • Salts of the present invention can be prepared according to methods known to those of skill in the art.
  • Examples of salts include, but are not limited to, acetate, acrylate, benzenesulfonate, benzoate (such as chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, and methoxybenzoate), bicarbonate, bisulfate, bisulfite, bitartrate, borate, bromide, butyne-1 ,4-dioate, calcium edetate, camsylate, carbonate, chloride, caproate, caprylate, clavula ⁇ ate, citrate, decanoate, dihydrochloride, dihydrogenphosphate, edetate, edislyate, estolate, esylate, ethylsuccinate, formate, fumarate, gluceptate, gluconate, glutamate, glycollate, glycollylarsanilate, heptanoate,
  • the compounds of the present invention that are basic in nature are capable of forming a wide variety of different salts with various inorganic and organic acids. Although such salts must be pharmaceutically acceptable for administration to animals, it is often desirable in practice to initially isolate the compound of the present invention from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert the latter back to the free base compound by treatment with an alkaline reagent and subsequently convert the latter free base to a pharmaceutically acceptable acid addition salt.
  • the acid addition salts of the base compounds of this invention can be prepared by treating the base compound with a substantially equivalent amount of the selected mineral or organic acid in an aqueous solvent medium or in a suitable organic solvent, such as methanol or ethanol. Upon evaporation of the solvent, the desired solid salt is obtained.
  • the desired acid salt can also be precipitated from a solution of the free base in an organic solvent by adding an appropriate mineral or organic acid to the solution.
  • Those compounds of the present invention that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations.
  • Examples of such salts include the alkali metal or alkaline-earth metal salts and particularly, the sodium and potassium salts. These salts are all prepared by conventional techniques.
  • the chemical bases which are used as reagents to prepare the pharmaceutically acceptable base salts of this invention are those which form non-toxic base salts with the acidic compounds of the present invention.
  • Such non-toxic base salts include those derived from such pharmacologically acceptable cations as sodium, potassium calcium and magnesium, etc.
  • salts can be prepared by treating the corresponding acidic compounds with an aqueous solution containing the desired pharmacologically acceptable cations, and then evaporating the resulting solution to dryness, preferably under reduced pressure.
  • they may also be prepared by mixing lower alkanolic solutions of the acidic compounds and the desired alkali metal alkoxide together, and then evaporating the resulting solution to dryness in the same manner as before.
  • stoichiometric quantities of reagents are preferably employed in order to ensure completeness of reaction and maximum yields of the desired final product.
  • the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.
  • an inorganic acid such as hydrochloric acid
  • the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like.
  • suitable salts include organic salts derived from amino acids, such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.
  • inventive compounds, agents and salts may exist in different crystal or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulas.
  • the invention also includes isotopically-labeled compounds of the invention, wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as 2 H and 3 H, carbon, such as 11 C, 13 C and 14 C, chlorine, such as 36 CI, fluorine, such as 18 F, iodine, such as 123 I and 125 I, nitrogen, such as 13 N and 15 N, oxygen, such as 15 O, 17 O and 18 O, phosphorus, such as 32 P, and sulfur, such as 35 S.
  • isotopically-labeled compounds of the invention for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies.
  • the radioactive isotopes tritium, 3 H, and carbon-14, U C are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium, 2 H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as 11 C, 18 F, 15 O and 13 N 1 can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy.
  • PET Positron Emission Topography
  • Isotopically-labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed.
  • compositions of the invention comprise a therapeutically effective amount of at least one compound of the present invention and an inert, pharmaceutically acceptable carrier or diluent.
  • a pharmaceutical composition of the invention is administered in a suitable formulation prepared by combining a therapeutically effective amount (i.e., an HSP-90 modulating, regulating, or inhibiting amount effective to achieve therapeutic efficacy) of at least one compound of the present invention (as an active ingredient) with one or more pharmaceutically suitable carriers, which may be selected, for example, from diluents, excipients and auxiliaries that facilitate processing of the active compounds into the final pharmaceutical preparations.
  • a therapeutically effective amount i.e., an HSP-90 modulating, regulating, or inhibiting amount effective to achieve therapeutic efficacy
  • one compound of the present invention as an active ingredient
  • pharmaceutically suitable carriers which may be selected, for example, from diluents, excipients and auxiliaries that facilitate processing of the active compounds into the final pharmaceutical preparations.
  • the pharmaceutical carriers employed may be either solid or liquid.
  • Exemplary solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like.
  • Exemplary liquid carriers are syrup, peanut oil, olive oil, water and the like.
  • the inventive compositions may include time-delay or time-release material known in the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate or the like. Further additives or excipients may be added to achieve the desired formulation properties.
  • a bioavailability enhancer such as Labrasol, Gelucire or the like, or formulator, such as CMC (carboxy-methylcellulose), PG (propyleneglycol), or PEG (polyethyleneglycol), may be added.
  • CMC carboxy-methylcellulose
  • PG propyleneglycol
  • PEG polyethyleneglycol
  • Gelucire ® a semisolid vehicle that protects active ingredients from light, moisture and oxidation, may be added, e.g., when preparing a capsule formulation.
  • the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form, or formed into a troche or lozenge.
  • the amount of solid carrier may vary, but generally will be from about 25 mg to about 1 g.
  • the preparation may be in the form of syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampoule or vial or non-aqueous liquid suspension.
  • a semi-solid carrier is used, the preparation may be in the form of hard and soft gelatin capsule formulations.
  • the inventive compositions are prepared in unit-dosage form appropriate for the mode of administration, e.g., parenteral or oral administration.
  • a pharmaceutically acceptable salt of a compound of the present invention may be dissolved in an aqueous solution of an organic or inorganic acid, such as a 0.3 M solution of succinic acid or citric acid.
  • the agent may be dissolved in a suitable co-solvent or combinations of co-solvents.
  • suitable co-solvents include alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerin and the like in concentrations ranging from 0-60% of the total volume.
  • a compound of Formula I is dissolved in DMSO and diluted with water.
  • the composition may also be in the form of a solution of a salt form of the active ingredient in an appropriate aqueous vehicle such as water or isotonic saline or dextrose solution.
  • the agents of the compounds of the present invention may be formulated into aqueous solutions, preferably in physiologically compatible buffers such as Hanks solution, Ringer's solution, or physiological saline buffer.
  • physiologically compatible buffers such as Hanks solution, Ringer's solution, or physiological saline buffer.
  • penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
  • the compounds can be formulated by combining the active compounds with pharmaceutically acceptable carriers known in the art.
  • Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated.
  • Pharmaceutical preparations for oral use can be obtained using a solid excipient in admixture with the active ingredient (agent), optionally grinding the resulting mixture, and processing the mixture of granules after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
  • Suitable excipients include: fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; and cellulose preparations, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP).
  • PVP polyvinylpyrrolidone
  • disintegrating agents may be added, such as crosslinked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
  • Dragee cores are provided with suitable coatings.
  • suitable coatings For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, polyvinyl pyrrolidone, Carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
  • Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active agents.
  • Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules can contain the active ingredients in admixture with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate, and, optionally, stabilizers.
  • the active agents may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • suitable liquids such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration.
  • the compositions may take the form of tablets or lozenges formulated in conventional manner.
  • the compounds for use according to the present invention may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • a suitable propellant e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • Capsules and cartridges of gelatin for use in an inhaler or insufflator and the like may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
  • the compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion.
  • Formulations for injection may be presented in unit-dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative.
  • the compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • compositions for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active agents may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
  • the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
  • a suitable vehicle e.g., sterile pyrogen-free water
  • the compounds of the present invention may also be formulated as a depot preparation. Such long-acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection.
  • the compounds may be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion-exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
  • a pharmaceutical carrier for hydrophobic compounds is a co- solvent system comprising benzyl alcohol, a non-polar surfactant, a water-miscible organic polymer, and an aqueous phase.
  • the co-solvent system may be a VPD co- solvent system.
  • VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the non-polar surfactant polysorbate 80, and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol.
  • the VPD co-solvent system (VPD: 5W) contains VPD diluted 1 :1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration.
  • the proportions of a co-solvent system may be suitably varied without destroying its solubility and toxicity characteristics.
  • identity of the co-solvent components may be varied: for example, other low-toxicity non-polar surfactants may be used instead of polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars or polysaccharides may be substituted for dextrose.
  • other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs.
  • Certain organic solvents such as dimethylsulfoxide also may be employed, although usually at the cost of greater toxicity due to the toxic nature of DMSO.
  • the compounds may be delivered using a sustained- release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent.
  • sustained-release materials have been established and are known by those skilled in the art.
  • Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days.
  • additional strategies for protein stabilization may be employed.
  • the pharmaceutical compositions also may comprise suitable solid- or gel-phase carriers or excipients.
  • These carriers and excipients may provide marked improvement in the bioavailability of poorly soluble drugs.
  • Examples of such carriers or excipients include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.
  • additives or excipients such as Gelucire®, Capryol®, Labrafil®, Labrasol®, Lauroglycol®, Plurol®, Peceol® Transcutol® and the like may be used.
  • the pharmaceutical composition may be incorporated into a skin patch for delivery of the drug directly onto the skin.
  • an exemplary daily dose generally employed will be from about 0.001 to about 1000 mg/kg of body weight, with courses of treatment repeated at appropriate intervals.
  • the pharmaceutically acceptable formulations of the present invention may contain a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, in an amount of about 0.1 mg to about 2000 mg, or from about 1 mg to about 1500 mg, or from about 5 mg to about 1000 mg, or from about 10 mg to about 750 mg, or from about 20 mg to about 500 mg, or from about 30 mg to about 500 mg, or from about 50 to about 500 mg, or from about 100 mg to about 500mg.
  • the pharmaceutically acceptable formulations of the present invention may contain a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, in an amount from about 0.5 w/w% to about 95 w/w%, or from about 1 w/w% to about 95 w/w%, or from about 1 w/w% to about 75 w/w%, or from about 5 w/w% to about 75 w/w%, or from about 10 w/w% to about 75 w/w%, or from about 10 w/w% to about 50 w/w%.
  • the compounds of the present invention may be administered to a mammal suffering from abnormal cell growth, such as a human, either alone or as part of a pharmaceutically acceptable formulation, once a day, twice a day, three times a day, or four times a day, or even more frequently.
  • This invention also relates to a method for the treatment of abnormal cell growth in a mammal, including a human, comprising administering to said mammal an amount of any of the compound described herein, or a pharmaceutically acceptable salt or solvate thereof, that is effective in treating abnormal cell growth.
  • the abnormal cell growth is cancer, including, but not limited to, mesothelioma, hepatobilliary (hepatic and billiary duct), a primary or secondary CNS tumor, a primary or secondary brain tumor, lung cancer (NSCLC and SCLC), bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, gastrointestinal (gastric, colorectal, and duodenal), breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of
  • the cancer is selected from lung cancer (NSCLC and SCLC), cancer of the head or neck, ovarian cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, breast cancer, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), primary CNS lymphoma, non hodgkins's lymphoma, spinal axis tumors, or a combination of one or more of the foregoing cancers.
  • lung cancer NSCLC and SCLC
  • SCLC central nervous system
  • said abnormal cell growth is a benign proliferative disease, including, but not limited to, psoriasis, benign prostatic hypertrophy or restinosis.
  • This invention also relates to a method for the treatment of abnormal cell growth in a mammal which comprises administering to said mammal an amount of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, that is effective in treating abnormal cell growth in combination with an anti-tumor agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxics, anti-hormones, and anti-androgens.
  • an anti-tumor agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxics, anti-hormones, and anti-androgens.
  • the anti-tumor agent used in conjunction with a compound of the present invention and pharmaceutical compositions described herein is an anti-angiogenesis agent, kinase inhibitor, pan kinase inhibitor or growth factor inhibitor.
  • Preferred pan kinase inhibitors include SutentTM (sunitinib), described in U.S. Patent No. 6,573,293.
  • Anti-angiogenesis agents include but are not limited to the following agents, such as EGF inhibitors, EGFR inhibitors, VEGF inhibitors, VEGFR inhibitors, TIE2 inhibitors, IGF1R inhibitors, COX-II (cyclooxygenase II) inhibitors, MMP-2 (matrix-metalloprotienase 2) inhibitors, and MMP-9 (matrix- metalloprotienase 9) inhibitors.
  • VEGF inhibitors include for example, Avastin (bevacizumab), an anti- VEGF monoclonal antibody of Genentech, Inc. of South San Francisco, California. Additional VEGF inhibitors include CP-547,632 (Pfizer Inc., NY, USA), AG13736 (Pfizer Inc.), ZD-6474 (AstraZeneca), AEE788 (Novartis), AZD-2171 , VEGF Trap (Regeneron/Aventis), Vatalanib (also known as PTK-787, ZK-222584: Novartis & Schering AG), Macugen (pegaptanib octasodium, NX-1838, EYE-001 , Pfizer In ⁇ /Gilead/Eyetech), IM862 (Cytran Inc. of Kirkland, Washington, USA); and a ⁇ giozyme, a synthetic ribozyme from Ribozyme (Boulder, Colorado) and Chiron (
  • VEGF inhibitors useful in the practice of the present invention are described in U.S. Patent No. 6,534,524 and 6,235,764, both of which are incorporated in their entirety for all purposes. Additional VEGF inhibitors are described in, for example in WO 99/24440, in WO 95/21613, WO 99/61422, U.S. Patent 5,834,504, WO 98/50356, U.S. Patent 5,883,113 U.S. Patent 5,886,020, U.S. Patent 5,792,783, U.S.
  • anti-angiogenic compounds include acitretin, fenretinide, thalidomide, zoledronic acid, angiostatin, aplidine, cilengtide, combretastatin A-4, endostatin, halofuginone, rebimastat, removab, Revlimid, squalamine, ukrain, Vitaxin and combinations thereof.
  • antiproliferative agents that may be used in combination with the compounds of the present invention include inhibitors of the enzyme farnesyl protein transferase and inhibitors of the receptor tyrosine kinase PDGFr, including the compounds disclosed and claimed in the following: U.S. Patent 6,080,769; U.S. Patent
  • PDGRr inhibitors include but are not limited to those disclosed in international patent application publication numbers WO 01/40217 and WO 04/020431 , the contents of which are incorporated in their entirety for all purposes.
  • Preferred PDGFr inhibitors include Pfizer's CP-673,451 and CP-868,596 and its salts.
  • Preferred GARF inhibitors include Pfizer's AG-2037 (pelitrexol and its salts).
  • GARF inhibitors useful in the practice of the present invention are disclosed in U.S. Patent 5,608,082 which is incorporated in its entirety for all purposes.
  • Examples of useful COX-II inhibitors which can be used in conjunction with a compound of Formula (I) and pharmaceutical compositions disclosed herein include CELEBREXTM (celecoxib), parecoxib, deracoxib, ABT-963, MK-663 (etoricoxib), COX- 189 (Lumiracoxib), BMS 347070, RS 57067, NS-398, Bextra (valdecoxib), paracoxib, Vioxx (rofecoxib), SD-8381, 4-Methyl-2-(3,4-dimethylphenyl)-1-(4-sulfamoyl-phenyl)-1H- pyrrole, 2-(4-Ethoxyphenyl)-4-methyl-1-(4-sulfamoylphenyl)-1H-pyrrole, T-614, J
  • COX-II inhibitors are disclosed in U.S. patent applications US 2005-0148627 and US 2005- 0148777, the contents of which are incorporated in their entirety for all purposes.
  • the anti-tumor agent is celecoxib (U.S. Patent
  • Patent 6,180,651 NS-398 (CAS 123653-11-2), RS 57067 (CAS 17932- 91-3), 4-Methyl-2-(3,4-dimethylphenyl)-1-(4-sulfamoyl-phenyl)-1 H-pyrrole, 2-(4- Ethoxyphenyl)-4-methyl-1-(4-sulfamoylphenyl)-1 H-pyrrole, or meloxicam.
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs
  • NSAIDs non-steroidal anti-inflammatory drugs which inhibit the enzyme that makes prostaglandins (cyclooxygenase I and II), resulting in lower levels of prostaglandins
  • Salsalate Amigesic
  • Diflunisal Dolobid
  • lbuprofen Metoprofen
  • Orudis Nabumetone
  • Relafen Piroxicam
  • Naproxen Aleve, Naprosyn
  • Diclofenac Voltaren
  • lndomethacin Indocin
  • Sulindac Cl
  • Preferred COX-I inhibitors include ib ⁇ profen (Motrin), n ⁇ prin, naproxen (Aleve), indomethacin (Indocin), nabumetone (Relafen) and combinations thereof.
  • EGFr inhibitors such as lressa (gefitinib, AstraZeneca), Tarceva (erlotinib or OSI-774, OSI Pharmaceuticals Inc.), Erbitux (cetuximab, lmclone Pharmaceuticals, Inc.), EMD-7200 (Merck AG), ABX-EGF (Amgen Inc. and Abgenix Inc.), HR3 (Cuban Government), IgA antibodies (University of Er Weg-Nuremberg), TP-38 (IVAX), EGFR fusion protein, EGF-vaccine, anti-EGFr immunoliposomes (Hermes Biosciences Inc.) and combinations thereof.
  • Preferred EGFr inhibitors include Iressa, Erbitux, Tarceva and combinations thereof.
  • Other anti-tumor agents include those selected from pan erb receptor inhibitors or
  • ErbB2 receptor inhibitors such as CP-724,714 (Pfizer, Inc.), CM 033 (canertinib, Pfizer, Inc.), Herceptin (trastuzumab, Genentech Inc.), Omitarg (2C4, pertuzumab, Genentech Inc.), TAK-165 (Takeda), GW-572016 (lonafamib, GlaxoSmithKline), GW-282974 (GlaxoSmithKline), EKB-569 (Wyeth), PKI-166 (Novartis), dHER2 (HER2 Vaccine, Corixa and GlaxoSmithKline), APC8024 (HER2 Vaccine, Dendreon), anti-HER2/neu bispecific antibody (Decof Cancer Center), B7.her2.lgG3 (Agensys), AS HER2 (Research Institute for Rad Biology & Medicine), trifunctional bispecific antibodies (University of Munich) and mAB AR-209 (Aronex Pharmaceuticals Inc)
  • pan erbb receptor inhibitors include GW572016, CI-1033, EKB-569, and Omitarg and combinations thereof.
  • Additional erbB2 inhibitors include those disclosed in WO 98/02434, WO 99/35146, WO 99/35132, WO 98/02437, WO 97/13760, WO 95/19970, U.S. Patent 5,587,458, and U.S. Patent 5,877,305, each of which is herein incorporated by reference in its entirety.
  • ErbB2 receptor inhibitors useful in the present invention are also disclosed in U.S. Patents 6,465,449, and 6,284,764, and in WO 01/98277 each of which are herein incorporated by reference in their entirety.
  • other anti-tumor agents may be selected from the following agents,
  • BAY-43-9006 Onyx Pharmaceuticals Inc.), Genasense (augmerosen, Genta), Panitumumab (Abgenix/Amgen), Zevalin (Schering), Bexxar (Corixa/GlaxoSmithKline), Abarelix, Alimta, EPO 906 (Novartis), discodermolide (XAA-296), ABT-510 (Abbott), Neovastat (Aeterna), enzastaurin (EIi Lilly), Combrestatin A4P (Oxigene), 2D-6126 (AstraZeneca), flavopiridol (Aventis), CYC-202 (Cyclacel), AVE-8062 (Aventis), DMXAA (Roche/Antisoma), Thymitaq (Eximias), Temodar (temozolomide, Schering Plough) and Revilimd (Celegene) and combinations thereof.
  • Other anti-tumor agents may be selected from the following agents, Cy
  • ABT-627 Satraplatin (JM-216), thalomid (Thalidomide), Theratope, Temilifene (DPPE),
  • ABI-007 paclitaxel
  • Evista raloxifene
  • Atamestane Biomed-777
  • Xyotax polyglutamate paclitaxel
  • Targetin bexarotine
  • anti-tumor agents may be selected from the following agents, Trizaone (tirapazamine), Aposyn (exisulind), Nevastat (AE-941), Ceplene (histamine dihydrochloride), Orathecin (rubitecan), Virulizin, Gastrimmune (G17DT), DX-8951f (exatecan mesylate), Onconase (ranpirnase), BEC2 (mitumoab), Xcytrin (motexafin gadolinium) and combinations thereof.
  • anti-tumor agents may be selected from the following agents, CeaVac (CEA), NeuTrexin (trimetresate glucuronate) and combinations thereof. Additional antitumor agents may be selected from the following agents, OvaRex (oregovomab), Osidem (IDM-1), and combinations thereof. Additional anti-tumor agents may be selected from the following agents, Advexin (ING 201), Tirazone (tirapazamine), and combinations thereof. Additional anti-tumor agents may be selected from the following agents, RSR13 (efaproxiral), Cotara (131! chTNT 1/b), NBI-3001 (IL-4) and combinations thereof.
  • Additional anti-tumor agents may be selected from the following agents, Canvaxin, GMK vaccine, PEG lnteron A, Taxoprexin (DHA/paciltaxel), and combinations thereof.
  • Other anti-tumor agents include Pfizer's MEK1/2 inhibitor PD325901, Array
  • mTOR inhibitors may also be utilized such as CCI-779 (Wyeth) and rapamycin derivatives RAD001 (Novartis) and AP-23573 (Ariad), HDAC inhibitors, SAHA (Merck Inc./Aton Pharmaceuticals) and combinations thereof.
  • Additional anti-tumor agents include aurora 2 inhibitor VX-680 (Vertex), and Chk1/2 inhibitor XL844 (Exilixis).
  • cytotoxic agents e.g., one or more selected from the group consisting of epirubicin (Ellence), docetaxel (Taxotere), paclitaxel, Zinecard (dexrazoxane), rituximab (Rituxan) imatinib mesylate (Gleevec), and combinations thereof, may be used in combination with a compound of the present invention and pharmaceutical compositions disclosed herein.
  • the invention also contemplates the use of the compounds of the present invention together with hormonal therapy, including but not limited to, exemestane (Aromasin, Pfizer Inc.), leuprorelin (Lupron or Leuplin, TAP/Abbott/Takeda), anastrozole (Arimidex, Astrazeneca), gosrelin (Zoladex, AstraZeneca), doxercalciferol, fadrozole, formestane, tamoxifen citrate (tamoxifen, Nolvadex, AstraZeneca), Casodex (AstraZeneca), Abarelix (Praecis), Trelstar, and combinations thereof.
  • exemestane Amasin, Pfizer Inc.
  • leuprorelin Louprorelin
  • anastrozole Arimidex, Astrazeneca
  • gosrelin Zoladex, AstraZeneca
  • doxercalciferol
  • the invention also relates to the use of the compounds of the present invention together with hormonal therapy agents such as anti-estrogens including, but not limited to fulvestrant, toremifene, raloxifene, lasofoxifene, letrozole (Femara, Novartis), anti- androgens such as bicalutamide, flutamide, mifepristone, nilutamide, CasodexTM (4 1 - cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3'-(trifluoromethyl) propionanilide, bicalutamide) and combinations thereof.
  • hormonal therapy agents such as anti-estrogens including, but not limited to fulvestrant, toremifene, raloxifene, lasofoxifene, letrozole (Femara, Novartis), anti- androgens such as bicalutamide, flutamide, mife
  • the invention provides a compound of the present invention alone or in combination with one or more supportive care products, e.g., a product selected from the group consisting of Filgrastim (Neupogen), ondansetron (Zofran), Fragmin, Procrit, Aloxi, Emend, or combinations thereof.
  • supportive care products e.g., a product selected from the group consisting of Filgrastim (Neupogen), ondansetron (Zofran), Fragmin, Procrit, Aloxi, Emend, or combinations thereof.
  • Particularly preferred cytotoxic agents include Camptosar, Erbitux, Iressa, Gleevec, Taxotere and combinations thereof.
  • topoisomerase I inhibitors may be utilized as anti-tumor agents: camptothecin; irinotecan HCI (Camptosar); edotecarin; orathecin (Supergen); exatecan (Daiichi); BN-80915 (Roche); and combinations thereof.
  • Particularly preferred toposimerase Il inhibitors include epirubicin (Ellence).
  • Alkylating agents include, but are not limited to, nitrogen mustard N-oxide, cyclophosphamide, ifosfamide, melphalan, busulfan, mitobronitol, carboquone, thiotepa, ranimustine, nimustine, temozolomide, AMD-473, altretamine, AP-5280, apaziquone, brostallicin, bendamustine, carmustine, estramustine, fotemustine, glufosfamide, ifosfamide, KVV-2170, mafosfamide, and mitolactol; platinum-coordinated alkylating compounds include but are not limited to, cisplatin, Paraplatin (carboplatin), eptaplatin, lobapiatin, nedaplatin, Eloxatin (oxaliplatin, Sanofi) or sat ⁇ latin and combinations thereof. Particularly preferred alkylating agents include Eloxatin (ox
  • Antimetabolites include but are not limited to, methotrexate, 6-mercapt ⁇ purine riboside, mercaptopurine, 5-fluorouracil (5-FU) alone or in combination with leucovorin, tegafur, UFT, doxifluridine, carmofur, cytarabine, cytarabine ocfosfate, enocitabine, S-1 , Alimta (premetrexed disodium, LY231514, MTA), Gemzar (gemcitabine, EIi Lilly), fludarabin, 5-azacitidine, capecitabine, cladribine, clofarabine, decitabine, eflornithine, ethynylcytidine, cytosine arabinoside, hydroxyurea, TS-1 , melphalan, nelarabine, nolatrexed, ocfosfate, disodium premetrexed,
  • Antibiotics include intercalating antibiotics and include, but are not limited to: aclarubicin, actinomycin D, amrubicin, annamycin, adriamycin, bleomycin, daunorubicin, doxorubicin, elsamitrucin, epirubicin, galarubicin, idarubicin, mitomycin C, nemorubicin, neocarzinostatin, peplomycin, pirarubicin, rebeccamycin, stimalamer, streptozocin, valrubicin, zinostatin and combinations thereof.
  • Plant derived anti-tumor substances include for example those selected from mitotic inhibitors, for example vinblastine, docetaxel (Taxotere), paclitaxel and combinations thereof.
  • Cytotoxic topoisomerase inhibiting agents include one or more agents selected from the group consisting of aclarubicn, amonafide, belotecan, camptothecin, 10- hydroxycamptothecin, 9-aminocamptothecin, diflomotecan, irinotecan HCI (Camptosar), edotecarin, epirubicin (Ellence), etoposide, exatecan, gimatecan, lurtotecan, mitoxantrone, pirarubicin, pixantrone, rubitecan, sobuzoxane, SN-38, tafluposide, topotecan, and combinations thereof.
  • Preferred cytotoxic topoisomerase inhibiting agents include one or more agents selected from the group consisting of camptothecin, 10-hydroxycamptothecin, 9- aminocamptothecin, irinotecan HCI (Camptosar), edotecarin, epirubicin (Ellence), etoposide, SN-38, topotecan, and combinations thereof.
  • Interferons include interferon alpha, interferon alpha ⁇ 2a, interferon, alpha-2b, interferon beta, interferon gamma-1a, interferon gamma-1b (Actimmune), or interferon gamma-n1 and combinations thereof.
  • agents include filgrastim, lentinan, sizofilan, TheraCys, ubenimex, WF-10, aldesleukin, alemtuzumab, BAM-002, dacarbazine, daclizumab, denileukin, gemtuzumab ozogamicin, ibritumomab, imiquimod, lenograstim, lentinan, melanoma vaccine (Corixa), molgramostim, OncoVAX-CL, sargramostim, tasonermin, tecleukin, thymalasin, tositumomab, Virulizin, Z-100, epratuzumab, mit ⁇ momab, oregovomab, pemtumomab (Y-muHMFG1), Provenge (Dendreon) and combinations thereof.
  • Biological response modifiers are agents that modify defense mechanisms of living organisms or biological responses, such as survival, growth, or differentiation of tissue cells to direct them to have anti-tumor activity.
  • agents include krestin, lentinan, sizofiran, picibanil, ubenimex and combinations thereof.
  • Other anticancer agents that can be used in combination with a compound of the present invention include alitretinoin, ampligen, atrasentan bexarotene, bortezomib.
  • Bosentan calcitriol, exisulind, finasteride.fotemustine, ibandronic acid, miltefosine, mitoxantrone, l-asparaginase, procarbazine, dacarbazine, hydroxycarbamide, pegaspargase, pentostatin, tazarotne, Telcyta (TLK-286, Teiik Inc.), Velcade (bortemazib, Millenium), tretinoin, and combinations thereof.
  • Platinum-coordinated compounds include but are not limited to, cisplatin, carboplatin, nedaplatin, oxaliplatin, and combinations thereof.
  • Camptothecin derivatives include but are not limited to camptothecin, 10- hydroxycamptothecin, 9-aminocamptothecin, irinotecan, SN-38, edotecarin, topotecan and combinations thereof.
  • antitumor agents include mitoxantrone, l-asparaginase, procarbazine, dacarbazine, hydroxycarbamide, pentostatin, tretinoin and combinations thereof.
  • Anti-tumor agents capable of enhancing antitumor immune responses such as CTLA4 (cytotoxic lymphocyte antigen 4) antibodies, and other agents capable of blocking CTLA4 may also be utilized, such as MDX-010 (Medarex) and CTLA4 compounds disclosed in U.S. Patent 6,682,736; and antiproliferative agents such as other famesyl protein transferase inhibitors, for example the farnesyl protein transferase inhibitors.
  • CTLA4 antibodies that can be used in combination with compounds of the present invention include those disclosed in U.S. Patents 6,682,736 and 6,682,736 both of which are herein incorporated by reference in their entirety.
  • Specific IGF1R antibodies that can be used in the combination methods of the present invention include those disclosed in WO 02/053596, which is herein incorporated by reference in its entirety.
