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SUBSTITUTED THIENOPYRIMIDINES THAT INTERACT WITH THE RAS SUPERFAMILY FOR THE TREATMENT OF CANCERS, INFLAMMATORY DISEASES, RASOPATHIES, AND FIBROTIC DISEASE

20230227466 · 2023-07-20

    Inventors

    Cpc classification

    International classification

    Abstract

    Provided herein are methods and compositions for treating cancers, inflammatory diseases, rasopathies, and fibrotic disease involving aberrant Ras superfamily signaling through the binding of compounds to the GTP binding domain of Ras superfamily proteins including, in certain cases, K-Ras and mutants thereof, and a method for assaying such compositions.

    Claims

    1. A compound of Formula IA: ##STR00369## or a pharmaceutically acceptable derivative thereof, wherein: —NR.sup.1R.sup.2 is ##STR00370## R.sup.3 is hydrogen, —CH.sub.3, —CF.sub.3, or phenyl; R.sup.4 is hydrogen, ##STR00371## and R.sup.5 is —CH.sub.3 or —CH.sub.2CF.sub.3.

    2. The compound or pharmaceutically acceptable derivative of claim 1, wherein —NR.sup.1R.sup.2 is: ##STR00372##

    3. The compound or pharmaceutically acceptable derivative of claim 1 or 2, wherein R.sup.3 is hydrogen.

    4. The compound or pharmaceutically acceptable derivative of claim 1 or 2, wherein R.sup.3 is —CH.sub.3.

    5. The compound or pharmaceutically acceptable derivative of claim 1 or 2, wherein R.sup.3 is phenyl.

    6. The compound or pharmaceutically acceptable derivative of claim 1, wherein: —NR.sup.1R.sup.2 is ##STR00373## R.sup.3 is hydrogen or phenyl; R.sup.4 is hydrogen or ##STR00374## and R.sup.5 is —CH.sub.3.

    7. The compound or pharmaceutically acceptable derivative of claim 6, wherein R.sup.3 is hydrogen.

    8. The compound or pharmaceutically acceptable derivative of claim 6, wherein R.sup.3 is phenyl.

    9. The compound or pharmaceutically acceptable derivative of any one of claims 1-8, wherein R.sup.4 is hydrogen.

    10. The compound or pharmaceutically acceptable derivative of any one of claims 1-8, wherein R.sup.4 is: ##STR00375##

    11. The compound or pharmaceutically acceptable derivative of any one of claims 1-10, wherein the compound of Formula IA is: ##STR00376##

    12. A compound of Formula IB: ##STR00377## or a pharmaceutically acceptable derivative thereof, wherein: R.sup.6 is ##STR00378## —NR.sup.7R.sup.8 is ##STR00379## R.sup.9 is hydrogen, —CH.sub.3, —CF.sub.3, or phenyl; and R.sup.10 is ##STR00380##

    13. The compound or pharmaceutically acceptable derivative of claim 12, wherein R.sup.6 is: ##STR00381##

    14. The compound or pharmaceutically acceptable derivative of claim 12, wherein R.sup.6 is: ##STR00382##

    15. The compound or pharmaceutically acceptable derivative of claim 12, wherein R.sup.6 is: ##STR00383##

    16. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00384##

    17. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00385##

    18. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00386##

    19. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00387##

    20. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00388##

    21. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00389##

    22. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00390##

    23. The compound or pharmaceutically acceptable derivative of any one of claims 12-15, wherein —NR.sup.7R.sup.8 is: ##STR00391##

    24. The compound or pharmaceutically acceptable derivative of any one of claims 12-23, wherein R.sup.9 is hydrogen.

    25. The compound or pharmaceutically acceptable derivative of any one of claims 12-23, wherein R.sup.9 is —CH.sub.3.

    26. The compound or pharmaceutically acceptable derivative of any one of claims 12-23, wherein R.sup.9 is phenyl.

    27. The compound or pharmaceutically acceptable derivative of any one of claims 12-26, wherein R.sup.10 is: ##STR00392##

    28. The compound or pharmaceutically acceptable derivative of any one of claims 12-26, wherein R.sup.10 is: ##STR00393##

    29. The compound or pharmaceutically acceptable derivative of any one of claims 12-26, wherein R.sup.10 is: ##STR00394##

    30. The compound or pharmaceutically acceptable derivative of any one of claims 12-26, wherein R.sup.10 is: ##STR00395##

    31. The compound or pharmaceutically acceptable derivative of any one of claims 12-30, wherein the compound of Formula IB is: ##STR00396## ##STR00397##

    32. The compound or pharmaceutically acceptable derivative of any one of claims 12-31, wherein the compound of Formula IB is: ##STR00398##

    33. The compound or pharmaceutically acceptable derivative of any one of claims 12-32, wherein the compound of Formula IB is: ##STR00399##

    34. The compound or pharmaceutically acceptable derivative of claim 12, wherein: R.sup.6 is ##STR00400## —NR.sup.7R.sup.8 is or ##STR00401## R.sup.9 is phenyl; and R.sup.10 is ##STR00402##

    35. The compound or pharmaceutically acceptable derivative of any one of claims 12-30 or claim 34, wherein —NR.sup.7R.sup.8 is: ##STR00403##

    36. The compound or pharmaceutically acceptable derivative of any one of claims 12-30 or claim 34, wherein —NR.sup.7R.sup.8 is: ##STR00404##

    37. The compound or pharmaceutically acceptable derivative of any one of claims 12-30 or claims 34-36, wherein R.sup.10 is: ##STR00405##

    38. The compound or pharmaceutically acceptable derivative of any one of claims 12-30 or claims 34-36, wherein R.sup.10 is: ##STR00406##

    39. The compound or pharmaceutically acceptable derivative of any one of claims 12-31 or claims 34-38, wherein the compound of Formula IB is: ##STR00407##

    40. A compound of Formula IC: ##STR00408## or a pharmaceutically acceptable derivative thereof, wherein: —NR.sup.11R.sup.12 is ##STR00409## ##STR00410## ##STR00411## and R.sup.13 is ##STR00412##

    41. The compound or pharmaceutically acceptable derivative of claim 40, wherein R.sup.13 is: ##STR00413##

    42. The compound or pharmaceutically acceptable derivative of claim 40, wherein R.sup.13 is: ##STR00414##

    43. The compound or pharmaceutically acceptable derivative of claim 40, wherein R.sup.13 is: ##STR00415##

    44. The compound or pharmaceutically acceptable derivative of any one of claims 40-43, wherein —NR.sup.11R.sup.12 is: ##STR00416##

    45. The compound or pharmaceutically acceptable derivative of any one of claims 40-44, wherein —NR.sup.11R.sup.12 is: ##STR00417##

    46. The compound or pharmaceutically acceptable derivative of any one of claims 40-44, wherein —NR.sup.11R.sup.12 is: ##STR00418##

    47. The compound or pharmaceutically acceptable derivative of any one of claims 40-44, wherein —NR.sup.11R.sup.12 is: ##STR00419##

    48. The compound or pharmaceutically acceptable derivative of any one of claims 40-44, wherein —NR.sup.11R.sup.12 is: ##STR00420##

    50. The compound or pharmaceutically acceptable derivative of any one of claims 40-44, wherein —NR.sup.11R.sup.12 is: ##STR00421##

    51. The compound or pharmaceutically acceptable derivative of any one of claims 40-43, wherein —NR.sup.11R.sup.12 is: ##STR00422## ##STR00423## ##STR00424##

    52. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00425##

    53. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00426##

    54. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00427##

    55. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00428##

    56. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00429##

    57. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00430##

    58. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00431##

    59. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00432##

    60. The compound or pharmaceutically acceptable derivative of any one of claims 40-43 or claim 51, wherein —NR.sup.11R.sup.12 is: ##STR00433##

    61. The compound or pharmaceutically acceptable derivative of any one of claims 40-60, wherein the compound of Formula IC is: ##STR00434## ##STR00435## ##STR00436## ##STR00437## ##STR00438## ##STR00439## ##STR00440## ##STR00441##

    62. The compound or pharmaceutically acceptable derivative of any one of claims 40-61, wherein the compound of Formula 1C is: ##STR00442## ##STR00443## ##STR00444##

    63. A compound of Formula ID: ##STR00445## or a pharmaceutically acceptable derivative thereof, wherein: —NR.sup.14R.sup.15 is ##STR00446##  and R.sup.16 is ##STR00447##

    64. The compound of claim 63, wherein —NR.sup.14R.sup.15 is: ##STR00448##

    65. The compound or pharmaceutically acceptable derivative of claim 63, wherein —NR.sup.14R.sup.15 is: ##STR00449##

    66. The compound or pharmaceutically acceptable derivative of claim 63, wherein —NR.sup.14R.sup.15 is: ##STR00450##

    67. The compound of claim 63, wherein —NR.sup.14R.sup.15 is: ##STR00451##

    68. The compound or pharmaceutically acceptable derivative of any one of claims 63-67, wherein R.sup.16 is: ##STR00452##

    69. The compound or pharmaceutically acceptable derivative of any one of claims 63-67, wherein R.sup.16 is: ##STR00453##

    70. The compound or pharmaceutically acceptable derivative of any one of claims 63-67, wherein R.sup.16 is: ##STR00454##

    71. The compound or pharmaceutically acceptable derivative of claim 63, wherein: —NR.sup.14R.sup.15 is ##STR00455## and R.sup.16 is ##STR00456##

    72. The compound or pharmaceutically acceptable derivative of any one of claims 63-71, wherein —NR.sup.14R.sup.15 is ##STR00457##

    73. The compound or pharmaceutically acceptable derivative of any one of claims 64-72, wherein the compound of Formula ID is: ##STR00458##

    74. A compound of Formula IE: ##STR00459## or a pharmaceutically acceptable derivative thereof, wherein: —NR.sup.17R.sup.18 is ##STR00460##

    75. The compound or pharmaceutically acceptable derivative of claim 74, wherein —NR.sup.17R.sup.18 is: ##STR00461##

    76. The compound or pharmaceutically acceptable derivative of claim 74, wherein —NR.sup.17R.sup.18 is: ##STR00462##

    77. The compound or pharmaceutically acceptable derivative of claim 74, wherein —NR.sup.17R.sup.18 is: ##STR00463##

    78. The compound or pharmaceutically acceptable derivative of claim 74, wherein —NR.sup.17R.sup.18 is: ##STR00464##

    79. The compound or pharmaceutically acceptable derivative of claim 74, wherein the compound of Formula IE is: ##STR00465##

    80. A compound of Formula IF: ##STR00466## or a pharmaceutically acceptable derivative thereof, wherein: —N.sup.19R.sup.20 is —NH.sub.2, ##STR00467## —NR.sup.21R.sup.22 is ##STR00468## and R.sup.23 is hydrogen or ##STR00469##

    81. The compound or pharmaceutically acceptable derivative of claim 80, wherein R.sup.23 is hydrogen.

    82. The compound or pharmaceutically acceptable derivative of claim 80, wherein R.sup.23 is: ##STR00470##

    83. The compound or pharmaceutically acceptable derivative of any one of claims 80-82, wherein the compound of Formula IF is: ##STR00471## ##STR00472##

    84. A compound of Formula IIA: ##STR00473## or a pharmaceutically acceptable derivative thereof, wherein: —NR.sup.24R.sup.25 is —NH.sub.2, ##STR00474##

    85. The compound or pharmaceutically acceptable derivative of claim 84, wherein the compound of Formula IIA is: ##STR00475## ##STR00476##

    86. A compound of Formula IIB: ##STR00477## or a pharmaceutically acceptable derivative thereof, wherein: R.sup.26 is ##STR00478## ##STR00479## R.sup.27 is hydrogen —CH.sub.3, or —CF.sub.3; and R.sup.28 is ##STR00480##

    87. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00481##

    88. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00482##

    89. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00483##

    90. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00484##

    91. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00485## ##STR00486##

    92. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00487##

    93. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00488##

    94. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00489##

    95. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00490##

    96. The compound or pharmaceutically acceptable derivative of claim 86, wherein R.sup.26 is: ##STR00491##

    97. The compound or pharmaceutically acceptable derivative of any one of claims 86-96, wherein R.sup.27 is hydrogen.

    98. The compound or pharmaceutically acceptable derivative of any one of claims 86-96, wherein R.sup.27 is —CH.sub.3.

    99. The compound or pharmaceutically acceptable derivative of any one of claims 86-96, wherein R.sup.27 is —CF.sub.3.

    100. The compound or pharmaceutically acceptable derivative of any one of any one of claims 86-98, wherein R.sup.28 is: ##STR00492##

    101. The compound or pharmaceutically acceptable derivative of any one of claims 86-98, wherein R.sup.28 is: ##STR00493##

    102. The compound or pharmaceutically acceptable derivative of claim 86, wherein: R.sup.26 is ##STR00494## R.sup.27 is hydrogen; and R.sup.28 is ##STR00495##

    103. The compound or pharmaceutically acceptable derivative of claim 102, wherein R.sup.26 is: ##STR00496##

    104. The compound or pharmaceutically acceptable derivative of claim 102, wherein R.sup.26 is: ##STR00497##

    105. The compound or pharmaceutically acceptable derivative of any one of claims 102-104, wherein R.sup.28 is: ##STR00498##

    106. The compound or pharmaceutically acceptable derivative of any one of claims 102-104, wherein R.sup.28 is: ##STR00499##

    107. The compound or pharmaceutically acceptable derivative of any one of claims 86-106, with the proviso that when: R.sup.26 is ##STR00500## and R.sup.27 is hydrogen; then R.sup.28 is not ##STR00501##

    108. The compound or pharmaceutically acceptable derivative of any one of claims 86-107, wherein the compound of Formula IIB is: ##STR00502## ##STR00503## ##STR00504##

    109. The compound or pharmaceutically acceptable derivative of any one of claims 86-107, wherein the compound of Formula IIB is: ##STR00505## ##STR00506## ##STR00507##

    110. The compound or pharmaceutically acceptable derivative of any one of claims 86-107, wherein the compound of Formula IIB is: ##STR00508##

    111. A compound or a pharmaceutically acceptable derivative thereof, wherein the compound is: ##STR00509##

    112. A compound of selected from the group consisting of compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, and 95, or a pharmaceutically acceptable derivative thereof.

    113. A compound of selected from the group consisting of compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, and 27, or a pharmaceutically acceptable derivative thereof.

    114. A compound of selected from the group consisting of compounds 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, and 39 or a pharmaceutically acceptable derivative thereof.

    115. A compound which binds to the GTP binding domain of one or more members of the Ras superfamily and inhibits the one or more members of the Ras superfamily with an IC.sub.50 value of less than 10 micromolar, wherein the compound is the compound or pharmaceutically acceptable derivative of any one of claims 1-114.

    116. The compound of claim 115, wherein one or more members of the Ras superfamily is Ras.

    117. The compound of claim 115, wherein one or more members of the Ras superfamily is Rho.

    118. The compound of claim 115, wherein one or more members of the Ras superfamily is Rac.

    119. The compound of claim 116, wherein the Ras is DIRAS1; DIRAS2; DIRAS3; ERAS; GEM; HRAS; KRAS; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS; or RRAS2.

    120. The compound of claim 119, wherein the Ras is HRAS, KRAS, NRAS, or a mutant thereof.

    121. The compound of claim 120, wherein the Ras is HRAS or a mutant thereof.

    122. The compound of claim 120, wherein the Ras is KRAS or a mutant thereof.

    123. The compound of claim 120, wherein the Ras is NRAS or a mutant thereof.

    124. The compound of claim 117, wherein the Rho is RHOA; RHOB; RHOBTB1; RHOBTB2; RHOBTB3; RHOC; RHOD; RHOF; RHOG; RHOH; RHOJ; RHOQ; RHOU; RHOV; RND1; RND2; RND3; RAC1; RAC2; RAC3; CDC42, or a mutant thereof.

    125. The compound of claim 117, wherein the Rho is Rac.

    126. The compound of claim 118 or 125, wherein the Rac is RAC1; RAC2; RAC3; RHOG, or a mutant thereof.

    127. The compound or pharmaceutically acceptable derivative of any one of claims 1-126, wherein the pharmaceutically acceptable derivative of the compound is a pharmaceutically acceptable salt of said compound.

    128. A method of inhibiting the function of one or more members of the Ras superfamily, comprising administering to a subject a compound which inhibits the one or more members of the Ras superfamily with an IC.sub.50 value of less than 10 μM, wherein the compound is the compound or pharmaceutically acceptable derivative of any one of claims 1-126 or the compound is the compound or pharmaceutically acceptable salt of claim 127.

    129. The method of claim 128, wherein one or more members of the Ras superfamily is Ras.

    130. The method of claim 128, wherein one or more members of the Ras superfamily is Rho.

    131. The method of claim 128, wherein one or more members of the Ras superfamily is Rac.

    132. The method of claim 128, wherein the Ras is DIRAS1; DIRAS2; DIRAS3; ERAS; GEM; HRAS; KRAS; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS; or RRAS2.

    133. The method of claim 132, wherein the Ras is HRAS, KRAS, NRAS or a mutant thereof.

    134. The method of claim 132, wherein the Ras is HRAS or a mutant thereof.

    135. The method of claim 132, wherein the Ras is KRAS or a mutant thereof.

    136. The method of claim 132, wherein the Ras is NRAS or a mutant thereof.

    137. The method of claim 130, wherein the Rho is RHOA; RHOB; RHOBTB1; RHOBTB2; RHOBTB3; RHOC; RHOD; RHOF; RHOG; RHOH; RHOJ; RHOQ; RHOU; RHOV; RND1; RND2; RND3; RAC1; RAC2; RAC3; CDC42, or a mutant thereof.

    138. The method of claim 137, wherein the Rho is Rac.

    139. The method of claim 131 or 138, wherein the Rac is RAC1; RAC2; RAC3; RHOG, or a mutant thereof.

    140. The method of claim 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of cancer.

    141. The method of any of claim 129 or 132-136, wherein the inhibiting the function of Ras is a treatment, prevention or amelioration of one or more symptoms of cancer.

    142. The method of any of claim 130 or 137-138, wherein the inhibiting the function of Rho is a treatment, prevention or amelioration of one or more symptoms of cancer.

    143. The method of any of claim 131 or 138-139, wherein the inhibiting the function of Rac is a treatment, prevention or amelioration of one or more symptoms of cancer.

    144. The method of any of claims 140-143, wherein the cancer is a solid tumor.

    145. The method of claim 144, wherein the solid tumor is hepatocellular carcinoma, prostate cancer, pancreatic cancer, lung cancer, breast cancer, ovarian cancer, colon cancer, small intestine cancer, biliary tract cancer, endometrium cancer, skin cancer (melanoma), cervix cancer, urinary tract cancer, or glioblastoma.

    146. The method of claim 145, wherein the solid tumor is pancreatic cancer.

    147. The method of claim 145, wherein the solid tumor is colon cancer.

    148. The method of claim 145, wherein the solid tumor is small intestine cancer.

    149. The method of claim 145, wherein the solid tumor is biliary tract cancer.

    150. The method of claim 145, wherein the solid tumor is endometrium cancer.

    151. The method of claim 145, wherein the solid tumor is lung cancer.

    152. The method of claim 145, wherein the solid tumor is breast cancer.

    153. The method of claim 145, wherein the solid tumor is skin cancer.

    154. The method of claim 145, wherein the solid tumor is cervix cancer.

    155. The method of claim 145, wherein the solid tumor is urinary tract cancer.

    156. The method of any of claims 140-143, wherein the cancer is a blood borne tumor.

    157. The method of claim 156, wherein the blood borne tumor is a leukemia.

    158. The method of claim 156, wherein the blood borne tumor is chronic lymphocytic leukemia (CLL), chronic myelocytic leukemia (CML), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), or acute myeloblastic leukemia (AML).

    159. The method of any one of claims 156-158, wherein the blood borne tumor is metastatic.

    160. The method of claim 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of an inflammatory disease.

    161. The method of any of claim 129 or 132-136, wherein inhibiting the function of Ras is a treatment, prevention or amelioration of one or more symptoms of an inflammatory disease.

    162. The method of any of claim 130 or 137-138, wherein the inhibiting the function of Rho is a treatment, prevention or amelioration of one or more symptoms of inflammatory disease.

    163. The method of any of claim 131 or 138-139, wherein the inhibiting the function of Rac is a treatment, prevention or amelioration of one or more symptoms of inflammatory disease.

    164. The method of any of claims 160-163, wherein the inflammatory disease is gastritis, schistosomiasis, cholangitis, chronic cholecystitis, pelvic inflammatory disease, chronic cervicitis, osteomyelitis, inflammatory bowel disease, reflux esophagitis, Barrett's esophagus, bladder inflammation (cystitis), asbestosis, silicosis, gingivitis, lichen planus, pancreatitis, protease mutation, lichen sclerosis, slaladenitis, bronchitis, Sjogren syndrome or Hashimoto's thyroiditis.

    165. The method of any of claims 160-163, wherein the inflammatory disease is Alzheimer's disease (AD), ankylosing spondylitis, arthritis (osteoarthritis, rheumatoid arthritis (RA), psoriatic arthritis), asthma, atherosclerosis, Crohn's disease, colitis, dermatitis, diverticulitis, fibromyalgia, hepatitis, irritable bowel syndrome (IBS), systemic lupus, erythematous (SLE), nephritis, Parkinson's disease, ulcerative colitis.

    166. The method of claim 165, wherein the inflammatory disease is Alzheimer's disease (AD).

    167. The method of claim 165, wherein the inflammatory disease is ankylosing spondylitis.

    168. The method of claim 165, wherein the inflammatory disease is arthritis (osteoarthritis, rheumatoid arthritis (RA), psoriatic arthritis).

    169. The method of claim 165, wherein the inflammatory disease is asthma.

    170. The method of claim 165, wherein the inflammatory disease is atherosclerosis.

    171. The method of claim 165, wherein the inflammatory disease is Crohn's disease.

    172. The method of claim 165, wherein the inflammatory disease is colitis.

    173. The method of claim 165, wherein the inflammatory disease is dermatitis.

    174. The method of claim 165, wherein the inflammatory disease is diverticulitis.

    175. The method of claim 165, wherein the inflammatory disease is fibromyalgia.

    176. The method of claim 165, wherein the inflammatory disease is hepatitis.

    177. The method of claim 165, wherein the inflammatory disease is irritable bowel syndrome (IBS).

    178. The method of claim 165, wherein the inflammatory disease is systemic lupus.

    179. The method of claim 165, wherein the inflammatory disease is erythematous (SLE).

    180. The method of claim 165, wherein the inflammatory disease is nephritis.

    181. The method of claim 165, wherein the inflammatory disease is Parkinson's disease.

    182. The method of claim 165, wherein the inflammatory disease is ulcerative colitis.

    183. The method of claim 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of a rasopathy.

    184. The method of any of claim 129 or 132-136, wherein the inhibiting the function of Ras is a treatment for a rasopathy.

    185. The method of any of claim 130 or 137-138, wherein the inhibiting the function of Rho is a treatment for a rasopathy.

    186. The method of any of claim 131 or 138-139, wherein the inhibiting the function of Rac is a treatment for a rasopathy.

    187. The method of any of claims 183-186, wherein the rasopathy is neurofibromatosis type 1, Noonan's syndrome or Costello syndrome.

    188. The method of any of claim 129 or 132-136, wherein the inhibiting the function of Ras is a treatment for Ras-associated autoimmune leukoproliferative disorder.

    189. The method of claim 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    190. The method of any of claim 129 or 132-136, wherein the inhibiting the function of Ras is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    191. The method of any of claim 130 or 137-138, wherein the inhibiting the function of Rho is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    192. The method of any of claim 130 or 137-138, wherein the inhibiting the function of Rac is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    193. The method of any one of claims 140, 160, 183, or 189, wherein one or more members of the Ras superfamily is Ras.

