KRAS g12d degradation agent as well as preparation method therefor and use thereof
A KRAS G12D degrader compound targets and degrades KRAS G12D protein, overcoming drug resistance and administration challenges, offering a therapeutic solution for KRAS G12D-related cancers.
Patent Information
- Application Number
- AU2025258191
- Authority / Receiving Office
- AU · AU
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2025-04-11
- Filing Date
- 2025-04-18
- Publication Date
- 2026-11-05
AI Technical Summary
Current KRAS G12D inhibitors face challenges with drug resistance and require injection for administration, while existing KRAS G12D degraders are limited in efficacy.
Development of a KRAS G12D degrader compound, represented by formula I, which can degrade and/or inhibit the KRAS G12D protein in cells, utilizing specific structural components to target the KRAS G12D mutant effectively.
The compound effectively degrades and inhibits KRAS G12D protein, addressing drug resistance issues and providing a potential therapeutic option for KRAS G12D-related cancers.
Abstract
Description
[0001] The present application claims the right of the priority of the Chinese Patent Application No. 2024104786939 filed on April 19, 2024, Chinese Patent Application No. 2024114044653 filed on October 10, 2024, Chinese Patent Application No. 2025100771962 filed on January 17, 2025, and Chinese Patent Application No. 2025104624657 filed on April 11, 2025. The entire content of the above Chinese patent application is incorporated herein by reference. TECHNICAL FIELD
[0002] The present disclosure relates to the technical field of pharmaceuticals, and particularly relates to a KRAS G12D degrader, a preparation method therefor, and use thereof. BACKGROUND
[0003] RAS (rat sarcoma) is one of the oncogenes with the highest mutation rate in tumors, and its mutations are present in about 30% of human malignancies. The RAS family includes KRAS, NRAS, and HRAS, among which KRAS (Kirsten rat sarcoma viral oncogene) is more prone to mutations compared to the other two RAS subtypes. KRAS mutations account for approximately 85% of all RAS mutations and are particularly common in solid tumors (Cancer Res. 2020, 80, 2969-2974). The mutation of the KRAS gene exists in 30%-40% of colorectal cancers, 90% of pancreatic cancers, and 15%-20% of lung cancers. After KRAS is activated, it regulates functions such as cell proliferation, differentiation, and survival through downstream signaling pathways such as RAF-MEK-ERK and PI3K-AKT-mTOR. After the mutation of the KRAS gene, the protein remains continuously activated, resulting in sustained activation of downstream signaling pathways, and thus promoting tumorigenesis (Nat. Rev. Cancer 2015, 15, 290-301). Therefore, KRAS is an important therapeutic target in cancer treatment, with efforts focused on targeting the KRAS protein itself, its post-translational modifications, membrane localization, protein-protein interactions, and the downstream signaling pathways of RAS.
[0004] Since KRAS binds to GTP or GDP with ultra-high affinity (in the picomolar concentration level), and the surface of the RAS protein is smooth, lacking ideal binding sites for small molecules, the development of competitive inhibitors that act directly on the RAS protein is considered extremely challenging (Nat. Rev. Drug Discov. 2020, 19, 533-552).
[0005] At present, according to the action modes of inhibitors, KRAS inhibitors (Cancer Discov. 2022, 12, 924-937) can be divided into:
[0006] inhibitors that directly target KRAS, such as KRAS G12C, KRAS G12D, KRAS G12R (J. Am. Chem. Soc. 2022, 144, 35, 15916-15921), and KRAS G12S (Nat. Chem. Biol. 2022, 18, 1177-1183) inhibitors; and
[0007] inhibitors that act indirectly on KRAS, such as SHP2 inhibitors (SHP 099, Nature 2016, 535, 148152 & J. Med. Chem. 2016, 59, 7773-7782; RMC4550, Nat. Cell Biol. 2018, 20, 1064-1073; TNO155, J. Med. Chem. 2020, 63, 22, 13578-13594; RMC4630, WO 2021142026A1; JAB-3068, WO2017211303A1, etc.), SOS1 inhibitors (Bay-293, Proc. Natl. Acad. Sci. U S A. 2019, 116, 2551-2560; BI-3406, Cancer Discov. 2021, 11, 142-157; MRTX0902, J. Med. Chem. 2022, 65, 678-9690, etc.), and KRAS (on) inhibitors (WO2021091982A1, WO2022060836A1).
[0008] In recent years, with the continuous advancement in KRAS research, significant progress has been made in the development of KRAS inhibitors (Cancer Discov. 2022, 12, 924-937). In the design of KRAS-G12C inhibitors, the cysteine residue at codon 12 is primarily utilized due to its high propensity to form covalent bonds. The strategy of covalently targeting the cysteine site has become a crucial breakthrough in the development of KRAS drugs. Shokat discovered a new allosteric binding pocket in Switch-II, called the Switch-II pocket, and developed the first batch of KRAS-G12C covalent inhibitors (Nature 2013, 503, 548551; Nat. Rev. Drug Discov. 2016, 15, 771-785). On May 28, 2021, the FDA granted accelerated approval to Lumakras (Sotorasib, AMG510) (Nature 2019, 575, 217-223; J. Med. Chem. 2020, 63, 52-65), developed by Amgen, for the treatment of patients with non-small cell lung cancer harboring the KRAS-G12C mutation (N. Engl. J. Med. 2021, 384, 2371-2381). Currently, multiple small molecule drugs targeting KRAS-G12C are in clinical trials globally. On February 16, 2022, the FDA accepted the New Drug Application submitted by Mirati Therapeutics for its second KRAS-G12C inhibitor, Adagrasib (MRTX849) (Cancer Discov. 2020, 10, 54-71 & J. Med. Chem. 2020, 63, 6679-6693). This inhibitor is intended for the treatment of patients with nonsmall cell lung cancer harboring the KRAS G12C mutation who have received at least one systemic therapy (N. Engl. J. Med. 2022, 387, 120-131). Adagrasib is expected to become the second KRAS-targeted drug worldwide.
[0009] Unlike the KRAS G12C mutation, the most common mutation in patients with non-small cell lung cancer (NSCLC), KRAS G12D is the most common mutation in colorectal cancer and pancreatic cancer. The KRAS G12C mutant contains a cysteine (Cys) residue at codon 12, which facilitates the formation of covalent bonds. This structural characteristic enables the covalent binding of the mutant to small molecule inhibitors. In contrast, the KRAS G12D mutant has aspartic acid (Asp) at codon 12, which precludes the use of this covalent binding strategy.
[0010] The activation of KRAS-G12D is primarily mediated by protein-protein interactions. Considering the lack of distinct binding pockets on the surface of the KRAS protein and the high binding affinity of cyclic peptides for protein surfaces, peptide molecules are used as the first choice for targeting KRAS G12D (Biochem. Biophys. Res. Commun. 2017, 484, 605-611; Bioorg. Med. Chem. Lett. 2017, 27, 2757-2761; ACS Med. Chem. Lett. 2017, 8, 732-736; Sci. Rep. 2020, 10, 21671). At present, several potent cyclic peptides targeting KRAS G12D have been reported (MedChemComm 2013, 4, 378-382; Angew. Chem., Int. Ed. 2015, 54, 7602-7606; J. Med. Chem. 2021, 64, 13038), but they have relatively poor cell membrane permeability. The ability of cells to uptake peptides depends on the regulation by the cell surface receptor neuropilin-1 (NRP1), and this protein is overexpressed on the cell membrane of human lung cancer cells. Therefore, cyclic peptides targeting both NRP1 and KRAS G12D are picked out, and in this way, the cyclic peptides not only have relatively high cellular uptake ability, but also can act on KRAS G12D to exert anti-tumor effects (J. Am. Chem. Soc. 2022, 144, 7117-7128).
[0011] On December 10, 2021, Mirati Therapeutics reported the first non-covalent, potent, and highly selective KRAS-G12D inhibitor, MRTX1133 (J. Med. Chem. 2022, 65, 3923-3942; WO2021041671A1). MRTX1133 can inhibit KRAS-G12D mutant cells in an activated or inactivated state, but does not inhibit wild-type tumor cells, with a specificity of 1000-fold or higher. It demonstrated a dose-dependent inhibitory effect in in vivo xenograft tumor models of pancreatic cancer and colorectal cancer.
[0012] Several classes of KRAS G12D inhibitors are disclosed in patents such as WO2021041671A1, WO2022015375A1, WO2022031678A1, WO2022066646A1, WO2022098625A1, WO2022192790A1, WO2022192794A1, WO2021106231A1, WO2021107160A1, and WO2022173870A1.
[0013] Targeted protein degraders have become a globally popular research and development technology in recent years (Nat. Rev. Drug Discov. 2017, 16, 101-114; Nat. Rev. Drug Discov. 2019, 18, 949-963; Nat. Rev. Drug Discov. 2022, 21, 181-200). This technology has the following characteristics: the development of “druggable” targets has encountered bottlenecks, while “undruggable” targets hold immense potential; the PROTAC technology is expected to address the development challenges of “undruggable” targets; PROTAC drugs offer advantages such as good druggability, high selectivity, ability to overcome drug resistance, diverse routes of administration, low dose, and low toxicity.
[0014] Therefore, there is an unmet clinical need for KRAS G12D targeted protein degraders. SUMMARY
[0015] The technical problem to be solved in the present disclosure is to overcome the drawbacks that KRAS G12D inhibitors are prone to drug resistance and KRAS G12D degraders that have entered clinical trials need to be injected for administration. For this purpose, the present disclosure provides a KRAS G12D degrader, a preparation method therefor, and use thereof. The compound provided by the present disclosure can effectively degrade and / or inhibit the KRAS G12D protein in cells.
[0016] The present disclosure provides a compound of formula I, and / or a stereoisomer, an enantiomer, a diastereomer, an atropisomer, a deuteride, a hydrate, a solvate, or a prodrug thereof, and / or a pharmaceutically acceptable salt thereof: G-L-E I
[0017] wherein G is G1 or G2;
[0018] in G1,
[0019] X is C or N;
[0020] R1 is H or halogen;
[0021] R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl;
[0022] Ra is H, fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl is optionally substituted with 1 or more halogen, OH, or deuterium;
[0023] Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1- 3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;
[0024] in G2,
[0025] W is N or CRza;
[0026] Rza is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy;
[0027] Z is N or CRzb;
[0028] Rzb is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy; Rzc is phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole,
[0029] , or pyridine, wherein the phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, H N^O . Cj .... . . . indazole, , , or pyridine is optionally substituted with 1 or more R1;
[0030] Rzd is hydrogen, deuterium, methyl, deuterated methyl, halomethyl, -CD2F, -CDF2, C2-6 alkyl, or C3-6 cycloalkyl, wherein the methyl, C2-6 alkyl, or C3-6 cycloalkyl is optionally substituted with 1 or more hydroxy, deuterium, or halogen;
[0031] each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, -C(O)NR23R24, CH2R25, N=S(O)(C1-6 alkyl)2, S(O)C1-6 alkyl, S(O)2R26, -S-C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-6 hydroxyalkynyl, C1-6 cyanoalkyl, triazolyl, -S-C1-6 haloalkyl, C1-6 hydroxyalkyl, CH2C(O)NR27R28, -C2-6 alkynyl-NR29R30, C2-6 deuterated alkynyl, (C1-6 alkoxy)C1-6 haloalkyl-, or cycloalkyl, wherein the cycloalkyl is optionally substituted with 1 or more halogen or C1-6 alkyl;
[0032] or R1' is SF5;
[0033] R21 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C(O)C1-6 alkyl;
[0034] R22 is hydrogen, C1-6 alkyl, or C1-6 haloalkyl;
[0035] or R21 and R22, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0036] R23 and R24 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0037] or R23 and R24, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0038] R25 is hydroxy, cyano, heterocyclyl, NR25aR25b, C(O)NR25cR25d, or SO2C1-6 alkyl;
[0039] R25a, R25b, R25c, and R25d are each independently H or C1-6 alkyl;
[0040] or R25a and R25b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0041] or R23c and R25d, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0042] R26 is C1-6 alkyl, C1-6 haloalkyl, or NR26aR26b;
[0043] R26a and R26b are each independently H or C1-6 alkyl;
[0044] or R26a and R26b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0045] R27 and R28 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0046] or R27 and R28, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0047] R29 and R30 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0048] or R29 and R30, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0049] R3a is hydrogen, halogen, hydroxy, cyano, C1-3 alkyl, C1-3 deuterated alkyl, or C1-3 haloalkyl;
[0050] n is 0, 1, 2, 3, 4, or 5; or
[0053] The present disclosure provides a compound of formula I, and / or a stereoisomer, an enantiomer, a diastereomer, an atropisomer, a deuteride, a hydrate, a solvate, or a prodrug thereof, and / or a pharmaceutically acceptable salt thereof: G-L-E
[0054] wherein G is G1 or G2;
[0055] in G1,
[0056] X is C or N;
[0057] R1 is H or halogen;
[0058] R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl;
[0059] Ra is H, fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl is optionally substituted with 1 or more halogen, OH, or deuterium;
[0060] Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1-3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;
[0061] in G2,
[0062] W is N or CRza;
[0063] Rza is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy;
[0064] Z is N or CRzb;
[0065] Rzb is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy; Rzc is phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole,
[0066] , or pyridine, wherein the phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, H . J . . . . . . , indazole, , , or pyridine is optionally substituted with 1 or more R1';
[0067] Rzd is hydrogen, deuterium, methyl, deuterated methyl, halomethyl, -CD2F, -CDF2, C2-6 alkyl, or C3-6 cycloalkyl, wherein the methyl, C2-6 alkyl, or C3-6 cycloalkyl is optionally substituted with 1 or more hydroxy, deuterium, or halogen;
[0068] each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, -C(O)NR23R24, CH2R25, N=S(O)(C1-6 alkyl)2, S(O)C1-6 alkyl, S(O)2R26, -S-C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-6 hydroxyalkynyl, C1-6 cyanoalkyl, triazolyl, -S-C1-6 haloalkyl, C1-6 hydroxyalkyl, CH2C(O)NR27R28, -C2-6 alkynyl-NR29R30, C2-6 deuterated alkynyl, (C1-6 alkoxy)C1-6 haloalkyl-, or cycloalkyl, wherein the cycloalkyl is optionally substituted with 1 or more halogen or C1-6 alkyl;
[0069] R21 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C(O)C1-6 alkyl;
[0070] R22 is hydrogen, C1-6 alkyl, or C1-6 haloalkyl;
[0071] or R21 and R22, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0072] R23 and R24 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0073] or R23 and R24, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0074] R25 is hydroxy, cyano, heterocyclyl, NR25aR25b, C(O)NR25cR25d, or SO2C1-6 alkyl;
[0075] R25a, R25b, R25c, and R25d are each independently H or C1-6 alkyl;
[0076] or R25a and R25b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0077] or R23c and R25d, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0078] R26 is C1-6 alkyl, C1-6 haloalkyl, or NR26aR26b;
[0079] R26a and R26b are each independently H or C1-6 alkyl;
[0080] or R26a and R26b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0081] R27 and R28 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0082] or R27 and R28, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0083] R29 and R30 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0084] or R29 and R30, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0085] R3a is hydrogen, halogen, hydroxy, cyano, C1-3 alkyl, C1-3 deuterated alkyl, or C1-3 haloalkyl;
[0086] n is 0, 1, 2, 3, 4, or 5; o aZDXD Pn^e ZA^Ak^^k k^^xk N^^kx^x 0 A A __A^j-k ^° / \ N 1 k^Ak k / N^kx^ D D O A / V ^P / Ve ■^O / x N ] k / N^k\^x D D 0 A Z V V k^N A Ao'^kk^N^k k^^k k^N^xk^X D D O a z V k^N^ Ao-^y^N^ kik k^xN^^k^O a / y__pk ' o Y N > kkxx kxNxkxx"0 0 A / / \ / \ zx x'xk^k Z 0 AP N > [ '"xk k^xNx kx^O D D O az ^rvSk ^° I] N | | ^\k k^N^Jx^O O A / <^N^Y b / ^°^kk^N^k k^X / k kxNxk / R XX XX XX O * a z Ay pyke z 0 / \ N | k 'x^k \xN\kx , , 9 B AZXDlkD Pn^Zb z^qN^k k^^-k k^N^kx^x- , , 0 3 azdxd r^B nr^'^k k^N^ / kx D D , , / 0 B a / v__xxrVkB Z 0 N | ^X^k k^N^kx / 0 , , o AZ x'-'x / 'x X-X xxk^'k^k B V z O Y N A A ^x J Z B k^ / N.Jx^O D D , , 0 a / „ yx / AVa z 0 / \ N k ll\k k^ / N^kx^O , , 0 a / ^N^yB ^0]]^ k^X^k k / N^kxx , , 0 3 AZ / x / X XX XX B z ° k N i k 'xk 11 ^N^ok , , 0 a / ^N^y / g k^'^y kc k^N^kx / O , h , or diastereomer, an atropisomer, a deuteride, a hydrate, a solvate, or a prodrug thereof, and / or a pharmaceutically acceptable salt thereof: G-L-E
[0090] wherein G is G1 or G2;
[0091] in G1,
[0092] X is C or N;
[0093] R1 is H or halogen;
[0094] R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl;
[0095] Ra is H, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl is optionally substituted with 1 or more halogen, OH, or deuterium;
[0096] Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1-3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;
[0097] in G2,
[0098] W is N or CRza;
[0099] Rza is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy;
[0100] Z is N or CRzb;
[0101] Rzb is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy;
[0102] Rzc is phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole, or pyridine, wherein the phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole, or pyridine is optionally substituted with 1 or more R1';
[0103] Rzd is hydrogen, deuterium, methyl, deuterated methyl, halomethyl, -CD2F, or -CDF2;
[0104] each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, -C(O)NR23R24, CH2R25, N=S(O)(C1-6 alkyl)2, S(O)C1-6 alkyl, S(O)2R26, -S-C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-6 hydroxyalkynyl, C1-6 cyanoalkyl, triazolyl, -S-C1-6 haloalkyl, C1-6 hydroxyalkyl, CH2C(O)NR27R28, -C2-6 alkynyl-NR29R30, C2-6 deuterated alkynyl, (C1-6 alkoxy)C1-6 haloalkyl-, or cycloalkyl, wherein the cycloalkyl is optionally substituted with 1 or more halogen or C1-6 alkyl;
[0105] R21 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C(O)C1-6 alkyl;
[0106] R22 is hydrogen, C1-6 alkyl, or C1-6 haloalkyl;
[0107] or R21 and R22, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0108] R23 and R24 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0109] or R23 and R24, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0110] R25 is hydroxy, cyano, heterocyclyl, NR25aR25b, C(O)NR25cR25d, or SO2C1-6 alkyl;
[0111] R25a, R25b, R25c, and R25d are each independently H or C1-6 alkyl;
[0112] or R25a and R25b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0113] or R23c and R25d, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0114] R26 is C1-6 alkyl, C1-6 haloalkyl, or NR26aR26b;
[0115] R26a and R26b are each independently H or C1-6 alkyl;
[0116] or R26a and R26b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0117] R27 and R28 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0118] or R27 and R28, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0119] R29 and R30 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;
[0120] or R29 and R30, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;
[0121] R3a is hydrogen, halogen, hydroxy, cyano, C1-3 alkyl, C1-3 deuterated alkyl, or C1-3 haloalkyl;
[0122] n is 0, 1, 2, 3, 4, or 5; A / X^ ■■‘Xxx'' n ^x* xA
[0123] L is i 0 A / X^V B x^n-Xv / CN , 0 a / r^N^x b Xy>; n^X Xx J x°...... OH , 0 A / XX^Xb 'O X'nX fX J T X-AA^ CN , 0 A / ^"°xnX X^\X kX\X\X , 0 a? x^ z ° xx N ] । xX ^N^0J , 0 o ^Xb az XX'Xb F F , , 0 AX / \ xx^?^XB XxJ A Mn^X , 0 a? ^° xx n x X^n-^X / F , 0 AA / \ / x x\ C^B z o r n X < \ J ......1......9-°...... HO , 0 a? X"?^Xb ^° XX N i X^\X X / N^X\^o , 0 A / X^n^Vb x.....G......cP , B B , and L is linked to G via A or B; , or
[0124]
[0125] 0 . In some preferred embodiments of the present disclosure, the compound of formula I is not any one of the compounds disclosed in WO2024083258A1 (International Application No. PCT / CN2023125979), specifically not any one of the compounds disclosed on pages 108-126.
[0126] In some preferred embodiments of the present disclosure, X is N.
[0127] In some preferred embodiments of the present disclosure, R1 is halogen.
[0128] In some preferred embodiments of the present disclosure, R2 is C2-C4 alkynyl.
[0129] In some preferred embodiments of the present disclosure, Ra is fluorine, C1-3 alkyl, C1-3 alkyl-O-C1- 3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium; preferably, Ra is C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium.
[0130] In some preferred embodiments of the present disclosure, in Ra, the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more deuterium.
[0131] In some preferred embodiments of the present disclosure, Rb is H.
[0132] In some preferred embodiments of the present disclosure, Rc is halogen.
[0133] In some preferred embodiments of the present disclosure, L is or
[0134] In some preferred embodiments of the present disclosure, E is ; preferably, E is
[0135] In some preferred embodiments of the present disclosure, in the compound of formula I,
[0139] R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl;
[0140] Ra is C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium;
[0141] Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1- 3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;
[0142] L is B , and L is linked to G via A; 0 0 o .
[0143] E is 0 , or or
[0144] In some preferred embodiments of the present disclosure, in the compound of formula I,
[0148] R2 is C2-C4 alkynyl;
[0149] Ra is C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more deuterium;
[0150] Rb is H;
[0151] Rc is halogen;
[0152] L is is linked to G via A; or and L
[0153]
[0154] cd3 0 In some preferred embodiments of the present disclosure, in the compound of formula I,
[0158] R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl;
[0159] Ra is fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl- O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium;
[0160] Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1- 3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;
[0166] R1 is halogen;
[0167] R2 is C2-C4 alkynyl;
[0168] Ra is fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl- O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more deuterium;
[0169] Rb is H;
[0170] Rc is halogen; o A / Yn^YB A / 1 YN * Y , 0 AZ kA^Ye A / ^O Y N I Z o [ N Y < k / NxY"Y YN^k\ / , 0 A / Y^Ak A / ^0 Y N Y [ X\Y Z 0 N OMe k^N^kxk OMe k / , 0 a,v PnYb az V n^k r^v / 1 yyv kY-^k\k k , o A / Y^V B ZO Y N I Y'^-'Y A / DY kY\k\k ^'O^Y'N^ dD Y , 0 A Z , Yb a ^OTY z 0 Yf N l । Y Y YN\k\^ , 0 . D D Yuk / A / DY aA X / \ / \ । Xb az v zo Y^ n y [ x\ / z 0 i N kY\k\Y* * Y , o A A B z o y^ N । [ ^\Y D d k / N^Yk^o 'XkkkYhK DZ D Y , S A / __ az^5<° ^Yb AtOTY < Y / \ NY irY ^nyAx' k / N. A ,0 D 0 , or to G via A; <Yy' , 0 y3n^b , 0 YkT^JN^^B NxY\^ , > Y- , , Y'- .Nxk-X / , O kSO^B - N\Yx^ D D , -, Y-xNxYxY , 'i Yk7" , YNke / 0 , and L is linked E is
[0172]
[0173] preferred embodiments of the present disclosure, G CD3 OH OH OH OH In some OH OH H N. / CD N i hi r Nn v- H N .CD-3 H N / .CD3 OH H N ,CD3 ~N Nx OH t.....’ OH OH In some preferred embodiments of the present disclosure, G is
[0174] N H N. OH or
[0175] In some preferred embodiments of the present disclosure, G is , and L is
[0176] In some preferred embodiments of the present disclosure, G is OH or
[0177] In some preferred embodiments of the present disclosure, G is OH , and L is o a? A / Z ° N AA ■ Ax / 'y A F3A-N 1 1 , 0 A / -x A / A-A^A A / A ^B i a^'A'A^'A A^'^A i \ AnA-A c , 0 a / Ak^Ae ZO N < \J * AN-A^ A / 1 Ao\f N HC> A , 0 A / A^ / A A / AtA * ANA^° * A , O A / A^ B A / qAAn^] A'-xA z''^ A Nx / kzO k / N" , 0 a / -,-, / ^ b a^Y A An-Ax / .
[0178] In some preferred embodiments of the present disclosure, G is ° AZ — — —, d d --, A / ^cr y n a / VA A^A^ 1 f ■-o'^ i"~\A z, / n,,A-,x , 0 y rSC^^6 / N^k / ° , ., -¼ / N^Ax / N , o kSC^^6 -N\A\ / ° , 0 ^AT^jN^kz B / N^A'', / 0 , 0 rSO^6 , or A aJ / xA (A jfAft xjxiN^ OH , and L is -, rA N\Ax^ D D , o A / D\ / D ' o t n i X x J xC >< 1 'x^AA ^O^f N D D A / , 0 a / ^Ab aZ ' O Y N A A z ° / \ N ANxX? A D D , 0 A / V ^N^ / g AAAY AAy^N^A a^xA 1 ^x / NxXX^O , ? A / A / V 7B "aTi aaA'n'^'''] A / NxxAxO , o A / B A / .. ,. z O N y X X.Y ' ° A N , o A / ^N^y B A / N^ , o A / B yy\^ .
[0179] In some preferred embodiments of the present disclosure, G is ° Ay / X / X / X d d xx A / ^o^ yy n A / VD f N / B 1 f z'x'y A"" xx l^^N^ / XX 0 'i rS^^3 N\xAx , pAyA xNxAx^ D D , o "i aOnA / b / N^xA^O D D , o aSOnA)Zb -nAxx° D^ D , o ^''A^A b N^X\^ , o A''n^'A b '''XO , or A f j r y xX N^y'-N xyiN^ OH , and L is i ox N\XXX D D o A / .^ A / D\ / D ' o t n > r J xC >< xx 1 ^N.xj° A ? D D \ / , 0 a / aZ ' O V N > | z ° / \ N ^x / V? k, D D , 0 A / V ^N^\ / b y^xA 1 A kxNxAxx° , ? A / A / V A°QC? ^q / "7\ n^ A^N^Axx0 , o A / Ak^'k B A / xx xx z o n z ° / \ N , o A / ^N^y B A / ii\^ N^ , o A / B yy\^ . [
[0180] In some preferred embodiments of the present disclosure, G is O A / AZ^.%X CX / b A”Tr , 0 'i rS^^3 N-X\ / , yyy^ D D , o "i Hu^ D D , o rSO1^6 D^ D , o ^y ^y b , o y''N^y / b '''XO , or H ^n^cd3 yp, OH , and L is O D D , o V A / D D 1 O / C | D D \^r , AZ r^B aa x<A AxX x-^ k^N. Jx D D , 0 az °x° h / B aAyv Ao y^'N ^ kAxX / O , a / DvD PN 7B A°^C? k^N^Jx^O , o A / .Z'^Vb V'X'XXX Ao An ^ A^ 0 | l^^N^ / XX * \x , 0 A / A / ^° / \'NZ f^xx ^°7\ XN ] C L / N^ J k / N. Xx x / xz , or xx \ZX
[0181] In some preferred embodiments of the present disclosure, G is 0 A / pN^yB A / ^N^AxX , 0 ^xXXX , i rO^6 NxX\X D D , O xO Z; - Nx^x^O dA d , , ......[X -Nx^X / O D D , 0 NxxC / 0^ , 0 x^j^B ^° . H §A 0H , and L is 0 r^O^3 N^x\x , o 0 of the present disclosure, G is and
[0182] In some preferred embodiments or 0 D □ AZ x A / V ( N y B ^O'^y^N'^y * k k^N , 0 V ^C'l'ke Xv^ ,. ^Oyjy ^Cu \z , or i- (
[0183] In some preferred embodiments of the present disclosure, G is 0 A / ^y B A / • k / N^ J\ J is x^x^ , O Az k^N^y B a / ^o^y^ y^-^y k_ / N^X^O ‘ k , 0 A / y^N^YB A / ^o^yyy ry Ayyyy * * < , O A / C / X / X / X Az ..^ o'^y'^N"^ ypxk A''o'''yy''^N"'^ k, / N-vAzO k / N. , A / ^o'x^xy^'N'x^y ^Nx° B O ^^kNAy ^N^y^Y3 D^ D , o B . H I j ? 1 JuXa yyy^x OH , and L 0 y^^xy"^^6 x_ / N^y\^ , 0 z yy^6 .^N^yx^0 , 0 ^r^N'^yB xN^y .,,^0 , 0 ySO1^6 , or H 0
[0184] In some preferred embodiments of the present disclosure, G is B B
[0185] In some preferred embodiments of the present disclosure, G is and L and L is
[0186] In some preferred embodiments of the present disclosure, G is 0H , and L is O B or c
[0187] In some preferred embodiments of the present disclosure, G is o is ‘ '■ , k O AZ / . / xCb6 a / ° / \ y ] I k / N^A^o 1 L , 0 11 , A J A / \ n / b Z / x 0 N ^Ax^-O 1 \x , 0 A / B A / f[X / k^N.^ L^ , A / Uh a z v L- Ata^n"''^ B A / h , 0 U AZ Az yD r^N^yB ANXxA / , 9 A / DvD A Z AD 5<D AN / B ^oxy^n' / ^ Z^\A An-Ax / , H ; > OH , and L 0 A b^A^3 x-Nx-Ax-, 0 B , 0 ^T^n'^Vb 'N-A^ / O , 0 B '^^0 , 0^. JxA\ / , ., ......Xv. D D , 0 "^i rZCA8 D^ D , n n Jx A Z V Z0 ax xx >< xx b z ° N 1 [ i L k^N^xk^k D D , 0 A / V__^P^B A / “xX N^| I'v / kxNx-k\^k Y , o AZ x\x~- ^x B A D D D ^o V n > A Z V JkD k^N^xkY D D , o AZ^><° AZV Z ° / \ y | ( ^\k k^N^xkY Z\ D D , 0 AZ W r^N^R x\ / k\ x'^^x 1 r\ Z ° / \ N 1 [ A / DVD ^x / Nx^x / D D * k , o AZ XX XX XX xC / 'ke . z ° N I [ A / 'xN'^k'-x0 D D 11 k^ / N^ , o A / <^N^ / B n"i k l\xNx^\x F . {
[0188] In some preferred embodiments of the present disclosure, G is i rC^ NXX-k_x D D , o -Nxxk\x^ , . xjZ. 'Nxkx / , 0 z rkkB xN^ / k^x D D , 0 r^N^yB .xNxx^\x° , 0 , or H xN-x^;z I JI T N^|P"N JT"^ OH , and L is O 0 B
[0189] In some preferred embodiments of the present disclosure, G is . or
[0190] In some preferred embodiments of the present disclosure, G is is or is 0 o A / zo
[0191] In some preferred embodiments of the present disclosure, G is , and L , and L , and L .
