FOLEY KEVIN (US)
YING CHENGHAO (CN)
WANG ZHIYONG (CN)
YIN WEI (CN)
WANG YAYA (CN)
DAI YAN (CN)
PRINCE THOMAS (CN)
WANG GUOQIANG (US)
YING WEIWEN (US)
FOLEY KEVIN P (US)
YING CHENGHAO (CN)
WANG ZHIYONG (CN)
YIN WEI (CN)
WANG YAYA (CN)
DAI YAN (CN)
PRINCE THOMAS (CN)
WANG GUOQIANG (US)
WO2023098426A1 | 2023-06-08 | |||
WO2022187528A1 | 2022-09-09 | |||
WO2023081476A1 | 2023-05-11 | |||
WO2022235866A1 | 2022-11-10 | |||
WO2021041671A1 | 2021-03-04 | |||
WO2022015375A1 | 2022-01-20 | |||
WO2022066646A1 | 2022-03-31 | |||
WO2017201161A1 | 2017-11-23 |
MCCOY ET AL., NATURE, vol. 302, 1983, pages 79 - 8
COXDER, SMALL GTPASES, vol. 1, 2010, pages 2 - 27
KERK ET AL., NAT REV CANCER, vol. 21, 2021, pages 510 - 525
PYLAYEVA-GUPTA ET AL., NAT REV CANCER, vol. 11, 2011, pages 761 - 774
ZHOU ET AL., PATHOL ONCOL RES, vol. 26, 2020, pages 2835 - 2837
AREDO ET AL., LUNG CANCER, vol. 133, 2019, pages 144 - 150
OLMEDILLAS-LOPEZ ET AL., WORLD J GASTROENTEROL, vol. 23, no. 39, 2017, pages 7087 - 709
MIGLIO ET AL., PATHOL RES PRACT, vol. 210, 2014, pages 307 - 11
GOU ET AL., BR J CANCER, vol. 22, 2020, pages 857 - 867
MCCORMICK F, BIOCHEM J, vol. 476, 2019, pages 356 - 74
OSTREM ET AL., NATURE, vol. 503, 2013, pages 548 - 51
CLAIMS g of Claims: 1. A compound of the structural formula I*: (I*); or a pharmaceutically acceptable salt thereof, wherein L is selected from -C(=O), -S(=O), and -S(O)2; R0 is a chemical entity which binds to KRASG12D; and R00 is selected from hydrogen, F, Cl, and CF3. 2. The compound of Claim 1, wherein the compound is of the structural formula I: or a pharmaceutically acceptable salt thereof. 3. The compound of Claim 1 or 2, wherein the compound is of the structural formula Ia or Ib: or a pharmaceutically acceptable salt thereof, wherein R1 is selected from (C1-C4)alkylO(C1-C4)alkyl, -(C1-C4)alkylNH(C1-C4)alkyl, -(C1- C4)alkylN[(C1-C4)alkyl]2, heterocyclyl, and cycloalkyl, wherein said heterocyclyl and cycloalkyl are each optionally and independently substituted; R2 is selected from hydrogen, (C1-C4)alkyl, (C1-C4)haloalkyl, (C1-C4)alkoxy, (C1- C4)haloalkoxy, (C1-C4)alkynyl, (C1-C4)alkenyl, halo, (C3-C6)cycloalkyl, -O(C3- C6)cycloalkyl, cyano, NH2, -NH(C1-C4)alkyl, -N[(C1-C4)alkyl]2, -P(O)[(C1-C4)alkyl]2, and - S(C1-C4)alkyl; R3 is selected from a1 is N or CHZ1; Z1 is selected from hydrogen, halo, (C1-C4)alkyl, (C2-C4)alkenyl, cyano, cyano(C1- C4)alkyl, -S[halo(C1-C4)alkyl], and (C3-C6)cycloalkyl; R8, R9, R10, R11, R12, and R13 are each independently selected from hydrogen, (C1- C4)alkyl, (C1-C4)haloalkyl, (C1-C4)cyanoalkyl, (C1-C4)hydroxyalkyl, -(C1-C4)alkylNRaRb, - (C1-C4)alkylC(O)NRaRb, (C1-C4)alkylO(C1-C4)alkyl, -(C1-C4)alkylC(O)ORa, -(C1- C4)alkylNRaC(O)ORb, -(C1-C4)alkylC(O)Ra, -(C1-C4)alkylheterocyclyl, -(C1-C4)alkylaryl, - (C1-C4)alkylheteroaryl, (C2-C4)alkenyl, (C2-C4)haloalkenyl, (C2-C4)cyanoalkenyl, (C2- C4)hydroxyalkenyl, -(C2-C4)alkenylNRaRb, (C2-C4)alkynyl, (C2-C4)haloalkynyl, (C2- C4)cyanoalkynyl, (C2-C4)hydroxyalkynyl, -(C2-C4)alkynylNRaRb, (C1-C4)alkoxy, (C1- C4)haloalkoxy, halo, cyano, oxo, hydroxy, -S(C1-C4)alkyl, -S(C1-C4)haloalkyl, -NRaRb, - NRaC(O)Rb, -C(O)Ra, -C(O)ORa, -SO2Ra, -S(O)Ra, -SO2NRaRb, -NRaSO2Rb, (C3- C6)cycloalkyl, 5- or 6-membered heteroaryl, and 4- to 6-membered heterocyclyl, wherein said heterocyclyl, aryl, and heteroaryl of -(C1-C4)alkylheterocyclyl, -(C1-C4)alkylaryl, and - (C1-C4)alkylheteroaryl, and said (C3-C6)cycloalkyl, 5- or 6-membered heteroaryl, and 4- to 6-membered heterocyclyl are each optionally and independently substituted with 1 to 3 groups selected from Rc; m is 0, 1, or 2; Ra and Rb are each independently selected from hydrogen, (C1-C4)alkyl, and (C1- C4)haloalkyl, or Ra and Rb, when on the same nitrogen atom, may be taken together to form a heterocyclyl; and Rc is selected from halo, (C1-C4)alkyl, (C1-C4)haloalkyl, (C1-C4)alkoxy, (C1- C4)haloalkoxy, cyano, hydroxyl, oxo, -C(O)ORa, -C(O)Ra, -SO2Ra, -S(O)Ra, -SO2NRaRb, - NRaC(O)Rb, -NRaSO2Rb, -NRaRb, and NO2. 4. The compound of any one of Claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein Z is selected from hydrogen, methyl, ethyl, isopropyl, cyano, -SCF3, cyanoethyl, cyclopropyl, and 2-methylpropenyl. 5. The compound of any one of Claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from an optionally substituted heterocyclyl and an optionally substituted cycloalkyl. 6. The compound of any one of Claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from an optionally substituted fused bicyclic heterocyclyl and an optionally substituted (C3-C6)cycloalkyl. 7. The compound of any one of Claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from R4, R6, and R7 are each independently selected from hydrogen, (C1-C4)alkyl, (C1- C4)haloalkyl, (C1-C4)cyanoalkyl, (C1-C4)hydroxyalkyl, -(C1-C4)alkylNRaRb, -(C1- C4)alkylC(O)NRaRb, (C1-C4)alkylO(C1-C4)alkyl, -(C1-C4)alkylC(O)ORa, -(C1- C4)alkylNRaC(O)ORb, -(C1-C4)alkylC(O)Ra, -(C1-C4)alkylheterocyclyl, -(C1-C4)alkylaryl, - (C1-C4)alkylheteroaryl, (C2-C4)alkenyl, (C2-C4)haloalkenyl, (C2-C4)cyanoalkenyl, (C2- C4)hydroxyalkenyl, -(C2-C4)alkenylNRaRb, (C2-C4)alkynyl, (C2-C4)haloalkynyl, (C2- C4)cyanoalkynyl, (C2-C4)hydroxyalkynyl, -(C2-C4)alkynylNRaRb, (C1-C4)alkoxy, (C1- C4)haloalkoxy, halo, cyano, oxo, hydroxy, -S(C1-C4)alkyl, -S(C1-C4)haloalkyl, -NRaRb, - NRaC(O)Rb, -C(O)Ra, -C(O)ORa, -SO2Ra, -S(O)Ra, -SO2NRaRb, -NRaSO2Rb, (C3- C6)cycloalkyl, 5- or 6-membered heteroaryl, and 4- to 6-membered heterocyclyl, wherein said heterocyclyl, aryl, and heteroaryl of -(C1-C4)alkylheterocyclyl, -(C1-C4)alkylaryl, and - (C1-C4)alkylheteroaryl, and said (C3-C6)cycloalkyl, 5- or 6-membered heteroaryl, and 4- to 6-membered heterocyclyl are each optionally and independently substituted with 1 to 3 groups selected from Rc; R5 is (C1-C4)alkyl or halo; and n, x and z are each independently 0, 1, or 2. 8. The compound of any one of Claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from , , . 9. The compound of any one of Claims 1 to 8, wherein the compound is of the structural formula Ia’: or a pharmaceutically acceptable salt thereof. 10. The compound of any one of Claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein a1 is N. 11. The compound of any one of Claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R00 is F. 12. The compound of any one of Claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein . 13. The compound of any one of Claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein R8, R9, R10, and R11 are each independently selected from hydrogen, halo, hydroxyl, and (C2-C4)alkynyl. 14. The compound of any one of Claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R8, R9, R10, and R11 are each hydrogen; or R9, R10 and R11 are each hydrogen and R8 is selected from halo, hydroxyl, and (C2- C4)alkynyl; or R10 and R11 are each hydrogen and R8 and R9 are each independently selected from halo, hydroxyl, and (C2-C4)alkynyl; or R11 is hydrogen and R8, R9, and R10 are each independently selected from halo, hydroxyl, and (C2-C4)alkynyl. 