    194. The method of any one of claims 140, 160, 183, or 189, wherein one or more members of the Ras superfamily is Rho.

    195. The method of any one of claims 140, 160, 183, or 189, wherein one or more members of the Ras superfamily is Rac.

    196. A pharmaceutical composition, comprising the compound or pharmaceutically acceptable derivative of any one of claims 1-126, and a pharmaceutically acceptable carrier.

    197. The pharmaceutical composition of claim 196, wherein the pharmaceutical composition comprises a therapeutic amount of said compound or pharmaceutically acceptable derivative thereof.

    198. A pharmaceutical composition, comprising the compound or pharmaceutically acceptable salt of claim 127, and a pharmaceutically acceptable carrier.

    199. The pharmaceutical composition of claim 198, wherein the pharmaceutical composition comprises a therapeutic amount of said compound or pharmaceutically acceptable salt thereof.

    200. A method of inhibiting the function of one or more members of the Ras superfamily, comprising administering to a subject the pharmaceutical composition of any one of claims 196-199.

    201. A method of inhibiting the function of one or more members of the Ras superfamily, comprising administering to a subject the compound or pharmaceutically acceptable derivative of any one of claims 1-126.

    Description

    5. EXAMPLES

    Abbreviations

    [0639] PdXphosG3: (2-Dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate

    [0640] Pd(dppf)Cl.sub.2.Math.CH.sub.2Cl.sub.2: [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex with dichloromethane

    [0641] MTBE: Methyl tert-butyl ether

    [0642] HATU: 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate

    [0643] DIPEA: N,N-Diisopropylethylamine

    [0644] TCFH: N,N,N′,N′-tetramethylchloroformamidinium hexafluorophosphate

    [0645] The following examples are offered to illustrate but not to limit the disclosure.

    Example 1

    [0646] ##STR00137## ##STR00138##

    Rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenyl-6-(1H-pyrazol-3-yl)thieno[2,3-d]pyrimidin-4-amine (1)

    Step A: 2-(1-Methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol 1B

    [0647] Methyl 2-amino-4-phenylthiophene-3-carboxylate 1A (22.5 g, 96.45 mmol), 1-methyl-1H-imidazole-2-carbonitrile X1 (15.5 g, 144.67 mmol, 15.5 ml, 1.5 equiv), and potassium t-butoxide (86.58 g, 771.57 mmol) was dissolved in dry MeOH (220 mL) and stirred overnight at room temperature. The resulting mixture was evaporated and acetic acid was added to pH 5. Then it was extracted with EtOAc (250 mL), the organic layer was separated, dried over Na.sub.2SO.sub.4 and evaporated to give compound 1B, 2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol, (17.0 g, 55.13 mmol, 57.2% yield), which was used without additional purification in a further step.

    Step B: 6-Bromo-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol 1C

    [0648] Compound 1B, 2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol, (25.0 g, 81.07 mmol) was dissolved in DMF (250 mL), then N-bromosuccinimide (21.64 g, 121.61 mmol) was added at room temperature. Reaction mixture was heated at 65° C. overnight, cooled to room temperature and poured into ice (500 mL). The resulting precipitate was filtered and dried in air at 60° C. to give compound 1C, 6-bromo-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol, (20.0 g, 80.0% purity, 41.32 mmol, 51% yield) as yellow solid.

    Step C: 6-Bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 1D

    [0649] Compound 1C, 6-bromo-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol, (23.0 g, 59.4 mmol) was suspended in P(O)C13 (91.08 g, 594.03 mmol, 55.37 mL, 10.0 eq) and N,N-diisopropylethylamine (23.03 g, 178.21 mmol, 31.04 mL, 3.0 equiv) was added at room temperature. The reaction mixture was refluxed for 16 h, the solution was cooled to room temperature, evaporated under reduced pressure, poured in ice, diluted with ice-cold ammonia (200 mL, 20-25% of ammonia). The product was extracted with chloroform (2×500 mL) and evaporated. Compound 1D, 6-bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine, (22.0 g, 43.0% purity, 23.32 mmol, 39.3% yield) was obtained as yellow solid.

    Step D: rac-6-Bromo-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 1E

    [0650] Compound 1D, 6-bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine (4.46 g, 11.0 mmol) was dissolved in DMSO (50 mL) then N,N-diisopropylethylamine (3.55 g, 27.5 mmol, 4.79 ml, 2.5 equiv) and rac-(1R,3S)-3-methoxycyclopentan-1-amine (1.9 g, 16.5 mmol) was added in one portion. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. Compound 1E, rac-6-bromo-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine, was obtained as yellow viscous residue (1.25 g, 2.58 mmol, 23.5% yield).

    Step E: rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenyl-6-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)thieno[2,3-d]pyrimidin-4-amine 1F

    [0651] Compound 1E, rac-6-bromo-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine, (200.0 mg, 412.87 μmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[2-(trimethylsilyl)ethoxy]methyl-1H-pyrazole X2 (267.27 mg, 824.14 μmol), cesium carbonate (268.52 mg, 824.14 μmol) and PdXphosG3 (69.76 mg, 82.41 μmol) was mixed in dioxane/water (4 mL)/(0.2 mL). The reaction flask was filled with argon and the reaction mixture was stirred at 80° C. overnight, then cooled and filtered through a SiO.sub.2 pad. The mother liquid was concentrated and purified by HPLC. Compound 1F, rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenyl-6-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)thieno[2,3-d]pyrimidin-4-amine was obtained and used in the next step (57.5 mg, 66.0% purity, 63.06 μmol, 15.3% yield).

    Step F: rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenyl-6-(1H-pyrazol-3-yl)thieno[2,3-d]pyrimidin-4-amine (1)

    [0652] To the solution of compound 1F, rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenyl-6-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)thieno[2,3-d]pyrimidin-4-amine, (57.5 mg, 95.54 μmol) in CH.sub.2Cl.sub.2 (0.5 mL), TFA (0.5 mL) was added dropwise. This mixture was stirred 16 h at room temperature, concentrated and purified by HPLC. The Example compound (1), rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenyl-6-(1H-pyrazol-3-yl)thieno[2,3-d]pyrimidin-4-amine (6.1 mg, 92.0% purity, 11.9 μmol, 12.4% yield) was obtained.

    Example 2

    [0653] ##STR00139##

    N-((1s,3s)-3-methoxycyclobutyl)-2-(1-methyl-1H-imidazol-2-yl)-6-(1-methyl-1H-imidazol-4-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (2)

    Step A: 6-Bromo-N-((1s,3s)-3-methoxycyclobutyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 2A

    [0654] Compound 1D, 6-bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine, (13.76 g, 33.92 mmol) was dissolved in DMSO (140 mL) then N,N-diisopropylethylamine (10.96 g, 84.8 mmol, 14.77 mL, 2.5 equiv) and (1s,3s)-3-methoxycyclobutan-1-amine hydrochloride (7.0 g, 50.88 mmol) was added in one portion. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. Compound 2A, 6-bromo-N-((1s,3s)-3-methoxycyclobutyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine, (2.64 g, 91.0% purity, 5.11 mmol, 15.1% yield) was obtained as yellow viscous residue.

    Step B: N-((1s,3s)-3-methoxycyclobutyl)-2-(1-methyl-1H-imidazol-2-yl)-6-(1-methyl-1H-imidazol-4-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (2)

    [0655] Compound 2A, 6-bromo-N-((1s,3s)-3-methoxycyclobutyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine, (300.0 mg, 637.77 μmol), 1-methyl-4-(tributylstannyl)-1H-imidazole X3 (526.44 mg, 1.42 mmol), Pd(dppf)Cl.sub.2 CH.sub.2Cl.sub.2 (52.12 mg, 63.83 μmol), CuI (243.12 mg, 1.28 mmol), were dissolved in dioxane (5 mL). The reaction flask was filled with argon and the reaction mixture was stirred at 80° C. overnight. The resulting material was purified by HPLC without pre-workup. The Example compound, N-((1s,3s)-3-methoxycyclobutyl)-2-(1-methyl-1H-imidazol-2-yl)-6-(1-methyl-1H-imidazol-4-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (2), (6.6 mg, 14.0 μmol, 2.2% yield) was obtained.

    Example 3

    [0656] ##STR00140##

    Rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-6-(1-methyl-1H-imidazol-4-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (3)

    [0657] Compound 1E, rac-6-bromo-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (316.75 mg, 673.39 μmol), 1-methyl-4-(tributylstannyl)-1H-imidazole X3 (300.0 mg, 808.3 μmol), Pd(dppf)Cl.sub.2 CH.sub.2Cl.sub.2 (54.99 mg, 67.34 μmol), CuI (25.65 mg, 134.68 μmol), were dissolved in dioxane (3 mL). The reaction flask was filled with argon and the reaction mixture was stirred at 80° C. overnight. The resulting material was purified by HPLC without pre-workup. The Example compound, rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-6-(1-methyl-1H-imidazol-4-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (3), (6.1 mg, 92.0% purity, 11.56 μmol, 1.7% yield) was obtained.

    Example 4

    [0658] ##STR00141## ##STR00142##

    6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1S,3R)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (4)

    Step A: (1R,3S)-3-(Dibenzylamino)cyclopentan-1-ol 4B and (1S,3R)-3-(dibenzylamino)cyclopentan-1-ol 4C

    [0659] Sodium hydride (10.16 g, 423.56 mmol) was suspended in 100 mL of dry THF. The solution of rac-(1R,3S)-3-aminocyclopentan-1-ol (20.4 g, 201.7 mmol) in 200 mL of dry THE was added dropwise with water bath cooling. The resulting mixture was stirred until gas evolution ceased and then cooled to 0° C. (Bromomethyl)benzene (75.89 g, 443.73 mmol, 52.78 mL, 2.2 equiv) was added dropwise at that temperature, the resulting mixture was warmed to room temperature and then stirred overnight. After consumption of the starting material (.sup.1H-NMR control) the reaction mixture was poured into saturated aq. ammonium chloride solution (250 mL H.sub.2O+17.5 g ammonium chloride). The resulting mixture was extracted twice with 250 mL of CH.sub.2Cl.sub.2. Organic phases were combined, dried over sodium sulphate and evaporated to obtain a 1:1 mixture of (1R,3S)-3-(dibenzylamino)cyclopentan-1-ol 4B and (1S,3R)-3-(dibenzylamino)cyclopentan-1-ol 4C (35.0 g, 85.0% purity, 105.72 mmol, 52.4% yield). The product was purified by column chromatography to obtain 25 g (100% by LCMS) of a 1:1 mixture of (1R,3S)-3-(dibenzylamino)cyclopentan-1-ol 4B and (1S,3R)-3-(dibenzylamino)cyclopentan-1-ol 4C.

    [0660] Preparative Separation of Enantiomers 4B and 4C:

    [0661] Instrument: Agilent Technologies 1200: G1361A PrepPump, G2260A Prep ALS, G1315B DAD, G1364B Prer FC

    [0662] Column: Chiralcel OD-H (250*30 mm*5 m); Mobile phase: Hexane-IPA-MeOH, 95-2.5-2.5 Flow Rate: 12 mL/min; Column Temperature: 24° C.; Wavelength: 205 nm, 254 nm),

    [0663] RetTime (4B)=18.76 min; RetTime 4C)=34.49 min. The assignment of (1R,3S)-3-(dibenzylamino)cyclopentan-1-ol 4B and (1S,3R)-3-(dibenzylamino)cyclopentan-1-ol 4C absolute configuration was accomplished using Kazlauskas' Rule (see Step D and Step E below).

    Step B: (1S,3R)-N,N-dibenzyl-3-methoxycyclopentan-1-amine 4D and (1R,3S)-N,N-dibenzyl-3-methoxycyclopentan-1-amine 4E

    [0664] Sodium hydride (68.24 mg, 2.84 mmol, 2.0 equiv) was suspended in 5 mL of dry THF. The solution of (1S,3R)-3-(dibenzylamino)cyclopentan-1-ol 4C (400.0 mg, 1.42 mmol) in 4 mL of dry THE was added dropwise with water bath cooling. The resulting mixture was stirred until gas evolution ceased and then cooled to 0° C. Iodomethane (403.59 mg, 2.84 mmol) was added dropwise at that temperature, the resulting mixture was warmed to room temperature and then stirred overnight. After consumption of the starting material (.sup.1H-NMR control) the reaction mixture was poured into saturated aq. ammonium chloride solution (25 mL H.sub.2O+1.5 g ammonium chloride). The resulting mixture was extracted twice with 25 mL of CH.sub.2Cl.sub.2. Organic phases were combined, dried over sodium sulphate and evaporated to obtain (1R,3S)-N,N-dibenzyl-3-methoxycyclopentan-1-amine 4E (390.0 mg, 91.0% purity, 1.2 mmol, 84.5% yield). The product was used on the next step without additional purification. Using the same procedure but starting with (1R,3S)-3-(dibenzylamino)cyclopentan-1-ol 4B, gave (1S,3R)-N,N-dibenzyl-3-methoxycyclopentan-1-amine 4D (85% yield).

    Step C: (1S,3R)-3-methoxycyclopentan-1-amine 4F and (1R,3S)-3-methoxycyclopentan-1-amine 4G

    [0665] To a solution of (1R,3S)-N,N-dibenzyl-3-methoxycyclopentan-1-amine 4E (389.88 mg, 1.32 mmol) in MeOH (30 mL) was added palladium (70.0 mg, 657.77 μmol). The suspension was degassed and hydrogenated at 40° C. and ambient pressure for 7 days. Upon completion the mixture was cooled down to room temperature, the catalyst was removed off, and the filtrate was evaporated to dried to afford (1R,3S)-3-methoxycyclopentan-1-amine 4G (120.0 mg, 96.0% purity, 1.0 mmol, 75.8% yield). Using the same procedure but starting with (1S,3R)-N,N-dibenzyl-3-methoxycyclopentan-1-amine 4D gave (1S,3R)-3-methoxycyclopentan-1-amine 4F (77% yield)

    Step D: Enantiomeric resolution of (1R,3S)-3-(dibenzylamino)cyclopentan-1-ol 4B and (1S,3R)-3-(dibenzylamino)cyclopentan-1-ol 4C

    [0666] To the product of Step A prior to chiral separation (1:1 mixture of 4B and 4C) (502.0 mg, 1.78 mmol) and prop-1-en-2-yl acetate (536.31 mg, 5.36 mmol, 590.0 μl, 3.0 equiv) in 5 mL MTBE 0.5 g (10% mass %) of Lipase PS “Amano” IM (CAS [9001-62-1]) was added in one portion. Solution was stirred 72 h at room temperature. Solution was filtered off and concentrated. Residue was purified by column chromatography, eluted by Hex:EtOAc 10:1. Recovered alcohol 4C (270.0 mg, 959.52 μmol, 53.8% yield) was isolated as light-yellow oil. The stereochemical assignment of the acylated product (1R,3S)-3-(dibenzylamino)cyclopentyl acetate 4H (not isolated) was made using Kazlauskas' rule, “A rule to predict which enantiomer of a secondary alcohol reacts faster in reactions catalyzed by cholesterol esterase, lipase from Pseudomonas cepacia, and lipase from Candida rugosa” R. J. Kazlauskas, A. N. E. Weissfloch, A. T. Rappaport, and L. A. Cuccia, J. Org. Chem. 1991, 56, 2656-2665. Therefore 4C is (1S,3R)-3-(dibenzylamino)cyclopentan-1-ol.

    Step E: Comparison of .SUP.19.F NMR Measurements of R-Mosher's Acid Derivatives

    [0667] The .sup.19F NMR (in CDCl.sub.3) of the R-Mosher's ester prepared from the alcohol 4C isolated in Step D (compound 41, (1S,3R)-3-(dibenzylamino)cyclopentyl (R)-3,3,3-trifluoro-2-methoxy-2-phenylpropanoate, −72.06 ppm) matched the .sup.19F NMR of the R-Mosher's ester prepared from alcohol 4C isolated after chiral chromatography in Step A (−72.04 ppm). The .sup.19F NMR of a 1:1 mixture of R-Mosher's esters 4J (1R,3S)-3-(dibenzylamino)cyclopentyl (R)-3,3,3-trifluoro-2-methoxy-2-phenylpropanoate and 41 (1S,3R)-3-(dibenzylamino)cyclopentyl (R)-3,3,3-trifluoro-2-methoxy-2-phenylpropanoate (prepared from the 1:1 mixture of 4B and 4C from Step A prior to chiral separation) had peaks at −71.95 ppm (4J) and −72.05 ppm (4I). For the preparation and use of Mosher esters see “Methoxy-α-trifluoromethylphenylacetic acid, a versatile reagent for the determination of enantiomeric composition of alcohols and amines” J. A. Dale, D. L. Dull, H. S. Mosher, J. Org. Chem., 1969, 34(9), 2543-2549.

    Step F: 6-Bromo-N-((1S,3R)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 4K

    [0668] 6-Bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 1D (279.34 mg, 688.55 μmol), (1S,3R)-3-methoxycyclopentan-1-amine 4F (198.26 mg, 1.72 mmol) and N,N-diisopropylethylamine (267.12 mg, 2.07 mmol, 360.0 μL, 3.0 equiv) were mixed in DMSO (5 mL). The resulting reaction mixture was stirred at 100° C. for 15 h and then allowed to cool to room temperature. The mixture was diluted with water (10 mL) and extracted several times with EtOAc (15 mL*3). The combined extracts were washed with brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and the residue was purified by HPLC to afford the 6-bromo-N-((1S,3R)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 4K (119.0 mg, 245.66 μmol, 37.5% yield).

    Step G: 6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1S,3R)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (4)

    [0669] 6-Bromo-N-((1S,3R)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 4K (119.77 mg, 247.25 μmol), 1-isopropyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole X4 (70.06 mg, 296.7 μmol), cesium carbonate (161.12 mg, 494.49 μmol) and PdXphosG3 (20.93 mg, 24.72 μmol) was dissolved in degassed dioxane/H.sub.2O (2 mL:0.1 mL). The mixture was refluxed overnight. The mixture was cooled, filtered off, concentrated and purified by HPLC. The Example compound, 6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1S,3R)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (4) (6.7 mg, 13.04 μmol, 5.3% yield) was obtained.

    Example 5

    [0670] ##STR00143##

    6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (5)

    Step A: 6-Bromo-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 5A

    [0671] 6-Bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 1E (352.48 mg, 868.83 μmol), (1R,3S)-3-methoxycyclopentan-1-amine 4G (125.0 mg, 1.09 mmol) and N,N-diisopropylethylamine (336.87 mg, 2.61 mmol, 450.0 μL, 3.0 equiv) were mixed in DMSO (5 mL). The resulting reaction mixture was stirred at 100° C. for 15 h and then allowed to cool to room temperature. The mixture was diluted with water (10 mL) and extracted several times with EtOAc (15 mL*3). The combined extracts were washed with brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and the residue was purified by HPLC to afford the 6-bromo-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 5A (120.0 mg, 247.72 μmol, 30% yield).

    Step B: 6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (5)

    [0672] 6-Bromo-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine 5A (122.77 mg, 247.35 μmol), 1-isopropyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole X4 (70.06 mg, 296.7 μmol), cesium carbonate (161.12 mg, 494.49 μmol) and PdXphosG3 (20.93 mg, 24.72 μmol) was dissolved in degassed dioxane:H.sub.2O (2 mL:0.1 mL). Mixture was refluxed overnight. Mixture was cooled, filtered off, concentrated and purified by HPLC. The Example compound (5), 6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (29.0 mg, 95.0% purity, 53.64 μmol, 21.7% yield) was obtained.

    Example 6

    [0673] ##STR00144## ##STR00145##

    Rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (6)

    Step A: Ethyl 2-((tert-butoxycarbonyl)amino)-4-phenylthiophene-3-carboxylate 6B

    [0674] To a solution of ethyl 2-amino-4-phenylthiophene-3-carboxylate 6A (16.0 g, 64.7 mmol) and di-tert-butyl dicarbonate (16.94 g, 77.63 mmol) in dioxane (210 mL) N,N-dimethylpyridin-4-amine (790.37 mg, 6.47 mmol) was added. The mixture was stirred at 80° C. overnight. Then it was cooled, concentrated and purified by column chromatography. Eluent Hexane/EtOAc—10:1. Ethyl 2-[(tert-butoxy)carbonyl]amino-4-phenylthiophene-3-carboxylate 6B (15.0 g, 95.0% purity, 41.02 mmol, 63.4% yield) was obtained as white powder.

    Step B: Ethyl 5-bromo-2-[(tert-butoxy)carbonyl]amino-4-phenylthiophene-3-carboxylate 6C

    [0675] To a stirred solution of ethyl 2-[(tert-butoxy)carbonyl]amino-4-phenylthiophene-3-carboxylate 6B (15.0 g, 43.17 mmol) in DMF (30 mL) N-bromosuccinimide (9.22 g, 51.81 mmol) was added in one portion. The mixture was stirred at room temperature overnight. Then it was poured in ice-water (150 mL) and diluted with EtOAc (250 mL). Organic phase was washed with water (5×50 mL), dried over Na.sub.2SO.sub.4, concentrated and purified by column chromatography. Eluent Hex/EtOAc—7:1. Ethyl 5-bromo-2-[(tert-butoxy)carbonyl]amino-4-phenylthiophene-3-carboxylate 6C (10.5 g, 95.0% purity, 23.4 mmol, 54.2% yield) was obtained as light-yellow powder.

    Step C: Ethyl 2-[(tert-butoxy)carbonyl]amino-5-(1-methyl-1H-pyrazol-3-yl)-4-phenylthiophene-3-carboxylate 6D

    [0676] To a stirred suspension of ethyl 5-bromo-2-[(tert-butoxy)carbonyl]amino-4-phenylthiophene-3-carboxylate 6C (3.7 g, 8.68 mmol), 1-methyl-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole X5 (2.17 g, 10.41 mmol) and Cs.sub.2CO.sub.3 (5.65 g, 17.36 mmol) in dioxane/H.sub.2O (74 mL)/(3.7 mL) Pd(dppf)Cl.sub.2.Math.CH.sub.2Cl.sub.2 (634.98 mg, 867.8 μmol) was added in one portion under Ar. The mixture was stirred at 60° C. overnight. After cooling to room temperature, insolubles were filtered off and washed once with EtOAc (25 mL). The filtrate was concentrated and purified by column chromatography. Eluent Hex/EtOAc—100:1 to 10:1. Ethyl 2-[(tert-butoxy)carbonyl]amino-5-(1-methyl-1H-pyrazol-3-yl)-4-phenylthiophene-3-carboxylate 6D (1.9 g, 95.0% purity, 4.22 mmol, 48.7% yield) was obtained as yellow powder.

    Step D: Ethyl 2-amino-5-(1-methyl-1H-pyrazol-3-yl)-4-phenylthiophene-3-carboxylate 6E

    [0677] To a solution of ethyl 2-[(tert-butoxy)carbonyl]amino-5-(1-methyl-1H-pyrazol-3-yl)-4-phenylthiophene-3-carboxylate 6D (1.9 g, 4.44 mmol) in CH.sub.2Cl.sub.2 (20 mL) dioxane*HCl was added (7 mL 8M). Mixture was stirred overnight at room temperature. Ethyl 2-amino-5-(1-methyl-1H-pyrazol-3-yl)-4-phenylthiophene-3-carboxylate 6E (1.1 g, 3.36 mmol, 75.6% yield) was obtained as black oil.