  • Gene therapy agents may also be employed as anti-tumor agents such as TNFerade (GeneVec), which express TNFalpha in response to radiotherapy.
  • statins may be used in combination with a compound of the present invention and pharmaceutical compositions thereof.
  • Statins HMG-CoA reducatase inhibitors
  • Atorvastatin LipitorTM, Pfizer Inc.
  • Pravastatin Pravastatin
  • Lovastatin MevacorTM, Merck Inc.
  • Simvastatin ZaocorTM, Merck Inc.
  • Fluvastatin LescolTM, Novartis
  • Cerivastatin BoycolTM, Bayer
  • Rosuvastatin CrestorTM, AstraZeneca
  • Lovostatin and Niacin Niacin
  • statin is selected from the group consisting of Atovorstatin and Lovastatin, derivatives and combinations thereof.
  • Other agents useful as anti-tumor agents include Caduet.
  • non-exemplified compounds according to the invention may be performed by modifications apparent to those skilled in the art, e.g., by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by making routine modifications of reaction conditions.
  • other reactions disclosed herein or known in the art will be recognized as having adaptability for preparing other compounds of the invention.
  • DMF means di-methyl formamide
  • DMAP means di-methyl amino pyridine
  • TME tert-butyl methyl ether
  • Me means methyl
  • TAA tri-ethyl amine
  • i-PrOH means isopropyl alcohol
  • HATU means O-(7-azabenzotriazol-1-yl)-N,N,N',N ) -tetramethyluronium phosphorus pentafluoride
  • DMSO means di-methyl sulfoxide
  • EtOAc means ethyl acetate
  • Et means ethyl
  • Ph means phenyl
  • Boc means t-butyloxycarbonyl
  • DCM means di-chloro methane
  • DME means di-methyl ether
  • MeOH means methanol
  • THF means tetrahydrofuran
  • DCM means di-chloro methane
  • DME means di-methyl ether
  • MeOH means
  • R 1 , R 2 , R 3 halogen, optionally substituted alkyl, optionally substituted alkoxy, amino, carboxyamid ⁇ , cyano, aryl or heteroaryl.
  • R 1 , R 2 , R 3 halogen, optionally substituted alkyl, optionally substituted alkoxy, amino, carboxyamide, cyano, aryl, or heteroaryl.
  • R H, alkoxyl, amino, alkylcyano, fluoroalkyl, optionally substituted (C 1 -C ⁇ aHCyI 1 fluoroalkyl, optionally substituted aryl or optionally substituted heteroaryl
  • R 1 , R 2 , R 3 halogen, optionally substitued alkyl, optionally substituted alkoxy, amino, carboxyamide, cyano, aryl or heteroaryl.
  • BF 3 OEt 2 Into a 2 L 3-neck flask was added BF 3 OEt 2 (70.6 mL, 79.7 g, 560 mmol) and the system was cooled to O 0 C. To this system was added a solution of 5-amino-2,4-dichlorophenol (50.0 g, 280 mmol) in 700 mL of THF over 45 minutes. Upon completion of addition, a solution of isoamyl nitrite (48.8 mL, 42.78 g, 365 mmol) in 150 mL of THF was added over 15 minutes. The reaction was stirred an additional 30 minutes at O 0 C during which time a yellow precipitate formed.
  • 5-amino-2,4-dichlorophenol 50.0 g, 280 mmol
  • isoamyl nitrite 48.8 mL, 42.78 g, 365 mmol
  • Et 2 O was added to increase precipitation and the yellow solid was filtered, washed with Et 2 O, and air dried.
  • the precipitate was then added to a solution of NaI (54.7 g, 365 mmol) in 1.2 L of acetone portionwise and the red solution was stirred at ambient temperature overnight.
  • Solvent was removed by rotary evaporation leaving an orange semisolid. Water was added and the mixture was extracted 2x with EtOAc. The combined organics were washed with saturated NaHS ⁇ 3 and dried over Na 2 SO 4 . After filtration of the drying agent, solvent was removed in vacuo leaving an orange oil.
  • R alkoxyl, amino, alkylcyano, fluoroalkyl, optionally substituted (C,-C 6 )a!kyl, optionally substituted aryl or optionally substituted heteroaryl
  • R' H, alkyl, cyano, amino, fluoroalkyl, carboxyamide, or carboxylic acid
  • Examples 1 to 13 provide detailed synthetic steps for preparing several specific compounds of the present invention.
  • Table 1 shows additional compounds that were prepared as Examples 14 to 15 according to the general reaction schemes as described herein.
  • Examples 16 and 17 describe the biochemical and cellular assays used to assess the potency of the compounds shown in Examples 1 to 15.
  • Table 3 shows the biochemical and celluar assay values for compounds prepared as Examples 1 to 15.
  • the compounds shown in the examples below may have asymmetric carbon atoms.
  • the carbon-carbon bonds of the compounds of the present invention may be depicted herein using a solid line ( ), a solid wedge ( -" ), or a dotted wedge ( "" ).
  • a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that the compound was prepared as a racemic mixture, or that the compound was prepared as a pure enantiomer but the absolute stereochemistry was not determined.
  • the use of either a solid or dotted wedge to depict bonds to asymmetric carbon atoms is meant to indicate that the compound was prepared as a stereoisomer where the absolute stereochemistry was determined.
  • [1 ,3,2]dioxaborolane (compound A in General Procedure GD, 362 mg, 1 mmo!) was added to a solution of N'-(4-Chloro-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-N 1 N-dimethyl- formamidine (compound 2 in General Procedure GB2, 233 mg, 1 mmol) in 7 ml_ of 1 ,4- dioxane. The mixture was purged with N 2 several times. Tetrakis (triphenylphosphine) palladium (115 mg, 0.1 mmol) was added, then Na 2 COa (1 ml_, 2M) was added to the mixture.
  • Tetrakis (triphenylphosphine) palladium (115 mg, 0.1 mmol) was added, then Na 2 COa (1 mL, 2M) was added to the mixture. The mixture was heated and stirred at 85 0 C for 12 hours. H 2 O (20 mL) was added to the reaction mixture. EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil as the desired product.
  • Tetrakis (triphenylphosphine) palladium 120 mg, 0.1 mmol was added then Na 2 COa (1.5 mL, 2M) was added to the mixture. The mixture was heated and stirred at 85 0 C for 12 hours. H 2 O (30 mL) was added to the reaction mixture. EtOAc (2x 100 mL) was added to extract the aqueous solution.
  • [1,3,2]dioxaborolane (compound E in General Procedure GD, 120 mg, 0.43 mmol) was added to a solution of N'-(4-Chloro-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-N,N-dimethyl- formamidine (compound 2 in General Procedure GB2, 96.5 mg, 0.43 mmol) in 6 mL of 1 ,4-dioxane. The mixture was purged with N 2 several times. Tetrakis (triphenylphosphine) palladium (50 mg, 0.043 mmol) was added, then Na 2 CO 3 (0.65 ml_, 2M) was added to the mixture.
  • Example 8 4- ⁇ 2,4-dichloro-5-[2-(diethylamino)ethoxy]phenyl ⁇ -5H-pyrrolo[3,2- d]pyrimidin-2-amine
  • the above compound was prepared as follows using General Procedure GB3 as described previously. Sodium hydride (15 mg, 0.4 mmol) was added to a solution of the final compound in Example 2 (41 mg, 0.14 mmol) in DMF. The mixture was stirred at room temperature for 30 minutes. Potassium carbonate (40 mg, 0.3 mmol) was added as a solid and then (2-Bromo-ethyl)-diethyl-amine hydrobromide (60 mg, 0.3 mmol) was added. The reaction was heated at 8O 0 C for 12 hours. H 2 O (50 mL) was added, EtOAc (50 mL) and THF (50 mL) were added to extract the aqueous solution.
  • Example 12 7-(4-bromo-2-chloro-5-methoxyphenyl)-2-methyl[1 ,3]thiazolo[5,4- d]pyrimidin-5-amine
  • Example 16 7-[2,4-Dichloro-6-(2-pyrazol-1 -yl-ethoxy)-phenyl]-thiazolo[5,4- d]pyrimidin-5-ylamine.
  • Ethyl cyanoglyoxylate-2-oxime 50 g, 0.35 mol was suspended in H 2 O (300 ml). Sat. aq. NaHC ⁇ 3 solution (150 ml) added in portions to the suspension and the suspension became a orange solution. Sodium dithionite (85% tech, 170 g, 0.98 mol) was scooped in portions. After the addition the mixture was stirred for 30 minutes and the temperature rose to 42 0 C. The reaction mixture was them immediately extracted with CH 3 CI (4x 200 ml). The combined organic fraction was dried (Na2SO 4 ), filtered and concentrated under reduced pressure affording a red/brown oil (34 g, 265 mmol, 76%).
  • Benzoyl isothiocyanate (0.4ml, 3.02 mmol) was added to a solution of compound 16c (500 mg, 2.9 mmol) in dioxane (25 mL) at room temperature under nitrogen. The mixture was heated to reflux (about 100 0 C) for 12h. The reaction was monitored by LC/MS. The solvent was evaporated to afford a white solid residue, which was triturated with heptanes. The colorless solid was collected by filtration to afford compound 16d (830 mg, 2.48 mmol, 85% yield).
  • lodomethane (43.3 mg, 0.31 mmol) was added to a solution of the compound of Example 17 (100 mg, 0.22 mol) and K 2 CO 3 (151 mg, 1.1 mmol) in DMF (5 mL). The mixture was stirred at room temperature for 12h and monitored by LC/MS. The mixture was acidified with HOAc-NaOAc buffer and EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a yellow oil.
  • Example 19 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-5-methoxythieno[2,3- d]pyrimidine-6-carboxylic acid.
  • Lithium hydroxide (207 mg, 4.93 mmol) in H 2 O (3M, 1.6 mL) was added to a solution of compound of Example 18 (233 mg, 0.49 mmol) in dioxane (5 mL). The mixture was heated at 60 0 C for 12h. The reaction was monitored by LC/MS. The mixture was acidified with HOAc-NaOAc buffer and EtOAc (2x100 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to afford the compound of Example 19 as a yellow solid (166 mg, 75.7% yield).
  • Example 19 (56 mg, 0.13 mmole), diisopropylethylamine (100 mg, 0.8 mmol, 0.13 mL), and O-(7-azabenzotriazol-1-yl)-N,N ) N',N'-tetramethyluronium phosphorus pentafloride (HATU) (53 mg, 0.14 mmol) in DMF (5 mL) under a nitrogen atmosphere. The reaction was allowed to stir at room temperature for 12 h. Saturated NaHCCb was added to the reaction mixture to quench the reaction. EtOAc (2x50 mL) was then added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a brown yellow oil.
  • HATU O-(7-azabenzotriazol-1-yl)-N,N ) N',N'-tetramethyluronium phosphorus pentafloride
  • Example 21 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-N-ethyl-5- hydroxythieno[2,3-d]pyrimidine-6-carboxamide.
  • Lithium hydroxide (317 mg, 2.5 mL, 3 M in H2O) was added to a solution of 4-(4-Bromo- 2-chloro-5-methoxy-phenyl)-5-ethoxymethoxy-2-methylsulfanyl-thieno[2,3-d]pyrimidine- 6-carboxylic acid ethyl ester in dioxane (5 mL). The mixture was heated at 60 0 C for 12h and monitored by LC/MS. The reaction was neutralized with HOAc-NaOAc buffer and extracted with EtOAc (2x100 mL). The combined organic layer was dried, filtered, and concentrated to get a brown foam as the desired product (328 mg, 83.5% yield).
  • Ethylamine (198 mg, 4.4 mmol) was added to a solution of 4-(4-Bromo-2-chloro-5- methoxy-phenyl)-5-ethoxymethoxy-2-methylsulfanyl-thieno[2,3-d]pyrimidine-6-carboxylic acid (228 mg, 0.44 mmole), diisopropylethylamine (340 mg, 2.6 mmol, 0.5 mL), and O- (7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium phosphorus pentafloride (HATU) (250 mg, 0.66 mmol) in DMF (7 mL) under a nitrogen atmosphere.
  • HATU O- (7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium phosphorus pentafloride
  • Example 22 N-((1 R,5S)-3-aza-bicyclo[3.1.0]hexan-6-yl)-2-amino-4-(2,4-dimethoxy- 6-methylphenyl)thieno[2,3-d]pyrimidine-6- carboxamide.
  • Lithium hydroxide (320 mg, 7.5 mmol, 20 equiv., 2 M in H 2 O) was added to a solution of compound 22b in dioxane (5 mL). The mixture was stirred at 60 °C for 12h. The mixture was neutralized with HOAc-NaOAc buffer and extracted with EtOAc (2x50 mL). The combined organic layer was dried, filtered, and concentrated to get a pale yellow solid as the desired product. This compound was used for the next step reaction without further purification.
  • Example 23 ethyl 4-(2-(2-(1H-pyrazol-1-yl)ethoxy)-4,6-dichlorophenyl)-2- aminothieno[2,3-d]pyrimidine-6-carboxylate.
  • Compound 23a was made following the same method that compound 22b was made.
  • HSP-90 Compounds of the present invention were evaluated for potency against HSP-90 using a SPA (scintillation proximity assay) competition binding assay. Briefly, either full length or N-terminal HSP-90 that contains a 6-His tag on its C-terminus binds to copper on Yttrium-silicate scintillant beads via the His-tag. Tritiated propyl-Geldanamycin (pGA), whose structure is shown below, is an analog of a natural inhibitor of HSP-90 called Geldanamycin.
  • pGA Tritiated propyl-Geldanamycin
  • Tritiated pGA which contains a tritiated propyl-amine group added at the #17 position, binds HSP-90 and brings the isotope into proximity with the beads.
  • 17-n-propylamino- Geldanamycin can be prepared as described in U.S. Patent No. 4,261 ,989, which is incorporated herein by reference.
  • a second tritiated compound that can also be used in this assay is shown below and is designated as Compound A.
  • the "T” in the structure above indicates the position of the labeled tritiated hydrogen atoms.
  • This compound has a K d of 40 nM and can be prepared as follows.
  • the parent compound (/V-allyl-2-(5-chloro-2,4-dihydroxybenzoyl)isoindoline-1- carboxamide) shown above was first prepared as follows. Allylamine (2.5 ml_, 5 mmol, 2M in THF) was added to a solution of Boc(R,S)-1,3-dihydro-2H-isoindole carboxylic acid (263 mg, 1 mmole), diisopropylethy!
  • N-allylisoindoline-1-carboxamide (1 mmol) was then added to a solution of 5- chloro-2,4-bis(methoxymethoxy)benzoic acid (which can be prepared as shown in WO 2006/117669) (340 mg, 1.2 mmol), 4-methylmorpholine (2.2 mL, 20 mmol), N-(3- dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (460 mg, 2.4 mmol), and 1- hydroxy benzotriazole (330 mg, 2.4 mmol) in 12 mL of DMF under a nitrogen atmosphere. The reaction was allowed to stir at room temperature for 12 hours.
  • Compound A was prepared using standard hydrogenation methods using tritium gas.
  • the beta signal emitted from the isotope excites the scintillant, which creates a measurable signal.
  • competitive compounds are added to the assay mixture, they compete with bound tritiated pGA or Compound A at the ATP-binding site on the N- terminal of HSP-90.
  • the signal is reduced (the beta-particles are no longer in proximity with the bead). This reduction in signal is used to quantify the extent to which the inhibitor/compound is competitive with pGA or Compound A.
  • the SPA assay for 3 H-pGA (designated G1) and Compound A (designated G2) binding to HSP-90 was performed in 96-well flat bottom white plates (Corning #3604).
  • typical reaction solutions contained 30 nM HSP-90 and 200 nM 3 H-pGA in binding buffer (100 mM Hepes, pH 7.5 and 150 mM KCI).
  • typical reaction solutions contained 5 nM HSP-90 and 50 nM of Compound A.
  • the 3 H-pGA was first diluted to 33% label with unlabeled pGA that was synthesized and purified to give a final concentration of 200 nM.
  • labeled Compound A was diluted with unlabeled Compound A to provide a ratio of labeledrunlabeled of 1 :2 for a final concentration of 50 nM.
  • Inhibitors were added to the HSP-90/ 3 H-pGA (or HSP-90/Compound A) solutions at eleven different concentrations for Ki determinations. The range of inhibitor concentrations were 100 ⁇ M, or an appropriate range, for solid samples and 10 ⁇ M for targeted library compounds and 4 mM liquid stocks. To determine percent inhibition, the compound was tested at 1 and 10 ⁇ M. The final DMSO in the samples was 4%.
  • Copper-Ysi beads (Amersham, #RPNQ0096) that have been diluted in binding buffer were added to each well to give a final concentration of 100 ⁇ g/well.
  • the plates were sealed, covered with a foil-covered lid and shaken for 30 minutes at room temperature.
  • the beads were allowed to settle for 30 minutes after which the plates were counted using a Packard TopCount NXT instrument. This procedure has also been adapted for medium throughput using a Beckman Biomek FX. Samples were run in duplicate and on two separate days to assure an accurate value of Kj. For Ki determinations, the corrected cpm's (actual cpm's minus background) were plotted vs. inhibitor concentration using GraphPad Prism software.
  • IC50 is about the same as the HSP-90 concentration.
  • HSP-90 concentration For a tight binding inhibitor, the following equation can be applied:
  • EL and EL 0 are the radioligand-HSP-90 complexes in the presence and absence of inhibitor, respectively.
  • EUEL 0 represents the fractional signal in the presence of inhibitor, lo, E 0 , and L 0 are the inhibitor, HSP-90, and radioligand concentrations, respectively.
  • Ki is the inhibition constant for the ligand, while K L is the binding affinity constant between the enzyme (HSP-90) and the ligand.
  • Akt Luminex assay measures the turnover of the HSP- 90 client protein Akt1.
  • NCI-H1299 cells ATCC Number: CRL-5803
  • Cell lysates were then assayed to measure the loss of Akt1 expression and thus determine cellular IC 50 by Akt/PKB Beadmate (Upstate Catalog # 46-605) using a Luminex 100 system.
  • Table 2 HSP-90 Biochemical and cellular assay data for compounds shown in Examples 1 to 46.

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Abstract

The present invention is directed to 2-aminopyrimidine compounds and pharmaceutically acceptable salts thereof, their synthesis and their use as HSP-90 inhibitors, for the treatment of cancer.

Description

2-AMINO PYRIMIDINE COMPOUNDS
This application claims the benefit of United States Provisional Patent Application Nos. 60/866,373, filed November 17, 2006 and 60/984,938 filed November 2, 2007 the disclosures of which are incorporated herein by reference in their entireties.
Field of the Invention
The present invention is directed to compounds, and pharmaceutically acceptable salts and solvates thereof, their synthesis, and their use as modulators or inhibitors of HSP-90. The compounds of the present invention are useful for modulating (e.g. inhibiting) HSP-90 activity and for treating diseases or conditions mediated by HSP-90, such as for example, disease states associated with abnormal cell growth such as cancer.
Background Molecular chaperones play important roles in cellular function by ensuring proper folding of proteins upon synthesis as well as their refolding under conditions of denaturing stress. By regulating the balance between protein synthesis and degradation, molecular chaperones are a significant part of the cellular response to stress. In addition, by regulating the proper folding of various cellular proteins, chaperones play an important role in regulating cellular functions such as cell proliferation and apoptosis. (See, e.g. Jolly, et al., J. Natl. Cancer Inst. 92: 1564-1572 (2000)). Heat shock proteins (HSPs) are a class of chaperones that accumulate in the cell in response to various environmental stresses, such as heat shock, oxidative stress, or the presence of alcohols or heavy metals. In addition to their role in protecting the cell from such environmental stresses, HSPs may also play a significant role as chaperones for a variety of cellular proteins under stress-free conditions. Members of the HSP family are classified according to their molecular weight (e.g. HSP-27, HSP-70, and HSP-90). Evidence of differential expression of HSPs in various stages of tumor progression suggests HSPs play a role in cancer. (See, e.g. Martin, et al., Cancer Res. 60:2232-2238 (2000)). HSP-90 is a homodimer with ATPase activity and functions in a series of complex interactions with a variety of substrate proteins (Young, et al., J. Cell Biol. 154: 267-273 (2001)). HSP-90 is unique with regard to other chaperones, however, since most of its known substrate proteins are signal transduction proteins. Thus, HSP-90 plays an essential role in regulating cellular signal transduction networks. (See, e.g. Xu, et al., Proc. Natl. Acad. Sci 90:7074-7078 (1993)). In particular, substrate proteins of HSP-90 include many mutated or over-expressed proteins implicated in cancer such as p53, Bcr-At>1 kinase, Raf-1 kinase, Akt kinase, Npm-Alk kinase p185ErbB2 transmembrane kinase, Cdk4, Cdk6, Wee1 (a cell cycle-dependent kinase), HER2/Neu (ErbB2), and hypoxia inducible factor-l et (HIF-Ia). Thus inhibition of HSP-90 results in selective degradation of these important signaling proteins involved in apoptosis, cell proliferation, and cell cycle regulation (Holstein, et al., Cancer Res. 61 :4003-4009 (2001)). Accordingly, HSP-90 is an attractive therapeutic target because of the important roles played by these signaling proteins in disease states involving abnormal cell growth, such as cancer. It is thus desirable to discover and develop new inhibitors of HSP-90 activity that can provide a therapeutic benefit to patients suffering from disease states related to abnormal cell growth such as cancer. Examples of compounds that inhibit HSP-90 include those described in published patent applications WO 2006/079789, WO 2006/008503, WO 2006/090094, and WO 2005/021552.
Summary The present invention provides compounds of formula (I)
Figure imgf000003_0001
(I) wherein:
X is CR2, N, or NR2;
Y is CH or S, provided that when X is CR2, Y is S;
R1 is -(CHa)n-(C6 to C14) aryl, -(CH2)n-(C2 to C9) heteroaryl, or -(CH2)n-C(O)NR4aR4b, wherein each of said (C6 to Cu) aryl and (C2 to Cg) heteroaryl is optionally substituted with at least one R5 group; R2 is H, -(CHb)n-(Ce to Ci4) aryl, -(CH2)n-(C2 to C9) heteroaryl, or - (CH2)n-C(O)NR4aR4b, wherein each of said (C6 to C14) aryl and (C2 to C9) heteroaryl is optionally substituted with at least one R5 group;
R3 is H, (Ci to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, -(CH2)n-C(O)OR7 or -(CH2)n-C(O)NR4aR4b;
R4a and R4b are each independently H, (Ci to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, (C6 to Cu) aryl, (C2 to C9) heteroaryl, (C2 to C9) cycloheteroalkyl, or (C3 to C8) cycloalkyl, all optionally substituted with at least one R8 group, or R4a and R4b, taken together with the nitrogen atom to which they are bound, form a (C2 to C9) cycloheteroalkyl group, wherein said (C2 to C9) cycloheteroalkyl is optionally substituted with at least one R8 group; each R5 is independently -OH, (C1 to C6) alkyl, cyano, halogen, -C(O)NR4aR4b, -NR4aR4b, (C1 to C8) alkoxy, (C6 to C14) aryl or (C2 to C9) heteroaryl wherein each of said (Ci to C8) alkoxy, (C6 to Cu) aryl, and (C2 to C9) heteroaryl is optionally substituted with at least one R6 group; each R6 is independently halogen, (C-i to C6) alkyl, (C6 to C14) aryl, (C3 to C8) cycloalkyl, (C2 to C9) heteroaryl, (C2 to C9) cycloheteroalkyl, -(CH2)n-C(O)NR4aR4b, or -NR4aR4b;
R7 is H, or (C1 to C6) alkyl; each R8 is independently -OH, (C1 to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, (C1 to C8) heteroalkyl, or (C1 to C8) alkoxy; each n is independently 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof.
In one aspect of the present invention, the compound of formula (I) has the following structure:
Figure imgf000004_0001
In one embodiment, R1 is (Ce to Ci4) aryl or (C2 to Cg) heteroaryl, wherein each of said (Cβ to Cu) aryl and (C2 to C9) heteroaryl is optionally substituted with at least one R5 group.
In a further embodiment, R1 is Cβ aryl, wherein said Ce aryl is optionally substituted with at least one R5 group.
In a further embodiment, R3 is H or -CH3. In another embodiment, R3 is -(CH2)n-C(O)NR4aR4b. In another embodiment, R3 is -C(O)NHR4b.
In a further aspect of the present invention, the compound of formula (I) has the following structure:
Figure imgf000005_0001
wherein:
R5a is (Ci to C8) alkoxy, which is optionally substituted with at least one R6 group; R5b is halogen; and R5G is halogen. In one particular embodiment, the invention relates to the compound or salt shown above, wherein R5a is -OCH3, R5b is Br, and R50 is Cl.
In a further embodiment, the compound of formula (I) has the following structure:
Figure imgf000005_0002
wherein:
R is (Ci to C8) alkoxy, which is optionally substituted with at least one R group;
R5b is halogen; and F 3r 5c3 is halogen.
In one particular embodiment, the invention relates to the compound or salt shown above, wherein R5a is -OCH3, R5b is Br, and R5c is Cl.
In a further embodiment of the present invention the compound of formula (I) has the following structure:
Figure imgf000006_0001
In one particular embodiment, the invention relates to the compound or salt described above, wherein R1 is (Cβ to Ci4) aryl or (C-2 to C9) heteroaryl, wherein each of said (Ce to Cu) aryl and (C2 to C9) heteroaryl is optionally substituted with at least one R5 group.
In a further embodiment, R1 is Ce aryl, wherein said Ce aryl is optionally substituted with at least one R5 group. In a further embodiment, R3 is H or -CH3. In a further embodiment, R3 is -(CH2)n-C(O)NR4aR4b. In a further embodiment, R3 is - C(O)NHR4b.
In a further embodiment of the present invention the compound or salt described above has the following structure:
Figure imgf000006_0002
wherein:
R5a is (C1 to C8) alkoxy, which is optionally substituted with at least one R6 group;
R5b is halogen; and
R5c is halogen.
In one particular embodiment, R5a is -OCH3, R5b is Br, and R5c is Cl. In a further embodiment of the present invention, the compound or salt described above has the following structure:
Figure imgf000007_0001
wherein: R5a is (Ci to Cs) alkoxy, which is optionally substituted with at least one R6 group;
R5b is halogen; and R5c is halogen.
In one particular embodiment, R5a is -OCH3, R5b is Br, and R5c is Cl. In a further aspect of the present invention is a compound as described previously, wherein the compound of formula (I) has the following structure:
Figure imgf000007_0002
In one particular embodiment, R1 is (C6 to Cu) aryl or (C2 to C9) heteroaryl, wherein each of said (C6 to Ci4) aryl and (C2 to C9) heteroaryl is optionally substituted with at least one R5 group.
In a further embodiment, R1 is C6 aryl, wherein said C6 aryl is optionally substituted with at least one R5 group.
In a further embodiment, R3 is H or -CH3. In another embodiment, R3 is -(CH2)n-C(O)NR4aR4b. In another embodiment, R3 is -C(O)NHR4b.
In a further embodiment, the compound of formula (I) has the following structure:
Figure imgf000008_0001
wherein:
R5a is (Ci to Cs) alkoxy, which is optionally substituted with at least one R6 group;
R5b is halogen; and
R50 is halogen.
In one particular embodiment, R5a is -OCH3, R5b is Br, and R5c is Cl.
In a further embodiment, the compound of formula (I) has the following structure:
Figure imgf000008_0002
wherein: R5a is (Ci to Ce) alkoxy, which is optionally substituted with at least one R6 group;
R5b is halogen; and
RSc is halogen.
In one particular embodiment, R5a is -OCH3, R5b is Br, and R5° is Cl.
It should be understood that the present invention encompasses compounds of formula (I) as described above formed by any and all combinations of the definitions of
R1, R2, R3, R4a, R4b, R5, R5a, R5b, R50, R6, R7, R8, and n as described above, and further includes pharmaceutically acceptable salts and solvates thereof of any of said compounds.