15. The compound of any one of Claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein R8, R9, R10 and R11 are each hydrogen; or R9, R10 and R11 are each hydrogen and R8 is (C2-C4)alkynyl; or R10 and R11 are each hydrogen, R8 is (C2-C4)alkynyl, and R9 is halo; or R11 is hydrogen, R8 is (C2-C4)alkynyl, R9 is halo, and R10 is hydroxyl. 16. The compound of any one of Claims 1 to 15, or a pharmaceutically acceptable salt thereof, wherein R3 is or a pharmaceutically acceptable salt thereof. 17. The compound of any one of Claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein R3 is or a pharmaceutically acceptable salt thereof. 18. The compound of any one of Claims 1 to 11, or a pharmaceutically acceptable salt 19. The compound of any one of Claims 1 to 18, or a pharmaceutically acceptable salt thereof, wherein m is 0 or 1. 20. The compound of any one of Claims 1 to 19, or a pharmaceutically acceptable salt thereof, wherein m is 1. 21. The compound of any one of Claims 1 to 20, or a pharmaceutically acceptable salt thereof, wherein R5 is halo. 22. The compound of any one of Claims 1 to 21, or a pharmaceutically acceptable salt thereof, wherein z is 0 or 2. 23. The compound of any one of Claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein z is 0. 24. The compound of any one of Claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein . 25. The compound of any one of Claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein R4 is -(C1-C4)alkylheterocyclyl optionally substituted with 1 to 3 groups selected from Rc. 26. The compound of any one of Claims 1 to 25, or a pharmaceutically acceptable salt thereof, wherein R4 is each optionally substituted with 1 to 3 groups selected from Rc. 27. The compound of any one of Claims 1 to 26, or a pharmaceutically acceptable salt thereof, wherein Rc is selected from (C1-C4)alkyl, (C1-C4)haloalkyl, and halo. 28. The compound of any one of Claims 1 to 27, or a pharmaceutically acceptable salt thereof, wherein Rc is selected from (C1-C4)haloalkyl and halo. 29. The compound of any one of Claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R1 is selected from . 30. The compound of any one of Claims 1 to 29, or a pharmaceutically acceptable salt thereof, wherein R2 is selected from hydrogen, (C1-C4)alkyl, (C1-C4)haloalkyl, (C1- C4)alkoxy, (C1-C4)haloalkoxy, (C1-C4)alkynyl, (C1-C4)alkenyl, halo, (C3-C6)cycloalkyl, cyano, NH2, -NH(C1-C4)alkyl, -N[(C1-C4)alkyl]2, -P(O)[(C1-C4)alkyl]2, and -S(C1-C4)alkyl. 31. The compound of any one of Claims 1 to 30, or a pharmaceutically acceptable salt thereof, wherein R2 is selected from hydrogen and (C1-C4)alkoxy. 32. The compound of any one of Claims 1 to 29, or a pharmaceutically acceptable salt thereof, wherein R2 is selected from hydrogen, chloro, cyano, methoxy, ethoxy, CF3, and 33. The compound of Claim 1, wherein the compound is selected from or a pharmaceutically acceptable salt thereof. 34. A pharmaceutical composition comprising a compound of any one of Claims 1 to 33, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier. 35. A method for treating cancer in a subject comprising administering to the subject and effective amount of a compound of any one of Claims 1 to 33, or a pharmaceutically acceptable salt thereof, or the pharmaceutically acceptable composition of Claim 34. |
variables are as described above for formula Ia, Ia’, or Ib or any one of the fourth or sixth to fourteenth embodiments. As part of a sixteenth embodiment, R 4 in the compounds and pharmaceutically acceptable salts described herein is -(C1-C4)alkylheterocyclyl optionally substituted with 1 to 3 groups selected from R c , wherein the remaining variables are as described above for formula Ia, Ia’, or Ib or any one of the fourth or sixth to fifteenth embodiments. Alternatively, as part of a sixteenth embodiment, R 4 is each optionally substituted with 1 to 3 groups selected from R c , wherein the remaining variables are as described above for formula Ia, Ia’, or Ib or any one of the fourth or sixth to fifteeenth embodiments. As part of a seventeenth embodiment, R c in the compound of structural formula Ia, Ia’, or Ib, or a pharmaceutically acceptable salt thereof, is selected from (C 1 -C 4 )alkyl, (C 1 - C4)haloalkyl, and halo, wherein the remaining variables are as described above for formula Ia, Ia’, or Ib or any one of the fourth or sixth to sixteenth embodiments. Alternatively, as part of a seventeenth embodiment, R c in the compound of structural formula Ia, Ia’, or Ib, or a pharmaceutically acceptable salt thereof, is selected from (C1-C4)haloalkyl and halo, wherein the remaining variables are as described above for formula Ia, Ia’, or Ib or any one of the fourth or sixth to sixteenth embodiments. As part of an eighteenth embodiment, R 2 in the compound of structural formula Ia or Ia’ is selected from hydrogen, (C1-C4)alkyl, (C1-C4)haloalkyl, (C1-C4)alkoxy, (C1- C 4 )haloalkoxy, (C 1 -C 4 )alkynyl, (C 1 -C 4 )alkenyl, halo, (C 3 -C 6 )cycloalkyl, cyano, NH 2 , - NH(C 1 -C 4 )alkyl, -N[(C 1 -C 4 )alkyl] 2 , -P(O)[(C 1 -C 4 )alkyl] 2 , and -S(C 1 -C 4 )alkyl, wherein the remaining variables are as described above for formula Ia or Ia’ or any one of the fourth or sixth to seventeenth embodiments. Alternatively, as part of an eighteenth embodiment, R 2 in the compound of structural formula Ia or Ia’, or a pharmaceutically acceptable salt thereof, is selected from hydrogen and (C1-C4)alkoxy, wherein the remaining variables are as described above for formula Ia or Ia’ or any one of the fourth or sixth to seventeenth embodiments. In another alternative, as part of an eighteenth embodiment, R 2 in the compound of structural formula Ia or Ia’, or a pharmaceutically acceptable salt thereof, is selected from hydrogen, chloro, cyano, methoxy, ethoxy, CF 3 , OCF 3 , and , wherein the remaining variables are as described above for formula Ia or Ia’ or any one of the fourth or sixth to seventeenth embodiments. Additional compounds are further disclosed in the Exemplification and are included in the present disclosure. Pharmaceutically acceptable salts thereof as well as the neutral forms are included. 