    Step E: 2-(1-(2-Methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol 6F

    [0678] Ethyl 2-amino-5-(1-methyl-1H-pyrazol-3-yl)-4-phenylthiophene-3-carboxylate 6E (3.45 g, 11.02 mmol), 1-(2-methoxyethyl)-1H-imidazole-2-carbonitrile (2.0 g, 13.23 mmol), potassium t-butoxide (3.71 g, 33.07 mmol) was dissolved in dry MeOH (10 mL) and stirred overnight at room temperature. The residue was evaporated and acetic acid was added to pH 5. The resulted mixture was extracted with EtOAc (25 mL), organic layer was dried over Na.sub.2SO.sub.4, evaporated to give: 2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol 6F (4.2 g, 43.0% purity, 4.18 mmol, 37.9% yield) which was used without additional purification on the next step.

    Step F: 4-Chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidine 6G

    [0679] To the suspension of 2-[1-(2-methoxyethyl)-1H-imidazol-2-yl]-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-ol 6F (123.72 mg, 286.07 μmol) in phosphoroyl trichloride (438.63 mg, 2.86 mmol, 270.0 μL, 10.0 equiv) N,N-diisopropylethylamine (111.3 mg, 861.17 μmol, 150.0 μL, 3.0 equiv) was added at room temperature. The reaction mixture was refluxed for 16 h, cooled to room temperature and evaporated under reduced pressure. The residue was poured in ice and diluted with ice-cold ammonia (20 mL, 20-25% of ammonia), the product was extracted with CHCl.sub.3 (2×50 mL) and evaporated. 4-Chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidine 6G (125.0 mg, 75.4% purity, 209.01 μmol, 73.1% yield) was obtained as yellow solid.

    Step G: rac-N-((1R,3S)-3-Methoxycyclopentyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (6)

    [0680] To the solution of 4-chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidine 6G (146.44 mg, 324.74 μmol) in DMSO (5 mL), N,N-diisopropylethylamine (126.14 mg, 975.99 μmol, 170.0 μl, 3.0 equiv) and rac-(1R,3S)-3-methoxycyclopentan-1-amine (56.1 mg, 487.11 μmol) were added in one portion. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. The Example compound (6), rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (7.2 mg, 13.59 μmol, 4.2% yield) was obtained as yellow viscous residue.

    Example 7

    [0681] ##STR00146##

    N-((1s,3s)-3-Methoxycyclobutyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (7)

    [0682] To the solution of 4-chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidine 6G (138.18 mg, 306.42 μmol) in DMSO (5 mL), N,N-diisopropylethylamine (118.72 mg, 918.58 μmol, 160.0 μL, 3.0 equiv) with (1s,3s)-3-methoxycyclobutan-1-amine hydrochloride (46.49 mg, 459.63 μmol) were added in one portion. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. The Example compound (7), N-((1s,3s)-3-methoxycyclobutyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-6-(1-methyl-1H-pyrazol-3-yl)-5-phenylthieno[2,3-d]pyrimidin-4-amine (25.0 mg, 48.48 μmol, 15.8% yield) was obtained as yellow viscous residue.

    Example 8

    [0683] ##STR00147##

    Rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (8)

    Step A: 2-(1-(2-Methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 8B

    [0684] Methyl 2-aminothiophene-3-carboxylate 8A (415.85 mg, 2.65 mmol), 1-(2-methoxyethyl)-1H-imidazole-2-carbonitrile (600.0 mg, 3.97 mmol), potassium t-butoxide (890.58 mg, 7.94 mmol) was dissolved in dry MeOH (6 mL) and stirred overnight at room temperature. The reaction mixture was evaporated and acetic acid was added to pH 5. The resulted mixture was extracted with EtOAc (15 ml), organic layer was separated, dried over Na.sub.2SO.sub.4 and evaporated to give 2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 8B (850.0 mg, 73.8% purity, 2.27 mmol, 85.8% yield) which was used without additional purification on a further step.

    Step B. 4-Chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 8C

    [0685] To a suspension of 2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 8B (999.29 mg, 3.62 mmol) in phosphoroyl trichloride (5.55 g, 36.17 mmol, 3.37 mL, 10.0 equiv) N,N-diisopropylethylamine (1.4 g, 10.85 mmol, 1.89 mL, 3.0 equiv) was added at room temperature. The reaction mixture was refluxed for 16 h, cooled to room temperature and evaporated under reduced pressure. The residue was poured in ice and diluted with ice-cold ammonia (20 mL, 20-25% of ammonia), the product was extracted with CHCl.sub.3 (2×50 mL) and evaporated. 4-Chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 8C (1.0 g, 88.4% purity, 3.0 mmol, 82.9% yield) was obtained as yellow solid.

    Step C: rac-N-((1R,3S)-3-Methoxycyclopentyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (8)

    [0686] To the solution of 4-chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 8C (801.08 mg, 2.72 mmol) in DMSO (5 mL), N,N-diisopropylethylamine (1.05 g, 8.15 mmol, 1.42 mL, 3.0 equiv) and rac-(1R,3S)-3-methoxycyclopentan-1-amine (469.52 mg, 4.08 mmol) were added in one portion. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. The Example compound, rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (8) (50.0 mg, 96.0% purity, 128.52 μmol, 4.7% yield) was obtained as yellow viscous residue.

    Example 9

    [0687] ##STR00148##

    N-((1s,3s)-3-Methoxycyclobutyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (9)

    [0688] The 4-chloro-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 8C (801.18 mg, 2.72 mmol) was dissolved in DMSO (5 mL) and N,N-diisopropylethylamine (1.05 g, 8.15 mmol, 1.42 mL, 3.0 equiv) with corresponding amine cis-3-methoxycyclobutan-1-amine hydrochloride (561.04 mg, 4.08 mmol) was added. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. The Example compound, (9) N-((1s,3s)-3-methoxycyclobutyl)-2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (60.0 mg, 166.92 μmol, 6.1% yield) was obtained as yellow viscous residue.

    Example 10

    [0689] ##STR00149##

    Rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (10)

    Step A: 2-(1-Methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 10A

    [0690] Methyl 2-aminothiophene-3-carboxylate 8A (1.0 g, 6.36 mmol), 1-methyl-1H-imidazole-2-carbonitrile X1 (1.02 g, 9.55 mmol), potassium t-butoxide (2.14 g, 19.09 mmol) was dissolved in dry MeOH (10 mL) and stirred overnight at room temperature. The reaction mixture was evaporated and acetic acid was added to pH 5. The resulted mixture was extracted with EtOAc (25 mL), organic layer was separated, dried over Na.sub.2SO.sub.4 and evaporated to give 2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 10A (1.5 g, 92.0% purity, 5.94 mmol, 93.4% yield) which was used without additional purification on a further step.

    Step B: 4-Chloro-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 10B

    [0691] To the suspension of 2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 10A (1.75 g, 7.54 mmol) in phosphoroyl trichloride (11.56 g, 75.39 mmol, 7.03 mL, 10.0 equiv) N,N-diisopropylethylamine (1.4 g, 10.85 mmol, 1.89 mL, 3.0 equiv) was added at room temperature. The reaction mixture was refluxed for 16 h, cooled to room temperature. and evaporated under reduced pressure. The residue was poured in ice and diluted with ice-cold ammonia (20 mL, 20-25% of ammonia), the product was extracted with CHCl.sub.3 (2×50 mL) and evaporated. 4-Chloro-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 10B (1.85 g, 86.7% purity, 6.4 mmol, 84.9% yield) was obtained as yellow solid.

    Step C: rac-N-((1R,3S)-3-Methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (10)

    [0692] To the solution of 4-chloro-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 10B (1.3 g, 5.19 mmol) in DMSO (5 mL) N,N-diisopropylethylamine (1.01 g, 7.78 mmol, 1.35 mL, 1.5 equiv) and rac-(1R,3S)-3-methoxycyclopentan-1-amine (895.74 mg, 7.78 mmol) were added in one portion. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. The Example compound, rac-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (10) (90.0 mg, 273.21 μmol, 5.3% yield) was obtained as yellow viscous residue.

    Example 11

    [0693] ##STR00150##

    [0694] Rac-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (11).

    Step A: Ethyl 4-hydroxythieno[2,3-d]pyrimidine-2-carboxylate 11A

    [0695] Methyl 2-aminothiophene-3-carboxylate 8A (24.99 g, 159.01 mmol) and ethyl cyanoformate (23.63 g, 238.52 mmol, 23.4 mL, 1.5 equiv) were dissolved in dioxane*HCl (8 M solution, 350 mL) and resulted mixture was refluxed overnight. Then dioxane*HCl 250 mL (8 M solution) was added to the reaction mixture and refluxed 8 h. The reaction mixture was filtered and the product washed with CH.sub.3CN (350 mL) and MTBE (500 mL) to give ethyl 4-hydroxythieno[2,3-d]pyrimidine-2-carboxylate 11A (25.0 g, 82.0% purity, 91.42 mmol, 57.5% yield) as brown solid.

    Step B: Ethyl 4-chlorothieno[2,3-d]pyrimidine-2-carboxylate 11B

    [0696] To the suspension of ethyl 4-hydroxythieno[2,3-d]pyrimidine-2-carboxylate 11A (11.5 g, 51.29 mmol) in phosphoroyl trichloride (62.91 g, 410.28 mmol, 38.24 mL, 8.0 equiv) N,N-diispropylethylamine (19.88 g, 153.86 mmol, 26.8 mL, 3.0 equiv) was added at room temperature. The reaction mixture was refluxed for 16 h, cooled to room temperature and evaporated under reduced pressure. The residue was poured in ice and diluted with ice-cold ammonia (150 mL, 20-25% of ammonia) to pH >7.5, the product was extracted with CHCl.sub.3 (2×250 mL). Organic phase was dried over Na.sub.2SO.sub.4 and evaporated. Ethyl 4-chlorothieno[2,3-d]pyrimidine-2-carboxylate 11B (12.0 g, 90.0% purity, 44.5 mmol, 86.8% yield) was obtained as brown solid.

    Step C: rac-Ethyl 4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 11C

    [0697] To a solution of 4-chlorothieno[2,3-d]pyrimidine-2-carboxylate 11B (5.75 g, 23.69 mmol) in DMSO (5 mL), N,N-diispropylethylamine (4.59 g, 35.54 mmol, 6.19 mL, 1.5 equiv) and rac-(1R,3S)-3-methoxycyclopentan-1-amine (3.0 g, 26.06 mmol) were added in one portion. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. rac-Ethyl 4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 11C (1.5 g, 84.0% purity, 3.92 mmol, 16.5% yield) was obtained as yellow viscous residue.

    Step D: rac-4-(((1R,3S)-3-Methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (11)

    [0698] General procedure: rac-Ethyl 4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 11C (1 eq) (0.2 g, 0.622 mmol) was suspended in the appropriate corresponding amine (5 eq), (and stirred 24-72 h at room temperature. The reaction mixture was concentrated and purified by HPLC. Using NH.sub.4OH 25% as the amine produced the Example compound (11), rac-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (0.050 g, 29.9% yield).

    Example 12

    [0699] ##STR00151##

    Rac-(4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidin-2-yl)(morpholino)methanone (12)

    [0700] Following the general procedure described in Example 11, Step D using rac-ethyl 4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 11C (1 eq) (0.2 g, 0.622 mmol) and morpholine as the amine produced the Example compound (12), rac-(4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidin-2-yl)(morpholino)methanone (0.009 g, 4.3% yield).

    Example 13

    [0701] ##STR00152##

    Rac-N-isopropyl-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (13)

    Step A: rac-4-(((1R,3S)-3-Methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylic acid 13A

    [0702] To a solution of rac-ethyl 4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 11C (600.0 mg, 1.87 mmol) in THE (6 mL) and H.sub.2O (1 mL) LiOH×H.sub.2O (235.12 mg, 5.6 mmol) was added in one portion. The reaction mixture was stirred for 16 h at room temperature. Then it was diluted with water (5 mL), acidified by HCl (1N) to pH-2 and extracted by EtOAc (2×15 mL). The organic phase was dried over Na.sub.2SO.sub.4 and concentrated. rac-4-(((1R,3S)-3-Methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylic acid 13A (200.0 mg, 681.8 μmol, 39.8% yield) was obtained as white powder.

    Step B: rac-N-Isopropyl-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (13)

    [0703] General procedure: To a solution of rac-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylic acid 13A (0.1 g, 0.343 mmol, 1 eq) and HATU (0.156 g, 0.411 mmol, 1.2 eq) in DMF (20 mL/g), DIPEA (0.18 mL, 1 mmol, 3 eq) was added. The resulted mixture was stirred 30 min at room temperature and the appropriate corresponding amine (1.5 eq) was added in one portion. The final product was isolated by HPLC, without pre-workup. Using isopropylamine as the corresponding amine produced Example compound (13), rac-N-isopropyl-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (0.0131 g, 12.2% yield).

    Example 14-Example 16

    [0704] Following the general procedure described in Example 13, Step B, rac-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylic acid 13A (0.1 g, 0.343 mmol, 1 eq) was treated with the appropriate corresponding amine (1.5 eq) to produce the Example compounds (14)-(16) shown in Table A.

    TABLE-US-00003 TABLE A Amount, Ex. Structure Name Yield 14 [00153]embedded image rac-4-(((1R,3S)-3- Methoxycyclopentyl)amino)-N- (2-methoxyethyl)thieno[2,3- d]pyrimidine-2-carboxamide 0.0477 g, 42.6% 15 [00154]embedded image rac-4-(((1R,3S)-3- Methoxycyclopentyl)amino)-N- phenylthieno[2,3-d]pyrimidine- 2-carboxamide 0.0992 g, 84.8% 16 [00155]embedded image rac-4-(((1R,3S)-3- Methoxycyclopentyl)amino)-N- (pyridin-2-yl)thieno[2,3- d]pyrimidine-2-carboxamide 0.015 g, 12.7%

    Example 17

    [0705] ##STR00156##

    Rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (17)

    Step A: rac-Ethyl 6-bromo-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 17A

    [0706] To a solution of rac-ethyl 4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 11C (300.0 mg, 749.46 μmol) in DMF (5 mL), N-bromosuccinimide (160.07 mg, 899.36 μmol) was added in one portion. The resulted solution was stirred overnight at room temperature and then it was poured in ice water (10 mL). The product-precipitate formed was filtered and dried on air overnight. rac-Ethyl 6-bromo-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 17A (200.0 mg, 78.0% purity, 389.72 μmol, 55.3% yield) was obtained as dark brown powder.

    Step B: rac-Ethyl 6-(1-isopropyl-1H-pyrazol-3-yl)-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 17B

    [0707] rac-Ethyl 6-bromo-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 17A (200.39 mg, 500.6 μmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[2-(trimethylsilyl)ethoxy]methyl-1H-pyrazole (267.27 mg, 824.14 μmol), cesium carbonate (326.21 mg, 1.0 mmol) and PdXPhosG3 (42.37 mg, 50.06 μmol) was mixed in dioxane/H.sub.2O (4 mL)/(0.2 mL). The reaction flask was filled by argon and the reaction mixture was stirred at 80° C. overnight, then cooled, filtered through SiO.sub.2 pad (10 mL) and washed by EtOAc (2×15 mL). This solution was concentrated and used on next step without further purification. rac-Ethyl 6-(1-isopropyl-1H-pyrazol-3-yl)-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 17B (200.0 mg, 64.0% purity, 298.0 μmol, 63.1% yield) was obtained as brown oil.

    Step C: rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (17)

    [0708] rac-Ethyl 6-(1-isopropyl-1H-pyrazol-3-yl)-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 17B (200.0 mg, 465.62 μmol) was suspended in NH.sub.4OH (2 mL 25% w/w). This mixture was stirred 24 h at room temperature, then concentrated and purified by HPLC. The Example compound (17), rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxamide (28.0 mg, 95.0% purity, 66.42 μmol, 14.3% yield) was obtained as yellow solid.

    Example 18

    [0709] ##STR00157##

    4-Hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (18)

    Step A: (E)-3-Ethoxy-2-(pyridin-2-yl)acrylonitrile 18B

    [0710] A mixture of 2-(pyridin-2-yl)acetonitrile 18A (21.0 g, 177.76 mmol), triethoxymethane (39.52 g, 266.64 mmol, 44.4 mL, 1.5 equiv) and acetic anhydride (181.47 g, 1.78 mol, 168.03 mL, 10.0 equiv) was refluxed for 4 hours. Then it was evaporated and purified by flash chromatography to give (E)-3-ethoxy-2-(pyridin-2-yl)acrylonitrile 18B (27.0 g, 65.0% purity, 100.75 mmol, 56.7% yield).

    Step B: Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 18C

    [0711] To a stirred solution of methyl 2-sulfanylacetate (98.11 g, 924.27 mmol, 82.65 mL, 1.15 eq) in dry MeOH (500 mL), sodium methanolate (78.16 g, 1.45 mol) was added at 0° C. After 5 min to the resulted mixture a solution of (E)-3-ethoxy-2-(pyridin-2-yl)acrylonitrile 18B (140.01 g, 803.72 mmol) in MeOH (200 mL) was added dropwise. This mixture was heated at 65° C. overnight. Then the solvent was evaporated and distributed in water/EtOAc (700 mL/700 mL). Organic phase was separated, dried over Na.sub.2SO.sub.4 and evaporated. Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 18C (20.0 g, 85.37 mmol, 10.6% yield) was obtained by column chromatography.

    Step C: Ethyl 4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylate 18D

    [0712] Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 18C (4.5 g, 19.22 mmol) and ethyl cyanoformate (3.33 g, 33.64 mmol, 3.3 mL, 1.75 eq) were mixed in dioxane*HCl (8 M solution, 75 mL) and refluxed overnight. Then to the reaction mixture dioxane*HCl (8 M solution) 50 mL was added and refluxed for 8 hours. The resulted mixture was cooled to room temperature. The precipitate formed was filtered and washed with CH.sub.3CN (50 mL), MTBE (100 mL) and dried on open air to give ethyl 4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylate 18D (5.25 g, 83.5% purity, 14.55 mmol, 75.7% yield) as brown solid.

    Step D: 4-Hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (18)

    [0713] General procedure: Ethyl 4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylate 18D (0.3 g (995.62 μmol) was suspended in 5 mL of appropriate corresponding amine and the resulted mixture was stirred for 24 h at room temperature. After that, the reaction mixture was concentrated and purified by HPLC. Using 25% NH.sub.4OH as the amine produced the Example compound (18), 4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (0.0267 g, 9.9% yield).

    Example 19

    [0714] ##STR00158##

    (4-Hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-2-yl)(pyrrolidin-1-yl)methanone (19)

    [0715] Using the general procedure described in Example 18, Step D, ethyl 4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylate 18D (0.3 g, 995.62 μmol) and pyrrolidine produced the Example compound (19), (4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-2-yl)(pyrrolidin-1-yl)methanone (0.0131 g, 4% yield).

    Example 20

    [0716] ##STR00159##

    4-Hydroxy-N-isopropyl-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (20)

    Step A: 4-Hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylic acid 20A (Li.SUP.+ Salt)

    [0717] To a solution of ethyl 4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylate 18D (10.5 g, 34.85 mmol) in THE (100 mL) and H.sub.2O (20 mL) LiOH×H.sub.2O (3.66 g, 87.11 mmol) was added in one portion. The resulted mixture was stirred for 16 h at room temperature. The precipitate of desired product was filtered, washed by H.sub.2O (2×3 mL) and by THE (3 mL) and dried in open air. 4-Hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylic acid 20A (Li.sup.+ salt) (6.6 g, 24.15 mmol, 74.4% yield) was obtained as white powder.

    Step B: 4-Hydroxy-N-isopropyl-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (20)

    [0718] General procedure: Hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylic acid (Li.sup.+ salt) 20A (400.04 mg, 1.46 mmol) was dissolved in 6 M Dioxane*HCl (10 mL/g) and stirred for 15 min and evaporated. The resulted residue was dissolved in DMF (20 mL/g) then HATU (834.94 mg, 2.2 mmol) and 6 eq of DIPEA (8.78 mmol, 1.53 mL, 6.0 equiv) were added subsequently. This mixture was stirred for 30 min at room temperature then the appropriate corresponding amine (1.5 eq) was added and the reaction mixture was stirred 16 h at room temperature. The final product was purified by NPLC without pre-workup. Using isopropyl amine produced the Example compound (20), 4-hydroxy-N-isopropyl-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (0.0375 g, 6.50 yield).

    Example 21-Example 23

    [0719] Following the general procedure described in Example 20, Step B, hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylic acid (Li.sup.+ salt) 20A (400.04 mg, 1.46 mmol) and the appropriate corresponding amines (1.5 eq) produced the Example compounds (21)-(23) shown in Table B.

    TABLE-US-00004 TABLE B Amount, Ex. Structure Name Yield 21 [00160]embedded image 4-Hydroxy-N-(2- methoxyethyl)-7-(pyridin-2- yl)thieno[3,2-d]pyrimidine- 2-carboxamide 0.0575 g, 15.8% 22 [00161]embedded image (4-Hydroxy-7-(pyridin-2- yl)thieno[3,2-d]pyrimidin-2- yl)(morpholino)methanone 0.0456 g, 9.2% 23 [00162]embedded image 4-Hydroxy-N-phenyl-7- (pyridin-2-yl)thieno[3,2- d]pyrimidine-2- carboxamide 0.0113 g, 2.2%

    Example 24

    [0720] ##STR00163##

    4-Hydroxy-N,7-di(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (24)

    [0721] To a solution of 4-hydroxy-7-(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxylic acid (Li.sup.+ salt) 20A (297.23 mg, 1.09 mmol) and TCFH (457.77 mg, 1.63 mmol) in DMF, 1-methyl-1H-imidazole (267.8 mg, 3.26 mmol, 260.0 μL, 3.0 equiv) was added in one portion and the mixture was stirred 30 min at room temperature, then to the stirred mixture pyridin-2-amine (204.73 mg, 2.18 mmol) was added. After that, the reaction mixture was stirred for 16 h at room temperature and the final product was purified by HPLC without pre-workup. The Example compound (24), 4-hydroxy-N,7-di(pyridin-2-yl)thieno[3,2-d]pyrimidine-2-carboxamide (6.4 mg, 18.32 μmol, 1.7% yield) was obtained.

    Example 25

    [0722] ##STR00164##

    Rac-(4-(((1R,3S)-3-Methoxycyclopentyl)amino)thieno[2,3-d]pyrimidin-2-yl)(pyrrolidin-1-yl)methanone (25)

    [0723] Following the general procedure described in Example 11, Step D using rac-ethyl 4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidine-2-carboxylate 11C (1 eq) (0.2 g, 0.622 mmol) and pyrrolidine as the amine produced the Example compound (25), rac-(4-(((1R,3S)-3-methoxycyclopentyl)amino)thieno[2,3-d]pyrimidin-2-yl)(pyrrolidin-1-yl)methanone (0.0297 g, 14.8% yield).

    Example 26

    [0724] ##STR00165##

    6-Methyl-2,7-di(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (26)

    Step A: 7-Iodo-6-methyl-2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol 26B

    [0725] A solution of 6-methyl-2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol 26A (500.17 mg, 2.06 mmol) (for preparation see Hadari, Y. et al., WO 2018237084, Scheme 10) and N-bromosuccinimide (384.21 mg, 2.16 mmol) in acetonitrile (10 mL) was heated at reflux for 12 h. The solvent was evaporated under vacuum. The residue was diluted with water and extracted with EtOAc (6 mL). The organic extract was dried over Na.sub.2SO.sub.4, and evaporated in vacuo to afford the 7-iodo-6-methyl-2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol 26B (400.0 mg, 90.0% purity, 975.13 μmol, 47.4% yield).

    Step B: 6-Methyl-2,7-di(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (26)

    [0726] To a stirred solution of 7-iodo-6-methyl-2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol 26B (383.74 mg, 1.04 mmol) in toluene (6 mL) were added copper(I) iodide (237.55 mg, 1.25 mmol), 2-(tributylstannyl)pyridine (420.69 mg, 1.14 mmol, 370.0 μL, 1.1 equiv) under Argon atmosphere and tetrakis(triphenylphosphine)palladium(0) (60.27 mg, 51.97 μmol) was added under inert atmosphere. The mixture was heated at 100° C. for 16 h, concentrated under reduced pressure and purified by HPLC to afford the Example compound (26), 6-methyl-2,7-di(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (8.8 mg, 27.47 μmol, 2.6% yield).