[0192] In some preferred embodiments of the present disclosure, G is is
[0193] In some preferred embodiments of the present disclosure, G is is H
[0194] In some preferred embodiments of the present disclosure, G is and D or or and L is
[0195] In some preferred embodiments of the present disclosure, G is or In some preferred embodiments of the present disclosure, G is
[0196] o az xx / x xx A'nA *^xN\xxx D D A / V rk ^o^kk^N^I X^J kxN^xk^ D D 0 azxd5<d AnA n^^i k^x^k k^Ax D D 0 AZ V 1^¼ |X\^k ANXxk\xO az. v__. ^x-k Z 0 / \ N 1 k '-xk '\zNXx"'k_x° o AZ / . XX XX / ^A^^ Z ° / \ N I k^X^J kx-Nxxk\xO D D 0 A / . _ . / Al^ Z 0 N | k ^xk AN^A^0 0 AZ A^ / E k^xk \xNx / ^X° 0 A / DyDlLD 0^6 kx-NxA-xX , , o a / V a / A ^° r^'^k k^N^xk^ D D , , yB aAVX xA' z ° n । । xy kxN^Ax / ° , , O AZ x-xx^- -"X xXxkA^kB Z O kp n y < ixk A N^k dA d , , 0 3 A / °y° X 'ks ^° xkN | k^Xxk kxN^xkx-0 , , o A / k'^^kB l^xN^k\^ , , 0 AZ kA^^B ’^o'AA'^'A A A kxN^ J\ X , x^ \XXX , or . is H
[0197] In some preferred embodiments of the present disclosure, G , and L is , and L is , and L .
[0198] In some preferred embodiments of the present disclosure, G
[0199] In some preferred embodiments of the present disclosure, G is
[0200] In some preferred embodiments of the present disclosure, G is is or , and L
[0201] In some preferred embodiments of the present disclosure, G is , and L is or
[0202] In some preferred embodiments of the present disclosure, G is is or In some preferred embodiments of the present disclosure, G is and L
[0203] or
[0204] In some preferred embodiments of the present disclosure, G is and L is or
[0205] In some preferred embodiments of the present disclosure, G is and E is XX XX. XX . XX XX 1 Cl y^cD3 y n^o xna° rN^0 rNx Y" V Y" Y ° , 0 , ° , ° , or 0
[0206] In some preferred embodiments of the present disclosure, G is XX XX. XX. XX XX X Cl yA y^"cd3 y^ n^o <Nx° xNx° rNx° rNx yh Xh X X X 0 , 0 , 0 , ° , or 0
[0207] In some preferred embodiments of the present disclosure, G is H / A A nr f hi sx JX. / JX\ JOI y^° (LX J^X'y X'Y0 <n"Y° < NO (^NH j^nh L OH , and E is , ^, aO Y "cd3 ynx° YH 0 Ycd3 X JH . H X A, OH , and E is Ycd3 JH . H Xx OH or Xi XX X u a Ycd3 nY° <na° rNH YH O , O , or
[0208] In some preferred embodiments of the present disclosure, G is 0H , and E is
[0209] In some preferred embodiments of the present disclosure, G is OH , and E is
[0210] In some preferred embodiments of the present disclosure, G is and E is
[0211] In some preferred embodiments of the present disclosure, G is 0H , and E is
[0212] In some preferred embodiments of the present disclosure, G is and E is or
[0213] In some preferred embodiments of the present disclosure, G is and E is , or
[0214] In some preferred embodiments of the present disclosure, G is 0H , and E is or
[0215] In some preferred embodiments of the present disclosure, G is and E is
[0216] In some preferred embodiments of the present disclosure, G is OH , and E is or
[0217] In some preferred embodiments of the present disclosure, G is
[0218] In some preferred embodiments of the present disclosure, G is 0
[0219] In some preferred embodiments of the present disclosure, G is is .
[0220] In some preferred embodiments of the present disclosure, G is , and E is . In some preferred embodiments of the present disclosure, G is
[0221] . preferred embodiments of the present disclosure, G
[0222] In some 0
[0223] In some preferred embodiments of the present disclosure, G , and E is OH and E , and E 0 H
[0224] In some preferred embodiments of the present disclosure, G is , and E .
[0225] In some preferred embodiments of the present disclosure, G is , and E O In some preferred embodiments of the present disclosure, G
[0226] 0 preferred embodiments of the present disclosure, G
[0227] In some o preferred embodiments of the present disclosure, G
[0228] In some .
[0229] In some preferred embodiments of the present disclosure, L is
[0230] In some preferred embodiments of the present disclosure, L is and E is
[0231] In some preferred embodiments of the present disclosure, L is and E is or
[0233] In some preferred embodiments of the present disclosure, L is
[0234] In some preferred embodiments of the present disclosure, L is
[0236] In some preferred embodiments of the present disclosure, L is and E is
[0237] In some preferred embodiments of the present disclosure, L is and E is
[0238] In some preferred embodiments of the present disclosure, L is . or
[0241] In some preferred embodiments of the present disclosure, L is
[0245] In some preferred embodiments of the present disclosure, L is
[0249] In some preferred embodiments of the present disclosure, L is and E is
[0250] In some preferred embodiments of the present disclosure, L is
[0251] In some preferred embodiments of the present disclosure, L is and E is or
[0252] In some preferred embodiments of the present disclosure, L is A / OH B O
[0254] In some preferred embodiments of the present disclosure, L is or and E is or
[0257] In some preferred embodiments of the present disclosure, L is B O
[0258] In some preferred embodiments of the present disclosure, L is O' , and E is 0 or
[0259] In some preferred embodiments of the present disclosure, L is 0
[0261] In some preferred embodiments of the present disclosure, L is
[0262] In some preferred embodiments of the present disclosure, L is
[0263] In some preferred embodiments of the present disclosure, L is , L is CN
[0267] In some preferred embodiments of the present disclosure, G is A A , B B D N D N 0 A* , or D O N^B J ’B , and E is V Aa„ / N^° rNy° rNy° rNY° rNY° L / nh ><nh >rNH 0 ,0,0,0, or O . O A N , and E is 0 0 , or . preferred embodiments of the present disclosure, G
[0269] In some is OH , L is 0 a / b yyy^ 1 L-nUJ A / ) °YY HO , o A / B A / Y^x^J ’ oY^r k / NYY , o A / B A / YY^n^ Y^''Y 1 YY, / O , or ‘ Y Yd dl id / id ■ c Y^° n^o n^.o rNY° rNY° L^nh Y-nh Y<nh Y-nh 0 ,0,°,°, or
[0270] In some preferred embodiments of the present disclosure, G is o a / yy'Y / b a ^° / \ Y Y^"Y ^oY^n k Y 21 OH , 0 A / Yn'^ / B y)A t YY az kYxYxY YY^y^n - , o b n"Y , o B J,i Y^\Y , 0 i d-Y^6 N^ A J J TO • , and E is Yd YYdcd3 <N^° Y 0 . H N U N^i^N ^YnY oh , L is rY^ xYx^Y / ON , 0 yY vnAy , B In some preferred embodiments of the present disclosure, G is L is , and and E is
[0271] E is
[0272] In some preferred embodiments of the present disclosure, G is 0H , L is or and E is 0
[0273] In some preferred embodiments of the present disclosure, G is ocd3 , L , L B B B
[0274] In some preferred embodiments of the present disclosure, G is , and E is is is O B or and E is
[0275] In some preferred embodiments of the present disclosure, G is L is B is present disclosure, G is
[0276] In some preferred embodiments of the , L N ; preferably, G is . OH H N
[0277] In some preferred embodiments of the present disclosure, G is OH , and E is is is
[0278] In some preferred embodiments of the present disclosure, G is , L is , L is 0 , or , and E is
[0279] In some preferred embodiments of the present disclosure, G , and E is .
[0280] In some preferred embodiments of the present disclosure, G is , L is , and E is
[0281] In some preferred embodiments of the present disclosure, G is OH xf3 , L is 0 O B , and E is . OH preferred embodiments of the present disclosure, G is some
[0282] In or 0 azdxD ZvVA'Z A^xA A / N\Axx D D 0 A z 5<D 3<d Z^O'^y^N'^ A^xA A^N^Ax / D D 0 az dxd AA'Sa AxxA ^N^AxA a z AD 5<d xn a ,A° N^A A^xA k^N^Jx^x o A D D ZX-A / O a? V X VX ^ l^^N\Axx D D , , / B 5 * / V__Xx z 0 Ax N i A X\J A. / N\Axx , , o a / ^A'A^AA z ° Ax N i Axxx A^N\xAx D D , , o , Xx° xA / B ^°aAi r^A AnvAz D D , , 0 A / VAd X^nA / A Z< / -- / \ ^\| I ' B Arj^X' z 0 / A N i A a A ° a A^n,J\A D D , 0 A / V / x Pl^A AZ Z°Y n z A^\A z o i A^n^A\^0 , o AZ / \ z o An A A x i d d ill A Z \Z \ / nxA\ / ° z'o A' D D L , ? AZ az V zb AcOO ^°''Az''n' / A A^xA k A / N. A\ / 0 X- xXX / , or X xX Xx I z B N | AXxA A^NxJ\ / D D , o AA / AZa n i A A^n\Ax / ° , .__xA A N i A^xA A A^n^A^o , o x xA^6 \zNxJ\^O D D , and E is preferred embodiments of the present disclosure, G
[0283] In some is OH or , or , and E
[0284] In some preferred embodiments of the present disclosure, G is OH , L is and E is ,
[0285] In some preferred embodiments of the present disclosure, G is H ^■''CD3 OH , L is O A / > D D A°^Q^p 'VyV , 0 ^^QXP^6 , . H OH or 0 pN^yB 7 , o P^B i r\> -VP D D , 0 r^B -! p-P 7B ,NPJ D D , 0 A , and E is or . , or H '3
[0286] In some preferred embodiments of the present disclosure, G is L is , , , or or and E is '3
[0287] In some preferred embodiments of the present disclosure, G , L is , L is or
[0288] In some preferred embodiments of the present disclosure, G o o b AZ z O' , and E is , and E is OH , or 0H preferably, G is H OH . some preferred embodiments of the present disclosure, G is
[0291] In A , L is , and E is or , or
[0292] In some preferred embodiments of the present disclosure, G is . or B A / B , and E is
[0293] In some preferred embodiments of the present disclosure, G or B A /
[0294] In some preferred embodiments of the present disclosure, G or a? / ""O' NH or
[0295] In some preferred embodiments of the present disclosure, A in L is linked to G.
[0296] In some preferred embodiments of the present disclosure, W is CRza.
[0297] In some preferred embodiments of the present disclosure, Rza is H or halogen, e.g., halogen.
[0298] In some preferred embodiments of the present disclosure, Z is N.
[0299] In some preferred embodiments of the present disclosure, Rzd is methyl, deuterated methyl, halomethyl, -CD2F, -CDF2, or C2-6 alkyl, wherein the C2-6 alkyl is optionally substituted with 1 or more hydroxy, deuterium, or halogen; preferably, Rzd is methyl or deuterated methyl.
[0300] In some preferred embodiments of the present disclosure, n is 0.
[0301] In some preferred embodiments of the present disclosure, Rzc is phenyl, naphthyl, or benzo[b]thiophene, wherein the phenyl, naphthyl, or benzo[b]thiophene is optionally substituted with 1 or more R1'.
[0302] In some preferred embodiments of the present disclosure, in the compound of formula I:
[0303] G is ;
[0304] W is CRza;
[0305] Rza is hydrogen or halogen;
[0306] Z is N;
[0307] Rzc is phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole, or pyridine, wherein the phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole, or pyridine is optionally substituted with 1 or more R1';
[0308] each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, C2-6 alkenyl, C2-6 alkynyl, C2-6 deuterated alkynyl, or cycloalkyl (preferably, the cycloalkyl is C3-6 cycloalkyl, e.g., cyclopropyl);
[0309] R21 is hydrogen;
[0310] R22 is hydrogen;
[0311] Rzd is hydrogen, deuterium, methyl, deuterated methyl, halomethyl, -CD2F, -CDF2, or C2-6 alkyl, wherein the C2-6 alkyl is optionally substituted with 1 or more hydroxy, deuterium, or halogen;
[0312] R3a is hydrogen, halogen, hydroxy, cyano, C1-3 alkyl, C1-3 deuterated alkyl, or C1-3 haloalkyl;
[0313] n is 0 or 1;
[0314] , and L is linked to G via A;
[0315] 0 Cl 0 .
[0316] In some preferred embodiments of the present disclosure, in the compound of formula I:
[0317] G is
[0318] W is CRza;
[0319] Rza is halogen;
[0320] Z is N;
[0321] Rzc is phenyl, naphthyl, or benzo[b]thiophene, wherein the phenyl, naphthyl, or benzo[b]thiophene is optionally substituted with 1 or more R1';
[0322] each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, C2-6 alkenyl, C2-6 alkynyl, C2-6 deuterated alkynyl, or cycloalkyl;
[0323] R21 is hydrogen;
[0324] R22 is hydrogen;
[0325] Rzd is methyl or deuterated methyl;
[0326] n is 0; In some preferred embodiments of the present disclosure, G2 is G2-1, G2-2, G2-3, or G2-4:
[0329] B (R3a) R G2-1 G2-2 G2-3 G2-4
[0330] wherein Rzc, Rzd, R3a, and n in G2-1, G2-2, G2-3, or G2-4 are as described and defined in the present disclosure.
[0331] In some preferred embodiments of the present disclosure, G2 is G2-2-1, G2-2-2, or G2-2-3:
[0332] wherein Rza, Rzb, Rzc, Rzd, R3a, and n in G2-1, G2-2, G2-3, or G2-4 are as described and defined in the present disclosure. In some embodiments of the present disclosure,
[0333] , , , , N^y^N N^y^N yV^nA / ’tV^^ Vy^n^ F
[0334] , F , or F • In some preferred embodiments of the present disclosure, G2 is: ^^ci aX nh2 F V Fi cy NH2 O hA jl F O N" F H ^N' SA '''A xfA H N '"nA N N A H N H ^-N JI AA V N ''f ' N AaA a At N \A F nh2 H ^Ns O N'" cf3 nAA.-n jVz nh2 H T UI 0 C A n^A'n H -N A / NH2 NH2 , , H H . NN Cl NH2 A J A N I jl C f D3C 0 cf3 CjCX nh2 D3Q H A 'lA H '"N^ A"' Cl Mx! cA NH2 H D3A—CJ ui A ? N ^r 'N AaA a pj N NH2 H D3S-CN' O "A cf3 nAA-n JAA z OH H AA Aa (£A jA nh2 D3C I O A N ' I jl A F NH2 H N N^ N ' N^y H O X NH2 h2n CN ' AA N svAA Xj F^\^ H ^N Q X AA । 1 A y n F clXi" NH2 , d OH H2N S J F^ H NH2 CN H n^V^n aC a Y N F nh2 h2n CN ' r=( n CCa AT F^ / d Y AV : jl J A F OH , H / rN 0 lAyA AC A / n 7 F , H / TN A An Xa , H N H X I °\ - 'hXl SF5 nX^N “ly / X “^Xy / x nh2 oh , H o 'X cf3 nX^n ciXUa YX^ or OH .
[0335] In some preferred embodiments of the H _-N H \ X, / XX \_XX o u .. 0 'X CF3 nX^N h2n on \ m i xl A X( N N sx X X X X / UJI YYj N 7 V F-W F , OH , ,
[0336] In some preferred embodiments of the H H DX N; \ N> X X V nX\ V XXn C|\^ ciXAXX H 1 N 7 li jT 1 N 7 xj f YY f OH NH2 , , H H N o "X SF5 N XX N Y 7 ci^YYXxmX^ ciXlXklX z r|O 7 Yx^> XX F AA f nh2 oh , ,, H X X CF3 nX^N “Y^x7 present disclosure, G2 is: NH2 , H X Y nX^n 3 r OH . H D3C\Xx) 0 X Y nX^n ci^XYxAnX / present disclosure, G2 is: NH2 , H H < rN\ ,--N \ X > —1>Y 0 'N FVF n°Xn YX 1 c'Yxx “yxx nh2 , or nh2 , e.g.,
[0337] In some preferred embodiments of the present disclosure, G2 is: H XN V H N 'N ' N N^y , H N H rA o^'> NAy^N AA A / Y N 7 F H XN O y nAAn YA H XN . > NH2 NH2 , , H H N nh2 XF| NH2 O A' F O N" jl F NH2 cf3 “J NH2 V'.”. X nh2 F J I JI A I yl "O^F D3C o H A 'A AN "A" H 1A Clyr N nh2 H D3Q / TN yy-, y y-, > yy, nh2 H D3C\JXJ I O^'N^ Y n^t^n 1 Jj y y< n y f NH2 , H2N s. F' NH2 , H .--N q CN 1 H2N nXX ; . JO y JO, s. N z X F F- NH2 D3Q O CN \ N jl F H iTN 'IX "A , or , h2n s. F' dyA UnyX YA nh2 , H d3c_yNj JI A A' xx n A^A AaA A / yr nh2 , H d3c_yN\ cf3 nAAs,n AAA r f nh2 , H nh2 , H r-N - CN k A N Y N . Al A / y n 7 F
[0338] In some preferred embodiments of the present disclosure, G2 is: H H H H \ A \ \ A \ N N \ I X I ' N NN • ' '• > nh2 nh2 nh2 nh2 ,, , , H H H yxy h mx NH2 F NH2 NH2 , , ,, H H H < / XN\ DaQ yN\ d3c |l| 0 N 0 N 0 N yM ^x nh2 nh2 nh2 , ,, H nh2 , H H 3 MTJ D3ChTJ y$x ^4x yy 7 yx NH2 , NH2 , NH2 , NH2 , or OH
[0341] In some preferred embodiments of the present disclosure, G2 is: OH and L is
[0342] In some preferred embodiments of the present disclosure, G2 is or
[0343] In some preferred embodiments of the present disclosure, G2 is NHz , L is In some preferred embodiments of the present disclosure, G2 is L is
[0344] or iTl t O ^y^CI A / N r / N \ / >0 px 7 r Y r A \nh 1 D D , and E is 0 , id iCl y yd3 xNy° YH YH $ , or .
[0346] In some preferred embodiments of the present disclosure, G2 is 0 a z x\ b az z 0 N > [ '\X Z 0 N , o A / B A / k^N , _. JY A / V rO^B \^Nx , 0 A / Z^N^ / b z''o / ''y^''n'^ r^x^ ^X / N\x^x / , 9 A / DvD a z xD 3<d r^N "z b z^°'xy^'N' / ^ Z^x^J k^N^X^ / , nO YBCD3 xN^-0 ^Y° ynh kNH 0 0 , , H r'N o h2n cn x Z XV NXf 'N S. V Jz X / fUTn^ . F , L is 0 x^O^6 'Jx^xk^ , o r^N^kj / B ^Xx / O , 0 rk^1^6 , o Nxxk^ D^ D , i rO^76 N\xk^x D D , , and E is l! Iki Y'cd3 / nY° r^x0 YH \NH 0 , or 0 .
[0347] In some preferred embodiments of the present disclosure, G2 is 0 az YYa / b a / z o y N । r xY * o Y n ^N\YaY V / , o A / B A / ■ o^y^'n YXX X^Xm^ Y^n'X\X Y^r , ?, A / V Vzx rO^B A° iX X-N-X^sX , 0 A / YD Y^^ / b x^y^^X Y^xY Y Y / N\Yxx , 9 A / DvD A Z XD X° PN ? B YY^XlX Xo-x^n^y Y^\Y y Y / n\Yx / , n J A / V az ^Yx x\YY^Yb YqYYYi^ax zO r N Y J 1 1 I xNY\J^ Yx D D , H D3Y-Y.y h2n CN Y X ' X / N ;-f ;'N S X Y X 7 pljf . F , L is 0 ySy^6 MxX'Xa' , o Y''N^''V b Y-. / O , xd^76 , o X-Cj J^B 'n\X^^ D D , 0 'i xX^6 -N^Xx^ / D D , ......X JvY / D D , o AZ V - .....1 , o D D , o ^-¾ D D , 0 D D , 0 A / D\ / D Ao^An^ pp Zb , 0 AZ V ^N^Vb A / , o A / f^'N^y’e ^CijX D D , and E is l! il! / Ny° r^y0 YH VH $ , or . O ^¾^7, , ..... , 0 A / V o / Ve ' ” -.....OX J D D , AA0>y,p pO^8 , o A / r^N^ / B Z ° N | I D D , 0 '^XPJS'‘ , or "Vl / yXl J OCDa Y"^0 ^Ny° ^'T0 r^1^0 Lp IpH L NH 0 0 o , ,,
[0348] In some preferred embodiments of the present disclosure, G2 is 0 a? z 0 N y | Z o / ( N 'x^N'XXX , o A / r^'N^yB A / Zx , ? A / V X'Y'rO Y"^ AXA \Xxx-o a / ^' , 0 A / (^^^( / b ^X / N-XXA , 9 A / V A Z XD YD Y^n "y B Y'y'Y ^°'i / ^N' / xy Y^xY Y Y / N\Yxx , n A / V / □ az f^yA / B YqYYYn-^x f o Y n Y YJ i । 1 ^>Jx^^ D D , o a? xD r^B az vd ^o^yY'N^Y r^x^ n^ k^N^Yx / J , o AA / \ X\r^V^B A D D D ^° / \ N i r^xxJ A z x J<D L^n^JxY ao'yyn^ D D l"X / - , H XN ZN^ h2\ CN X / xy n-x'n s v JL x\ y > ijf N^ . F , L is 0 1 xO^7" NxXxX , o Y''n^Y b i^x --, / ° , ^pX , o N\XYx D D , 0 XxYb -Nx^Yx D D , .....X. JxYx^ D D , 0 y r^O^8 -NxXXX , , rY , , and E is 0 0 , or .
[0349] In some preferred embodiments of the present disclosure, G2 is oh , L is 0 , , 0 0 , or . and E is H
[0350] In some preferred embodiments of the present disclosure, G2 is OH , L is , and E is G2 OH is , L is preferred embodiments of the present disclosure,
[0351] In some or
[0352] In some preferred embodiments of the present disclosure, G2 is 0H , L is 0 AZ ' 9 / CJh C D D , and E is 0 , N^° rV YH YH $ , or .
[0353] In some preferred embodiments of the present disclosure, G2 is 0 az ~ yx^^yB A? z o yy n y y x^J ^o y n y y^N\ / \x k^N , o A / B A / ■^o^y^^N^y yy^y ^o^y^^N^y k^N.xy^o y^-N- , ? A z V v-o-xx xO^B , 0 A? \ / D y^N^yB y^xx^ y^'x-yx , 9 A / DVD A Z XD 5<d f^N / 7 b ^cry^N^ Z^o'xy / ^N'^ y^xx* \ / y^N\x\x , Y^ocd3 y^o Ny° r yh 0 0 , , H r N cFy& OH , L is 0 , o y^'N^y b , jypN^B , o J\X\^ D D , i Hy^6 Nxx\^ D D , . H OH preferred embodiments of the present disclosure, G2 is , L is
[0354] In some , and E is , , OH L is
[0355] In some preferred embodiments of the present disclosure, G2 is and E is
[0356] In some preferred embodiments of the present disclosure, Rzc is: 7' OH OH OH OH OH OH OH , , , , ,, i^a i A OH OH OH OH OH OH ,, ,, ,,, ci. 67 OH OH OH OH ,,,, , , . . . ? . . ' C- ■ ' ,,,,,,, OH 2> Sr 'Vc CS ,,, ,,, ,,, ,,, the following compounds: OH P26413667AU OH P26413667AU 00 P26413667AU H OMe O H F 1 7'7- ,.... 7; OH D D < O H / N CD3 F y JC^YTOypCj-oh d d r Y ¥ 0 H F hi N 1 JI 1 ° fi CTi YD W' °vyy y. OH D 0 7 O H <YrCD3 I J I ° Cl ClO dC uYN Yo Y OH D D H FN 1 III 7 o (XyJOV-y-p Y F N 1 OH D D r O F Y o OH D D ( Y 0 H yyo3 F III o 7 JyC1^o^1y 1 OH fl NH L.NH O , H / N^CD3 F s? r JC^o^OyC^ OH D D ( Y 1 YH 0 , F Y o k I OH D D ( T nh ^>rNH 0 , H F N G 7 7-7^7-7^ 1 OH | T rNH YH 0 0 , F || Y o 7OMe *J?^YVuCpi9r J OH ( V NH YYNH 0 , H Yx003 F III 0 HllY °Me TJ T j .N j J Y0Me 0 xnyy° OH 1 । 1 VH 0 of the following structures: P26413667AU H H
[0364] the compound that elutes first between under the following chromatographic conditions: chromatographic column: CHIRALPAK IC(IC00CD-TB016), 0.46 cm x 15 cm I.D., 10 mm mobile phase: MeOH / ACN (acetonitrile) / DEA (diethanolamine); gradient: MeOH / ACN / DEA = 65 / 35 / 0.2 (V / V / V); flow rate: 1 mL / min; column temperature: 35 °C; preferably, the retention time of the compound that elutes first is 6.913 min; or the compound that elutes later between
[0365] H H OH O and 0 under the following chromatographic conditions: chromatographic column: CHIRALPAK IC(IC00CD-TB016), 0.46 cm x 15 cm I.D., 10 ^m; mobile phase: MeOH / ACN / DEA; gradient: MeOH / ACN / DEA = 65 / 35 / 0.2 (V / V / V); flow rate: 1 mL / min; column temperature: 35 °C; preferably, the retention time of the compound that elutes later is 9.062 min. In some preferred embodiments of the present disclosure, the configuration of
[0366] in is the same as the configuration of the compound that elutes first between and Boc Cl under the following chromatographic conditions: column: Daicel chiralpak AD_3, 3 x 150 mm, 3 um; mobile phase A: CO2; mobile phase B: MeOH (0.1% diethanolamine); gradient: mobile phase A / mobile phase B = 50 / 50 (V / V), elution time: 6 min; flow rate: 1.5 mL / min; wavelength: 214 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes first is 2.248 min. In some preferred embodiments of the present disclosure, the configuration of F in
[0367] that elutes later between and is the same as the configuration of the compound under the following chromatographic conditions: column: Daicel chiralpak AD_3, 3 x 150 mm, 3 ^m; mobile phase A: CO2; mobile phase B: MeOH (0.1% diethanolamine); gradient: mobile phase A / mobile phase B = 50 / 50 (V / V), elution time: 6 min; flow rate: 1.5 mL / min; wavelength: 214 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes later is 2.350 min. disclosure, the configuration of In some preferred embodiments of the present
[0368] is the same as the configuration of the compound that elutes first between 3, 3 x 150 mm, 3 um; mobile _ under the following chromatographic conditions: column: Daicel chiralpak AD phase A: CO2; mobile phase B: MeOH (0.1% diethanolamine); gradient: A / B = 80 / 20 (V / V), elution time: 6 min; flow rate: 2 mL / min; wavelength: 214 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes first is 1.408 min.
[0369] In some preferred embodiments of the present disclosure, the configuration of -J— or in is the same as the configuration of the compound that Boc Boc elutes first between Trt N 1 and Trt under the following chromatographic conditions: column: chiralpak-IG-3, 150 mm x 3 mm, 3 um; mobile phase A: CO2; mobile phase B: MeOH (0.1% diethanolamine); gradient: A / B = 90 / 10 (V / V), elution time: 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm or 254 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes first is 1.357 min.
[0370] In some preferred embodiments of the present disclosure, the configuration of in is the the configuration of the compound that elutes later between same as and Boc 1 n^cd3 Cl Cl under the following chromatographic conditions: column: Daicel CHIRALPAK IB- 3, 3 x 150 mm, 3 um; mobile phase A: CO2; mobile phase B: MeOH; gradient: A / B = 70 / 30 (V / V), elution time: 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm or 254 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes later is 1.681 min.
[0371] In some preferred embodiments of the present disclosure, the configuration of in is the same as the configuration of the compound that elutes first between and under the following chromatographic conditions: column: Daicel CHIRALPAK IB-3, 3 x 150 mm, 3 ^m; mobile phase A: CO2; mobile phase B: MeOH (0.1% diethanolamine); gradient: A / B = 70 / 30 (V / V), elution time: 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes first is 2.513 min. In some preferred embodiments of the present disclosure, the configuration of
[0372] is the same as the configuration of the compound that elutes first between and under the following chromatographic conditions: column: Daicel CHIRALCEL OD-3, 150 mm x 3 mm, 3 um; mobile phase A: CO2; mobile phase B: MeOH (0.1% diethanolamine); gradient: A / B = 60 / 40 (V / V), elution time: 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes first is 1.693 min;
[0373] or the configuration of same as the configuration of the compound that elutes later between under the following chromatographic conditions: column: Daicel CHIRALCEL OD- 3, 150 mm x 3 mm, 3 nm; mobile phase A: CO2; mobile phase B: MeOH (0.1% diethanolamine); gradient: A / B = 60 / 40 (V / V), elution time: 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C; preferably, the retention time of the compound that elutes later is 1.942 min.
[0374] In some preferred embodiments of the present disclosure, the prodrug of the compound of formula I is as follows: O OMe O O OMe O H2N S F OMe O or
[0375] The present disclosure provides a pharmaceutical composition, comprising a therapeutically effective amount of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.
[0376] The present disclosure provides a method for degrading the KRAS G12D protein, comprising contacting the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof with the KRAS G12D protein.
[0377] The present disclosure provides use of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof as a medicament for treating or preventing a KRAS G12D-mediated disease or disorder.
[0378] The present disclosure provides use of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof as a medicament for treating or preventing a KRAS mutation-mediated disease or disorder.