4. Uses, Formulation and Administration Compounds and compositions described herein are generally useful as anticancer therapies. In one aspect, the disclosed compounds and compositions behave as inhibitors of KRAS(G12D). Their mechanisms of action include, but are not limited to, inhibiting KRAS(G12D) and thereby impeding down-stream signals that may result in inhibition of cancer cell growth and/or induction of cancer cell death or other KRAS or KRAS(G12D) functions. In one aspect, the disclosed compounds effectuate the inhibition of KRAS(G12D). Thus, provided herein are methods of treating conditions which are responsive to the inhibition of KRAS(G12D) comprising administering to a subject in need thereof, a therapeutically effective amount of one or more compounds or compositions described herein. Also provided is the use of one or more compounds or compositions described herein in the manufacture of a medicament for treating conditions which are responsive to the inhibition of KRAS(G12D). Further provided is the use of a compound or composition described herein for treating conditions which are responsive to the inhibition of KRAS(G12D). In one aspect, the condition treated by the present compounds and compositions is a cancer. The terms "cancer" or "tumor" are well known in the art and refer to the presence, e.g., in a subject, of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, decreased cell death/apoptosis, and certain characteristic morphological features. Cancer cells are often in the form of a solid tumor. However, cancer also includes non-solid tumors, e.g., blood tumors, e.g., leukemia, wherein the cancer cells are derived from bone marrow. As used herein, the term "cancer" includes pre-malignant as well as malignant cancers. Cancers include, but are not limited to, acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia (monocytic, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocytic and promyelocytic), acute T-cell leukemia, basal cell carcinoma, bile duct carcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myelocytic (granulocytic) leukemia, chronic myelogenous leukemia, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, dysproliferative changes (dysplasias and metaplasias), embryonal carcinoma, endometrial cancer, endotheliosarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen-receptor positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell testicular cancer, glioma, heavy chain disease, hemangioblastoma, hepatoma, hepatocellular cancer, hormone insensitive prostate cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphagioendotheliosarcoma, lymphangiosarcoma, lymphoblastic leukemia, lymphoma (Hodgkin and non-Hodgkin), malignancies and hyperproliferative disorders of the bladder, breast, colon, lung, ovaries, pancreas, prostate, skin, and uterus, lymphoid malignancies of T-cell or B-cell origin, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myelogenous leukemia, myeloma, myxosarcoma, neuroblastoma, non-small cell lung cancer, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinomas, papillary carcinoma, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung carcinoma, solid tumors (carcinomas and sarcomas), small cell lung cancer, stomach cancer, squamous cell carcinoma, synovioma, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular tumors, uterine cancer, and Wilms' tumor. Other cancers include primary cancer, metastatic cancer, oropharyngeal cancer, hypopharyngeal cancer, liver cancer, gall bladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, kidney cancer, urothelium cancer, female genital tract cancer, uterine cancer, gestational trophoblastic disease, male genital tract cancer, seminal vesicle cancer, testicular cancer, germ cell tumors, endocrine gland tumors, thyroid cancer, adrenal cancer, pituitary gland cancer, hemangioma, sarcoma arising from bone and soft tissues, Kaposi's sarcoma, nerve cancer, ocular cancer, meningial cancer, glioblastomas, neuromas, neuroblastomas, Schwannomas, solid tumors arising from hematopoietic malignancies such as leukemias, metastatic melanoma, recurrent or persistent ovarian epithelial cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumors, colorectal cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small-cell lung cancer, malignant glioma, epithelial ovarian cancer, primary peritoneal serous cancer, metastatic liver cancer, neuroendocrine carcinoma, refractory malignancy, triple negative breast cancer, HER2- amplified breast cancer, nasopharageal cancer, oral cancer, biliary tract, hepatocellular carcinoma, squamous cell carcinomas of the head and neck (SCCHN), non-medullary thyroid carcinoma, recurrent glioblastoma multiforme, neurofibromatosis type 1, CNS cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral/lentiginous melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumor, triple negative breast cancer, colorectal cancer, sarcoma, melanoma, renal carcinoma, endometrial cancer, thyroid cancer, rhabdomysarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumor, mantle cell lymphoma, and refractory malignancy. "Solid tumor," as used herein, is understood as any pathogenic tumor that can be palpated or detected using imaging methods as an abnormal growth having three dimensions. A solid tumor is differentiated from a blood tumor such as leukemia. However, cells of a blood tumor are derived from bone marrow; therefore, the tissue producing the cancer cells is a solid tissue that can be hypoxic. "Tumor tissue” or “tumorous tissue" are understood as cells, extracellular matrix, and other naturally occurring components associated with the solid tumor. A specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound described herein in the composition will also depend upon the particular compound in the composition. EXEMPLIFICATION Chemical Synthesis The representative examples that follow are intended to help illustrate the present disclosure, and are not intended to, nor should they be construed to, limit the scope of the invention. General starting materials used were obtained from commercial sources or prepared in other examples, unless otherwise noted. Preparation of Compounds The compounds claimed herein were prepared following the procedures outlined in the following protocols. Intermediate 2: 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine.2 batches: To a mixture of Intermediate 1 (50.0 g, 232 mmol, 1.00 eq) in Tol. (150 mL) was added POCl3 (178 g, 1.16 mol, 108 mL, 5.00 eq) at 25 °C. Then DIEA (65.9 g, 510 mmol, 88.9 mL, 2.20 eq) was added into the mixture blow 40 °C. The mixture was stirred at 110 °C for 12 hrs. LC-MS showed desired MS was detected. The reaction mixture was distilled under reduced pressure to remove POCl3 at 90 °C. The residue was poured into Sat.NaHCO3 slowly (keep pH = 8). During this period, yellow precipitate was formed. It was collected by filtration and washed by H 2 O. The solid was used for next step directly. Intermediate 2 (101 g, 400 mmol, 86.2% yield) was obtained as brown solid. HNMR (DMSO-d6, 400 MHz): δ 8.92-8.86 (m, 1H). LC-MS: m/z 253.9 [M+H] + . Intermediate 3: tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate.2 