    Example 27

    [0727] ##STR00166##

    7-(3-Fluoropyridin-2-yl)-2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (27)

    Step A: 2-(Pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol 27B

    [0728] Methyl 3-aminothiophene-2-carboxylate (10.0 g, 63.62 mmol) was dissolved in) Dioxane*HCl (500 mL) and pyridine-2-carbonitrile (9.93 g, 95.43 mmol) was added. The resulting mixture was stirred under reflux for 12 h. Then, the mixture was evaporated under reduced pressure. The residue was diluted with acetonitrile (100 mL) and filtered. The obtained solid was dried at 50° C. to afford 2-(Pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol 27B (9.0 g, 39.26 mmol, 61.7% yield).

    Step B: 7-Bromo-4-chloro-2-(pyridin-2-yl)thieno[3,2-d]pyrimidine 27C

    [0729] To a solution 2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol 27B (9 g, 0.0393 mol) in 90 mL of acetic acid, 10.0 mL of bromine was added at room temperature. The reaction mixture was stirred under reflux for 48 h. Then, the resulting mixture was allowed to cool to room temperature and the volatiles were evaporated under reduced pressure. The residue was diluted with water (100 mL). The resulting precipitate was filtered, washed with water (100 mL*2) and dried to give 7-bromo-4-chloro-2-(pyridin-2-yl)thieno[3,2-d]pyrimidine 27C (7.3 g, mol, 65% yield).

    Step C: 7-(3-Fluoropyridin-2-yl)-2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (27)

    [0730] 7-Bromo-4-chloro-2-(pyridin-2-yl)thieno[3,2-d]pyrimidine 27C (1.2 g, 3.89 mmol), 3-fluoro-2-(tributylstannyl)pyridine (1.8 g, 4.67 mmol), cesium fluoride (887.31 mg, 5.84 mmol) and copper(I) iodide (74.16 mg, 389.42 μmol) were mixed together in DMF (10 mL). The resulting mixture was degassed and Pd(PPh.sub.3).sub.4 (225.78 mg, 194.71 μmol) was added. The reaction mixture was heated at 100° C. overnight. The mixture was extracted with ethyl acetate (25 mL)/Brine (15 mL). The organic phase was washed with water (20 mL), dried over MgSO.sub.4 and concentrated in vacuo. The crude material was purified via HPLC (2-10 min; 55-65% water+FA-acetonitrile+FA; Flow rate 30 mL/min; loading pump 4 mL/min acetonitrile; column SunFire 19*100 mm) to provide 7-(3-fluoropyridin-2-yl)-2-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol.

    Examples 28-29

    [0731] The following compounds are offered to illustrate but not to limit the disclosure, and may be for use in the compositions and methods provided herein:

    ##STR00167##

    Example 30

    [0732] ##STR00168##

    7-(3-Fluoropyridin-2-yl)-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidin-4-ol (30)

    Step A: 7-Bromo-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidin-4-ol 30B

    [0733] Sodium hydride (201.6 mg, 8.4 mmol) was dissolved in DMF (5 mL) and methyl 3-amino-4-bromothiophene-2-carboxylate 30A (991.67 mg, 4.2 mmol) was added. The resulting mixture was stirred at room temperature for 2 h following by the addition of 1-methyl-1H-imidazole-2-carbonitrile (450.0 mg, 4.2 mmol, 450.0 μl, 1.0 equiv). The reaction mixture was heated at 80° C. for 16 h. Then, the mixture was evaporated under reduced pressure, diluted with water (10 mL) and extracted with EtOAc (15 mL*3). The organic layer dried over Na.sub.2SO.sub.4 and evaporated to give 7-bromo-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidin-4-ol 30B (300.0 mg, 964.14 μmol, 23% yield).

    Step B: 7-(3-Fluoropyridin-2-yl)-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidin-4-ol (30)

    [0734] 7-Bromo-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidin-4-ol 30B (350.0 mg, 1.12 mmol), 2-methyl-6-(tributylstannyl)pyridine (472.61 mg, 1.24 mmol) and copper(I) iodide (21.41 mg, 112.42 μmol) were mixed together in DMF (5 mL). The mixture was degassed and Pd(PPh.sub.3).sub.4 (130.36 mg, 112.42 μmol) was added. The reaction mixture was heated at 100° C. overnight. The mixture was extracted with ethyl acetate (25 mL)/brine (15 mL). The organic phase was washed with water (20 mL), dried over MgSO.sub.4 and concentrated in vacuo. The crude material was purified via HPLC (2-10 min; 10-50% MeCN/Water+FA; Flow rate 30 mL/min (loading pump 4 mL MeCN); column: SunFire 100*19 mm, 5 microM) to provide 7-(3-fluoropyridin-2-yl)-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidin-4-ol (30) (3.0 mg, 91.4% purity, 8.71 μmol, 0.8% yield).

    Example 31

    [0735] ##STR00169##

    6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1S,3R)-3-methoxycyclopentyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (31)

    Step A: Methyl 2-[(tert-butoxy)carbonyl]amino-4-methylthiophene-3-carboxylate 31B

    [0736] DMAP (642.19 mg, 5.26 mmol) was added to a stirred solution of methyl 2-amino-4-methylthiophene-3-carboxylate 31A (9.0 g, 52.56 mmol) and Boc.sub.2O (13.77 g, 63.08 mmol) in 108 mL of dioxane. The mixture was refluxed overnight. After 16 h reaction was complete (monitored by NMR). The mixture was concentrated under reduced pressure and purified by column chromatography (Eluted by Hex:EtOAc 10:1). Methyl 2-[(tert-butoxy)carbonyl]amino-4-methylthiophene-3-carboxylate 31B (8.1 g, 29.85 mmol, 56.8% yield) was obtained as white solid.

    Step B: Methyl 5-bromo-2-[(tert-butoxy)carbonyl]amino-4-methylthiophene-3-carboxylate 31C

    [0737] To a solution of methyl 2-[(tert-butoxy)carbonyl]amino-4-methylthiophene-3-carboxylate 31B (8.1 g, 29.85 mmol) in 160 mL of DMF, N-bromosuccinimide (6.38 g, 35.82 mmol) was added in one portion. The resulting mixture was stirred at room temperature overnight. Then, the mixture was allowed to cool to room temperature and poured in ice water (350 mL). The obtained precipitate was filtered, washed 3 times with water and dried on air. Methyl 5-bromo-2-[(tert-butoxy)carbonyl]amino-4-methylthiophene-3-carboxylate 31C (9.2 g, 26.27 mmol, 88% yield) was obtained as light-yellow powder.

    Step C: Methyl 2-((tert-butoxycarbonyl)amino)-5-(1-isopropyl-1H-pyrazol-3-yl)-4-methylthiophene-3-carboxylate 31D

    [0738] Methyl 5-bromo-2-[(tert-butoxy)carbonyl]amino-4-methylthiophene-3-carboxylate 31C (2.0 g, 5.71 mmol), 1-(propan-2-yl)-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.62 g, 6.85 mmol), cesium carbonate (3.72 g, 11.42 mmol) and (2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (966.74 mg, 1.14 mmol) were mixed together in degassed dioxane:H.sub.2O (40 mL:2 mL) and refluxed overnight. Then, the mixture was allowed to cool to room temperature, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO.sub.2; Elute by Hexane:EtOAc 10:1 to 1:10). Methyl 2-((tert-butoxycarbonyl)amino)-5-(1-isopropyl-1H-pyrazol-3-yl)-4-methylthiophene-3-carboxylate 31D (1.2 g, 90.0% purity, 2.85 mmol, 49.8% yield) was obtained as orange powder.

    Step D: Methyl 2-amino-5-(1-isopropyl-1H-pyrazol-3-yl)-4-methylthiophene-3-carboxylate 31E

    [0739] Methyl 2-((tert-butoxycarbonyl)amino)-5-(1-isopropyl-1H-pyrazol-3-yl)-4-methylthiophene-3-carboxylate 31D (1.2 g, 3.16 mmol) was dissolved in 5 mL of dioxane and 6M dioxane*HCl was added dropwise at 0° C. The reaction mixture was stirred overnight at room temperature. Then, the volatiles were removed in vacuo and the residue was purified by flash chromatography (SiO.sub.2; Eluted by EtOAc). Methyl 2-amino-5-(1-isopropyl-1H-pyrazol-3-yl)-4-methylthiophene-3-carboxylate 31E (530.0 mg, 90.0% purity, 1.71 mmol, 57.3% yield) was obtained as dark-grey gum.

    Step E: 6-(1-Isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 31F

    [0740] Methyl 2-amino-5-(1-isopropyl-1H-pyrazol-3-yl)-4-methylthiophene-3-carboxylate 31E (430.35 mg, 1.54 mmol), 1-methyl-1H-imidazole-2-carbonitrile (247.51 mg, 2.31 mmol) and potassium 2-methylpropan-2-olate (691.45 mg, 6.16 mmol) were dissolved in dry MeOH (5 mL). The reaction mixture was stirred overnight at room temperature. The residue was evaporated under reduced pressure and HOAc (conc) was added to pH 5. Then, the solution was mixed with EtOAc (15 mL) and extracted. The organic phase was dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure to afford 6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 31F (400.0 mg, 80.0% purity, 902.86 μmol, 58.6% yield).

    Step F: 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31G

    [0741] The crude material from the previous step (6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-ol 31F) (100.0 mg, 282.14 μmol)) was suspended in POCl.sub.3 (2 mL) and diisopropylethylamine (0.6 mL) was added at room temperature. The reaction mixture was stirred under reflux for 16 h. The mixture was allowed to cool to the room temperature and evaporated under reduced pressure. The residue was poured into ice and basified with liquid ammonia (20 mL, 20-25% of ammonia) and filtered on to afford 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31G (100.0 mg, 55.0% purity, 147.5 μmol, 52.4% yield) as brown liquid.

    Step G: General Procedure: 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31G

    [0742] (100.0 mg, 268.19 μmol, 1.0 equiv), corresponding amine (1.5 equiv) and ethylbis(propan-2-yl)amine (4 equiv) were mixed in DMSO (2 mL). The resulting mixture was stirred at 100° C. for 12 hours and then allowed to cool to room temperature. The mixture was diluted with water (10 mL) and extracted several times with EtOAc (3*15 mL). The combined extracts were washed with brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was evaporated. The residue was subjected for HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-8 min 50-75% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)).

    Step H: 6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1S,3R)-3-methoxycyclopentyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (31)

    [0743] 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31G (1.88 g, 4.33 mmol) was dissolved in DMSO (18 mL) and ethylbis(propan-2-yl)amine (2.24 g, 17.34 mmol, 3.02 ml, 4.0 equiv) was added. The resulting mixture was stirred for 5 min at room temperature following by the addition of (1S,3R)-3-methoxycyclopentan-1-amine 4F (prepared as described in Example 4, Step C) (499.12 mg, 4.33 mmol). The reaction mixture was stirred at 100° C. overnight. Then, the mixture was allowed to cool to the room temperature and subjected to HPLC (2-10 min; 65-80% water-MeOH; Flow rate 30 mL/min; loading pump 4 mL/min methanol; column: SunFire 100*19 mm, 5 microM) purification to afford 6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1S,3R)-3-methoxycyclopentyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (31). (390.0 mg, 95.0% purity, 721.3 μmol, 16.6% yield) as yellow gum.

    Example 32

    [0744] ##STR00170##

    6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (32)

    [0745] 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31G (428.54 mg, 1.15 mmol) was dissolved in DMSO (18 mL) and ethylbis(propan-2-yl)amine (593.6 mg, 4.59 mmol, 800.0 μl, 4.0 equiv) was added. The resulting mixture was stirred for 5 min at room temperature following by the addition of (1R,3S)-3-methoxycyclopentan-1-amine 4G (prepared as described in Example 4, Step C) (145.6 mg, 1.26 mmol). The reaction mixture was stirred at 100° C. overnight. Then, the mixture was allowed to cool to the room temperature and subjected to HPLC (2-8 min; 0-75% water-MeOH; Flow rate 30 mL/min; loading pump 4 mL/min MeOH; column SunFire 19*100 mm) to afford 6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (32) (83.0 mg, 95.0% purity, 174.61 μmol, 15.2% yield) as yellow gum.

    Examples 33-63

    [0746] Following the general procedure described in Example 31, Step G, 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31G (100.0 mg, 268.19 μmol, 1.0 equiv) was treated with the appropriate corresponding amine (1.5 eq) to produce the Example compounds (33)-(63) shown in Table C.

    TABLE-US-00005 TABLE C Amount Example Structure Name (Yield) 33 [00171]embedded image rac-6-(1-Isopropyl- 1H-pyrazol-3-yl)-N- ((1R,3R)-3- methoxycyclopentyl)- 5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidin-4-amine 62.0 mg, (16.2%) 34 [00172]embedded image rac-6-(1-Isopropyl- 1H-pyrazol-3-yl)-4- (3- methoxypyrrolidin-1- yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidine 58.0 mg (15.7%) 35 [00173]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N-(2- methoxyethyl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 160.0 mg (17.8%) 36 [00174]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N-(3- methoxypropyl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 150.0 mg (14.5%) 37 [00175]embedded image rac-6-(1-Isopropyl- 1H-pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2-yl)-N- (tetrahydrofuran-3- yl)thieno[2,3- d]pyrimidin-4-amine 62.7 mg (18.4%) 38 [00176]embedded image rac-6-(1-Isopropyl- 1H-pyrazol-3-yl)-4- (3- (methoxymethyl) pyrrolidin-1-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidine 65.3 mg (17.1%) 39 [00177]embedded image rac-4-((3S,4R)-3,4- Dimethoxypyrrolidin- 1-yl)-6-(1-isopropyl- 1H-pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidine 14.6 mg (11.7%) 40 [00178]embedded image rac-6-(1-Isopropyl- 1H-pyrazol-3-yl)-4- (3-methoxypiperidin- 1-yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidine 19.6 mg (14.7%) 41 [00179]embedded image rac-4-(6-(1- Isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-yl)-3- (methoxymethyl) morpholine 15.8 mg (12.8%) 42 [00180]embedded image N-((1r,3R,4S)-3,4- Dimethoxycyclopentyl)- 6-(1-isopropyl- 1H-pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 19.4 mg (16.3%) 43 [00181]embedded image N-((1s,3R,4S)-3,4- Dimethoxycyclopentyl)- 6-(1-isopropyl- 1H-pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 9.0 mg (8.1%) 44 [00182]embedded image rac-N-((3S,4S)-3,4- Dimethoxycyclopentyl)- 6-(1-isopropyl- 1H-pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 15.4 mg (12.2%) 45 [00183]embedded image rac-4-(6-(1- Isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-yl)-2- (methoxymethyl) morpholine 36.2 mg (28.8%) 46 [00184]embedded image N-(2-(1H-Tetrazol-5- yl)ethyl)-6-(1- isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 10.3 mg (8.2%) 47 [00185]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N- (((1s,3s)-3- methoxycyclobutyl) methyl)-5-methyl-2- (1-methyl-1H- imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 21.7 mg (16.6%) 48 [00186]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N- (((1r,3r)-3- methoxycyclobutyl) methyl)-5-methyl-2- (1-methyl-1H- imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 24.8 mg (19.8%) 49 [00187]embedded image (S)-6-(1-Isopropyl- 1H-pyrazol-3-yl)-4- (3-methoxypiperidin- 1-yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidine 16.3 mg (13.8%) 50 [00188]embedded image (R)-6-(1-Isopropyl- 1H-pyrazol-3-yl)-4- (3-methoxypiperidin- 1-yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidine 18.6 mg (14.3%) 51 [00189]embedded image (S)-5-((6-(1- Isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4- yl)amino)methyl) pyrrolidin-2-one 18.0 mg (14.4%) 52 [00190]embedded image (R)-5-(((6-(1- Isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4- yl)amino)methyl) pyrrolidin-2-one 14.4 mg (11.2%) 53 [00191]embedded image N-(2-(1H-Imidazol-2- yl)ethyl)-6-(1- isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 11.9 mg (9.5%) 54 [00192]embedded image 6-(1-isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2-yl)-4- (((1r,4r)-4-(oxo-λ.sup.6- methyl)cyclohexyl) amino)thieno[2,3- d]pyrimidine 13.4 mg (10.2%) 55 [00193]embedded image 6-(1-isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2-yl)-4- (((1s,4s)-4-(oxo-λ.sup.6- methyl)cyclohexyl) amino)thieno[2,3- d]pyrimidine 7.7 mg (9.2%) 56 [00194]embedded image rac-5-(2-((6-(1- Isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4- yl)amino)ethyl) pyrrolidin-2-one 5.7 mg (4.6%) 57 [00195]embedded image N-((1H-Imidazol-2- yl)methyl)-6-(1- isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 17.4 mg (13.8%) 58 [00196]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2-yl)-N- (2-(1-methyl-1H- imidazol-4- yl)ethyl)thieno[2,3- d]pyrimidin-4-amine 18.7 mg (14.5%) 59 [00197]embedded image N-((1H-Imidazol-4- yl)methyl)-6-(1- isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 3.3 mg (2.6%) 60 [00198]embedded image 5-(((6-(1-Isopropyl- 1H-pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4- yl)amino)methyl)-4- methyl-2,4-dihydro- 3H-1,2,4-triazol-3- one 11.2 mg (9.0%) 61 [00199]embedded image rac-6-(1-Isopropyl- 1H-pyrazol-3-yl)-N- (((1S,2R)-2- methoxycyclobutyl) methyl)-5-methyl-2- (1-methyl-1H- imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 5.3 mg (4.2%) 62 [00200]embedded image rac-4-(3- (Isopropoxymethyl) pyrrolidin-1-yl)-6-(1- isopropyl-1H- pyrazol-3-yl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidine 9.3 mg (7.2%) 63 [00201]embedded image rac-6-(1-Isopropyl- 1H-pyrazol-3-y)-N- (((1S,2R)-2- (methoxymethyl)cyclo- butyl)methyl)-5- methyl-2-(1-methyl- 1H-imidazol-2- yl)thieno[2,3- d]pyrimidin-4-amine 20.1 mg (7.5%)

    Example 64

    [0747] ##STR00202##

    6-(1-Isopropyl-1H-pyrazol-3-yl)-N-(3-methoxyphenyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (64)

    [0748] General Procedure: To a solution of 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31F (prepared as described in Example 31, Step E) (100.0 mg, 268.19 μmol, 1 equiv) and the appropriate corresponding amine (1.5 equiv) in dry DMF (1 mL) sodium hydride (13.42 mg, 559.11 μmol, 1.5 eq) was added under an Ar atmosphere at 0° C. The reaction mixture was stirred at 50° C. for 18 h. Then, the mixture was allowed to cool to room temperature, diluted with water (2 mL) and concentrated to dryness under reduced pressure. The residue was purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-8 min 50-75% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford pure product. Using 3-methoxyaniline as the corresponding amine produced 6-(1-isopropyl-1H-pyrazol-3-yl)-N-(3-methoxyphenyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (64) (17.2 mg, 16.7% yield).

    Examples 65-68

    [0749] Following the general procedure described in Example 65, Step A, 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-pH-imidazol-2-yl)thieno[2,3-d]pyrimidine 31F (100.0 mg, 268.19 μmol, 1.0 equiv) (100.0 mg, 268.19 μmol, 1 equiv) and the appropriate corresponding amine (1.5 equiv), produced the Example compounds (65)-(68) shown in Table D.

    TABLE-US-00006 TABLE D Amount Example Structure Name (Yield) 65 [00203]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N-(6- methoxypyridin-2- yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidin-4-amine hydrochloride salt 6.6 mg (6.4%) 66 [00204]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N-(6- methoxypyrimidin-4- yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidin-4-amine 29.0 mg (7.8%) 67 [00205]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N-(2- methoxypyridin-4- yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidin-4-amine 30.5 mg (23.1%) 68 [00206]embedded image 6-(1-Isopropyl-1H- pyrazol-3-yl)-N-(4- methoxypyrimidin-2- yl)-5-methyl-2-(1- methyl-1H-imidazol- 2-yl)thieno[2,3- d]pyrimidin-4-amine 32.6 mg (15.3%)

    Example 69

    [0750] ##STR00207##

    Rac-6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-5-methyl-2-(pyridine-2-yl)thieno[2,3-d]pyrimidin-4-amine (69)

    Step A: 6-(1-Isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidin-4-ol 69A

    [0751] Pyridine-2-carbonitrile (532.8 mg, 5.12 mmol) was dissolved in DMF (10 mL) and sodium hydride (558.24 mg, 23.26 mmol) was added. The resulting mixture was stirred at room temperature for 2 h following by the addition methyl 2-amino-5-(1-isopropyl-1H-pyrazol-3-yl)-4-methylthiophene-3-carboxylate 31E (prepared as described in Example 31, Step D) (1.3 g, 4.65 mmol). The reaction mixture was stirred at 80° C. for 16 h. Then the volatiles were removed in vacuo. The residue was diluted with water (10.0 mL) and extracted with EtOAc (15.0 mL*3). The organic layer was concentrated under reduced pressure to afford 6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidin-4-ol 69A (1.0 g, 2.85 mmol, 61.2% yield).

    Step B: 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine 69B

    [0752] 6-(1-Isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidin-4-ol 69A (1.0 g, 2.85 mmol) was suspended in phosphoroyl trichloride (3.5 g, 22.82 mmol, 2.13 mL, 8.0 equiv) and ethylbis(propan-2-yl)amine (1.11 g, 8.55 mmol, 1.49 mL, 3.0 equiv) was added at room temperature. The reaction mixture was stirred under reflux for 16 h. Then, the resulting mixture was allowed to cool to the room temperature and concentrated under reduced pressure. The residue was poured in ice and diluted with ice-cold ammonia (20 mL, 20-25% of ammonia). The product was extracted with chloroform (50.0 mL*2) and evaporated in vacuo to afford 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine 69B (800.0 mg, 2.16 mmol, 75.8% yield) as brown slurry.

    Step C: rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidin-4-amine (69)

    [0753] rac trans-3-Methoxycyclopentan-1-amine (60.64 mg, 526.51 μmol), ethylbis(propan-2-yl)amine (185.5 mg, 1.44 mmol, 250.0 μl, 3.0 equiv) and 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine 69B (178.0 mg, 478.65 μmol) were mixed in DMSO (2.0 mL). The resulting reaction mixture was stirred at 100° C. for 15 hours and then allowed to cool to room temperature. The mixture was diluted with water (10 mL) and extracted three times with EtOAc (15 mL). The combined extracts were washed with brine (10 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min, 0-60% MeOH—H.sub.2O flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford rac-6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidin-4-amine (69) (10.0 mg, 95.0% purity, 21.18 μmol, 4.6% yield).

    Example 70

    [0754] ##STR00208##

    Rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-4-(3-methoxypiperidin-1-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine (70)

    [0755] 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine 69B (prepared as described in Example 69, Step C) (249.83 mg, 675.46 μmol), rac 3-methoxypiperidine hydrochloride (153.63 mg, 1.01 mmol) and ethylbis(propan-2-yl)amine (348.74 mg, 2.7 mmol, 470.0 μL, 4.0 equiv) were mixed in DMSO (5 mL). The resulting reaction mixture is stirred at 100° C. for 12 hours and then allowed to cool to room temperature. The mixture is diluted with water (20 ml) and extracted several times with EtOAc (3*25 mL). The combined extracts were washed with brine (25 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was subjected for HPLC purification (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-85% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-methoxypiperidin-1-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine (70) (85.0 mg, 189.49 μmol, 28.1% yield).