[0379] The present disclosure provides use of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof in the preparation of a medicament for treating or preventing a KRAS mutation-mediated disease or disorder, e.g., a KRAS G12D mutation-mediated disease or disorder.
[0380] The present disclosure provides use of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof in the preparation of a medicament for treating or preventing a disease or disorder (cancer) caused by the interaction between KRAS G12D and the SOS1 or SHP2 protein.
[0381] The present disclosure provides use of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof in the preparation of a medicament for treating or preventing a cancer.
[0382] The present disclosure provides use of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof in the preparation of a medicament for treating or preventing pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial cancer, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer, and sarcoma (e.g., the disease described above is a disease or disorder caused by the interaction between KRAS G12D and the SOS1 or SHP2 protein).
[0383] The present disclosure provides a method for treating or preventing a KRAS G12D-mediated disease or disorder, comprising administering to a patient in need thereof a therapeutically effective amount of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof.
[0384] The present disclosure provides a method for treating or preventing a disease or disorder (e.g., cancer) regulated by the interaction between KRAS G12D and the SOS1 or SHP2 protein, comprising administering to a patient in need thereof a therapeutically effective amount of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof.
[0385] In certain embodiments of the present disclosure, the cancer is selected from:
[0386] heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, and liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma;
[0387] lung: bronchial cancer (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, and adenocarcinoma), alveolar carcinoma (bronchioloalveolar carcinoma), bronchial adenoma, sarcoma, lymphoma, chondroma, hamartoma, and mesothelioma;
[0388] gastrointestinal tract: esophagus cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, and lymphoma), stomach (cancer, lymphoma, and leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, and vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi’s sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, and fibroma), and large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, and leiomyoma);
[0389] genitourinary system: kidney (adenocarcinoma, embryonal carcinosarcoma (Wilms’ tumor), lymphoma, and leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, and adenocarcinoma), prostate (adenocarcinoma and sarcoma), and testis (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, and lipoma);
[0390] liver: liver cancer (hepatocellular carcinoma), intrahepatic cholangiocarcinoma, hepatoblastoma, malignant hemangioendothelioma, hepatocellular adenoma, and hemangioma;
[0391] biliary tract: cholangiocarcinoma, gallbladder cancer, ampullary cancer, and intrahepatic cholangiocarcinoma;
[0392] bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing’s sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, chondroma (osteocartilaginous exostosis), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor;
[0393] nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, and Paget’s disease of bone), meninges (meningioma, meningeal sarcoma, and glioma), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinal cell tumor (pinealoma), glioblastoma multiforme, oligodendroglioma, neurilemmoma, eye cancer, and congenital tumor), spinal neurofibroma, meningioma, glioma, and sarcoma);
[0394] gynecology and obstetrics: uterus (endometrial cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, and unclassified cancer), granulosa-theca cell tumor, ovarian Sertoli-Leydig cell tumor, dysgerminoma, and malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, and melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botulinum sarcoma (embryonal rhabdomyosarcoma), and fallopian tube (cancer);
[0395] hematology: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disease, multiple myeloma, and myelodysplastic syndrome), Hodgkin’s disease, and non-Hodgkin’s lymphoma (malignant lymphoma);
[0396] skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi’s sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, and psoriasis; and
[0397] adrenal gland: neuroblastoma.
[0398] In certain embodiments of the present disclosure, the cancer is a KRAS G12D-associated cancer.
[0399] In certain embodiments of the present disclosure, the cancer is non-small cell lung cancer, small cell lung cancer, colorectal cancer, rectal cancer, or pancreatic cancer.
[0400] The present disclosure further provides the compound of formula I, or the stereoisomer thereof, the atropisomer thereof, the pharmaceutically acceptable salt thereof, the pharmaceutically acceptable salt of the stereoisomer thereof, or the pharmaceutically acceptable salt of the atropisomer thereof described above for use in the treatment of a disease or disorder associated with the KRAS G12D mutant protein.
[0401] The present disclosure further provides the compound of formula I described above, wherein the disease or disorder associated with the KRAS G12D mutant protein is a cancer associated with the KRAS G12D mutant protein.
[0402] In certain embodiments of the present disclosure, the cancer is selected from pancreatic cancer, colorectal cancer, endometrial cancer, and lung cancer.
[0403] In certain embodiments of the present disclosure, the lung cancer is selected from non-small cell lung cancer and small cell lung cancer.
[0404] In certain embodiments of the present disclosure, the cancer and the KRAS mutation-mediated disease or disorder are independently selected from gastric cancer, pancreatic cancer, pancreatic adenocarcinoma, and colon cancer.
[0405] Detailed description: Unless otherwise stated, the following terms used in the specification and claims have the following meanings.
[0406] “Alkyl” refers to a saturated aliphatic hydrocarbon group, including linear or branched alkyl; for example, C1-C8 alkyl refers to an alkyl group containing 1-8 carbon atoms, and the C1-C8 alkyl includes C1-C2, C1-C3, C1-C4, C1-C5, C2-C3, or C2-C4 alkyl, etc., such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, or various branched isomers thereof, preferably C1-C6 alkyl, and more preferably C1-C4 alkyl. The alkyl may be substituted or unsubstituted. In some embodiments, the alkyl is C1, C2, C3, C4, C5, C6, C7, or C8 alkyl.
[0407] “C1-3 alkyl” refers to a linear or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C1-3 alkyl includes C1-2 alkyl, C2-3 alkyl, etc., which may be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methyne). Examples of C1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.
[0408] “Heteroalkyl” refers to a saturated aliphatic hydrocarbon group in which a methylene group (-CH2-) is replaced by a heteroatom (e.g., O, S, or N), a heteroatom group (e.g., -C(O)-, -S(O)-, -S(O)2-, -C(O)O-, or -OC(O)-), C(O)NH-, -NHC(O)-, vinylene, or ethynylene, including linear or branched heteroalkyl; C1-C8 heteroalkyl refers to an alkyl group containing 1-8 carbon atoms in which at least one methylene group is replaced by a heteroatom or a heteroatom group, such as -C(O)-CH3, -CH2-O-CH3, -CH2-S-CH3, -CH2-S(O)-CH3, -CH2-S(O)2-CH3, -CH2-S-CH2-CH3, -CH2-O-CH2-CH3, -CH2-O-CH2-CH3, -CH2-S(O)-CH2-CH3, or various branched isomers thereof, preferably C1-C6 heteroalkyl, and more preferably C1-C4 heteroalkyl. The heteroalkyl may be substituted or unsubstituted. In some embodiments, the heteroalkyl is C1, C2, C3, C4, C5, C6, C7, or C8 heteroalkyl.
[0409] “Alkenyl” refers to an alkyl group as defined above that consists of at least two carbon atoms and at least one carbon-carbon double bond; “C2-8 alkenyl” refers to a linear or branched alkenyl group containing 2-8 carbon atoms, including but not limited to vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, etc., preferably “C2-6 alkenyl”, and more preferably “C2-4 alkenyl”. The alkenyl may be substituted or unsubstituted. In some embodiments, the alkenyl is C2, C3, C4, C5, C6, C7, or C8 alkenyl.
[0410] “C2-4 alkenyl” refers to a linear or branched hydrocarbon group consisting of 2 to 4 carbon atoms and containing at least one carbon-carbon double bond. The carbon-carbon double bond may be located at any position of the group. The C2-4 alkenyl includes C4 alkenyl, C3 alkenyl, and C2 alkenyl; the C2-4 alkenyl may be monovalent, divalent, or polyvalent. Examples of C2-4 alkenyl include, but are not limited to, vinyl, propenyl, butenyl (including 1-, 2-, or 3-butenyl), etc.
[0411] “Alkynyl” refers to an alkyl group as defined above that consists of at least two carbon atoms and at least one carbon-carbon triple bond; “C2-8 alkynyl” refers to a linear or branched alkynyl group containing 28 carbon atoms, including but not limited to ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, preferably “C2-6 alkynyl”, and more preferably “C2-4 alkynyl”. The alkynyl may be substituted or unsubstituted. In some embodiments, the alkynyl is C2, C3, C4, C5, C6, C7, or C8 alkynyl.
[0412] “C2-4 alkynyl” refers to a linear or branched hydrocarbon group consisting of 2 to 4 carbon atoms and containing at least one carbon-carbon triple bond. The carbon-carbon triple bond may be located at any position of the group. The C2-4 alkynyl includes C4 alkynyl, C3 alkynyl, and C2 alkynyl; the C2-4 alkynyl may be monovalent, divalent, or polyvalent. Examples of C2-4 alkynyl include, but are not limited to, ethynyl, propynyl, butynyl (including 1-, 2-, or 3-butynyl), etc.
[0413] “-ylene” refers to a divalent group. For example, alkylene refers to a divalent alkyl group; alkenylene refers to a divalent alkenyl group; alkynylene refers to a divalent alkynyl group; cycloalkylene refers to a divalent cycloalkyl group; heterocycloalkylene refers to a divalent heterocycloalkyl group; arylene refers to a divalent aryl group; heteroarylene refers to a divalent heteroaryl group; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are as defined above; the “-ylene” groups may be optionally substituted or unsubstituted.
[0414] “Haloalkyl” refers to an alkyl group optionally substituted with one or more fluorine, chlorine, bromine, or iodine atoms, wherein the alkyl group is as defined above; non-limiting examples include difluoromethyl, dichloromethyl, dibromomethyl, trifluoromethyl, trichloromethyl, tribromomethyl, etc.
[0415] “Hydroxyalkyl” refers to an alkyl group optionally substituted with one or more OH, wherein the alkyl group is as defined above; non-limiting examples include hydroxymethyl, hydroxyethyl, hydroxypropyl, and hydroxyisopropyl.
[0416] “Alkoxy” refers to -O-alkyl, wherein the alkyl is as defined above; non-limiting examples include methoxy, ethoxy, isopropoxy, tert-butoxy, etc.
[0417] “Heteroalkyl” refers to a saturated aliphatic hydrocarbon group in which a methylene group (-CH2-) is replaced by a heteroatom (e.g., O, S, or N), a heteroatom group (e.g., -C(O)-, -S(O)-, -S(O)2-, -C(O)O-, or -OC(O)-), C(O)NH-, -NHC(O)-, vinylene, or ethynylene, including linear or branched heteroalkyl; C1-C8 heteroalkyl refers to an alkyl group containing 1-8 carbon atoms in which at least one methylene group is replaced by a heteroatom or a heteroatom group, such as -C(O)-CH3, -CH2-O-CH3, -CH2-S-CH3, -CH2-S(O)-CH3, -CH2-S(O)2-CH3, -CH2-S-CH2-CH3, -CH2-O-CH2-CH3, -CH2-O-CH2-CH3, -CH2-S(O)-CH2-CH3, or various branched isomers thereof, preferably C1-C6 heteroalkyl, and more preferably C1-C4 heteroalkyl. The heteroalkyl may be substituted or unsubstituted. In some embodiments, the heteroalkyl is C1, C2, C3, C4, C5, C6, C7, or C8 heteroalkyl.
[0418] “Cycloalkyl” refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent; “C3-C11 cycloalkyl” refers to a cycloalkyl group containing 3 to 11 carbon atoms; “C3-C8 cycloalkyl” refers to a cycloalkyl group containing 3 to 8 carbon atoms; “C5-C10 cycloalkyl” refers to a cycloalkyl group containing 5 to 10 carbon atoms.
[0419] Non-limiting examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc., preferably cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, preferably C3-C8 cycloalkyl, and more preferably C3-C6 cycloalkyl.
[0420] Polycyclic cycloalkyl includes spiro, fused, and bridged cycloalkyl. “Spirocycloalkyl” refers to a polycyclic group in which monocycles share one carbon atom (referred to as a spiro atom); they may contain one or more double bonds, but none of the rings has a fully conjugated n-electron system. According to the number of spiro atoms shared between rings, the spirocycloalkyl may include monospirocycloalkyl, dispirocycloalkyl, or polyspirocycloalkyl, and is preferably 7- to 12-membered dispirocycloalkyl. Nonlimiting examples of the spirocycloalkyl include:
[0422] “Fused cycloalkyl” refers to an all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with another ring in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated n-electron system. According to the number of constituent rings, the fused cycloalkyl may include bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl, and is preferably bicyclic fused cycloalkyl. Non-limiting examples of the fused cycloalkyl include:
[0424] “Bridged cycloalkyl” refers to an all-carbon polycyclic group in which any two rings share two carbon atoms not directly connected; they may contain one or more double bonds, but none of the rings has a fully conjugated n-electron system. According to the number of constituent rings, the bridged cycloalkyl may include bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl. Non-limiting examples of the bridged cycloalkyl include:
[0426] The cycloalkyl ring may be fused to an aryl, heteroaryl, or heterocycloalkyl ring, wherein the ring attached to the parent structure is the cycloalkyl; non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc. The cycloalkyl may be optionally substituted or unsubstituted.
[0427] In some embodiments, the cycloalkyl is C3, C4, C5, C6, C7, C8, C9, C10, C11, or C12 monocyclic or polycyclic (e.g., spiro, fused, or bridged) cycloalkyl.
[0428] “Cycloalkylene” refers to a divalent group obtained by removing one additional hydrogen atom from the corresponding “cycloalkyl”.
[0429] “Heterocycloalkyl” refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein one or more (e.g., 2, 3, 4, or 5) of the ring atoms are selected from nitrogen, oxygen, and S(O)r (wherein r is an integer of 0, 1, or 2), but a ring portion of -O-O-, -O-S-, or -S-S- is not included, and the remaining ring atoms are carbon atoms. “3- to 11-membered heterocycloalkyl” refers to a cyclic group containing 3 to 11 ring atoms, “5- to 10-membered heterocycloalkyl” refers to a cyclic group containing 5 to 10 ring atoms, and “3- to 8-membered heterocycloalkyl” refers to a cyclic group containing 3 to 8 ring atoms. “3- to 11-membered heterocycloalkyl” containing 1-2 heteroatoms selected from N, O, and S is preferred, and 3- to 11-membered heterocycloalkyl containing 1 or 2 N atoms is more preferred.
[0430] Monocyclic heterocycloalkyl is preferably 3- to 8-membered monocyclic heterocycloalkyl containing 1-2 N heteroatoms; non-limiting examples of the monocyclic heterocycloalkyl include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably piperidinyl and piperazinyl.
[0431] The polycyclic heterocycloalkyl includes spiro, fused, and bridged heterocycloalkyl. “Spiroheterocycloalkyl” refers to a polycyclic heterocycloalkyl group in which monocycles share one atom (referred to as a spiro atom), wherein one or more of the ring atoms are selected from nitrogen, oxygen, and S(O)r (wherein r is an integer of 0, 1, or 2), and the remaining ring atoms are carbon atoms. They may contain one or more double bonds, but none of the rings has a fully conjugated n-electron system. According to the number of spiro atoms shared between rings, the spirocycloalkyl may include monospiroheterocycloalkyl, dispiroheterocycloalkyl, or polyspiroheterocycloalkyl, and is preferably saturated “3- to 11-membered bispiroheterocycloalkyl” containing 1-2 heteroatoms selected from N, O, and S, and more preferably saturated “7- to 11-membered dispiroheterocycloalkyl” containing 1 or 2 N atoms. Non-limiting examples of the spiroheterocycloalkyl include:
[0432] H , etc.
[0433] “Fused heterocycloalkyl” refers to a polycyclic heterocycloalkyl group in which each ring in the system shares a pair of adjacent atoms with another ring in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated n-electron system, wherein one or more of the ring atoms are selected from nitrogen, oxygen, and S(O)r (wherein r is an integer of 0, 1, or 2), and the remaining ring atoms are carbon atoms. According to the number of constituent rings, the fused heterocycloalkyl may include bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocycloalkyl, and is preferably “3- to 11-membered bicyclic fused heterocycloalkyl” containing 1-3 heteroatoms selected from N, O, and S, and more preferably saturated “3- to 11-membered bicyclic fused heterocycloalkyl” containing 1 or 2 N atoms. Non-limiting examples of the fused heterocycloalkyl include:
[0434] N H , etc.
[0435] “Bridged heterocycloalkyl” refers to a polycyclic heterocycloalkyl group in which any two rings share two atoms not directly connected, and they may contain one or more double bonds, but none of the rings has a fully conjugated n-electron system, wherein one or more of the ring atoms are selected from nitrogen, oxygen, and S(O)r (wherein r is an integer of 0, 1, or 2), and the remaining ring atoms are carbon atoms. According to the number of constituent rings, the bridged heterocycloalkyl may include bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl; non-limiting examples of the bridged heterocycloalkyl include:
[0436] etc.
[0437] The heterocycloalkyl ring may be fused to an aryl, heteroaryl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heterocycloalkyl; non-limiting examples include:
[0438] , etc.; the heterocycloalkyl may be optionally substituted or unsubstituted. In some embodiments, the heterocycloalkyl is 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered monocyclic or polycyclic (e.g., spiro, fused, or bridged) heterocycloalkyl, wherein the number of heteroatoms may be 1, 2, 3, 4, or 5, and each heteroatom is independently nitrogen, oxygen, or S(O)r (wherein r is an integer of 0, 1, or 2).
[0439] “Heterocycloalkylene” refers to a divalent group obtained by removing one additional hydrogen atom from the corresponding “heterocycloalkyl”.
[0440] “Aryl” refers to an all-carbon monocyclic or fused polycyclic (i.e., rings sharing pairs of adjacent carbon atoms) group having a conjugated n-electron system; “6- to 10-membered aryl” refers to an all-carbon aryl group containing 6-10 carbon atoms, such as phenyl and naphthyl, preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocycloalkyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring; non-limiting examples include: embodiments, the aryl is 6- to 10-membered aryl.
[0442] “Heteroaryl” refers to a heteroaromatic system containing 1 to 4 heteroatoms, wherein the heteroatoms include nitrogen, oxygen, or S(O)r (wherein r is an integer of 0, 1, or 2); 5- to 6-membered heteroaryl refers to a heteroaromatic system containing 5-6 ring atoms, and 5- to 10-membered heteroaryl refers to a heteroaromatic system containing 5-10 ring atoms, preferably 5- to 6-membered heteroaryl, and more preferably 5- to 6-membered heteroaryl containing 1 or 2 N atoms; non-limiting examples include furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyrazole, imidazolyl, triazolyl, tetrazolyl, etc., preferably pyridyl. The heteroaryl ring may be fused to an aryl, heterocycloalkyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring; non-limiting examples include:
[0443] optionally substituted or unsubstituted. In some embodiments, the heteroaryl is 5-, 6-, 7-, 8-, 9-, or 10membered heteroaryl, wherein the number of heteroatoms may be 1, 2, 3, 4, or 5, and each heteroatom is independently nitrogen, oxygen, or S.
[0444] “Alkenyl” refers to an alkyl group as defined above that consists of at least two carbon atoms and at least one carbon-carbon double bond; “C2-8 alkenyl” refers to a linear or branched alkenyl group containing 2-8 carbon atoms, including but not limited to vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, etc., preferably “C2-6 alkenyl”, and more preferably “C2-4 alkenyl”. The alkenyl may be substituted or unsubstituted. In some embodiments, the alkenyl is C2, C3, C4, C5, C6, C7, or C8 alkenyl.
[0445] “Alkynyl” refers to an alkyl group as defined above that consists of at least two carbon atoms and at least one carbon-carbon triple bond; “C2-8 alkynyl” refers to a linear or branched alkynyl group containing 28 carbon atoms, including but not limited to ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, preferably “C2-6 alkynyl”, and more preferably “C2-4 alkynyl”. The alkynyl may be substituted or unsubstituted. In some embodiments, the alkynyl is C2, C3, C4, C5, C6, C7, or C8 alkynyl.
[0446] “-ylene” refers to a divalent group. For example, alkylene refers to a divalent alkyl group; alkenylene refers to a divalent alkenyl group; alkynylene refers to a divalent alkynyl group; cycloalkylene refers to a divalent cycloalkyl group; heterocycloalkylene refers to a divalent heterocycloalkyl group; arylene refers to a divalent aryl group; heteroarylene refers to a divalent heteroaryl group; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are as defined above; the “-ylene” groups may be optionally substituted or unsubstituted.
[0447] “Haloalkyl” refers to an alkyl group optionally substituted with one or more fluorine, chlorine, bromine, or iodine atoms, wherein the alkyl group is as defined above; non-limiting examples include difluoromethyl, dichloromethyl, dibromomethyl, trifluoromethyl, trichloromethyl, tribromomethyl, etc.
[0448] “Alkoxy” refers to -O-alkyl, wherein the alkyl is as defined above; non-limiting examples include methoxy, ethoxy, isopropoxy, tert-butoxy, etc.
[0449] In the present disclosure, “1 or more” refers to 1 or more than 1 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) under reasonable conditions.
[0450] Unless otherwise specified, as used in the present disclosure, the linking site of a substituent may be at any suitable position of the group to be substituted.
[0451] Also included within the scope of the present disclosure are metabolites of the compound of the present disclosure, i.e., substances formed in vivo upon administration of the compound of the present disclosure. Such products may result from, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymolysis, etc. of the administered compound. Thus, metabolites of the compound of the present disclosure include compounds prepared by a method of contacting the compound of the present disclosure with a mammal for a period of time sufficient to produce metabolic products thereof.
[0452] The present disclosure further includes within its scope prodrugs of the compound of the present disclosure, which are certain derivatives of the compound of the present disclosure that may themselves possess little or no toxicological or pharmacological activity and, when administered into or onto the body, may be converted (e.g., by hydrolytic cleavage) into the compound of the present disclosure having the desired activity. Typically, such prodrugs are functional derivatives of the compound that are readily convertible in vivo into the desired therapeutically active compound.
[0453] “Heteroatom or heteroatom group” refers to N, O, S, S(=O)2, S(O), etc.
[0454] “Cyano” refers to -CN.
[0455] “Hydroxy” refers to -OH.
[0456] “Sulfonyl” refers to -S(O)2-.
[0457] “Carboxyl” or “carboxylic acid” refers to -COOH.
[0458] “Oxo” refers to the =O group.
[0459] “Halogen” refers to fluorine, chlorine, bromine, or iodine.
[0460] “EA” or “EtOAc” refers to ethyl acetate.
[0461] “THF” refers to tetrahydrofuran.
[0462] “DCM” refers to dichloromethane.
[0463] “CataCXium A Pd-G3” refers to mesylate[(di(1-adamantyl)-n-butylphosphine)-2-(2'-amino-1,1'-biphenyl)]palladium(II).
[0464] “POCl3” refers to phosphorus oxychloride.
[0465] “NaHCO3” refers to sodium bicarbonate.
[0466] “NaCl” refers to sodium chloride.
[0467] “Na2SO4” refers to sodium sulfate.
[0468] “NH4Cl” refers to ammonium chloride.
[0469] “Cs2CO3” refers to cesium carbonate.
[0470] “CD3I” refers to deuterated iodomethane.
[0471] “NaBH(OAc)3” refers to sodium triacetoxyborohydride.
[0472] “Triton B” refers to benzyltrimethylammonium hydroxide.
[0473] “NaBH3CN” refers to sodium cyanoborohydride.
[0474] “CsF” refers to cesium fluoride.
[0475] “Na2CO3” refers to sodium carbonate.
[0476] “HOAc” refers to acetic acid.
[0477] “TPAP” refers to tetrapropylammonium perruthenate.
[0478] “NaH” refers to sodium hydride.
[0479] “i-PrOH” refers to isopropanol.
[0480] “DMF” refers to N,N-dimethylformamide.
[0481] “MeOH” refers to methanol.
[0482] “Pd / C” refers to palladium on carbon.
[0483] “DMSO” refers to dimethyl sulfoxide.
[0484] “TFA” refers to trifluoroacetic acid.
[0485] “DIEA” refers to N,N-diisopropylethylamine.
[0486] “Dess-Martin” refers to Dess-Martin reagent.
[0487] “PBS” refers to phosphate-buffered saline.
[0488] “SDS-PAGE” refers to sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
[0489] “PVDF” refers to polyvinylidene fluoride.
[0490] “PE” refers to petroleum ether.
[0491] “AcOH” refers to acetic acid.
[0492] “HCl” refers to hydrochloric acid.
[0493] “Dioxane” refers to 1,4-dioxane.
[0494] “ACN” refers to acetonitrile.
[0495] “DMP” refers to dimethyl phthalate.
[0496] “BH3” refers to borane.
[0497] “PdCl2(dtbpf)” refers to [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II).
[0498] “HATU” refers to O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate.
[0499] “EtOH” refers to ethanol.
[0500] “DMAP” refers to 4-dimethylaminopyridine.
[0501] “TBAF” refers to tetrabutylammonium fluoride.
[0502] “TMEDA” refers to tetramethylethylenediamine.
[0503] “s-BuLi” refers to tert-butyllithium.
[0504] “NaBH4” refers to sodium borohydride.
[0505] “CH3I” refers to iodomethane.
[0506] “DEA” refers to diethanolamine.
[0507] “EDCI” refers to 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride.
[0508] “HOBT” refers to 1-hydroxybenzotriazole.
[0509] “LAH” refers to lithium aluminum hydride.
[0510] “TsCl” refers to p-toluenesulfonyl chloride.
[0511] “Na2S2O3” refers to sodium thiosulfate.
[0512] “NaNO2” refers to sodium nitrite.
[0513] “BBr3” refers to boron tribromide.
[0514] “K3PO4” refers to potassium phosphate.
[0515] “AcOK” refers to potassium acetate.
[0516] “DIBAL-H” refers to diisobutylaluminum hydride.
[0517] “TEA” refers to triethylamine.
[0518] “GTP” refers to guanosine triphosphate.
[0519] “Prep-HPLC” or “pre-HPLC” refers to preparative high-performance liquid chromatography.
[0520] “NFSI” refers to N-fluorobenzenesulfonimide.
[0521] “Ti(O-iPr)4” refers to titanium(IV) isopropoxide.
[0522] “Pd(dppf)Cl2” refers to [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride.
[0523] “t-BuOK” refers to potassium tert-butoxide.
[0524] “Grubbs II catalyst” refers to ferrocenedimethylruthenium(II) bis(diisopropylphosphine).
[0525] “Pd(OH)2” refers to palladium hydroxide.
[0526] “LiAlD4” refers to lithium aluminum deuteride.
[0527] “CH3MgBr” refers to methylmagnesium bromide.
[0528] “BOPCl” refers to bis(2-oxo-3-oxazolidinyl)phosphinic chloride.
[0529] “m-CPBA” refers to m-chloroperoxybenzoic acid.
[0530] “Ti(Oi-Pr)4” refers to tetraisopropyl titanate.
[0531] “T3P” refers to propylphosphonic anhydride.
[0532] “sat.” refers to a saturated solution.
[0533] “aq” refers to an aqueous solution.
[0534] In the chemical structures herein, “-” as a linking bond represents a single bond, and “=” as a linking bond represents a double bond (in the case of an undefined configuration, it may be trans or cis).
[0535] In the present disclosure, the expression “-N(group)1-2” refers to -NH(group) or -N(group)2.
[0536] “Optional” or “optionally” means that the subsequently described event or circumstance may, but does not necessarily, occur; this description includes instances where the event or circumstance occurs and instances where it does not. For example, “a heterocycloalkyl group optionally substituted with an alkyl group” means that the alkyl group may, but does not necessarily, exist; this description includes instances where the heterocycloalkyl group is substituted with the alkyl group and instances where it is not.
[0537] “Substituted” means that one or more, preferably up to 5, and more preferably 1-3, hydrogen atoms in the group are each independently replaced by a corresponding number of substituents. It goes without saying that substituents are only positioned at their chemically feasible positions, and those skilled in the art can determine (experimentally or theoretically) feasible or infeasible substitutions without undue effort. For example, it may be unstable when an amino or hydroxy group with a free hydrogen is bound to a carbon atom with an unsaturated (e.g., olefinic) bond.
[0538] Unless otherwise indicated, the term “optionally substituted” as used herein means that a group may be unsubstituted or substituted; when it is substituted, the substituent may be one or more (e.g., 2, 3, 4, 5, or 6) groups independently selected from alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, haloalkyl, alkoxy, amino, aminoalkyl, cyano, halogen, oxo, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein the alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, haloalkyl, alkoxy, amino, and aminoalkyl are optionally substituted with one or more (e.g., 2, 3, 4, 5, or 6) cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one or more groups selected from alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, haloalkyl, alkoxy, amino, aminoalkyl, cyano, halogen, and oxo.
[0539] Unless otherwise indicated, the following terms and phrases as used herein are intended to have the following meanings. A specific term or phrase, in the absence of a particular definition, should not be considered indefinite or unclear, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to the corresponding commercial product or its active ingredient.
[0540] “Pharmaceutical composition” refers to a mixture containing one or more of the compounds or the physiologically / pharmaceutically acceptable salts or prodrugs thereof described herein and other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The pharmaceutical composition is intended to promote the administration to an organism, so as to facilitate the absorption of the active ingredient, thereby exerting biological activity.
[0541] The present disclosure further provides a pharmaceutically acceptable salt of the compound of formula (I). The term “pharmaceutically acceptable salt” refers to a relatively non-toxic acid addition salt or base addition salt of the compound of the present disclosure. The acid addition salt is a salt formed by the compound of formula (I) of the present disclosure with a suitable inorganic acid or organic acid. Such salts may be prepared during the final isolation and purification of the compound, or by reacting the purified compound of formula (I) in its free base form with a suitable organic acid or inorganic acid. Representative acid addition salts include hydrochloride, tartrate, hydrobromide, sulfate, bisulfate, sulfite, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, biphosphate, carbonate, bicarbonate, toluate, citrate, maleate, fumarate, succinate, benzoate, methanesulfonate, p-toluenesulfonate, gluconate, lactobionate, lauryl sulfonate, etc. The base addition salt is a salt formed by the compound of formula (I) with a suitable inorganic base or organic base, including, for example, salts formed with alkali metal, alkaline earth metal, or quaternary ammonium cations, such as sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, tetramethyl quaternary ammonium salts, tetraethyl quaternary ammonium salts, etc.; amine salts include salts formed with ammonia (NH3), primary amines, secondary amines, or tertiary amines, such as methylamine salts, dimethylamine salts, trimethylamine salts, triethylamine salts, ethylamine salts, etc.