The present invention also provides a compound of formula (II),
Figure imgf000009_0001
(H) wherein:
X is CR9 or N; R9 is H, -OH, halogen, (Ci to C6) alkyl, (Ci to C6) alkoxy or (Ci to C6) perfluoroalkyl;
R10 is selected from the group consisting of
(a) H, (Ci to C6) perfluoroalkyl, (Ci to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl and (Ci to C6) alkoxy, (b) (C1 to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl and (Ci to C6) alkoxy, all further substituted by 1-3 groups selected from halogen, -OH, cyano, -NH2, -NH-(Ci to C3 alkyl), -N(Ci to C3 alkyl)2, (C3 to Ca) cycloalkyl, (C2 to C9) cycloheteroalkyl, phenyl and (C2 to C9) heteroaryl, wherein each (C3 to Ce) cycloalkyl, (C2 to Cg) cycloheteroalkyl, phenyl and (C2 to Cg) heteroaryl is optionally further substituted by 1-3 groups selected from halogen and (Ci to C3 alkyl),
(c) (C3 to Cs cycloalkyl), (C2 to C9) cycloheteroalkyl, phenyl and (C2 to Cg) heteroaryl, all optionally further substitued by 1-3 groups selected from (Ci to C6) alkyl, halogen, -OH, cyano, (Ci to C6) alkoxy, -S(O)2-(C1 to C3 alkyl), -NH2, -NH-(Ci to C3 alkyl) and -N(Ci to C3 alkyl)2, and (d) -(CH2)P-C(O)R12,
(e) -(CH2)P-C(O)-OR12, and
(f) -(CH2)p-C(O)NR13aR13b; each R11a , R11b and R11c is independently selected from the group consisting of
(a) -OH, halogen, cyano, (C-i to C6) perfluoroalkyl, (Ci to C6) alkyl, (C2 to C6) alkenyl and (C2 to C6) alkynyl,
(b) (C1 to C6) alkyl, (C2 to C6) alkenyl and (C2 to C6) alkynyl, all further substituted by 1-3 groups selected from halogen, -OH, cyano, -S(O)2-(Ci to C3 alkyl), -NH2, -NH- (Ci to C3 alkyl), -N(Ci to C3 alkyl)2, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, phenyl and (C2 to Cg) heteroaryl, wherein each (C3 to C8) cycloalkyl, (C2 to Cg) cycloheteroalkyl, phenyl and (C2 to C9) heteroaryl is optionally further substituted by 1-3 groups selected from halogen and (Ci to C3 alkyl), (c) (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, all optionally further subsitued by 1-3 groups selected from (Ci to C6) alkyl, halogen, -OH, cyano, -S(O)2-(Ci to C3 alkyl), (C1 to C6) alkoxy, -NH2, -NH-(C1 to C3 alkyl) and -N(C1 to C3 alkyl)2,
(d) -C(O)-R15, -(C-, to C6 alkylene)-C(O)R15, -(C2 to C6 alkenylene)-C(O)R15 and - (C2 to C6 alkynylene)-C(O)R15, (e) -0-R15, -0-(C1 to C6 alkylene)-R15, -0-(C2 to C6 alkenylene)-R15 and -0-(C2 to
C6 alkynylene)-R15,
(f) -0-(C1 to C6 alkylene)-C(0)0R15, -0-(C2 to C6 alkenylene)-C(O)OR15 and -O- (C2 to C6 alkynyleπe)-C(O)OR15;
(g) -O-(Ci to C6 alkylene)-S(O)2R15, -0-(C2 to C6 alkenylene)-S(O)2R15 and -0-(C2 to C6 alkynylene)-S(O)2R15;
(h) -0-(C1 to C6 alkylene)-C(O)NR1δaR16b, -0-(C2 to C6 alkenylene)- C(O)NR16aR16b and -0-(C2 to C6 alkynylene)-C(O) NR16aR16b, and
(i) -0-(C1 to C6 alkylene)-NR16aR16b, -0-(C2 to C6 alkenylene)-NR16aR16b and -O- (C2 to C6 alkynylene)-NR16aR16b; each R12 is independently H, (C1 to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl,
(C1 to C6) perfluoroalkyl, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, (C6 to Cu) aryl, (C2 to Cg) heteroaryl, -(Ci to C3 alkylene)-(C3 to Ce) cyclcoalkyl, -(Ci to C3 alkylene)- (C2 to C9 cycloheteroalkyl), -(Ci to C3 alkylene)- (C6 to C14 aryl) or -(C1 to C3 alkylene)- (C2 to C9 heteroaryl), wherein each R12 is optionally further substituted by 1-3 R14; each R13a and R13b is H, (C1 to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl, (C1 to C6) perfluoroalkyl, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, (C6 to C14)aryl, (C2 to C9) heteroaryl, -(C1 to C3 alkylene)-(C3 to Cs) cyclcoalkyl, -(Ci to C3 alkylene)- (C2 to C9 cycloheteroalkyl), -(Ci to C3 alkylene)- (C6 to Cu aryl) or -(Ci to C3 alkylene)- (C2 to C9 heteroaryl), each R13a or R13b is optionally further substituted by 1-3 R14; or, R13a and R13b, taken together with the nitrogen atom to which they are bound, form a (C2 to C9) cycloheteroalkyl group or a (C2 to C9) heteroaryl group, wherein said (C2 to C9) cycloheteroalkyl or (C2 to Cg) heteroaryl group is optionally further substituted by 1-3 R14; each R14 is independent halogen, -OH, cyano, -S(O)2-(Ci to C3 alkyl), (Ci to C3) alkyl, (Ci to C3) perfluoroalkyl (Ci to C3) alkoxy, -C(O)-(CrC6 alkyl), -C(O)-O(Ci to C6 alkyl), -C(O)-NH2, -C(O)-NH(Ci to C6 alkyl) or -C(O)N(Ci to C6 alkyl)2; each R15 is independently H, (Ci to C6) alkyl, (C2 to C6) alkenyl, (C2 to Ce) alkynyl,
(Ci to C6) perfluoroalkyl, (C3 to Ce) cycloalkyl, (C2 to C9) cycloheteroalkyl, (C6 to Cu)aryl,
(C2 to Cg) heteroaryl, -(Ci to C3 alkylene)-(C3 to Cs) cyclcoalkyl, -(Ci to C3 alkylene)- (C2 to Cg cycloheteroalkyl), -(Ci to C3 alkylene)- (C6 to Cu aryl) or -(Ci to C3 alkylene)- (C2 to Cg heteroaryl), wherein each R15 is otpioanlly further substituted by 1-3 R17; each R16a and R16b is independently H, (Ci to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl, (Ci to C6) perfluoroalkyl, (C3 to Ca) cycloalkyl, (C2 to Cg) cycloheteroalkyl, (C6 to Ci4) aryl, (C2 to Cg) heteroaryl, -(Ci to C3 alkylene)-(C3 to CB) cyclcoalkyl, -(Ci to C3 alkylene)- (C2 to Cg cycloheteroalkyl), -(Ci to C3 alkylene)- (C6 to C14. aryl) or -(Ci to C3 alkylene)- (C2 to C9 heteroaryl), each R16a or R16b is optionally further substituted by 1-3 R17; or, R16a and R16b, taken together with the nitrogen atom to which they are bound, form a (C2 to C9) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C2 to Cg) cycloheteroalkyl and (C2 to Cg) heteroaryl group is optionally further substituted by 1-3 R17; each R17 is independenly halogen, -OH1 cyano, -S(O)2-(Ci to C3 alkyl), (Ci to C3) alkyl, (Ci to C3) perfluoroalkyl (Ci to C3) alkoxy, -C(O)-(Ci-C6 alkyl), -C(O)-O(Ci to C6 alkyl), -C(O)-NH2, -C(O)-NH(Ci to C6 alkyl) Or - C(O)N(C1 to C6 alkyl)2; each p is independently O, 1, 2 or 3; provided when X is CR9, R9 is H, and each R11a, R11b and R11° is unsubstituted (Ci to C6) alkyl, R10 is not -C(O)-OR12; or a pharmaceutically accpetalbe salt thereof.
In one preferred aspect of the compound of formula (II) or salt thereof, X is CR9. More preferably, R9 is H, -OH or (Ci to C6) alkoxy. Even more preferably, R9 is -OH or (Ci to C6) alkoxy. EΞven more preferably, R9 is (Ci to C6) alkoxy. In another preferred aspect of the compound of formula (II) or salt thereof, X is N.
In another preferred aspect of the compound of formula (II) or salt thereof, R10 is
-C(O)NR13aR13b. More preferably, each R13a and R13b is independently H or (Ci to C6) alky wherein said (Ci to C6) alkyl is optionally further substituted by 1-3 halogen; even more preferably, R13a is H. Also more preferably, R13a and R13b, taken together with the nitrogen atom to which they are bound, form a (C-2 to C9) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C2 to Cg) cycloheteroalkyl or (C2 to Cg) heteroaryl group is optionally further substituted by 1-3 R14.
In another preferred aspect of the compound of formula (II) or salt thereof, R10 is -C(O)-OR12. More preferably, R12 is selected from (Ci to C6) alkyl, the said (Ci to C6) alkyl is optionally further substituted by 1-3 halogen.
In another preferred aspect of the compound of formula (II) or salt thereof, R10 is H.
In another preferred aspect of the compound of formula (II) or salt thereof, R10 is (Ci to C6) alkyl optionally further substituted by 1-3 halogen.
In another preferred aspect of the compound of formula (II) or salt thereof, R11a and R11 b is independently halogen, Ci to C6 alkyl or Ci to Cβ perfluoroalkyl, R11c is -O- (Ci to C6 alkylene)-R15, -0-(C2 to C6 alkenylene)-R15 or -0-(C2 to C6 alkynylene)-R15, R15 is (C2 to C9) heteroaryl, (C3 to Ce) cycloalkyl or (C2 to C9) heteroaryl, and R15 is optionally further substituted by 1-3 R17. More preferably, R11a and R11 b is independently halogen, R11c is -O-(Ci to C6 alkylene)-(C2 to C9 heteroaryl), wherein the (C2 to C9) heteroaryl is optionally further substituted by 1-3 groups selected from halogen and (Ci to C3) alkyl.
In a preferred embodiment of the compound of formula (II) or salt thereof,
X is CR9 or N;
R9 is H, -OH, or (C1-C3 alkoxy);
R10 is selected from (a) H, (Ci to C6) alkyl, and (Ci to C6) perfluoroalkyl,
(b) (Ci to C6) alkyl, further substituted by 1-3 groups selected from halogen and cyano,(c) (C3 to C8 cycloalkyl), (C2 to C9) cycloheteroalkyl, phenyl and (C2 to C9) heteroaryl, all optionally further substitued by 1-3 groups selected from (Ci to C6) alkyl, halogen, -OH, cyano, (Ci to C6) alkoxy, -S(O)2-(Ci to C3 alkyl), -NH2, -NH-(Ci to C3 alkyl) and -N(Ci to C3 alkyl)2, and
(d) -(CH2)p-C(O)-OR12, wherein R12 is H or (Ci to C6) alkyl optionally substituted by 1-3 halogen; and (e) -(CH2)p-C(O)NR13aR13b, wherein each R13a and R13b is selected from H, (Ci to C6 alkyl) optionally substituted by 1-3 halogen, (C3 to C8) cycloalkyl optionally substituted by 1-3 groups selected from halogen and (Ci to C3) alkyl, or R13a and R13b, taken together with the nitrogen atom to which they are bound, form a (C2 to C9) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C2 to C9) cycloheteroalkyl or (C2 to Cg) heteroaryl group is optionally further substituted by 1-3 groups selected from halogen, (Ci to C3) alkyl, (Ci-C3) perfluoroalkyl and -S(O)2-(Ci to C3 alkyl); each R11a and R11b is independently halogen, -OH1 (Ci to C6 alkyl), (C3 to Ce) cycloalkyl or (Ci to C6) alkoxy;
R11c is -0-(C1 to C6 alkylene)-R15, -0-(C2 to C6 alkenylene)-R15 or -0-(C2 to C6 alkynylene)-R15, R15 is (C2 to C9) heteroaryl, (C3 to C8) cycloalkyl or (C2 to C9) heteroaryl, and R15 is optionally further substituted by 1-3 R17.
In the previous preferred embodiment of the compound of formula (II) or salt thereof, more preferably, R10 is -C(O)NR13aR13b, wherein R13a is H, R13b is (Ci to C6 alkyl) optionally substituted by 1-3 halogen, or R13a and R13b, taken together with the nitrogen atom to which they are bound, form a (C2 to C9) cycloheteroalkyl group, the said (C2 to Cg) cycloheteroalkyl group is optionally further substituted by 1-3 groups selected from halogen, (Ci to C3) alkyl, (CrC3) perfluoroalkyl and -S(O)2-(Ci to C3 alkyl); each R11a and R11b is independently halogen, (Ci to C6 alkyl) or (Ci to C6) alkoxy;
R110 is -O-(Ci to C6 alkylene)-R15 or -0-(C2 to C6 alkenylene)-R15, R15 is (C2 to C9) heteroaryl, and R15 is optionally further substituted by 1-3 groups seleted from halogen and (Ci to C3) alkyl. It should also be understood that the present invention encompasses compounds of formula (II) as described above formed by any and all combinations of the preferred aspects, more preferred aspects, and even more preferred aspects of the compounds of formula (II), wherein any of the value of R9, R10, R11a, R11b, R11°, R12, R13a, R13b, R14, R15, R16a, R16b and R17 are further narrowed, so long as the aspects are not inconsistent with each other, and includes pharmaceutically acceptable salts and solvates thereof of any of said compounds. Also provided in the invention is a pharmaceutical composition, comprising a compound of formula (II) or a salt thereof. Also provided is a method of reducing abnormal cell growth in a mammal in need thereof, comprising the step of administering to said mammal a therapeutically effective amount of at least one compound of formula (II) or a salt thereof, more specifically, the abnormal cell growth is cancer. Also provided is the use of the compound of claim 1 , or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of cancer in a mammal.
The present invention further provides a pharmaceutical composition, comprising at least one compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier or diluent.
The present invention further provides a method of reducing abnormal cell growth in a mammal in need thereof, comprising the step of administering to said mammal a therapeutically effective amount of at least one compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof.
In one embodiment the abnormal cell growth is cancerous.
The present invention further provides a method of treating cancer in a mammal, comprising the step of administering to said mammal a therapeutically effective amount of at least one compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof.
The present invention further provides a method of inhibiting HSP-90 enzymatic activity, comprising contacting a HSP-90 enzyme with an HSP-90-inhibiting amount of at least one compound according to any one of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof. The present invention further provides the use of a compound according to any of the compounds described herein, or a pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for the treatment of abnormal cell growth in a mammal. In one embodiment, such abnormal cell growth is cancerous.
As used herein, the terms "comprising" and "including" are used in their open, non-limiting sense.
The terms "halo" and/or "halogen" refer to fluorine, chlorine, bromine or iodine. The term "(Ci to C6) alkyl" refers to a saturated aliphatic hydrocarbon radical including straight chain and branched chain groups of 1 to 6 carbon atoms. Examples of (Ci to C6) alkyl groups include methyl, ethyl, propyl, 2-propyl, n-butyl, /so-butyl, tert- butyl, pentyl, and the like. Similarly, terms such as "(C2 to C6) alkyl", "(C3 to C6) alkyl", and "(C1 to C4) alkyl" refer to the corresponding saturated aliphatic hydrocarbon radicals of 2 to 6 carbon atoms, 3 to 6 carbon atoms, and 1 to 4 carbon atoms, respectively.
The term "perfluoroalkyl" refers to an alkyl group in which all of its hydrogen atoms are replaced by fluorine atoms. The term "(C2 to Ce) alkenyl", as used herein, means an alkyl moiety comprising
2 to 8 carbons having at least one carbon-carbon double bond. The carbon-carbon double bond in such a group may be anywhere along the 2 to 8 carbon chain that will result in a stable compound. Such groups include both the E and Z isomers of said alkenyl moiety. Examples of such groups include, but are not limited to, ethenyl, propenyl, butenyl, allyl, and pentenyl. The term "allyl," as used herein, means a - CH2CH=CH2 group. The term, "C(R)=C(R)," as used herein, represents a carbon- carbon double bond in which each carbon is substituted by an R group.
As used herein, the term "(C2 to Cs) alkynyl" means an alkyl moiety comprising from 2 to 8 carbon atoms and having at least one carbon-carbon triple bond. The carbon-carbon triple bond in such a group may be anywhere along the 2 to 8 carbon chain that will result in a stable compound. Examples of such groups include, but are not limited to, ethyne, propyne, 1-butyne, 2-butyne, 1-pentyne, 2-pentyne, 1-hexyne, 2- hexyne, and 3-hexyne.
The term "(Ci to Cs) alkoxy", as used herein, means an O-alkyl group wherein said alkyl group contains from 1 to 8 carbon atoms and is straight, branched, or cyclic. Examples of such groups include, but are not limited to, methoxy, ethoxy, n-propyloxy, iso-propyloxy, n-butoxy, iso-butoxy, tert-butoxy, cyclopentyloxy, and cyclohexyloxy.
The term "(Ci to C8) heteroalkyl" refers to a straight- or branched-chain alkyl group having a total of from 2 to 12 atoms in the chain, including from 1 to 8 carbon atoms, and one or more atoms of which is a heteroatom selected from S, O, and N, with the proviso that said chain may not contain two adjacent O atoms or two adjacent S atoms. The S atoms in said chains may be optionally oxidized with one or two oxygen atoms, to afford sulfides and sulfones, respectively. Furthermore, the (Ci to Ce) heteroalkyl groups in the compounds of the present invention can contain an oxo group at any carbon or heteroatom that will result in a stable compound. Exemplary (Ci to C3) heteroalkyl groups include, but are not limited to, alcohols, alkyl ethers, primary, secondary, and tertiary alkyl amines, amides, ketones, esters, sulfides, and sulfones.
The term "{CQ to Cu) aryl", as used herein, means a group derived from an aromatic hydrocarbon containing from 6 to 14 carbon atoms. Examples of such groups include, but are not limited to, phenyl or naphthyl. The terms "Ph" and "phenyl," as used herein, mean a -CβH5 group. The term "benzyl," as used herein, means a -CH2C6H5 group.
"(C2 to Cg) heteroaryl", as used herein, means an aromatic heterocyclic group having a total of from 5 to 10 atoms in its ring, and containing from 2 to 9 carbon atoms and from one to four heteroatoms each independently selected from O, S and N, and with the proviso that the ring of said group does not contain two adjacent O atoms or two adjacent S atoms. The heterocyclic groups include benzo-fused ring systems. Examples of aromatic heterocyclic groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazaπyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. The C2 to Cg heteroaryl groups may be C-attached or N-attached where such is possible. For instance, a group derived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). Further, a group derived from imidazole may be imidazol-1-yl (N-attached) or imidazol-3-yl (O attached).
"(C-2 to Cg) cycloheteroalkyl", as used herein, means a non-aromatic, monocyclic, bicyclic, tricyclic, spirocyclic, or tetracyclic group having a total of from 4 to 13 atoms in its ring system, and containing from 2 to 9 carbon atoms and from 1 to 4 heteroatoms each independently selected from O, S and N, and with the proviso that the ring of said group does not contain two adjacent O atoms or two adjacent S atoms. Furthermore, such C2 to Cg cycloheteroalkyl groups may contain an oxo substituent at any available atom that will result in a stable compound. For example, such a group may contain an oxo atom at an available carbon or nitrogen atom. Such a group may contain more than one oxo substituent if chemically feasible. In addition, it is to be understood that when such a C2 to C9 cycloheteroalkyl group contains a sulfur atom, said sulfur atom may be oxidized with one or two oxygen atoms to afford either a sulfoxide or sulfone. An example of a 4 membered cycloheteroalkyl group is azetidinyl (derived from azetidine). An example of a 5 membered cycloheteroalkyl group is pyrrolidinyl. An example of a 6 membered cycloheteroalkyl group is piperidinyl. An example of a 9 membered cycloheteroalkyl group is indolinyl. An example of a 10 membered cycloheteroalkyl group is 4H- quinolizinyl. Further examples of such C2 to C9 cycloheteroalkyl groups include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1 ,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1 ,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl quinolizinyl, 3-oxopiperazinyl, 4-methylpiperazinyl, 4-ethylpiperazinyl, and 1-oxo- 2,8,diazaspiro[4.5]dec-8-yl.
The term "(C3 to C10) cycloalkyl group" means a saturated or unsaturated, monocyclic, fused, spirocyclic, or polycyclic ring structure having a total of from 3 to 8 carbon ring atoms. Examples of such groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, and adamantyl.
Similarly, terms such as "(C3 to C6) cycloalkyl" or "(C4 to C8) cycloalkyl" refer to the corresponding saturated or unsaturated, monocyclic, fused, spirocyclic, or polycyclic ring structure having a total of from 3 to 6 carbon ring atoms or from 4 to 8 carbon atoms, respectively.
The term "cyano" refers to a -C≡N group.
When "ene" is added after the "yl" at the end of any of the previously defined terms to form a new term, the new term refers to a diradical formed by removing one hydrogen atom from the original term of which the new term derived from. For example, an alkylene refers to a diradical group formed by removing one hydrogen atom from an alkyl group and that a "methylene" refers to a divalent radical -CH2- derived from removing one hydrogen atom from methyl. More examples of such diradicals include, but are not limited to: alkenylene, alkynylene, cycloalkylene, phenylene, heterocyclylene, heteroarylene and (nonaromatic unsaturated carbocyclylene), which are derived from alkenyl, alkynyl, cycloalkyl, phenyl, heterocyclyl, heteroary! and (nonaromatic unsaturated carbocyclyl), respectively. For example, "cyclopropylene"
Figure imgf000018_0001
For example, "CrC2 alkylene" refers to all of the following: -CH2-, -CH(CH3)- and -CH2-CH2-.
The term "substituted," means that the specified group or moiety bears one or more substituents. The term "unsubstituted," means that the specified group bears no substituents. The term "optionally substituted" means that the specified group is unsubstituted or substituted by one or more substituents. It is to be understood that in the compounds of the present invention when a group is said to be "unsubstituted," or is "substituted" with fewer groups than would fill the valencies of all the atoms in the compound, the remaining valencies on such a group are filled by hydrogen. For example, if a Ce aryl group, also called "phenyl" herein, is substituted with one additional substituent, one of ordinary skill in the art would understand that such a group has 4 open positions left on carbon atoms of the Ce aryl ring (6 initial positions, minus one to which the remainder of the compound of the present invention is bonded, minus an additional substituent, to leave 4). In such cases, the remaining 4 carbon atoms are each bound to one hydrogen atom to fill their valencies. Similarly, if a Cβ aryl group in the present compounds is said to be "disubstituted," one of ordinary skill in the art would understand it to mean that the Ce aryl has 3 carbon atoms remaining that are unsubstituted. Those three unsubstituted carbon atoms are each bound to one hydrogen atom to fill their valencies. The term "solvate", as used herein, means a pharmaceutically acceptable solvate form of a compound of the present invention that retains the biological effectiveness of such compound. Examples of solvates include, but are not limited to, compounds of the invention in combination with water, isopropanol, ethanol, methanol, dimethylsulfoxide (DMSO), ethyl acetate, acetic acid, ethanolamine, or mixtures thereof. It is specifically contemplated that in the present invention one solvent molecule can be associated with one molecule of the compounds of the present invention, such as a hydrate. Furthermore, it is specifically contemplated that in the present invention, more than one solvent molecule may be associated with one molecule of the compounds of the present invention, such as a dihydrate. Additionally, it is specifically contemplated that in the present invention less than one solvent molecule may be associated with one molecule of the compounds of the present invention, such as a hemihydrate. Furthermore, solvates of the present invention are contemplated as solvates of compounds of the present invention that retain the biological effectiveness of the non-hydrate form of the compounds.
The term "pharmaceutically acceptable salt," as used herein, means a salt of a compound of the present invention that retains the biological effectiveness of the free acids and bases of the specified derivative and that is not biologically or otherwise undesirable.
The term "pharmaceutically acceptable formulation," as used herein, means a combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a carrier, diluent, and/or excipient(s) that are compatible with a compound of the present invention, and is not deleterious to the recipient thereof. Pharmaceutical formulations can be prepared by procedures known to those of ordinary skill in the art. For example, the compounds of the present invention can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, and the like. Examples of excipients, diluents, and carriers that are suitable for such formulations include the following: fillers and extenders such as starch, sugars, mannitol, and silicic derivatives; binding agents such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl pyrrolidone; moisturizing agents such as glycerol; disintegrating agents such as povidone, sodium starch glycolate, sodium carboxymethylcellulose, agar, calcium carbonate, and sodium bicarbonate; agents for retarding dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as cetyl alcohol, glycerol monostearate; adsorptive carriers such as kaolin and bentonite; and lubricants such as talc, calcium and magnesium stearate and solid polyethylene glycols. Final pharmaceutical forms may be pills, tablets, powders, lozenges, saches, cachets, or sterile packaged powders, and the like, depending on the type of excipient used. Additionally, it is specifically contemplated that pharmaceutically acceptable formulations of the present invention can contain more than one active ingredient. For example, such formulations may contain more than one compound according to the present invention. Alternatively, such formulations may contain one or more compounds of the present invention and one or more additional agents that reduce abnormal cell growth.
The term "HSP-90-inhibiting amount" as used herein, refers to the amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, required to inhibit the enzymatic activity of HSP-90 in vivo, such as in a mammal, or in vitro. The amount of such compounds required to cause such inhibition can be determined without undue experimentation using methods described herein and those known to those of ordinary skill in the art.
The term "inhibiting HSP-90 enzyme activity," as used herein, means decreasing the activity or functioning of the HSP-90 enzyme either in vitro or in vivo, such as in a mammal, such as a human, by contacting the enzyme with a compound of the present invention.
The term "therapeutically effective amount," as used herein, means an amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, that, when administered to a mammal in need of such treatment, is sufficient to effect treatment, as defined herein. Thus, a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, is a quantity sufficient to modulate or inhibit the activity of the HSP-90 enzyme such that a disease condition that is mediated by activity of the HSP-90 enzyme is reduced or alleviated.
The terms "treat", "treating", and "treatment" refer to any treatment of an HSP-90 mediated disease or condition in a mammal, particularly a human, and include: (i) preventing the disease or condition from occurring in a subject which may be predisposed to the condition, such that the treatment constitutes prophylactic treatment for the pathologic condition; (ii) modulating or inhibiting the disease or condition, i.e., arresting its development; (iii) relieving the disease or condition, i.e., causing regression of the disease or condition; or (iv) relieving and/or alleviating the disease or condition or the symptoms resulting from the disease or condition, e.g., relieving an inflammatory response without addressing the underlying disease or condition. With regard to cancer, these terms simply mean that the life expectancy of an individual affected with a cancer will be increased or that one or more of the symptoms of the disease will be reduced.
The term "compound of the present invention" refers to any of the above- mentioned compounds, as well as those in the Examples that follow, and include those generically described or those described as species. The term also refers to pharmaceutically acceptable salts or solvates of these compounds.
"Abnormal cell growth", as used herein, unless otherwise indicated, refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition), including the abnormal growth of normal cells and the growth of abnormal cells. This includes, but is not limited to, the abnormal growth of: tumor cells (tumors) that proliferate by expressing a mutated tyrosine kinase or overexpression of a receptor tyrosine kinase; benign and malignant cells of other proliferative diseases in which aberrant tyrosine kinase activation occurs; any tumors that proliferate by receptor tyrosine kinases; any tumors that proliferate by aberrant serine/threonine kinase activation; benign and malignant cells of other proliferative diseases in which aberrant serine/threonine kinase activation occurs; tumors, both benign and malignant, expressing an activated Ras oncogene; tumor cells, both benign and malignant, in which the Ras protein is activated as a result of oncogenic mutation in another gene; benign and malignant cells of other proliferative diseases in which aberrant Ras activation occurs. Examples of such benign proliferative diseases are psoriasis, benign prostatic hypertrophy, human papilloma virus (HPV), and restinosis. "Abnormal cell growth" also refers to and includes the abnormal growth of cells, both benign and malignant, resulting from activity of the enzyme famesyl protein transferase.
The terms "abnormal cell growth" and "hyperproliferative disorder" are used interchangeably in this application.
The term "stereoisomers" refers to compounds that have identical chemical constitution, but differ with regard to the arrangement of their atoms or groups in space. In particular, the term "enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of one another. The terms "racemic" or "racemic mixture," as used herein, refer to a 1 :1 mixture of enantiomers of a particular compound. The term "diastereomers", on the other hand, refers to the relationship between a pair of stereoisomers that comprise two or more asymmetric centers and are not mirror images of one another.
Detailed Description The compounds of the present invention are useful for modulating or inhibiting
HSP-90 activity. Accordingly, these compounds are useful for the prevention and/or treatment of disease states associated with abnormal cell growth such as cancer, alone or in combination with other anti-cancer agents.
In accordance with a convention used in the art, the symbol ^ is used in structural formulas herein to depict the bond that is the point of attachment of the moiety or substituent to the core or backbone structure. In accordance with another convention, in some structural formulae herein the carbon atoms and their bound
hydrogen atoms are not explicitly depicted, e.g., '^- represents a methyl group,
SCH3 r reppnrrefisSefinnttss a ann e evtthhyvll g πrronui ipn,
Figure imgf000022_0001
represents a cyclopentyl group, etc. The compounds of the present invention may have asymmetric carbon atoms.
The carbon-carbon bonds of the compounds of the present invention may be depicted herein using a solid line ( ), a solid wedge ( m ), or a dotted wedge (--""111Hi).
The use of a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that all possible stereoisomers (e.g. specific enantiomers, racemic mixtures, etc.) at that carbon atom are included. The use of either a solid or dotted wedge to depict bonds to asymmetric carbon atoms is meant to indicate that only the stereoisomer shown is meant to be included. It is possible that compounds of the invention may contain more than one asymmetric carbon atom. In those compounds, the use of a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that all possible stereoisomers are meant to be included. For example, unless stated otherwise, it is intended that the compounds of the present invention can exist as enantiomers and diastereomers or as racemates and mixtures thereof. The use of a solid line to depict bonds to one or more asymmetric carbon atoms in a compound of the invention and the use of a solid or dotted wedge to depict bonds to other asymmetric carbon atoms in the same compound is meant to indicate that a mixture of diastereomers is present.
Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate using, for example, chiral high pressure liquid chromatography (HPLC). Alternatively, the racemate (or a racemic precursor) may be reacted with a suitable optically active compound, for example, an alcohol, or, in the case where the compound contains an acidic or basic moiety, an acid or base such as tartaric acid or 1- phenylethylamine. The resulting diastereomeric mixture may be separated by chromatography and/or fractional crystallization and one or both of the diastereoisomers converted to the corresponding pure enantiomer(s) by means well known to one skilled in the art. Chiral compounds of the invention (and chiral precursors thereof) may be obtained in enantiomerically-enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing from 0 to 50% isopropanol, typically from 2 to 20%, and from 0 to 5% of an alkylamine, typically 0.1% diethylamine. Concentration of the eluate affords the enriched mixture. Stereoisomeric conglomerates may be separated by conventional techniques known to those skilled in the art. See, e.g. "Stereochemistry of Organic Compounds" by E L Eliel (Wiley, New York, 1994), the disclosure of which is incorporated herein by reference in its entirety.