    Example 71

    [0756] ##STR00209##

    Rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-4-(3-(methoxymethyl)pyrrolidin-1-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine (71)

    [0757] 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine 69B (prepared as described in Example 69, Step B) (148.42 mg, 401.26 μmol), rac 3-(methoxymethyl)pyrrolidine (69.32 mg, 601.89 μmol) and ethylbis(propan-2-yl)amine (155.82 mg, 1.21 mmol, 210.0 μL, 3.0 equiv) were mixed in DMSO (5 mL). The resulting reaction mixture is stirred at 100° C. for 12 hours and then allowed to cool to room temperature. The mixture is diluted with water (20 mL) and extracted several times with EtOAc (3*25 mL). The combined extracts were washed with brine (25 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was subjected for HPLC purification (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-80% MeCN-water; flow 30 mL/min (loading pump 4 mL/min MeCN)) to afford rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-(methoxymethyl)pyrrolidin-1-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine (71) (59.0 mg, 95.0% purity, 124.95 μmol, 31.1% yield).

    Example 72

    [0758] ##STR00210##

    4-(3-(Isopropoxymethyl)pyrrolidin-1-yl)-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine (72)

    [0759] 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine 69B (prepared as described in Example 69, Step B) (175.38 mg, 474.16 μmol), 3-[(propan-2-yloxy)methyl]pyrrolidine (101.87 mg, 711.23 μmol) and ethylbis(propan-2-yl)amine (244.86 mg, 1.89 mmol, 330.0 μL, 4.0 equiv) were mixed in DMSO (5 mL). The resulting reaction mixture is stirred at 100° C. for 12 hours and then allowed to cool to room temperature. The mixture was diluted with water (20 mL) and extracted several times with EtOAc (3*25 mL). The combined extracts were washed with brine (25 mL), dried over anhydrous sodium sulfate, and filtered. The solvent was removed in vacuo. The residue was subjected for HPLC purification (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 35-65% MeCN-water+TFA; flow 30 mL/min (loading pump 4 mL/min MeCN)) to afford 4-(3-(isopropoxymethyl)pyrrolidin-1-yl)-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidine (72) (7.6 mg, 95.0% purity, 15.15 μmol, 3.2% yield).

    Example 73

    [0760] ##STR00211##

    Rac-6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidin-4-amine (73)

    Step A: Ethyl 2-amino-5-bromo-4-(trifluoromethyl)thiophene-3-carboxylate 73B

    [0761] N-Bromosuccinimide (4.46 g, 25.08 mmol) was added portionwise to the solution of ethyl 2-amino-4-(trifluoromethyl)thiophene-3-carboxylate 73A (5.0 g, 20.9 mmol) in DMF (100 mL) at 0° C. The reaction mixture was stirred at room temperature for 16 h, then poured in ice water. The formed precipitate was collected by filtration, washed by water (2*100 mL) and dried in vacuo. Ethyl 2-amino-5-bromo-4-(trifluoromethyl)thiophene-3-carboxylate 73B (6.1 g, 90.0% purity, 17.26 mmol, 82.6% yield) was obtained.

    Step B: Ethyl 2-amino-5-(1-isopropyl-1H-pyrazol-3-yl)-4-(trifluoromethyl)thiophene-3-carboxylate 73C

    [0762] Ethyl 2-amino-5-bromo-4-(trifluoromethyl)thiophene-3-carboxylate 73B (6.1 g, 19.18 mmol), 1-(propan-2-yl)-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (5.43 g, 23.01 mmol), cesium carbonate (12.5 g, 38.35 mmol) and Pd(dppf)Cl.sub.2*CH.sub.2Cl.sub.2 (1.57 g, 1.92 mmol) were dissolved in degassed dioxane (80 mL) under Ar atmosphere. 3.8 mL of water was added via syringe. The reaction mixture was heated to 80° C. and stirred at this temperature for 16 h. The mixture was cooled to room temperature, filtered through celite and concentrated under reduced pressure. The residue was purified by column chromatography (SiO.sub.2; Eluted by Hex:EtOAc 20:1 to 1:2). Ethyl 2-amino-5-(1-isopropyl-1H-pyrazol-3-yl)-4-(trifluoromethyl)thiophene-3-carboxylate 73C (2.5 g, 7.2 mmol, 37.5% yield) was obtained as light-yellow powder.

    Step C: 6-(1-Isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidin-4-ol 73D

    [0763] Sodium methanolate (1.17 g, 21.59 mmol) was added portionwise to the solution of ethyl 2-amino-5-(1-isopropyl-1H-pyrazol-3-yl)-4-(trifluoromethyl)thiophene-3-carboxylate 73C (2.5 g, 7.2 mmol) and 1-methyl-1H-imidazole-2-carbonitrile X1 (770.83 mg, 7.2 mmol, 770.0 μl, 1.0 equiv) in MeOH (40 mL). The reaction mixture was heated to reflux and stirred at this temperature for 16 h. Then, the mixture was cooled to room temperature and neutralized by acetic acid (2.16 g, 35.98 mmol). The solution was concentrated, dissolved in 50 mL EtOAc, washed by brine (2*50 mL), dried over sodium sulphate and evaporated under reduced pressure. 6-(1-Isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidin-4-ol 73D (1.2 g, 50.0% purity, 1.47 mmol) was obtained and used in next step without further purification.

    Step D: 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine 73E

    [0764] Ethyl bis(propan-2-yl)amine (949.23 mg, 7.34 mmol) was added dropwise to the solution of 6-(1-Isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidin-4-ol 73D (1.2 g, 2.94 mmol) in phosphoroyl trichloride (9.01 g, 58.76 mmol). The mixture was heated to 100° C. and stirred at this temperature for 16 h. The reaction mixture was concentrated in vacuo, poured in ice water (5 mL) and neutralized by K.sub.2CO.sub.3. The formed precipitate was collected by filtration, washed by water (2*5 mL) and dried in vacuo. 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine 73E (850.0 mg, 30.0% purity, 597.41 μmol) was obtained. Product was used in next steps without further purification. Step E: rac-6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidin-4-amine (73). Ethylbis(propan-2-yl)amine (133.96 mg, 1.04 mmol) was added in one portion to the solution of 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine 73E (370.0 mg, 866.82 μmol) and rac-(1S,3R)-3-methoxycyclopentan-1-amine hydrochloride (47.15 mg, 310.94 μmol) in DMF (1 mL). The mixture was heated to 100° C. and stirred at this temperature for 16 h. The mixture was purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-85% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford rac-6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1R,3S)-3-methoxycyclopentyl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidin-4-amine (73). (43.0 mg, 85.05 μmol, 32.8% yield).

    Example 74

    [0765] ##STR00212##

    Rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-4-(3-(methoxymethyl)pyrrolidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine (74)

    [0766] Ethylbis(propan-2-yl)amine (245.42 mg, 1.9 mmol, 330.0 μl, 3.0 equiv) was added in one portion to the solution of 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine 73E (prepared as described in Example 73, Step D) (900.0 mg, 2.11 mmol) and rac 3-(methoxymethyl)pyrrolidine (87.48 mg, 759.55 μmol) in DMF (1 mL). The resulting mixture was heated to 100° C. and stirred at this temperature for 16 h. The mixture was purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 65-80% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-(methoxymethyl)pyrrolidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine (74) (70.0 mg, 138.46 μmol, 21.9% yield).

    Example 75

    [0767] ##STR00213##

    Rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine (75)

    [0768] Ethylbis(propan-2-yl)amine (83.54 mg, 646.41 μmol, 110.0 μl, 4.0 equiv) was added in one portion to the solution of 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine 73E (prepared as described in Example 73, Step D) (230.0 mg, 538.84 μmol) and rac 3-methoxypiperidine hydrochloride (29.41 mg, 193.92 μmol) in DMF (1 ml). The mixture was heated to 100° C. and stirred at this temperature for 16 h. The obtained mixture was purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-85% MeOH-water; flow 30 ml/min (loading pump 4 ml/min MeOH)) to afford rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-(trifluoromethyl)thieno[2,3-d]pyrimidine (75) (60.0 mg, 118.68 μmol, 73.4% yield).

    Example 76

    [0769] ##STR00214##

    Rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-(methoxymethyl)pyrrolidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine (76)

    Step A: 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 76A

    [0770] 6-Bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 1D (prepared as described in Example 1, Step C) (900.0 mg, 2.22 mmol), 3-(methoxymethyl)pyrrolidine (383.39 mg, 3.33 mmol) and ethylbis(propan-2-yl)amine (860.44 mg, 6.66 mmol, 1.16 ml, 3.0 equiv) were mixed in DMSO (5 mL). The resulting mixture was stirred at 100° C. for 12 hours and then allowed to cool to room temperature. The mixture was diluted with water (20 mL) and extracted several times with EtOAc (3*25 mL). The combined extracts were washed with brine (25 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was subjected to HPLC purification (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-85% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford 4-chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 76A (600.0 mg, 91.0% purity, 1.13 mmol, 50.8% yield).

    Step B: rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-4-(3-(methoxymethyl)pyrrolidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine (76)

    [0771] 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 76A (605.23 mg, 1.26 mmol), 1-(propan-2-yl)-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (405.51 mg, 1.72 mmol), cesium carbonate (932.61 mg, 2.86 mmol) and Pd(dppf)Cl.sub.2*CH.sub.2Cl.sub.2 (116.87 mg, 143.12 μmol) were dissolved in degassed dioxane (0.4 mL) under an Ar atmosphere. 0.018 mL of water was added via syringe. The reaction mixture was heated to 80° C. and stirred at this temperature for 16 h. The mixture was cooled to room temperature, filtered through celite and concentrated under reduced pressure. The residue was purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-85% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH) to obtain rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-(methoxymethyl)pyrrolidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine (76) (180.0 mg, 350.43 μmol, 28% yield).

    Example 77

    [0772] ##STR00215##

    Rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine (77)

    Step A: rac-6-Bromo-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 77A

    [0773] 6-Bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 1D (prepared as described in Example 1, Step C) (500.83 mg, 1.23 mmol), racemic 3-methoxypiperidine hydrochloride (280.79 mg, 1.85 mmol) and ethylbis(propan-2-yl)amine (638.12 mg, 4.94 mmol, 860.0 μL, 4.0 equiv) were mixed in DMSO (5 mL). The resulting reaction mixture is stirred at 100° C. for 12 hours and then allowed to cool to room temperature. The mixture is diluted with water (20 mL) and extracted several times with EtOAc (3*25 mL). The combined extracts were washed with brine (25 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified using HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-85% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford rac-bromo-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 77A (430.0 mg, 887.67 μmol, 71.9% yield).

    Step B: rac-6-(1-Isopropyl-1H-pyrazol-3-yl)-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine (77)

    [0774] rac-Bromo-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine 77A (430.0 mg, 1.03 mmol), 1-(propan-2-yl)-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (290.55 mg, 1.23 mmol), cesium carbonate (668.22 mg, 2.05 mmol) and Pd(dppf)Cl.sub.2*CH.sub.2Cl.sub.2 (83.74 mg, 102.54 μmol) were dissolved in degassed dioxane (0.4 mL) under an Ar atmosphere. 0.018 mL of water was added via syringe. The reaction mixture was heated to 80° C. and stirred at this temperature for 16 h. The mixture was cooled to room temperature, filtered through celite and concentrated in vacuo. The residue was purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min 70-85% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford rac-6-(1-isopropyl-1H-pyrazol-3-yl)-4-(3-methoxypiperidin-1-yl)-2-(1-methyl-1H-imidazol-2-yl)-5-phenylthieno[2,3-d]pyrimidine (77) (121.8 mg, 237.12 μmol, 26.5% yield).

    Example 78

    [0775] ##STR00216##

    2-(1-(2-Methoxyethyl)-1H-imidazol-4-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (78)

    Step A: (E)-3-Ethoxy-2-(pyridin-2-yl)acrylonitrile 78B

    [0776] 2-(Pyridin-2-yl)acetonitrile 78A (32.00 g, 1 equiv) and triethyl orthoformate (40 g, 1 equiv) was added to acetic anhydride (55.3 g, 2 equiv) at room temperature. The resulting reaction mixture was heated at 100° C. for 3 h, cooled to room temperature, diluted with water (500 mL), and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4, and chromatographed on silica gel yielding (E)-3-ethoxy-2-(pyridin-2-yl)acrylonitrile 78B as a brown solid (16.4 g, 35%).

    Step B: Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C

    [0777] Methyl mercaptoacetate (7.2 g, 1.1 equiv) was dissolved in dry THE (200 mL) and cooled to 0° C. DBU (14.1 g, 1.5 equiv) was added thereto portion wise followed by slow addition of (E)-3-ethoxy-2-(pyridin-2-yl)acrylonitrile 78B (10.7 g, 1 equiv). The mixture was stirred at room temperature overnight, the solvent was evaporated and the residue was chromatographed on silica gel yielding methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C as a brownish solid (6.2 g, 43%).

    Step C: 2-(1-(2-Methoxyethyl)-1H-imidazol-4-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (78)

    [0778] General Procedure A: Sodium hydride (2.5 eq) (60% in mineral oil) was added portionwise (during 3 min) to the solution of methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (300.0-500.0 mg) and the correct corresponding nitrile HetArCN X6 (1 eq) in dioxane (6 mL). The reaction mixture was heated to reflux and stirred at this temperature for 16 h. Then the mixture was cooled to the room temperature, poured in ice water and neutralized by acetic acid to pH 6-4. The formed participate was collected by filtration, washed by water (2*5 mL) and purified by HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-8 min 50-75% MeOH-water; flow 30 mL/min (loading pump 4 mL/min MeOH)). Products of cyclization were obtained with good yields (4-35%). Using 1-(2-methoxyethyl)-1H-imidazole-4-carbonitrile X7 as the HetArCN nitrile produced 2-(1-(2-methoxyethyl)-1H-imidazol-4-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (78) (19.1 mg, 4.3% yield). General Procedure B: Sodium hydride (60% dispertion in mineral oil; 2.0 equiv) was added portionwise (during 3 min) to the solution of methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (0.2 g, 8 mmol, 1.0 equiv) and corresponding HetArCN nitrile X7 (1.0 equiv) in dioxane (10.0 mL). The reaction mixture was heated under reflux for 16 hours. Then, the resulting mixture was allowed to cool to room temperature, poured in ice and water and neutralized with acetic acid. The formed precipitate was collected by filtration, washed with water (2*10.0 mL) and dried in vacuo. The obtained product was subjected to HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min, 0-60% MeOH—H.sub.2O flow 30 mL/min (loading pump 4 mL/min MeOH)) to afford pure product.

    Examples 79-82

    [0779] Following the procedure described in Example 78, Step C, using General Procedure A, methyl 3-amino-4-(pyri din-2-yl)thiophene-2-carboxylate 78C (300.0-500.0 mg) was treated with the appropriate corresponding corresponding nitrile HetArCN X6 (1 eq) to produce the Example compounds (79)-(82) shown in Table E.

    TABLE-US-00007 TABLE E Amount Example Structure Name (Yield) 79 [00217]embedded image 2-(1-(3- Methoxypropyl)-1H- imidazol-2-yl)-7- (pyridin-2- yl)thieno[3,2- d]pyrimidin-4-ol 117.0 mg (24.8%) 80 [00218]embedded image 2-(1-(2-Ethoxyethyl)- 1H-imidazol-2-yl)-7- (pyridin-2- yl)thieno[3,2- d]pyrimidin-4-ol 186.2 mg (27.9%) 81 [00219]embedded image 7-(Pyridin-2-yl)-2-(1- (2-(2,2,2- trifluoroethoxy)ethyl)- 1H-imidazol-2- yl)thieno[3,2- d]pyrimidin-4-ol 123.5 mg (21.4%) 82 [00220]embedded image 2-(1-(2- Isopropoxyethyl)-1H- imidazol-2-yl)-7- (pyridin-2- yl)thieno[3,2- d]pyrimidin-4-ol 221.0 mg (34.6%)

    Examples 83-87

    [0780] Following the procedure described in Example 78, Step C, using General Procedure B, methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (0.2 g, 8 mmol, 1.0 equiv) was treated with the appropriate corresponding nitrile HetArCN X6 (1.0 equiv) in dioxane (10.0 mL) to produce the Example compounds (83)-(87) shown in Table F.

    TABLE-US-00008 TABLE F Amount Example Structure Name (Yield) 83 [00221]embedded image 2-(1-(2-(Benzyloxy)ethyl)- 1H-imidazol-2-yl)-7- (pyridin-2-yl)thieno[3,2- d]pyrimidin-4-ol 13.0 mg (3.8%) 84 [00222]embedded image 2-(8-Methoxyimidazo[1,5- a]pyridin-3-yl)-7-(pyridin- 2-yl)thieno[3,2- d]pyrimidin-4-ol 7.0 mg (2.2%) 85 [00223]embedded image 2-(7-Methoxyimidazo[1,5- a]pyridin-3-yl)-7-(pyridin- 2-yl)thieno[3,2- d]pyrimidin-4-ol 6.8 mg (2.1%) 86 [00224]embedded image 2-(6-Methoxyimidazo[1,5- a]pyridin-3-yl)-7-(pyridin- 2-yl)thieno[3,2- d]pyrimidin-4-ol 19.0 mg (5.8%) 87 [00225]embedded image 2-(1-(2-Methoxyethyl)-1H- benzo[d]imidazol-2-yl)-7- (pyridin-2-yl)thieno[3,2- d]pyrimidin-4-ol 21.0 mg (6.0%)

    Example 88

    [0781] ##STR00226##

    7-(Pyridin-2-yl)-2-(pyrimidin-4-yl)thieno[3,2-d]pyrimidin-4-ol (88)

    Step A: (E)-3-Ethoxy-2-(pyridin-2-yl)acrylonitrile 78B

    [0782] A mixture of 2-(pyridin-2-yl)acetonitrile 78A (25.0 g, 211.63 mmol), triethoxymethane (47.05 g, 317.45 mmol, 52.86 ml, 1.5 equiv) and acetic anhydride (194.45 g, 1.9 mol) was refluxed for 4 hours. Then it was evaporated and purified by flash chromatography (SiO.sub.2, Hexane-EtOAc as a solvent mixture) to give (E)-3-ethoxy-2-(pyridin-2-yl)acrylonitrile 78B (36.3 g, 3.82% purity, 7.96 mmol, 3.8% yield).

    Step B: Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C

    [0783] To the stirred solution of methyl 2-sulfanylacetate (25.44 g, 239.65 mmol, 21.43 ml, 1.15 equiv) in dry THE (300 mL) sodium methanolate (20.26 g, 375.11 mmol) was added at 0° C. after 5 min to the resulted mixture a solution of (E)-3-ethoxy-2-(pyridin-2-yl)acrylonitrile 78B (36.3 g, 3.82% purity, 208.39 mmol) in dry THF (100 mL) was added dropwise. This mixture was heated at 65° C. overnight. Then the solvent was evaporated, 200 mL of water was added and extracted with EtOAc. The organic extracts were dried over Na.sub.2SO.sub.4, and concentrated in vacuo. Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (3.5 g, 66.0% purity, 9.86 mmol, 4.7% yield) was obtained after column chromatography.

    Step C: 7-(Pyridin-2-yl)-2-(pyrimidin-4-yl)thieno[3,2-d]pyrimidin-4-ol (88)

    [0784] Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (150.0 mg, 640.28 μmol) and pyrimidine-4-carbonitrile (67.37 mg, 641.0 μmol) were dissolved in DMF (20 mL) and sodium hydride (46.15 mg, 1.92 mmol) was added at 0° C. The reaction mixture was heated at 80° C. overnight. The solution of NH.sub.4Cl (15 mL) was added dropwise to reaction mixture after cooling to 0° C., then extracted with EtOAc (3×30 mL). The organic extracts were dried over Na.sub.2SO.sub.4, concentrated in vacuo and purified by HPLC (2-10 min; 40-65% water/MeCN; Flow rate 30 mL/min; loading pump MeCN 4 mL/min; column SunFire 19*100 mm) to give 7-(Pyridin-2-yl)-2-(pyrimidin-4-yl)thieno[3,2-d]pyrimidin-4-ol (88) (10.1 mg, 95.0% purity, 31.22 μmol, 4.9% yield).

    Example 89

    [0785] ##STR00227##

    7-(Pyridin-2-yl)-2-(pyridin-3-yl)thieno[3,2-d]pyrimidin-4-ol (89)

    [0786] Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (prepared as described in Example 88, Step B) (149.89 mg, 639.81 μmol) and pyridine-3-carbonitrile (66.61 mg, 639.81 μmol) were dissolved in DMF (20 mL) and sodium hydride (76.77 mg, 3.2 mmol) was added at 0° C. The reaction mixture was heated at 80° C. overnight. Solution of NH.sub.4Cl (15 mL) was added dropwise to reaction mixture after cooling to 0° C., then extracted with EtOAc (3×30 mL). The organic extracts were dried over Na.sub.2SO.sub.4, concentrated in vacuo and purified by HPLC (2-10 min; 40-65% water/MeCN; Flow rate 30 mL/min; loading pump MeCN 4 mL/min; column SunFire 19*100 mm) to give 7-(pyridin-2-yl)-2-(pyridin-3-yl)thieno[3,2-d]pyrimidin-4-ol (89) (5.9 mg, 95.0% purity, 18.3 μmol, 2.9% yield).

    Example 90

    [0787] ##STR00228##

    2-(1-Methyl-1H-pyrazol-3-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (90)

    [0788] Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (synthesized as described in Example 88, Step B) (149.95 mg, 640.06 μmol) and 1-methyl-1H-pyrazole-3-carbonitrile X8 (68.56 mg, 640.06 μmol) were dissolved in DMF (20 mL) and sodium hydride (102.4 mg, 4.27 mmol) was added at 0° C. Reaction mixture was heated at 80° C. overnight. Solution of NH.sub.4Cl (15 mL) was added dropwise to reaction mixture after cooling to 0° C., then extracted with EtOAc (3×30 mL). The organic extracts were dried over Na.sub.2SO.sub.4, concentrated in vacuo and purified by HPLC (2-10 min 40-65% water-MeCN; flow 30 mL/min (loading pump MeCN 4 mL/min); column SunFire 19×100 mm, 5 um) to give 2-(1-methyl-1H-pyrazol-3-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (90) (17.5 mg, 95.0% purity, 53.74 μmol, 8.4% yield).

    Example 91

    [0789] ##STR00229##

    2-(1-Methyl-1H-pyrazol-5-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (91)

    [0790] Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (synthesized as described in Example 88, Step B) (149.95 mg, 640.06 μmol) and 1-methyl-1I-pyrazole-5-carbonitrile X9 (68.56 mg, 640.06 μmol) were dissolved in DMF (20 mL) and sodium hydride (102.4 mg, 4.27 mmol) was added at 0° C. Reaction mixture was heated at 80° C. overnight. Solution of NH.sub.4Cl (15 mL) was added dropwise to reaction mixture after cooling to 0° C., then extracted with EtOAc (3×30 mL). The organic extracts were dried over Na.sub.2SO.sub.4, concentrated in vacuo and purified by HPLC (2-10 min 34-45% water-MeCN; flow 30 mL/min (loading pump MeCN 4 mL/min); column SunFire 19×100 mm, 5 um) to give 2-(1-methyl-1H-pyrazol-5-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (91) (7.5 mg, 95.0% purity, 23.03 μmol, 3.6% yield).