[0542] The pharmaceutically acceptable salts of the present disclosure can be synthesized from a parent compound containing an acidic or basic group using conventional chemical methods. In general, such salts are prepared by subjecting the compounds in their free acid or base form to a reaction with a stoichiometric amount of an appropriate base or acid in water, an organic solvent, or a mixture thereof.
[0543] The term “pharmaceutically acceptable excipient” refers to an inert substance administered together with an active ingredient to facilitate administration of the active ingredient, including but not limited to any acceptable glidants, sweeteners, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants, wetting agents, dispersants, disintegrants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers approved by the National Medical Products Administration, PRC, for use in humans or animals (e.g., livestock).
[0544] As used herein and as is familiar in the art, “treatment” or “treating” is a method for obtaining beneficial or desired results, including clinical results. The beneficial or desired clinical results may include, but are not limited to, reducing tumor progression, decreasing tumor size, lowering tumor growth rate, reducing tumor invasive and metastatic potential, alleviating or ameliorating one or more symptoms or conditions, diminishing the extent of disease, stabilizing (i.e., not worsening) the disease state, preventing disease spread, delaying or slowing down disease progression, ameliorating or mitigating the disease state, and achieving remission (whether partial or complete), whether detectable or undetectable. “Treatment” or “treating” may also refer to prolonging survival as compared to the expected survival if not receiving treatment.
[0545] The therapeutic dose of the compound of the present application may be determined based on, for example, the specific use of the treatment, the mode of administration of the compound, the health and state of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compound of the present application in a pharmaceutical composition may not be fixed and depends on various factors including dose, chemical properties (e.g., hydrophobicity), and route of administration.
[0546] The term “treatment” or “treating” refers to administering the compound or formulation described in the present application to ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:
[0547] (i) inhibiting the disease or disease state, i.e., arresting its progression; and
[0548] (ii) alleviating the disease or disease state, i.e., causing regression of the disease or disease state.
[0549] The term “therapeutically effective amount” refers to an amount of the compound of the present application for (i) treating a specific disease, condition, or disorder, (ii) alleviating, ameliorating, or eliminating one or more symptoms of a specific disease, condition, or disorder, or (iii) preventing or delaying onset of one or more symptoms of the specific disease, condition, or disorder described herein. The amount of the compound of the present application constituting the “therapeutically effective amount” varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but may be determined routinely by those skilled in the art in accordance with their knowledge and the present disclosure.
[0550] Unless otherwise required in the present application, throughout the specification and the appended claims, the word “comprise” and variations thereof such as “comprises” and “comprising” should be interpreted in an open and inclusive sense, meaning “including, but not limited to”.
[0551] Throughout this specification, references to “in some embodiments”, “in an embodiment”, “in another embodiment”, or “in certain embodiments” mean that a specific referenced element, structure, or feature described in connection with the embodiment is included in at least one embodiment. Thus, the phrase “in some embodiments”, “in an embodiment”, “in another embodiment”, or “in certain embodiments” appearing at various places throughout the specification do not necessarily all refer to the same embodiment. Furthermore, the specific elements, structures, or features may be combined in any suitable manner in one or more embodiments.
[0552] Unless otherwise indicated, the term “isomer” is intended to include geometric isomers, cis-trans isomers, stereoisomers, enantiomers, optical isomers, diastereomers, and tautomers.
[0553] The compounds of the present disclosure may be in the form of a specific geometric isomer or stereoisomer. All such compounds are contemplated herein, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically enriched or diastereomerically enriched mixtures, all of which fall within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All such isomers and mixtures thereof are encompassed within the scope of the present disclosure.
[0554] Unless otherwise indicated, the term “enantiomer” or “optical isomer” refers to stereoisomers that are mirror images of each other.
[0555] Unless otherwise indicated, the term “cis-trans isomer” or “geometric isomer” arises due to the inability of free rotation caused by a double bond or a single bond of a ring-forming carbon atom.
[0556] Unless otherwise indicated, the term “diastereomer” refers to stereoisomers whose molecules have two or more chiral centers and are not mirror images of each other.
[0557] Unless otherwise indicated, “(+)” denotes dextrorotation, “(-)” denotes levorotation, and “(±)” denotes racemization.
[0558] The term “prodrug” refers to a chemical derivative of the compound of the present disclosure, which, upon undergoing a chemical reaction in vivo, is converted into the compound of general formula I.
[0559] “Optional” or “optionally” means that the subsequently described event or circumstance may, but does not necessarily, occur, and the description includes instances where the event or circumstance occurs and instances where it does not.
[0560] The term “substituted” means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which may include deuterium and hydrogen variants, provided that the valence of the specific atom is normal and the resulting substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Substitution with oxygen does not occur on aromatic groups.
[0561] The term “optionally substituted” means that a group may or may not be substituted with a substituent. Unless otherwise specified, the type and number of the substituent may be arbitrary as long as being chemically achievable.
[0562] When any variable (e.g., R) occurs once or more in the constitution or structure of a compound, the definition of the variable in each case is independent. Thus, for example, if a group is substituted with 0-2 R, the group may be optionally substituted with up to two R, and the definition of R in each case is independent. Furthermore, combinations of substituents and / or variants thereof are permissible only if such combinations result in a stable compound.
[0563] When the number of a linking group is 0, for example, -(CRR)0-, it means that the linking group is a single bond.
[0564] When one of the variables is selected from a single bond, it means that the two groups connected by it are linked directly. For example, in A-L-Z, if L represents a single bond, it means that the structure is actually A-Z.
[0565] When a substituent is absent, it means that the substituent does not exist. For example, in A-X, if X is absent, it means that the structure is actually A. When the atom through which a listed substituent is linked to the group to be substituted is not specified, the substituent may be linked via any atom of the group. For example, pyridyl as a substituent may be linked to the group to be substituted via any carbon atom on the pyridine ring.
[0566] When the linking direction of a listed linking group is not specified, its linking direction is arbitrary. For example, if the linking group L is -M-W-, then -M-W- may either link ring A to ring B in a direction same as the left-to-right reading order to form the final structure, or link ring A to ring B in a direction opposite to the left-to-right reading order to form the final structure. Combinations of such linking groups, substituents, and / or variants thereof are permissible only if they result in a stable compound.
[0567] Unless otherwise indicated, when a group has one or more linkable sites, any one or more of the sites of the group may be linked to other groups via chemical bonds. If the linking mode for chemical bonds is non-specific and H atoms are present at a linkable site, when the linkable site is linked to chemical bonds, the number of the H atoms at the linkable site will be correspondingly reduced based on the number of the linked chemical bonds, resulting in a group with the corresponding valence. The chemical bond that links the site to another group may be represented by a straight solid bond \ a straight dashed bond \ or a wavy line ) .For example, the straight solid bond in -OCH2CH3 refers to linkage to another group "S. • • • • H C CH3 via the oxygen atom in the group; the straight dashed bonds in 3 refer to linkage to other groups via two ends of the sulfur atom in the group; the wavy lines in refer to linkage to other groups via NH . . . . . the carbon atoms at positions 1 and 2 in the phenyl group; 1 means that any linkable site on the pyrrolidinyl group may be linked to another group via 1 chemical bond, encompassing at least 3 linking modes: groups in the NH , and ; even if an H atom is depicted on -N-, ' still encompasses linking mode; however, if the linkable site is linked to 1 chemical bond, the number of H at that site will be correspondingly reduced by 1, resulting in the corresponding monovalent pyrrolidinyl group.
[0568] Unless otherwise indicated, the wedged solid bond and the wedged dashed bond are used to denote the absolute configuration of a stereogenic center, and the straight solid bond and the straight dashed bond '' are used to denote the relative configuration of a stereogenic center; the wavy line is used to represent either a wedged solid bond or a wedged dashed bond or a straight , or the wavy line is used to represent either a straight solid bond or or ,Bn the absolute configuration of a stereogenic center is denoted by a
[0569] Unless otherwise indicated, wedged solid bond and a wedged dashed bond, and the relative configuration of a stereogenic center is denoted by a straight solid bond and a straight dashed bond; a wavy line represents either a wedged solid bond or a wedged dashed bond, or a wavy line represents either a straight solid bond or a straight dashed bond.
[0570] Unless otherwise indicated, when a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, is present in a compound, and each atom on the double bond is linked to two different substituents (for a double bond involving a nitrogen atom, a lone pair of electrons on the nitrogen atom is regarded as one substituent linked thereto), if an atom on the double bond in the compound is linked to its substituent using a wavy line ( ? \ it means that the compound includes a (Z)-isomer and an (E )-isomer.For example, the following formula (A) indicates that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2), or in the form of a mixture of both isomers of formula (A-1) and formula (A-2); the following formula (B) indicates that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2), or in the form of a mixture of both isomers of formula (B-1) and formula (B-2). The following formula (C) indicates that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2), or in the form of a mixture of both isomers of formula (C-1) and formula (C-2). differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structures of the present disclosure, except for those in which hydrogen is replaced by deuterium or tritium, or carbon is replaced by 13C or 14C enriched carbon, fall within the scope of the present disclosure.
[0572] Unless otherwise indicated, in some cases, the compounds described in the present disclosure exist as diastereomers, enantiomers, or other stereoisomers. When the absolute stereochemistry is not specified, the compounds provided by the present disclosure include all diastereomers, enantiomers, and epimeric forms, as well as suitable mixtures thereof. Isolation of stereoisomers can be carried out by chromatography, or by formation of diastereomers followed by separation via recrystallization or chromatography or any combination thereof. Stereoisomers can also be obtained by stereoselective synthesis.
[0573] Unless otherwise indicated, the term “atropisomer (or atropoisomer)” refers to a stereoisomer with a specific spatial configuration resulting from restricted rotation about a single bond due to high steric hindrance. Certain compounds of the present disclosure may exist as atropisomers. The compounds disclosed herein include all atropisomers, which may be pure individual atropisomers, or enriched in one of the atropisomers, or the respective non-specific mixtures. If the rotational barrier about a single bond is sufficiently high and the interconversion between conformers is sufficiently slow, isolation of the isomers can be allowed. For example, OH OH and OH are a pair of atropisomers, wherein the ' on the naphthalene ring indicates that the stereochemical orientation of that side is outward, and the ■' indicates that the stereochemical orientation of that side is inward. Unless otherwise indicated, some embodiments of the present disclosure, for example,
[0574] in ocd3 represents or represents N OH OH OH represents H N or OH H N .CD3 represents represents OH . H N or OH
[0575] Unless otherwise indicated, in some embodiments of the present disclosure, and are interchangeable, wherein Ra and Rc are as defined and described in the present OCD3 and
[0576] Unless otherwise specified, disclosure; for example, are interchangeable and represent OH is used to indicate that a hydrogen atom at any position of a group within may be replaced.
[0577] Unless otherwise indicated, the term “enriched in one isomer”, “isomerically enriched”, “enriched in one enantiomer”, or “enantiomerically enriched” means that the content of one of the isomers or enantiomers is less than 100%, and the content of the isomer or enantiomer is more than or equal to 60%, or more than or equal to 70%, or more than or equal to 80%, or more than or equal to 90%, or more than or equal to 95%, or more than or equal to 96%, or more than or equal to 97%, or more than or equal to 98%, or more than or equal to 99%, or more than or equal to 99.5%, or more than or equal to 99.6%, or more than or equal to 99.7%, or more than or equal to 99.8%, or more than or equal to 99.9%.
[0578] Unless otherwise indicated, the term “isomeric excess” or “enantiomeric excess” refers to the difference in relative percentages between two isomers or two enantiomers. For example, if the content of one of the isomers or enantiomers is 90% and the content of the other isomer or enantiomer is 10%, the isomeric or enantiomeric excess (ee value) is 80%.
[0579] Optically active (R)- and (S)-enantiomers, as well as D- and L-isomers, may be prepared by chiral synthesis, chiral reagents, or other conventional techniques. If one enantiomer of a certain compound of the present disclosure is desired, it may be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to afford the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), salts of diastereomers are formed with an appropriate optically active acid or base, and then resolution of the diastereomers is performed by conventional methods known in the art, followed by recovery of the pure enantiomers. In addition, separation of enantiomers and diastereomers is generally accomplished by chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amines).
[0580] The compound of the present disclosure may contain a non-natural proportion of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compound may be labeled with a radioisotope, such as tritium (3H), iodine-125 (125I), or C-14 (14C). For another example, hydrogen may be replaced by deuterium to form a deuterated drug, and the bond formed between deuterium and carbon is firmer than that formed between ordinary hydrogen and carbon. Compared to a non-deuterated drug, the deuterated drug offers advantages such as reduced toxic and side effects, increased pharmaceutical stability, enhanced efficacy, and prolonged biological half-life. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure.
[0581] Unless otherwise indicated, D described in the present disclosure refers to deuterium (2H).
[0582] Unless otherwise indicated, the term “leaving group” refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chlorine, bromine, iodine; sulfonate groups, such as methanesulfonate, benzenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate, etc.; and acyloxy groups, such as acetoxy, trifluoroacetoxy, etc.
[0583] Unless otherwise indicated, the term “protecting group” includes, but is not limited to, an “amino protecting group”, a “hydroxy protecting group”, or a “sulfhydryl protecting group”. The term “amino protecting group” refers to a protecting group suitable for use in preventing side reactions at the nitrogen position of an amino group. Representative amino protecting groups include, but are not limited to: formyl; acyl groups, such as alkanoyl groups (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl groups, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl groups, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl groups, such as benzyl (Bn), trityl (Tr), and 1,1-di-(4'-methoxyphenyl)methyl; and silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term “hydroxy protecting group” refers to a protecting group suitable for use in preventing side reactions of a hydroxy group. Representative hydroxy protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanoyl groups (e.g., acetyl); arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (benzhydryl, DPM); and silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc.
[0584] The compounds of the present disclosure may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent alternatives well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of the present disclosure.
[0585] The structures of the compounds of the present disclosure may be confirmed by conventional methods well known to those skilled in the art. If the present disclosure relates to an absolute configuration of a compound, the absolute configuration may be confirmed by means of conventional techniques in the art. For example, in single crystal X-ray diffraction (SXRD), diffraction intensity data of the single crystal grown are collected with a Bruker D8 venture diffractometer, with the light source being Cu-Ka radiation and the scanning mode being scanning; after the relevant data are collected, a direct method (Shelxs97) is further employed to analyze the crystal structure, and thus the absolute configuration can be confirmed.
[0586] The compound or the pharmaceutically acceptable salt thereof of the present disclosure may be administered to mammals including humans via oral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), topical (powder, ointment, or drop), or intratumoral administration.
[0587] The compound of the present disclosure may be administered at a dose of about 0.05-300 mg / kg body weight / day, preferably 10-300 mg / kg body weight / day, and more preferably 10-200 mg / kg body weight / day.
[0588] The compound or the pharmaceutically acceptable salt thereof of the present disclosure may be formulated into a solid dosage form for oral administration, including but not limited to capsules, tablets, pills, pulvis, granules, etc. In these solid dosage forms, the compound of formula (I) of the present disclosure, as an active ingredient, is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (1) fillers or bulking agents, such as starch, lactose, sucrose, glucose, mannitol, silicic acid, etc.; (2) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, acacia, etc.; (3) humectants, such as glycerol, etc.; (4) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, sodium carbonate, etc.; (5) solution retarders, such as paraffin, etc.; (6) absorption accelerators, such as quaternary ammonium compounds, etc.; (7) wetting agents, such as cetyl alcohol, glycerol monostearate, etc.; (8) adsorbents, such as kaolin, etc.; and (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc., or mixtures thereof. Capsules, tablets, and pills may further comprise buffers.
[0589] The solid dosage forms such as tablets, dragees, capsules, pills, and granules may be coated or microencapsulated with coating and shell materials such as enteric coatings and other materials well known in the art. They may comprise opacifying agents, and the active ingredient in such a composition may be released in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be used are polymeric substances and wax-based substances. If necessary, the active ingredient can also be in microcapsule form with one or more of the above-mentioned excipients.
[0590] The compound or the pharmaceutically acceptable salt thereof of the present disclosure may be formulated into a liquid dosage form for oral administration, including but not limited to pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, etc. In addition to the compound of formula (I) or the pharmaceutically acceptable salt thereof as an active ingredient, the liquid dosage form may comprise inert diluents conventionally used in the art, such as water and other solvents, solubilizers, and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, etc., or mixtures of these substances, etc. In addition to these inert diluents, the liquid dosage form of the present disclosure may further comprise conventional adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, perfuming agents, etc.
[0591] The suspending agents include, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminium methanolate, agar, etc., or mixtures of these substances.
[0592] The compound or the pharmaceutically acceptable salt thereof of the present disclosure may be formulated into a dosage form for parenteral injection, including but not limited to physiologically acceptable sterile aqueous or water-free solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.
[0593] The compound or the pharmaceutically acceptable salt thereof of the present disclosure may also be formulated into a dosage form for topical administration, including, for example, ointments, pulvis, suppositories, drops, sprays, inhalants, etc. The compound of formula (I) or the pharmaceutically acceptable salt thereof of the present disclosure as an active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and optionally a preservative, buffer, or, if necessary, a propellant that may be required.
[0594] The present disclosure further provides a pharmaceutical composition, comprising the compound of formula (I) or the pharmaceutically acceptable salt thereof of the present disclosure as an active ingredient, and a pharmaceutically acceptable carrier, excipient, or diluent. In the preparation of the pharmaceutical composition, the compound of formula (I) or the pharmaceutically acceptable salt thereof of the present disclosure is typically mixed with the pharmaceutically acceptable carrier, excipient, or diluent.
[0595] The composition of the present disclosure may be formulated into a conventional pharmaceutical formulation according to a conventional preparation method. Examples include tablets, pills, capsules, pulvis, granules, emulsions, suspensions, dispersions, solutions, syrups, elixirs, ointments, drops, suppositories, inhalants, sprays, etc.
[0596] The compound or the pharmaceutically acceptable salt thereof described herein may be administered alone or, if desired, in combination with an additional pharmaceutically acceptable therapeutic agent, such as an additional anti-tumor drug. The ingredients to be combined may be administered simultaneously or sequentially, in a single formulation form or in different formulation forms. The combination may include not only a combination of the compound of the present disclosure and one additional active agent, but also a combination of the compound of the present disclosure and two or more additional active agents.
[0597] In the present disclosure, the additional pharmaceutically acceptable therapeutic agent that may be used together with or in combination with the KRAS G12D degrader, the compound of formula (I), may be: EGFR and / or a mutant inhibitor thereof, ErbB2 (Her2) and / or a mutant inhibitor thereof, ALK and / or a mutant inhibitor thereof, MEK and / or a mutant inhibitor thereof, KRAS and / or a mutant inhibitor thereof, BCR-ABL and / or a mutant inhibitor thereof, FGFR1 / FGFR2 / FGFR3 and / or a mutant inhibitor thereof, ROS1 and / or a mutant inhibitor thereof, c-MET and / or a mutant inhibitor thereof, AXL and / or a mutant inhibitor thereof, NTRK1 and / or a mutant inhibitor thereof, RET and / or a mutant inhibitor thereof, taxane, a platinum-containing compound, an antimetabolite, a mitotic kinase inhibitor, an immunotherapeutic agent, an anti-angiogenic drug, a topoisomerase inhibitor, A-Raf / B-Raf / C-RAf and / or a mutant inhibitor thereof, ERK and / or a mutant inhibitor thereof, an apoptosis inhibitor, AKT and / or a mutant inhibitor thereof, an mTOR inhibitor, an epigenetic regulator, an IGF1 / 2 and / or IGF1-R inhibitor, Ras GEF and / or a mutant inhibitor thereof, SOS1 and / or a mutant inhibitor thereof, SHP2 and / or a mutant inhibitor thereof, PI3K and / or a mutant inhibitor thereof, or a PD-1 / PD-L1 inhibitor; EGFR and / or a mutant degrader thereof, ErbB2 (Her2) and / or a mutant degrader thereof, ALK and / or a mutant degrader thereof, MEK and / or a mutant degrader thereof, KRAS and / or a mutant degrader thereof, BCR-ABL and / or a mutant degrader thereof, FGFR1 / FGFR2 / FGFR3 and / or a mutant degrader thereof, ROS1 and / or a mutant degrader thereof, c-MET and / or a mutant degrader thereof, AXL and / or a mutant degrader thereof, NTRK1 and / or a mutant degrader thereof, RET and / or a mutant degrader thereof, A-Raf / B-Raf / C-RAf and / or a mutant degrader thereof, ERK and / or a mutant degrader thereof, AKT and / or a mutant degrader thereof, an IGF1 / 2 and / or IGF1-R degrader, Ras GEF and / or a mutant degrader thereof, SOS1 and / or a mutant degrader thereof, SHP2 and / or a mutant degrader thereof, or PI3K and / or a mutant degrader thereof; EGFR and / or a mutant monoclonal antibody thereof, ErbB2 (Her2) and / or a mutant monoclonal antibody thereof, a PD-1 / PD-L1 monoclonal antibody, or a CTLA-4 monoclonal antibody; a PD-L1 / TIGHT bispecific antibody, a PD-L1 / CTLA-4 bispecific antibody, an EGFR / MET bispecific antibody, an EGFR / CD3 bispecific antibody, an EGFR / 4-IBB bispecific antibody, a PD-L1 / 4-IBB bispecific antibody, or an HER2 / CD3 bispecific antibody.
[0598] In the present disclosure, the additional pharmaceutically acceptable therapeutic agent that may be used together with or in combination with the KRAS G12D degrader, the compound of formula (I), may be: afatinib, erlotinib, gefitinib, lapatinib, cetuximab, panitumumab, osimertinib, olmutinib, EGF-816, trastuzumab, pertuzumab, crizotinib, alectinib, entrectinib, brigatinib, trametinib, cobimetinib, binimetinib, selumetinib, refametinib, imatinib, dasatinib, nilotinib, nintedanib, crizotinib, lorlatinib, ceritinib, merestinib, paclitaxel, nab-paclitaxel, docetaxel, cisplatin, carboplatin, oxaliplatin, 5-fluorouracil, capecitabine, floxuridine, cytarabine, gemcitabine, a combination of trifluridine and tipiracil (=TAS102), palbociclib, ribociclib, abemaciclib, ipilimumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab, pidilizumab, PDR-001 (=spartalizumab), bevacizumab, irinotecan, liposomal irinotecan, topotecan, ulixertinib, rapamycin, temsirolimus, everolimus, ridaforolimus, JQ-1, GSK 525762, OTX 015 (=MK8628), CPI 0610, TEN-010 (=RO6870810), xentuzumab (antibody 60833 in WO 2010 / 066868), or MEDI-573 (=dusigitumab).
[0599] On the basis of conforming to common general knowledge in the art, the preferred conditions described above may be combined arbitrarily to obtain preferred embodiments of the present disclosure.
[0600] The reagents and starting materials used in the present disclosure are all commercially available.
[0601] Compounds are named according to conventional nomenclature rules in the art, and commercially available compounds may also be referred to by the catalog names provided by suppliers.
[0602] According to the present disclosure, a KRAS G12D kinase activity assay demonstrates that the compound of formula I described in the present disclosure is capable of effectively binding to the KRAS G12D target protein or producing an inhibitory effect, and a Western-Blot analysis demonstrates that the compound of formula I described in the present disclosure is capable of effectively and specifically degrading the KRAS G12D protein in A427 cells. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the deuteride, the hydrate, the solvate, the metabolite, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof described in the present disclosure can effectively degrade the KRAS G12D protein, thereby achieving the effect of preventing or treating a disease or disorder associated with KRAS G12D or caused by the interaction between KRAS G12D and the SOS1 or SHP2 protein. DETAILED DESCRIPTION
[0603] The following examples further illustrate the present disclosure, but the present disclosure is not limited thereto. Experimental procedures without specific conditions indicated in the following examples were conducted in accordance with conventional procedures and conditions, or in accordance with product instructions.
[0604] The present disclosure is further explained in detail and comprehensively below with reference to examples; however, the examples are not intended to limit the present disclosure, and the present disclosure is not limited to the contents of the examples. The starting materials in the examples of the present disclosure are known and commercially available, or may be synthesized by using or according to methods known in the art. Unless otherwise stated, experimental procedures without specific conditions indicated in the examples of the present disclosure were generally conducted under conventional conditions or conditions recommended by the manufacturers of the starting materials or commercial products. I. Compound Preparation Examples
[0605] Intermediate 1: tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(piperazin-1-ylmethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0606] Step 1: preparation of 1-((4-benzylpiperazin-1-yl)methyl)cyclopropyl)methanol
[0607] 1,1-Cyclopropanedicarboxylic acid monomethyl ester (14.7 g, 102.0 mmol) was dissolved in toluene (120 mL), and the mixture was heated to 120 °C. Then, 1-tert-butoxycarbonyl-piperazine (12.6 g, 68.0 mmol) and phenylsilane (5.5 g, 51.0 mmol) were added. The mixture was reacted at 120 °C for 16 h, and then phenylsilane (14.7 g, 136.0 mmol) and zinc acetate (1.2 g, 6.8 mmol) were added. The resulting mixture was reacted for another 2 h. After the reaction was completed, the reaction mixture was cooled to room temperature. A 2 M aqueous hydrochloric acid solution (120 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL x 3). The aqueous phase was adjusted to pH 12 with a 4 M sodium hydroxide solution (60 mL) and extracted with dichloromethane (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography to give 1-((4-benzylpiperazin-1-yl)methyl)cyclopropyl)methanol.
[0608] LC-MS: (ESI, m / z): [M+H]+ = 261.2.
[0609] Step 2: preparation of tert-butyl 3-(2-((1-((4-benzylpiperazin-1-yl)methyl)cyclopropyl)methoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0610] tert-Butyl 3-(2,7-dichloro-8-fluoropyrido[4,3- ^]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate (5.00 g, 11.70 mmol) and 1-((4-benzylpiperazin-1-yl)methyl)cyclopropyl)methanol (3.30 g, 12.80 mmol) were dissolved in THF, and then cesium carbonate was added. The mixture was stirred at 90 °C for 16 h. After the reaction was completed, the reaction mixture was cooled to room temperature. A saturated ammonium chloride solution (100 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 3:1) to give tert-butyl 3-(2-((1-((4-benzylpiperazin-1-yl)methyl)cyclopropyl)methoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0611] LC-MS: (ESI, m / z): [M+H]+ = 652.3.
[0612] Step 3: preparation of tert-butyl 3-(7-chloro-8-fluoro-2-((1-(piperazin-1- ylmethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.5 g, 2.4 mmol) was dissolved in DCM (15 mL), and 1-chloroethyl chloroformate (1.03 g, 7.2 mmol) and DIEA (0.62 g, 4.8 mmol) were added. The mixture was reacted at room temperature for 30 min and concentrated under reduced pressure. The concentrate was dissolved in MeOH (15 mL), and the resulting mixture was heated to 50 °C and reacted for 30 min. After the reaction was completed, the mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 3-(7-chloro-8-fluoro-2-((1-(piperazin-1-ylmethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0614]
[0615] LC-MS: (ESI, m / z): [M / 2+H]+ = 562.2. Step 4: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(piperazin-1-ylmethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate TIPS _ OMOM
[0616] cataCXium A Pd-G3 (262 mg, 0.36 mmol) and cesium carbonate (1.74 g, 5.34 mmol) were added to a mixed solution of tert-butyl 3-(7-chloro-8-fluoro-2-((1-(piperazin-1- ylmethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.00 g, 1.78 mmol) and ((2-fluoro-6-((methoxymethyl)oxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (1.1 g, 2.14 mmol) in dioxane (9 mL) and H2O (1 mL). The mixture was reacted at 80 °C for 16 h under nitrogen atmosphere. Water (100 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by column chromatography (DCM:MeOH = 100:1 to 7:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(piperazin-1-ylmethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0617] LC-MS: (ESI, m / z): [M+H]+= 912.4.
[0618] 1H NMR (400 MHz, DMSO-d6) 5 9.17 (s, 1H), 8.11 (dd, J = 9.2 Hz, 6.0 Hz, 1H), 7.75 (d, J = 2.4 Hz, 1H), 7.59-7.53 (m, 1H), 7.33 (d, J = 2.4 Hz, 1H), 5.37 (s, 2H), 4.77-4.70 (d, J = 12 Hz, 1H), 4.40-4.15 (m, 6H), 3.73 (d, J = 11.6 Hz, 1H), 3.43 (s, 1H), 3.38-3.32 (m, 2H), 2.95-2.85 (m, 4H), 2.52-2.51 (m, 2H), 2.472.29 (m, 2H), 1.96-1.77 (m, 3H), 1.74-1.57 (m, 2H), 1.46 (s, 9H), 0.81 (t, J = 7.2 Hz, 18H), 0.69-0.60 (m, 2H), 0.53-0.37 (m, 5H).
[0619] Intermediate 2: intermediate 2-P1 and intermediate 2-P2
[0620] Step 1: preparation of tert-butyl 3-benzyl-1-formyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0621] tert-Butyl 3-benzyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (9.5 g, 31.4 mmol) and TMEDA (4.1 g, 35.16 mmol) were dissolved in tetrahydrofuran (100 mL), and the mixture was cooled to 0 °C. 5-BuLi (36 mL, 47.1 mmol, 1.3 mol / L solution in n-hexane) was added, and the mixture was reacted at 0 °C for 1 h. DMF (2.8 g, 37.7 mmol) was then added, and the mixture was slowly warmed to room temperature and reacted for 10 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (100 mL), and the mixture was extracted with ethyl acetate (100 mL x 2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated to give a crude product. The crude product was purified by silica gel column chromatography (PE:EtOAc = 100:1 to 20:1) to give tert-butyl 3-benzyl-1-formyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0622] LC-MS: (ESI, m / z): [M+H]+ = 331.2.
[0623] Step 2: preparation of tert-butyl 3-benzyl-1-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0624] tert-Butyl 3-benzyl-1-formyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (5.0 g, 15.15 mmol) was dissolved in methanol (500 mL), and NaBH4 (1.15 g, 30.30 mmol) was added at 0 °C. The mixture was reacted for 3 h. After the reaction was completed, a saturated aqueous ammonium chloride solution (20 mL) was added, and the mixture was extracted with ethyl acetate (60 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure to give tert butyl 3-benzyl-1-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0625] LC-MS: (ESI, m / z): [M+H]+ = 333.2.