batches: To a mixture of Intermediate 2 (48.5 g, 192 mmol, 1.00 eq) in DCM (485 mL) was added a solution of compound 2a (38.7 g, 183 mmol, 0.950 eq) in DCM (120 mL). Then DIEA (49.7 g, 384 mmol, 66.9 mL, 2.00 eq) was added into the mixture at -40 °C and stirred at -40 °C for 0.5 hr under N2. LCMS showed compound 2 was consumed, and desired MS was detected. The mixture was quenched by HCl (0.5 M) and the pH of the aqueous phase was acidified to 6~7, then separated, the organic layer was dried over Na2SO4. The residue was purified by column chromatography (SiO2, TLC:Petroleum ether : Ethyl acetate = 3: 1, Rf = 0.4, Petroleum ether : Ethyl acetate = 10: 1 to 1: 1, Rf = 0.4). Intermediate 3(120 g, 280 mmol, 72.9% yield) was obtained as white solid. HNMR (DMSO-d 6 , 400 MHz): δ 9.13-8.98 (m, 1H), 4.67-4.36 (m, 2H), 4.35-4.21 (m, 2H), 3.87-3.50 (m, 2H), 1.85-1.71 (m, 2H), 1.66-1.56 (m, 2H), 1.46 (s, 9H). LC-MS: m/z 428.0[M+H] + . Intermediate 4: tert-butyl 3-(7-chloro-8-fluoro-2-((1- (hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4- yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate. To a stirred mixture of [1- (hydroxymethyl)cyclopropyl]methanol (14.31 g, 140 mmol, 3 eq) and t-BuONa (13.46 g, 140 mmol, 3 eq) in THF was added tert-butyl 3-{2,7-dichloro-8-fluoropyrido[4,3- d]pyrimidin-4-yl}-3,8-diazabicyclo[3.2.1]octane-8-carboxylat e (20 g, 46.697 mmol, 1 eq) in portions at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 1h under nitrogen atmosphere. The reaction was quenched with Water at 0°C. The resulting mixture was extracted with CH2Cl2 (3 x 10mL). The combined organic layers were washed with sat. NaCl aq. (2x5 mL), dried over anhydrous Na 2 SO 4 . The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford intermediate 4 (14.5 g, 62.86%) as a white solid. LCMS (ES, m/z): 494 [M+H] + Intermediate 5: tert-butyl 3-(8-fluoro-7-(7-fluoro-2-(methoxymethoxy)-8-((triisopropyls ilyl)ethy nyl)naphthalen-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methox y)pyrido[4,3-d]pyrimi din-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate. A solution of intermediate 4 (3 g, 6.073 mmol, 1.0 eq) and ((3-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-d ioxa borolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (2.63 g, 9.110 mmol, 1.5 eq) in 1,4- dioxane(40 mL) was treated with Pd(PPh 3 ) 4 (1.40 g, 1.215 mmol, 0.2 eq) and K 3 PO 4 (3.87 g , 18.219 mmol, 3.0 eq) under nitrogen atmosphere. The resulting mixture was stirred for ove rnight at 80°C under nitrogen atmosphere. The reaction was quenched with Water at room t emperature. The resulting mixture was extracted with CH 2 Cl 2 (3 x 10mL). The combined or ganic layers were washed with sat. NaCl aq. (2x5 mL), dried over anhydrous Na2SO4. The r esulting mixture was concentrated under reduced pressure. The residue was purified by silic a gel column chromatography, eluted with CH 2 Cl 2 / MeOH (10:1) to afford intermediate 5 ( 2.5 g, 66.38%) as a yellow solid. LCMS (ES, m/z): 620 [M+H] + Intermediate 6: tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1- formylcyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate. To a stirred solution of intermediate 5 (200 mg, 0.323 mmol, 1 eq) in DCM was added Dess-Martin (410.38 mg, 0.969 mmol, 3 eq) in portions at room temperature. The resulting mixture was stirred for 2h at room temperature. The reaction was quenched with sat. NaHCO3 (aq.) at 0°C.The resulting mixture was extracted with CH2Cl2 (3 x 10mL). The combined organic layers were washed with sat. NaCl (aq.) (3x5 mL), dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The crude product was used in the next step directly without further purification. LCMS (ES, m/z): 618 [M+H] + Intermediate 7: tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropyls ilyl)ethy nyl)naphthalen-1-yl)-2-((1-((4-fluoropiperidin-1-yl)methyl)c yclopropyl)methoxy)pyrid o[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carb oxylate. To a stirred solutio n of intermediate 6 (100 mg, 0.162 mmol, 1 eq) and 4-fluoropiperidine hydrochloride (45.17 mg, 0.324 mmol, 2 eq) in DMF were added STAB (102.87 mg, 0.486 mmol, 3 eq) in portio ns at room temperature. The resulting mixture was stirred for 16 h at room temperature. The reaction was quenched with Water at room temperature. The resulting mixture was extracte d with EtOAc (3 x 10mL). The combined organic layers were washed with sat. NaCl aq. (2x 5 mL), dried over anhydrous Na2SO4. After filtration, the filtrate is concentrated under redu ced pressure. The crude product was used in the next step directly without further purificatio n. LCMS (ES, m/z): 605 [M+H] + Intermediate 8: 4-(4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-((4- fluoropiperidin-1- yl)methyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-7-yl)-5 -ethynyl-6- fluoronaphthalen-2-ol. To a stirred solution of intermediate 7 (80 mg, 0.132 mmol, 1.0 eq) in DCM was added TFA (1 mL, 13.463 mmol, 101.84 eq) dropwise at room temperature. The resulting mixture was stirred for 1h at room temperature. The resulting mixture was concentrated under reduced pressure. The crude product was purified by Prep-HPLC with the following conditions (Column, XBridge Prep OBD C18 Column, 30x150 mm, 5µm; mobile phase, Water(10 mmol/L NH4HCO3) and ACN (30% ACN up to 80% in 10 min); Detector, uv 220 nm).This resulted in Intermediate 8 (38.15 mg, 47.69%) as a white solid. 1 H NMR (DMSO-d 6 , 400 MHz): δ 10.15 (s, 1H), 9.03 (s, 1H), 7.97 (dd, J = 9.2, 6.0 Hz, 1H), 7.46 (t, J = 9.2 Hz, 1H), 7.39 (d, J = 2.4 Hz, 1H), 7.17 (d, J = 2.4 Hz, 1H), 4.69 (dt, J = 7.6, 3.6 Hz, 1H), 4.48 (d, J = 12.0 Hz, 1H), 4.34 – 4.22 (m, 3H), 3.93 (d, J = 1.2 Hz, 1H), 3.67 – 3.59 (m, 1H), 3.54 (t, J = 5.6 Hz, 3H), 2.74 (s, 1H), 2.56 (s, 2H), 2.30 (t, J = 7.6 Hz, 4H), 1.88 – 1.76 (m, 2H), 1.66 (s, 6H), 0.64 (q, J = 3.2 Hz, 2H), 0.40 (t, J = 3.2 Hz, 2H). LC-MS: m/z 628.9 [M+H] + . Compound 1 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-((1-((4-fluoropi peridin-1-yl)methyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimid in-4-yl)-3,8-diazabicycl o[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one. To a solution of compound 3 (1.00 eq) in DCM (2 mL) was added Py (5.00 eq) and TFAA (1.50 eq) at 0 °C, then the mixture was stir red at 25 °C for 2 hrs. LC-MS showed Reactant 1 was consumed completely and one main p eak with desired mass was detected. The reaction mixture was quenched by addition H 2 O (5 mL) at 10 °C, and extracted with DCM (5 mL * 2). The combined organic layers were was hed with brine (5 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (FA condition; column: Phenome nex lunaC18150*25mm* 10um;mobile phase: [water(FA)-ACN];B%: 18%-48%,10min ). Compound 1 (20.0 mg, 25.4 µmol, 47.1% yield, 93.4% purity, HCl) was obtained as a yell ow solid. 