    Example 92

    [0791] ##STR00230##

    2-(5-Methylthiazol-2-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (92)

    [0792] Methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate 78C (synthesized as described in Example 88, Step B) (200.26 mg, 854.79 μmol) and thiazole-2-carbonitrile X10 (106.13 mg, 854.79 μmol) were dissolved in DMF (20 mL) and sodium hydride (136.75 mg, 5.7 mmol) was added at 0° C. Reaction mixture was heated at 80° C. overnight. Solution of NH.sub.4Cl (15 mL) was added dropwise to reaction mixture after cooling to 0° C., then extracted with EtOAc (3×30 mL). The organic extracts were dried over Na.sub.2SO.sub.4, concentrated in vacuo and purified by HPLC (2-10 min 60-75% water-methanol; flow 30 mL/min (loading pump 4 mL/min methanol), Column Sun Fire C18 100×19 mm, 5 um) to give 2-(5-methylthiazol-2-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (92). (34.0 mg, 95.0% purity, 98.96 μmol, 11.6% yield).

    Example 93

    [0793] ##STR00231##

    2-(1H-Pyrazolo[4,3-b]pyridine-5-yl)-7-(pyridine-2-yl)thieno[3,2-d]pyrimidin-4-ol (93)

    Step A: 1H-Pyrazolo[4,3-b]pyridine-5-carboxamide 93B

    [0794] To a solution of 1H-pyrazolo[4,3-b]pyridine-5-carboxylic acid 93A (5.0 g, 30.64 mmol) in DMF (50 mL), 1-methyl-1H-imidazole (10.06 g, 122.57 mmol, 9.77 ml, 4.0 equiv), ammonium chloride (3.28 g, 61.28 mmol) and chloro-N,N,N,N-tetramethylformamidinium hexafluorophosphate (10.32 g, 36.77 mmol) were added. The reaction mixture was stirred at room temperature overnight. Water was added (50 mL) and precipitate was formed. The mixture was filtered and washed with water. The precipitate was dried on air to give 1H-pyrazolo[4,3-b]pyridine-5-carboxamide 93B (1.17 g, 98.0% purity, 7.07 mmol, 23.1% yield).

    Step B: 1H-Pyrazolo[4,3-b]pyridine-5-carbonitrile 93C

    [0795] To a solution of 1H-pyrazolo[4,3-b]pyridine-5-carboxamide 93B (1.17 g, 7.22 mmol) and triethylamine (2.34 g, 23.11 mmol, 3.22 ml, 3.2 equiv) in THE (25 mL), trifluoroacetyl 2,2,2-trifluoroacetate (2.43 g, 11.55 mmol, 1.62 mL, 1.6 equiv) was added dropwise at 0° C. The reaction mixture was stirred at room temperature overnight. Then, the mixture was concentrated under reduced pressure. Ethyl acetate was added (30 mL) and washed with water (3×10 mL), The organic extracts were dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give 1H-pyrazolo[4,3-b]pyridine-5-carbonitrile 93C (650.0 mg, 98.53% purity, 4.44 mmol, 61.5% yield).

    Step C: 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-b]pyridine-5-carbonitrile 93D

    [0796] To a solution of 1H-pyrazolo[4,3-b]pyridine-5-carbonitrile 93C (200.66 mg, 1.39 mmol) in DMF (20 mL), sodium hydride (83.52 mg, 3.48 mmol) was added at 0° C. portionwise. The resulting mixture was stirred for 30 min following by the addition of [2-(chloromethoxy)ethyl]trimethylsilane (348.54 mg, 2.09 mmol, 370.0 μL, 1.5 equiv). The reaction mixture was stirred at room temperature overnight. Then, saturated solution of ammonium chloride was added at 0° C. (10 mL), extracted with EtOAc (3×15 mL). Combined EtOAc was washed with water (5×10 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-b]pyridine-5-carbonitrile 93D (400.0 mg, 90.0% purity, 1.31 mmol, 94.2% yield).

    Step D: 7-(Pyridin-2-yl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)thieno[3,2-d]pyrimidin-4-ol 93E

    [0797] 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-b]pyridine-5-carbonitrile 93D (199.77 mg, 728.03 μmol) and methyl 3-amino-4-(pyridin-2-yl)thiophene-2-carboxylate (170.56 mg, 728.03 μmol) were dissolved in DMF (20 mL) and sodium hydride (87.36 mg, 3.64 mmol) was added at 0° C. portionwise. The reaction mixture was heated at 80° C. overnight. Then, the solution of NH.sub.4Cl (15 mL) was added dropwise to reaction mixture after cooling to 0° C., then extracted with EtOAc (3×30 mL). The organic extracts were dried over Na.sub.2S04, concentrated in vacuo to give 7-(pyridin-2-yl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)thieno[3,2-d]pyrimidin-4-ol 93E (400.0 mg, 33.43% purity, 280.56 μmol, 38.5% yield).

    Step E: 2-(1H-Pyrazolo[4,3-b]pyridin-5-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (93)

    [0798] To a solution of 7-(pyridin-2-yl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)thieno[3,2-d]pyrimidin-4-ol 93E (400.0 mg, 839.23 μmol) in DCM (2 mL), 2,2,2-trifluoroacetic acid (191.59 mg, 1.68 mmol) was added. The reaction mixture was stirred at room temperature for 2 days. The mixture was concentrated under reduce pressure and purified by HPLC (2-10 min; 30-45% water/MeOH+NH.sub.3; Flow rate 30 mL/min; loading pump MeOH+NH.sub.3 4 mL/min; column SunFire 19*100 mm) to afford 2-(1H-pyrazolo[4,3-b]pyridin-5-yl)-7-(pyridin-2-yl)thieno[3,2-d]pyrimidin-4-ol (93). (4.2 mg, 95.0% purity, 11.52 μmol, 1.4% yield).

    Example 94

    [0799] ##STR00232##

    2-(1-(2-Methoxyethyl)-1H-imidazol-2-yl)-7-(pyridin-4-yl)thieno[3,2-d]pyrimidin-4-ol (94)

    Step A: (E)-3-Ethoxy-2-(pyridin-4-yl)acrylonitrile 94B

    [0800] A mixture of 2-(pyridin-4-yl)acetonitrile 94A (25.0 g, 211.63 mmol), triethoxymethane (47.05 g, 317.45 mmol, 52.86 ml, 1.5 equiv) and acetic anhydride (194.45 g, 1.9 mol) was refluxed for 4 hours. Then it was evaporated and purified by flash chromatography (SiO.sub.2, Hexane-EtOAc as a mobile phase) to give (E)-3-ethoxy-2-(pyridin-4-yl)acrylonitrile 94B (36.3 g, 3.82% purity, 7.96 mmol, 3.8% yield).

    Step B: Methyl 3-amino-4-(pyridin-4-yl)thiophene-2-carboxylate 94C

    [0801] To the stirred solution of methyl 2-sulfanylacetate (25.44 g, 239.65 mmol, 21.43 ml, 1.15 equiv) in dry THE (300 mL) sodium methanolate (20.26 g, 375.11 mmol) was added at 0° C. The resulting mixture was stirred for 5 min following by the dropwise addition of a solution of (E)-3-ethoxy-2-(pyridin-4-yl)acrylonitrile 94B (36.3 g, 3.82% purity, 208.39 mmol) in dry THE (100 mL). This mixture was heated at 65° C. overnight. Then the solvent was evaporated in vacuo, 200 mL of water was added and extracted with EtOAc. The organic extracts were dried over Na.sub.2SO.sub.4, and concentrated in vacuo. Methyl 3-amino-4-(pyridin-4-yl)thiophene-2-carboxylate 94C (3.5 g, 66.0% purity, 9.86 mmol, 4.7% yield) was obtained after column chromatography (SiO.sub.2, Hexane-EtOAc as a mobile phase).

    Step C: 2-(1-(2-Methoxyethyl)-1H-imidazol-2-yl)-7-(pyridin-4-yl)thieno[3,2-d]pyrimidin-4-ol (94)

    [0802] Methyl 3-amino-4-(pyridin-4-yl)thiophene-2-carboxylate 94C (169.71 mg, 724.4 μmol) and 1-(2-methoxyethyl)-1H-imidazole-2-carbonitrile (109.5 mg, 724.4 μmol) were dissolved in DMF (20 mL) and sodium hydride (115.89 mg, 4.83 mmol) was added at 0° C. Reaction mixture was heated at 80° C. overnight. Solution of NH.sub.4Cl (15 mL) was added dropwise to reaction mixture after cooling to 0° C., then extracted with EtOAc (3×30 mL). The organic extracts were dried over Na.sub.2SO.sub.4, concentrated in vacuo and purified by HPLC (2-10 min 0-45% water-MeOH-0.1% NH.sub.4OH; flow 30 mL/min (loading pump MeOH 4 mL/min); column SunFire 19×100 mm, 5 um) to give 2-(1-(2-methoxyethyl)-1H-imidazol-2-yl)-7-(pyridin-4-yl)thieno[3,2-d]pyrimidin-4-ol (94) (12.4 mg, 95.0% purity, 33.33 μmol, 4.6% yield).

    Example 95

    [0803] ##STR00233##

    7-(3-Fluoropyridin-2-yl)-4-methoxy-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine (95)

    Step A: 7-Bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine 95A

    [0804] To a suspension of 7-bromo-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidin-4-ol 30B (prepared as described in Example 30, Step A) (300.26 mg, 964.96 μmol) in phosphoryl trichloride (3.11 g, 20.26 mmol) triethylamine (99.6 mg, 984.26 μmol) was slowly added at room temperature. The resulting mixture was stirred for 8 h at 60° C. After cooling, the excess of phosphoryl chloride was removed under reduced pressure. The residue was mixed with chloroform (10 mL) and sodium bicarbonate aqueous solution (10 ml). The mixture was extracted with chloroform (10 mL), and the organic phase was separated, dried over Na.sub.2SO.sub.4, filtered and evaporated in vacuo to obtain 7-bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine 95A (300.0 mg, 90.0% purity, 814.19 μmol, 84.4% yield).

    Step B: 7-Bromo-4-methoxy-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine 95B

    [0805] Sodium methanolate (73.31 mg, 1.36 mmol) was added to suspension of 7-bromo-4-chloro-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine 95A (300.0 mg, 904.64 μmol) in MeOH (3 mL). The reaction mixture was stirred overnight at room temperature. After the completion of the reaction, the solvent was removed under reduced pressure. The crude product was purified by column chromatography (SiO.sub.2, Hexane/EtOAc (10/1-1/1) as a solvent mixture) to afford 7-bromo-4-methoxy-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine 95B (100.0 mg, 90.0% purity, 275.06 μmol, 30.4% yield) as yellow solid.

    Step C: 7-(3-Fluoropyridin-2-yl)-4-methoxy-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine (95)

    [0806] 7-Bromo-4-methoxy-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine 95B (100.0 mg, 305.62 μmol) and 2-methyl-6-(tributylstannyl)pyridine (128.55 mg, 336.36 μmol) were mixed in DMSO (3 mL) and stirred under an Ar atmosphere for 5 min followed by the addition of Pd(PPh.sub.3).sub.4 (35.46 mg, 30.58 μmol). The reaction mixture was heated under reflux overnight. The mixture was extracted with ethyl acetate (10 ml*2) and brine (10 mL). The organic phase was washed with saturated aqueous solution of NaCl (10 ml), dried over MgSO.sub.4 and concentrated in vacuo. The crude material was purified via HPLC (Agilent 1260 Infinity systems equipped with DAD and mass-detector; Waters Sunfire C18 OBD Prep Column, 100 A, 5 μm, 19 mm*100 mm with SunFire C18 Prep Guard Cartridge, 100 A, 10 μm, 19 mm*10 mm; 2-10 min, 0-60% MeCN—H.sub.2O+TFA flow 30 ml/min (loading pump 4 ml/min H.sub.2O)) to give 7-(3-fluoropyridin-2-yl)-4-methoxy-2-(1-methyl-1H-imidazol-2-yl)thieno[3,2-d]pyrimidine (95) (3.0 mg, 97.0% purity, 8.47 μmol, 2.8% yield).

    Example 96

    [0807] ##STR00234##

    6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1s,3s)-3-methoxycyclobutyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (96)

    Step A: 6-(1-isopropyl-1H-pyrazol-3-yl)-N-((1s,3s)-3-methoxycyclobutyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (96)

    [0808] 4-Chloro-6-(1-isopropyl-1H-pyrazol-3-yl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidine 31G (142.31 mg, 381.66 μmol) (prepared as described in Example 31, Step F) was dissolved in DMSO (3 mL) and N,N-diisopropylethylamine (148.4 mg, 1.15 mmol, 200.0 μL) with corresponding amine (1s,3s)-3-methoxycyclobutan-1-amine hydrochloride (105.04 mg, 763.32 μmol) was added at room temperature. The mixture was heated at 100° C. overnight, cooled and purified by HPLC. 6-(1-Isopropyl-1H-pyrazol-3-yl)-N-((1s,3s)-3-methoxycyclobutyl)-5-methyl-2-(1-methyl-1H-imidazol-2-yl)thieno[2,3-d]pyrimidin-4-amine (96) (12.1 mg, 95.0% purity, 26.27 μmol, 6.9% yield) was obtained as yellow gum after HPLC.

    Purification and Analytical Procedures:

    [0809] The purification of compounds disclosed herein was performed using HPLC (H.sub.2O—MeOH; Agilent 1260 Infinity systems equipped with DAD and mass-detectors. Waters Sunfire C18 OBD Prep Column, 100A, 5 μm, 19 mm×100 mm with SunFire C18 Prep Guard Cartridge, 100 Å, 10 μm, 19 mm×10 mm) The material was dissolved in 0.7 mL DMSO. Flow: 30 mL/min. Purity of the obtained fractions was checked via the analytical LCMS. Spectra were recorded for each fraction as it was obtained straight after chromatography in the solution form. The solvent was evaporated under the N2 flow upon heating to 80° C. On the basis of post-chromatography LCMS analysis fractions were united. Solid fractions were dissolved in 0.5 mL MeOH and transferred into a pre-weighted marked vials. Obtained solutions were again evaporated under the N2 flow upon heating to 80° C. After drying, products were finally characterized by LCMS and .sup.1H NMR.

    [0810] NMR Instrument specifications: Bruker AVANCE DRX 500, Varian UNITY plus 400.

    [0811] LC/MS Instrument specifications: Agilent 1100 Series LC/MSD system with DAD\ELSD and Agilent LC\MSD VL (G1956A), SL (G1956B) mass-spectrometer. Agilent 1200 Series LC/MSD system with DAD\ELSD and Agilent LC\MSD SL (G6130A), SL (G6140A) mass-spectrometer. All the LC/MS data were obtained using positive/negative mode switching. Column Zorbax SB-C18 1.8 μm 4.6×15 mm Rapid Resolution cartridge (PN 821975-932) Mobile phase A-acetonitrile, 0.1% formic acid, B-water (0.1% formic acid) Flow rate 3 ml/min Gradient 0 min-100% B, 0.01 min-100% B, 1.5 min-0% B, 1.8 min-0% B, 1.81 min-100% B. Injection volume 1 μl. Ionization mode atmospheric pressure chemical ionization (APCI). Scan range m/z 80-1000.

    [0812] Table 3 below provides the both the calculated and observed mass spectral data for each of the above-noted exemplified compounds, and also provides the calculated Log P (using ChemDraw Professional version 20.0.0.38).

    TABLE-US-00009 TABLE 3 Mass Spectral Data and Calculated LogP M + H M + H LogP Compound MW (calculated) (observed) (calculated) 1 472 472.2 472.2 3.28 2 472 472.2 472.2 2.64 3 486 486.2 486.2 3.09 4 514 514.2 514.4 4.17 5 514 514.2 514.4 4.17 6 530 530.2 530.1 3.36 7 516 516.2 516.0 2.91 8 373 374.1 374.2 2.87 9 359 360.1 360.2 2.42 10 329 330.1 330.2 3.03 11 292 293.1 293.2 1.60 12 362 363.1 363.2 1.67 13 334 335.1 335.2 2.49 14 350 351.1 351.1 1.68 15 368 369.1 369.2 3.50 16 369 370.1 370.2 2.88 17 401 401.1 401.2 1.37 18 272 273.0 273.0 1.66 19 326 327.1 327.0 2.45 20 314 315.0 315.2 2.55 21 330 331.0 331.2 1.74 22 342 343.3 343.5 1.73 23 348 349.0 349.0 3.56 24 349 350.0 350.0 2.94 25 346 347.1 347.2 2.39 26 320 321.1 321.0 3.14 27 324 325.1 325.0 3.67 30 327 328.1 328.2 4.00 31 452 452.3 452.2 3.57 32 452 452.3 452.2 3.24 33 452 452.2 452.2 2.97 34 438 438.2 438.0 2.97 35 412 412.2 412.2 2.97 36 426 426.2 426.2 3.22 37 424 424.2 424.2 2.59 38 452 452.3 452.2 2.70 39 468 468.2 468.2 2.43 40 452 452.2 452.2 3.46 41 468 468.2 468.2 2.94 42 482 482.2 482.2 3.67 43 482 482.2 482.2 2.96 44 482 482.2 482.0 2.34 45 468 468.2 468.2 2.34 46 450 450.2 450.2 2.34 47 452 452.2 452.2 2.96 48 452 452.2 452.2 3.39 49 452 452.2 452.2 3.02 50 452 452.2 452.2 3.02 51 451 451.2 451.2 3.67 52 451 451.2 451.2 3.67 53 448 448.2 448.2 1.91 54 466 466.2 466.2 1.91 55 466 466.2 466.2 2.84 56 465 465.2 465.0 3.19 57 434 434.2 434.2 3.19 58 462 462.2 462.2 2.01 59 434 434.2 434.2 2.55 60 465 465.2 465.1 2.25 61 452 452.2 452.2 1.91 62 480 480.2 480.3 1.86 63 466 466.2 466.2 3.07 64 496 460.2 460.2 4.12 65 497 461.2 461.1 3.45 66 462 462.2 462.2 4.59 67 461 461.2 461.2 4.68 68 462 462.2 462.2 4.08 69 449 449.2 449.2 3.97 70 449 449.2 449.2 3.88 71 449 449.2 449.4 3.40 72 477 477.2 477.2 4.10 73 506 506.2 506.2 3.89 74 506 506.2 506.2 4.55 75 506 506.2 506.2 3.41 76 514 514.2 514.2 3.89 77 514 514.2 514.2 4.11 78 353 354.1 354.0 4.66 79 367 368.1 368.2 4.88 80 367 368.1 368.0 2.62 81 421 422.1 422.0 3.04 82 382 382.1 382.0 3.27 83 430 430.1 430.0 3.91 84 375 376.1 376.0 3.59 85 375 376.1 376.2 4.66 86 375 376.1 376.0 2.08 87 404 404.1 404.0 2.08 88 307 308.1 308.0 2.08 89 306 307.1 307.0 4.58 90 309 310.1 310.0 2.92 91 310 310.1 310.0 3.09 92 326 327.0 327.0 2.43 93 346 347.1 347.0 2.21 94 353 354.1 354.0 3.28 95 341 342.1 342.0 3.29 96 438 438.2 438.2 2.52

    Kinetic Solubility Assay:

    [0813] Equipment: Water purification system Millipore Milli-Q Gradient A10 (Millipore, France); Thermomixer R Block, 1.5 ml (Eppendorf, Germany; Cat #5355); Matrix Multichannel Electronic Pipette 2-125 μL, 5-250 μL, 15-1250 μL (Thermo Scientific, USA; Cat ##2011, 2012, 2004); SpectraMax Plus Microplate Reader (Molecular Devices, USA; Product #02196); Multi-Well Plate Vacuum Manifold (Pall Corporation, USA; Product #5014); and Vacuum pump (Millipore, USA; Model #XX5500000)

    [0814] Analytical System: The measurements were performed using SpectraMax Plus reader in UV-Vis mode. Acquisition and analysis of the data were performed using SoftMax Pro v.5.4 (Molecular Devices) and Excel 2010 data analysis software.

    [0815] Methods: Briefly, using a 20 mM stock solution of the compound in 100% DMSO dilutions were prepared to a theoretical concentration of 400 μM in duplicates in phosphate-buffered saline pH 7.4 (138 mM NaCl, 2.7 mM KCl, 10 mM K-phosphate) with 2% final DMSO. The experimental compound dilutions in PBS were further allowed to equilibrate at 25° C. on a thermostatic shaker for two hours and then filtered through HTS filter plates using a vacuum manifold. The filtrates of test compounds were diluted 2-fold with acetonitrile with 2% DMSO before measuring.

    [0816] In parallel, compound dilutions in acetonitrile/PBS (1:1) were prepared to theoretical concentrations of 0 μM (blank), 10 μM, 25 μM, 50 μM, 100 μM, and 200 μM with 2% final DMSO to generate calibration curves. Ondansetron was used as reference compound to control proper assay performance. 200 μL of each sample was transferred to 96-well plate and measured in 200-550 nm range with 5 nm step.

    [0817] The concentrations of compounds in PBS filtrate are calculated using a dedicated Microsoft Excel calculation script. Proper absorbance wavelengths for calculations are selected for each compound manually based on absorbance maximums (absolute absorbance unit values for the minimum and maximum concentration points within 0-3 OD range). Each of the final datasets is additionally visually evaluated by the operator and goodness of fit (R2) is calculated for each calibration curve. The effective range of this assay is approximately 2-400 μM and the compounds returning values close to the upper limit of the range may have higher actual solubility (e.g. 5′-deoxy-5-fluorouridine). This method is not suitable for liquid (at 25° C.) substances (were not present among the tested compounds). Table 3A provides the kinetic solubility for some of the exemplified compounds.

    TABLE-US-00010 TABLE 3A Kinetic Solubility Kinetic Solubility (μM) (pH 7.4 PBS + Compound 2% DMSO) 4 5.0 33 49.0 34 >400 36 268.0 38 >400 40 82.0 44 76.0 63 18.0 66 4.0 69 79.0

    BIOLOGICAL EXAMPLES

    Biological Example 1

    Ras GTP Binding Domain Inhibition Assay

    [0818] The following method was developed as specific assay for the following proteins: KRas wild type, KRas Q61H mutant, KRas G12C mutant, KRas G12D mutant, Rac-1, and Rho-A.

    [0819] Buffer-I:

    [0820] 25 mM Tris-HCl, pH 7.4

    [0821] 27 mM KCl

    [0822] 137 mM NaCl

    [0823] 1 mM MgCl.sub.2

    [0824] 1 mM DTT

    [0825] Buffer-II:

    [0826] 50 mM Tris-HCl, pH 7.0

    [0827] 1 mM MgCl.sub.2

    [0828] 1 mM DTT

    [0829] The small GTPases proteins: KRas wild type. KRas Q61H mutant, KRas G12C mutant, KRas G12D mutants, Rac-1, and Rho-A were expressed as His-tagged proteins. In addition, the Guanosine nucleotide Exchange Factor (GEF) Sos protein (residues 556-1049) was expressed as a His-tagged protein. In cells, the guanine nucleotide exchange factor Sos protein promotes activation of Ras proteins by stimulating an exchange of GDP for GTP. The inclusion of Sos to the Ras GTP binding domain inhibition assay may be considered as an alternative representation of physiological cellular conditions for evaluating the inhibitory activity of some of the tested small molecules.

    [0830] For the assay, all purified small GTPases proteins were diluted in Buffer-I or Buffer-II to a final concentration of 10-30 μg/mL. 200 μL of each diluted protein was added to a nickel-coated 96-well plate and incubated overnight at 4° C. Then the protein solution was discarded and 200 μL of Buffer-I or Buffer-II was added to each well in the presence of 1% DMSO. Compounds to be tested were added to the protein-coated wells at final concentration of 20 μM, and incubated for 3 hours at room temperature with and without 10-30 μg/mL of Sos added to the final hour of the incubation. When performing IC.sub.50 measurements a serial dilution of all tested concentrations was added. Then Cy3-GTP or Cy5-GTP was added to each well to a final concentration of 100 nM. The labeled GTP was incubated for 45 minutes at room temperature. Following GTP incubation, wells were washed 3× in Buffer-I or Buffer-II and 200 μL of Buffer-I or Buffer-II was added to each well. Following washes, the amount of bound labeled-GTP was measured using a SpectraMax M3 (Molecular Devices).