[0626] Step 3: preparation of tert-butyl 3-benzyl-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate Boc N i Bn OH
[0627] tert-Butyl 3-benzyl-1-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (4.1 g, 12.30 mmol) was dissolved in N,N-dimethylformamide (50 mL), and the mixture was cooled to 0 °C. NaH (733 mg, 18.45 mmol) was added, and the mixture was reacted at 0 °C for 1 h. CH3I (5.24 g, 36.90 mmol) was then added, and the mixture was reacted at 25 °C for 3 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (40 mL), and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (PE:EA = 4:1) to give tert-butyl 3-benzyl-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0628] LC-MS: (ESI, m / z): [M+H]+ = 347.2.
[0629] Step 5: preparation of tert-butyl 1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0630] tert-Butyl 3-benzyl-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (3.1 g, 8.95 mmol) was dissolved in methanol (30 mL), and Pd / C (0.5 g) was added. The mixture was reacted at 25 °C for 16 h under hydrogen atmosphere. The reaction liquid was filtered through celite, and the filtrate was concentrated to give tert-butyl 1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0631] LC-MS: (ESI, m / z): [M-Boc+H] + = 157.3.
[0632] Step 6: preparation of intermediate 2-P1 and intermediate 2-P2 Boc
[0633] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (1.76 g, 7.00 mmol) and triethylamine (2.12 g, 21.00 mmol) were dissolved in tetrahydrofuran (30 mL), and the mixture was cooled to -78 °C. tert-Butyl 1- (methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.80 g, 7.00 mmol) was added, and the mixture was reacted at -78 °C for 3 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (50 mL), and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate. The compound obtained from the purification was subjected to SFC resolution with the following method: chromatographic column: Daicel chiralpak OD_3, 3 x 150 mm, 3 nm; mobile phase: CO2 / MeOH (0.1% DEA), gradient: 80 / 20, 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C.
[0634] Intermediate 2-P1 (chiral column retention time: 2.248 min). SFC analysis method: column: Daicel chiralpak AD_3, 3 x 150 mm, 3 ^m; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 50 / 50, 6 min; flow rate: 1.5 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0635] Intermediate 2-P2 (chiral column retention time: 2.350 min). SFC analysis method: column: Daicel chiralpak AD_3, 3 x 150 mm, 3 nm; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 50 / 50, 6 min; flow rate: 1.5 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0636] LC-MS: (ESI, m / z): [M+H]+ = 472.1.
[0637] Intermediate 3: 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)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0638] Step 1: preparation of tert-butyl 3-(7-chloro-8-fluoro-2-((1- (hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc Boc । ।
[0639] Intermediate 2-P1 (chiral column retention time: 2.248 min) (400 mg, 0.85 mmol) and cyclopropane-1,1-diyldimethanol (173 mg, 1.69 mmol) were dissolved in anhydrous acetonitrile (40 mL), and cesium carbonate (551 mg, 1.69 mmol) and triethylenediamine (28 mg, 0.25 mmol) were added. The mixture was reacted at 25 °C for 4 h. The reaction liquid was poured into a saturated aqueous ammonium chloride solution (150 mL), and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated to give tert-butyl 3-(7-chloro-8-fluoro-2-((1- (hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0640] LC-MS: (ESI, m / z): [M+H]+ = 538.2.
[0641] Step 2: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc OMOM
[0642] CataCXium A Pd-G3 (130 mg, 0.18 mmol) and cesium carbonate (875 mg, 2.68 mmol) were added to a mixed solution of tert-butyl 3-(7-chloro-8-fluoro-2-((1-(hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (482 mg, 0.90 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (917 mg, 1.79 mmol) in 1,4-dioxane (40 mL) and H2O (10 mL). The mixture was reacted at 85 °C for 18 h under nitrogen atmosphere. Water (10 mL) was added to the reaction liquid, and then the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0643] LC-MS: (ESI, m / z): [M+H]+ = 888.3.
[0644] Step 3: preparation of 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)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1- (hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (656 mg, 0.74 mmol) in dichloromethane (50 mL), and the mixture was reacted at room temperature for 3 h. The reaction liquid was poured into a saturated aqueous sodium carbonate solution (60 mL), and then the mixture was extracted with ethyl acetate (50 mL x 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated to give 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)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0646] LC-MS: (ESI, m / z): [M+H]+ = 886.3.
[0647] Intermediate 4: 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3- azaspiro[5.5]undecane-9-carbaldehyde Step 1: preparation of 3-azaspiro[5.5]undecane-9-carbaldehyde
[0648] tert-Butyl 9-formyl-3-azaspiro[5.5]undecane-3-carboxylate (400 mg, 1.42 mmol) was dissolved in DCM (4 mL), and then TFA (2 mL) was added. The mixture was stirred at room temperature for 1 h and concentrated to give 3-azaspiro[5.5]undecane-9-carbaldehyde, which was directly used in the next step.
[0649] Step 2: preparation of 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3- azaspiro[5.5]undecane-9-carbaldehyde
[0650] DIEA (720 mg, 5.61 mmol) and pentafluorophenyl 3-(2,4-dioxotetrahydropyrimidin-1(2 / / )-yl)-4-methoxybenzoate (641 mg, 1.49 mmol) were added to a solution of 3-azaspiro[5.5]undecane-9-carbaldehyde (500 mg, crude) in DMSO (10 mL), and the reaction liquid was stirred at room temperature for 1 h. Water was added to the reaction liquid, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give 3-(3-(2,4-dioxotetrahydropyrimidin-1(2 / )-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde.
[0651] LC-MS: (ESI, m / z): [M+H]+ = 428.3.
[0652] Intermediate 5: 3-(3-(2,4-dioxotetrahydropyrimidin-1(2 / )-yl)-4-methylbenzoyl)-3- azaspiro[5.5]undecane-9-carbaldehyde
[0653] The intermediate was prepared with reference to the preparation of intermediate 32 in the patent WO2023025159A1.
[0654] Intermediate 6: tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate
[0655] Step 1: preparation of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine O Cl CI-^y^N O CI'^^V^N^CI F H F
[0656] 7-Chloro-8-fluoro-1,2,3,4-tetrahydropyrido[4,3- ^]pyrimidine-2,4-dione (9.30 g, 43.14 mol) was dissolved in PCI3 (100 mL), and the mixture was cooled to 0 °C. DIEA (16.69 g, 129.42 mol) was slowly added dropwise with stirring. The reaction liquid was heated to 120 °C and reacted for 16 h. After the reaction was completed, the reaction liquid was concentrated, diluted with ethyl acetate, and then slowly poured into ice water to quench the reaction. The mixture was extracted with ethyl acetate (100 mL x 3), and the organic phase was separated. The organic phase was sequentially washed with a cooled aqueous NaHCO3 solution and saturated NaCl solution, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine.
[0657] Step 2: preparation of tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate were dissolved in THF (15 mL), and the mixture was cooled to -60 °C. A solution of tert -butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.71 g, 3.37 mmol) in THF (5 mL) was slowly added dropwise, and the mixture was reacted at -60 °C for 30 min. After the reaction was completed, the reaction liquid was poured into an aqueous NH4Cl solution (20 mL), and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated to give a crude product, which was purified by silica gel column chromatography (PE:EtOAc = 3:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0659] LC-MS: (ESI, m / z): [M+H]+ = 428.2.
[0660] Intermediate 7: preparation of 1-(2-methoxy-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0661] Step 1: preparation of tert-butyl 9-((4-((benzyloxy)carbonyl)piperazin-1-yl)methyl)-3- azaspiro[5.5]undecane-3-carboxylate Cbz \
[0662] AcOH (0.2 mL) was added to a mixed solution of tert-butyl 9-formyl-3-azaspiro[5.5]undecane-3-carboxylate (1.30 g, 4.62 mmol) and tert -butyl piperazine-1-carboxylate (1.82 g, 8.28 mmol) in DCM / MeOH (15 mL / 5 mL), and the mixture was stirred at room temperature for 0.5 h. NaBH(OAc)3 (2.25 g, 10.60 mmol) was then added in batches to the above mixture at 0-5 °C, and the mixture was stirred for another 0.5 h. The reaction liquid was directly concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (MeOH:DCM = 150:1 to 20:1) to give tert-butyl 9-((4-((benzyloxy)carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate.
[0663] LC-MS: (ESI, m / z): [M+H]+ = 486.3.
[0664] Step 2: preparation of benzyl 4-((3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate
[0665] HCl / 1,4-dioxane (6 N, 5 mL) was added to a solution of tert-butyl 9-((4-((benzyloxy)carbonyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylate (750 mg, 4.54 mmol) in 1,4-dioxane (5 mL). The mixture was stirred at room temperature for 1 h, and the reaction liquid was directly concentrated under reduced pressure to give benzyl 4-((3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate. The resulting crude product was directly used in the next step without purification.
[0666] Step 3: preparation of benzyl 4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4- methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate
[0667] Pentafluorophenyl 3-(2,4-dioxotetrahydropyrimidin-1(2 #)-yl)-4-methoxybenzoate (430 mg, 1.00 mmol) and DIEA (516 mg, 4.00 mmol) were added to a solution of benzyl 4-((3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate (632 mg, crude product) in DMSO (10 mL). The mixture was stirred at room temperature for 1 h and then purified by pre-HPLC to give benzyl 4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate.
[0668] LC-MS: (ESI, m / z): [M+H]+ = 632.3.
[0669] Step 4: preparation of 1-(2-methoxy-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3- carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0670] Pd / C (300 mg) was added to a solution of benzyl 4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2 H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate (586 mg, 0.93 mmol) in ethyl acetate (20 mL), and the mixture was stirred at room temperature for 16 h. The reaction liquid was filtered through celite, and the filtrate was concentrated under reduced pressure to give 1-(2-methoxy-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0671] LC-MS: (ESI, m / z): [M+H]+ = 498.2.
[0672] Intermediate 8: preparation of 1-(2-methyl-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3- carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0673] Step 1: preparation of benzyl 4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)- 3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate
[0674] Pentafluorophenyl 3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoate (921 mg, 2.3 mmol) and DIEA (995 mg, 7.7 mmol) were added to a solution of benzyl 4-((3-azaspiro[5.5]undecan-9- yl)methyl)piperazine-1-carboxylate hydrochloride (930 mg, 2.2 mmol) in DMSO (20 mL). The mixture was stirred at room temperature for 1 h and purified by pre-HPLC to give benzyl 4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1- carboxylate.
[0675] LC-MS: (ESI, m / z): [M+H]+ = 616.3.
[0676] Step 2: preparation of 1-(2-methyl-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3- carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0677] Pd / C (100 mg) was added to a solution of benzyl 4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2 H)-yl)- 4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazine-1-carboxylate (900 mg, 1.46 mmol) in ethyl acetate (20 mL), and the mixture was stirred at room temperature for 16 h. The reaction liquid was filtered through celite, and the filtrate was concentrated to give 1-(2-methyl-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0678] LC-MS: (ESI, m / z): [M+H]+ = 482.2.
[0679] Intermediate 9: tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4- yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0680] Step 1: preparation of tert-butyl 3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0681] tert-Butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (11.00 g, 51.57 mmol) and triethylamine (10.44 g, 103.14 mmol) were dissolved in acetonitrile (150 mL), and trityl chloride (21.57 g, 77.36 mmol) was added. The mixture was reacted at room temperature in the dark for 18 h. After the reaction was completed, petroleum ether (50 mL) was added to the reaction liquid, and the mixture was stirred for 30 min and filtered. The filter cake was stirred with water (100 mL) and filtered, and this procedure was repeated twice. The filter cake was then lyophilized to remove water, thus giving tert-butyl 3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0682] LC-MS: (ESI, m / z): [M+Na]+ = 477.2.
[0683] Step 2: preparation of tert-butyl 1-formyl-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0684] A solution of tert -butyl 3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (15 g, 32.9 mmol) in THF (150 mL) was cooled to -78 °C, and TMEDA (9.56 g, 82.2 mmol) and 5-BuLi (4.22 g, 65.8 mmol) were slowly added under nitrogen atmosphere. The mixture was stirred at -78 °C for 2 h. DMF (7.21 g, 98.7 mmol) was then added, and the mixture was warmed to room temperature and stirred for 16 h. The reaction liquid was poured into a saturated aqueous ammonium chloride solution (200 mL), and the mixture was extracted with ethyl acetate (200 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 10:1) to give tert-butyl 1-formyl-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0685] LC-MS: (ESI, m / z): [M+Na]+ = 505.2.
[0686] Step 3: preparation of intermediate 9-P1 and intermediate 9-P2
[0687] Methyltriphenylphosphonium bromide (18.22 g, 51.0 mmol) was dissolved in THF (80 mL), and t- BuOK (5.72 g, 51.0 mmol) was added to the mixture at 0 °C. After 10 min, tert-butyl 1-formyl-3-trityl-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (8.2 g, 17.0 mmol) was added at 0 °C, and then the reaction mixture was stirred at 25 °C for 18 h. The reaction liquid was poured into a saturated aqueous ammonium chloride solution (200 mL), and the mixture was extracted with ethyl acetate (200 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EtOAc = 50:1 to 5:1) to give tert-butyl 3-trityl-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate. The compound obtained from the purification was subjected to SFC resolution with the following method: chromatographic column: chiralpak-IG, 250 x 30 mm, 10 um; mobile phase: CO2 / MeOH (0.2% NH3 (7 M solution in MeOH)); gradient: 90 / 10, 6 min; flow rate: 80 g / min; wavelength: 214 nm; column temperature: 35 °C.
[0688] Intermediate 9-P1 (chiral column retention time: 1.357 min). SFC analysis method: (column: chiralpak-IG-3, 150 mm x 3 mm, 3 um); mobile phase: CO2 / MeOH (0.1% DEA); gradient: 90 / 10, 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm & 254 nm; column temperature: 37 °C; optical purity: 100%.
[0689] Intermediate 9-P2 (chiral column retention time: 1.744 min). SFC analysis method: (column: chiralpak-IG-3, 150 mm x 3 mm, 3 um); mobile phase: CO2 / MeOH (0.1% DEA); gradient: 90 / 10, 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm & 254 nm; column temperature: 37 °C; optical purity: 100%.
[0690]
[0691] LC-MS: (ESI, m / z): [M+Na]+ = 503.2. Step 4: preparation of tert-butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc Trt Boc intermediate 9-P1
[0692] added. Intermediate 9-P1 (1.50 g, 3.11 mmol) was dissolved in DCM (20 mL), and AcOH (10 mL) was The mixture was reacted at 45 °C for 4 h. The reaction liquid was directly concentrated, and the resulting crude product was purified by silica gel column chromatography (DCM:MeOH (NH3) = 10:1) to give tert-butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0693]
[0694] LC-MS: (ESI, m / z): [M-56+H]+ = 183.2. Step 5: preparation of tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8- diazabicyclo[3.2.1]octane-8-carboxylate 1.70 mmol) were dissolved in tetrahydrofuran (6 mL), and the mixture was cooled to -50 °C. tert-Butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (367 mg, 1.53 mmol) was added, and the mixture was reacted at -50 °C for 3 h. The reaction liquid was poured into a saturated aqueous ammonium chloride solution (10 mL), and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0696] LC-MS: (ESI, m / z): [M+H]+ = 454.1.
[0697] Step 6: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 3-(2,7-dichloro-8-fluoropyrido[4,3- ^]pyrimidin-4-yl)-1-vinyl-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (450 mg, 0.99 mmol) and (R)-3-(4-benzylpiperazin-1-yl)-2- methylpropan-1-ol (370 mg, 1.48 mmol) were dissolved in THF (8 mL), and the mixture was added to a sealed tube reactor. Cs2CO3 (966 mg, 2.96 mmol) was then added. The mixture was reacted at 90 °C for 16 h. After the reaction was completed, the reaction liquid was cooled to room temperature. Water (10 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH = 15:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0699] LC-MS: (ESI, m / z): [M+H]+ = 666.3.
[0700] Step 7: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7- (7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)- 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0701] CataCXium A Pd-G3 (109 mg, 0.15 mmol) and cesium carbonate (731 mg, 2.24 mmol) were added to a mixed solution of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (500 mg, 0.75 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (460 mg, 0.90 mmol) in 1,4-dioxane (5 mL) and H2O (1 mL). The mixture was reacted at 85 °C for 16 h under nitrogen atmosphere. Water (15 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (15 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 15:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0702] LC-MS: (ESI, m / z): [M / 2+H]+ = 508.9.
[0703] Step 8: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0704] tert-Butyl 3-(2-((R )-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (490 mg, 0.48 mmol) was dissolved in DCM (5 mL), and 1-chloroethyl chloroformate (206 mg, 1.44 mmol) and DIEA (124 mg, 0.96 mmol) were added. The mixture was reacted at room temperature for 1 h. The reaction liquid was directly concentrated under reduced pressure. MeOH (5 mL) was added to the resulting concentrate, and then the mixture was heated to 50 °C and reacted for 10 min. After the reaction was completed, the mixture was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0705] LC-MS: (ESI, m / z): [M / 2+H]+ = 463.8.
[0706] Intermediate 10: tert-butyl 1-ethyl-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0707] Step 1: preparation of tert-butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0708] Intermediate 9-P1 (1.50 g, 3.11 mmol) was dissolved in DCM (20 mL), and AcOH (10 mL) was added. The mixture was reacted at 45 °C for 4 h. The reaction liquid was directly concentrated, and the resulting crude product was purified by silica gel column chromatography (DCM:MeOH (NH3) = 10:1) to give tert-butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0709] LC-MS: (ESI, m / z): [M-56+H]+ = 183.2.
[0710] Step 2: preparation of tert-butyl 1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc Boc i i tr er H H
[0711] tert -Butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (330 mg, 1.38 mmol) was dissolved in MeOH (3 mL), and Pd / C (60 mg) was added. The mixture was reacted at room temperature for 3 h under hydrogen atmosphere. After the reaction was completed, the mixture was filtered and concentrated to give a crude product of tert-butyl 1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0712] LC-MS: (ESI, m / z): [M+H]+ = 241.3.
[0713] Step 3: preparation of tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-ethyl-3,8- diazabicyclo[3.2.1]octane-8-carboxylate Boc
[0714] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (292 mg, 1.16 mmol) and triethylamine (351 mg, 3.47 mmol) were dissolved in tetrahydrofuran (6 mL), and the mixture was cooled to -50 °C. tert-Butyl 1- ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (250 mg, 1.04 mmol) was added, and the mixture was reacted at -50 °C for 3 h. The reaction liquid was poured into a saturated aqueous ammonium chloride solution (10 mL), and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 3:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0715] LC-MS: (ESI, m / z): [M+H]+ = 456.2.
[0716] Step 4: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro- 8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidm-4-yl)-1-ethyl-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (280 mg, 0.61 mmol) and (R)-3-(4-benzylpiperazin-1-yl)-2- methylpropan-1-ol (305 mg, 1.23 mmol) were dissolved in THF (3 mL), and the mixture was added to a sealed tube reactor. Cs2CO3 (600 mg, 1.84 mmol) was then added, and the mixture was reacted at 90 °C for 16 h. After the reaction liquid was cooled to room temperature, water (10 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tertbutyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0718] LC-MS: (ESI, m / z): [M+H]+ = 668.3.
[0719] Step 5: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7- (7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)- 1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate to a mixed solution of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8- fluoropyrido[4,3-d]pyrimidin-4-yl)-1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (240 mg, 0.36 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1- yl)ethynyl)triisopropylsilane (239 mg, 0.47 mmol) in 1,4-dioxane (3 mL) and H2O (0.6 mL). The mixture was reacted at 85 °C for 16 h under nitrogen atmosphere. Water (15 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (15 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0721] LC-MS: (ESI, m / z): [M / 2+H]+ = 509.9.
[0722] Step 6: preparation of tert-butyl 1-ethyl-3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0723] tert-Butyl 3-(2-((R )-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-ethyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (255 mg, 0.25 mmol) was dissolved in DCM (2 mL), and 1- chloroethyl chloroformate (107 mg, 0.75 mmol) and DIEA (97 mg, 0.75 mmol) were added. The mixture was reacted at room temperature for 1 h and concentrated under reduced pressure. MeOH (3 mL) was added to the concentrate, and then the mixture was heated to 50 °C and reacted for 10 min. After the reaction was completed, the mixture was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 15:1) to give tert-butyl 1-ethyl-3-(8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0724] LC-MS: (ESI, m / z): [M / 2+H]+ = 464.9.
[0725] Intermediate 11: intermediate 11-P1 and intermediate 11-P2
[0726] Step 1: preparation of tert-butyl 3-benzyl-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate
[0727] tert-Butyl 3-benzyl-1-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (2.5 g, 7.53 mmol) was dissolved in acetonitrile (30 mL), and silver oxide (5.24 g, 22.59 mmol) and CD3I (1.64 g, 11.30 mmol) were added. The mixture was reacted at room temperature in the dark for 72 h. After the reaction was completed, the reaction liquid was filtered and purified by silica gel column chromatography (PE:EA = 20:1 to 3:1) to give tert-butyl 3-benzyl-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0728] LC-MS: (ESI, m / z): [M+H]+ = 350.2.
[0729] Step 2: preparation of tert-butyl 1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0730] tert-Butyl 3-benzyl-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (450 mg, 1.29 mmol) was dissolved in methanol (6 mL), and Pd / C (300 mg) was added. The mixture was reacted at 25 °C for 16 h. The reaction liquid was filtered, and the filtrate was concentrated to give tert-butyl 1- ((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0731] LC-MS: (ESI, m / z): [M-Boc+H]+ = 160.3.
[0732] Step 3: preparation of intermediate 11-P1 and intermediate 11-P2 Boc Cl F Boc OCD3 Cl intermediate 11-P1 or intermediate 11-P2 intermediate 11-P2 or intermediate 11-P1
[0733] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (353 mg, 1.40 mmol) and triethylamine (424 mg, 4.19 mmol) were dissolved in tetrahydrofuran (6 mL), and the mixture was cooled to -78 °C. tert-Butyl 1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (326 mg, 1.26 mmol) was added, and the mixture was reacted at -78 °C for 3 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (10 mL), and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate. The compound obtained from the purification was subjected to SFC resolution with the following method: chromatographic column: Daicel Chiralcel OD, 250 x 30 mm, 10 um; mobile phase: CO2 / MeOH (0.2% NH3 (7 M solution in MeOH) = 80 / 20; flow rate: 150 mL / min; wavelength: 214 nm; column temperature: 35 °C.
[0734] Intermediate 11-P1 (chiral column retention time: 1.408 min). SFC analysis method: column: Daicel chiralpak OD_3, 3 x 150 mm, 3 um; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 80 / 20, 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0735] Intermediate 11-P2 (chiral column retention time: 1.753 min). SFC analysis method: column: Daicel chiralpak OD_3, 3 x 150 mm, 3 um; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 80 / 20, 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0736] LC-MS: (ESI, m / z): [M+H]+ = 475.2.
[0737] Intermediate 12: tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0738] Step 1: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro- 8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc £r° N intermediate 2-P1
[0739] Intermediate 2-P1 (chiral column retention time: 2.248 min) (400 mg, 0.85 mmol) and (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-1-ol (421 mg, 1.69 mmol) were dissolved in THF (5 mL), and the mixture was added to a sealed tube reactor. Cs2CO3 (828 mg, 2.54 mmol) was then added. The mixture was reacted at 90 °C for 16 h. The reaction liquid was cooled to room temperature, and water (10 mL) was added. The mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 1:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0740] LC-MS: (ESI, m / z): [M+H]+ = 684.3.
[0741] Step 2: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0742] CataCXium A Pd-G3 (85 mg, 0.12 mmol) and cesium carbonate (571 mg, 1.75 mmol) were added to a mixed solution of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8- fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (400 mg, 0.58 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (360 mg, 0.7 mmol) in 1,4-dioxane (3 mL) and H2O (0.6 mL). The mixture was reacted at 85 °C for 16 h under nitrogen atmosphere. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2- methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0743] LC-MS: (ESI, m / z): [M / 2+H]+ = 517.9.
[0744] Step 3: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3- d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0745] tert -Butyl 3 -(2-((R )-3 -(4-benzylpiperazin-1 -yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1- (methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (510 g, 0.49 mmol) was dissolved in DCM (5 mL), and then 1-chloroethyl chloroformate (211 mg, 1.48 mmol) and N,N-diisopropylethylamine (191 mg, 1.48 mmol) were added. The mixture was reacted at room temperature for 1 h and concentrated under reduced pressure. The concentrate was dissolved in MeOH (5 mL), and the resulting mixture was heated to 50 °C and reacted for 10 min. After the reaction was completed, the mixture was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0746] LC-MS: (ESI, m / z): [M / 2+H]+ = 472.8.
[0747] Intermediate 13: 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 Boc OMOM
[0748] The intermediate was prepared with reference to the preparation method for intermediate 17 in WO2024083258A1.
[0749] Intermediate 14: intermediate 14-P1 and intermediate 14-P2
[0750] Step 1: preparation of tert-butyl 1-methyl-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0751] tert-Butyl 3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (5.0 g, 10.99 mmol) was dissolved in tetrahydrofuran (30 mL). TMEDA (2.56 g, 21.99 mmol) was added, and then 5-BuLi (1.41 g, 21.99 mmol) was added dropwise at -30 °C. After the dropwise addition was completed, the mixture was stirred at 0 °C for 30 min, and iodomethane (4.68 g, 31.99 mmol) was added dropwise. The mixture was then stirred at 0 °C for another 15 min. After the reaction was completed, a saturated aqueous ammonium chloride solution (30 mL) was added to quench the reaction, and then the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EtOAc = 100:1 to 10:1) to give tert-butyl 1-methyl-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0752] LC-MS: (ESI, m / z): [M+Na]+ = 491.1.
[0753] Step 2: preparation of tert-butyl 1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0754] tert-Butyl 1-methyl-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (3 g, 6.40 mmol) was dissolved in dichloromethane (15 mL), and glacial acetic acid (15 mL) was added. The mixture was refluxed at 50 °C for 1 h. After the reaction was completed, the reaction liquid was directly concentrated, and the resulting crude product was purified by silica gel column chromatography (DCM:MeOH (NH3) = 20:1) to give tert-butyl 1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0755] LC-MS: (ESI, m / z): [M+H]+ = 227.2.
[0756] Step 3: preparation of intermediate 14-P1 and intermediate 14-P2
[0757] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (1.5 g, 5.94 mmol) and triethylamine (1.8 g, 17.83 mmol) were dissolved in tetrahydrofuran (20 mL), and the mixture was cooled to -50 °C. tert-Butyl 1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.2 g, 5.30 mmol) was added, and the mixture was reacted at -50 °C for 3 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (50 mL), and then the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8- diazabicyclo[3.2.1]octane-8-carboxylate. The racemate obtained from the purification was subjected to chiral SFC resolution with the following method: chromatographic column: Daicel CHIRALCEL IB-N, 250 mm x 30 mm, 10 um; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 70 / 30, 6 min; flow rate: 80 g / min; wavelength: 214 nm; column temperature: 35 °C.
[0758] Intermediate 14-P1 (chiral column retention time: 1.252 min). SFC analysis method: column: Daicel CHIRALPAK OD_3, 3 x 150 mm, 3 pm; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 70 / 30, 6 min; flow rate: 2 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0759] Intermediate 14-P2 (chiral column retention time: 1.663 min). SFC analysis method: column: Daicel CHIRALPAK OD_3, 3 x 150 mm, 3 pm; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 70 / 30, 6 min; flow rate: 2 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0760] Confirmation of absolute configuration of intermediate 14-P1:
[0761] Step 4: preparation of methyl (R)-2-methyl-5-oxopyrrolidine-2-carboxylate
[0762] (R )-2-Methyl-5-oxopyrrolidine-2-carboxylic acid (500 mg, 3.50 mmol) was dissolved in DMF (5 mL), and potassium carbonate (966 mg, 7.0 mmol) was added. The mixture was reacted at room temperature for 2 h. After the reaction was completed, the reaction liquid was poured into an aqueous solution (20 mL), and the mixture was extracted with ethyl acetate (3 x 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated to obtain a crude product, which was subjected to silica gel column chromatography (PE / EA = 20 / 1 to 10 / 1) to give methyl (R)-2-methyl-5-oxopyrrolidine-2-carboxylate.
[0763] LC-MS: (ESI, m / z): [M+H]+ = 158.2.
[0764] Step 5: preparation of methyl (R)-1-benzyl-2-methyl-5-oxopyrrolidine-2-carboxylate
[0765] Methyl (R )-2-methyl-5-oxopyrrolidine-2-carboxylate (300 mg, 1.91 mmol) was dissolved in DMF (5 mL), and the mixture was cooled to 0 °C. NaH (90 mg) was added, and the mixture was stirred at 0 °C for 30 min. Benzyl bromide (488 mg, 2.87 mmol) was then added, and the mixture was stirred at 25 °C for 30 min. After the reaction was completed, a saturated aqueous ammonium chloride solution (1 mL) was added dropwise to quench the reaction, and water (20 mL) was added. The mixture was extracted with ethyl acetate (3 x 20 mL). The organic phases were combined, washed with a saturated NaCl solution (20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated to give a crude product, which was subjected to silica gel column chromatography (DCM / MeOH = 100 / 1) to give methyl (R)-1-benzyl-2-methyl-5-oxopyrrolidine-2-carboxylate.
[0766] LC-MS: (ESI, m / z): [M+H]+ = 248.2.
[0767] Step 6: preparation of (R)-1-benzyl-5-ethoxy-2-(methoxycarbonyl)-2-methyl-3,4-dihydro-2H-pyrrol-1-ium tetrafluoroborate
[0768] Methyl (R)-1-benzyl-2-methyl-5-oxopyrrolidine-2-carboxylate (360 mg, 1.46 mmol) was dissolved in dichloromethane (5 mL), and triethyloxonium tetrafluoroborate (554.8 mg, 2.92 mmol) was added at 0 °C. The mixture was reacted at room temperature for 24 h. After the reaction was completed, the reaction liquid was concentrated to give a crude product of (R)-1-benzyl-5-ethoxy-2-(methoxycarbonyl)-2-methyl-3,4-dihydro-2H-pyrrol-1-ium tetrafluoroborate.
[0769] LC-MS: (ESI, m / z): [M]+ = 276.2.