Where a compound of the invention contains an alkenyl or alkenylene group, geometric cisltrans (or ZIE) isomers are possible. Where the compound contains, for example, a keto or oxime group or an aromatic moiety, tautomeric isomerism ('tautomerism') can occur. Examples of tautomerism include keto and enol tautomers. A single compound may exhibit more than one type of isomerism. Included within the scope of the invention are all stereoisomers, geometric isomers and tautomeric forms of the inventive compounds, including compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof. Cis/trans isomers may be separated by conventional techniques well known to those skilled in the art, for example, chromatography and fractional crystallization.
Salts of the present invention can be prepared according to methods known to those of skill in the art. Examples of salts include, but are not limited to, acetate, acrylate, benzenesulfonate, benzoate (such as chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, and methoxybenzoate), bicarbonate, bisulfate, bisulfite, bitartrate, borate, bromide, butyne-1 ,4-dioate, calcium edetate, camsylate, carbonate, chloride, caproate, caprylate, clavulaπate, citrate, decanoate, dihydrochloride, dihydrogenphosphate, edetate, edislyate, estolate, esylate, ethylsuccinate, formate, fumarate, gluceptate, gluconate, glutamate, glycollate, glycollylarsanilate, heptanoate, hexyne-1 ,6-dioate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, γ-hydroxybutyrate, iodide, isobutyrate, isothionate, lactate, lactobionate, laurate, malate, maleate, malonate, mandelate, mesylate, metaphosphate, methane-sulfonate, methylsulfate, monohydrogenphosphate, mucate, napsylate, naphthalene-1 -sulfonate, naphthalene-2-sulfonate, nitrate, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, phenylacetates, phenylbutyrate, phenylpropionate, phthalate, phospate/diphosphate, polygalacturonate, propanesulfonate, propionate, propiolate, pyrophosphate, pyrosulfate, salicylate, stearate, subacetate, suberate, succinate, sulfate, sulfonate, sulfite, tannate, tartrate, teoclate, tosylate, triethiodode, and valerate salts.
The compounds of the present invention that are basic in nature are capable of forming a wide variety of different salts with various inorganic and organic acids. Although such salts must be pharmaceutically acceptable for administration to animals, it is often desirable in practice to initially isolate the compound of the present invention from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert the latter back to the free base compound by treatment with an alkaline reagent and subsequently convert the latter free base to a pharmaceutically acceptable acid addition salt. The acid addition salts of the base compounds of this invention can be prepared by treating the base compound with a substantially equivalent amount of the selected mineral or organic acid in an aqueous solvent medium or in a suitable organic solvent, such as methanol or ethanol. Upon evaporation of the solvent, the desired solid salt is obtained. The desired acid salt can also be precipitated from a solution of the free base in an organic solvent by adding an appropriate mineral or organic acid to the solution. Those compounds of the present invention that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include the alkali metal or alkaline-earth metal salts and particularly, the sodium and potassium salts. These salts are all prepared by conventional techniques. The chemical bases which are used as reagents to prepare the pharmaceutically acceptable base salts of this invention are those which form non-toxic base salts with the acidic compounds of the present invention. Such non-toxic base salts include those derived from such pharmacologically acceptable cations as sodium, potassium calcium and magnesium, etc. These salts can be prepared by treating the corresponding acidic compounds with an aqueous solution containing the desired pharmacologically acceptable cations, and then evaporating the resulting solution to dryness, preferably under reduced pressure. Alternatively, they may also be prepared by mixing lower alkanolic solutions of the acidic compounds and the desired alkali metal alkoxide together, and then evaporating the resulting solution to dryness in the same manner as before. In either case, stoichiometric quantities of reagents are preferably employed in order to ensure completeness of reaction and maximum yields of the desired final product.
If the inventive compound is a base, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.
If the inventive compound is an acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include organic salts derived from amino acids, such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium. In the case of agents that are solids, it is understood by those skilled in the art that the inventive compounds, agents and salts may exist in different crystal or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulas. The invention also includes isotopically-labeled compounds of the invention, wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as 2H and 3H, carbon, such as 11C, 13C and 14C, chlorine, such as 36CI, fluorine, such as 18F, iodine, such as 123I and 125I, nitrogen, such as 13N and 15N, oxygen, such as 15O, 17O and 18O, phosphorus, such as 32P, and sulfur, such as 35S. Certain isotopically-labeled compounds of the invention, for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, 3H, and carbon-14, UC, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium, 2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as 11C, 18F, 15O and 13N1 can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy.
Isotopically-labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed.
The compounds of the present invention may be formulated into pharmaceutical compositions as described below in any pharmaceutical form recognizable to the skilled artisan as being suitable. Pharmaceutical compositions of the invention comprise a therapeutically effective amount of at least one compound of the present invention and an inert, pharmaceutically acceptable carrier or diluent.
To treat or prevent diseases or conditions mediated by HSP-90, a pharmaceutical composition of the invention is administered in a suitable formulation prepared by combining a therapeutically effective amount (i.e., an HSP-90 modulating, regulating, or inhibiting amount effective to achieve therapeutic efficacy) of at least one compound of the present invention (as an active ingredient) with one or more pharmaceutically suitable carriers, which may be selected, for example, from diluents, excipients and auxiliaries that facilitate processing of the active compounds into the final pharmaceutical preparations.
The pharmaceutical carriers employed may be either solid or liquid. Exemplary solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly, the inventive compositions may include time-delay or time-release material known in the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate or the like. Further additives or excipients may be added to achieve the desired formulation properties. For example, a bioavailability enhancer, such as Labrasol, Gelucire or the like, or formulator, such as CMC (carboxy-methylcellulose), PG (propyleneglycol), or PEG (polyethyleneglycol), may be added. Gelucire®, a semisolid vehicle that protects active ingredients from light, moisture and oxidation, may be added, e.g., when preparing a capsule formulation.
If a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form, or formed into a troche or lozenge. The amount of solid carrier may vary, but generally will be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampoule or vial or non-aqueous liquid suspension. If a semi-solid carrier is used, the preparation may be in the form of hard and soft gelatin capsule formulations. The inventive compositions are prepared in unit-dosage form appropriate for the mode of administration, e.g., parenteral or oral administration.
To obtain a stable water-soluble dose form, a pharmaceutically acceptable salt of a compound of the present invention may be dissolved in an aqueous solution of an organic or inorganic acid, such as a 0.3 M solution of succinic acid or citric acid. If a soluble salt form is not available, the agent may be dissolved in a suitable co-solvent or combinations of co-solvents. Examples of suitable co-solvents include alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerin and the like in concentrations ranging from 0-60% of the total volume. In an exemplary embodiment, a compound of Formula I is dissolved in DMSO and diluted with water. The composition may also be in the form of a solution of a salt form of the active ingredient in an appropriate aqueous vehicle such as water or isotonic saline or dextrose solution.
Proper formulation is dependent upon the route of administration selected. For injection, the agents of the compounds of the present invention may be formulated into aqueous solutions, preferably in physiologically compatible buffers such as Hanks solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
For oral administration, the compounds can be formulated by combining the active compounds with pharmaceutically acceptable carriers known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated. Pharmaceutical preparations for oral use can be obtained using a solid excipient in admixture with the active ingredient (agent), optionally grinding the resulting mixture, and processing the mixture of granules after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include: fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; and cellulose preparations, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as crosslinked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, polyvinyl pyrrolidone, Carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active agents. Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active agents may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.
For administration intranasally or by inhalation, the compounds for use according to the present invention may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of gelatin for use in an inhaler or insufflator and the like may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch. The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit-dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active agents may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. In addition to the formulations described above, the compounds of the present invention may also be formulated as a depot preparation. Such long-acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion-exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt. A pharmaceutical carrier for hydrophobic compounds is a co- solvent system comprising benzyl alcohol, a non-polar surfactant, a water-miscible organic polymer, and an aqueous phase. The co-solvent system may be a VPD co- solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the non-polar surfactant polysorbate 80, and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD: 5W) contains VPD diluted 1 :1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. The proportions of a co-solvent system may be suitably varied without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity non-polar surfactants may be used instead of polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars or polysaccharides may be substituted for dextrose. Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide also may be employed, although usually at the cost of greater toxicity due to the toxic nature of DMSO. Additionally, the compounds may be delivered using a sustained- release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.
The pharmaceutical compositions also may comprise suitable solid- or gel-phase carriers or excipients. These carriers and excipients may provide marked improvement in the bioavailability of poorly soluble drugs. Examples of such carriers or excipients include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols. Furthermore, additives or excipients such as Gelucire®, Capryol®, Labrafil®, Labrasol®, Lauroglycol®, Plurol®, Peceol® Transcutol® and the like may be used.
Further, the pharmaceutical composition may be incorporated into a skin patch for delivery of the drug directly onto the skin.
It will be appreciated that the actual dosages of the agents of this invention will vary according to the particular agent being used, the particular composition formulated, the mode of administration, and the particular site, host, and disease being treated. Those skilled in the art using conventional dosage-determination tests in view of the experimental data for a given compound may ascertain optimal dosages for a given set of conditions. For oral administration, an exemplary daily dose generally employed will be from about 0.001 to about 1000 mg/kg of body weight, with courses of treatment repeated at appropriate intervals.
Furthermore, the pharmaceutically acceptable formulations of the present invention may contain a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, in an amount of about 0.1 mg to about 2000 mg, or from about 1 mg to about 1500 mg, or from about 5 mg to about 1000 mg, or from about 10 mg to about 750 mg, or from about 20 mg to about 500 mg, or from about 30 mg to about 500 mg, or from about 50 to about 500 mg, or from about 100 mg to about 500mg.
Additionally, the pharmaceutically acceptable formulations of the present invention may contain a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, in an amount from about 0.5 w/w% to about 95 w/w%, or from about 1 w/w% to about 95 w/w%, or from about 1 w/w% to about 75 w/w%, or from about 5 w/w% to about 75 w/w%, or from about 10 w/w% to about 75 w/w%, or from about 10 w/w% to about 50 w/w%.
The compounds of the present invention, or pharmaceutically acceptable salts or solvates thereof, may be administered to a mammal suffering from abnormal cell growth, such as a human, either alone or as part of a pharmaceutically acceptable formulation, once a day, twice a day, three times a day, or four times a day, or even more frequently.
Those of ordinary skill in the art will understand that with respect to the compounds of the present invention, the particular pharmaceutical formulation, the dosage, and the number of doses given per day to a mammal requiring such treatment, are all choices within the knowledge of one of ordinary skill in the art and can be determined without undue experimentation.
This invention also relates to a method for the treatment of abnormal cell growth in a mammal, including a human, comprising administering to said mammal an amount of any of the compound described herein, or a pharmaceutically acceptable salt or solvate thereof, that is effective in treating abnormal cell growth.
In one embodiment of this method, the abnormal cell growth is cancer, including, but not limited to, mesothelioma, hepatobilliary (hepatic and billiary duct), a primary or secondary CNS tumor, a primary or secondary brain tumor, lung cancer (NSCLC and SCLC), bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, gastrointestinal (gastric, colorectal, and duodenal), breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, testicular cancer, chronic or acute leukemia, chronic myeloid leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), primary CNS lymphoma, non hodgkins's lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocortical cancer, gall bladder cancer, multiple myeloma, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one or more of the foregoing cancers.
In one embodiment of the present invention the cancer is selected from lung cancer (NSCLC and SCLC), cancer of the head or neck, ovarian cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, breast cancer, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), primary CNS lymphoma, non hodgkins's lymphoma, spinal axis tumors, or a combination of one or more of the foregoing cancers.
In another embodiment of said method, said abnormal cell growth is a benign proliferative disease, including, but not limited to, psoriasis, benign prostatic hypertrophy or restinosis.
This invention also relates to a method for the treatment of abnormal cell growth in a mammal which comprises administering to said mammal an amount of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, that is effective in treating abnormal cell growth in combination with an anti-tumor agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxics, anti-hormones, and anti-androgens. In one embodiment of the present invention the anti-tumor agent used in conjunction with a compound of the present invention and pharmaceutical compositions described herein is an anti-angiogenesis agent, kinase inhibitor, pan kinase inhibitor or growth factor inhibitor. Preferred pan kinase inhibitors include Sutent™ (sunitinib), described in U.S. Patent No. 6,573,293. Anti-angiogenesis agents, include but are not limited to the following agents, such as EGF inhibitors, EGFR inhibitors, VEGF inhibitors, VEGFR inhibitors, TIE2 inhibitors, IGF1R inhibitors, COX-II (cyclooxygenase II) inhibitors, MMP-2 (matrix-metalloprotienase 2) inhibitors, and MMP-9 (matrix- metalloprotienase 9) inhibitors.
Preferred VEGF inhibitors, include for example, Avastin (bevacizumab), an anti- VEGF monoclonal antibody of Genentech, Inc. of South San Francisco, California. Additional VEGF inhibitors include CP-547,632 (Pfizer Inc., NY, USA), AG13736 (Pfizer Inc.), ZD-6474 (AstraZeneca), AEE788 (Novartis), AZD-2171 , VEGF Trap (Regeneron/Aventis), Vatalanib (also known as PTK-787, ZK-222584: Novartis & Schering AG), Macugen (pegaptanib octasodium, NX-1838, EYE-001 , Pfizer Inα/Gilead/Eyetech), IM862 (Cytran Inc. of Kirkland, Washington, USA); and aπgiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colorado) and Chiron (Emeryville, California) and combinations thereof.
VEGF inhibitors useful in the practice of the present invention are described in U.S. Patent No. 6,534,524 and 6,235,764, both of which are incorporated in their entirety for all purposes. Additional VEGF inhibitors are described in, for example in WO 99/24440, in WO 95/21613, WO 99/61422, U.S. Patent 5,834,504, WO 98/50356, U.S. Patent 5,883,113 U.S. Patent 5,886,020, U.S. Patent 5,792,783, U.S. Patent 6,653,308, WO 99/10349, WO 97/32856, WO 97/22596, WO 98/54093, WO 98/02438, WO 99/16755, and WO 98/02437, all of which are herein incorporated by reference in their entirety.
Other anti-angiogenic compounds include acitretin, fenretinide, thalidomide, zoledronic acid, angiostatin, aplidine, cilengtide, combretastatin A-4, endostatin, halofuginone, rebimastat, removab, Revlimid, squalamine, ukrain, Vitaxin and combinations thereof.
Other antiproliferative agents that may be used in combination with the compounds of the present invention include inhibitors of the enzyme farnesyl protein transferase and inhibitors of the receptor tyrosine kinase PDGFr, including the compounds disclosed and claimed in the following: U.S. Patent 6,080,769; U.S. Patent
6,194,438; U.S. Patent 6,258,824; U.S. Patent 6,586447; U.S. Patent 6,071,935; U.S.
Patent 6,495,564; and U.S. Patent 6,150,377; U.S. Patent 6,596,735; U.S. Patent
6,479,513; WO 01/40217; U.S. 2003-0166675. Each of the foregoing patents and patent applications is herein incorporated by reference in their entirety.
PDGRr inhibitors include but are not limited to those disclosed in international patent application publication numbers WO 01/40217 and WO 04/020431 , the contents of which are incorporated in their entirety for all purposes. Preferred PDGFr inhibitors include Pfizer's CP-673,451 and CP-868,596 and its salts. Preferred GARF inhibitors include Pfizer's AG-2037 (pelitrexol and its salts).
GARF inhibitors useful in the practice of the present invention are disclosed in U.S. Patent 5,608,082 which is incorporated in its entirety for all purposes. Examples of useful COX-II inhibitors which can be used in conjunction with a compound of Formula (I) and pharmaceutical compositions disclosed herein include CELEBREX™ (celecoxib), parecoxib, deracoxib, ABT-963, MK-663 (etoricoxib), COX- 189 (Lumiracoxib), BMS 347070, RS 57067, NS-398, Bextra (valdecoxib), paracoxib, Vioxx (rofecoxib), SD-8381, 4-Methyl-2-(3,4-dimethylphenyl)-1-(4-sulfamoyl-phenyl)-1H- pyrrole, 2-(4-Ethoxyphenyl)-4-methyl-1-(4-sulfamoylphenyl)-1H-pyrrole, T-614, JTE-522, S-2474, SVT-2016, CT-3, SC-58125 and Arcoxia (etoricoxib). Additonally, COX-II inhibitors are disclosed in U.S. patent applications US 2005-0148627 and US 2005- 0148777, the contents of which are incorporated in their entirety for all purposes. In a particular embodiment the anti-tumor agent is celecoxib (U.S. Patent
5,466,823), valdecoxib (U.S. Patent 5,633,272), parecoxib (U.S. Patent 5,932,598), deracoxib (U.S. Patent 5,521 ,207), SD-8381 (U.S. Patent 6,034,256, Example 175), ABT-963 (WO 02/24719), rofecoxib (CAS No. 162011-90-7), MK-663 (or etoricoxib) as disclosed in WO 98/03484, COX-189 (Lumiracoxib) as disclosed in WO 99/11605, BMS- 347070 (U.S. Patent 6,180,651), NS-398 (CAS 123653-11-2), RS 57067 (CAS 17932- 91-3), 4-Methyl-2-(3,4-dimethylphenyl)-1-(4-sulfamoyl-phenyl)-1 H-pyrrole, 2-(4- Ethoxyphenyl)-4-methyl-1-(4-sulfamoylphenyl)-1 H-pyrrole, or meloxicam.
Other useful inhibitors as anti-tumor agents used in combination with a compound of the present invention and pharmaceutical compositions disclosed herein include aspirin, and non-steroidal anti-inflammatory drugs (NSAIDs) which inhibit the enzyme that makes prostaglandins (cyclooxygenase I and II), resulting in lower levels of prostaglandins, include but are not limited to the following, Salsalate (Amigesic), Diflunisal (Dolobid), lbuprofen (Motrin), Ketoprofen (Orudis), Nabumetone (Relafen), Piroxicam (Feldene), Naproxen (Aleve, Naprosyn), Diclofenac (Voltaren), lndomethacin (Indocin), Sulindac (Clinoril), Tolmetin (Tolectin), Etodolac (Lodine), Ketorolac (Toradol), Oxaprozin (Daypro) and combinations thereof.
Preferred COX-I inhibitors include ibυprofen (Motrin), nυprin, naproxen (Aleve), indomethacin (Indocin), nabumetone (Relafen) and combinations thereof.
Targeted agents used in combination with a compound of the present invention and pharmaceutical compositions disclosed herein include EGFr inhibitors such as lressa (gefitinib, AstraZeneca), Tarceva (erlotinib or OSI-774, OSI Pharmaceuticals Inc.), Erbitux (cetuximab, lmclone Pharmaceuticals, Inc.), EMD-7200 (Merck AG), ABX-EGF (Amgen Inc. and Abgenix Inc.), HR3 (Cuban Government), IgA antibodies (University of Erlangen-Nuremberg), TP-38 (IVAX), EGFR fusion protein, EGF-vaccine, anti-EGFr immunoliposomes (Hermes Biosciences Inc.) and combinations thereof. Preferred EGFr inhibitors include Iressa, Erbitux, Tarceva and combinations thereof. Other anti-tumor agents include those selected from pan erb receptor inhibitors or
ErbB2 receptor inhibitors, such as CP-724,714 (Pfizer, Inc.), CM 033 (canertinib, Pfizer, Inc.), Herceptin (trastuzumab, Genentech Inc.), Omitarg (2C4, pertuzumab, Genentech Inc.), TAK-165 (Takeda), GW-572016 (lonafamib, GlaxoSmithKline), GW-282974 (GlaxoSmithKline), EKB-569 (Wyeth), PKI-166 (Novartis), dHER2 (HER2 Vaccine, Corixa and GlaxoSmithKline), APC8024 (HER2 Vaccine, Dendreon), anti-HER2/neu bispecific antibody (Decof Cancer Center), B7.her2.lgG3 (Agensys), AS HER2 (Research Institute for Rad Biology & Medicine), trifunctional bispecific antibodies (University of Munich) and mAB AR-209 (Aronex Pharmaceuticals Inc) and mAB 2B-1 (Chiron) and combinations thereof. Preferred erb selective anti-tumor agents include Herceptin, TAK-165, CP-
724,714, ABX-EGF, HER3 and combinations thereof. Preferred pan erbb receptor inhibitors include GW572016, CI-1033, EKB-569, and Omitarg and combinations thereof.
Additional erbB2 inhibitors include those disclosed in WO 98/02434, WO 99/35146, WO 99/35132, WO 98/02437, WO 97/13760, WO 95/19970, U.S. Patent 5,587,458, and U.S. Patent 5,877,305, each of which is herein incorporated by reference in its entirety. ErbB2 receptor inhibitors useful in the present invention are also disclosed in U.S. Patents 6,465,449, and 6,284,764, and in WO 01/98277 each of which are herein incorporated by reference in their entirety. Additionally, other anti-tumor agents may be selected from the following agents,
BAY-43-9006 (Onyx Pharmaceuticals Inc.), Genasense (augmerosen, Genta), Panitumumab (Abgenix/Amgen), Zevalin (Schering), Bexxar (Corixa/GlaxoSmithKline), Abarelix, Alimta, EPO 906 (Novartis), discodermolide (XAA-296), ABT-510 (Abbott), Neovastat (Aeterna), enzastaurin (EIi Lilly), Combrestatin A4P (Oxigene), 2D-6126 (AstraZeneca), flavopiridol (Aventis), CYC-202 (Cyclacel), AVE-8062 (Aventis), DMXAA (Roche/Antisoma), Thymitaq (Eximias), Temodar (temozolomide, Schering Plough) and Revilimd (Celegene) and combinations thereof. Other anti-tumor agents may be selected from the following agents, CyPat
(cyproterone acetate), Histerelin (histrelin acetate), Plenaixis (abarelix depot), Atrasentan
(ABT-627), Satraplatin (JM-216), thalomid (Thalidomide), Theratope, Temilifene (DPPE),
ABI-007 (paclitaxel), Evista (raloxifene), Atamestane (Biomed-777), Xyotax (polyglutamate paclitaxel), Targetin (bexarotine) and combinations thereof.
Additionally, other anti-tumor agents may be selected from the following agents, Trizaone (tirapazamine), Aposyn (exisulind), Nevastat (AE-941), Ceplene (histamine dihydrochloride), Orathecin (rubitecan), Virulizin, Gastrimmune (G17DT), DX-8951f (exatecan mesylate), Onconase (ranpirnase), BEC2 (mitumoab), Xcytrin (motexafin gadolinium) and combinations thereof.
Further anti-tumor agents may be selected from the following agents, CeaVac (CEA), NeuTrexin (trimetresate glucuronate) and combinations thereof. Additional antitumor agents may be selected from the following agents, OvaRex (oregovomab), Osidem (IDM-1), and combinations thereof. Additional anti-tumor agents may be selected from the following agents, Advexin (ING 201), Tirazone (tirapazamine), and combinations thereof. Additional anti-tumor agents may be selected from the following agents, RSR13 (efaproxiral), Cotara (131! chTNT 1/b), NBI-3001 (IL-4) and combinations thereof. Additional anti-tumor agents may be selected from the following agents, Canvaxin, GMK vaccine, PEG lnteron A, Taxoprexin (DHA/paciltaxel), and combinations thereof. Other anti-tumor agents include Pfizer's MEK1/2 inhibitor PD325901, Array
Biopharm's MEK inhibitor ARRY-142886, Bristol Myers' CDK2 inhibitor BMS-387,032, Pfizer's CDK inhibitor PD0332991 and AstraZeneca's AXD-5438, and combinations thereof.
Additionally, mTOR inhibitors may also be utilized such as CCI-779 (Wyeth) and rapamycin derivatives RAD001 (Novartis) and AP-23573 (Ariad), HDAC inhibitors, SAHA (Merck Inc./Aton Pharmaceuticals) and combinations thereof. Additional anti-tumor agents include aurora 2 inhibitor VX-680 (Vertex), and Chk1/2 inhibitor XL844 (Exilixis).
The following cytotoxic agents, e.g., one or more selected from the group consisting of epirubicin (Ellence), docetaxel (Taxotere), paclitaxel, Zinecard (dexrazoxane), rituximab (Rituxan) imatinib mesylate (Gleevec), and combinations thereof, may be used in combination with a compound of the present invention and pharmaceutical compositions disclosed herein. The invention also contemplates the use of the compounds of the present invention together with hormonal therapy, including but not limited to, exemestane (Aromasin, Pfizer Inc.), leuprorelin (Lupron or Leuplin, TAP/Abbott/Takeda), anastrozole (Arimidex, Astrazeneca), gosrelin (Zoladex, AstraZeneca), doxercalciferol, fadrozole, formestane, tamoxifen citrate (tamoxifen, Nolvadex, AstraZeneca), Casodex (AstraZeneca), Abarelix (Praecis), Trelstar, and combinations thereof.
The invention also relates to the use of the compounds of the present invention together with hormonal therapy agents such as anti-estrogens including, but not limited to fulvestrant, toremifene, raloxifene, lasofoxifene, letrozole (Femara, Novartis), anti- androgens such as bicalutamide, flutamide, mifepristone, nilutamide, Casodex™ (41- cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3'-(trifluoromethyl) propionanilide, bicalutamide) and combinations thereof.
Further, the invention provides a compound of the present invention alone or in combination with one or more supportive care products, e.g., a product selected from the group consisting of Filgrastim (Neupogen), ondansetron (Zofran), Fragmin, Procrit, Aloxi, Emend, or combinations thereof.
Particularly preferred cytotoxic agents include Camptosar, Erbitux, Iressa, Gleevec, Taxotere and combinations thereof.
The following topoisomerase I inhibitors may be utilized as anti-tumor agents: camptothecin; irinotecan HCI (Camptosar); edotecarin; orathecin (Supergen); exatecan (Daiichi); BN-80915 (Roche); and combinations thereof. Particularly preferred toposimerase Il inhibitors include epirubicin (Ellence).
Alkylating agents include, but are not limited to, nitrogen mustard N-oxide, cyclophosphamide, ifosfamide, melphalan, busulfan, mitobronitol, carboquone, thiotepa, ranimustine, nimustine, temozolomide, AMD-473, altretamine, AP-5280, apaziquone, brostallicin, bendamustine, carmustine, estramustine, fotemustine, glufosfamide, ifosfamide, KVV-2170, mafosfamide, and mitolactol; platinum-coordinated alkylating compounds include but are not limited to, cisplatin, Paraplatin (carboplatin), eptaplatin, lobapiatin, nedaplatin, Eloxatin (oxaliplatin, Sanofi) or satφlatin and combinations thereof. Particularly preferred alkylating agents include Eloxatin (oxaliplatin).
Antimetabolites include but are not limited to, methotrexate, 6-mercaptσpurine riboside, mercaptopurine, 5-fluorouracil (5-FU) alone or in combination with leucovorin, tegafur, UFT, doxifluridine, carmofur, cytarabine, cytarabine ocfosfate, enocitabine, S-1 , Alimta (premetrexed disodium, LY231514, MTA), Gemzar (gemcitabine, EIi Lilly), fludarabin, 5-azacitidine, capecitabine, cladribine, clofarabine, decitabine, eflornithine, ethynylcytidine, cytosine arabinoside, hydroxyurea, TS-1 , melphalan, nelarabine, nolatrexed, ocfosfate, disodium premetrexed, pentostatin, pelitrexol, raltitrexed, triapine, trimetrexate, vidarabine, vincristine, vinoreibine; or for example, one of the preferred anti-metabolites disclosed in European Patent Application No. 239362 such as N-(5-[N- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L- glutamic acid and combinations thereof. Antibiotics include intercalating antibiotics and include, but are not limited to: aclarubicin, actinomycin D, amrubicin, annamycin, adriamycin, bleomycin, daunorubicin, doxorubicin, elsamitrucin, epirubicin, galarubicin, idarubicin, mitomycin C, nemorubicin, neocarzinostatin, peplomycin, pirarubicin, rebeccamycin, stimalamer, streptozocin, valrubicin, zinostatin and combinations thereof. Plant derived anti-tumor substances include for example those selected from mitotic inhibitors, for example vinblastine, docetaxel (Taxotere), paclitaxel and combinations thereof.
Cytotoxic topoisomerase inhibiting agents include one or more agents selected from the group consisting of aclarubicn, amonafide, belotecan, camptothecin, 10- hydroxycamptothecin, 9-aminocamptothecin, diflomotecan, irinotecan HCI (Camptosar), edotecarin, epirubicin (Ellence), etoposide, exatecan, gimatecan, lurtotecan, mitoxantrone, pirarubicin, pixantrone, rubitecan, sobuzoxane, SN-38, tafluposide, topotecan, and combinations thereof.
Preferred cytotoxic topoisomerase inhibiting agents include one or more agents selected from the group consisting of camptothecin, 10-hydroxycamptothecin, 9- aminocamptothecin, irinotecan HCI (Camptosar), edotecarin, epirubicin (Ellence), etoposide, SN-38, topotecan, and combinations thereof.
Immunologicals include interferons and numerous other immune enhancing agents. Interferons include interferon alpha, interferon alpha~2a, interferon, alpha-2b, interferon beta, interferon gamma-1a, interferon gamma-1b (Actimmune), or interferon gamma-n1 and combinations thereof. Other agents include filgrastim, lentinan, sizofilan, TheraCys, ubenimex, WF-10, aldesleukin, alemtuzumab, BAM-002, dacarbazine, daclizumab, denileukin, gemtuzumab ozogamicin, ibritumomab, imiquimod, lenograstim, lentinan, melanoma vaccine (Corixa), molgramostim, OncoVAX-CL, sargramostim, tasonermin, tecleukin, thymalasin, tositumomab, Virulizin, Z-100, epratuzumab, mitυmomab, oregovomab, pemtumomab (Y-muHMFG1), Provenge (Dendreon) and combinations thereof.