    [0831] Table 4 and Table 5 show % inhibition data, and Table 6 shows IC.sub.50 measurements, for selected compounds tested in the screening assays described above.

    TABLE-US-00011 TABLE 4 % Inhibition at 20 μM of K-Ras mutant and wild-type protein KRas wild KRas wild KRas G12D KRas G12D KRas G12C KRas G12C KRas Q61H KRas Q61H type: type + SOS: mutant: mutant + SOS: mutant: mutant + SOS: mutant: mutant + SOS: Compound % inh. % inh. % inh. % inh. % inh. % inh. % inh. % inh. 1 0 A 0 0 A A A A 2 D D C C D D C D 3 C D D D D D D D 4 C D C C C D C D 5 C D A C C D B D 6 D D D D D D D D 7 D D C D D D D D 8 D D D D D D D D 9 D D C D D D D D 10 C D C C D D C D 11 0 A A 0 A A 0 0 12 0 A A 0 A A A 0 13 0 A A 0 0 0 0 0 14 0 0 A A 0 0 0 A 15 A 0 0 0 A 0 A 0 16 A 0 A A A A A 0 17 A A 0 0 0 0 A 0 18 B A C B B B B B 19 A A B A A A A A 20 C A B B C B B B 21 B A B B B B B B 22 A A A 0 A 0 A A 23 A A A A A 0 0 A 24 A A A 0 A A A 0 25 0 0 0 0 0 0 0 0 26 D C D D D D D D 27 D D C D C D C D 30 D D C C C D C D 31 B D B D C D B D 32 D D B D C D C D 33 D D C D D D D D 34 D D D D D D D D 35 D D D D D D D D 36 D D C D D D D D 37 D D D D D D D D 38 D D C D C D D D 39 D C C D C C C D 40 D D B B C D C D 41 C C 0 B C C C B 42 C D D C D D D D 43 B D D D A D A D 44 D D C D D D D D 45 D D D D C D D D 46 D C B D C C D D 47 D D C C C D C D 48 D C B C C C C C 49 C D 0 B A C C C 50 C C 0 A B C B C 51 C C B C C B C C 52 C C C C C B C C 53 B C A B B B B B 54 D D C D D D C D 55 A A C B B B A B 56 D D B D D D C D 57 B B A B B B B B 58 D D B C D D C C 59 B D N/T N/T C D C D 60 C C N/T N/T B C C C 61 D D N/T N/T C D D D 62 C C B C N/T C D C 63 C B C C B B C B 64 0 D N/T N/T B D B D 65 B C B C B D B C 66 C C D C B B C B 67 B C 0 C B C C C 68 D D D D D C D C 69 C C C C C C D D 70 B B B B C B B B 71 B B 0 A B B B C 72 A 0 C 0 0 A 0 0 73 A D N/T N/T A D C C 74 B D N/T N/T A C C C 75 B C N/T N/T B C C B 76 C C C B A 0 B C 77 B B C B B 0 A B 78 D C D C C C D C 79 D C C C C C C C 80 D C D D C C D C 81 D D C D C C C C 82 C C C C C C C C 83 C B C C A C B C 84 B A B B B A B B 85 C A C B B A C B 86 C B C C B B B C 87 B A C B A A A B 88 B A C B A B B B 89 0 0 0 0 0 0 A A 90 0 0 B A A A A B 91 0 0 A A 0 A A B 92 C C D C C C D C 93 B 0 B A B A A A 94 C C D D C C C C 95 D D D D D D D D 96 0 A 0 A 0 0 0 0 0 = 0% inhibition, A = 1-25% inhibition, B = 26-50% inhibition, C = 51-75% inhibition, D = 76-100% inhibition, N/T = Not tested.

    TABLE-US-00012 TABLE 5 % Inhibition at 20 μM of KRas G12D mutant, Rac-1 and Rho-A protein KRas G12D KRas G12D mutant: mutant + SOS: Rac-1: Rac-1 + SOS: Rho-A: Rho-A + SOS: Compound % inh. % inh. % inh. % inh. % inh. % inh. 1 0 0 0 A A 0 2 C C C C D D 3 D D D D C D 4 C C A B B B 5 A C B B C C 6 D D D D D D 7 C D D D D D 8 D D D D D D 9 C D D D D D 10 C C D D D D 11 A 0 0 0 A A 12 A 0 0 A A A 13 A 0 A 0 A A 14 A A 0 0 A A 15 0 0 0 0 0 0 16 A A A 0 0 A 17 0 0 0 A B 0 18 C B B B B B 19 B A A A B B 20 B B B B B B 21 B B B B C B 22 A 0 A A A A 23 A A A A A A 24 A 0 0 A A A 25 0 0 0 0 0 0 26 D D D D D D 27 C D D D D D 30 C C C C D D 31 B D N/T B C C 32 B D B B C C 33 C D C C D D 34 D D D D D D 35 D D D D D D 36 C D D D B D 37 D D D D D D 38 C D D D D D 39 C D C C D D 40 B B C C D D 41 0 B D D C D 42 D C C C C D 43 D D B C D D 44 C D D D D D 45 D D D D D D 46 B D D D D D 47 C C C C D D 48 B C B B C D 49 0 B B N/T C N/T 50 0 A 0 N/T D N/T 51 B C C N/T D N/T 52 C C C N/T D N/T 53 A B A N/T D N/T 54 C D B N/T C N/T 55 C B 0 N/T C N/T 56 B D D N/T D N/T 57 A B 0 N/T C N/T 58 B C B N/T D N/T 59 N/T N/T C N/T C N/T 60 N/T N/T C N/T D N/T 61 N/T N/T B N/T C N/T 62 B C A N/T D N/T 63 C C B N/T C N/T 64 N/T N/T B N/T B N/T 65 B C B N/T C N/T 66 D C C N/T D N/T 67 0 C A N/T B N/T 68 D D D N/T D N/T 69 C C A B C C 70 B B B B C C 71 0 A 0 N/T D N/T 72 C 0 0 N/T A N/T 73 N/T N/T A N/T C N/T 74 N/T N/T A N/T C N/T 75 N/T N/T A N/T C N/T 76 C B A N/T B N/T 77 C B A N/T B N/T 78 D C C N/T C N/T 79 C C C N/T D N/T 80 D D D N/T C N/T 81 C D D N/T D N/T 82 C C D N/T C N/T 83 C C D N/T C N/T 84 B B B N/T C N/T 85 C B C N/T C N/T 86 C C C N/T C N/T 87 C B C N/T C N/T 88 C B C N/T C N/T 89 0 0 0 N/T A N/T 90 B A B N/T C N/T 91 A A 0 N/T 0 N/T 92 D C D N/T D N/T 93 B A A N/T C N/T 94 D D D N/T D N/T 95 D D D D D D 96 0 A A A 0 0 0 = 0% inhibition, A = 1-25% inhibition, B = 26-50% inhibition, C = 51-75% inhibition, D = 76-100% inhibition, N/T = Not tested.

    TABLE-US-00013 TABLE 6 KRas 061H mutant IC.sub.50 (μM) values KRas Q61H mutant + SOS: Compound IC50 (μM) 1 I 2 K 3 J 4 K 5 K 6 K 7 K 8 K 9 J 10 J 11 I 12 I 13 I 14 I 15 I 16 I 17 I 18 I 19 I 20 I 21 I 22 I 23 I 24 I 25 N/T 26 J 27 K 30 K 31 K 32 K 96 I I = >10 uM IC.sub.50, J = 5-10 uM IC.sub.50, K = 1-4.99 uM IC.sub.50, L = <1 uM IC.sub.50, N/T = Not tested

    Biological Example 2

    Protocols for Cell-Based Phosphorylation Assays

    Protocol for Western ERK1/2 Cell-Based Phosphorylation Assay

    [0832] Cell lines: Human tumor-derived pancreatic cancer cell line Panc-1 was purchased from American Type Culture Collection and grown in complete DMEM-High Glucose, supplemented with penicillin (100 U/mL), streptomycin (100 μg/mL), and 10% heat-inactivated FBS at 37° C. in a humidified incubator with 5% CO.sub.2.

    [0833] Method: Cells were plated at 350000 cells/well density in a 12-well plate, allowed 3 hours to adhere to the plate, then starved in the appropriate medium in the presence of 0.5% FBS overnight. The small molecules to be tested were added to the cells in the final concentration of 10 μM in the presence of 0.3% DMSO for 6 hours incubation at 37° C. For IC.sub.50 value determination, serial dilutions of compounds were added to cells under the same conditions. Next, cells were stimulated with 1.5 ng/ml EGF for 15 minutes then cells were lysed with lysis buffer containing 1% Triton X-100, EDTA, and Halt™ Protease & Phosphatase Inhibitor Cocktail (Thermo Scientific). Protein concentration was assessed by BCA protein assay (Thermo Scientific). Phosphorylation level of ERK1/2 was determined by western blot.

    [0834] Western blot protocol: Equal amounts of protein (15-50 μg) were separated by SDS-PAGE and transferred to nitrocellulose membranes (Invitrogen by Thermo Fisher Scientific). The membrane was stained with Ponceau S Stain (Boston BioProducts) to verify uniform protein loading. Membranes were blocked with 10% milk and phosphorylation levels of ERK1/2 were assessed by incubating overnight at 4° C. with anti-phospho-p44/42 (Thr202/Tyr204) antibody (Cell Signaling) followed by HRP-conjugated secondary antibody (Jackson Immunoresearch, West Grove, Pa.). Bands were incubated in Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare) and visualized using the ChemiDoc MP imaging system (Bio-Rad). Protocol for Western AKT cell-based phosphorylation assay

    Human Tumor-Derived Pancreatic Cancer Cell Line Panc-1 was Purchased from

    [0835] American Type Culture Collection and grown in complete DMEM-High Glucose supplemented with penicillin (100 U/mL), streptomycin (100 μg/mL), and 10% heat-inactivated FBS at 37° C. in a humidified incubator with 5% CO.sub.2.

    [0836] Method: Cells were plated at 350000 cells/well density in a 12-well plate, allowed 3 hours to adhere to the plate, then starved in the appropriate medium in the presence of 0.5% FBS overnight. The small molecules to be tested were added to the cells in the final concentration of 10 μM in the presence of 0.3% DMSO for 6 hours incubation at 37° C. Next, cells were stimulated with 1.5 ng/ml EGF for 15 minutes then cells were lysed with lysis buffer containing 1% Triton X-100, EDTA, and Halt™ Protease & Phosphatase Inhibitor Cocktail (Thermo Scientific). Protein concentration was assessed by BCA protein assay (Thermo Scientific). Phosphorylation level of Akt was determined by western blot.

    [0837] Western blot protocol: Equal amounts of protein (15-50 μg) were separated by SDS-PAGE and transferred to nitrocellulose membranes (Invitrogen by Thermo Fisher Scientific). The membrane was stained with Ponceau S Stain (Boston BioProducts) to verify uniform protein loading. Membranes were blocked with 10% milk and phosphorylation levels of Akt were assessed by incubating overnight at 4° C. with anti-phospho-Akt (Ser473) antibody (Cell Signaling) followed by HRP-conjugated secondary antibody (Jackson Immunoresearch, West Grove, Pa.). Bands were incubated in Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare) and visualized using the ChemiDoc MP imaging system (Bio-Rad).

    [0838] Table 7 shows inhibition data for selected compounds tested in one or more of the cellular assays described above.

    TABLE-US-00014 TABLE 7 % Inhibition of ERK1/2 phosphorylation (at 10 μM) and % Inhibition of AKT phosphorylation (at 10 μM) in PANC-1 pancreatic cancer cell line % inh of ERK1/2 % inh of AKT phosphorylation phosphorylation Compound in PANC-1 in PANC-1 1 0 0 2 C 0 3 A 0 4 D D 5 D D 6 D 0 7 D 0 8 0 0 9 0 0 10 A 0 11 0 0 12 0 0 13 0 0 14 0 0 15 0 0 16 0 0 17 A 0 18 0 0 19 0 0 20 0 0 21 0 0 22 0 0 23 0 0 24 0 0 25 0 0 26 0 0 30 0 0 31 D 0 32 D 0 33 D 0 34 D 0 35 0 0 36 C 0 37 0 0 38 D B 39 0 <B> 40 D D 41 0 0 42 B <D> 43 B <D> 44 D <A> 45 B <D> 46 0 A 47 D 0 48 D 0 49 D D 50 D D 51 0 0 52 0 0 53 0 0 54 D <A> 55 D <A> 56 0 0 57 A 0 58 A 0 59 0 0 60 0 0 61 D 0 62 D B 63 D C 64 D B 65 D B 66 0 B 67 0 <A> 68 C <A> 69 D D 70 0 0 71 D D 72 D B 73 D A 74 D 0 75 D 0 76 D 0 77 B <A> 78 0 0 79 0 <B> 80 A <D> 81 B <D> 82 A <D> 83 B <D> 84 0 A 85 0 0 86 0 B 87 A <A> 88 <B> <B> 89 A 0 90 0 0 91 0 0 92 0 0 94 0 0 95 0 0 96 B 0 0 = 0% inhibition, A = 1-25% inhibition, B = 26-50% inhibition, C = 51-75% inhibition, D = 76-100% inhibition. Values in brackets “< >” are understood to be activation %, rather than inhibition.

    Biological Example 3

    Protocol for Cell Proliferation Assay

    [0839] Cell lines: Human tumor-derived cell lines were purchased from American Type Culture Collection. Human tumor-derived pancreatic cancer cell line MIA-PACA2 (having a KRAS G12C mutation) and Panc-1 (having a KRAS G12D mutation), and Human non-small cell lung cancer (NSCLC) cell line A549 (having a KRAS G12S mutation) were grown in complete DMEM-High Glucose. Human colon cancer cell line SW-620 (having a KRAS G12V mutation) was grown in RPMI. All cell lines were supplemented with penicillin (100 U/mL), streptomycin (100 μg/mL), and 10% heat-inactivated FBS at 37° C. in a humidified incubator with 5% CO.sub.2.

    [0840] Method: Cells were plated at 2000 to 5000 cells/well density in 96-wells plate and cultured overnight. Small molecules to be tested were added to the cells in the final concentration of 10 μM in the presence of 0.3% DMSO and 10% FBS, and incubated for 2-4 days at 37° C. in a humidified incubator with 5% CO.sub.2. For IC.sub.50 value determination, serial dilutions of compounds were added to cells under the same conditions.

    [0841] Assay: At the end of the incubation period, cell viability was measured using the CellTiter 96® Aqueous One Solution Cell Proliferation Assay according to manufacturer specifications (Promega, Madison, Wis.). The percentage of small molecules inhibition of cellular proliferation was calculated from raw data.

    [0842] Table 8 shows inhibition data for selected compounds tested in the MIA-PaCa-2 pancreatic cancer cell line using the cellular assay conditions described above.

    TABLE-US-00015 TABLE 8 % Inhibition of cell proliferation at 10 μM in MIA-PaCa-2 pancreatic cancer cell line with select IC.sub.50 (μM) values in the MIA PaCa-2 pancreatic cancer cell line % inh cell proliferation IC.sub.50 (μM) cell proliferation Compound (10 μM) in MIA PaCa-2 in MIA PaCa-2 1 0 I 2 C K 3 D K 4 D L 5 D L 6 D K 7 D K 8 B I 9 A I 10 B I 11 A I 12 A I 13 0 I 14 0 I 15 A I 16 A I 17 A I 18 0 I 19 0 I 20 A I 21 A I 22 A I 23 0 I 24 A I 25 A I 26 C J 27 D K 30 B I 31 D L 32 D L 33 D K 34 D K 35 D J 36 D L 37 C J 38 D K 39 D J 40 D L 41 C J 42 C J 43 C J 44 D K 45 B J 46 A I 47 D L 48 D L 49 D J 50 D A 51 0 I 52 A I 53 D J 54 D L 55 D L 56 A I 57 C J 58 D J 59 C J 60 A I 61 D L 62 D L 63 D L 64 D J 65 D J 66 D L 67 D J 68 D K 69 D J 70 D J 71 D J 72 C J 73 D K 74 D L 75 D J 76 D L 77 D J 78 C J 79 D K 80 D K 81 D K 82 D K 83 D J 84 C J 85 C J 86 C J 87 D J 88 B I 89 B I 90 A I 91 0 I 92 0 I 94 C J 95 0 I 96 A I AMG510 (sotorasib) N/T L 0 = 0% inhibition, A = 1-25% inhibition, B = 26-50% inhibition, C = 51-75% inhibition, D = 76-100% inhibition; I = >10 μM IC.sub.50, J = 5-10 μM IC.sub.50, K = 1-4.99 μM IC.sub.50, L = < 1 μM IC.sub.50; N/T-Not tested

    [0843] Table 9 shows inhibition data (IC50 (μM) values-micromolar units) for selected compounds tested in the MIA-PaCa-2 pancreatic cancer cell line, the PANC-1 pancreatic cancer cell line, the A-549 NSCLC cell line, and the SW-620 colorectal cancer cell line using the cellular assay conditions described above.

    TABLE-US-00016 TABLE 9 % Inhibition of cell proliferation-IC.sub.50 (μM) values in the MIA-PaCa-2 pancreatic cancer cell line, the PANC-1 pancreatic cancer cell line, the A-549 NSCLC cell line, and the SW-620 colorectal cancer cell line IC.sub.50 (μM) cell IC.sub.50 (μM) cell IC.sub.50 (μM) cell IC.sub.50 (μM) cell proliferation in proliferation in proliferation in proliferation in Compound MIA PaCa-2 PANC-1 A549 SW-620  4 M M L L 38 K K K K 63 M L L M 87 I M N/T M AMG510 (sotorasib) M I I I I = >3 μM IC.sub.50, J = 1-3 μM IC.sub.50, K = <1-0.75 μM IC.sub.50, L = <0.75-0.40 μM IC.sub.50, M = <0.40 μM IC.sub.50; N/T-Not tested.

    EXEMPLARY EMBODIMENTS

    [0844] Embodiment 1. A compound of Formula IA:

    ##STR00235##

    [0845] or a pharmaceutically acceptable derivative thereof, wherein:

    [0846] —NR.sup.1R.sup.2 is

    ##STR00236##

    [0847] R.sup.3 is hydrogen, —CH.sub.3, —CF.sub.3, or phenyl;

    [0848] R.sup.4 is hydrogen,

    ##STR00237##

    and

    [0849] R.sup.5 is —CH.sub.3 or —CH.sub.2CF.sub.3.

    [0850] Embodiment 2. The compound or pharmaceutically acceptable derivative of embodiment 1, wherein —NR.sup.1R.sup.2 is:

    ##STR00238##

    [0851] Embodiment 3. The compound or pharmaceutically acceptable derivative of embodiment 1 or 2, wherein R.sup.3 is hydrogen.

    [0852] Embodiment 4. The compound or pharmaceutically acceptable derivative of embodiment 1 or 2, wherein R.sup.3 is —CH.sub.3.

    [0853] Embodiment 5. The compound or pharmaceutically acceptable derivative of embodiment 1 or 2, wherein R.sup.3 is phenyl.

    [0854] Embodiment 6. The compound or pharmaceutically acceptable derivative of embodiment 1, wherein:

    [0855] —NR.sup.1R.sup.2 is

    ##STR00239##

    [0856] R.sup.3 is hydrogen or phenyl;

    [0857] R.sup.4 is hydrogen or

    ##STR00240##

    and

    [0858] R.sup.5 is —CH.sub.3.

    [0859] Embodiment 7. The compound or pharmaceutically acceptable derivative of embodiment 6, wherein R.sup.3 is hydrogen.

    [0860] Embodiment 8. The compound or pharmaceutically acceptable derivative of embodiment 6, wherein R.sup.3 is phenyl.

    [0861] Embodiment 9. The compound or pharmaceutically acceptable derivative of any one of embodiments 1-8, wherein R.sup.4 is hydrogen.

    [0862] Embodiment 10. The compound or pharmaceutically acceptable derivative of any one of embodiments 1-8, wherein R.sup.4 is:

    ##STR00241##

    [0863] Embodiment 11. The compound or pharmaceutically acceptable derivative of any one of embodiments 1-10, wherein the compound of Formula IA is:

    ##STR00242##

    [0864] Embodiment 12. A compound of Formula IB:

    ##STR00243##

    [0865] or a pharmaceutically acceptable derivative thereof, wherein:

    [0866] R.sup.6 is

    ##STR00244##

    [0867] —NR.sup.7R.sup.8 is

    ##STR00245##

    [0868] R.sup.9 is hydrogen, —CH.sub.3, —CF.sub.3, or phenyl; and

    [0869] R.sup.10 is

    ##STR00246##

    [0870] Embodiment 13. The compound or pharmaceutically acceptable derivative of embodiment 12, wherein R.sup.6 is:

    ##STR00247##

    [0871] Embodiment 14. The compound or pharmaceutically acceptable derivative of embodiment 12, wherein R.sup.6 is:

    ##STR00248##

    [0872] Embodiment 15. The compound or pharmaceutically acceptable derivative of embodiment 12, wherein R.sup.6 is:

    ##STR00249##

    [0873] Embodiment 16. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is:

    ##STR00250##

    [0874] Embodiment 17. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is:

    ##STR00251##

    [0875] Embodiment 18. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is:

    ##STR00252##

    [0876] Embodiment 19. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is:

    ##STR00253##

    [0877] Embodiment 20. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is:

    ##STR00254##

    [0878] Embodiment 21. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is:

    ##STR00255##

    [0879] Embodiment 22. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is: CH.sub.3O CH.sub.3O CH.sub.3O

    ##STR00256##

    [0880] Embodiment 23. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-15, wherein —NR.sup.7R.sup.8 is:

    ##STR00257##

    [0881] Embodiment 24. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-23, wherein R.sup.9 is hydrogen.

    [0882] Embodiment 25. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-23, wherein R.sup.9 is —CH.sub.3.

    [0883] Embodiment 26. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-23, wherein R.sup.9 is phenyl.