[0770] Step 7: preparation of methyl (R)-1-benzyl-2-methyl-5-(nitromethylene)pyrrolidine-2-carboxylate
[0771] The crude product of (R)-1-benzyl-5-ethoxy-2-(methoxycarbonyl)-2-methyl-3,4-dihydro-2H-pyrrol-1-ium tetrafluoroborate (900 mg, 0.88 mmol), was dissolved in DCM (5 mL), and nitromethane (270 mg, 4.4 mmol) and triethylamine (355.5 mg, 3.52 mmol) were added in an ice bath. The mixture was reacted at room temperature for 2 h. After the reaction was completed, the reaction liquid was concentrated to give a crude product, which was subjected to silica gel column chromatography (DCM / MeOH = 50 / 1) to give methyl (R)-1-benzyl-2-methyl-5-(nitromethylene)pyrrolidine-2-carboxylate.
[0772] LC-MS: (ESI, m / z): [M+H]+ = 291.1.
[0773] Step 8: preparation of (1R,5S)-1-methyl-3,8-diazabicyclo[3.2.1]octan-2-one
[0774] Methyl (R)-1-benzyl-2-methyl-5-(nitromethylene)pyrrolidine-2-carboxylate (90 mg, 0.31 mmol) was dissolved in MeOH (5 mL), and palladium on carbon (50 mg) was added at room temperature. The mixture was purged with hydrogen gas three times using a hydrogen balloon and reacted at room temperature for 16 h. After the reaction was completed, the palladium on carbon was removed by filtration, and the filtrate was concentrated to give (1R,5S)-1-methyl-3,8-diazabicyclo[3.2.1]octan-2-one.
[0775] LC-MS: (ESI, m / z): [M+H]+ = 141.1 .
[0776] Step 9: preparation of tert-butyl (1R,5S)-1-methyl-2-oxo-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0777] The crude product (1R ,5 5)-1-methyl-3,8-diazabicyclo[3.2.1]octan-2-one (40 mg, 0.31 mmol) was dissolved in MeOH (3 mL), and di-tert-butyl dicarbonate (134 mg, 0.62 mmol) was added at room temperature. The mixture was reacted at room temperature for 2 h. After the reaction was completed, the reaction liquid was concentrated to give a crude product, which was subjected to silica gel column chromatography (DCM / MeOH = 20 / 1) to give tert-butyl (1R,5S)-1-methyl-2-oxo-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0778] LC-MS: (ESI, m / z): [M-56+H]+ = 185.2 .
[0779] Step 10: preparation of tert-butyl (1R,5S)-3-benzyl-1-methyl-2-oxo-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0780] tert-Butyl (1R,55)-1-methyl-2-oxo-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30 mg, 0.125 mmol) was dissolved in DMF (3 mL), and the mixture was cooled to 0 °C. NaH (10 mg) was added, and the mixture was stirred at 0 °C for 30 min. Benzyl bromide (32.3 mg, 0.19 mmol) was then added, and the mixture was stirred at 25 °C for 30 min. After the reaction was completed, a saturated aqueous ammonium chloride solution (0.5 mL) was added dropwise to quench the reaction, and water (10 mL) was added. The mixture was extracted with ethyl acetate (3 x 10 mL). The organic phases were combined, washed with a saturated NaCl solution (20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated to give a crude product, which was subjected to silica gel column chromatography (PE / EA = 20 / 1) to give tert-butyl (1R,5S)-3-benzyl-1-methyl-2-oxo-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0781] LC-MS: (ESI, m / z): [M-56+H]+ = 275.1.
[0782] Step 11: preparation of tert-butyl (1R,5S)-3-benzyl-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0783] tert-Butyl (1R,55)-3-benzyl-1-methyl-2-oxo-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30 mg, 0.09 mmol) was dissolved in THF (3 mL), and 10 M borane dimethyl sulfide complex (0.18 mL, 1.8 mmol) was added at room temperature. The mixture was reacted at 50 °C for 2 h. After the reaction was completed, the reaction liquid was concentrated to give a crude product, which was subjected to silica gel column chromatography (PE / EA = 20 / 1) to give tert-butyl (1R,5S)-3-benzyl-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0784] LC-MS: (ESI, m / z): [M+H]+ = 317.2.
[0785] Step 12: preparation of tert-butyl (1R,5S)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate B°c Boc zN^ N Bn H
[0786] tert-Butyl (1R ,5 5)-3-benzyl-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (20 mg, 0.063 mmol) was dissolved in MeOH (3 mL), and palladium on carbon (20 mg) was added at room temperature. The mixture was purged with hydrogen gas three times using a hydrogen balloon and reacted at room temperature for 3 h. After the reaction was completed, the palladium on carbon was removed by filtration, and the filtrate was concentrated to give a crude product, which was subjected to silica gel column chromatography (DCM / 4 M NH3 in MeOH = 30 / 1) to give tert-butyl (1R,5S)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0787] LC-MS: (ESI, m / z): [M-56+H]+ = 171.2.
[0788] 1H NMR (400 MHz, CDCl3) 5 4.20 (s, 1H), 2.99 (d, J = 10.8 Hz, 2H), 2.56 (dd, J = 32.4, 12.0 Hz, 2H), 2.01 - 1.92 (m, 2H), 1.74 - 1.65 (m, 2H), 1.47 (s, 9H), 1.43 (s, 3H).
[0789] Step 13: preparation of tert-butyl (1R,5S)-3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1- methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0790] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (15 mg, 0.059 mmol) and triethylamine (18 mg, 0.178 mmol) were dissolved in tetrahydrofuran (2 mL), and the mixture was cooled to -50 °C. tert -Butyl (1R,5S)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (12 mg, 0.053 mmol) was added, and the mixture was reacted at -50 °C for 3 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (5 mL), and then the mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl (1R,5S)-3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0791] Chiral analysis method: column: Daicel CHIRALPAK OD_3, 3 x 150 mm, 3 um: mobile phase: CO2 / MeOH (0.1% DEA); gradient: 70 / 30, 6 min; flow rate: 2 mL / min; wavelength: 214 nm; column temperature: 37 °C.
[0792] The chiral column retention time of tert-butyl (1R,5S)-3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate was the same as that of intermediate 14-P1 from the chiral resolution in step 3. By the method of chiral synthesis, the absolute configuration of intermediate 14-P1 was determined to be (1R,5S).
[0793] Intermediate 15: 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)-1-vinyl-5Z3-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0794] Step 1: preparation of tert-butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc Trt Boc H intermediate 9-P1
[0795] Intermediate 9-P1 (1.50 g, 3.11 mmol) was dissolved in DCM (20 mL), and AcOH (10 mL) was added. The mixture was reacted at 45 °C for 4 h. The reaction liquid was directly concentrated, and the resulting crude product was purified by silica gel column chromatography (DCM:MeOH (NH3) = 10:1) to give tert-butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0796] LC-MS: (ESI, m / z): [M-56+H]+ = 183.2.
[0797] Step 2: preparation of tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d] pyrimidin-4-yl)-1-vinyl-5Z3- 3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0798] Triethylamine (481 mg, 4.75 mmol) and tert-butyl 1-vinyl-3,8-diazabicyclo[3.2.1]octane-8- carboxylate (341 mg, 1.43 mmol) were added to a solution of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (400 mg, 1.58 mmol) in tetrahydrofuran (6 mL) at -50 °C, and the mixture was stirred at -50 °C for 3 h. After the reaction was completed, a saturated aqueous ammonium chloride solution (20 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give a crude product, which was subjected to silica gel column chromatography (PE:EA = 3:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3- d] pyrimidin-4-yl)-1-vinyl-5Z3-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0799] LC-MS: (ESI, m / z): [M+H]+ = 454.1.
[0800] Step 3: preparation of tert-butyl 3-(7-chloro-8-fluoro-2-((1- (hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-1-vinyl-5Z3-3,8- diazabicyclo[3.2.1]octane-8-carboxylate Boc ?oc
[0801] Triethylenediamine (57 mg, 0.51 mmol) and anhydrous cesium carbonate (987 mg, 3.03 mmol) were added to a solution of tert -butyl 3-(2,7-dichloro-8-fluoropyrido[4,3- d] pyrimidin-4-yl)-1-vmyl-5Z3-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (460 mg, 1.01 mmol) and cyclopropane-1,1-diyldimethanol (206 mg, 2.02 mmol) in anhydrous acetonitrile (5 mL), and the mixture was reacted at 25 °C for 2 h. After the reaction was completed, the reaction liquid was filtered and concentrated to give a crude product, which was subjected to silica gel column chromatography (PE:EA = 3:1) to give tert-butyl 3-(7-chloro-8-fluoro-2-((1-(hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-1-vinyl-5Z3-3,8- diazabicyclo[3.2.1]octane-8-carboxylate.
[0802] LC-MS: (ESI, m / z): [M+H]+ = 520.2.
[0803] Step 4: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-5Z3-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0804] were (hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3- d]pyrimidin-4-yl)-1-vinyl-5Z3-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (364 mg, 0.70 mmol) and ((2-fluoro-6-(methoxymethoxy)-8- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (430 mg, 0.84 mmol) in 1,4-dioxane (5 mL) and water (1 mL). The mixture was purged with nitrogen, and then the mixture was stirred at 85 °C for 16 h under nitrogen atmosphere. After the reaction was completed, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give a crude product, which was purified by silica gel column chromatography (PE:EA = 1:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3- ^]pyrimidin-4-yl)-1-vinyl-5Z3-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0805] LC-MS: (ESI, m / z): [M+H]+ = 870.3.
[0806] Step 5: preparation of 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)-1-vinyl-5Z3 -3,8 -diazabicyclo[3.2.1]octane-8-carboxylate
[0807] A solution of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-vinyl-5Z3-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (460 mg, 0.53 mmol) in dichloromethane (3 mL) was cooled to 0 °C from room temperature, and Dess-Martin reagent (448 mg, 1.06 mmol) was added. The mixture was stirred at 25 °C for 2 h. After the reaction was completed, a saturated aqueous sodium thiosulfate solution (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL x 3). The organic phases were combined, washed with a saturated sodium bicarbonate solution and a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the resulting crude product was purified by silica gel column chromatography (PE:EA = 1:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((1-formylcyclopropyl)methoxy)pyrido[4,3-t / ]pynmidin-4-yl)-1 -vinyl-5Z3-3,8-diazabicyclo[3.2.1 ]octane-8-carboxylate.
[0808] LC-MS: (ESI, m / z): [M+H]+ = 868.4.
[0809] Intermediate 16: tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc OMOM
[0810] The intermediate was prepared with reference to the preparation method for intermediate 25 in WO2024083258A1.
[0811] Intermediate 17: 9-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-1-oxa-9- azaspiro[5.5]undecane-3-carbaldehyde
[0812] DIEA (6.84 g, 52.95 mmol) and pentafluorophenyl 3-(2,4-dioxotetrahydropyrimidin-1(2 / / )-yl)-4-methoxybenzoate (4.39 g, 10.59 mmol) were added to a solution of 1-oxa-9-azaspiro[5.5]undecane-3-carbaldehyde (1.94 g, crude product) in DMF (20 mL), and the reaction liquid was stirred at room temperature for 1 h. Water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to give 9-(3-(2,4-dioxotetrahydropyrimidin-1(2 / )-yl)-4-methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecane-3-carbaldehyde.
[0813] LC-MS (ESI) m / z: [M+H]+ = 430.2.
[0814] Intermediate 18: 3-(3-(2,4-dioxotetrahydropyrimidin-1(2 / )-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde
[0815] The intermediate was prepared with reference to the preparation method for intermediate 3 in WO2024083258A1.
[0816] Intermediate 19: preparation of 1-(2-methoxy-5-(3-(piperazin-1-ylmethyl)-3Z3-1-oxa-9- azaspiro[5.5]undecane-9-carbonyl)phenyl)dihydropyrimidine-2,4(1 / ,3 / )-dione
[0817] Step 1: preparation of tert-butyl 4-allyl-4-((2-(methoxycarbonyl)allyl)oxy)piperidine-1-carboxylate
[0818] tert-Butyl 4-allyl-4-hydroxypiperidine-1-carboxylate (9.0 g, 37.29 mmol) was dissolved in DMF (200 mL), and NaH (1.79 g, 44.75 mmol) was added at 0 °C. The mixture was reacted at room temperature for 1 h and then cooled to 0 °C. Methyl 2-(bromomethyl)acrylate (6.68 g, 37.29 mmol) was then added, and the mixture was reacted at room temperature for 72 h. After the reaction was completed, NH4Cl (200 mL) was added, and the mixture was extracted with ethyl acetate (200 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 5:1) to give tert-butyl 4-allyl-4-((2-(methoxycarbonyl)allyl)oxy)piperidine-1-carboxylate.
[0819] LC-MS: (ESI, m / z): [M+Na]+ = 362.2
[0820] Step 2: preparation of 9-(tert-butyl) 3-methyl 1-oxa-9-azaspiro[5.5]undec-3-ene-3,9-dicarboxylate
[0821] tert-Butyl 4-allyl-4-((2-(methoxycarbonyl)allyl)oxy)piperidine-1-carboxylate (3.0 g, 8.84 mmol) was dissolved in DCE (700 mL), and Grubbs II catalyst (229 mg, 0.27 mmol) was added. The mixture was reacted at 90 °C for 16 h. The reaction liquid was cooled to room temperature and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 5:1) to give 9-(tert-butyl) 3-methyl 1-oxa-9-azaspiro[5.5]undec-3-ene-3,9-dicarboxylate.
[0822] LC-MS: (ESI, m / z): [M+Na]+ = 334.2
[0823] Step 3: preparation of 9-(tert-butyl) 3-methyl 1-oxa-9-azaspiro[5.5]undecane-3,9-dicarboxylate
[0824] Pd(OH)2 (586 mg, 4.17 mmol) was added to a solution of 9-(tert-butyl) 3-methyl 1-oxa-9- azaspiro[5.5]undec-3-ene-3,9-dicarboxylate (1.3 g, 4.17 mmol) in EtOH (20 mL), and the mixture was reacted at 50 °C for 16 h under H2 atmosphere. The reaction liquid was cooled to room temperature and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 5:1) to give 9-(tert-butyl) 3-methyl 1-oxa-9-azaspiro[5.5]undecane-3,9-dicarboxylate.
[0825] Step 4: preparation of methyl 1-oxa-9-azaspiro[5.5]undecane-3-carboxylate
[0826] Hydrochloric acid / 1,4-dioxane (4.0 M, 50 mL) was added to a solution of 9-( tert-butyl) 3-methyl 1-oxa-9-azaspiro[5.5]undecane-3,9-dicarboxylate (1.386 g, 44.10 mmol) in dichloromethane (100 mL) at 0 °C, and the mixture was stirred at 25 °C for 2 h. The reaction liquid was directly concentrated under reduced pressure at 40 °C to give methyl 1-oxa-9-azaspiro[5.5]undecane-3-carboxylate, which was directly used in the next step.
[0827] LC-MS: (ESI, m / z): [M+H]+ = 214.1.
[0828] Step 5: preparation of methyl 9-benzyl-1-oxa-9-azaspiro[5.5]undecane-3-carboxylate
[0829] Benzaldehyde (6.534 g, 61.57 mmol) was added to a solution of methyl 1-oxa-9-azaspiro[5.5]undecane-3-carboxylate (8.755 g, crude product) in dichloromethane (180 mL). The mixture was reacted at room temperature for 2 h. Sodium triacetoxyborohydride (17.400 g, 82.10 mmol) was then added, and the mixture was reacted at room temperature for another 16 h. The reaction liquid was poured into a saturated aqueous ammonium chloride solution (200 mL), and then the mixture was extracted with dichloromethane (150 mL x 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The concentrate was stirred with a mixed solvent of petroleum ether / dichloromethane (10 / 1, 200 mL), and the resulting mixture was filtered and dried to give methyl 9-benzyl-1-oxa-9-azaspiro[5.5]undecane-3-carboxylate.
[0830] LC-MS: (ESI, m / z): [M+H]+ = 304.3.
[0831] Step 6: preparation of (9-benzyl-1-oxa-9-azaspiro[5.5]undecan-3-yl)methanol
[0832] Lithium aluminum hydride / tetrahydrofuran (1.0 M, 37.3 mL, 37.33 mmol) was added to a solution of methyl 9-benzyl-1-oxa-9-azaspiro[5.5]undecane-3-carboxylate (11.363 g, 37.33 mmol) in tetrahydrofuran (200 mL) at 0 °C. The mixture was stirred at 0-25 °C for 2 h. After the reaction was completed, water (1.4 mL), an aqueous sodium hydroxide solution (15% (w / w), 1.4 mL), and water (1.4 mL) were added sequentially at 10-minute intervals. The mixture was stirred for 30 min. Anhydrous sodium sulfate (10 g) was then added, and the mixture was stirred for another 30 min and filtered. The filtrate was concentrated under reduced pressure to give (9-benzyl-1-oxa-9-azaspiro[5.5]undecan-3-yl)methanol.
[0833] LC-MS: (ESI, m / z): [M+H]+ = 276.2.
[0834] Step 7: preparation of 9-benzyl-1-oxa-9-azaspiro[5.5]undecane-3-carbaldehyde
[0835] Dess-Martin oxidant (22.507 g, 53.06 mmol) was added to a solution of (9-benzyl-1-oxa-9-azaspiro[5.5]undecan-3-yl)methanol (9.778 g, 35.38 mmol) in dichloromethane (180 mL) at 0 °C. The mixture was stirred at 25 °C for 4 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous sodium carbonate solution (250 mL), and the mixture was extracted with ethyl acetate (200 mL x 2). The organic phases were combined, washed with a saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 9-benzyl-1-oxa-9-azaspiro[5.5]undecane-3-carbaldehyde.
[0836] LC-MS: (ESI, m / z): [M+H]+ = 274.2.
[0837] Step 8: preparation of 19-P1 and 19-P2 19-P1 or19-P2 19-P2 or 19-P1
[0838] tert-Butyl piperazine-1-carboxylate (9.870 g, 52.71 mmol) was added to a solution of 9-benzyl-1-oxa-9-azaspiro[5.5]undecane-3-carbaldehyde (9.641 g, 35.14 mmol) in dichloromethane (180 mL). The mixture was reacted at room temperature for 2 h. Sodium triacetoxyborohydride (14.894 g, 70.28 mmol) was then added, and the mixture was reacted at room temperature for another 16 h. The reaction liquid was poured into a saturated aqueous ammonium chloride solution (500 mL), and the mixture was extracted with ethyl acetate (200 mL x 2). The organic phase was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 20:1) to give tert-butyl 4-((9-benzyl-1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate. The compound obtained from the purification was subjected to SFC resolution with the following method: chromatographic column: Daicel chiralpak IG, 30 x 250 mm, 10 jam; mobile phase: COa / MeOH (0.2% NH3 (7 M solution in MeOH)); gradient: 65 / 35, 6 min; flow rate: 100 mL / min; wavelength: 214 nm; column temperature: 35 °C.
[0839] 19-P1 (chiral column retention time: 2.513 min). SFC analysis method: column: Daicel chiralpak AD_3, 3 x 150 mm, 3 am; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 70 / 30, 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0840] 1H NMR (400 MHz, CD3OD) 5 7.39 - 7.36 (m, 5H), 3.85 (s, 2H), 3.78 - 3.73 (m, 1H), 3.50 - 3.35 (m, 4H), 3.31 - 3.27 (m, 4H), 2.90 - 2.60 (m, 4H), 2.40 - 2.25 (m, 4H), 2.25 - 2.15 (m, 2H), 1.85 - 1.50 (m, 6H), 1.45 (s, 9H).
[0841] LC-MS: (ESI, m / z): [M+H]+ = 444.3.
[0842] 19-P2 (chiral column retention time: 3.171 min). SFC analysis method: column: Daicel chiralpak AD_3, 3 x 150 mm, 3 am; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 70 / 30, 6 min; flow rate: 2.0 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0843] 1H NMR (400 MHz, CD3OD) 5 7.52 - 7.46 (m, 5H), 4.23 (s, 2H), 3.81 - 3.77 (m, 1H), 3.50 - 3.35 (m, 4H), 3.35 - 3.27 (m, 4H), 3.25 - 3.00 (m, 4H), 2.40 - 2.35 (m, 4H), 2.35 - 2.15 (m, 2H), 1.85 - 1.55 (m, 6H), 1.45 (s, 9H).
[0844] LC-MS: (ESI, m / z): [M+H]+ = 444.3.
[0845] Step 9: preparation of tert-butyl 4-((1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1- carboxylate
[0846] Pd / C (0.20 g) was added to a solution of 19-P1 (1.10 g, 2.50 mmol) in methanol (10 mL), and the mixture was purged with hydrogen gas three times using a hydrogen balloon and stirred at 25 °C for 16 h. The reaction liquid was filtered through celite, and the filtrate was concentrated to give a crude product of tertbutyl 4-((1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate, which was directly used in the next step.
[0847] LC-MS: (ESI, m / z): [M+H]+ = 354.3.
[0848] Step 10: preparation of tert-butyl 4-((9-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4- methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate
[0849] DIEA (328 mg, 2.54 mmol) and pentafluorophenyl 3-(2,4-dioxotetrahydropyrimidin-1(2 / / )-yl)-4-methoxybenzoate (601 mg, 1.40 mmol) were added to a solution of tert-butyl 4-((1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate (450 mg, 1.27 mmol) in DMF (5 mL), and the reaction liquid was stirred at room temperature for 2 h. Water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 4-((9-(3-(2,4-dioxotetrahydropyrimidin-1(2 / )-yl)-4-methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate.
[0850] LC-MS: (ESI, m / z): [M+H]+ = 600.3.
[0851] Step 11: preparation of 1-(2-methoxy-5-(3-(piperazin-1-ylmethyl)-1-oxa-9-azaspiro[5.5]undecane- 9-carbonyl)phenyl)dihydropyrimidine-2,4(1 / ,3 / )-dione
[0852] HCl / 1,4-dioxane (1.5 mL) was added to a solution of tert-butyl 4-((9-(3-(2,4- dioxotetrahydropyrimidin-1(2 / )-yl)-4-methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3- yl)methyl)piperazine-1-carboxylate (250 mg, 0.42 mmol) in 1,4-dioxane (3 mL), and the mixture was stirred at 30 °C for 1 h. After the reaction was completed, an aqueous Na2CO3 solution (5 mL) was added to adjust the pH to > 7, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over Na2SO4, filtered, and concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give 1-(2-methoxy-5-(3-(piperazin-1-ylmethyl)-1-oxa-9-azaspiro[5.5]undecane-9-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0853] LC-MS: (ESI, m / z): [M+H]+= 500.3.
[0854] Intermediate 20: preparation of 1-(2-methyl-5-(3-(piperazin-1-ylmethyl)-3Z3-1-oxa-9- azaspiro[5.5]undecane-9-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0855] Step 1: preparation of tert-butyl 4-((1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1- carboxylate 19-P1
[0856] Pd / C (0.20 g) was added to a solution of 19-P1 (1.10 g, 2.50 mmol) in methanol (10 mL), and the mixture was purged with hydrogen gas three times using a hydrogen balloon and stirred at 25 °C for 16 h. The reaction liquid was filtered through celite, and the filtrate was concentrated to give a crude product of tertbutyl 4-((1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate, which was directly used in the next step.
[0857] LC-MS (ESI) m / z: [M+H]+ = 354.3.
[0858] Step 2: preparation of tert-butyl 4-((9-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4- methylbenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate
[0859] DIEA (313 mg, 2.42 mmol) and perfluorophenyl 3-(2,4-dioxotetrahydropyrimidin-1(2 / / )-yl)-4-methylbenzoate (503 mg, 1.21 mmol) were added to a solution of tert-butyl 4-((1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate (430 mg, 1.21 mmol) in DMF (5 mL), and the reaction liquid was stirred at room temperature for 2 h. Water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 4-((9-(3-(2,4-dioxotetrahydropyrimidin-1(2 / )-yl)-4-methylbenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazine-1-carboxylate.
[0860] LC-MS (ESI) m / z: [M+H]+ = 584.3.
[0861] Step 3: preparation of 1-(2-methyl-5-(3-(piperazin-1-ylmethyl)-3Z3-1-oxa-9-azaspiro[5.5]undecane-9-carbonyl)phenyl)dihydropyrimidine-2,4(1 / ,3 / )-dione
[0862] HCl / 1,4-dioxane (5 mL) was added to a solution of tert-butyl 4-((9-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3- yl)methyl)piperazine-1-carboxylate (500 mg, 0.86 mmol) in 1,4-dioxane (5 mL), and the mixture was stirred at 30 °C for 1 h. After the reaction was completed, an aqueous Na2CO3 solution (5 mL) was added to adjust the pH to > 7, and the mixture was extracted with ethyl acetate / isopropanol (20 mL x 3, v / v = 5 / 1). The organic phases were combined, dried over Na2SO4, filtered, and concentrated to give 1-(2-methyl-5-(3-(piperazin-1-ylmethyl)-3Z3-1-oxa-9-azaspiro[5.5]undecane-9-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0863] LC-MS (ESI) m / z: [M+H]+ = 484.3.
[0864] Intermediate 21: (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-3,3-d2-1-ol
[0865] Step 1: preparation of methyl (2S)-2-methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propanoate
[0866] Methyl (5)-3-hydroxy-2-methylpropanoate (3.0 g, 25.39 mmol) was dissolved in DCM (60 mL), and 3,4-dihydropyran (4.27 g, 50.79 mmol) and pyridinium p-toluenesulfonate (1.91 g, 7.62 mmol) were added. The mixture was reacted at room temperature for 2 h. After the reaction was completed, the reaction liquid was poured into an aqueous solution (100 mL), and then the mixture was extracted with dichloromethane (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 100:1 to 5:1) to give methyl (2S)-2-methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propanoate.
[0867] 1H NMR (400 MHz, CDCh) 8 4.68 - 4.55 (m, 1H), 3.95 - 3.73 (m, 2H), 3.69 (s, 3H), 3.63 - 3.41 (m, 2H), 2.85 - 2.70 (m, 1H), 1.82 - 1.48 (m, 6H), 0.65 - 0.95 (m, 3H).
[0868] Step 2: preparation of (2R)-2-methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propan-1,1-d2-1-ol
[0869] Methyl (25)-2-methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propanoate (3.0 g, 14.8 mmol) was dissolved in tetrahydrofuran (80 mL), and the mixture was cooled to 0 °C. LiAlD4 (16 mL, 1.0 M / THF, 16 mmol) was added dropwise, and the mixture was reacted at 0 °C for 15 min. After the reaction was completed, sodium sulfate decahydrate (2 g) was added to quench the reaction, and then the mixture was washed with ethyl acetate (200 mL) and filtered. The filtrate was concentrated to give (2R)-2-methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propan-1,1-d2-1-ol.
[0870] 1H NMR (400 MHz, CDCl3) 8 4.58 (t, J = 3.4 Hz, 1H), 3.92 - 3.62 (m, 2H), 3.60 - 3.30 (m, 2H), 2.11 - 1.93 (m, 1H), 1.90 - 1.68 (m, 2H), 1.62 - 1.50 (m, 4H), 0.93 - 0.78 (m, 3H).
[0871] Step 3: preparation of 1-benzyl-4-((2R)-2-methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propyl-1,1-d2)piperazine
[0872] (2R)-2-Methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propan-1,1-d2-1-ol (2.4 g, 13.62 mmol) was dissolved in dichloromethane (60 mL), and triethylamine (4.13 g, 40.85 mmol) and methanesulfonyl chloride (2.34 g, 20.43 mmol) were added at 0 °C. The mixture was reacted for 1 h. After the reaction was completed, the mixture was warmed to room temperature. Water (100 mL) was added, and the mixture was extracted with ethyl acetate (60 mL x 3). The organic phases were combined, washed with a saturated NaCl solution (100 mL), dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The concentrate was dissolved in DMF (25 mL), and potassium carbonate (5.65 g, 40.85 mmol) and 1-benzylpiperazine (5.79 g, 27.23 mmol) were added. The reaction liquid was heated to 60 °C and reacted for 16 h. After the reaction was completed, the reaction liquid was cooled to room temperature. Water (100 mL) was added, and the mixture was extracted with ethyl acetate (60 mL x 2). The organic phases were combined, washed with a saturated NaCl solution (100 mL), dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give 1-benzyl-4-((2R)-2-methyl-3-((tetrahydro-2H-pyran-2-yl)oxy)propyl-1,1-d2)piperazine.
[0873] LC-MS: (ESI, m / z): [M+H]+ = 335.3.
[0874] Step 4: preparation of (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-3,3-d2-1-ol
[0875] 1-Benzyl-4-((2 R )-2-methyl-3-((tetrahydro-2 H-pyran-2-yl)oxy)propyl-1,1- d2)piperazine (1.5 g, 4.48 mmol) was dissolved in MeOH (14 mL), and concentrated hydrochloric acid (3 mL) was added in an ice bath. The mixture was reacted at room temperature for 1 h. After the reaction was completed, the MeOH and HCl were removed by concentration. An aqueous Na2CO3 solution was added to adjust the pH of the mixture to > 7, and then the mixture was extracted with EtOAc (20 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to give (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-3,3-d2-1-ol.
[0876] LC-MS: (ESI, m / z): [M+H]+ = 251.3.
[0877] Intermediate 22: intermediate 22-P1 and intermediate 22-P2
[0878] Step 1: preparation of tert-butyl 1-(methyl-d3)-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0879] tert-Butyl 3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (11.0 g, 24.2 mmol) was dissolved in tetrahydrofuran (100 mL). TMEDA (5.62 g, 48.4 mmol) was added, and then s-BuLi (27.9 mL, 36.3 mmol, 1.3 M / THF) was added dropwise at -30 °C. After the dropwise addition was completed, the mixture was stirred at 0 °C for 30 min. Deuterated iodomethane (10.52 g, 72.6 mmol) was then added dropwise, and the mixture was stirred at 0 °C for another 15 min. After the reaction was completed, a saturated aqueous ammonium chloride solution (100 mL) was added to the reaction liquid to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EtOAc = 100:1 to 10:1) to give tert-butyl 1-(methyl-d3)-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0880] LC-MS: (ESI, m / z): [M+Na]+ = 494.2.