Biological response modifiers are agents that modify defense mechanisms of living organisms or biological responses, such as survival, growth, or differentiation of tissue cells to direct them to have anti-tumor activity. Such agents include krestin, lentinan, sizofiran, picibanil, ubenimex and combinations thereof. Other anticancer agents that can be used in combination with a compound of the present invention include alitretinoin, ampligen, atrasentan bexarotene, bortezomib. Bosentan, calcitriol, exisulind, finasteride.fotemustine, ibandronic acid, miltefosine, mitoxantrone, l-asparaginase, procarbazine, dacarbazine, hydroxycarbamide, pegaspargase, pentostatin, tazarotne, Telcyta (TLK-286, Teiik Inc.), Velcade (bortemazib, Millenium), tretinoin, and combinations thereof.
Platinum-coordinated compounds include but are not limited to, cisplatin, carboplatin, nedaplatin, oxaliplatin, and combinations thereof.
Camptothecin derivatives include but are not limited to camptothecin, 10- hydroxycamptothecin, 9-aminocamptothecin, irinotecan, SN-38, edotecarin, topotecan and combinations thereof.
Other antitumor agents include mitoxantrone, l-asparaginase, procarbazine, dacarbazine, hydroxycarbamide, pentostatin, tretinoin and combinations thereof.
Anti-tumor agents capable of enhancing antitumor immune responses, such as CTLA4 (cytotoxic lymphocyte antigen 4) antibodies, and other agents capable of blocking CTLA4 may also be utilized, such as MDX-010 (Medarex) and CTLA4 compounds disclosed in U.S. Patent 6,682,736; and antiproliferative agents such as other famesyl protein transferase inhibitors, for example the farnesyl protein transferase inhibitors. Additionally, specific CTLA4 antibodies that can be used in combination with compounds of the present invention include those disclosed in U.S. Patents 6,682,736 and 6,682,736 both of which are herein incorporated by reference in their entirety. Specific IGF1R antibodies that can be used in the combination methods of the present invention include those disclosed in WO 02/053596, which is herein incorporated by reference in its entirety.
Specific CD40 antibodies that can be used in the present invention include those disclosed in WO 03/040170 which is herein incorporated by reference in its entirety. Gene therapy agents may also be employed as anti-tumor agents such as TNFerade (GeneVec), which express TNFalpha in response to radiotherapy.
In one embodiment of the present invention statins may be used in combination with a compound of the present invention and pharmaceutical compositions thereof. Statins (HMG-CoA reducatase inhibitors) may be selected from the group consisting of Atorvastatin (Lipitor™, Pfizer Inc.), Pravastatin (Pravachol™, Bristol-Myers Squibb), Lovastatin (Mevacor™, Merck Inc.), Simvastatin (Zocor™, Merck Inc.), Fluvastatin (Lescol™, Novartis), Cerivastatin (Baycol™, Bayer), Rosuvastatin (Crestor™, AstraZeneca), Lovostatin and Niacin (Advicor™, Kos Pharmaceuticals), derivatives and combinations thereof.
In a preferred embodiment the statin is selected from the group consisting of Atovorstatin and Lovastatin, derivatives and combinations thereof. Other agents useful as anti-tumor agents include Caduet. Methods of Preparation Compounds of the present invention may be prepared using the general procedures described below, employing the techniques available in the art using starting materials that are readily available. The preparation of certain embodiments of the present invention is described in detail in the following examples, but those of ordinary skill in the art will recognize that the preparations described may be readily adapted to prepare other embodiments of the present invention. For example, the synthesis of non-exemplified compounds according to the invention may be performed by modifications apparent to those skilled in the art, e.g., by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by making routine modifications of reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be recognized as having adaptability for preparing other compounds of the invention. In the following examples and preparations, "DMF" means di-methyl formamide,"DMAP" means di-methyl amino pyridine, "TBME" means tert-butyl methyl ether, "Me" means methyl, "TEA" means tri-ethyl amine, "i-PrOH" means isopropyl alcohol, "HATU" means O-(7-azabenzotriazol-1-yl)-N,N,N',N)-tetramethyluronium phosphorus pentafluoride, "DMSO" means di-methyl sulfoxide, "EtOAc" means ethyl acetate, "Et" means ethyl, "Ph" means phenyl, "Boc" means t-butyloxycarbonyl, "DCM" means di-chloro methane, "DME" means di-methyl ether, "MeOH" means methanol, "THF" means tetrahydrofuran, "DIEA" means diisopropyl ethylamine, "HOAc" means acetic acid, "NaOAc" means sodium acetate, "DMA" means dimethyl amide, "MTBE" means methyl tert-butyl ether, "TLC" means thin layer chromatography, and "TFA" means tri-fluoro acetic acid. General Procedure GA
Figure imgf000042_0001
R1, R2, R3 = halogen, optionally substituted alkyl, optionally substituted alkoxy, amino, carboxyamidθ, cyano, aryl or heteroaryl.
Synthesis of compound 2 (2-aminothienor2,3-dipyrimidin-4-ol)
To a solution of cyanamide (1.26 g, 30.0 mmol) in dioxane (10.0 ml_) was added 4N HCI in dioxane (10.0 mL, 40.0 mmol) dropwise. The resulting chloroformamidine (compound 1) precipitated and was stirred for 1 hour, then the solvent was removed under vacuum. The chloroformamidine (compound 1) was partially dissolved in diglyme (20.0 mL) and methyl 2-aminothiophene-3-carboxylate (1.89 g, 12.0 mmol) was added in one portion. This suspension was heated to reflux (2000C oil bath) until all solids dissolved as a pink-brown color solution (-10 minutes). The reaction mixture was then cooled to room temperature, and the precipitate was collected by filtration. The crude material was redisolved in hot (7O0C) 2N NaOH (50 mL), cooled to room temperature, neutralized with concentrated HCI to pH 2.3 and the precipitate collected by filtration to afford 941 mg of 2-aminothieno[2,3-d]pyrimidin-4-o! (compound 2) as a sand color solid (47% yield). 1H NMR (400 MHz, DMSOd6) δ ppm 6.53 (s, 2 H) 6.96 (d, J=5.81 Hz, 1 H) 7.08 (d, J=5.81 Hz, 1 H) 10.90 (s, 1 H).
Synthesis of compound 3 (N-(4-hvdroxythienor2,3-d1pyrimidin-2-yl)-2,2- dimethylpropanamide) A reaction mixture of 2-aminothieno[2,3-d]pyrimidin-4-ol (compound 2, 941 mg, 5.63 mmol), trimethylacetic anhydride/Piv2O (2.3 mL, 1 1.3 mmol), triethylamine (1.6 mL, 11.3 mmol) and DMAP (138 mg, 1.13 mmol) in 4mL of DMF was stirred at 8O0C for 16 hours. The reaction mixture was partitioned between EtOAc (500 mL) and saturated NaHCO3 (100 mL) and brine (100 mL). The combined organic layers were dried (Na2SO4) and then concentrated by vacuum. The residue was purified by Biotage system (4OM, EtOAc:hexane /0% to 30%) to collect the desired fractions of 861 mg of N-(4- hydroxythieno[2,3-d]pyrimidin-2-yl)-2,2-dimethylpropanamide (compound 3) as a white solid (61% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.32 (s, 9 H) 7.07 (d, J=5.81 Hz, 1 H) 7.41 (d, J=5.81 Hz, 1 H) 8.08 (s, 1 H). Synthesis of compound 4 (N-(4-chlorothienof2,3-dlpyrimidin-2-vi)-2,2- dimethylpropanamide)
A reaction mixture of N-(4-hydroxythieno[2,3-d]pyrimidin-2-yl)-2,2-dimethylpropanamide (compound 3, 861 mg, 1.48 mmol) with POCI3 (7.0 mL, 75.4mmol) was vigorously stirred at 1150C for 30 minutes while the reaction mixture became a brown homogenous color and then continued heating for an additional 30 minutes. The reaction mixture was quickly cooled to O0C in an ice/water bath, then basified with 2N NaOH to pH 7.2. The resulting white suspension was then filtered off and washed well with EtOAc (600 mL). The filtrate was extracted with brine (100 mL). The white solid was then filtered again and washed with H2O (100 mL) and CH2CI2 (300 mL). The organic layer was separated and then extracted with brine (100 mL). The combined organic layers were dried (Na2SO4) and then concentrated by vacuum. The residue was purified by Biotage system (40M1 EtOAc:hexane/0% to 30%) to collect desired #2 fractions to afford 754 mg of N-(4-chlorothieno[2,3-d]pyrimidin-2-yl)-2,2-dimethylpropanamide (compound 4) as a white solid (82% yield). 1H NMR (400 MHz, DMSOd6) δ ppm 1.23 (s, 9 H) 7.46 (d, J=6.06 Hz, 1 H) 7.89 (d, J=6.06 Hz, 1 H) 10.49 (s, 1 H). Synthesis of Compounds 5 and 6:
Compounds 5 and 6 can be prepared as described in Example 1. General Procedure GB
General Procedure GB1:
Figure imgf000044_0001
Synthesis of compound 1 was accomplished by following the procedure reported in J. Org. Chem. 60, pp. 7947-7952 (1995). Synthesis of compound 2 was accomplished by following the procedure reported in J. Med. Chem. 46, pp. 3060-3071 (2003). General Procedure GB2:
Figure imgf000044_0002
R1, R2, R3 = halogen, optionally substituted alkyl, optionally substituted alkoxy, amino, carboxyamide, cyano, aryl, or heteroaryl.
General Procedure GB2 can be carried out as described herein in Examples 2-6. General Procedure GB3:
Figure imgf000045_0001
Compound 3 from GB2 4 5
R= H, alkoxyl, amino, alkylcyano, fluoroalkyl, optionally substituted (C1-C^aHCyI1 fluoroalkyl, optionally substituted aryl or optionally substituted heteroaryl
General Procedure GB3 can be carried out as described herein in Examples 7 and 8. General Procedure GC:
Figure imgf000045_0002
R1, R2, R3 = halogen, optionally substitued alkyl, optionally substituted alkoxy, amino, carboxyamide, cyano, aryl or heteroaryl.
Synthesis of compound 1 (δ-amino-Σ-methyl-thiazole^-carboxylic acid ethyl ester): Acetamidocyanoacetate was dissolved/suspended in toluene (1.2 L), followed by the addition of Lawesson's reagent. The light yellow suspension was heated at 7O0C for 16 hours. The reaction mixture was cooled to room temperature. The toluene layer was separated from the gummy material at the bottom and extracted with 1 N HCl (6x500 mL). The combined aqueous layers were basified upon the addition of 2 N NaOH 1.9 L. The water layer was extracted with ethyl acetate (2x1.5 L), dried (Na2SO4), filtered and concentrated in vacuo, yielding crude compound 1 (20 g, 10.7 mmol, purity >99 %).
The gummy material was dissolved in methanol (500 mL) and concentrated under reduced pressure. The sticky product was dissolved in TBME (1.0 L) and 1 N HCI (1.5 L). The aqueous layer was basified with 2 N NaOH (1.0 L). The product was extracted with ethyl acetate (1.0 L), dried (Na2SO4), filtered and concentrated in vacuo, yielding compound 1 (50.3 g, 27.0 mmol, purity >99 %). Overall yield 65 % (70.7 g, 38.0 mmol, purity >99 %).
Synthesis of compound 2 (5-amino-2-methylthiazolor5,4-o1pyrimidin-7-ol): 5-amino-2-methyl-thiazole-4-carboxylic acid ethyl ester (compound 1) (31.3 g. 0.168 mol) and cyanamide (17.5 g, 0.416 mol) were dissolved in dioxane (150 ml_, pre-dried using 4 A molecular sieves). In addition, drop wise 4 N HCI in dioxane (200 ml) was added over 15 minutes and the mixture was heated to reflux and vigorously stirred, for 5 days. The volatiles were evaporated to dryness and the crude product (55 g) was used without any further purification in the next step.
Synthesis of compound 3 (Λ/-(7-hydroxy-2-methylthiazolo['5,4-Qlpyrimidin-5- vDpivalamide):
The crude product compound 2 (55 g) was dissolved in DMF (300 mL). Triethyl amine
(105 mL, 0.755 mol) was added followed by the addition of trimethyl acetic anhydride (92 mL, 453 mmol). The mixture was heated to reflux for 1 hour yielding a clear deep brown solution. TLC showed conversion to target compound and consumption of starting material. The volatiles were evaporated under reduced pressure. The remaining solid was purified by column chromatography (7% MeOH in CH2CI2), yielding compound 3 as an off-white solid (9.0 g, 20.7 mmol) in 20 % yield over two steps. Synthesis of compound 4 (Λ/-(7-chloro-2-methylthiazolor5,4-c/|PVrimidin-5-yl)pivalamide): Λ/-(7-hydroxy-2-methylthiazolo[5,4-c(]pyrimidin-5-yl)pivalamide (compound 3) (5.5 g, 20.7 mmol) was added portion wise to POCI3 (60 mL). The mixture suspension was heated to reflux for 1.5 hours and the suspension changed into a dark brown solution. The mixture was cooled to ambient temperature and poured drop wise on 1 kg of crushed ice/water. The mixture was neutralized to pH = 7 by the batch wise addition of 30% NaOH aq. (~180 mL). The product was extracted with dichloromethane (2x500 mL), dried (Na2SO4) and concentrated in vacuo, yielding compound 4 (4.45 g,15.6 mmol) in 75% yield. Compounds 5 and 6 can be prepared as described herein in Examples 9, 10, and 11. General Procedure GD (for boronic acids/esters synthesis): Example GD-1 :
Figure imgf000047_0001
Synthesis of compound 1 :
Into a 2 L 3-neck flask was added BF3 OEt2 (70.6 mL, 79.7 g, 560 mmol) and the system was cooled to O0C. To this system was added a solution of 5-amino-2,4-dichlorophenol (50.0 g, 280 mmol) in 700 mL of THF over 45 minutes. Upon completion of addition, a solution of isoamyl nitrite (48.8 mL, 42.78 g, 365 mmol) in 150 mL of THF was added over 15 minutes. The reaction was stirred an additional 30 minutes at O0C during which time a yellow precipitate formed. Et2O was added to increase precipitation and the yellow solid was filtered, washed with Et2O, and air dried. The precipitate was then added to a solution of NaI (54.7 g, 365 mmol) in 1.2 L of acetone portionwise and the red solution was stirred at ambient temperature overnight. Solvent was removed by rotary evaporation leaving an orange semisolid. Water was added and the mixture was extracted 2x with EtOAc. The combined organics were washed with saturated NaHSθ3 and dried over Na2SO4. After filtration of the drying agent, solvent was removed in vacuo leaving an orange oil. The oil was chromatographed through a plug of SiO2 by eluting with CH2CI2 and solvent was removed in vacuo leaving a mixture of compound 1 and the reduced compound 1a as yellow solid. The solid is recrystallized from hexanes (3 crops) leaving compound 1 (43.4 g, 150 mmol; 54% yield) as a yellow solid. Synthesis of compound 2: In a 1 L flask was dissolved compound 1 in 400 mL of CH2CI2 and the solution was cooled to O0C. To this solution was added methoxymethyl chloride (13.7 mL, 14.53 g, 180 mmol) followed by diisopropylethyl amine (31.5 mL, 23.3 g, 180 mmol) and the reaction was allowed to warm to ambient temperature overnight. The reaction was washed with saturated NaHCO3 and dried over Na2SO4. After filtration of the drying agent, solvent was removed in vacuo leaving compound 2 (46.8 g, 141 mmol, 94%) as a yellow solid. Synthesis of compound A: In a 500 mL flask were mixed compound 2 (41.8 g, 125 mmol), bis(pinacolato)diboron (35.1 g, 138 mmol), potassium acetate (24.6 g, 250 mmol), bis(diphenylphosphino)ferrocene-dichloropalladium dichloromethane adduct (6.15 g, 7.5 mmol), and 375 mL of dry dioxane. The mixture was degassed for 1 hour, then heated to 9O0C. After 5 days, complete conversion of compound 2 was observed by gas chromatography and the reaction was cooled to ambient temperature. The mixture was filtered, the solid washed with Et2O1 and solvent was removed by rotary evaporation leaving a brown residue. The residue was purified by chromatography (hexanes to 5:1 hexanes:EtOAc). Solvent was removed in vacuo leaving compound A (2-(2,4-dichloro- 5-methoxymethoxy-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxab) (30.2 g, 91 mmol, 73% yield; >94% purity by GCMS) as a yellow solid. 1H NMR (400 MHz, chloroform-cQ δ ppm 1.37 (s, 12 H) 3.53 (s, 3 H) 5.26 (s, 2 H) 7.39 (s, 1 H) 7.42 (s, 1 H). Example GD-2:
Figure imgf000048_0001
Synthesis of compound B:
Compound B (2-(2-chloro-3-methoxymethoxy-4-methyl-phenyl)-4,4,5,5-tetramethyl-
[1,3,2]dioxaborolane) was synthesized using the same procedure as in Example GD-1 using commercially available 2 3-Amino-2,6-dichloro-phenol as the starting material. 1H
NMR (400 MHz, chloroform-d) δ ppm 1.37 (s, 12 H) 2.36 (s, 3 H) 3.89 (q, J=7.07 Hz, 3
H) 5.12 (s, 2 H) 7.07 (d, J=7.58 Hz, 1 H) 7.36 (d, J=7.58 Hz, 1 H)
Example GD-3:
Figure imgf000049_0001
Synthesis of compound 1 (1 -chloro-2-iodo-3,5-dimethoxybenzene): To a reaction mixture of (2-chloro-4,6-dimethoxyphenyl)amine (1880 mg, 10.0 mmol) in 40.0 mL of acetic acid was added 6N HCI aqueous solution (10.0 nriL, 60.0 mmol). The resulting mixture was cooled with (ice/water/salt) and then another 2N aqueous solution of sodium nitrite (828 mg, 12.0 mmol in 6.0 mL of water) was added slowly (keeping the temperature <5°C). On complete addition the resulting solution was stirred for -30 minutes. To the resulting mixture was added a solution of potassium iodide (3320 mg, 20.0 mmol) and iodine (761 mg, 3.0 mmol) in 35 mL of water dropwise. The resulting mixture was stirred at O0C to room temperature for ~90 minutes. To the reaction mixture was added 120 mL of water and then extracted with EtOAc (2x300 mL). The combined extracts were washed with 10% aqueous Na2S2O3 solution (2x100 mL) and brine (100 mL). The organic layers were dried (Na2SO4) then concentrated by vacuum. The residue was purified by Biotage system (40M, EtOAc: Hexane/0% to 30%) to collect the desired fractions to afford 1.45g of 1-chloro-2-iodo-3,5-dimethoxybenzene as a yellowish solid (49% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 3.79 (s, 3 H) 3.84 (S1 3 H) 6.30 (d, J=2.53 Hz, 1 H) 6.68 (d, J=2.53 Hz, 1 H). Synthesis of compound C: A reaction solution of 1-chloro-2-iodo-3,5-dimethoxybenzene (1.45 g, 4.86 mmol), Pinacolborane (1.4 mL, 9.72 mmol) and Et3N (2.0 mL, 14.6.0 mmol) in 25.0 mL of Dioxane was purged with N2 for 15 minutes, then Pd(OAc)2 (55 mg, 0.243 mmol) and PCy2(O-BiPh) (170 mg, 0.7 mmol) were added. The resulting mixture was stirred at 8O0C for 4 hours. The reaction mixture was filtered through Celite pad and washed well with EtOAc. The filtrate was extracted with water (100 mL) and brine (100 mL). The organic layer was dried (Na2SO4) and then concentrated by vacuum. The residue was purified by Biotage system (40 M, EtOAc:Hexane / 0-20%) to collect the desired #2 fraction to afford 1.104 g as a yellowish solid (77% yield). 1H NMR (400 MHz, chloroform-D) δ ppm 1.37 (s, 12 H) 3.73 (s, 3 H) 3.76 (s, 3 H) 6.25 (s, 1 H) 6.44 (s, 1 H). Example GD-4:
Figure imgf000050_0001
D
Synthesis of compound D:
Compound D (2-(2,4-dichloro-5-isopropoxyphenyl)-4,4,5,5-tetramethyl-1 ,3,2- dioxaborolane) was synthesized using the same procedure as in Example GD-3 using commercially available 2,4-dichloro-5-isopropoxy-phenylamine as the starting material.
1H NMR (400 MHz, chloroform-d) δ ppm 1.26 - 1.33 (m, 18 H) 4.47 - 4.55 (m, 1 H) 7.17
(s, 1 H) 7.29 (s, 1 H).
Example GD-5:
Figure imgf000050_0002
Synthesis of compound E:
To a 20 mL microwave tube was charged 2-bromo-1 ,5-dimethoxy-3-methylbenzene (1.13 g, 4.89 mmol), Pinacolborane (2.13ml, 14.7mmol), Et3N (2.73 mL, 19.6 mmol) and 15 mL of Dioxane and then purged with N2 for 15 minutes. Pd(ll)CI2(PPh3)2 (343 mg, 0.489 mmol) was then added. The resulting mixture was microwaved at 1500C for 1 hour. The reaction mixture was filtered through Celite pad and washed well with EtOAc. The filtrate was extracted with water (2x50 mL) and brine (50 mL). The organic layer was dried (Na2SO4) and then concentrated by vacuum. The residue was purified by Biotage system (4OM, EtOAc.Hexane / 0-20%) to collect desired #2 fraction to afford
605 mg as yellow oil (45% yield). 1H NMR (400 MHz, chloroform-d) δ ppm 1.35 (s, 12
H) 2.33 (s, 3 H) 3.73 (s, 3 H) 3.76 (s, 3 H) 6.20 (d, J=2.02 Hz, 1 H) 6.27 (d, J=2.02 Hz, 1
H).
Example GD-6
Figure imgf000051_0001
Synthesis of Compound 1 :
To a solution of 3,5-dichloro-phenol (70 g, 0.43 mol) in dry toluene (1 L) was added NaH (51.5 g, 1.29 mol) portionwise at 0 D under N2 atmosphere. After the addition, the resulting mixture was allowed to warm up to room temperature and stirred for 20 minutes. The suspension was then cooled back to 0 D, and iodine (253.81 g, 91.5 mol) was added slowly. Then the reaction mixture was stirred at room temperature overnight. TLC (petroleum ether/CH2Cl2 1 :1) indicated complete consumption of starting material. The reaction mixture was quenched with 1 N HCI (1 L) and diluted with ether (1 L). The separated organic layer was washed with brine (500 mL), dried over Na2SO4 and concentrated in vacuo to give crude compound 2, which was purified by column chromatography (silica gel, petroleum ether/CH2CI2 from 5:1 to 1:1) to yield pure compound 1 (85 g, yield: 68%) as a white solid. Synthesis of Compound 2:
A mixture of compound 1 (67 g, 0.23 mol), chloromethoxy-ethane (31.8 g, 0.29 mol) and Cs2Cθ3 (63.7 g, 0.2 mol) in DMF (600 mL) was stirred at room temperature for 2 hours. TLC (petroleum ether/EtOAc 2:1) indicated complete consumption of compound 1. The reaction mixture was washed with H2O (500 mL x 3) and brine (500 mL), dried over Na2SO4 and concentrated in vacuo to give crude compound 3, which was purified via column chromatography (silica gel, EtOAc/hexane 1 :50) to yield pure compound 2 (80 g, 100%) as a yellow solid. Svntehsis of Compound 3:
A solution of compound 2 (77 g, 0.22 mol), 4,4,5,5-tetramethyl-[1,3,2]dioxaborolane (57 g, 0.44 mol) and Et3N (92 mL, 0.66 mol) in dioxane (500 ml_) was purged with N2 for 30 minutes. Then Pd(OAc)2 (2.7 g, 0.011 mol) and biphenyl-2-yl-dicyclohexyl-phosphane (8.5 g, 0.022 mol) were added to the resulting mixture. After the addition, the reaction mixture was stirred at 80 D for 1.5 hours. TLC (petroleum ether/EtOAc 30:1 ) indicated complete consumption of compound 2. The resulting mixture was washed with saturated NH4CI (500 mL), H2O (500 mL) and brine (500 mL) in sequence, dried over Na2SO4 and concentrated in vacuo to give crude compound 3, which was purified by column chromatography (silica gel, petroleum ether/EtOAc from 5:1 to 1 :1) to yield pure compound 3 (35 g, yield: 45%) as a brown solid. Synthesis of Compound F: To a stirred solution of compound 3 (35 g, 0.1 mol) in dry CH2CI2 (200 mL) was added BBr3 (125 g, 0.5 mol) dropwise at 0 □ under N2 atmosphere. After stirring for 20 minutes, the reaction mixture was poured into ice water, basified to pH ~ 10 by 3 N NaOH (100 mL) and the organic layer was separated. The separated aqueous layer was adjusted to pH ~ 3 with 1 N HCI (500 mL) and extracted with EtOAc (500 mL x 3), the combined organic layers were washed with brine (1.0 L), dried over Na2SO4 and concentrated in vacuo to yield compound F (39.7 g, yield: 80%) as a white solid. 1H NMR (400 MHz, MeOD): δ 6.875-6.878 (d, 1 H)1 6.727-6.737 (d, 1 H).
General Procedure GE (for derivatization of the bromo atom)
Figure imgf000052_0001
R= alkoxyl, amino, alkylcyano, fluoroalkyl, optionally substituted (C,-C6)a!kyl, optionally substituted aryl or optionally substituted heteroaryl
R'= H, alkyl, cyano, amino, fluoroalkyl, carboxyamide, or carboxylic acid Q and T = C, N, or S General Procedure GE can be carried out as described in Example 13.
Examples
The examples below are intended to illustrate particular embodiments of the present invention and are not meant to limit the scope of the invention in any way. Examples 1 to 13 provide detailed synthetic steps for preparing several specific compounds of the present invention. Table 1 shows additional compounds that were prepared as Examples 14 to 15 according to the general reaction schemes as described herein. Examples 16 and 17 describe the biochemical and cellular assays used to assess the potency of the compounds shown in Examples 1 to 15. Table 3 shows the biochemical and celluar assay values for compounds prepared as Examples 1 to 15.
The compounds shown in the examples below may have asymmetric carbon atoms. The carbon-carbon bonds of the compounds of the present invention may be depicted herein using a solid line ( ), a solid wedge ( -" ), or a dotted wedge ( "" ). In the compounds shown in the examples below, the use of a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that the compound was prepared as a racemic mixture, or that the compound was prepared as a pure enantiomer but the absolute stereochemistry was not determined. The use of either a solid or dotted wedge to depict bonds to asymmetric carbon atoms is meant to indicate that the compound was prepared as a stereoisomer where the absolute stereochemistry was determined.
Various starting materials and other reagents were purchased from commercial suppliers, such as Aldrich Chemical Company, and used without further purification, unless indicated otherwise. 1H-NMR spectra were recorded on a Bruker instrument operating at 400 or 500
MHz. NMR spectra were obtained as DMSO-dβ or CDCI3 solutions (reported in ppm), using chloroform as the reference standard (7.25 ppm and 77.00 ppm) or DMSOd6 (2.50 ppm and 39.52 ppm). Other NMR solvents were used as needed. When peak multiplicities are reported, the following abbreviations are used: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublets, dt = doublet of triplets. Coupling constants, when given, are reported in Hertz. Example 1 : 4-(2-chloro-4,6-dimethoxyphenyl)thieno[2,3-d]pyrimidiπ-2-amine
Figure imgf000054_0001
The above compound was prepared as follows using General Procedure GA as described previously. Compound C in General Procedure GD-3 (166 mg, 0.56 mmol) was added to a solution of compound 4 in General Procedure GA (150 mg, 0.56 mmol) in 10 mL of dioxane. The mixture was purged with N2 several times. Pd(PPr^)4 (65 mg, 0.056 mmol) was added then Na2CO3 (2 M, 1 mL) was added to the mixture. The mixture was heated and stirred at 850C for 12 hours. The reaction was cooled to room temperature and H∑O (20 mL) was added to the reaction mixture. EtOAc (2x 50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil. The residue was purified by silica gel chromatography (gradient elution 30→40% EtOAc in hexanes) to give a white solid (175 mg, 77.5% yield) as the desired intermediate product (compound 5a: N-[4-(2- chloro-4,6-dimethoxy-phenyl)-thieno[2,3-d]pyrimidin-2-yl]-2,2-dimethyl-propionamide).
Figure imgf000054_0002
5a Sodium hydroxide (10 eq., 4 mmol, 2 mL, 2M in H2O) was added to a solution of compound 5a, (168 mg, 0.41 mmol) in EtOH (4 mL). The mixture was heated to 9O0C and stirred for 12 hours. The solvent was evaporated and the solution was acidified with acetic acid buffer. EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to obtain a yellow wax residue as the desired final product (76 mg, 57% yield). 1H NMR (400 MHz, DMSO-d6) δ ppm 3.68 (s, 3 H) 3.85 (s, 3 H) 6.66 (d, J=5.81 Hz, 1 H) 6.70 (d, J=2.02 Hz, 1 H) 6.77 (d, J=2.02 Hz, 1 H) 6.83 (s, 2 H) 7.18 (d, J=6.06 Hz, 1 H). Example 2: 5-(2-amino-5H-pyrrolo[32-d]pyrimidin-4-yl)-24-dichlorophenol
Figure imgf000055_0001
The above compound was prepared as follows using General Procedure GB2 as described previously. 2-(2,4-Dichloro-5-methoxymethoxy-phenyl)-4,4,5,5-tetramethyl-
[1 ,3,2]dioxaborolane (compound A in General Procedure GD, 362 mg, 1 mmo!) was added to a solution of N'-(4-Chloro-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-N1N-dimethyl- formamidine (compound 2 in General Procedure GB2, 233 mg, 1 mmol) in 7 ml_ of 1 ,4- dioxane. The mixture was purged with N2 several times. Tetrakis (triphenylphosphine) palladium (115 mg, 0.1 mmol) was added, then Na2COa (1 ml_, 2M) was added to the mixture. The mixture was heated and stirred at 850C for 12 hours. H2O (20 ml_) was then added to the reaction mixture. EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil as the desired intermediate product (compound 3a: N'-[4-(2,4-dichloro- 5-hydroxy-phenyl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl]-N,N-dimethyl-formamidine)). This residue was used for next step reaction without further purification.