1 H NMR (DMSO-d6, 400 MHz): δ 10.16 (s, 1H), 9.05 (d, J = 1.2 Hz, 1H), 7.98 (d d, J = 9.2, 6.0 Hz, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.40 (d, J = 2.4 Hz, 1H), 7.18 (t, J = 2.4 Hz, 1H), 4.80 (s, 1H), 4.73 (d, J = 12.4 Hz, 1H), 4.65 (s, 2H), 4.59 – 4.48 (m, 1H), 4.35 – 4.24 ( m, 2H), 3.94 (dd, J = 6.4, 1.2 Hz, 1H), 3.76 (d, J = 12.4 Hz, 1H), 3.68 (d, J = 12.4 Hz, 1H), 2.48 (s, 1H), 2.31-2.29 (m, 4H), 1.90 – 1.75 (m, 6H), 1.65 (s, 2H), 0.65 (d, J = 4.8 Hz, 2H), 0.45 – 0.40 (m, 2H). LC-MS: m/z 724.9 [M+H] + . The following compounds were prepared according the general procedures described above using the appropriate starting materials. Compound 2 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one 1 HNMR (400 MHz, DMSO-d6) δ 11.03-10.79 (s, 1H), 10.24 (s, 1H), 9.12 (s, 1H), 8.00 (dd, J = 5.9, 9.3 Hz, 1H), 7.48 (t, J = 9.0 Hz, 1H), 7.42 (d, J = 2.6 Hz, 1H), 7.20 (s, 1H), 5.72- 5.38 (m, 1H), 4.84-4.57 (m, 6H), 3.92 (dd, J = 3.5, 5.3 Hz, 1H),3.86-3.67 (m, 4H), 3.54 (s, 2H), 2.20-1.84 (m, 10H), LC-MS: m/z 697.3 [M+H] + . Compound 3 1-(3-{2-[(1-{3-azabicyclo[3.1.0]hexan-3-ylmethyl}cyclopropyl )methoxy]-7-(8-eth ynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d ]pyrimidin-4-yl}-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethanone.1H NMR (DMSO-d6, 400 MHz): δ 10.15 (s, 1H), 9.05 (s, 1H), 7.98 (dd, J = 9.2, 6.0 Hz, 1H), 7.47 (t, J = 9.2 Hz, 1H), 7.40 (d, J = 2.4 Hz, 1H), 7.18 (t, J = 2.4 Hz, 1H), 4.80 (s, 1H), 4.72 (d, J = 13.6 Hz, 1H), 4.65 (s, 1H), 4.52 (d, J = 12.8 Hz, 1H), 4.33 – 4.18 (m, 2H), 3.92 (dd, J = 6.8, 1.2 Hz, 1H), 3.76 (d, J = 12.8 Hz, 1H), 3.68 (d, J = 12.8 Hz, 1H), 2.96 (dd, J = 8.4, 6.2 Hz, 2H), 2.46 – 2.30 (m, 2H), 2.23 (d, J = 8.4 Hz, 2H), 2.05 – 1.88 (m, 4H), 1.32 (dd, J = 9.6, 5.2 Hz, 2H), 0.59 (s, 2H), 0.55 – 0.50 (m, 1H), 0.40 (s, 2H), 0.25 (p, J = 3.6 Hz, 1H). LC-MS: m/z 705.0 [M+H]+. Compound 4 1-(3-(2-((1-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)cycloprop yl)methoxy)-7-(8- ethynyl-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrim idin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one. 1 H NMR (DMSO-d 6 , 400 MHz): δ 10.14 (s, 1H), 9.04 (s, 1H), 7.89 (dd, J = 8.0, 1.6 Hz, 1H), 7.46 – 7.38 (m, 2H), 7.35 (d, J = 2.4 Hz, 1H), 7.12 (t, J = 2.4 Hz, 1H), 4.80 (s, 1H), 4.73 (dd, J = 12.4, 5.6 Hz, 1H), 4.64 (s, 1H), 4.52 (d, J = 12.8 Hz, 1H), 4.27 – 4.17 (m, 2H), 3.76 (d, J = 12.8 Hz, 1H), 3.68 (d, J = 12.8 Hz, 1H), 3.58 (d, J = 6.4 Hz, 1H), 3.00 – 2.91 (m, 2H), 2.42 -2.35 (m, 2H), 2.23 (d, J = 8.4 Hz, 2H), 2.00 – 1.86 (m, 3H), 1.31 (s, 2H), 0.56-0.52(m, 1H), 0.46 (d, J = 4.0 Hz, 3H), 0.40 (s, 2H), 0.25 (s, 1H). LC-MS: m/z 687.1 [M+H] + . Prophetic Compounds Compound 5 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2R,7aS)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5-methoxyp yrido[4,3-d]pyrimidin-4- yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan- 1-one Compound 6 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-5-methoxy-2-((1- ((4-(trifluoromethyl)piperidin-1-yl)methyl)cyclopropyl)metho xy)pyrido[4,3-d]pyrimidin-4- yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan- 1-one1-((1R,5S)-3-(7-(8- ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-5-methox y-2-((1-((4- (trifluoromethyl)piperidin-1-yl)methyl)cyclopropyl)methoxy)p yrido[4,3-d]pyrimidin-4-yl)- 3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-on e Compound 7 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-5-methoxy-2-((1- (morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin -4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 8 1-(3-(2-((1-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)cycloprop yl)methoxy)-7-(8-ethynyl-7- fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-5-methoxypyrido[4, 3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 9 1-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-f luoro-5-methoxy-4-((1R,5S)- 8-(2,2,2-trifluoroacetyl)-3,8-diazabicyclo[3.2.1]octan-3-yl) pyrido[4,3-d]pyrimidin-2- yl)oxy)methyl)cyclopropyl)methyl)piperidine-4-carbonitrile Compound 10 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-((1-((4- (trifluoromethyl)piperidin-1-yl)methyl)cyclopropyl)methoxy)p yrido[4,3-d]pyrimidin-4-yl)- 3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-on e Compound 11 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-((1-((4- fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)-5-(trifluor omethoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 12 1-((1R,5S)-3-(5-ethoxy-7-(8-ethynyl-7-fluoro-3-hydroxynaphth alen-1-yl)-8-fluoro-2-((1- ((4-fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)pyrido[4 ,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 13 1-((1R,5S)-3-(5-chloro-7-(8-ethynyl-7-fluoro-3-hydroxynaphth alen-1-yl)-8-fluoro-2-((1- ((4-fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)pyrido[4 ,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 14 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-((1-((4- fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)-5-(trifluor omethyl)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 15 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-((1-((4- fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methylpyr ido[4,3-d]pyrimidin-4-yl)- 3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-on e Compound 16 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5-methylpy rido[4,3-d]pyrimidin-4-yl)- 3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-on e Compound 17 1-(3-(5-ethoxy-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-y l)-8-fluoro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 18 1-(3-(5-chloro-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-y l)-8-fluoro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 19 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5-(trifluo romethyl)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 20 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5-(trifluo romethoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 21 1-((1R,5S)-3-(5-ethynyl-7-(8-ethynyl-7-fluoro-3-hydroxynapht halen-1-yl)-8-fluoro-2-((1- ((4-fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)pyrido[4 ,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 22 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-((1-((4- fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)-5-(2-methyl prop-1-en-1-yl)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 23 1-((1R,5S)-3-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-hydroxyn aphthalen-1-yl)-8-fluoro-2- ((1-((4-fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)pyri do[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 24 1-((1R,5S)-3-(5-bromo-7-(8-ethynyl-7-fluoro-3-hydroxynaphtha