[0881] Step 2: preparation of tert-butyl 1-(methyl-d3)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc Boc i i
[0882] tert-Butyl 1-(methyl-d3)-3-trityl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (9.0 g, 19.1 mmol) was dissolved in dichloromethane (50 mL), and glacial acetic acid (50 mL) was added. The mixture was refluxed at 50 °C for 1 h. After the reaction was completed, the mixture was concentrated, and the resulting crude product was subjected to silica gel column chromatography (DCM:MeOH (NH3) = 20:1) to give tertbutyl 1-(methyl-d3)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0883] LC-MS: (ESI, m / z): [M+H]+ = 230.2.
[0884] Step 3: preparation of intermediate 22-P1 and intermediate 22-P2 ci 22-P2or22-P1
[0885] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (3.0 g, 11.9 mmol) and triethylamine (3.61 g, 35.7 mmol) were dissolved in tetrahydrofuran (30 mL), and the mixture was cooled to -50 °C. tert-Butyl 1-(methyl- d3)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (2.46 g, 10.71 mmol) was added, and the mixture was reacted at -50 °C for 3 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (50 mL), and then the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was subjected to silica gel column chromatography (PE:EA = 2:1) to give tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methyl-d3)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate. The racemate obtained from the purification was subjected to chiral SFC resolution to give intermediate 22-P1 and intermediate 22-P2. The chiral resolution method was as follows: chromatographic column: Daicel CHIRALCEL IB-N, 250 mm x 30 mm I.D., 10 ^m; mobile phase: CO2 / MeOH [0.2% NH3 (7 M solution in MeOH)]; gradient: 60 / 40, 6 min; flow rate: 100 mL / min; wavelength: 214 nm; column temperature: 35 °C.
[0886] Intermediate 22-P1 (chiral column retention time: 1.260 min). SFC analysis method: column: Daicel CHIRALPAK IB-3, 3 x 150 mm, 3 ^m; mobile phase: CO2 / MeOH; gradient: 70 / 30, 6 min; flow rate: 2 mL / min; wavelength: 214 nm & 254 nm; column temperature: 37 °C; optical purity: 100%.
[0887] Intermediate 22-P2 (chiral column retention time: 1.681 min). SFC analysis method: column: Daicel CHIRALPAK IB-3, 3 x 150 mm, 3 um; mobile phase: CO2 / MeOH; gradient: 70 / 30, 6 min; flow rate: 2 mL / min; wavelength: 214 nm & 254 nm; column temperature: 37 °C; optical purity: 100%.
[0888] LC-MS: (ESI, m / z): [M+H]+ = 445.1.
[0889] Intermediate 23: (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-1-ol
[0890] The intermediate was prepared with reference to the preparation method for intermediate 25 in WO2024083258A1.
[0891] Intermediate 24: 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3- azaspiro[5.5]undecane-9-carbaldehyde-d
[0892] Step 1: preparation of 3-(tert-butoxycarbonyl)-3-azaspiro[5.5]undecane-9-carboxylic acid
[0893] tert-Butyl 9-formyl-3-azaspiro[5.5]undecane-3-carboxylate (2.0 g, 7.11 mmol) was dissolved in tert-butanol (20 mL) and water (10 mL), and 2-methyl-2-butene (1.0 g, 14.22 mmol), sodium chlorite (9.11 g, 21.33 mmol), and sodium dihydrogen phosphate (3.83 g, 31.95 mmol) were added sequentially. After the addition was completed, the mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction liquid was filtered, and the filtrate was concentrated. The resulting crude product was purified by reversed- phase silica gel column chromatography (H2O:ACN = 50:1 to 1:2) to give 3-(tert-butoxycarbonyl)-3-azaspiro[5.5]undecane-9-carboxylic acid.
[0894] LC-MS: (ESI, m / z): [M-H]- = 296.2.
[0895] Step 2: preparation of tert-butyl 9-(hydroxymethyl-d2)-3-azaspiro[5.5]undecane-3-carboxylate
[0896] 3-(tert-Butoxycarbonyl)-3-azaspiro[5.5]undecane-9-carboxylic acid (1.2 g, 4.035 mmol) was dissolved in tetrahydrofuran (12 mL), and LiAlD4 (6 mL, 1.0 M / THF) was slowly added dropwise at 0 °C. The mixture was warmed to room temperature and reacted for 2 h. After the reaction was completed, sodium sulfate decahydrate was added to quench the reaction. The mixture was then filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH = 15:1) to give tert-butyl 9-(hydroxymethyl-d2)-3-azaspiro[5.5]undecane-3-carboxylate.
[0897] LC-MS: (ESI, m / z): [M-56+H]+ = 230.2.
[0898] Step 3: preparation of (3-azaspiro[5.5]undecan-9-yl)methan-d2-ol
[0899] tert-Butyl 9-(hydroxymethyl-d2)-3-azaspiro[5.5]undecane-3-carboxylate (350 mg, 1.23 mmol) was dissolved in DCM (3 mL), and TFA (3 mL) was added. The mixture was reacted at room temperature for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure to give a crude product of (3-azaspiro[5.5]undecan-9-yl)methan-d2-ol. The crude product was directly used in the next step without further purification.
[0900] LC-MS: (ESI, m / z): [M+H]+ = 186.2.
[0901] Step 4: preparation of 1-(5-(9-(hydroxymethyl-d2)-3-azaspiro[5.5]undecane-3-carbonyl)-2- methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0902] The crude product of (3-azaspiro[5.5]undecan-9-yl)methan-d2-ol (350 mg), was dissolved in DMF (5 mL), and then DIEA (793 mg, 6.15 mmol) and pentafluorophenyl 3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoate (582 mg, 1.35 mmol) were added at 0 °C. The reaction liquid was warmed to room temperature and reacted for 2 h. After the reaction was completed, water (40 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated to give a crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give 1-(5-(9-(hydroxymethyl-d2)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0903] LC-MS: (ESI, m / z): [M+H]+ = 432.2.
[0904] Step 5: preparation of 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde-d
[0905] 1-(5-(9-(Hydroxymethyl-d2)-3-azaspiro[5.5]undecane-3-carbonyl)-2- methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione (310 mg, 0.718 mmol) was dissolved in DCM (5 mL), and then the mixture was cooled to 0 °C. Dess-Martin reagent (762 mg, 1.796 mmol) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed, a sodium thiosulfate solution (5 mL) and a sodium bicarbonate solution (5 mL) were added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by reversed-phase column chromatography (H2O:ACN = 20:1 to 1:2) to give 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde-d.
[0906] LC-MS: (ESI, m / z): [M+H]+ = 429.2.
[0907] Intermediate 25: (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-1,1-d2-1-ol D D
[0908] The intermediate was synthesized with reference to the method for intermediate 25 in WO2024083258A1.
[0909] Intermediate 26: intermediate 26-P1 and intermediate 26-P2
[0910] Step 1: preparation of tert-butyl 3-benzyl-1-fluoro-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0911] sec-Butyllithium (6.4 mL, 15.90 mmol, 2.5 M / THF) was added to a solution of tert-butyl 3-benzyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (4 g, 13.25 mmol) and TMEDA (3.07 g, 26.50 mmol) in THF (50 mL) at -78 °C, and the mixture was stirred for 0.5 h. NFSI (6.27 g, 19.88 mmol) was then added at -78 °C, and the mixture was stirred for 1 h. After the reaction was completed, the reaction liquid was poured into a saturated ammonium chloride solution (50 mL), and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (PE:EA = 20:1) to give tert-butyl 3-benzyl-1-fluoro-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0912] LC-MS: (ESI, m / z): [M+H]+ = 321.3.
[0913] Step 2: preparation of tert-butyl 1-fluoro-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0914] Pd / C (2 g) was added to a solution of tert-butyl 3-benzyl-1-fluoro-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (3.9 g, 12.19 mmol) in methanol (50 mL), and the mixture was reacted at 25 °C for 2 h under hydrogen atmosphere. After the reaction was completed, the mixture was filtered and washed with ethyl acetate (10 mL). The filtrate was concentrated under reduced pressure to give tert-butyl 1-fluoro-3,8- diazabicyclo[3.2.1]octane-8-carboxylate.
[0915] LC-MS: (ESI, m / z): [M-56+H]+ = 175.3.
[0916] Step 3: preparation of intermediate 26-P1 and intermediate 26-P2 intermediate 26-P1 or intermediate 26-P2 intermediate 26-P2 or intermediate 26-P1
[0917] 2,4,7-Trichloro-8-fluoropyrido[4,3-d]pyrimidine (2.95 g, 11.74 mmol) was slowly added to a solution of tert-butyl 1-fluoro-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (2.7 g, 11.74 mmol) and triethylamine (2.37 g, 23.48 mmol) in anhydrous tetrahydrofuran (50 mL), and the mixture was stirred at -50 °C for 1 h. A saturated ammonium chloride solution (50 mL) was poured into the reaction liquid, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (PE:EA = 10:1) to give a racemate, tert-butyl 3-(2,7-dichloro-8-fluoropyrido[4,3-^]pyrimidin-4-yl)-1-fluoro-5Z3-3,8-diazabicyclo[3.2.1]octane-8-carboxylate, which was subjected to chiral SFC resolution to give intermediate 26-P1 and intermediate 26-P2. The chiral resolution method was as follows: chromatographic column: Daicel CHIRALCEL OD, 250 mm x 30 mm I.D., 10 ^m; mobile phase: CO2 / MeOH [0.2% NH3 (7 M / MeOH)]; gradient: 80 / 20; flow rate: 150 mL / min; wavelength: 214 nm; column temperature: 35 °C.
[0918] Intermediate 26-P1 (chiral column retention time: 1.693 min). SFC analysis method: column: Daicel Daicel CHIRALCEL OD-3, 150 mm x 3 mm, 3 um; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 60 / 40, 6 min; flow rate: 2 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 100%.
[0919] Intermediate 26-P2 (chiral column retention time: 1.942 min). SFC analysis method: column: Daicel Daicel CHIRALCEL OD-3, 150 mm x 3 mm, 3 um; mobile phase: CO2 / MeOH (0.1% DEA); gradient: 60 / 40, 6 min; flow rate: 2 mL / min; wavelength: 214 nm; column temperature: 37 °C; optical purity: 98.34%.
[0920] LC-MS: (ESI, m / z): [M+H]+ = 446.0.
[0921] Intermediate 27: tert-butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-((R)-2-methyl-3-(4-tritylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9- methanonaphtho[1,8-ab]heptalene-14-carboxylate
[0922] Step 1: preparation of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-(hydroxymethyl)-3,8- diazabicyclo[3.2.1]octane-3,8-dicarboxylate
[0923] tert-Butyl (1S,2 S,5 R )-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.80 g, 3.72 mmol) and sodium bicarbonate (1.87 g, 22.3 mmol) were dissolved in ethyl acetate (20 mL) and water (7 mL), and benzyl chloroformate (1.52 g, 8.92 mmol) was added. The mixture was reacted at room temperature for 18 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (30 mL), and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 2:1) to give 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate.
[0924] LC-MS: (ESI, m / z): [M-Boc+H]+ = 277.1.
[0925] Step 2: preparation of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-formyl-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate
[0926] DMSO (3.89 g, 49.75 mmol) was added dropwise to a solution of oxalyl chloride (1.37 g, 11.72 mmol) in dichloromethane (20 mL) at -78 °C under nitrogen atmosphere. The mixture was stirred at this temperature for 15 min, and then a solution of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-(hydroxymethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate (2.5 g, 6.63 mmol) in DCM (20 mL) was added. The mixture was reacted at -78 °C for 1 h. Triethylamine (5.37 g, 5.31 mmol) was then added, and the mixture was reacted at -78 °C for 2 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous ammonium chloride solution (100 mL), and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 2:1) to give 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-formyl-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate.
[0927] LC-MS: (ESI, m / z): [M+H]+ = 375.2.
[0928] Step 3: preparation of 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8- diazabicyclo[3.2.1]octane-3,8-dicarboxylate
[0929] 3-Benzyl 8-( tert-butyl) (1S,2 S,5 R )-2-formyl-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate (2.03 g, 5.43 mmol) was dissolved in tetrahydrofuran (20 mL), and CH3MgBr (2.17 mL, 6.52 mmol, 3.0 M / THF) was added dropwise at -78 °C. After the dropwise addition was completed, the mixture was stirred at -78 °C for 3 h. After the reaction was completed, a saturated aqueous ammonium chloride solution (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to give a crude product, which was subjected to silica gel column chromatography (PE / EA = 2 / 1) to give 3-benzyl 8-(tert-butyl) (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate.
[0930] LC-MS: (ESI, m / z): [M-56+H]+ = 335.2.
[0931] Step 4: preparation of tert-butyl (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane- 8-carboxylate
[0932] 3-Benzyl 8-( tert-butyl) (1S,2 S,5 R )-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-3,8-dicarboxylate (1.83 g, 4.70 mmol) was dissolved in methanol (20 mL), and then Pd / C (0.2 g) was added. The mixture was purged three times under hydrogen atmosphere and reacted at 25 °C for 16 h. After the reaction was completed, the reaction mixture was filtered and concentrated to give a crude product. The organic phase was concentrated under reduced pressure to give tert-butyl (1S,2S,5R)-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate, which was directly used in the next step without further purification.
[0933] LC-MS: (ESI, m / z): [M+H]+ = 257.2.
[0934] Step 5: preparation of tert-butyl (1S,2S,5R)-2-((S)-1-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0935] tert -Butyl (1S,2 S,5 R )-2-((S)-1-hydroxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.07 g, 4.17 mmol) was dissolved in tetrahydrofuran (20 mL), and the mixture was cooled to 0 °C. NaH (0.5 g, 12.5 mmol, 60%) was added, and the mixture was reacted at 0 °C for 30 min. 5,7-Dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (1.28 g, 4.58 mmol) was then added, and the mixture was reacted at 25 °C for 3 h. After the reaction was completed, water (10 mL) was added to the reaction liquid, and then the tetrahydrofuran was removed from the reaction liquid by distillation under reduced pressure. The mixture was then adjusted to about pH 8 with dilute hydrochloric acid (1 M) until a precipitate was formed. The precipitate was collected by filtration and dried to give a crude product of tert-butyl (1S,2S,5R)-2-((S)-1-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate. The crude product was directly used in the next step without further purification.
[0936] LC-MS: (ESI, m / z): [M+H]+ = 500.1.
[0937] Step 6: preparation of tert-butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate F
[0938] tert -Butyl (1S,2 S,5 R )-2-((S)-1-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3- ^]pyrimidin-5-yl)oxy)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.43 g, 2.87 mmol) and DIEA (0.37 g, 28.7 mmol) were dissolved in DCM (20 mL), and then BOPCl (2.19 g, 8.6 mmol) was added to the mixture. The mixture was reacted at 25 °C for 4 h. After the reaction was completed, water (50 mL) was added to the reaction liquid, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate.
[0939] LC-MS: (ESI, m / z): [M+H]+ = 482.1.
[0940] Step 7: preparation of tert-butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate Boc Boc ! '
[0941] tert-Butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylthio)-5a,6,7,8,9,10-hexahydro- 5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate (0.87 g, 1.8 mmol) was dissolved in dichloromethane (10 mL), and m-CPBA (0.47 g, 2.7 mmol) was added. The mixture was reacted at room temperature for 2 h. After the reaction was completed, the reaction liquid was poured into a saturated aqueous sodium thiosulfate solution (20 mL), and the mixture was washed with a saturated aqueous sodium bicarbonate solution (10 mL) and then extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated to give a crude product of tert-butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate . The crude product was directly used in the next step without purification.
[0942] LC-MS: (ESI, m / z): [M+H]+ = 498.1.
[0943] Step 8: preparation of tert-butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-((R)-2-methyl-3-(4-tritylpiperazin-1-yl)propoxy)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate
[0944] (R )-2-Methyl-3-(4-tritylpiperazin-1-yl)propan-1-ol (0.8 g, 2.0 mmol) was dissolved in tetrahydrofuran (20 mL), and the mixture was cooled to 0 °C. NaH (132 mg, 3.3 mmol) was slowly added, and the mixture was reacted at 25 °C for 30 min. tert-Butyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-(methylsulfinyl)-5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate (0.83 g, 1.67 mmol) was then added, and the mixture was reacted at 0 °C for 1 h. After the reaction was completed, a saturated aqueous ammonium chloride solution (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH = 15:1) to give tertbutyl (5S,5aS,6S,9R)-2-chloro-1-fluoro-5-methyl-12-((R)-2-methyl-3-(4-tritylpiperazin-1-yl)propoxy)- 5a,6,7,8,9,10-hexahydro-5H-4-oxa-3,10a,11,13,14-pentaaza-6,9-methanonaphtho[1,8-ab]heptalene-14-carboxylate.
[0945] LC-MS: (ESI, m / z): [M+H]+ = 834.3.
[0946] Example 1: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0947] Step 1: preparation of tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4- methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0948] Tetraisopropyl titanate (0.5 mL) was added to a stirred solution of intermediate 12 (200 mg, 0.21 mmol) and 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde (100 mg, 0.23 mmol) in DCM / AcOH (2 mL / 0.2 mL), and the mixture was stirred at 30 °C for 2 h. NaBH(OAc)3 (135 mg, 0.64 mmol) was added to the mixture in an ice bath, and the resulting mixture was stirred for 2 h. The mixture was concentrated, and the resulting concentrate was purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0949] LC-MS: (ESI, m / z): [M / 2+H]+ = 678.5.
[0950] Step 2: preparation of 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0951] Cesium fluoride (336 mg, 2.2 mmol) was added to a stirred solution of tert -butyl 3-(2-((R )-3-(4-((3- (3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9- yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (200 mg, 0.15 mmol) in DMF (2 mL), and the mixture was stirred at room temperature for 1 h. Water (10 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. HCl / dioxane (6 N, 1.0 mL) was added to a stirred solution of the concentrate in 1,4-dioxane (1.0 mL), and the mixture was reacted at room temperature for 0.5 h. An aqueous Na2CO3 solution was added to the reaction liquid to adjust the pH to > 7, and the mixture was extracted with EtOAc (10 mL x 3). The organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by preparative high-performance liquid chromatography to give 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0952] LC-MS: (ESI, m / z): [M / 2+H]+ = 528.6.
[0953] 1H NMR (400 MHz, DMSO-d6) 5 10.33 (s, 1H), 10.15 (s, 1H), 9.02 (s, 1H), 7.98 (dd, J = 9.2, 6.0 Hz, 1H), 7.46 (t, J = 9.0 Hz, 1H), 7.41 - 7.30 (m, 3H), 7.21 - 7.11 (m, 2H), 4.59 - 4.26 (m, 3H), 4.13 - 4.01 (m, 1H), 3.91 (d, J = 29.6 Hz, 1H), 3.84 (s, 3H), 3.69 - 3.52 (m, 5H), 3.51 - 3.35 (m, 6H), 3.33 (s, 4H), 2.71 - 2.65 (m, 2H), 2.46 - 1.98 (m, 12H), 1.78 - 1.62 (m, 5H), 1.56 - 1.38 (m, 6H), 1.36 - 1.16 (m, 3H), 1.11 - 1.00 (m, 4H), 0.97 (d, J = 6.0 Hz, 3H).
[0954] Example 2: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0955] Step 1: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc i HO Bn intermediate 2-P2
[0956] Intermediate 2-P2 (400 mg, 0.85 mmol) and (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-1-ol (421 mg, 1.69 mmol) were dissolved in THF (5 mL), and the mixture was added to a sealed tube reactor. Cs2CO3 (828 mg, 2.54 mmol) was then added, and the mixture was reacted at 90 °C for 16 h. The reaction liquid was cooled to room temperature, and water (10 mL) was added. The mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EtOAc = 1:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0957] LC-MS: (ESI, m / z): [M+H]+ = 684.3.
[0958] Step 2: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7- (7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate TIPS Bn OMOM
[0959] CataCXium A Pd-G3 (85 mg, 0.12 mmol) and cesium carbonate (571 mg, 1.75 mmol) were added to a mixed solution of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (395 mg, 0.58 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (360 mg, 0.7 mmol) in 1,4-dioxane (3 mL) and water (0.6 mL). The mixture was reacted at 85 °C for 16 h under nitrogen atmosphere. Water (10 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0960]
[0961] LC-MS: (ESI, m / z): [M / 2+H]+ = 517.8. Step 3: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (736 mg, 0.71 mmol) was dissolved in dichloromethane (35 mL), and then 1-chloroethyl chloroformate (206 mg, 1.44 mmol) and N,N-diisopropylethylamine (272 mg, 2.10 mmol) were added. The mixture was reacted at room temperature for 1 h and concentrated under reduced pressure. The concentrate was dissolved in methanol (35 mL), and the resulting mixture was heated to 50 °C and reacted for 30 min. After the reaction was completed, the mixture was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0963] LC-MS: (ESI, m / z): [M+H]+ = 944.4.
[0964] Step 4: preparation of tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0965] Tetraisopropyl titanate (226 mg, 0.79 mmol) was added to a mixed solution of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (150 mg, 0.16 mmol) and 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde (102 mg, 0.24 mmol) in dichloromethane (2 mL) and glacial acetic acid (0.2 mL), and the mixture was stirred at 30 °C for 2 h. Sodium triacetoxyborohydride (67 mg, 0.32 mmol) was added to the mixture in an ice bath, and the resulting mixture was stirred for 2 h. The mixture was concentrated, and the resulting crude product was purified by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to give tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0966] LC-MS: (ESI, m / z): [M / 2+H]+ = 678.5.
[0967] Step 5: preparation of 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0968] Cesium fluoride (234 mg, 1.54 mmol) was added to a stirred solution of tert-butyl 3-(2-((R )-3-(4- ((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9- yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (208 mg, 0.15 mmol) in N,N-dimethylformamide (5 mL), and the mixture was stirred at room temperature for 1 h. Water (30 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. Hydrogen chloride / 1,4-dioxane (6 N, 1.0 mL) was added to a stirred solution of the concentrate in 1,4-dioxane (1.0 mL), and the mixture was reacted at room temperature for 0.5 h. An aqueous Na2CO3 solution was added to the reaction liquid to adjust the pH to > 7, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by preparative high-performance liquid chromatography to give 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0969] LC-MS: (ESI, m / z): [M / 2+H]+ = 528.3.
[0970] 1H NMR (400 MHz, DMSO-d6) 8 10.33 (s, 1H), 10.15 (s, 1H), 9.02 (s, 1H), 7.98 (dd, J = 9.2, 6.0 Hz, 1H), 7.46 (t, J = 9.2 Hz, 1H), 7.39 (d, J = 2.4 Hz, 1H), 7.36 (dd, J = 8.4, 2.0 Hz, 1H), 7.31 (d, J = 2.0 Hz, 1H), 7.20 - 7.17 (m, 1H), 7.14 (d, J = 8.8 Hz, 2H), 4.54 - 4.32 (m, 3H), 4.08 - 4.02 (m, 1H), 3.94 - 3.87 (m, 1H), 3.84 (s, 3H), 3.64 - 3.55 (m, 5H), 3.55 - 3.41 (m, 6H), 3.40 - 3.34 (m, 4H), 2.69 - 2.66 (m, 2H), 2.32 -2.05 (m, 12H), 1.75 - 1.65 (m, 5H), 1.54 - 1.38 (m, 6H), 1.38 - 1.19 (m, 3H), 1.15 - 0.97 (m, 4H), 0.97 (d, J = 6.0 Hz, 3H).
[0971] Example 18: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0972] Step 1: preparation of tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0973] Tetraisopropyl titanate (301 mg, 1.05 mmol) was added to a solution of intermediate 12 (200 mg, 0.21 mmol) and 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecane-9- carbaldehyde (105 mg, 0.25 mmol) in DCM / AcOH (1.5 mL / 0.15 mL), and the mixed solution was stirred at 25 °C for 2 h. NaBH(OAc)3 (135 mg, 0.64 mmol) was then added at 0 °C, and the mixture was stirred for 1 h. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3- azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0974] LC-MS: (ESI, m / z): [M / 2+H]+ = 670.5.
[0975] Step 2: preparation of 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0976] Cesium fluoride (289 mg, 1.90 mmol) was added to a solution of tert -butyl 3-(2-((R )-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (170 mg, 0.13 mmol) in DMF (2 mL), and the mixture was stirred at 35 °C for 1 h. The reaction liquid was diluted with H2O (5 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. 1,4-Dioxane (1 mL) and hydrochloric acid / 1,4-dioxane (4 M, 1 mL) were added to the crude product, and the mixture was stirred at room temperature for 0.5 h. The reaction liquid was adjusted to pH > 7 with an aqueous Na2CO3 solution, and the mixture was extracted with ethyl acetate (15 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by preparative high-performance liquid chromatography to give 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3- hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[0977] 1H NMR (400 MHz, DMSO-d6) 5 10.38 (s, 1H), 10.16 (s, 1H), 9.02 (s, 1H), 7.98 (dd, J = 9.1, 6.0 Hz, 1H), 7.52 - 7.42 (m, 1H), 7.39 (d, J = 2.4 Hz, 1H), 7.36 - 7.28 (m, 2H), 7.26 - 7.16 (m, 2H), 4.60 - 4.28 (m, 3H), 4.12 - 4.01 (m, 1H), 3.91 (d, J = 30.0 Hz, 1H), 3.85 - 3.74 (m, 1H), 3.69 - 3.36 (m, 10H), 3.18 - 3.30 (m, 2H), 2.83 - 2.63 (m, 3H), 2.47 - 2.22 (m, 9H), 2.21 (s, 3H), 2.20 - 1.95 (m, 5H), 1.77 - 1.62 (m, 5H), 1.56 - 1.38 (m, 6H), 1.37 - 1.17 (m, 3H), 1.13 - 1.00 (m, 3H), 0.97 (d, J = 6.0 Hz, 3H).
[0978] LC-MS: (ESI, m / z): [M+H]+ = 1039.90.
[0979] Example 19: compound 19-P1 and compound 19-P2
[0980] Step 1: preparation of tert-butyl 3-(2-((2R)-3-(4-((9-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[0981] Tetraisopropyl titanate (619 mg, 2.18 mmol) was added to a solution of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1- yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (280 mg, 0.31 mmol) and 9-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecane-3-carbaldehyde (107 mg, 0.26 mmol) in DCM / AcOH (3 mL / 0.3 mL), and the mixture was stirred at 25 °C for 2 h. NaBH(OAc)3 (110 mg, 0.52 mmol) was added to the mixture in an ice bath, and the resulting mixture was stirred for 2 h. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 3-(2-((2R)-3-(4-((9-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[0982] LC-MS: (ESI, m / z): [M / 2+H]+ = 657.5.
[0983] Step 2: preparation of compound 19-P1 and compound 19-P2
[0984] Cesium fluoride (121 mg, 0.80 mmol) was added to a solution of tert-butyl 3-(2-((2R)-3-(4-((9-(3- (2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-1-oxa-9-azaspiro[5.5]undecan-3- yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (210 mg, 0.16 mmol) in DMF (3 mL), and the mixture was stirred at room temperature for 1 h. Water (10 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. HCl / 1,4-dioxane (6 N, 2.0 mL) was added to a stirred solution of the concentrate in 1,4-dioxane (2.0 mL), and the mixture was reacted at room temperature for 0.5 h. An aqueous Na2CO3 solution was added to the reaction liquid to adjust the pH to > 7, and the mixture was extracted with EtOAc (20 mL x 3). The organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by preparative high-performance liquid chromatography to give a racemate, 1-(5-(3-((4-((R)-3-((4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-1-oxa-9- azaspiro[5.5]undecane-9-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione. The racemic compound obtained from the purification was subjected to SFC resolution with the following method: chromatographic column: CHIRALPAK IC (IC00CD-TB016), 0.46 cm x 15 cm I.D., 10 ^m; mobile phase: MeOH / ACN / DEA; gradient: 65 / 35 / 0.2 (V / V / V); flow rate: 1 mL / min; wavelength: 254 nm; column temperature: 35 °C.
[0985] Compound 19-P1 (chiral column retention time: 6.913 min). Chiral analysis method: chromatographic column: CHIRALPAK IC(IC00CD-TB016), 0.46 cm x 15 cm I.D., 10 ^m; mobile phase: MeOH / ACN / DEA; gradient: 65 / 35 / 0.2 (V / V / V); flow rate: 1 mL / min; wavelength: 254 nm; column temperature: 35 °C; optical purity: 93%.
[0986] LC-MS: (ESI, m / z): [M+H]+ = 1013.5.
[0987] 1H NMR (400 MHz, CD3OD) 5 9.01 (s, 1H), 7.86 (dd, J = 9.1, 5.7 Hz, 1H), 7.44 (dd, J = 8.5, 2.0 Hz, 1H), 7.39 (d, J = 2.0 Hz, 1H), 7.37 - 7.29 (m, 2H), 7.24 - 7.13 (m, 2H), 4.71 - 4.49 (m, 4H), 4.39 - 4.13 (m, 2H), 3.92 (s, 3H), 3.84 - 3.55 (m, 8H), 3.37 (s, 1H), 3.35 - 3.31 (m, 2H), 2.80 (t, J = 6.7 Hz, 2H), 2.75 -2.40 (m, 9H), 2.40 - 2.08 (m, 5H), 1.97 - 1.50 (m, 9H), 1.50 - 1.28 (m, 3H), 1.09 (d, J = 6.0 Hz, 3H).
[0988] Compound 19-P2 (chiral column retention time: 9.062 min). Chiral analysis method: chromatographic column: CHIRALPAK IC(IC00CD-TB016), 0.46 cm x 15 cm I.D., 10 um; mobile phase: MeOH / ACN / DEA; gradient: 65 / 35 / 0.2 (V / V / V); flow rate: 1 mL / min; wavelength: 254 nm; column temperature: 35 °C; optical purity: 97%.
[0989] LC-MS: (ESI, m / z): [M+H]+ = 1013.5.
[0990] 1H NMR (400 MHz, CD3OD) 5 9.00 (s, 1H), 7.86 (dd, J = 9.1, 5.7 Hz, 1H), 7.44 (dd, J = 8.5, 2.0 Hz, 1H), 7.39 (d, J = 2.0 Hz, 1H), 7.37 - 7.29 (m, 2H), 7.24 - 7.13 (m, 2H), 4.68 - 4.60 (m, 2H), 4.58 - 4.50 (m, 2H), 4.39 - 4.13 (m, 2H), 3.92 (s, 3H), 3.84 - 3.55 (m, 8H), 3.37 (s, 1H), 3.35 - 3.31 (m, 2H), 2.80 (t, J = 6.7 Hz, 2H), 2.75 - 2.40 (m, 9H), 2.40 - 2.08 (m, 5H), 1.97- 1.50 (m, 9H), 1.50 - 1.28 (m, 3H), 1.09 (d, J = 6.0 Hz, 3H).