Figure imgf000055_0002
3a
Hydrogen chloride (4 mL, 16 mmol, 4M in dioxane) was added to a solution of compound 3a (426 mg, 1 mmol) in MeOH (3 mL). The reaction mixture (a clear light brown yellow solution) was stirred at room temperature for 12 hours. The solvent was evaporated and the residue was neutralized with saturated Na2COa. EtOAc (50 mL) and THF (50 mL) were added to the aqueous solution to extract. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil. The residue was purified by silica gel chromatography (gradient elution 0→10% MeOH in EtOAc) to give a white solid (95 mg, 31 % yield in two steps) as the desired final product. 1H NMR (400 MHz1 DMSOd6) δ ppm 5.96 (s, 2 H) 6.18 (dd, J=2.91 , 1.64 Hz, 1 H) 7.06 (s, 1 H) 7.53 (t, J=3.03 Hz, 1 H) 7.64 (s, 1 H) 10.78 (s, 1 H) 11.10 (s, 1 H). Example 3: 3-(2-amino-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-6-chloro-2-methylphenol
Figure imgf000056_0001
The above compound was prepared as follows using General Procedure GB2 as described previously. 2-(2,4-Dichloro-3-methoxymethoxy-phenyl)-4,4,5,5-tetramethyl- [1 ,3,2]dioxaborolane (compound B in General Procedure GD, 336 mg, 1 mmol) was added to a solution of N'-(4-Chloro-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-N,N-dimethyl- formamidine (compound 2 in General Procedure GB2, 233 mg, 1 mmol) in 7 mL of 1 ,4- dioxane. The mixture was purged with N2 several times. Tetrakis (triphenylphosphine) palladium (115 mg, 0.1 mmol) was added, then Na2COa (1 mL, 2M) was added to the mixture. The mixture was heated and stirred at 850C for 12 hours. H2O (20 mL) was added to the reaction mixture. EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil as the desired product. The residue was purified by silica gel chromatography (gradient elution 50→60% EtOAc in hexanes) to give the desired intermediate product (compound 3b: N'-[4-(2,4-Dichloro-3-methoxymethoxy-phenyl)-5H- pyrrolo[3,2-d]pyrimidin-2-yl]-N,N-dimethyl-formamidine)) as a white solid (36 mg, 9% yield).
Figure imgf000056_0002
Hydrogen chloride (0.3 mL, 1.2 mmol, 4M in dioxane) was added to a solution of compound 3b (35 mg, 0.09 mmol) in MeOH (3 mL). The reaction mixture (a clear light brown yellow solution) was stirred at room temperature for 12 hours. The solvent was evaporated and the residue was neutralized with saturated Na2CO3. EtOAc (20 ml_) and THF (20 mL) were added to the aqueous solution to extract. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil. The residue was purified by reversed phase chromatography to give a white solid (16 mg, 65% yield) as the desired final product.
1H NMR (400 MHz, DMSO-CZ6) δ ppm 2.28 (s, 3 H) 5.83 (s, 2 H) 5.83 (s, 2 H) 6.11 - 6.16 (m, 1 H) 6.13 - 6.17 (m, 1 H) 6.86 (d, J=7.58 Hz, 1 H) 7.18 (d, J=7.83 Hz, 1 H) 7.45 (t, J=2.91 Hz, 1 H) 9.21 - 9.42 (m, 1 H) 10.97 (s, 1 H). Example 4: 4-(2,4-dichloro-5-isopropoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-2- amine
Figure imgf000057_0001
The above compound was prepared as follows using General Procedure GB2 as described previously. 2-(2,4-dichloro-3-methoxymethoxy-phenyl)-4,4,5,5-tetramethyl- [1,3,2]dioxaborolane (compound D in General Procedure GD, 220 mg, 0.67 mmol) was added to a solution of N'-(4-chloro-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-N,N-dimethyl- formamidine (compound 2 in General Procedure GB2, 149 mg, 0.67 mmol) in 5 mL of 1,4-dioxane. The mixture was purged with N2 several times. Tetrakis (triphenylphosphine) palladium (77 mg, 0.07 mmol) was added, then Na2COa (1 rnL, 2M) was added to the mixture. The mixture was heated and stirred at 850C for 12 hours. H2O (20 mL) was added to the reaction mixture. EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil residue as the desired intermediate product (compound 3c: N1- [4-(2,4-dichloro-5-isopropoxy-phenyl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl]-N,N-dimethyl- formamidine)). The residue was used for the next step reaction without further purification.
Figure imgf000058_0001
SC
Hydrogen chloride (3 ml_, 12 mmol, 4M in dioxane) was added to a solution of compound 3c (0.67 mmol) in MeOH (3 mL). The reaction mixture (a clear light brown yellow solution) was stirred at room temperature for 12 hours. The solvent was evaporated and the residue was neutralized with saturated Na2CO3. EtOAc (2x50 mL) was added to the aqueous solution to extract. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil. The residue was purified by silica gel chromatography (gradient elution 70→80% EtOAc in hexanes) to give the desired product as a yellow oil (113.6 mg, 55% yield in two steps) as the desired final product. 1H NMR (400 MHz, DMSO-Cf6) δ ppm 1.28 (d, J=5.81 Hz, 6 H) 4.56 - 4.89 (m, 1
H) 5.89 (d, J=5.05 Hz, 1 H) 6.08 - 6.31 (m, 1 H) 7.11 - 7.33 (m, 1 H) 7.38 - 7.61 (m, 1 H) 7.67 - 7.88 (m, 1 H) 11.06 (s, 1 H).
Example 5: 4-(4-bromo-2-chloro-5-methoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-2- amine
Figure imgf000058_0002
The above compound was prepared as follows using General Procedure GB2 as described previously. 2-(4-Bromo-2-chloro-5-methoxy-phenyl)-4,4,5,5-tetramethyl- [1,3,2]dioxaborolane (from Combi-block, 275 mg, 1 mmol) was added to a solution of N'- (4-chloro-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-N,N-dimethyl-formamidine (compound 2 in General Procedure GB2, 350 mg, 1.6 mmol) in 7 mL of 1,4-dioxane. The mixture was purged with N2 several times. Tetrakis (triphenylphosphine) palladium (120 mg, 0.1 mmol) was added then Na2COa (1.5 mL, 2M) was added to the mixture. The mixture was heated and stirred at 850C for 12 hours. H2O (30 mL) was added to the reaction mixture. EtOAc (2x 100 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil as the desired intermediate product (compound 3d: (N'-[4-(4-bromo-2-chloro-5-methoxy- phenyl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl]-N1N-dimethyl-formamidine)). The residue was used for the next reaction step without further purification (582 mg).
Figure imgf000059_0001
3d
Hydrogen chloride (8 mL, 32 mmol, 4M in dioxane) was added to a solution of compound 3d (425 mmol, 1 mmol) in MeOH (5 mL). The reaction mixture (a clear light brown yellow solution) was stirred at room temperature for 12 hours. The solvent was evaporated and the residue was neutralized with saturated Na2CO3. EtOAc (2x100 mL) was added to the aqueous solution to extract. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil. The residue was purified by silica gel chromatography (gradient elution 80→90% EtOAc in hexanes) to give the desired final product as a yellow oil (268 mg, 72.8% yield in two steps). 1H NMR (400 MHz, DMSO-d6) δ ppm 3.86 (s, 3 H) 5.93 (s, 2 H) 6.17 (dd, J=3.03,
1.77 Hz, 1 H) 7.21 (s, 1 H) 7.53 (t, J=3.03 Hz, 1 H) 7.87 (s, 1 H) 11.08 (s, 1 H). Example 6: 4-(2,4-dimethoxy-6-methylphenyl)-5H-pyrrolo[3,2-d]pyrimidin-2-amine
Figure imgf000059_0002
The above compound was prepared as follows using General Procedure GB2 as described previously. 2-(2,4-Dimethoxy-6-methyl-phenyl)-4,4,5,5-tetramethyl-
[1,3,2]dioxaborolane (compound E in General Procedure GD, 120 mg, 0.43 mmol) was added to a solution of N'-(4-Chloro-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-N,N-dimethyl- formamidine (compound 2 in General Procedure GB2, 96.5 mg, 0.43 mmol) in 6 mL of 1 ,4-dioxane. The mixture was purged with N2 several times. Tetrakis (triphenylphosphine) palladium (50 mg, 0.043 mmol) was added, then Na2CO3 (0.65 ml_, 2M) was added to the mixture. The mixture was heated and stirred at 850C for 12 hours. H2O (10 ml_) was added to the reaction mixture. EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil residue as the desired intermediate product (compound 3e: (N'-[4-(2,4-dimethoxy-6-methyl-phenyl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl]- N,N-dimethyl-formamidine)). The residue was used for the next reaction step without further purification.
Figure imgf000060_0001
3e Hydrogen chloride (2 mL, 8 mmol, 4M in dioxane) was added to a solution of compound 3e (0.43 mmol) in MeOH (5 mL). The reaction mixture (a clear light brown yellow solution) was stirred at room temperature for 12 hours. The solvent was evaporated and the residue was neutralized with saturated Na2CO3. EtOAc (2x50 mL) was added to the aqueous solution to extract. The combined organic layer was dried, filtered, and concentrated to give a brown yellow oil. The residue was purified by reverse phase chromatography as a white solid (23.7 mg, 19.3% yield in two steps) as the desired final product.
1H NMR (400 MHz, DMSO-αk) δ ppm 2.02 (s, 3 H) 3.64 (s, 3 H) 3.82 (s, 3 H) 6.08 - 6.15 (m, 2 H) 6.17 (dd, J=2.78, 1.77 Hz, 1 H) 6.54 (d, J=1.26 Hz, 2 H) 7.48 (t, J=2.78 Hz, 1 H) 11.08 (s, 1 H).
Example 7: 4-[2,4-dichloro-5-(3,3,3-trifluoropropoxy)phenyl]-5H-pyrrolo[3,2- d]pyrimidin-2-amine
Figure imgf000060_0002
The above compound was prepared as follows using General Procedure GB3 as described previously. 3-bromo-1 ,1 ,1,-trifluoropropane (80 mg, cas#[460-32-2]) was added to a solution of the final compound in Example 2 (95 mg, 0.32 mmol) and NaH (10 mg, 0.6 mmol) in 3 mL of DMF under N2 atmosphere. The reaction mixture was stirred at 6O0C for 12 hours. H2O (50 mL) was added to the reaction mixture. EtOAc (100 mL) was added to extract the aqueous solution. The organic layer was dried, filtered, and concentrated to obtain a yellow oil. The residue was purified by silica gel chromatography (gradient elution 70→80% EtOAc in hexanes) to give a white solid (13 mg, 10% yield) as the desired final product. 1H NMR (400 MHz1 DMSO-c/6) δ ppm 2.72 - 2.95 (m, 2 H) 4.34 (t, J=5.68 Hz, 2 H)
5.93 (s, 2 H) 6.19 (dd, J=2.91, 1.64 Hz, 1 H) 7.36 (s, 1 H) 7.55 (t, J=3.03 Hz, 1 H) 7.78 (s, 1 H) 11.10 (s, 1 H).
Example 8: 4-{2,4-dichloro-5-[2-(diethylamino)ethoxy]phenyl}-5H-pyrrolo[3,2- d]pyrimidin-2-amine
Figure imgf000061_0001
The above compound was prepared as follows using General Procedure GB3 as described previously. Sodium hydride (15 mg, 0.4 mmol) was added to a solution of the final compound in Example 2 (41 mg, 0.14 mmol) in DMF. The mixture was stirred at room temperature for 30 minutes. Potassium carbonate (40 mg, 0.3 mmol) was added as a solid and then (2-Bromo-ethyl)-diethyl-amine hydrobromide (60 mg, 0.3 mmol) was added. The reaction was heated at 8O0C for 12 hours. H2O (50 mL) was added, EtOAc (50 mL) and THF (50 mL) were added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to obtain a brown yellow oil. The residue was purified by silica gel chromatography (gradient elution 50→60% MeOH in DCM) to give a colorless oil (17.7 mg, 26% yield) as the desired final product.
1H NMR (400 MHz1 DMSO-d6) δ ppm 0.96 (t, J=7.07 Hz, 6 H) 2.54 (q, J=7.07 Hz, 4 H) 2.81 (t, J=5.56 Hz, 2 H) 4.12 (t, J=5.68 Hz, 2 H) 5.92 (s, 2 H) 6.18 (dd, J=2.78, 1.77 Hz, 1 H) 7.28 (s, 1 H) 7.54 (t, J=3.03 Hz, 1 H) 7.74 (s, 1 H) 11.09 (s, 1 H). Example 9: 7-(2,4-dichlorophenyl)-2-methyl[1,3]thiazolo[5,4-d]pyrimidin-5-amine
Figure imgf000062_0001
The above compound was prepared as follows using General Procedure GC as described previously. /V-(7-chloro-2-methylthiazolo[5,4-c/]pyrimidin-5-yl)pivalamide (compound 4 in General Procedure GC) (112 mg, 0.39 mmol, 1.0 equiv.), 2,4- dichlorophenyl boronic acid (90 mg, 0.47 mmol, 1.2 equiv.) and Pd(PPh3J4 (45 mg, 0.039 mmol, 10 mol%) were dissolved in 4.5 ml_ dioxane under a nitrogen atmosphere. Cesium carbonate (192 mg, 0.59 mmol, 1.5 equiv.) in 0.5 mL water was added and the mixture was heated to reflux for 1 hour (under nitrogen). TLC showed complete consumption of the starting material and the mixture was concentrated in vacuo. The product was taken up in water (10 mL) and the product extracted with DCM (10 mL). The organic layer was dried (Na2SO4) and concentrated to dryness under reduced pressure. Purification by automated column chromatography (30 % EtOAc in heptane) yielded the desired intermediate product (compound 5a: Λ/-(7-(2,4-dichlorophenyl)-2- methylthiazolo[5,4-c/]pyrimidin-5-yl)pivalamide (130 mg, 0.34 mmol)) in 84 % yield.
Figure imgf000062_0002
5a
Compound 5a (115 mg, 0.29 mmol) was dissolved in methanol (4.0 mL) and 1 N NaOH (aq) (1.0 mL) was added. The mixture was stirred at 5O0C overnight and NMR showed complete removal of the pivaloyl group. The product was concentrated in vacuo, taken up in chloroform (50 mL) and washed with water. The organic layer was dried (Na2SO4), filtered and concentrated in vacuo to yield the desired final product (80 mg, 0.31 mmol) as a white solid in 87 % yield.
1H NMR (400 MHz, DMSO-d6) δ ppm 2.64 (s, 3 H) 7.09 (s, 2 H) 7.57 (d, J=1.26 Hz, 2 H) 7.79 (s, 1 H). Example 10: 7-(4-chloro-2-methylphenyl)-2-methyl[1 ,3]thiazolo[5,4-d]pyrimidin-5- amine
Figure imgf000063_0001
The above compound was prepared as follows using General Procedure GC as described previously. Λ/-(7-chloro-2-methylthiazolo[5,4-odpyrimidin-5-yl)pivalamide (compound 4 in General Procedure GC) (102 mg, 0.36 mmol, 1.0 equiv.), 2-chloro-4- methylphenyl boronic acid (73.2 mg, 0.43 mmol, 1.2 equiv.) and Pd(PPh3)4 (21.2 mg, 0.018 mmol, 5 mol%) were dissolved in 4.5 mL dioxane under a nitrogen atmosphere. Cesium carbonate (175 mg, 0.54 mmol, 1.5 equiv.) in 0.5 mL water was added and the mixture was heated to reflux for 1 hour (under nitrogen). TLC showed complete consumption of the starting material and the mixture was concentrated in vacuo. The product was taken up in water (10 mL) and the product extracted with dichloromethane (10 mL). The organic layer was dried (Na∑SO^ and concentrated to dryness under reduced pressure. Purification by automated column chromatography (20% EtOAc in heptane, ISCO) yielded the desired intermediate product (compound 5b: N-(7-(2-chloro- 4-methylphenyl)-2-methylthia2θlo[5,4-c(]pyrimidin-5-yl)pivalamide (120 mg, 0.32 mmol)) in 88% yield.
Figure imgf000063_0002
Compound 5b (120 mg, 0.32 mmol) was dissolved in methanol (4.0 mL) and 1 N NaOH (aqueous) (1.0 mL) was added. The mixture was stirred at 5O0C overnight and NMR showed complete removal of the pivaloyl group. The product was concentrated in vacuo, taken up in chloroform (50 mL) and washed with water. The organic layer was dried (Na2SO4), filtered and concentrated in vacuo, yielding the desired final product (90 mg, 0.31 mmol) as a white solid in 97% yield.
1H NMR (400 MHz, DMSO-d6) δ ppm 2.24 (s, 3 H) 2.65 (s, 3 H) 6.99 (s, 2 H) 7.30 - 7.40 (m, 1 H) 7.41 - 7.70 (m, 2 H). Example 11: /-(Z-chloro^-methylphenyl^-methylli^lthiazololδ.A-dlpyrimidin-δ- amine
Figure imgf000064_0001
The above compound was prepared as follows using General Procedure GC as described previously. N-(7-chloro-2-methylthiazolo[5,4-c(]pyrimidin-5-yl)pivalamide (compound 4 in General Procedure GC) (102 mg, 0.36 mmol, 1.0 equiv.), 2-chloro-4- methylphenyl boronic acid (73.2 mg, 0.43 mmol, 1.2 equiv.) and Pd(PPh3)4 (21.2 mg, 0.018 mmol, 5 mol%) were dissolved in 4.5 ml_ dioxane under a nitrogen atmosphere. Cesium carbonate (175 mg, 0.54 mmol, 1.5 equiv.) in 0.5 ml_ water was added and the mixture was heated to reflux for 1 hour (under nitrogen). TLC showed complete consumption of the starting material and the mixture was concentrated in vacuo. The product was taken up in water (10 mL) and the product extracted with dichloromethane (10 mL). The organic layer was dried (Na∑SCU) and concentrated to dryness under reduced pressure. Purification by automated column chromatography (20% EtOAc in heptane, ISCO) yielded the desired intermediate product (compound 5c: Λ/-(7-(2-chloro- 4-methylphenyl)-2-methylthiazolo[5,4-c/]pyrimidin-5-yl)pivalamide (120 mg, 0.32 mmol)) in 88% yield.
Figure imgf000064_0002
Compound 5c (120 mg, 0.32 mmo!) was dissolved in methanol (4.0 ml_) and 1 N NaOH (aqueous) (1.0 mL) was added. The mixture was stirred at 5O0C overnight and NMR showed complete removal of the pivaloyl group. The product was concentrated in vacuo, taken up in chloroform (50 mL) and washed with water. The organic layer was dried (Na2SO4), filtered and concentrated in vacuo to yield the desired final product (90 mg, 0.31 mmol) as a white solid in 97% yield.
1H NMR (400 MHz, DMSO-d6) δ ppm 2.39 (s, 3 H) 2.63 (s, 3 H) 7.02 (s, 2 H) 7.27 (d, J=7.83 Hz, 1 H) 7.35 - 7.49 (m, 2 H).
Example 12: 7-(4-bromo-2-chloro-5-methoxyphenyl)-2-methyl[1 ,3]thiazolo[5,4- d]pyrimidin-5-amine
Figure imgf000065_0001
The above compound was prepared as follows using General Procedure GC as described previously. A/-(7-chloro-2-methylthiazolo[5,4-d]pyrimidin-5-yl)pivalamide (compound 4 in General Procedure GC) (400 mg, 1.4 mmol, 1.0 equiv.), (4-bromo-2- chloro-5-methoxy)benzeneboronic acid (461 mg, 1.7 mmol, 1.2 equiv.) and Pd(PPh3)4 (162 mg, 0.14 mmol, 10 mol%) were dissolved in 4.5 mL dioxane under a nitrogen atmosphere. Cesium carbonate (690 mg, 2.11 mmol, 1.5 equiv.) in 2 mL water was added and the mixture was heated to 8O0C for 12 hours (under nitrogen). TLC showed complete consumption of the starting material and the mixture was concentrated in vacuo. The product was taken up in water (50 mL) and the product extracted with EtOAc (2x100 mL). The organic layer was dried (Na2SO4) and concentrated to dryness under reduced pressure to yield the desired intermediate product (compound 5d: N-[7- (4-bromo-2-chloro-5-methoxy-phenyl)-2-methyl-thiazolo[5,4-d]pyrimidin-5-yl]-2,2- dimethyl-propionamide) (640 mg). Compound 5d was used for the next reaction step without further purification.
Figure imgf000066_0001
Compound 5d (640 mg, 1.36 mmol) was dissolved in methanol (10 ml_) and 1 N NaOH (aqueous) (4.0 m!_) was added. The mixture was stirred at 550C overnight and NMR showed complete removal of the pivaloyl group. The product was concentrated in vacuo and neutralized with HOAc-NaOAc buffer. EtOAc (2x100 ml_) was added to extract the aqueous solution. The combined organic layer was dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by reversed phase HPLC to yield the desired final product (135 mg, 25.7% yield in two steps).
1H NMR (400 MHz, DMSO-d6) δ ppm 2.65 (s, 3 H) 3.85 (s, 3 H) 7.09 (s, 2 H) 7.27 (s, 1 H) 7.86 (s, 1 H).
Example 13: 4-[2-chloro-5-methoxy-4-(1-methyl-1 H-pyrazol-4-y!)phenyl]-5H- pyrrolo[3,2-d]pyrimidin-2-amine
Figure imgf000066_0002
The above compound was prepared as follows using General Procedure GE as described previously. 1-Methylpyrazole-4-boronic acid pinacol ester (38 mg, 0.18 mmol) was added to a solution of 4-(2,4-dichloro-5-isopropoxyphenyl)-5H-pyrrolo[3,2- d]pyrimidin-2-amine (final compound from Example 5) (60 mg, 0.15 mmol) in 5 mL of
1 ,4-dioxane. The mixture was purged with N2 several times. Tetrakis
(triphenylphosphine) palladium (17.4 mg, 0.015 mmol) was added then Na2CO3 (0.5 mL, 2M) was added to the mixture. The mixture was heated and stirred at 850C for 12 hours. H2O (30 mL) was added to the reaction mixture. EtOAc (2x20 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a yellow oil. DCM (10 mL) was added to this oil residue and a precipitate formed. The precipitate was collected and rinsed with DCM to give the desired final product (4.5 mg, 8.4% yield).
1H NMR (400 MHz1 DMSO-c/6) δ ppm 3.89 (s, 3 H) 3.90 (s, 3 H) 5.81 - 5.95 (m, 2 H) 6.18 (d, J=3.03 Hz, 1 H) 7.15 (s, 1 H) 7.51 (t, J=2..65 Hz, 1 H) 7.83 (s, 1 H) 8.06 (s, 1 H) 8.27 (s, 1 H) 11.08 (S1 1 H).
Table 1
Figure imgf000067_0002
Example 16: 7-[2,4-Dichloro-6-(2-pyrazol-1 -yl-ethoxy)-phenyl]-thiazolo[5,4- d]pyrimidin-5-ylamine.
Figure imgf000067_0001
Figure imgf000068_0001
Example 16
Synthesis of compound 16a: ethyl 2-amino-2-cvanoacetate:
Ethyl cyanoglyoxylate-2-oxime (50 g, 0.35 mol) was suspended in H2O (300 ml). Sat. aq. NaHCθ3 solution (150 ml) added in portions to the suspension and the suspension became a orange solution. Sodium dithionite (85% tech, 170 g, 0.98 mol) was scooped in portions. After the addition the mixture was stirred for 30 minutes and the temperature rose to 42 0C. The reaction mixture was them immediately extracted with CH3CI (4x 200 ml). The combined organic fraction was dried (Na2SO4), filtered and concentrated under reduced pressure affording a red/brown oil (34 g, 265 mmol, 76%). 1H-NMR (300 MHz, CDCI3) δ 4.42 (bs, 1 H), 4.30, (q, 2H, J = 7 Hz), 1.95 (bs, 2H), 1.31 (t, 3H, J = 7 Hz).
Synthesis of compound 16b: ethyl 2-cvano-2-formamidoacetate: Compound 16a (31 g, 0.24 mol) was dissolved in formic acid (310 ml). Acetic anhydride (34 ml, 36 mmol, 1.5 eq.) was added drop wise and after complete addition the mixture was stirred for 2 hours. The volatiles were removed under reduced pressure. The residue was tripped with toluene (2x 500 ml). The crude material (35 g) was purified by column chromatography (silica, heptanes-EtOAc 1/1). The product (yellow solid) was isolated in two fractions. The core fraction (9 g, 58 mmol, 24%) was pure material according to 1H NMR. 1H-NMR (300 MHz, CDCI3) δ 8.31 (s, 1H), 6.72 (bs, 1 H), 5.55 (d, 1 H, J = 6 Hz), 4.38 (q, 2H1 J = 7 Hz), 1.37 (t, 3H, J = 7 Hz).
Synthesis of compound 16c: ethyl 5-aminothiazole-4-carboxylate: Compound 16b (9.0 g, 58 mmol) was suspended in toluene (200 ml). Lawesson's reagent (23.0 g, 58 mmol, 1 eq.) was added in one portion. The obtained yellow suspension was stirred overnight at 70 0C under a N2 atmosphere. The reaction mixture was cooled to 0 0C and the toluene layer was decanted from the gummy material at the bottom of the of the flask and was extracted with 1 N. aq. HCI (6x 50 ml). The gummy material was dissolved in MeOH (150 ml) and concentrated under reduced pressure. The residue (brown oil) was suspended in TBME (300 ml) and was extracted with 1 N. aq. HCI (1x 300 ml, 2x 100 ml). The combined aqueous fraction was basified with 2 N. aq. NaOH (460 ml) and extracted with EtOAc (1x 300 ml, 2x 200 ml and 1x 100 ml). The combined organic layer was dried (Na2SO4), filtered and concentrated under reduced pressure affording the crude title compound (7 g). The crude material was purified by column chromatography (Silica, heptanes-EtOAc, 1/1) affording compound 16c (6 g, 35 mmol, 60 %) as a brown solid. 1H-NMR (300 MHz, CDCI3) δ 7.89 (s, 1H), 6.04 (bs,2H), 4.42 (q, 2H, J = 7 Hz), 1.43 (t, 3H, J = 7 Hz).
Synthesis of Compound 16d: 5-(3-Benzoyl-thioureidoMhiazole-4-carboxylic acid ethyl ester (Reference. Schenone et. al. Bioorganic. Med. Chem. Lett. 2004, 14, 2511-2517):
Benzoyl isothiocyanate (0.4ml, 3.02 mmol) was added to a solution of compound 16c (500 mg, 2.9 mmol) in dioxane (25 mL) at room temperature under nitrogen. The mixture was heated to reflux (about 1000C) for 12h. The reaction was monitored by LC/MS. The solvent was evaporated to afford a white solid residue, which was triturated with heptanes. The colorless solid was collected by filtration to afford compound 16d (830 mg, 2.48 mmol, 85% yield). 1H NMR (CDCI3); 1.51 (t, J = 7Hz1 3H), 1.84 (bs, 1 H), 4.02 (q, J = 7Hz , 2H), 7.58 (m, 1 H)1 7.68 (m, 1 H), 8.02 (m, 2H), 8.43 (bs, 1 H), 9.32 (bs, 1 H), 14.85 (bs, 1H). Synthesis of Compound 16e, 5-Thioxo-5T6-dihydro-4H-thiazolor5,4-dipyrirnidin-7-one: Sodium hydroxide (2.9mL, 1M, 2.9 mmol) was added to compound 16d (0.97g, 2.9mmol). The mixture was heated to 1000C for 1h. The reaction was monitored by LC/MS. The reaction mixture was acidified with glacial acetic acid and a precipitate formed. The precipitate was collected and washed with water, and dried in a vacuum oven to afford compound 16e (394 mg, 2.13 mmol, 73 % yield) as a colorless solid. 1H NMR (de-DMSO); 8.88 (s, 1 H), 12.61 (bs, 1H), 13.49 (bs, 1H).
Synthesis of Compound 16f: 5-Methylsulfanyl-6H-thiazolof5,4-dipyrimidin-7-one: (Reference. Ta-Shma et al, Tetrahedron 2006, 62, 5469-5473): 2.56g (13.9mmol) of Compound 16e was taken up in water (20ml). Sodium hydroxide (1.23g, 30.8mmol) was added followed by dimethyl sulfate (1.32ml, 13.9mmol), and the reaction was heated to 4O0C. A suspension was rapidly formed, and LC-MS after 15 minutes indicated the reaction is complete (> 90% mono-methylation). The reaction was cooled in an ice-bath, and the solid filtered, and washed with water. The solid was resuspended in water, sonicated and acidified with concentrated hydrochloric acid, and the solid collected. The initial filtrate was also acidified with concentrated hydrochloric acid, and the solid formed filtered, and washed with water. The two solids were combined, and dried overnight in the vacuum oven to yield Compound 16f (> 95% pure) as a colorless solid (1.76g, 64%). A second crop (0.24g), which contained 10-15% of the bis-methyl compound was isolated from the mother liquor. 1H NMR (de-DMSO); 2.61 (s, 3H), 9.05 (s, 1 H)1 13.23 (bs, 1 H).