len-1-yl)-8-fluoro-2-((1- ((4-fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)pyrido[4 ,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 25 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-5,8-difluoro-2-((1-((4- fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)pyrido[4,3-d ]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 26 1-(3-(5-ethynyl-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1- yl)-8-fluoro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 27 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5-(2-methy lprop-1-en-1-yl)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 28 1-(3-(5-cyclopropyl-7-(8-ethynyl-7-fluoro-3-hydroxynaphthale n-1-yl)-8-fluoro-2- (((2S,7aR)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methox y)pyrido[4,3-d]pyrimidin-4- yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan- 1-one Compound 29 1-(3-(5-bromo-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 30 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-5,8-di fluoro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 31 1-((1R,5S)-3-(5-amino-7-(8-ethynyl-7-fluoro-3-hydroxynaphtha len-1-yl)-8-fluoro-2-((1-((4- fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)pyrido[4,3-d ]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 32 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-((1-((4- fluoropiperidin-1-yl)methyl)cyclopropyl)methoxy)-5-(methylam ino)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 33 1-((1R,5S)-3-(5-(dimethylamino)-7-(8-ethynyl-7-fluoro-3-hydr oxynaphthalen-1-yl)-8- fluoro-2-((1-((4-fluoropiperidin-1-yl)methyl)cyclopropyl)met hoxy)pyrido[4,3-d]pyrimidin- 4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroetha n-1-one Compound 34 1-((1R,5S)-3-(5-(dimethylphosphoryl)-7-(8-ethynyl-7-fluoro-3 -hydroxynaphthalen-1-yl)-8- fluoro-2-((1-((4-fluoropiperidin-1-yl)methyl)cyclopropyl)met hoxy)pyrido[4,3-d]pyrimidin- 4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroetha n-1-one Compound 35 1-(3-(5-amino-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one Compound 36 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5-(methyla mino)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one Compound 37 1-(3-(5-(dimethylamino)-7-(8-ethynyl-7-fluoro-3-hydroxynapht halen-1-yl)-8-fluoro-2- (((2S,7aR)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methox y)pyrido[4,3-d]pyrimidin-4- yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan- 1-one Compound 38 1-(3-(5-(dimethylphosphoryl)-7-(8-ethynyl-7-fluoro-3-hydroxy naphthalen-1-yl)-8-fluoro-2- (((2S,7aR)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methox y)pyrido[4,3-d]pyrimidin-4- yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan- 1-one Compound 39 1-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-f luoro-4-((1R,5S)-8-(2,2,2- trifluoroacetyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4, 3-d]pyrimidin-2- yl)oxy)methyl)cyclopropyl)methyl)piperidine-4-carbonitrile Compound 39 1-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-f luoro-4-((1R,5S)-8- (2,2,2-trifluoroacetyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)py rido[4,3-d]pyrimidin-2- yl)oxy)methyl)cyclopropyl)methyl)piperidine-4-carbonitrile. 1 H NMR (DMSO-d 6 , 400 MHz) δ 10.15 (s, 1H), 9.05 (d, J = 1.1 Hz, 1H), 8.00-7.96 (m, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.40 (d, J = 2.6 Hz, 1H), 7.18 (t, J = 2.6 Hz, 1H), 4.80 (s, 1H), 4.73-4.71 (m, 1H), 4.65 (s, 1H), 4.53-4.50 (m, 1H), 4.35 – 4.21 (m, 2H), 3.93 (dd, J = 6.5, 1.1 Hz, 1H), 3.76-3.74 (m, 1H), 3.69-3.67 (m, 1H), 2.82 (s, 1H), 2.55 (s, 2H), 2.32-2.27 (m, 4H), 2.05 – 1.72 (m, 6H), 1.65 (d, J = 8.8 Hz, 2H), 0.65 (s, 2H), 0.43 (s, 2H). LC-MS: m/z 731.90 [M+H] + . Compound 40 1-((1R,5S)-3-(2-((1-((6,6-difluoro-3-azabicyclo[3.1.0]hexan- 3-yl)methyl)cyclo- propyl)methoxy)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1- yl)-8-fluoropyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.15 (s, 1H), 9.05 (d, J = 1.1 Hz, 1H), 7.98 (dd, J = 9.2, 5.9 Hz, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.40 (d, J = 2.6 Hz, 1H), 7.18 (t, J = 2.4 Hz, 1H), 4.80 (s, 1H), 4.73 (dd, J = 13.0, 5.2 Hz, 1H), 4.64 (s, 1H), 4.52 (d, J = 12.9 Hz, 1H), 4.31 – 4.19 (m, 2H), 4.09 (s, 1H), 3.92 (dd, J = 6.4, 1.1 Hz, 1H), 3.76 (d, J = 12.9 Hz, 2H),, 3.01 (dd, J = 9.7, 2.7 Hz, 2H), 2.77 (d, J = 9.7 Hz, 2H), 2.42 (dd, J = 13.2, 10.3 Hz, 2H), 2.28 (dd, J = 13.6, 2.6 Hz, 2H), 1.95 (ddd, J = 30.3, 14.9, 7.2 Hz, 4H), 0.61 (q, J = 3.2 Hz, 2H), 0.43 (t, J = 2.9 Hz, 2H). LC-MS: m/z 740.80 [M+H] + . Compound 41 3-((1-(((7-(8-ethynyl-3-hydroxynaphthalen-1-yl)-8-fluoro-4-( 8-(2,2,2-trifluoro-acetyl)- 3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-y l)oxy)methyl) cyclopropyl)methyl)-3-azabicyclo[3.1.0]hexane-1-carbonitrile . 1 H NMR (DMSO-d 6 , 400 MHz) δ 10.13 (s, 1H), 9.04 (s, 1H), 7.89 (dd, J = 8.0, 1.7 Hz, 1H), 7.48 – 7.37 (m, 2H), 7.35 (d, J = 2.6 Hz, 1H), 7.12 (t, J = 2.4 Hz, 1H), 4.80 (s, 1H), 4.73-4.70 (m, 1H), 4.64 (s, 1H), 4.54-4.51 (m, 1H), 4.24 (d, J = 2.3 Hz, 2H), 3.75-3.66 (m, 2H), 3.57 (dd, J = 6.5, 1.3 Hz, 1H), 3.22 (dd, J = 8.5, 1.5 Hz, 1H), 2.98 (dd, J = 9.3, 3.1 Hz, 1H), 2.47 – 2.34 (m, 4H), 2.18 (qd, J = 4.7, 2.2 Hz, 1H), 2.08 – 1.77 (m, 4H), 1.35 – 1.25 (m, 1H), 1.09 (dt, J = 8.0, 3.8 Hz, 1H), 0.60 (s, 2H), 0.43-0.41 (m, 2H). LC-MS: m/z 711.90 [M+H] + . Compound 42 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-((1-((4-fluoro- piperidin-1-yl)methyl)cyclopropyl)methoxy)-5-methoxypyrido[4 ,3-d]pyrimidin-4-yl)- 3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-on e. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.17 (s, 1H), 8.05-7.90 (m, 1H), 7.55-7.35 (m, 2H), 7.22 (s, 1H), 4.87-4.53 (m, 2H), 4.40-4.08 (m, 3H), 4.01-3.81 (m, 4H), 3.54-3.40 (m, 3H), 3.18-2.91 (m, 2H), 2.40- 2.27 (m, 1H), 2.03-1.61 (m, 8H), 1.55-1.28 (m, 2H), 0.80-0.34 (m, 4H). LC-MS: m/z 755.5 [M+H] + . Compound 43 1-((1R,5S)-3-(7-(8-ethynylnaphthalen-1-yl)-8-fluoro-2-((1-(( 4-fluoropiperidin-1-yl) methyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1] octan-8-yl)-2,2,2-trifluoroethan-1-one. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.07 (d, J = 1.1 Hz, 1H), 8.20 – 8.11 (m, 2H), 7.77 – 7.67 (m, 2H), 7.63 – 7.54 (m, 2H), 4.80 (d, J = 5.9 Hz, 1H), 4.73 (d, J = 13.5 Hz, 1H), 4.66 (s, 2H), 4.53 (d, J = 13.0 Hz, 1H), 4.36 – 4.24 (m, 2H), 3.77 (d, J = 12.8 Hz, 1H), 3.73 – 3.65 (m, 2H), 2.57 (s, 2H), 2.38 – 2.25 (m, 4H), 2.11 – 1.75 (m, 6H), 1.65 (t, J = 9.5 Hz, 2H), 0.66 (q, J = 3.5, 3.0 Hz, 2H), 0.43 (t, J = 2.9 Hz, 2H). LC-MS: m/z 690.90 [M+H] + . Compound 44 5-ethynyl-6-fluoro-4-(8-fluoro-2-((1-((4-fluoropiperidin-1-y l)methyl)cyclopropyl) methoxy)-4-((1R,5S)-8-(2,2,2-trifluoroacetyl)-3,8-diazabicyc lo[3.2.1]octan-3-yl)pyrido [4,3-d]pyrimidin-7-yl)naphthalen-2-yl isobutyrate. 