    [0884] Embodiment 27. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-26, wherein R.sup.10 is:

    ##STR00258##

    [0885] Embodiment 28. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-26, wherein R.sup.10 is:

    ##STR00259##

    [0886] Embodiment 29. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-26, wherein R.sup.10 is:

    ##STR00260##

    [0887] Embodiment 30. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-26 wherein R.sup.10 is:

    ##STR00261##

    [0888] Embodiment 31. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-30, wherein the compound of Formula IB is:

    ##STR00262## ##STR00263##

    [0889] Embodiment 32. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-31, wherein the compound of Formula IB is:

    ##STR00264##

    [0890] Embodiment 33. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-32, wherein the compound of Formula IB is:

    ##STR00265##

    [0891] Embodiment 34. The compound or pharmaceutically acceptable derivative of embodiment 12, wherein:

    [0892] R.sup.6 is

    ##STR00266##

    [0893] —NR.sup.7R.sup.8 is

    ##STR00267##

    [0894] R.sup.9 is phenyl; and

    [0895] R.sup.10 is

    ##STR00268##

    [0896] Embodiment 35. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-30 or embodiment 34, wherein —NR.sup.7R.sup.8 is:

    ##STR00269##

    [0897] Embodiment 36. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-30 or embodiment 34, wherein —NR.sup.7R.sup.8 is:

    ##STR00270##

    [0898] Embodiment 37. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-30 or embodiments 34-36, wherein R.sup.10 is:

    ##STR00271##

    [0899] Embodiment 38. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-30 or embodiments 34-36, wherein R.sup.10 is:

    ##STR00272##

    [0900] Embodiment 39. The compound or pharmaceutically acceptable derivative of any one of embodiments 12-31 or embodiments 34-38, wherein the compound of Formula IB is:

    ##STR00273##

    [0901] Embodiment 40. A compound of Formula IC:

    ##STR00274##

    [0902] or a pharmaceutically acceptable derivative thereof, wherein:

    [0903] —NR.sup.11R.sup.12 is

    ##STR00275## ##STR00276##

    and

    [0904] R.sup.13 is

    ##STR00277##

    [0905] Embodiment 41. The compound or pharmaceutically acceptable derivative of embodiment 40, wherein R.sup.13 is:

    ##STR00278##

    [0906] Embodiment 42. The compound or pharmaceutically acceptable derivative of embodiment 40, wherein R.sup.13 is:

    ##STR00279##

    [0907] Embodiment 43. The compound or pharmaceutically acceptable derivative of embodiment 40, wherein R.sup.13 is:

    ##STR00280##

    [0908] Embodiment 44. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43, wherein —NR.sup.11R.sup.12 is:

    ##STR00281##

    [0909] Embodiment 45. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-44, wherein —NR.sup.11R.sup.12 is:

    ##STR00282##

    [0910] Embodiment 46. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-44, wherein —NR.sup.11R.sup.12 is:

    ##STR00283##

    [0911] Embodiment 47. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-44, wherein —NR.sup.11R.sup.12 is:

    ##STR00284##

    [0912] Embodiment 48. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-44, wherein —NR.sup.11R.sup.12 is:

    ##STR00285##

    [0913] Embodiment 50. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-44, wherein —NR.sup.11R.sup.12 is:

    ##STR00286##

    [0914] Embodiment 51. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43, wherein —NR.sup.11R.sup.12 is:

    ##STR00287## ##STR00288##

    [0915] Embodiment 52. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00289##

    [0916] Embodiment 53. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00290##

    [0917] Embodiment 54. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00291##

    [0918] Embodiment 55. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00292##

    [0919] Embodiment 56. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00293##

    [0920] Embodiment 57. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00294##

    [0921] Embodiment 58. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00295##

    [0922] Embodiment 59. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00296##

    [0923] Embodiment 60. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-43 or embodiment 51, wherein —NR.sup.11R.sup.12 is:

    ##STR00297##

    [0924] Embodiment 61. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-60, wherein the compound of Formula IC is:

    ##STR00298## ##STR00299## ##STR00300## ##STR00301##

    [0925] Embodiment 62. The compound or pharmaceutically acceptable derivative of any one of embodiments 40-61, wherein the compound of Formula 1C is:

    ##STR00302## ##STR00303## ##STR00304##

    [0926] Embodiment 63. A compound of Formula ID:

    ##STR00305##

    [0927] or a pharmaceutically acceptable derivative thereof, wherein:

    [0928] —NR.sup.14R.sup.15 is

    ##STR00306##

    [0929] Embodiment 64. The compound of embodiment 63, wherein —NR.sup.14R.sup.15 is:

    ##STR00307##

    [0930] Embodiment 65. The compound or pharmaceutically acceptable derivative of embodiment 63, wherein —NR 4R.sup.15 is:

    ##STR00308##

    [0931] Embodiment 66. The compound or pharmaceutically acceptable derivative of embodiment 63, wherein —NR.sup.14R.sup.15 is:

    ##STR00309##

    [0932] Embodiment 67. The compound of embodiment 63, wherein —NR.sup.14R.sup.15 is:

    ##STR00310##

    [0933] Embodiment 68. The compound or pharmaceutically acceptable derivative of any one of embodiments 63-67, wherein R.sup.16 is:

    ##STR00311##

    [0934] Embodiment 69. The compound or pharmaceutically acceptable derivative of any one of embodiments 63-67, wherein R.sup.16 is:

    ##STR00312##

    [0935] Embodiment 70. The compound or pharmaceutically acceptable derivative of any one of embodiments 63-67, wherein R.sup.16 is:

    ##STR00313##

    [0936] Embodiment 71. The compound or pharmaceutically acceptable derivative of embodiment 63, wherein:

    [0937] —NR.sup.14R.sup.15 is

    ##STR00314##

    and

    [0938] R.sup.16 is

    ##STR00315##

    [0939] Embodiment 72. The compound or pharmaceutically acceptable derivative of any one of embodiments 63-71, wherein —NR.sup.14R.sup.15 is

    ##STR00316##

    [0940] Embodiment 73. The compound or pharmaceutically acceptable derivative of any one of embodiments 64-72, wherein the compound of Formula ID is:

    ##STR00317##

    [0941] Embodiment 74. A compound of Formula IE:

    ##STR00318##

    [0942] or a pharmaceutically acceptable derivative thereof, wherein:

    [0943] —NR.sup.17R.sup.18 is

    ##STR00319##

    [0944] Embodiment 75. The compound or pharmaceutically acceptable derivative of embodiment 74, wherein —NR.sup.17R.sup.18 is:

    ##STR00320##

    [0945] Embodiment 76. The compound or pharmaceutically acceptable derivative of embodiment 74, wherein —NR.sup.17R.sup.18 is:

    ##STR00321##

    [0946] Embodiment 77. The compound or pharmaceutically acceptable derivative of embodiment 74, wherein —NR.sup.17R.sup.18 is:

    ##STR00322##

    [0947] Embodiment 78. The compound or pharmaceutically acceptable derivative of embodiment 74, wherein —NR.sup.17R.sup.18 is:

    ##STR00323##

    [0948] Embodiment 79. The compound or pharmaceutically acceptable derivative of embodiment 74, wherein the compound of Formula IE is:

    ##STR00324##

    [0949] Embodiment 80. A compound of Formula IF:

    ##STR00325##

    [0950] or a pharmaceutically acceptable derivative thereof, wherein:

    [0951] —NR.sup.19R.sup.20 is —NH.sub.2,

    ##STR00326##

    [0952] —NR.sup.21R.sup.22 is

    ##STR00327##

    and

    [0953] R.sup.23 is hydrogen or

    ##STR00328##

    [0954] Embodiment 81. The compound or pharmaceutically acceptable derivative of embodiment 80, wherein R.sup.23 is hydrogen.

    [0955] Embodiment 82. The compound or pharmaceutically acceptable derivative of embodiment 80, wherein R.sup.23 is:

    ##STR00329##

    [0956] Embodiment 83. The compound or pharmaceutically acceptable derivative of any one of embodiments 80-82, wherein the compound of Formula IF is:

    ##STR00330## ##STR00331##

    [0957] Embodiment 84. A compound of Formula IIA:

    ##STR00332## [0958] or a pharmaceutically acceptable derivative thereof, wherein:

    [0959] —NR.sup.24R.sup.5 is —NH.sub.2,

    ##STR00333##

    [0960] Embodiment 85. The compound or pharmaceutically acceptable derivative of embodiment 84, wherein the compound of Formula IIA is:

    ##STR00334## ##STR00335##

    [0961] Embodiment 86. A compound of Formula IIB:

    ##STR00336##

    [0962] or a pharmaceutically acceptable derivative thereof, wherein:

    [0963] R.sup.26 is

    ##STR00337## ##STR00338##

    [0964] R.sup.27 is hydrogen, —CH.sub.3, or —CF.sub.3; and

    [0965] R.sup.28 is

    ##STR00339##

    [0966] Embodiment 87. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00340##

    [0967] Embodiment 88. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00341##

    [0968] Embodiment 89. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00342##

    [0969] Embodiment 90. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00343##

    [0970] Embodiment 91. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00344## ##STR00345##

    [0971] Embodiment 92. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00346##

    [0972] Embodiment 93. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00347##

    [0973] Embodiment 94. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00348##

    [0974] Embodiment 95. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00349##

    [0975] Embodiment 96. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein R.sup.26 is:

    ##STR00350##

    [0976] Embodiment 97. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-96, wherein R.sup.27 is hydrogen.

    [0977] Embodiment 98. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-96, wherein R.sup.27 is —CH.sub.3.

    [0978] Embodiment 99. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-96, wherein R.sup.27 is —CF.sub.3.

    [0979] Embodiment 100. The compound or pharmaceutically acceptable derivative of any one of any one of embodiments 86-98, wherein R.sup.28 is:

    ##STR00351##

    [0980] Embodiment 101. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-98, wherein R.sup.28 is:

    ##STR00352##

    [0981] Embodiment 102. The compound or pharmaceutically acceptable derivative of embodiment 86, wherein:

    [0982] R.sup.26 is

    ##STR00353##

    [0983] R.sup.27 is hydrogen; and

    [0984] R.sup.28 is

    ##STR00354##

    [0985] Embodiment 103. The compound or pharmaceutically acceptable derivative of embodiment 102, wherein R.sup.26 is:

    ##STR00355##

    [0986] Embodiment 104. The compound or pharmaceutically acceptable derivative of embodiment 102, wherein R.sup.26 is:

    ##STR00356##

    [0987] Embodiment 105. The compound or pharmaceutically acceptable derivative of any one of embodiments 102-104, wherein R.sup.28 is:

    ##STR00357##

    [0988] Embodiment 106. The compound or pharmaceutically acceptable derivative of any one of embodiments 102-104, wherein R.sup.28 is:

    ##STR00358##

    [0989] Embodiment 107. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-106, with the proviso that when:

    [0990] R.sup.26 is

    ##STR00359##

    and

    [0991] R.sup.27 is hydrogen; then

    [0992] R.sup.28 is not

    ##STR00360##

    [0993] Embodiment 108. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-107, wherein the compound of Formula IIB is:

    ##STR00361## ##STR00362## ##STR00363##

    [0994] Embodiment 109. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-107, wherein the compound of Formula IIB is:

    ##STR00364## ##STR00365## ##STR00366##

    [0995] Embodiment 110. The compound or pharmaceutically acceptable derivative of any one of embodiments 86-107, wherein the compound of Formula IIB is:

    ##STR00367##

    [0996] Embodiment 111. A compound or a pharmaceutically acceptable derivative thereof, wherein the compound is:

    ##STR00368##

    [0997] Embodiment 112. A compound of selected from the group consisting of compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, and 95, or a pharmaceutically acceptable derivative thereof.

    [0998] Embodiment 113. A compound of selected from the group consisting of compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, and 27, or a pharmaceutically acceptable derivative thereof.

    [0999] Embodiment 114. A compound of selected from the group consisting of compounds 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, and 39 or a pharmaceutically acceptable derivative thereof.

    [1000] Embodiment 115. A compound which binds to the GTP binding domain of one or more members of the Ras superfamily and inhibits the one or more members of the Ras superfamily with an IC.sub.50 value of less than 10 micromolar, wherein the compound is the compound or pharmaceutically acceptable derivative of any one of embodiments 1-114.

    [1001] Embodiment 116. The compound of embodiment 115, wherein one or more members of the Ras superfamily is Ras.

    [1002] Embodiment 117. The compound of embodiment 115, wherein one or more members of the Ras superfamily is Rho.

    [1003] Embodiment 118. The compound of embodiment 115, wherein one or more members of the Ras superfamily is Rac.

    [1004] Embodiment 119. The compound of embodiment 116, wherein the Ras is DIRAS1; DIRAS2; DIRAS3; ERAS; GEM; HRAS; KRAS; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS; or RRAS2.

    [1005] Embodiment 120. The compound of embodiment 119, wherein the Ras is HRAS, KRAS, NRAS, or a mutant thereof.

    [1006] Embodiment 121. The compound of embodiment 120, wherein the Ras is HRAS or a mutant thereof.

    [1007] Embodiment 122. The compound of embodiment 120, wherein the Ras is KRAS or a mutant thereof.

    [1008] Embodiment 123. The compound of embodiment 120, wherein the Ras is NRAS or a mutant thereof.

    [1009] Embodiment 124. The compound of embodiment 117, wherein the Rho is RHOA; RHOB; RHOBTB1; RHOBTB2; RHOBTB3; RHOC; RHOD; RHOF; RHOG; RHOH; RHOJ; RHOQ; RHOU; RHOV; RND1; RND2; RND3; RAC1; RAC2; RAC3; CDC42, or a mutant thereof.

    [1010] Embodiment 125. The compound of embodiment 117, wherein the Rho is Rac.

    [1011] Embodiment 126. The compound of embodiment 118 or 125, wherein the Rac is RAC1; RAC2; RAC3; RHOG, or a mutant thereof.

    [1012] Embodiment 127. The compound or pharmaceutically acceptable derivative of any one of embodiments 1-126, wherein the pharmaceutically acceptable derivative of the compound is a pharmaceutically acceptable salt of said compound.

    [1013] Embodiment 128. A method of inhibiting the function of one or more members of the Ras superfamily, comprising administering to a subject a compound which inhibits the one or more members of the Ras superfamily with an IC.sub.50 value of less than 10 μM, wherein the compound is the compound or pharmaceutically acceptable derivative of any one of embodiments 1-126 or the compound is the compound or pharmaceutically acceptable salt of embodiment 127.

    [1014] Embodiment 129. The method of embodiment 128, wherein one or more members of the Ras superfamily is Ras.

    [1015] Embodiment 130. The method of embodiment 128, wherein one or more members of the Ras superfamily is Rho.

    [1016] Embodiment 131. The method of embodiment 128, wherein one or more members of the Ras superfamily is Rac.

    [1017] Embodiment 132. The method of embodiment 128, wherein the Ras is DIRAS1; DIRAS2; DIRAS3; ERAS; GEM; HRAS; KRAS; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS; or RRAS2.

    [1018] Embodiment 133. The method of embodiment 132, wherein the Ras is HRAS, KRAS, NRAS or a mutant thereof

    [1019] Embodiment 134. The method of embodiment 132, wherein the Ras is HRAS or a mutant thereof.

    [1020] Embodiment 135. The method of embodiment 132, wherein the Ras is KRAS or a mutant thereof.

    [1021] Embodiment 136. The method of embodiment 132, wherein the Ras is NRAS or a mutant thereof.

    [1022] Embodiment 137. The method of embodiment 130, wherein the Rho is RHOA; RHOB; RHOBTB1; RHOBTB2; RHOBTB3; RHOC; RHOD; RHOF; RHOG; RHOH; RHOJ; RHOQ; RHOU; RHOV; RND1; RND2; RND3; RAC1; RAC2; RAC3; CDC42, or a mutant thereof.

    [1023] Embodiment 138. The method of embodiment 137, wherein the Rho is Rac.

    [1024] Embodiment 139. The method of embodiment 131 or 138, wherein the Rac is RAC1; RAC2; RAC3; RHOG, or a mutant thereof.

    [1025] Embodiment 140. The method of embodiment 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of cancer.

    [1026] Embodiment 141. The method of any of embodiments 129 or 132-136, wherein the inhibiting the function of Ras is a treatment, prevention or amelioration of one or more symptoms of cancer.

    [1027] Embodiment 142. The method of any of embodiments 130 or 137-138, wherein the inhibiting the function of Rho is a treatment, prevention or amelioration of one or more symptoms of cancer.

    [1028] Embodiment 143. The method of any of embodiments 131 or 138-139, wherein the inhibiting the function of Rac is a treatment, prevention or amelioration of one or more symptoms of cancer.

    [1029] Embodiment 144. The method of any of embodiments 140-143, wherein the cancer is a solid tumor.

    [1030] Embodiment 145. The method of embodiment 144, wherein the solid tumor is hepatocellular carcinoma, prostate cancer, pancreatic cancer, lung cancer, breast cancer, ovarian cancer, colon cancer, small intestine cancer, biliary tract cancer, endometrium cancer, skin cancer (melanoma), cervix cancer, urinary tract cancer, or glioblastoma.

    [1031] Embodiment 146. The method of embodiment 145, wherein the solid tumor is pancreatic cancer.

    [1032] Embodiment 147. The method of embodiment 145, wherein the solid tumor is colon cancer.

    [1033] Embodiment 148. The method of embodiment 145, wherein the solid tumor is small intestine cancer.

    [1034] Embodiment 149. The method of embodiment 145, wherein the solid tumor is biliary tract cancer.

    [1035] Embodiment 150. The method of embodiment 145, wherein the solid tumor is endometrium cancer.

    [1036] Embodiment 151. The method of embodiment 145, wherein the solid tumor is lung cancer.

    [1037] Embodiment 152. The method of embodiment 145, wherein the solid tumor is breast cancer.

    [1038] Embodiment 153. The method of embodiment 145, wherein the solid tumor is skin cancer.

    [1039] Embodiment 154. The method of embodiment 145, wherein the solid tumor is cervix cancer.

    [1040] Embodiment 155. The method of embodiment 145, wherein the solid tumor is urinary tract cancer.

    [1041] Embodiment 156. The method of any of embodiments 140-143, wherein the cancer is a blood borne tumor.

    [1042] Embodiment 157. The method of embodiment 156, wherein the blood borne tumor is a leukemia.

    [1043] Embodiment 158. The method of embodiment 156, wherein the blood borne tumor is chronic lymphocytic leukemia (CLL), chronic myelocytic leukemia (CML), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), or acute myeloblastic leukemia (AML).

    [1044] Embodiment 159. The method of any one of embodiments 156-158, wherein the blood borne tumor is metastatic.

    [1045] Embodiment 160. The method of embodiment 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of an inflammatory disease.

    [1046] Embodiment 161. The method of any of embodiments 129 or 132-136, wherein inhibiting the function of Ras is a treatment, prevention or amelioration of one or more symptoms of an inflammatory disease.

    [1047] Embodiment 162. The method of any of embodiments 130 or 137-138, wherein the inhibiting the function of Rho is a treatment, prevention or amelioration of one or more symptoms of inflammatory disease.

    [1048] Embodiment 163. The method of any of embodiments 131 or 138-139, wherein the inhibiting the function of Rac is a treatment, prevention or amelioration of one or more symptoms of inflammatory disease.

    [1049] Embodiment 164. The method of any of embodiments 160-163, wherein the inflammatory disease is gastritis, schistosomiasis, cholangitis, chronic cholecystitis, pelvic inflammatory disease, chronic cervicitis, osteomyelitis, inflammatory bowel disease, reflux esophagitis, Barrett's esophagus, bladder inflammation (cystitis), asbestosis, silicosis, gingivitis, lichen planus, pancreatitis, protease mutation, lichen sclerosis, slaladenitis, bronchitis, Sjogren syndrome or Hashimoto's thyroiditis.

    [1050] Embodiment 165. The method of any of embodiments 160-163, wherein the inflammatory disease is Alzheimer's disease (AD), ankylosing spondylitis, arthritis (osteoarthritis, rheumatoid arthritis (RA), psoriatic arthritis), asthma, atherosclerosis, Crohn's disease, colitis, dermatitis, diverticulitis, fibromyalgia, hepatitis, irritable bowel syndrome (IBS), systemic lupus, erythematous (SLE), nephritis, Parkinson's disease, ulcerative colitis.

    [1051] Embodiment 166. The method of embodiment 165, wherein the inflammatory disease is Alzheimer's disease (AD).

    [1052] Embodiment 167. The method of embodiment 165, wherein the inflammatory disease is ankylosing spondylitis.

    [1053] Embodiment 168. The method of embodiment 165, wherein the inflammatory disease is arthritis (osteoarthritis, rheumatoid arthritis (RA), psoriatic arthritis).

    [1054] Embodiment 169. The method of embodiment 165, wherein the inflammatory disease is asthma.

    [1055] Embodiment 170. The method of embodiment 165, wherein the inflammatory disease is atherosclerosis.

    [1056] Embodiment 171. The method of embodiment 165, wherein the inflammatory disease is Crohn's disease.

    [1057] Embodiment 172. The method of embodiment 165, wherein the inflammatory disease is colitis.

    [1058] Embodiment 173. The method of embodiment 165, wherein the inflammatory disease is dermatitis.

    [1059] Embodiment 174. The method of embodiment 165, wherein the inflammatory disease is diverticulitis.

    [1060] Embodiment 175. The method of embodiment 165, wherein the inflammatory disease is fibromyalgia.

    [1061] Embodiment 176. The method of embodiment 165, wherein the inflammatory disease is hepatitis.

    [1062] Embodiment 177. The method of embodiment 165, wherein the inflammatory disease is irritable bowel syndrome (IBS).

    [1063] Embodiment 178. The method of embodiment 165, wherein the inflammatory disease is systemic lupus.

    [1064] Embodiment 179. The method of embodiment 165, wherein the inflammatory disease is erythematous (SLE).

    [1065] Embodiment 180. The method of embodiment 165, wherein the inflammatory disease is nephritis.

    [1066] Embodiment 181. The method of embodiment 165, wherein the inflammatory disease is Parkinson's disease.

    [1067] Embodiment 182. The method of embodiment 165, wherein the inflammatory disease is ulcerative colitis.

    [1068] Embodiment 183. The method of embodiment 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of a rasopathy.

    [1069] Embodiment 184. The method of any of embodiments 129 or 132-136, wherein the inhibiting the function of Ras is a treatment for a rasopathy.

    [1070] Embodiment 185. The method of any of embodiments 130 or 137-138, wherein the inhibiting the function of Rho is a treatment for a rasopathy.

    [1071] Embodiment 186. The method of any of embodiments 131 or 138-139, wherein the inhibiting the function of Rac is a treatment for a rasopathy.

    [1072] Embodiment 187. The method of any of embodiments 183-186, wherein the rasopathy is neurofibromatosis type 1, Noonan's syndrome or Costello syndrome.

    [1073] Embodiment 188. The method of any of embodiments 129 or 132-136, wherein the inhibiting the function of Ras is a treatment for Ras-associated autoimmune leukoproliferative disorder.

    [1074] Embodiment 189. The method of embodiment 128, wherein the inhibiting the function of one or more members of the Ras superfamily is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    [1075] Embodiment 190. The method of any of embodiments 129 or 132-136, wherein the inhibiting the function of Ras is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    [1076] Embodiment 191. The method of any of embodiments 130 or 137-138, wherein the inhibiting the function of Rho is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    [1077] Embodiment 192. The method of any of embodiments 130 or 137-138, wherein the inhibiting the function of Rac is a treatment, prevention or amelioration of one or more symptoms of a fibrotic disease.

    [1078] Embodiment 193. The method of any one of embodiments 140, 160, 183, or 189, wherein one or more members of the Ras superfamily is Ras.

    [1079] Embodiment 194. The method of any one of embodiments 140, 160, 183, or 189, wherein one or more members of the Ras superfamily is Rho.

    [1080] Embodiment 195. The method of any one of embodiments 140, 160, 183, or 189, wherein one or more members of the Ras superfamily is Rac.

    [1081] Embodiment 196. A pharmaceutical composition, comprising the compound or pharmaceutically acceptable derivative of any one of embodiments 1-126, and a pharmaceutically acceptable carrier.

    [1082] Embodiment 197. The pharmaceutical composition of embodiment 196, wherein the pharmaceutical composition comprises a therapeutic amount of said compound or pharmaceutically acceptable derivative thereof.

    [1083] Embodiment 198. A pharmaceutical composition, comprising the compound or pharmaceutically acceptable salt of embodiment 127, and a pharmaceutically acceptable carrier.

    [1084] Embodiment 199. The pharmaceutical composition of embodiment 198, wherein the pharmaceutical composition comprises a therapeutic amount of said compound or pharmaceutically acceptable salt thereof.

    [1085] Embodiment 200. A method of inhibiting the function of one or more members of the Ras superfamily, comprising administering to a subject the pharmaceutical composition of any one of embodiments 196-199.

    [1086] Embodiment 201. A method of inhibiting the function of one or more members of the Ras superfamily, comprising administering to a subject the compound or pharmaceutically acceptable derivative of any one of embodiments 1-126.

    [1087] This disclosure is not to be limited in scope by the embodiments disclosed in the examples which are intended as single illustrations of individual aspects, and any methods which are functionally equivalent are within the scope of this disclosure. Indeed, various modifications in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.

    [1088] Various references such as patents, patent applications, and publications are cited herein, the disclosures of which are hereby incorporated by reference herein in their entireties.