Synthesis of Compound 16q: 7-Chloro-5-methylsulfanyl-thiazolor5,4-d1pyrimidine: Small Scale: Compound 16f (0.49g, 2.9mmol) was suspended in phosphorus oxychloride (10ml). The reaction was heated to 1000C, and after 15 minutes became homogeneous, and turned brown. The reaction was heated for 1 hour, and LC-MS indicated disappearance of starting material. The reaction was cooled, and the phosphours oxychloride removed in vacuo. Ice-water was added cautiously to the residue at O0C. The solution was basified with aqueous ammonia, and extracted with ethyl acetate. The organic extracts were dried over sodium sulfate, and the solvent removed. Trituration with heptanes afforded a brown solid, which was purified by chromatography on silica gel eluting with heptanes/ethyl acetate to afford Compound 16g (0.18g, 33%) as a colorless solid. 1H NMR (CDCI3); 2.72 (s, 3H), 8.99 (s, 1 H). CHN. CaIc. C 34.02%, H 2.16%. N 18.48%. Found (0.11eq. ethyl acetate), C 33.78%, H 2.14%, N 18.1%.
Large Scale: Compound 16f (1.76g, 10.4mmol) was suspended in phosphorus oxychloride (35ml). The reaction was heated to 10O0C, and after 15 minutes became homogeneous, and turned brown. The reaction was heated for 1 hour, and LC-MS indicated disappearance of starting material. The reaction was cooled, and the phosphours oxychloride removed in vacuo. Ice-water was added cautiously to the residue at O0C. The solution was basified with aqueous ammonia, and the brown solid formed isolated by filtration. The solid was taken up in ethyl acetate, dried over sodium sulfate, and the solution filtered. Removal of the solvent afford Compound 16g (1.19g, 62%) as a slightly yellow solid. 1H NMR (CDCI3); 2.72 (s, 3H), 8.99 (s, 1 H). Synthesis of compound 16h: 3,5-Dichloro-2-(5-methylsulfanyl-thiazolor5,4-d]pyrimidin-7- vD-phenol 2,4-dichloro-6-phenoxyboronic acid (compound F in GD-6, 178 rmg, 0.86 mmol) was added to a solution of compound 16g (170 mg, 0.78 mmol) in dioxane (10 ml_). The mixture was perged with N2 several times. Pd(PPh3)4 (54 mg, 0.05 mmol) was added then Na2Cθ3 (248 mg, 2.3 mmol, 1.2 mL, 2M in H2O) was added to the mixture. The mixture was heated at 120 °C in the microwave for 40 min. The reaction was cooled to room temperature and was poured into H2O (20 mL) to stir. EtOAc (2x 100 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown oil. The residue was purified using silica gel chromatography (gradient elution 45→55% EtOAc in hexanes) to afford the desired product as a white solid (200 mg, 74.4% yield). 1H NMR (400 MHz, DMSO-Cf6) δ ppm 2.62 (s, 3 H) 7.01 (d, J=1.77 Hz, 1 H) 7.22 (d, J=1.52 Hz, 1 H) 9.38 (s, 1 H) 10.72 (s, 1 H).
Synthesis of compound 16i: 7-("2,4-Dichloro-6-(2-pyrazol-1-yl-ethoxy)-phenvn-5- methylsulfanyl-thiazolor5,4-d1pyrimidine 1-(2-Bromo-ethyl)-1H-pyrazole (60 mg, 0.34 mmol) was added to a solution of compound 16h (110646-481 , 78 mg, 0.23 mmol) and K2CO3 (94 mg, 0.68 mmol) in DMF (3 mL). The reaction was heated at 120 0C in microwave for 40 min. H2O (20 mL) was added to the reaction mixture and EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a light brown oil (82 mg, 82% yield). This compound was used for the next step reaction without further purification.
Synthesis of compound 16i: 7-r2,4-Dichloro-6-(2-pyrazol-1-yl-ethoxy)-phenvπ-5- methanesulfonyl-thiazolor5,4-d]pyrimidine. M-Chloroperbenzoic acid (107 mg, 0.5 mmol, 2 equiv.) was added to a solution of 7- [2,4-Dichloro-6-(2-pyrazol-1-yl-ethoxy)-phenyl]-5-methylsulfanyl-thiazolo[5,4- d]pyrimidine (82 mg, 0.19 mmol, 1 equiv.) in DCM (10 mL) in an ice bath. The reaction was stirred at room temperature for 12h. Evaporate all the solvent to get yellow solid. The residue was stirred with 10%NaHSO3 (10 mL) and EtOAc (50 mL) was added to extract the aqueous solution. The organic layer was washed with NaOH (1M) and brine. The organic layer was dried, filtered, and concentrated to get a brown oil. This oil residue was used for the next step reaction without further purification. Synthesis of compound of Example 16:
Ammonia was bubbled through a solution of compound 16j (82 mg, 0.17 mmol) in dioxane (7 ml_) for 5 min. The mixture was stirred at 100 °C in a sealed tube for 12h. The mixture was evaporated to get a brown oil. The oil was purified by reverse phase chromatography (gradient elution 40→45% CH3CN in H2O (containing 0.1% HOAc)) (7 mg, 10% yield). 1H NMR (400 MHz, DMSO-d6) δ ppm 4.17 - 4.25 (m, 2 H) 4.24 - 4.33 (m, 2 H) 5.95 (t, J=Z 15 Hz, 1 H) 6.90 (d, J=2.27 Hz, 1 H) 7.13 (s, 2 H) 7.25 (d, J=1.77 Hz, 1 H) 7.28 - 7.31 (m, 1 H) 7.35 (d, J=1.77 Hz, 1 H) 8.83 (s, 1 H). Example 17: ethyl 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-5- hydroxythieno[2,3-d] pyrimidine-6-carboxylate
Figure imgf000072_0001
Synthesis of compound 17a: 4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carboxylic acid methyl ester: To a solution of 14.0 g of 2-methylthio-4,6-dichloropyrimidine (71.8 mmol) in 100 mL dry THF at -78 0C, 47 mL LDA (from Aldrich, 1.3 eq, 2.0 M in heptane/chlorobenzene/THF) was added slowly to give a deep brown solution. After being stirred at - 78 0C for 2.5 h, methyl chloroformate (8.32 mL, 1.5 eq) was added slowly. The resulting mixture was continued to stir at -78 0C for 30 min and was warmed slowly to 25 0C. The reaction was diluted with brine (500 mL) and extracted with CHCI3 (300 mL). The organic layer was separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was columned on silica gel using 4:1 heptane/EtOAc to afford 12.2 g of the desired product in 67 % yield as a yellow solid. 1H NMR (CDCI3): δ 2.58 (s, 3H), 3.98 (s, 3H); 13C NMR (CDCI3): δ 14.5, 53.5, 121.6, 158.0, 163.1 , 174.5.
Synthesis of compound 17b: 4-Chloro-6-ethoxycarbonylmethylsulfanyl-2-methylsulfanyl- pyrimidine-5-carboxylic acid methyl ester). To a pale yellow solution of 4,6-dichloro-2-methylsulfanyl-pyrimidine-5-carboxylic acid methyl ester (5.10 g, 20.1 mmol, 1.0 eq), ethyl thioglycolate (2.40 ml_, 21.9 mmol, 1.09 equiv), and EtOH (160 ml_) at 0 0C was added Et3N (2.90 ml_, 1.03 eq). The resulting yellow suspension was stirred at 0 0C for 20 min. Both TLC and crude 1H NMR indicated that small amounts of starting material were still left and di-displacement side product was generated. Therefore, another 0.20 ml. of ethyl thioglycolate (0.09 eq) and 0.47 mL of Et3N (0.17 eq) were added at 0 0C. After being stirred at 0 0C for additional 20 min, the solvent (EtOH) was removed under reduced pressure using water-bath (Caution: higher temperature may cause cyclization). The yellow residue was diluted with EtOAc (200 mL) and washed with brine (200 mL). The organic layer was collected, dried over Na2SO4l filtered, and concentrated under reduced pressure. The yellow oily residue was purified on silica gel (10:1 hexane/EtOAc to 3:1 hexane/EtOAc) to give 5.4 g of the desired product in 80 % yield as pale yellow oil. LC/MS: (APCI) 337 (M++1 ) 1H NMR (CDCI3): δ 1.28 (t, J = 5.49 Hz, 3H), 2.55 (s, 3H), 3.91 (s, 2H), 3.97 (s, 3H), 4.21 (q, J = 5.49 Hz, 2H). Synthesis of compound 17c: ethyl 4-(4-bromo-2-chloro-5-methoxyphenyl)-5-hydroxy-2- (methylthio)thienor2,3-dipyrimidine-6-carboxylate:
Sodium carbonate (472 mg, 4.5 mmol, 3 equiv.; 2M in H2O) was added to a solution of 4-Chloro-6-ethoxycarbonylmethylsulfanyl-2-methylsulfanyl-pyrimidine-5-carboxylic acid methyl ester (500 mg, 1.5 mmol, 1.0 equiv.) and (4-bromo-2-chloro-5- methoxy)benzeneboronic acid (354 mg, 1.3 mmol, 0.9 equiv.) in dioxane (10 mL). The mixture was perged with nitrogen several times and then Pd(PPh3)4 (172 mg, 0.15 mmol, 10 mol%) was added. The mixture was heated at 120C for 30 min in microwave. The reaction was monitored with LC/MS. The mixture was poured into water (20 ml) and the product extracted with EtOAc (2x100 ml). The organic layer was dried (Na2SO4), filtered, and concentrated to get a brown yellow oil. The residue was purified by silica gel chromatography (gradient elution 45→55% EtOAc in hexanes to give the desired product as a pale yellow solid (510 mg, 70% yield) as the desired product. 1H NMR (400 MHz1 CHLOROFORM-d) δ ppm 1.35 - 1.47 (m, 3 H) 2.69 (s, 3 H) 3.91 (s, 3 H) 4.35 - 4.49 (m, 2 H) 6.94 (s, 1 H) 7.69 (s, 1 H) 10.35 (s, 1 H).
Synthesis of compound 17d: ethyl 4-(4-bromo-2-chloro-5-methoxyphenyl)-5-hydroxy-2- (methylsulfonv0thienor2,3-d1pyrimidine-6-carboxylate. M-Chloroperbenzoic acid (464 mg, 2.1 mmol, 2 equiv.) was added to a solution of compound 17c (507 mg, 1 mmol, 1 equiv.) in DCM (15 ml_) in an ice bath. The reaction was stirred at room temperature for 12h. Evaporate all the solvent to get a pale yellow solid residue. The residue was stirred with 10%NaHSO3 (10 ml_) and EtOAc (100 ml_) was added to extract the aqueous solution. The organic layer was washed with NaOH (1 M) and brine. The organic layer was dried, filtered, and concentrated to get a yellow oil. This oil residue was used for the next step reaction without further purification. Synthesis of compound of Example 17.
Ammonia was bubbled through a solution of compound 17d (540 mg, 1 mmol) in dioxane (7 ml_) for 5 min. The mixture was stirred at 100 0C in a sealed tube for 12h. A yellow suspension was obtained. The mixture was pour into H2O (20 mL) and neutralized with HOAc-NaOAc buffer. EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a brown yellow oil. Methanol (20 mL) was added and the precipitate formed. The precipitate was collected and washed with more methanol to afford compound 5a (353 mg, 74.4% yield). 1H NMR (400 MHz, DMSO-cfe) δ ppm 1.25 (t, J=7.07 Hz, 3 H) 3.84 (s, 3 H) 4.26 (q, J=7.07 Hz, 2 H) 7.28 (s, 1 H) 7.54 - 7.68 (m, 2 H) 7.82 (s, 1 H) 10.25 (s, 1 H). Anal. Calcd for Ci6H13CIBrN3O4SO^SCH2CI2: C, 40.67; H, 2.84; N, 8.75. Found: C, 40.88; H, 2.97; N, 8.45. Example 18: ethyl 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-5- methoxythieno[2,3-d]pyrimidine-6-carboxylate.
Figure imgf000074_0001
lodomethane (43.3 mg, 0.31 mmol) was added to a solution of the compound of Example 17 (100 mg, 0.22 mol) and K2CO3 (151 mg, 1.1 mmol) in DMF (5 mL). The mixture was stirred at room temperature for 12h and monitored by LC/MS. The mixture was acidified with HOAc-NaOAc buffer and EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a yellow oil. The oil residue was purified by reverse phase HPLC (gradient elution 60→80% CH3CN in H2O (containing 0.1% HOAc)) to afford compound of Exmpale 18 as a pale yellow solid (8 mg, 8%yield). 1H NMR (400 MHz, DMSO-d6) δ ppm 1.27 (t, J=LOl Hz1 3 H) 3.52 (s, 3 H) 3.84 (s, 3 H) 4.20 - 4.32 (m, 2 H) 7.31 (s, 1 H) 7.54 (s, 2 H) 7.84 (s, 1 H).
Example 19: 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-5-methoxythieno[2,3- d]pyrimidine-6-carboxylic acid.
Figure imgf000075_0001
Lithium hydroxide (207 mg, 4.93 mmol) in H2O (3M, 1.6 mL) was added to a solution of compound of Example 18 (233 mg, 0.49 mmol) in dioxane (5 mL). The mixture was heated at 60 0C for 12h. The reaction was monitored by LC/MS. The mixture was acidified with HOAc-NaOAc buffer and EtOAc (2x100 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to afford the compound of Example 19 as a yellow solid (166 mg, 75.7% yield). 1H NMR (400 MHz, DMSO-Cf6) δ ppm 3.54 (s, 3 H) 3.84 (s, 3 H) 7.13 (s, 2 H) 7.23 (s, 1 H) 7.79 (s, 1 H). Example 20: 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-N-ethyl-5- methoxythieno[2,3-d]pyrimidine-6-carboxamide.
Figure imgf000075_0002
Ethylamine (1.3 mL, 1M in THF, 1.3 mmol) was added to a solution of compound of
Example 19 (56 mg, 0.13 mmole), diisopropylethylamine (100 mg, 0.8 mmol, 0.13 mL), and O-(7-azabenzotriazol-1-yl)-N,N)N',N'-tetramethyluronium phosphorus pentafloride (HATU) (53 mg, 0.14 mmol) in DMF (5 mL) under a nitrogen atmosphere. The reaction was allowed to stir at room temperature for 12 h. Saturated NaHCCb was added to the reaction mixture to quench the reaction. EtOAc (2x50 mL) was then added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a brown yellow oil. The oil residue was purified by silica gel chromatography (gradient elution 50→60% EtOAc in hexanes) to give the desired product as a pale yellow solid (18 mg, 30% yield). 1H NMR (300 MHz, DMSO-cfe) δ ppm 1.07 (t, J=7.06 Hz, 3 H) 3.17 (d, J=4.71 Hz, 2 H) 3.27 (s, 3 H) 3.85 (s, 3 H) 7.38 (s, 3 H) 7.79 - 8.04 (m, 2 H). Anal. Calcd for Ci7Hi6CIBrN4O3S: C, 43.28; H, 3.42; N, 11.88. Found: C, 42.88; H, 3.48; N, 11.48.
Example 21 : 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-N-ethyl-5- hydroxythieno[2,3-d]pyrimidine-6-carboxamide.
Figure imgf000076_0001
21 d ~ " Example 21 Synth esis of compound compound 21a: 4-(4-Bromo-2-chloro-5-methoxy-phenvO-5- ethoxymethoxy-2-methylsulfanyl-thienof2,3-dipyrimidine-6-carboxylic acid ethyl ester. Chloromethylethyl ether (210 mg, 2.2 mmol) was added to a solution of compound 17c (360 mg, 0.74 mmol) and DIEA (570 mg, 4.4 mmol) in DMF (5 mL) under nitrogen atmosphere. The reaction was stirred at room temperature for 12h and monitored by LC/MS. The reaction was poured into H2O (20 mL) and EtOAc (2x50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a light brown yellow oil (414 mg). The oil residue was used for the next step reaction without further purification. Synthesis of compound 21b: 4-(4-Bromo-2-chloro-5-methoxy-phenyl)-5-ethoxymethoxy- 2~methylsυlfanyl-thienof2,3-dipyrimidine-6-carboxylic acid.
Lithium hydroxide (317 mg, 2.5 mL, 3 M in H2O) was added to a solution of 4-(4-Bromo- 2-chloro-5-methoxy-phenyl)-5-ethoxymethoxy-2-methylsulfanyl-thieno[2,3-d]pyrimidine- 6-carboxylic acid ethyl ester in dioxane (5 mL). The mixture was heated at 60 0C for 12h and monitored by LC/MS. The reaction was neutralized with HOAc-NaOAc buffer and extracted with EtOAc (2x100 mL). The combined organic layer was dried, filtered, and concentrated to get a brown foam as the desired product (328 mg, 83.5% yield). Synthesis of compound 21c: 4-(4-Bromo-2-chloro-5-methoxy-phenyl)-5-ethoxymethoxy- 2-methylsulfanyl-thienof2,3-d]pyrirnidine-6-carboxylic acid ethylamide.
Ethylamine (198 mg, 4.4 mmol) was added to a solution of 4-(4-Bromo-2-chloro-5- methoxy-phenyl)-5-ethoxymethoxy-2-methylsulfanyl-thieno[2,3-d]pyrimidine-6-carboxylic acid (228 mg, 0.44 mmole), diisopropylethylamine (340 mg, 2.6 mmol, 0.5 mL), and O- (7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium phosphorus pentafloride (HATU) (250 mg, 0.66 mmol) in DMF (7 mL) under a nitrogen atmosphere. The reaction was allowed to stir at room temperature for 12 h. Saturated NaHCθ3 was added to the reaction mixture to quench the reaction. EtOAc (2x50 mL) was then added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to afford compound 21c as a brown oil (241 mg, 0.44 mmol). Compound 21c was used for the next step reaction without further purification.
Synthesis of compound 21 d: 4-(4-Bromo-2-chloro-5-methoxy-phenyl)-5-hydroxy-2- methylsulfanyl-thienolZS-dipyrimidine-θ-carboxylic acid ethylamide Hydrogen chloride (112 mg, 3.1 mmol, 0.8 mL, 4 M in dioxane) was added to compound 21c (241 mg, 0.44 mmol) in DCM (5 mL). The reaction mixture was stirred at room temperature for 12h and monitored by LC/MS. The reaction was evaporated to get a brown yellow oil residue as the desired product (215 mg, 0.44 mmol). This oil residue was used for the next step reaction without further purification.
Synthesis of compound of 21 e: 4-(4-Bromo-2-chloro-5-methoxy-phenyl)-5-hydroxy-2- methanesulfonyl-thienor2,3-d1pyrimidine-6-carboxylic acid ethylamide. M-Chloroperbenzoic acid (197 mg, 0.9 mmol, 2 equiv.) was added to a solution of 4-(4- Bromo^-chloro-δ-methoxy-phenyO-δ-hydroxy^-methylsulfanyl-thienop.S-dlpyrimidine- 6-carboxylic acid ethylamide (215 mg, 0.44 mmol, 1 equiv.) in DCM (15 mL) in an ice bath. The reaction was stirred at room temperature for 12h. Evaporate all the solvent to get a brown oil. The residue was stirred with 10%NaHSO3 (10 ml_) and EtOAc (100 ml_) was added to extract the aqueous solution. The organic layer was washed with NaOH (1M) and brine. The organic layer was dried, filtered, and concentrated to get a yellow oil. This oil residue was used for the next step reaction without further purification. Synthesis compound of Example 21 :
Ammonia was bubbled through a solution of 4-(4-Bromo-2-chloro-5-methoxy-phenyl)-5- hydroxy-2-methanesulfonyl-thieno[2,3-d]pyrimidine-6-carboxylic acid ethylamide (110646-383-1 , 215 mg, 0.44 mmol) in dioxane (7 ml_) for 5 min. The mixture was stirred at 100 °C in a sealed tube for 12h. The mixture was evaporated and purified by reverse phase HPLC to afford 2-amino-4-(4-bromo-2-chloro-5-methoxyphenyl)-N-ethyl- 5-hydroxythieno[2,3-d]pyrimidine-6-carboxamide as a white solid (20 mg, 10% yield). 1H NMR (400 MHz, DMSO-O6) δ ppm 1.04 - 1.17 (m, 3 H) 3.15 - 3.32 (m, 2 H) 3.76 - 3.93 (m, 3 H) 7.17 - 7.32 (m, 1 H) 7.46 (s, 1 H) 7.74 - 7.93 (m, 1 H) 8.35 (s, 2 H). Example 22: N-((1 R,5S)-3-aza-bicyclo[3.1.0]hexan-6-yl)-2-amino-4-(2,4-dimethoxy- 6-methylphenyl)thieno[2,3-d]pyrimidine-6- carboxamide.
Figure imgf000078_0001
223 2^ 22c Examples
Synthesis of compound 22a: 2-Amino-4-chloro-thienor2,3-diPyrimidine-6-carboxylic acid ethyl ester. Compound 22a was prepared and isolated as described in J. Heterocycl. Chem. (2005),
42(7), 1305-1310 (Tumkevicius, S. et al.).
Synthesis of compound 22b: ethyl 2-amino-4-(2τ4-d'ιmethoxy-6-methylphenv0thienof2,3- dlpyrimidine-6-carboxylate.
2-(2,4-dimethoxy-6-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (Compound E in General Prcedure D described previously, 178 mg, 0.9 mmol) was added to a solution of compound 22a (240 mg, 0.9 mmol) in dioxane (5 mL). The mixture was perged with N2 several times. Pd(PPh3)4 (120 mg, 0.1 mmol) was added then Na2CO3 (320 mg, 3 mmol, 1.5 mL, 2M in H2O) was added to the mixture. The mixture was heated at 80 0C for 12h. The reaction was cooled to room temperature and was poured into H2O (20 mL) to stir. EtOAc (2x 50 mL) was added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to give a brown oil. The residue was purified using silica gel chromatography (gradient elution 40→50% EtOAc in hexanes) to afford the desired product as a pale yellow solid (176 mg, 51.8% yield). 1H NMR (400 MHz, DMSO-Cf6) δ ppm 1.26 (t, J=7.07 Hz, 3 H) 2.00 (s, 3 H) 3.65 (S, 3 H) 3.82 (s, 3 H) 4.26 (q, J=7.07 Hz, 2 H) 6.55 (d, J=7.33 Hz, 2 H) 7.21 (s, 1 H) 7.32 (s, 2 H). Synthesis of compound 22c: 2-amino-4-(2,4-dimethoxy-6-methylphenyl)thienof2,3- dlDyrimidine-6-carboxylic acid.
Lithium hydroxide (320 mg, 7.5 mmol, 20 equiv., 2 M in H2O) was added to a solution of compound 22b in dioxane (5 mL). The mixture was stirred at 60 °C for 12h. The mixture was neutralized with HOAc-NaOAc buffer and extracted with EtOAc (2x50 mL). The combined organic layer was dried, filtered, and concentrated to get a pale yellow solid as the desired product. This compound was used for the next step reaction without further purification. 1H NMR (400 MHz, DMSO-cfe) δ ppm 1.96 (s, 3 H) 3.63 (s, 3 H) 3.81 (s, 3 H) 6.52 (d, J=4.55 Hz, 2 H) 6.64 (s, 1 H) 6.67 (s, 2 H) 12.73 (s, 1 H). Synthesis of compound of Example 22: 6-Amino-3-aza-bicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (synthesized based on reference: WO9824766 A1 19980611, 76.5 mg, 0.39 mmol) was added to a solution of compound 22c (111 mg, 0.32 mmole), 1, (3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (123 mg, 0.64 mmol), 4-methylmorpholine (162 mg, 1.6 mmol, 0.18 mL), and 1-hydroxy-benzotriazole (87mg, 0.64 mmol) in DMF (5 mL) under a nitrogen atmosphere. The reaction was allowed to stir at room temperature for 12 h. H2O (10 mL) was added to the reaction mixture to quench the reaction. EtOAc (2x50 mL) was then added to extract the aqueous solution. The combined organic layer was dried, filtered, and concentrated to get a brown yellow oil. Hydrogen chloride (4.8 mmol, 1.2 mL, 4M in dioxane) was added to a solution of the product (obtained above) in dioxane (5 mL). The reaction was stirred at room temperature for 12h. The solvent was evaporated to get a brown oil residue. The oil residue was purified by reverse phase chromatography (gradient elution 10→20% CH3CN in H2O (containing 0.1% HOAc)) to give the desired product as a white solid (25 mg, 18% yield). 1H NMR (400 MHz1 DMSO-de) δ ppm 1.68 (t, J=5.05 Hz, 2 H) 1.99 (s, 3 H) 2.62 - 2.78 (m, 1 H) 2.91 (dd, ./=11.24, 2.91 Hz, 2 H) 3.09 (d, J=11.37 Hz, 2 H) 3.64 (S, 3 H) 3.82 (s, 3 H) 6.55 (dd, J=9.47, 2.15 Hz, 2 H) 7.06 (s, 2 H) 7.39 (s, 1 H) 8.52 (d, j=4.04 Hz, 1 H). Anal. Calcd for C2iH23N5θ3S»2.25H2O: C, 54.12; H, 5.95; N, 15.03. Found: C, 54.39; H, 5.58; N, 14.73.
Example 23: ethyl 4-(2-(2-(1H-pyrazol-1-yl)ethoxy)-4,6-dichlorophenyl)-2- aminothieno[2,3-d]pyrimidine-6-carboxylate.
Figure imgf000080_0001
Example 23 Synthesis of compound 23a: ethyl 2-amino-4-(2,4-dichloro-6-hydroxyphenyl)thienor2,3- dipyrimidine-6-carboxylate.
Compound 23a was made following the same method that compound 22b was made.
1H NMR (400 MHz, DMSO-Gf6) δ ppm 1.27 (t, J=7.07 Hz, 3 H) 4.27 (q, J=7.07 Hz, 2 H)
7.01 (d, J=1.77 Hz, 1 H) 7.21 (d, ./=1.77 Hz, 1 H) 7.36 (s, 1 H) 7.43 (s, 2 H) 10.74 (s, 1 H). Anal. Calcd for Ci5HIiCI2N3O3S-I H2O: C, 44.79; H, 3.26; N, 10.45. Found: C,
44.42; H, 3.14; N, 10.15.
Synthesis of the compound of Example 23:
1-(2-Bromo-ethyi)-1H-pyrazole (123 mg, 0.7 mmol) was added to a solution of compound 23a (180 mg, 0.47 mmol) and K2CO3 (194 mg, 1.4 mmol) in DMF (10 mL). The reaction was heated at 120 °C in microwave for 40 min. H2O (20 mL) was added to the reaction mixture and EtOAc (2x50 mL) was added to extract the aqueous solution.
The combined organic layer was dried, filtered, and concentrated to get a colorless oil.
The oil was purified by silica gel chromatography (gradient elution 70→80% EtOAc in hexanes) to afford the desired product as a colorless oil (141.3 mg, 63% yield). 1H NMR (400 MHz, DMSO-CT6) δ ppm 1.28 (t, J=7.07 Hz, 3 H) 4.13 - 4.44 (m, 6 H) 5.88 (t, J=2.02
Hz, 1 H) 6.97 (d, J=2.27 Hz1 1 H) 7.18 (s, 1 H) 7.25 (d, J=1.52 Hz, 1 H) 7.31 (d, J=1.77
Hz, 1 H) 7.38 (d, J=1.77 Hz, 1 H) 7.42 (s, 2 H). Anal. Calcd for C2oHi7Cl2N5θ3S»0.25H2θ0.25hexanθs: C, 51.20; H, 4.20; N, 13.88. Found: C1 51.28; H1 4.00; N, 13.73. Examples 24-46: See Table 2.
Table 2
Figure imgf000081_0001
Figure imgf000082_0001
DMSO-d6) (m, 2 H) 4.18 J=4.67 Hz, 1 Hz, 1 H) 6.89 - 7.28 Hz, 1 J=1.77 Hz, 1
2 H) 1.78 4.16 (m, 1 H) Hz, 1 1 H)
Hz, 2 H) (d, 2 H) - 7.33 Hz, 2
(d, 2 H) 7.30 (d,
(q, (d, 2 H) - 7.41 1 H) Hz, 1 H)
2 H) (dd, 6.77 (d, 2 H) 7.35 (d,
Figure imgf000083_0001
Figure imgf000084_0001
Biological assay Example 1: HSP-90 Biochemical Assay
Compounds of the present invention were evaluated for potency against HSP-90 using a SPA (scintillation proximity assay) competition binding assay. Briefly, either full length or N-terminal HSP-90 that contains a 6-His tag on its C-terminus binds to copper on Yttrium-silicate scintillant beads via the His-tag. Tritiated propyl-Geldanamycin (pGA), whose structure is shown below, is an analog of a natural inhibitor of HSP-90 called Geldanamycin.
Figure imgf000085_0001
pGA
Tritiated pGA, which contains a tritiated propyl-amine group added at the #17 position, binds HSP-90 and brings the isotope into proximity with the beads. 17-n-propylamino- Geldanamycin can be prepared as described in U.S. Patent No. 4,261 ,989, which is incorporated herein by reference. A second tritiated compound that can also be used in this assay is shown below and is designated as Compound A.
Figure imgf000085_0002
Compound A
The "T" in the structure above indicates the position of the labeled tritiated hydrogen atoms. This compound has a Kd of 40 nM and can be prepared as follows.