1 H NMR (DMSO-d 6 , 400 MHz) δ 9.07 (d, J = 1.8 Hz, 1H), 8.22 (dd, J = 9.2, 5.8 Hz, 1H), 8.01 (d, J = 2.6 Hz, 1H), 7.66 (t, J = 9.0 Hz, 1H), 7.49 (t, J = 2.8 Hz, 1H), 4.80 (s, 1H), 4.74-4.69 (m, 1H), 4.67-4.62 (m, 1H), 4.59-4.51 (m, 2H), 4.36 – 4.25 (m, 2H), 4.06 (d, J = 7.2 Hz, 1H), 3.78-3.68 (m , 2H), 2.89 (p, J = 7.0 Hz, 1H), 2.56 (s, 2H), 2.41 – 2.22 (m, 4H), 2.07 – 1.74 (m, 6H), 1.65 (s, 2H), 1.27 (dd, J = 6.8, 0.8 Hz, 6H), 0.65 (s, 2H), 0.43 (s, 2H). LC-MS: m/z 794.90 [M+H] + . Compound 45 1-((1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl )-8-fluoro-2-((1- (morpholinomethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin -4-yl)-3,8-diaza- bicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroethan-1-one. 1 H NMR (400 MHz, DMSO-d6) δ 10.18 (s, 1H), 9.08 – 9.01 (m, 1H), 7.98 (dd, J = 9.2, 5.9 Hz, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.40 (d, J = 2.6 Hz, 1H), 7.18 (t, J = 2.5 Hz, 1H), 4.80 (s, 1H), 4.72 (d, J = 13.0 Hz, 1H), 4.65 (s, 1H), 4.52 (d, J = 12.8 Hz, 1H), 4.36 – 4.25 (m, 2H), 3.94 (d, J = 6.4 Hz, 1H), 3.76 (d, J = 12.8 Hz, 1H), 3.68 (d, J = 13.1 Hz, 1H), 3.53 (t, J = 4.6 Hz, 4H), 2.39 (s, 4H), 2.31 (s, 2H), 2.00 – 1.86 (m, 4H), 0.65 (q, J = 3.1 Hz, 2H), 0.43 (q, J = 4.0 Hz, 2H). LC-MS: m/z 708.80 [M+H] + . Compound 46 ((5-ethynyl-6-fluoro-4-(8-fluoro-4-((1R,5S)-8-(2,2,2-trifluo roacetyl)-3,8- diazabicyclo[3.2.1]octan-3-yl)-2-((1-((4-(trifluoromethyl)pi peridin-1-yl)methyl) cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-7-yl)naphthalen-2 -yl)oxy)methyl dihydrogen phosphate. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.07 (s, 1H), 8.07 (dd, J = 9.2, 5.9 Hz, 1H), 7.80 (d, J = 2.7 Hz, 1H), 7.54 (t, J = 9.0 Hz, 1H), 7.36 (t, J = 2.3 Hz, 1H), 5.64 (d, J = 11.1 Hz, 2H), 4.79 (s, 1H), 4.72 (d, J = 12.9 Hz, 1H), 4.62 (s, 1H), 4.54 (d, J = 12.9 Hz, 1H), 4.32 (q, J = 11.2 Hz, 2H), 3.96 (d, J = 6.3 Hz, 1H), 3.75(dd, J = 22.9, 12.8 Hz, 2H), 3.72 (dd, J = 22.9, 12.8 Hz, 2H), 3.19 (dd, J = 22.9, 12.8 Hz, 1H), 3.05 (dd, J = 22.9, 12.8 Hz, 2H), 2.6 (dd, J = 22.9, 12.8 Hz, 2H) 1.88 (s, 6H), 1.55 (s, 2H), 0.74 (s, 1H), 0.70 (s, 2H), 0.55 (s, 2H). LC-MS: m/z 884.75 [M+H] + . Compound 47 5-ethynyl-6-fluoro-4-(8-fluoro-2-((1-((4-fluoropiperidin-1-y l)methyl)cyclopropyl) methoxy)-4-(8-(2,2,2-trifluoroacetyl)-3,8-diazabicyclo[3.2.1 ]octan-3-yl)pyrido[4,3-d] pyrimidin-7-yl)naphthalen-2-yl (tert-butoxycarbonyl)glycinate. 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.07 (d, J = 1.7 Hz, 1H), 8.25 (dd, J = 9.2, 5.9 Hz, 1H), 8.01 (d, J = 2.5 Hz, 1H), 7.67 (t, J = 9.0 Hz, 1H), 7.48 (d, J = 4.5 Hz, 2H), 4.80 (s, 1H), 4.76-4.68 (m, 1H), 4.65 (s, 1H), 4.58-4.51 (m, 1H), 4.46 (d, J = 4.8 Hz, 1H), 4.38 (t, J = 5.6 Hz, 1H), 4.35 – 4.24 (m, 2H), 4.09 – 3.98 (m, 2H), 3.80 – 3.66 (m, 2H), 3.30 (s, 3H), 2.52 (d, J = 2.6 Hz, 2H), 2.39 – 2.21 (m, 1H), 2.07 – 1.71 (m, 6H), 1.65 (s, 2H), 1.40 (s, 9H), 0.66 (s, 2H), 0.43 (s, 2H). LC-MS: m/z 881.90 [M+H] + . Compound 48 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-((1-((6-fluoro-3- azabicyclo[3.1.0]hexan-3-yl)methyl)cyclopropyl)methoxy)-5-(m ethoxy-d3)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-t rifluoroethan-1-one. 1 H NMR (400 MHz, DMSO-d6) δ 10.16 (s, 1H), 7.98 (dd, J = 9.2, 5.9 Hz, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.42 – 7.32 (m, 1H), 7.22 (s, 1H), 4.75 (s, 1H), 4.70 – 4.47 (m, 2H), 4.24 (q, J = 10.6 Hz, 4H), 3.96 (s, 1H), 3.56 – 3.38 (m, 2H), 3.05 (t, J = 9.4 Hz, 2H), 2.40 – 2.21 (m, 5H), 1.97 (s, 1H), 1.92 – 1.83 (m, 2H), 1.82 – 1.68 (m, 2H), 0.58 (s, 2H), 0.39 (s, 2H). LC- MS: m/z 756.10 [M+H] + . Compound 49 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-((2-((6-fluoro-3- azabicyclo[3.1.0]hexan-3-yl)methyl)allyl)oxy)-5-(methoxy-d3) pyrido[4,3-d]pyrimidin- 4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-trifluoroetha n-1-one. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.17 (s, 1H), 7.98 (dd, J = 9.2, 5.9 Hz, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.39 (d, J = 2.6 Hz, 1H), 7.26 – 7.20 (m, 1H), 5.11 (d, J = 10.9 Hz, 2H), 4.87 (s, 2H), 4.80 – 4.50 (m, 3H), 4.45 – 4.07 (m, 2H), 3.96 (d, J = 2.5 Hz, 1H), 3.57 – 3.43 (m, 2H), 3.05 (s, 2H), 3.00 (dd, J = 9.0, 2.9 Hz, 2H), 2.30 (d, J = 8.8 Hz, 2H), 1.95 (s, 1H), 1.86 (d, J = 12.7 Hz, 2H), 1.82 – 1.75 (m, 3H). LC-MS: m/z 742.1 [M+H] + . Compound 50 1-((1R,5S)-3-(2-((1-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)c yclopropyl)methoxy)-7-(8- ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-5-methox ypyrido[4,3-d] pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]oct-6-en-8-yl)-2,2,2- trifluoroethan-1-one. 1 H NMR (DMSO-d 6 , 400 MHz) δ 10.15 (s, 1H), 7.97 (dd, J = 9.2, 5.9 Hz, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.39 (d, J = 2.6 Hz, 1H), 7.21 (d, J = 2.5 Hz, 1H), 6.37 – 6.15 (m, 2H), 5.04 (d, J = 18.0 Hz, 2H), 4.42 – 4.32 (m, 1H), 4.26 – 4.12 (m, 2H), 4.05 – 4.02 (m, 1H), 3.94 (dd, J = 5.9, 1.0 Hz, 1H), 3.89 (d, J = 1.3 Hz, 3H), 3.67-3.54 (m, 2H), 2.96 (t, J = 8.0 Hz, 2H), 2.44 – 2.29 (m, 2H), 2.24 (d, J = 8.6 Hz, 2H), 1.32 (dt, J = 6.7, 2.6 Hz, 2H), 0.57 (q, J = 4.4, 3.3 Hz, 3H), 0.39 (q, J = 4.2 Hz, 2H), 0.26 (td, J = 7.7, 3.7 Hz, 1H). LC-MS: m/z 733.00 [M+H] + . Compound 51 1-(3-(2-((1-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)cycloprop yl)methoxy)-7-(8- ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-5-methox ypyrido[4,3-d] pyrimidin-4-yl)-6,6-difluoro-3,8-diazabicyclo[3.2.1]octan-8- yl)-2,2,2-trifluoroethan-1- one. 1 H NMR (400 MHz, DMSO-d6) : δ 10.19 (d, J = 13.0 Hz, 1H), 7.99 (dd, J = 9.4, 5.5 Hz, 1H), 7.48 (t, J = 9.0 Hz, 1H), 7.40 (t, J = 3.3 Hz, 1H), 7.28 (s, 1H), 5.08 – 4.88 (m, 2H), 4.86 – 4.63 (m, 1H), 4.34 – 4.18 (m, 2H), 4.17 – 4.01 (m, 1H), 3.95 – 3.89 (m, 3H), 3.82 – 3.61 (m, 2H), 2.94 (d, J = 7.8 Hz, 2H), 2.71 – 2.63 (m, 2H), 2.41 (d, J = 11.5 Hz, 1H), 2.35 (s, 1H), 2.23 (s, 2H), 1.31 (s, 2H), 1.23 (s, 1H), 0.57 (s, 3H), 0.39 (s, 2H), 0.24 (s, 1H). LC- MS: m/z 771.30 [M+H] + . Compound 52 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-2-(((2S,7aR)-2- fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5-(methoxy -d3)pyrido[4,3-d] pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-tri fluoroethan-1-one. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.16 (s, 1H), 7.97 (dd, J = 6.0, 8.9 Hz, 1H), 7.47 (t, J = 9.0 Hz, 1H), 7.39 (d, J = 2.6 Hz, 1H), 7.22 (s, 1H), 5.37-5.19 (m, 1H), 4.82-4.71 (m, 1H), 4.68-4.57 (m, 1H), 4.29-4.19 (m, 1H), 4.13 (d, J = 9.8 Hz, 1H), 4.06-4.00 (m, 1H), 3.97 (d, J = 2.8 Hz, 1H), 3.52-3.37 (m, 3H), 3.12-3.06 (m, 2H), 3.01 (s, 1H), 2.86-2.78 (m, 1H), 2.13 (d, J = 3.0 Hz, 1H), 2.05 (s, 1H), 2.01-1.95 (m, 2H), 1.89-1.75 (m, 6H). LC-MS: m/z 730.6 [M+H] + . Compound 53 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-5-(methoxy-d3)-2- ((1-((4-(trifluoromethyl)piperidin-1-yl)methyl)cyclopropyl)m ethoxy)pyrido[4,3-d] pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-tri fluoroethan-1-one. 1 H NMR (DMSO-d6, 400 MHz) δ 10.25-10.13 (m, 1H), 8.01-7.95 (m, 1H), 7.51-7.44 (m, 1H), 7.41- 7.37 (m, 1H), 7.24-7.19 (m, 1H), 4.79-4.71 (m, 1H), 4.67-4.56 (m, 1H), 4.30 (s, 2H), 3.96 (s, 1H), 3.47-3.40 (m, 2H), 3.07-2.97 (m, 3H), 2.34 (m, 2H), 2.05-1.93 (m, 2H), 1.92-1.82 (m, 4H), 1.77-1.70 (m, 3H), 1.43-1.20 (m, 3H), 0.64 (s, 2H), 0.48 (s, 2H). LC-MS: m/z 808.5 [M+H] + . Compound 54 1-(3-(5-cyclopropoxy-7-(8-ethynyl-7-fluoro-3-hydroxynaphthal en-1-yl)-8-fluoro-2- ((1-((4-(trifluoromethyl)piperidin-1-yl)methyl)cyclopropyl)m ethoxy)pyrido[4,3-d] pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-tri fluoroethan-1-one. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.20 (s, 1H), 7.98 (dd, J = 6.2, 9.2 Hz, 1H), 7.52-7.36 (m, 2H), 7.26-7.17 (m, 1H), 4.76-4.68 (m, 1H), 4.59-4.49 (m, 1H), 4.29 (s, 3H), 3.94 (d, J = 8.4 Hz, 1H), 3.85-3.75 (m, 2H), 3.54-3.45 (m, 4H), 3.28-3.21 (m, 2H), 3.03-2.94 (m, 2H), 2.18-2.05 (m, 2H), 1.97-1.66 (m, 8H), 0.91-0.65 (m, 8H). LC-MS: m/z 831.7 [M+H] + . Compound 55 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-5-isopropoxy-2- ((1-((4-(trifluoromethyl)piperidin-1-yl)methyl)cyclopropyl)m ethoxy)pyrido[4,3-d] pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2,2,2-tri fluoroethan-1-one. 1 H NMR (DMSO-d6, 400 MHz) δ 10.22 (s, 1H), 7.97 (dd, J = 6.0, 9.2 Hz, 1H), 7.50-7.43 (m, 1H), 7.38 (d, J = 2.4 Hz, 1H), 7.19 (dd, J = 2.4, 8.8 Hz, 1H), 5.39-5.30 (m, 1H), 4.75 (s, 1H), 4.65-4.53 (m, 1H), 4.34-4.28 (m, 2H), 3.95 (d, J = 3.4 Hz, 1H), 3.50-3.26 (m, 4H), 3.03- 2.97 (m, 2H), 2.38-2.31 (m, 1H), 2.30-2.24 (m, 1H), 2.23-2.13 (m, 1H), 1.97-1.65 (m, 8H), 1.40-1.34 (m, 1H), 1.33-1.26 (m, 6H), 0.64 (s, 2H), 0.41 (s, 2H). LC-MS: m/z 833.7[M+H] + . Compound 56 1-(3-(7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluo ro-5-(2,2,2-trifluoro- ethoxy)-2-((1-((4-(trifluoromethyl)piperidin-1-yl)methyl)cyc lopropyl)methoxy)pyrido [4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-2, 2,2-trifluoroethan-1-one. 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.29-10.12 (m, 1H), 8.04-7.96 (m, 1H), 7.48 (t, J = 9.0 Hz, 1H), 7.41 (d, J = 2.6 Hz, 1H), 7.23 (dd, J = 2.4, 7.1 Hz, 1H), 5.17-4.91 (m, 2H), 4.81-4.70 (m, 1H), 4.63-4.52 (m, 1H), 4.33 (s, 2H), 4.00 (d, J = 7.8 Hz, 1H), 3.60-3.41 (m, 2H), 3.05- 2.93 (m, 2H), 2.39-2.15 (m, 4H), 1.93-1.68 (m, 8H), 1.42-1.21 (m, 3H), 0.65 (s, 2H), 0.43 (s, 2H). LC-MS: m/z 873.7 [M +H] + . Additional compounds which are contemplated and part of the present disclosure include the following.
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. Biological Assays/Testing Cell lines The following cancer cell lines were employed: A427 human lung carcinoma [1 copy of KRAS(G12D)] (ATCC, #HTB-53); AGS human gastric adenocarcinoma [1 copy of KRAS(G12D)] (ATCC, #CRL-1739); and AsPC-1 human adenocarcinoma [2 copies of KRAS(G12D)] (ATCC, CRL-1682). Cell lines were cultured essentially according to American Type Culture Collection (ATCC) recommendations. Cell Cytotoxicity A427 and AGS cells were plated in 96-well tissue culture plates at 4,000 cells/well and incubated at 37°C/5% CO2 for 72 hr in 100 μl of media.3-fold serial dilutions of each test compound were prepared ranging from 20 μM to 1.02 nM. Each cell line was then treated with test compounds at various concentrations with a final concentration of 0.5% DMSO/well, and then incubated at 37°C/5% CO2 for 24 hr.100 μl of CellTiter-Glo® Reagent (Promega Corporation, Madison, WI) was added to each well and processed according manufacturer’s protocol. Results were analyzed and IC50 values were calculated in GraphPad 7 software. Results are listed in Table 1. KRAS(G12D)/SOS1 homogeneous time-resolved fluorescence (HTRF) assay Binding of test compounds to KRAS(G12D) target protein, which in turn blocks KRAS(G12D) interaction with the SOS1 protein, was measured in the absences of GTP by homogeneous time-resolved fluorescence using the KRAS-G12D/SOS1 Binding Assay Kit (Cisbio, #63ADK000CB16PEG), following the manufacturer’s instructions, except as noted.3-fold serial dilutions of each test compound were prepared ranging from 20 μM to 1.02 nM. The test compound was mixed and incubated with reaction components, incubated in a sealed plate at 4°C for 3 hr and fluorescence was measured using a PerkinElmer Envision plate reader. The KRAS(G12D)-SOS1 IC50 values (the concentration of 50% of the maximal inhibition) were calculated using GraphPad Prism 7 software. Results are listed in Table 1. Table 1 - Cytotoxicity and Target Binding Assays
1 CellTiter-Glo cytotoxicity assay for A427 and AGS cells: A: 1-100 nM, B: 101-1000 nM, C: 1001-10000 nM, D: >10,000 nM; 2 KRAS(G12D)-SOS1 HTRF binding. A: 1-100 nM, B: 101-1000 nM, C: 1001-10000nM, D: >10,000 nM. Oral Bioavailability CD-1 mice were randomly assigned to 6 groups with 3 male mice in each compound group. Control groups included Compound 1 (50 mg/kg), Compound 3 (25 mg/kg), and Compound 5 (25 mg/kg). Trifluoroacetate (COCF3) groups included Compound 2 (50 mg/kg), Compound 4 (50 mg/kg) and Compound 6 (50 mg/kg). Compounds were orally administered (PO) in a single dose to each mouse in its group. Blood samples were taken within 72 hours. Bioavailability (F%) was determined and by liquid chromatography-mass spectrometry (LC-MS/MS). The mean oral %F is provided in Table 2. Table 2
In Vivo Xenograft Tumor Model BALB/c nude mice at 8 to 9 weeks age were maintained in a pathogen-free environment. AsPC-1 cells (5×10 6 ) were subcutaneously implanted into female mice. Mice bearing established tumors (~ 170 mm3) were randomized into treatment groups of 4. Mice were intravenously (i.v.) dosed with 5% sulfobutylether-beta-cyclodextrin (SBECD) vehicle control once a week (QW) or orally dosed (PO) twice a day (BID) with 75 mg/kg Compound 3, for over 3 weeks. Tumors were measured twice a week with calipers. Tumor volume (TV) was calculated as TV = (L×W2)/2, where L is tumor length and W is tumor width. Results were graphed with +/- standard error (SEM). Student T-test with Bonferroni post hoc adjustment was used to measure statistical significance of the final (day 24) tumor measurements (P=0.036). Results are shown in FIG 1. Although the disclosure has been described in connection with specific embodiments, it should be understood that the disclosure as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the disclosure are intended and understood by those skilled in the relevant field in which this disclosure resides to be within the scope of the disclosure as represented by the following claims. All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent and publication was specifically and individually indicated to be incorporated by reference.
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