Figure imgf000085_0003
The parent compound (/V-allyl-2-(5-chloro-2,4-dihydroxybenzoyl)isoindoline-1- carboxamide) shown above was first prepared as follows. Allylamine (2.5 ml_, 5 mmol, 2M in THF) was added to a solution of Boc(R,S)-1,3-dihydro-2H-isoindole carboxylic acid (263 mg, 1 mmole), diisopropylethy! amine (0.9 mL, 5 mmol), and O-(7- azabenzotriazol-1-yl)-N,N,N\N'-tetramethyluronium phosphorus pentafloride (HATU) (420 mg, 1.1 mmol) in 5 mL of DMF under a nitrogen atmosphere. The reaction was allowed to stir at room temperature for 12 hours. Saturated NaHCOa (30 mL) was added to the reaction mixture to quench the reaction. EtOAc (2x50 mL) was then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 was filtered off and the filtrate was evaporated to give a brown oil residue. The residue was purified by silica gel chromatography (gradient elution 40→50% EtOAc in hexanes) to give the desired intermediate product (321 mg, qunatitative yield) tert-butyl 1- [(allylamino)carbonyl]-1,3-dihydro-2/τ'-isoindole-2-carboxylate.
Hydrogen chloride (3 mL, 12 mmol; 4 M in dioxane) was added to a solution of te/t-butyl 1-[(allylamino)carbonyl]-1,3-dihydro-2H-isoindole-2-carboxylate (1 mmol) in DCM (5 mL) at room temperature. The reaction was heated and stirred at room temperature for 12 hours. The reaction mixture was evaporated to give an oil residue. The residue (Λ/-allylisoindoline-1-carboxamide) was used for the next step reaction without further purification. N-allylisoindoline-1-carboxamide (1 mmol) was then added to a solution of 5- chloro-2,4-bis(methoxymethoxy)benzoic acid (which can be prepared as shown in WO 2006/117669) (340 mg, 1.2 mmol), 4-methylmorpholine (2.2 mL, 20 mmol), N-(3- dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (460 mg, 2.4 mmol), and 1- hydroxy benzotriazole (330 mg, 2.4 mmol) in 12 mL of DMF under a nitrogen atmosphere. The reaction was allowed to stir at room temperature for 12 hours. H2O (50 mL) was added to the reaction mixture to quench the reaction. EtOAc (2 x 100 mL) was then added to extract the aqueous solution. Dry EtOAc layer over Na2SO4. The Na2SO4 was filtered off and the filtrate was evaporated to give a brown oil residue. The residue was purified by silica gel chromatography (gradient elution 50→60% EtOAc in hexanes) to give the desired intermediate product (423 mg, 91.8% yield) Λ/-allyl-2-[5- chloro-2,4-bis(methoxymethoxy)benzoyl]isoindoline-1-carboxamide. Hydrogen chloride (4 ml_, 16 mmol; 4 M in dioxane) was added to a solution of N- allyl-2-[5-chloro-2,4-bis(methoxymethoxy)benzoyl]isoindoline-1 -carboxamide (392 mg, 0.85 mmol) in DCM (5 ml_). The reaction was stirred at room temperature for 12 hours. The reaction mixture was neutralized with saturated NaHCO3 (aq) and then extracted with EtOAc (2 x 50 ml_). The combined organic layers were dried, filtered, and evaporated to give the desired final product as the parent compound (Λ/-allyl-2-(5- chloro-2,4-dihydroxybenzoyl)isoindoline-1 -carboxamide) as a white solid (221 mg, 69.7% yield). 1H NMR (400 MHz, DMSO-d6) δ ppm 3.57 (d, J=79.33 Hz, 2 H) 4.65 - 4.93 (m, 1 H) 4.97 - 5.19 (m, 1 H) 5.42 - 5.70 (m, 1 H) 5.68 - 5.95 (m, 1 H) 6.40 - 6.71 (m, 1 H) 6.92 (s, 1 H) 7.15 - 7.67 (m, 4 H) 8.28 (s, 1 H) 10.06 (s, 1 H) 10.40 (s, 1 H). Anal. Calcd for C19H17CIN2O4: C, 61.21; H, 4.60; N, 7.51. Found: C, 61.02; H, 4.63; N, 7.36.
Once the parent compound was made, Compound A was prepared using standard hydrogenation methods using tritium gas. The beta signal emitted from the isotope excites the scintillant, which creates a measurable signal. As competitive compounds are added to the assay mixture, they compete with bound tritiated pGA or Compound A at the ATP-binding site on the N- terminal of HSP-90. When a compound displaces the labeled pGA or Compond A, the signal is reduced (the beta-particles are no longer in proximity with the bead). This reduction in signal is used to quantify the extent to which the inhibitor/compound is competitive with pGA or Compound A.
The SPA assay for 3H-pGA (designated G1) and Compound A (designated G2) binding to HSP-90 was performed in 96-well flat bottom white plates (Corning #3604). For G1, typical reaction solutions contained 30 nM HSP-90 and 200 nM 3H-pGA in binding buffer (100 mM Hepes, pH 7.5 and 150 mM KCI). For G2, typical reaction solutions contained 5 nM HSP-90 and 50 nM of Compound A. For G1 , the 3H-pGA was first diluted to 33% label with unlabeled pGA that was synthesized and purified to give a final concentration of 200 nM. For G2, labeled Compound A was diluted with unlabeled Compound A to provide a ratio of labeledrunlabeled of 1 :2 for a final concentration of 50 nM. Inhibitors were added to the HSP-90/3H-pGA (or HSP-90/Compound A) solutions at eleven different concentrations for Ki determinations. The range of inhibitor concentrations were 100 μM, or an appropriate range, for solid samples and 10 μM for targeted library compounds and 4 mM liquid stocks. To determine percent inhibition, the compound was tested at 1 and 10 μM. The final DMSO in the samples was 4%. Copper-Ysi beads (Amersham, #RPNQ0096) that have been diluted in binding buffer were added to each well to give a final concentration of 100 μg/well. The plates were sealed, covered with a foil-covered lid and shaken for 30 minutes at room temperature. The beads were allowed to settle for 30 minutes after which the plates were counted using a Packard TopCount NXT instrument. This procedure has also been adapted for medium throughput using a Beckman Biomek FX. Samples were run in duplicate and on two separate days to assure an accurate value of Kj. For Ki determinations, the corrected cpm's (actual cpm's minus background) were plotted vs. inhibitor concentration using GraphPad Prism software. The data were fit to a generic IC50 equation, Y = Yl / (1 + [X]/IC5o), where Yl = Y-intercept and [X] is the competing ligand/inh/bitor. The IC50 was then used to calculate the Ki by using the Cheng-Prusoff equation:
Figure imgf000088_0001
Where cl = cold ligand concentration (varies), [hi] = concentration of hot ligand (200 nM or 50 nM) and Kd{hl} = 240 nM (for 3H-pGA) or 40 nM (for Compound A). Error was calculated as follows: IC50 error / IC50 value = fractional error and fractional error * K, value = Ki error.
In the cases in which inhibitor binds to HSP-90 so tightly that the population of free inhibitor molecules is significantly depleted by formation of the enzyme-inhibitor complex, the above equation is no longer valid. This is normally true when the observed
IC50 is about the same as the HSP-90 concentration. For a tight binding inhibitor, the following equation can be applied:
EL _ - (Kr + I0 -E0) + ^K^ + I0 -E0)2 + 4x Eo x Kr EL 2x £
Where K?p = K; x (1 + ^) EL and EL0 are the radioligand-HSP-90 complexes in the presence and absence of inhibitor, respectively. EUEL0 represents the fractional signal in the presence of inhibitor, lo, E0, and L0 are the inhibitor, HSP-90, and radioligand concentrations, respectively. Ki is the inhibition constant for the ligand, while KL is the binding affinity constant between the enzyme (HSP-90) and the ligand. Biological assay Example 2: HSP-90 Cellular Assay
Compounds of the present invention were evaluated for cell-based potency against HSP-90 using the Akt Luminex assay, which measures the turnover of the HSP- 90 client protein Akt1. NCI-H1299 cells (ATCC Number: CRL-5803) were treated with serial dilutions of HSP-90 compounds for 24 hours. Cell lysates were then assayed to measure the loss of Akt1 expression and thus determine cellular IC50 by Akt/PKB Beadmate (Upstate Catalog # 46-605) using a Luminex 100 system.
Table 2: HSP-90 Biochemical and cellular assay data for compounds shown in Examples 1 to 46.
Figure imgf000089_0001
Figure imgf000090_0001

Claims

ClaimsWe claim:
1. A compound of formula (II),
Figure imgf000091_0001
(II) wherein:
X is CR9 or N;
R9 is H, -OH, halogen, (C1 to C6) alky!, (Ci to C6) alkoxy or (Ci to C6) perfluoroalkyl;
R10 is selected from the group consisting of
(a) H1 (Ci to C6) perfluoroalkyl, (Ci to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl and (Ci to C6) alkoxy,
(b) (Ci to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl and (Ci to C6) alkoxy, all further substituted by 1-3 groups selected from halogen, -OH, cyano, -NH2, -NH-(Ci to
C3 alkyl), -N(Ci to C3 alkyl)2, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, phenyl and (C2 to C9) heteroaryl, wherein each (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, phenyl and (C2 to C9) heteroaryl is optionally further substituted by 1-3 groups selected from halogen and (Ci to C3 alkyl), (c) (C3 to C8 cycloalkyl), (C2 to C9) cycloheteroalkyl, phenyl and (C2 to C9) heteroaryl, all optionally further substitued by 1-3 groups selected from (Ci to C6) alkyl, halogen, -OH, cyano, (Ci to C6) alkoxy, -S(O)2-(Ci to C3 alkyl), -NH2, -NH-(Ci to C3 alkyl) and -N(Ci to C3alkyl)2) and
(d) -(CH2)P-C(O)R12, (e) -(CH2)p-C(O)-OR12, and
(f) -(CH2)p-C(O)NR13aR13b; each R11a , R11b and R11c is independently selected from the group consisting of (a) -OH, halogen, cyano, (Ci to C6) perfluoroalkyl, (Ci to C6) alkyl, (C2 to C6) alkenyl and (C2 to C6) alkynyl,
(b) (Ci to C6) alkyl, (C2 to C6) alkenyl and (C2 to C6) alkynyl, all further substituted by 1-3 groups selected from halogen, -OH, cyano, -S(O)2-(Ci to C3 alkyl), -NH2, -NH- (C1 to C3 alkyl), -N(Ci to C3 alkyl)2, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, phenyl and (C2 to Cg) heteroaryl, wherein each (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, phenyl and (C2 to Cg) heteroaryl is optionally further substituted by 1-3 groups selected from halogen and (Ci to C3 alkyl),
(c) (C3 to C3) cycloalkyl, (C2 to Cg) cycloheteroalkyl, all optionally further subsitued by 1-3 groups selected from (Ci to C6) alkyl, halogen, -OH, cyano, -S(O)2-(Ci to C3 alkyl), (C1 to C6) alkoxy, -NH2, -NH-(C1 to C3 alkyl) and -N(Ci to C3 alkyl)2,
(d) -C(O)-R15, -(C1 to C6 alkylene)-C(O)R1δ, -(C2 to C6 alkenylene)-C(O)R15 and - (C2 to C6 alkynylene)-C(O)R15,
(e) -0-R15, -O-(Ci to C6 alkylene)-R15, -0-(C2 to C6 alkenylene)-R15 and -0-(C2 to C6 alkynylene)-R15,
(f) -0-(C1 to C6 alkylene)-C(0)0R15, -0-(C2 to C6 alkenylene)-C(O)OR15 and -O- (C2 to C6 alkynylene)-C(O)OR15;
(g) -0-(C1 to C6 alkylene)-S(O)2R15, -0-(C2 to C6 alkenylene)-S(O)2R15 and -0-(C2 to C6 alkynylene)-S(O)2R15; (h) -0-(C^ to C6 alkylene)-C(O)NR16aR16b, -0-(C2 to C6 alkenylene)-
C(O)NR16aR16b and -0-(C2 to C6 alkynylene)-C(O) NR16aR16b, and
(i) -0-(C1 to C6 alkylene)-NR16aR16b, -0-(C2 to C6 alkenylene)-NR16aR16b and -O- (C2 to C6 alkynylene)-NR16aR16b; each R12 is independently H, (Ci to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl, (Ci to C6) perfluoroalkyl, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, (C6 to Ci4) aryl, (C2 to C9) heteroaryl, -(Ci to C3 alkylene)-(C3 to C8) cyclcoalkyl, -(Ci to C3 alkylene)- (C2 to Cg cycloheteroalkyl), -(Ci to C3 alkylene)- (C6 to C14 aryl) or -(Ci to C3 alkylene)- (C2 to Cg heteroaryl), wherein each R12 is optionally further substituted by 1-3 R14; each R13a and R13b is independently H, (C1 to C6) alkyl, (C2 to C6) alkenyl, (C2 to
C6) alkynyl, (Ci to C6) perfluoroalkyl, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, (C6 to Cu)aryl, (C2 to C9) heteroaryl, -(C1 to C3 alkylene)-(C3 to C8) cyclcoalkyl, -(C1 to C3 alkylene)- (C2 to Cg cycloheteroalkyl), -(Ci to C3 alkylene)- (C6 to Ci4 aryl) or -(Ci to C3 alkylene)- (C2 to C9 heteroaryl), each R13a or R13b is optionally further substituted by 1-3 R14; or, R13a and R13b, taken together with the nitrogen atom to which they are bound, form a (C2 to Cg) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C-2 to Cg) cycloheteroalkyl or (C2 to Cg) heteroaryl group is optionally further substituted by 1-3 R14; each R14 is independenly halogen, -OH, cyano, -S(O)2-(Ci to C3 alkyl), (Ci to C3) alkyl, (C1 to C3) perfluoroalkyl (Ci to C3) alkoxy, -C(O)-(CrC6 alkyl), -C(O)-O(Ci to C6 alkyl), -C(O)-NH2, -C(O)-NH(Ci to C6 alkyl) or -C(O)N(C1 to C6 alkyl)2; each R15 is independently H, (Ci to C6) alkyl, (C2 to C6) alkenyl, (C2 to C6) alkynyl,
(C1 to C6) perfluoroalkyl, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, (C6 to Ci4)aryl, (C2 to C9) heteroaryl, -(C1 to C3 alkylene)-(C3 to Ca) cyclcoalkyl, -(C1 to C3 alkylene)- (C2 to Cg cycloheteroalkyl), -(Ci to C3 alkylene)- (C6 to Cu aryl) or -(Ci to C3 alkylene)- (C2 to Cg heteroaryl), wherein each R15 is otpioanlly further substituted by 1-3 R17; each R16a and R16b is independently H, (C1 to CB) alkyl, (C2 to C6) alkenyl, (C2 to
C6) alkynyl, (C1 to C6) perfluoroalkyl, (C3 to C8) cycloalkyl, (C2 to C9) cycloheteroalkyl, (C6 to C14) aryl, (C2 to Cg) heteroaryl, -(C1 to C3 alkylene)-(C3 to C8) cyclcoalkyl, -(Ci to C3 alkylene)- (C2 to C9 cycloheteroalkyl), -(C1 to C3 alkylene)- (C6 to C14 aryl) or -(C1 to C3 alkylene)- (C2 to Cg heteroaryl), each R16a or R16b is optionally further substituted by 1-3 R17; or, R16a and R16b, taken together with the nitrogen atom to which they are bound, form a (C2 to Cg) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C2 to Cg) cycloheteroalkyl and (C2 to Cg) heteroaryl group is optionally further substituted by 1-3 R17; each R17 is independenly halogen, -OH, cyano, -S(O)2-(Ci to C3 alkyl), (C1 to C3) alkyl, (C1 to C3) perfluoroalkyl (Ci to C3) alkoxy, -C(O)-(Ci-C6 alkyl), -C(O)-O(C1 to C6 alkyl), -C(O)-NH2, -C(O)-NH(Ct to C6 alkyl) or -C(O)N(C1 to C6 alkyl)2; each p is independently 0, 1, 2 or 3; provided when X is CR9, R9 is H, and each R11a, R11b and R11c is unsubstituted (C1 to C6) alkyl, R10 is not -C(O)-OR12; or a pharmaceutically accpetalbe salt thereof.
2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein X is CR9.
3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R10 is -C(O)NR13aR13b.
4. The compound of claim 1 , or a pharmaceutically acceptable salt thereof, wherein R10 Is -C(O)-OR12.
5. The compound of claim 1 , or a pharmaceutically accpetalbe salt thereof, wherein R11a and R11b is independently halogen, Ci to C6 alkyl or C1 to C6 perfluoroalkyl, R11c is - 0-(C1 to C6 alkylene)-R15, -0-(C2 to C6 alkenylene)-R15 or -0-(C2 to C6 alkynylene)-R15, R15 is (C2 to C9) heteroaryl, (C3 to C8) cycloalkyl or (C2 to C9) heteroaryl, and R15 is optionally further substituted by 1-3 R17.
6. The compound of claim 1 , or a pharmaceutically acceptable salt thereof, wherein:
X is CR9 or N;
R9 is H, -OH, or (C1-C3 alkoxy); R10 is selected from (a) H, (Ci to C6) alkyl, and (C1 to C6) perfluoroalkyl,
(b) (Ci to C6) alkyl, further substituted by 1-3 groups selected from halogen and cyano,(c) (C3 to C8 cycloalkyl), (C2 to Cg) cycloheteroalkyl, phenyl and (C2 to C9) heteroaryl, all optionally further substitued by 1-3 groups selected from (C-i to C6) alkyl, halogen, -OH, cyano, (C1 to C6) alkoxy, -S(O)2-(Ci to C3 alkyl), -NH2, -NH-(C1 to C3 alkyl) and
Figure imgf000094_0001
and
(d) -(CH2)p-C(O)-OR12, wherein R12 is H or (Ci to C6) alkyl optionally substfituted by 1-3 halogen; and
(e) -(CH2)p-C(O)NR13aR13b, wherein each R13a and R13b is selected from H, (C1 to C6 alkyl) optionally substituted by 1-3 halogen, (C3 to Ca) cycloalkyl optionally substituted by 1-3 groups selected from halogen and (Ci to C3) alkyl, or R13a and R13b, taken together with the nitrogen atom to which they are bound, form a (C2 to Cg) cycloheteroalkyl group or a (C2 to Cg) heteroaryl group, wherein said (C2 to C9) cycloheteroalkyl or (C2 to C9) heteroaryl group is optionally further substituted by 1-3 groups selected from halogen, (C1 to C3) alkyl, (C1-C3) perfluoroalkyl and -S(O)2-(C1 to C3 alkyl); each R11a and R11b is independently halogen, -OH, (C1 to C6 alkyl), (C3 to C6) cycloalkyl or (C1 to C6) alkoxy; R11c is -0-(C1 to C6 alkylene)-R15, -0-(C2 to C6 alkenylene)-R15 or -0-(C2 to C6 alkynylene)-R15, R15 is (C2 to C9) heteroaryl, (C3 to C8) cycloalkyl or (C2 to C9) heteroaryl, and R15 is optionally further substituted by 1-3 R17.
7. The compound of claim 6, or a pharmaceutically acceptable salt thereof, wherein: R10 is -C(O)NR13aR13b, wherein R13a is H, R13b is (G, to C6 alkyl) optionally substituted by 1-3 halogen, or R13a and R13b, taken together with the nitrogen atom to which they are bound, form a (C2 to C9) cycloheteroalkyl group, the said (C2 to C9) cycloheteroalkyl group is optionally further substituted by 1-3 groups selected from halogen, (Ci to C3) alkyl, (Ci-C3) perfluoroalkyl and -S(O)2-(Ci to C3 alkyl); each R11a and R11b is independently halogen, (Ci to C6 alkyl), (Ci to C6) alkoxy;
R11c is -O-(Ci to C6 a(kylene)-R15 or -0-(C2 to C6 alkenylene)-R15, R15 is (C2 to C9) heteroaryl, and R15 is optionally further substituted by 1-3 groups seleted from halogen and (C1 to C3) alkyl.
8. A compound of the following structure,
Figure imgf000095_0001
wherein:
R1 is -(CH2)n-(C6 to C14) aryl, -(CH2)n-(C2 to C9) heteroaryl, or
-(CH2)π-C(O)NR4aR4b, wherein each of said (C6 to C14) aryl and (C2 to C9) heteroaryl is optionally substituted with at least one Rs group; R3 is H, (Ci to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, -(CH2)n-C(O)OR7 or
-(CH2)n-C(O)NR4aR4b;
R4a and R4b are each independently H, (C1 to C6) alkyl, (C2 to C8) alkenyl, (C2 to
C8) alkynyl, (C6 to C14) aryl, (C2 to C9) heteroaryl, (C2 to C9) cycloheteroalkyl, or (C3 to
Ca) cycloalkyl, all optionally substituted with at least one R8 group, or R4a and R4b, taken together with the nitrogen atom to which they are bound, form a (C2 to Cg) cycloheteroalkyl group, wherein said (C2 to C9) cycloheteroalkyl is optionally substituted with at least one R8 group; each R5 is independently -OH, (C1 to C6) alkyl, cyano, halogen, -C(O)NR4aR4b, -NR4aR4b, (C1 to C8) alkoxy, (C6 to Ci4) aryl or (C2 to C9) heteroaryl wherein each of said (Ci to C8) alkoxy, (C6 to Ci4) aryl, and (C2 to C9) heteroaryl is optionally substituted with at least one R6 group; each R6 is independently halogen, (Ci to C6) alkyl, (C6 to Cu) aryl, (C3 to C8) cycloalkyl, (C2 to C9) heteroaryl, (C2 to C9) cycloheteroalkyl, -(CH2)n-C(O)NR4aR4b, or -NR4aR4b;
R7 is H, or (C1 to C6) alkyl; each R8 is independently -OH, (Ci to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, (C-i to C8) heteroalkyl, or (C1 to C8) alkoxy; each n is independently 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof.
9. The compound or salt according to claim 8, wherein R1 is (C6 to C14) aryl or (C2 to C9) heteroaryl, wherein each of said (C6 to Ci4) aryl and (C2 to C9) heteroaryl is optionally substituted with at least one R5 group.
10. The compound or salt according to claim 9, wherein R1 is C6 aryl, wherein said C6 aryl is optionally substituted with at least one R5 group, R3 is -(CH2)n-C(O)NR4aR4b.
11. The compound or salt according to claim 10, wherein R3 is -C(0)NHR4b.
12. The compound or salt according to claim 8, wherein the compound of formula (I) has the following structure:
Figure imgf000096_0001
wherein:
R5a is (C1 to C8) alkoxy, which is optionally substituted with at least one R6 group;
R5b is halogen; and R5c is halogen.
13. A compound of the following structure,
Figure imgf000097_0001
wherein:
R2 is H, -(CH2V-(C6 to C14) aryl, -(CH2V(C2 to C9) heteroaryl, or - (CH2)n-C(O)NR4aR4b, wherein each of said (C6 to Ci4) aryl and (C2 to C9) heteroaryl is optionally substituted with at least one R5 group;
R3 is H, (C1 to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, -(CH2)n-C(O)OR7 or -(CH2)n-C(O)NR4aR4b;
R4a and R4b are each independently H, (C1 to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, (C6 to Ci4) aryl, (C2 to C9) heteroaryl, (C2 to C9) cycloheteroalkyl, or (C3 to C8) cycloalkyl, all optionally substituted with at least one R8 group, or R4a and R4b, taken together with the nitrogen atom to which they are bound, form a (C2 to Cg) cycloheteroalkyl group, wherein said (C2 to Cg) cycloheteroalkyl is optionally substituted with at least one R8 group; each R5 is independently -OH, (Ci to C6) alkyl, cyano, halogen, -C(O)NR4aR4b, -NR4aR4b, (Ci to C8) alkoxy, (C6 to Ci4) aryl or (C2 to C9) heteroaryl wherein each of said (C1 to C8) alkoxy, (C6 to C14) aryl, and (C2 to Cg) heteroaryl is optionally substituted with at least one R6 group;
R5a is (C1 to C8) alkoxy, which is optionally substituted with at least one R6 group;
Rδb is halogen; RSc is halogen; each R6 is independently halogen, (Ci to C6) alkyl, (C6 to Ci4) aryl, (C3 to C8) cycloalkyl, (C2 to C9) heteroaryl, (C2 to Cg) cycloheteroalkyl, -(CH2)n-C(O)NR4aR4b, or -NR4aR4b;
R7 is H, or (C1 to C6) alkyl; each R8 is independently -OH, (Ci to C6) alkyl, (C2 to C8) alkenyl, (C2 to C8) alkynyl, (Ci to C8) heteroalkyl, or (Ci to C8) alkoxy; each n is independently 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof.
14. The compound or salt according to claim 13, wherein R3 is -(CH2)n-C(O)NR4aR4b.
15. The compound or salt according to claim 14, wherein R3 is -C(O)NHR4b.
16. A pharmaceutical composition, comprising a compound or salt of claim 1.
17. A method of reducing abnormal cell growth in a mammal in need thereof, comprising the step of administering to said mammal a therapeutically effective amount of at least one compound or salt according to any one of claims 1 to 16.
18. The method of claim 17, wherein the abnormal cell growth is cancer.
19. The use of the compound of claim 1 , or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of cancer in a mammal.
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Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010103130A2 (en) 2009-03-13 2010-09-16 Katholieke Universiteit Leuven, K.U.Leuven R&D Novel bicyclic heterocycles
WO2011004132A1 (en) 2009-07-10 2011-01-13 Sanofi-Aventis Novel hsp90-inhibiting indole derivatives, compositions containing said derivatives, and use thereof
WO2011027081A2 (en) 2009-09-03 2011-03-10 Sanofi-Aventis Novel derivatives of 5,6,7,8-tetrahydroindolizine inhibiting hsp90, compositions containing same, and use thereof
WO2011093352A1 (en) 2010-01-27 2011-08-04 武田薬品工業株式会社 Thiazole derivative
WO2011147753A1 (en) 2010-05-26 2011-12-01 Katholieke Universiteit Leuven, K.U.Leuven R&D Antiviral activity of novel bicyclic heterocycles
WO2012035423A1 (en) 2010-09-15 2012-03-22 Katholieke Universiteit Leuven, K.U. Leuven R&D Anti-cancer activity of novel bicyclic heterocycles
WO2012130780A1 (en) * 2011-03-28 2012-10-04 F. Hoffmann-La Roche Ag Thiazolopyrimidine compounds
WO2013068458A1 (en) * 2011-11-10 2013-05-16 Ucb Pharma S.A. Therapeutically active thiazolo-pyrimidine derivatives
CN103443107A (en) * 2011-03-28 2013-12-11 弗·哈夫曼-拉罗切有限公司 Thiazolopyrimidine compounds
WO2015192119A1 (en) * 2014-06-13 2015-12-17 Yuma Therapeutics, Inc. Pyrimidine compounds and methods using the same
JP2016505025A (en) * 2013-01-18 2016-02-18 シャンハイ ケメックスプローラー カンパニー，リミティド Five-membered and six-membered heterocyclic compound and method for producing the same, pharmaceutical composition and use thereof
US9701691B2 (en) 2012-01-10 2017-07-11 Hoffman-La Roche Inc. Thienopyrimidine compounds
WO2018083288A1 (en) 2016-11-07 2018-05-11 Bayer Aktiengesellschaft Substituted sulfonyl amides for controlling animal pests
WO2021089673A1 (en) 2019-11-07 2021-05-14 Bayer Aktiengesellschaft Substituted sulfonyl amides for controlling animal pests
WO2021130259A1 (en) 2019-12-23 2021-07-01 UCB Biopharma SRL Dihydrocyclopenta-isoquinoline-sulfonamide derivatives compounds
WO2021130262A1 (en) 2019-12-23 2021-07-01 UCB Biopharma SRL Tetrahydrobenzo-quinoline sulfonamides derivative compounds
WO2022089449A1 (en) * 2020-10-26 2022-05-05 山东大学 PREPARATION OF SPECIFIC HEAT SHOCK PROTEIN 90α SUBTYPE INHIBITOR AND USE THEREOF

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001097979A (en) * 1999-07-28 2001-04-10 Takeda Chem Ind Ltd Condensed heterocyclic compound, and method of production and application thereof
WO2005021552A1 (en) * 2003-08-29 2005-03-10 Vernalis (Cambridge) Ltd Pyrimidothiophene compounds
WO2006008503A1 (en) * 2004-07-20 2006-01-26 Vernalis (Cambridge) Ltd Pyrimidothiophene compounds
WO2006090094A1 (en) * 2005-02-28 2006-08-31 Vernalis R & D Ltd Pyrimidothiophene compounds for use as hsp90 inhibitors
WO2007084560A2 (en) * 2006-01-17 2007-07-26 Signal Pharmaceuticals, Llc INHIBITORS OF TNFα, PDE4 AND B-RAF, COMPOSITIONS THEREOF AND METHODS OF USE THEREWITH

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001097979A (en) * 1999-07-28 2001-04-10 Takeda Chem Ind Ltd Condensed heterocyclic compound, and method of production and application thereof
WO2005021552A1 (en) * 2003-08-29 2005-03-10 Vernalis (Cambridge) Ltd Pyrimidothiophene compounds
WO2006008503A1 (en) * 2004-07-20 2006-01-26 Vernalis (Cambridge) Ltd Pyrimidothiophene compounds
WO2006090094A1 (en) * 2005-02-28 2006-08-31 Vernalis R & D Ltd Pyrimidothiophene compounds for use as hsp90 inhibitors
WO2007084560A2 (en) * 2006-01-17 2007-07-26 Signal Pharmaceuticals, Llc INHIBITORS OF TNFα, PDE4 AND B-RAF, COMPOSITIONS THEREOF AND METHODS OF USE THEREWITH

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
NICOLAUS B J R: "Symbiotic Approach to Drug Design" DECISION MAKING IN DRUG RESEARCH, XX, XX, 1 January 1983 (1983-01-01), pages 173-186, XP002197412 *

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US9096614B2 (en) 2011-11-10 2015-08-04 Ucb Pharma S.A. Therapeutically active thiazolo-pyrimidine derivatives
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