[0991] Example 49: 1-(5-(9-((4-((S)-3-((4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethynyl-7-fluoro-3- hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methoxypropyl)piperazin-1- yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[0992] Step 1: preparation of (S)-1-(4-benzylpiperazin-1-yl)-3-((tert-butyldimethylsilyl)oxy)propan-2-ol
[0993] A mixed solution of (5)-tert-butyldimethyl(oxiran-2-ylmethoxy)silane (20 g, 106 mmol), 1-benzylpiperazine hydrochloride (25 g, 117 mmol), and potassium carbonate (29 g, 212 mmol) in THF (15 mL) was heated to reflux for 3 h. After the reaction was completed, the mixed solution was filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EtOAc = 2:1) to give (5)-1-(4-benzylpiperazin-1-yl)-3-((tert-butyldimethylsilyl)oxy)propan-2-ol.
[0994] LC-MS: (ESI, m / z): [M+H]+ = 365.3.
[0995] Step 2: preparation of (5)-1-benzyl-4-(3-((tert-butyldimethylsilyl)oxy)-2-methoxypropyl)piperazine TBSO^^N^i ______ TBSO^^^N^ °H ^'Bn " °" ^'Bn
[0996] (5)-1-(4-Benzylpiperazin-1-yl)-3-(( tert-butyldimethylsilyl)oxy)propan-2-ol (5.00 g, 13.70 mmol) was dissolved in THF (50 mL), and NaH (0.82 g, 20.55 mmol) was added at 0 °C. The mixture was reacted at 25 °C for 30 min and then cooled to 0 °C. Iodomethane (2.14 g, 15.07 mmol) was added, and the mixture was reacted at room temperature for 3 h. After the reaction was completed, the reaction liquid was slowly poured into a cooled aqueous NH4Cl solution (100 mL), and then the mixture was extracted with ethyl acetate (50 mL x 2). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 30:1 to 5:1) to give (5)-1-benzyl-4-(3-((tert-butyldimethylsilyl)oxy)-2-methoxypropyl)piperazine.
[0997] 1H NMR (400 MHz, CDCh) 5 7.32 - 7.28 (m, 4H), 7.26 - 7.22 (m, 1H), 3.63 (d, J = 5.2 Hz, 2H), 3.50 (s, 2H), 3.42 (s, 3H), 3.35 - 3.39 (m, 1H), 2.42 - 2.60 (m, 8H), 0.89 (m, 9H), 0.05 (s, 6H).
[0998] Step 3: preparation of (5)-3-(4-benzylpiperazin-1-yl)-2-methoxypropan-1-ol TBSO^^N^i o. <ABn ----- ^Bn
[0999] (5)-1-Benzyl-4-(3-(( tert-butyldimethylsilyl)oxy)-2-methoxypropyl)piperazine (3.20 g, 8.40 mmol) was dissolved in ethanol (30 mL), and the mixture was cooled to 0-5 °C. Concentrated hydrochloric acid (5 mL) was added, and the mixture was reacted at room temperature for 1 h. The reaction liquid was concentrated to remove the ethanol, and the residue was dissolved in water (100 mL). The mixture was then extracted with methyl tert-butyl ether (50 mL). The aqueous phase was adjusted to pH 8-9 with an aqueous sodium carbonate solution, and the mixture was extracted with ethyl acetate (100 mL X 3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to give (S)-3-(4-benzylpiperazin-1-yl)-2-methoxypropan-1-ol. The resulting product was directly used in the next step without further purification.
[01000] Step 4: preparation of tert-butyl 3-(2-((S)-3-(4-benzylpiperazin-1-yl)-2-methoxypropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01001] tert-Butyl 3-(2,7-dichloro-8-fluoropyrido[4,3- ^]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.70 g, 1.63 mmol) and (5)-3-(4-benzylpiperazin-1-yl)-2-methoxypropan-1-ol (0.65 g, 2.45 mmol) were dissolved in THF (15 mL), and Cs2CO3 (1.06 g, 3.26 mmol) was added. The mixture was reacted at 85 °C for 16 h. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl 3-(2-((S)-3-(4-benzylpiperazin-1-yl)-2-methoxypropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01002] LC-MS: (ESI, m / z): [M+H]+ = 656.3.
[01003] Step 5: preparation of tert-butyl 3-(2-((S)-3-(4-benzylpiperazin-1-yl)-2-methoxypropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01004] CataCXium Pd G3 (55 mg, 0.076 mmol) and cesium carbonate (0.49 g, 1.52 mmol) were added to a mixed solution of tert-butyl 3-(2-((S)-3-(4-benzylpiperazin-1-yl)-2-methoxypropoxy)-7-chloro-8- fluoropyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.50 g, 0.76 mmol) and ((2- fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1- yl)ethynyl)triisopropylsilane (0.51 g, 0.99 mmol) in 1,4-dioxane (6 mL) and H2O (0.8 mL). The mixture was reacted at 85 °C for 16 h under nitrogen atmosphere. Water (20 mL) was added to the reaction liquid, and then the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(2-((S)-3-(4-benzylpiperazin-1-yl)-2-methoxypropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1- yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01005] LC-MS: (ESI, m / z): [M+H]+ = 1006.6.
[01006] Step 6: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((S)-2-methoxy-3-(piperazin-1-yl)propoxy)pyrido[4,3- d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01007] tert-Butyl 3-(2-((S)-3-(4-benzylpiperazin-1-yl)-2-methoxypropoxy)-8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.30 g, 0.30 mmol) was dissolved in DCM (2 mL), and 1-chloroethyl carbonochloridate (64 mg, 0.45 mmol) and DIEA (77 mg, 0.60 mmol) were added. The mixture was reacted at room temperature for 1 h and concentrated under reduced pressure. MeOH (2 mL) was added to the concentrate for dissolution, and then the mixture was heated to 50 °C and reacted for 30 min. After the reaction was completed, the mixture was directly concentrated under reduced pressure to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((S)-2-methoxy-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01008] LC-MS: (ESI, m / z): [M+H]+ = 916.5.
[01009] Step 7: preparation of tert-butyl 3-(2-((S)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methoxypropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01010] Tetraisopropyl titanate (0.5 mL) was added to a solution of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((S)-2-methoxy-3-(piperazin-1- yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (120 mg, 0.13 mmol) and 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde (81 mg, 0.20 mmol) in DCM / AcOH (1.5 mL / 0.1 mL), and the mixture was stirred at 30 °C for 2 h. NaBH(OAc)3 (55 mg, 0.26 mmol) was added to the mixture in an ice bath, and the resulting mixture was stirred for 2 h. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 10:1) to give tert-butyl 3- (2-((S)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9- yl)methyl)piperazin-1-yl)-2-methoxypropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01011] LC-MS: (ESI, m / z): [M / 2+H]+ = 656.5.
[01012] Step 8: preparation of 1-(5-(9-((4-((S)-3-((4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methoxypropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01013] Cesium fluoride (55 mg, 0.36 mmol) was added to a solution of tert-butyl 3-(2-(( 5)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methoxypropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (95 mg, 0.072 mmol) in DMF (1 mL), and the mixture was stirred at room temperature for 30 min. Water (10 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. HCl / 1,4-dioxane (4 N, 1.0 mL) was added to a solution of the concentrate in 1,4-dioxane (2 mL), and the mixture was reacted at room temperature for 0.5 h. The reaction liquid was adjusted to pH > 7 with an aqueous Na2CO3 solution, and the mixture was extracted with EtOAc (20 mL x 2). The organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by preparative high-performance liquid chromatography to give 1-(5-(9-((4-((5)-3-((4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoropyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methoxypropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[01014] LC-MS: (ESI, m / z): [M / 2+H]+ = 506.3.
[01015] 1H NMR (400 MHz, DMSO-d6) 5 10.37 (s, 1H), 10.17 (s, 1H), 9.05 (s, 1H), 7.97 (dd, J = 9.2, 6.0 Hz, 1H), 7.46 (t, J = 8.8 Hz, 1H), 7.39 (d, J = 2.4 Hz, 1H), 7.36 - 7.29 (m, 2H), 7.24 (d, J = 7.6 Hz, 1H), 7.18 (d, J = 2.4 Hz, 1H), 4.62 - 4.44 (m, 2H), 4.39 - 4.25 (m, 2H), 3.93 (s, 1H), 3.83 - 3.75 (m, 1H), 3.73 - 3.43 (m, 10H), 3.36 (s, 3H), 2.88 - 2.62 (m, 3H), 2.47 - 2.24 (m, 10H), 2.21 (s, 3H), 2.11 - 1.97 (m, 2H), 1.79 - 1.58 (m, 6H), 1.57 - 1.37 (m, 5H), 1.36 - 1.19 (m, 2H), 1.16 - 0.87 (m, 4H).
[01016] Example 50: 1-(5-(9-((4-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1- (methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01017] Step 1: preparation of tert-butyl 3-(2-((1-((4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)methyl)cyclopropyl)methoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01018] Tetraisopropyl titanate (320 mg, 1.13 mmol) was added to a stirred solution of intermediate 3 (200 mg, 0.23 mmol) and 1-(2-methoxy-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (202 mg, 0.41 mmol) in dichloromethane / acetic acid (10 mL / 0.1 mL), and the mixture was stirred at 25 °C for 2 h. Sodium triacetoxyborohydride (143 mg, 0.67 mmol) was added to the mixture in an ice bath, and the resulting mixture was stirred for 16 h. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 3-(2-((1-((4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)methyl)cyclopropyl)methoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01019] LC-MS: (ESI, m / z): [M / 2+H]+ = 684.5.
[01020] Step 2: preparation of 1-(5-(9-((4-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01021] Cesium fluoride (50 mg, 0.33 mmol) was added to a solution of tert -butyl 3-(2-((1-((4-((3-(3-(2,4- dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)methyl)cyclopropyl)methoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (45 mg, 0.03 mmol) in N,N-dimethylformamide (5 mL), and the mixture was stirred at room temperature for 2 h. Water (50 mL x 2) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. HCl / 1,4- dioxane (4.0 M, 2.0 mL) was added to a solution of the concentrate in 1,4-dioxane (2.0 mL) in an ice-water bath, and then the mixture was reacted at room temperature for 2 h. The reaction liquid was concentrated under reduced pressure at 30 °C, then diluted with a solution of ammonia in methanol (2.0 M, 5 mL), and concentrated to dryness again. The crude product was purified by preparative high-performance liquid chromatography to give 1-(5-(9-((4-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[01022] LC-MS: (ESI, m / z): [M+H]+ = 1067.8.
[01023] 1H NMR (400 MHz, DMSO-d6) 5 8.98 (d, J = 4.2 Hz, 1H), 7.85 (dd, J = 9.2, 6.0 Hz, 1H), 7.43 (dd, J = 8.4, 2.0 Hz, 1H), 7.38 - 7.29 (m, 3H), 7.21 (d, J = 2.4 Hz, 1H), 7.18 (d, J = 8.8 Hz, 1H), 4.68 - 4.53 (m, 4H), 4.38 - 4.35 (m, 1H), 3.92 (s, 3H), 3.72 - 3.53 (m, 9H), 3.47 - 3.33 (m, 6H), 2.82 - 2.78 (m, 2H), 2.55 -2.35 (m, 7H), 2.22 - 2.13 (m, 2H), 2.05 - 1.85 (m, 2H), 1.78 - 1.41 (m, 13H), 1.25 - 1.08 (m, 4H), 0.75 - 0.65 (m, 2H), 0.55 - 0.45 (m, 2H).
[01024] Example 63: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1- methyl-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1- yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01025] Step 1: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro- 8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc Boc Bn intermediate 14-P1
[01026] Intermediate 14-P1 (500 mg, 1.13 mmol) and (R)-3-(4-benzylpiperazin-1-yl)-2-methylpropan-1-ol (562 mg, 2.26 mmol) were dissolved in THF (5 mL), and the mixture was added to a sealed tube reactor. Cs2CO3 (1102 mg, 3.38 mmol) was then added. The mixture was reacted at 90 °C for 16 h. After the reaction was completed, the mixture was cooled to room temperature. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01027] LC-MS: (ESI, m / z): [M+H]+ = 654.3.
[01028] Step 2: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7- (7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)- 1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01029] CataCXium A Pd G3 (111 mg, 0.15 mmol) and cesium carbonate (745 mg, 2.29 mmol) were added to a mixed solution of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8- fluoropyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (500 mg, 0.76 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (469 mg, 0.91 mmol) in 1,4-dioxane (3 mL) and H2O (0.6 mL). The mixture was reacted at 85 °C for 16 h under nitrogen atmosphere. Water (10 mL) was added, and then the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01030] LC-MS: (ESI, m / z): [M+H]+ = 1004.4.
[01031] Step 3: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate Boc Boc OMOM OMOM
[01032] tert-Butyl 3 -(2-((R )-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (550 g, 0.55 mmol) was dissolved in DCM (5 mL), and 1-chloroethyl chloroformate (234 mg, 1.64 mmol) and DIEA (212 mg, 1.64 mmol) were added. The mixture was reacted at room temperature for 1 h and concentrated under reduced pressure. MeOH (5 mL) was added to the concentrate for dissolution, and the mixture was heated to 50 °C and reacted for 10 min. After the reaction was completed, the mixture was concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3- d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01033] LC-MS: (ESI, m / z): [M+H]+ = 914.4.
[01034] Step 4: preparation of tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01035] Tetraisopropyl titanate (0.5 mL) was added to a stirred solution of tert -butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (150 mg, 0.16 mmol) and 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecane-9-carbaldehyde (84 mg, 0.20 mmol) in DCM / AcOH (2 mL / 0.2 mL), and the mixture was stirred at 30 °C for 2 h. NaBH(OAc)3 (104 mg, 0.49 mmol) was added to the mixture in an ice bath, and the resulting mixture was stirred for 2 h. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01036] LC-MS: (ESI, m / z): [M / 2+H]+ = 663.5.
[01037] Step 5: preparation of 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-methyl-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01038] Cesium fluoride (292 mg, 1.92 mmol) was added to a stirred solution of tert-butyl 3-(2-((R )-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-methyl-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (170 mg, 0.13 mmol) in DMF (2 mL), and the mixture was stirred at room temperature for 1 h. Water (10 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. HCl / 1,4-dioxane (6 N, 1.0 mL) was added to a stirred solution of the concentrate in 1,4-dioxane (1.0 mL), and the mixture was reacted at room temperature for 0.5 h. An aqueous Na2CO3 solution was added to the reaction liquid to adjust the pH to > 7, and the mixture was extracted with EtOAc (10 mL x 3). The organic phase was washed with brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by preparative high-performance liquid chromatography to give 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-methyl-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2-methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[01039] LC-MS: (ESI, m / z): [M / 2+H]+ = 513.3.
[01040] 1H NMR (400 MHz, DMSO-d6) 5 10.33 (s, 1H), 9.04 (d, J = 2.4 Hz, 1H), 7.96 (dd, J = 9.1, 6.0 Hz, 1H), 7.45 (t, J = 9.0 Hz, 1H), 7.39 - 7.33 (m, 2H), 7.32 (d, J = 1.8 Hz, 1H), 7.20 - 7.10 (m, 2H), 4.53 - 4.38 (m, 2H), 4.35 - 4.25 (m, 1H), 4.12 - 4.00 (m, 1H), 3.92 (s, 1H), 3.84 (s, 3H), 3.67 - 3.48 (m, 6H), 3.30 - 3.17 (m, 3H), 2.70 - 2.64 (m, 2H), 2.49 - 2.09 (m, 12H), 2.09 - 2.00 (m, 2H), 1.83 - 1.60 (m, 5H), 1.57 - 1.39 (m, 5H), 1.38 - 1.27 (m, 3H), 1.24 (s, 3H), 1.11 - 0.92 (m, 7H).
[01041] Example 72: 1-(5-(9-((4-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2- yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01042] Step 1: preparation of tert-butyl 3-(2-((1-((4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)methyl)cyclopropyl)methoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01043] Tetraisopropyl titanate (320 mg, 1.13 mmol) was added to a solution of intermediate 3 (200 mg, 0.23 mmol) and 1-(2-methyl-5-(9-(piperazin-1-ylmethyl)-3-azaspiro[5.5]undecane-3- carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (217 mg, 0.45 mmol) in dichloromethane / acetic acid (10 mL / 0.1 mL), and the mixture was stirred at 25 °C for 2 h. Sodium triacetoxyborohydride (143 mg, 0.67 mmol) was added to the mixture in an ice bath, and the resulting mixture was stirred for 16 h. The mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 100:1 to 10:1) to give tert-butyl 3-(2-((1-((4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)methyl)cyclopropyl)methoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01044] LC-MS: (ESI, m / z): [M / 2+H]+ = 676.5.
[01045] Step 2: preparation of 1-(5-(9-((4-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01046] Cesium fluoride (133 mg, 0.88 mmol) was added to a solution of tert -butyl 3-(2-((1-((4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)methyl)cyclopropyl)methoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (118 mg, 0.03 mmol) in N,N-dimethylformamide (5 mL), and the mixture was stirred at room temperature for 2 h. Water (2 x 50 mL) was added to the reaction liquid, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated. HCl / 1,4-dioxane (4.0 M, 2.0 mL) was added to a solution of the concentrate in 1,4-dioxane (2.0 mL) in an ice-water bath, and then the mixture was reacted at room temperature for 2 h. The reaction liquid was concentrated under reduced pressure at 30 °C, then diluted with a solution of ammonia in methanol (2.0 M, 5 mL), and concentrated to dryness again. The crude product was purified by preparative high-performance liquid chromatography to give 1-(5-(9-((4-((1-(((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1-(methoxymethyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-methylphenyl)dihydropyrimidine-2,4(1H,3H)-dione.
[01047] LC-MS: (ESI, m / z): [M+H]+ = 1051.8.
[01048] 1H NMR (400 MHz, DMSO-d6) 5 8.98 (d, J = 4.2 Hz, 1H), 7.85 (dd, J = 9.2, 6.0 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.33 - 7.29 (m, 4H), 7.21 (d, J = 2.4 Hz, 1H), 4.71 - 4.45 (m, 4H), 4.38 - 4.35 (m, 1H), 3.87 -3.83 (m, 1H), 3.72 - 3.47 (m, 9H), 3.41 - 3.33 (m, 6H), 2.88 - 2.81 (m, 2H), 2.54 - 2.36 (m, 7H), 2.31 (s, 3H), 2.15 - 2.09 (m, 2H), 1.95 - 1.80 (m, 2H), 1.75 - 1.29 (m, 12H), 1.25 - 1.05 (m, 4H), 0.75 - 0.65 (m, 2H), 0.55 - 0.45 (m, 2H).
[01049] Example 88: 1-(5-(9-((4-((2R)-3-((7-(8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl)-8-fluoro-4-(1- ((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)pyrido[4,3-d]pyrimidin-2-yl)oxy)-2- methylpropyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)-2- methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione
[01050] Step 1: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro- 8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01051] Boc Intermediate 11-P1 (200 mg, 0.42 mmol) and (R )-3-(4-benzylpiperazin-1-yl)-2-methylpropan-1-ol (209 mg, 0.84 mmol) were dissolved in THF (3 mL), and the mixture was added to a sealed tube reactor. Cs2CO3 (411 mg, 1.26 mmol) was then added, and the mixture was reacted at 90 °C for 16 h. After the reaction was completed, the mixture was cooled to room temperature. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na2SO4, and filtered, and the filtrate was concentrated. The resulting crude product was subjected to silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8-fluoropyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01052] LC-MS: (ESI, m / z): [M+H]+ = 687.4.
[01053] Step 2: preparation of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)- 1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate TIPS
[01054] CataCXium A Pd-G3 (45 mg, 0.06 mmol) and cesium carbonate (299 mg, 0.92 mmol) were added to a mixed solution of tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-7-chloro-8- fluoropyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (210 mg, 0.31 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)naphthalen-1-yl)ethynyl)triisopropylsilane (188 mg, 0.37 mmol) in 1,4-dioxane (2 mL) and H2O (0.4 mL). The mixture was reacted at 85 °C for 16 h under nitrogen atmosphere. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give tert-butyl 3-(2-((R)-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01055] LC-MS: (ESI, m / z): [M / 2+H]+ = 519.5.
[01056] Step 3: preparation of tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8- ((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01057] tert-Butyl 3 -(2-((R )-3-(4-benzylpiperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3- (methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, 0.096 mmol) was dissolved in DCM (1 mL), and 1-chloroethyl chloroformate (41 mg, 0.29 mmol) and DIEA (37 mg, 0.29 mmol) were added. The mixture was reacted at room temperature for 1 h and concentrated under reduced pressure. MeOH (1 mL) was added to the concentrate for dissolution, and the mixture was heated to 50 °C and reacted for 10 min. After the reaction was completed, the mixture was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give tert-butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-((R)-2-methyl-3-(piperazin-1-yl)propoxy)pyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
[01058] LC-MS: (ESI, m / z): [M+H]+ = 948.5.
[01059] Step 4: preparation of tert-butyl 3-(2-((R)-3-(4-((3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3-azaspiro[5.5]undecan-9-yl)methyl)piperazin-1-yl)-2-methylpropoxy)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)pyrido[4,3-d]pyrimidin-4-yl)-1-((methoxy-d3)methyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate
[01060] Tetraisopropyl titanate (0.25 mL) was added to a stirred solution of tert -butyl 3-(8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-...
Claims
1. A compound of formula I, and / or a stereoisomer, an enantiomer, a diastereomer, an atropisomer, a deuteride, a hydrate, a solvate, or a prodrug thereof, and / or a pharmaceutically acceptable salt thereof:R1 is H or halogen;R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl;Ra is H, fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl, wherein the C1-3 alkyl, C1-3alkyl-O-C1-3 alkyl, C2-4 alkenyl, or C2-4 alkynyl is optionally substituted with 1 or more halogen, OH, or deuterium;Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1-3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;in G2,W is N or CRza;Rza is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy;Z is N or CRzb;Rzb is hydrogen, halogen, C1-6 alkyl, or C1-6 alkoxy;Rzc is phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole,, or pyridine, wherein the phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole,indazole,, or pyridine is optionally substituted with 1 or more R1';Rzd is hydrogen, deuterium, methyl, deuterated methyl, halomethyl, -CD2F, -CDF2, C2-6 alkyl, or C3-6 cycloalkyl, wherein the methyl, C2-6 alkyl, or C3-6 cycloalkyl is optionally substituted with 1 or more hydroxy, deuterium, or halogen;each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, -C(O)NR23R24, CH2R25, N=S(O)(C1-6 alkyl)2, S(O)C1-6 alkyl, S(O)2R26, -S-C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-6 hydroxyalkynyl, C1-6 cyanoalkyl, triazolyl, -S-C1-6 haloalkyl, C1-6 hydroxyalkyl, CH2C(O)NR27R28, -C2-6 alkynyl-NR29R30, C2-6 deuterated alkynyl, (C1-6 alkoxy)C1-6 haloalkyl-, or cycloalkyl, wherein the cycloalkyl is optionally substituted with 1 or more halogen or C1-6 alkyl;or R1' is SF5;R21 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C(O)C1-6 alkyl;R22 is hydrogen, C1-6 alkyl, or C1-6 haloalkyl;or R21 and R22, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;R23 and R24 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;or R23 and R24, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;R25 is hydroxy, cyano, heterocyclyl, NR25aR25b, C(O)NR25cR25d, or SO2C1-6 alkyl;R25a, R25b, R25c, and R25d are each independently H or C1-6 alkyl;or R25a and R25b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;or R23c and R25d, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;R26 is C1-6 alkyl, C1-6 haloalkyl, or NR26aR26b;R26a and R26b are each independently H or C1-6 alkyl;or R26a and R26b, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;R27 and R28 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;or R27 and R28, together with the nitrogen atom to which they are attached, form 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;R29 and R30 are each independently H, C1-6 alkyl, or C1-6 haloalkyl;or R29 and R30, together with the nitrogen atom to which they are attached, form 3- to 6-memberedheterocycloalkyl, wherein the 3- to 6-membered heterocycloalkyl is optionally substituted with 1 or more halogen, methyl, trifluoromethyl, methoxy, or trifluoromethoxy;R3a is hydrogen, halogen, hydroxy, cyano, C1-3 alkyl, C1-3 deuterated alkyl, or C1-3 haloalkyl;n is 0, 1, 2, 3, 4, or 5;,,,or, and L is linked to G via A or B;0o0,,,,0O , orCl 02. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof according to claim 1, which meets one or more of the following conditions:(1) X is N;(2) R1 is halogen;(3) R2 is C2-C4 alkynyl;(4) Ra is fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium; preferably, Ra is C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium;(5) in Ra, the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more deuterium;(6) Rb is H;(7) Rc is halogen;(10) W is CRza;(11) Rza is H or halogen, e.g., halogen;(12) Z is N;(13) Rzd is methyl, deuterated methyl, halomethyl, -CD2F, -CDF2, or C2-6 alkyl, wherein the C2-6 alkyl is optionally substituted with 1 or more hydroxy, deuterium, or halogen; preferably, Rzd is methyl or deuterated methyl;(14) n is 0;(15) Rzc is phenyl, naphthyl, or benzo[b]thiophene, wherein the phenyl, naphthyl, or benzo[b]thiophene is optionally substituted with 1 or more R1'.
3. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula I is selected from any one of the following schemes: scheme 1: in the compound of formula I,R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl;Ra is C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium;Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1-3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;L isR1 is halogen;orR2 is C2-C4 alkynyl;Ra is C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4alkenyl is optionally substituted with 1 or more deuterium;Rb is H;Rc is halogen;L iso A / . . . . Uffe A Y N 1 [ Xx , 0 a? ^° / \ N i । x\U , o a? yxN^y a ‘ ^N^qJ , 0 a? b a? / !<oxu / x'n'U u^-x^y , o a / . . . y"N^yB A / ^O N y I / c OMe L lx X / xX XX , or is linked to G via A; yy "fi CD3 OCD3 N^O L.NH L / NH l / NH E is 0 , 0 , ® , or scheme 3: in the compound of formula I, H fvSyRa R1 Rb x^X^r2 0 ifl X UI -X Rc OH G is G1 ; X is C or N; R1 is H or halogen; R2 is C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl; , x. ■XX......ci:.....' , 0 A / ^N^yB X-Nx^xk^O , 0 / XnYb ■Vx^ U 1 MXzO , 0 x\O^B N I [ ^xU ^NxX\x , 0 b U^xU OMel^N. Jx. J , T xx xx xx , and L uff y'^o 1NH 0 . ;Ra is fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more halogen or deuterium;Rb and Rc are independently H, OH, halogen, C1-3 alkyl, or C1-3 alkoxy, wherein the C1-3 alkyl or C1-3 alkoxy is optionally substituted with 1 or more halogen, OH, or deuterium;0 Az . . ^N^yB N i T • k / N. / L / L is X" , o az ^kXkB z ° / \ N । k'x.k ^N^qJ , O az . . pn^b y Z O / \ N 1 I |\X k N^;,^O , o A / ^''N^' / b A / Xq^'X k''xk x , 0 az ^kkAk / z o n k k^xk k / N\k'^° , 0 A / k^N^y B A / xoxP °' , 0 A / . . k^^B AX ZO Y N |Xxk OMe k^N^kxx , X z A / B , 0 A? k^’i’X^B ^° kk N ] k^xk k^N\xk^° , o A / I^N^k / B z ° / \ N 1 । ixk , 0 xx^. / \Ci^B '°'^k"N'^ k^\k k-N\k\ / O , 0 / XX^ / X x\X?^B ° k N । k^xk k / N^kx^0 , 0 % kkX^ b z ° k N । k^'-'-k k / N^kxX , kk N 1 kx\k X. k^N^kxx , 0 k'^'X^ B "O'^^X^N'^k k^X-k OMe k^N^kxx , 0 | k^xk FY k^N^kxx F ,B, and L is linked to G via A;E isorscheme 4: in the compound of formula I,>aOHG is G1X is N;R1 is halogen;R2 is C2-C4 alkynyl;Ra is fluorine, C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl, wherein the C1-3 alkyl, C1-3 alkyl-O-C1-3 alkyl, or C2-4 alkenyl is optionally substituted with 1 or more deuterium;Rb is H;Rc is halogen;, and L is linked, or,,;to G via A;E isscheme 5: in the compound of formula I,Rza is hydrogen or halogen;Z is N;Rzc is phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole, or pyridine, wherein the phenyl, naphthyl, quinoline, isoquinoline, benzo[b]thiophene, indole, indazole, or pyridine is optionally substituted with 1 or more R1';each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, C2-6 alkenyl, C2-6 alkynyl, C2-6 deuterated alkynyl, or cycloalkyl;R21 is hydrogen;R22 is hydrogen;Rzd is hydrogen, deuterium, methyl, deuterated methyl, halomethyl, -CD2F, -CDF2, or C2-6 alkyl, wherein theC2-6 alkyl is optionally substituted with 1 or more hydroxy, deuterium, or halogen;R3a is hydrogen, halogen, hydroxy, cyano, C1-3 alkyl, C1-3 deuterated alkyl, or C1-3 haloalkyl;n is 0 or 1;oris linked to G via A;G2G isW is CRza;Rza is halogen;Z is N;Rzc is phenyl, naphthyl, or benzo[b]thiophene, wherein the phenyl, naphthyl, or benzo[b]thiophene isoptionally substituted with 1 or more R1';each R1' is independently C1-6 alkyl, halogen, hydroxy, cyano, C1-6 alkoxy, C1-6 haloalkyl, C1-6 haloalkoxy, NR21R22, C2-6 alkenyl, C2-6 alkynyl, C2-6 deuterated alkynyl, or cycloalkyl;R21 is hydrogen;R22 is hydrogen;Rzd is methyl or deuterated methyl;n is 0;4. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, thedeuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereofaccording to claim 1, wherein G is:CD3OHOHOHOH5. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof according to claim 4, wherein the compound of formula I meets one or more of the following conditions:A I DVD V‘^'Cy, D dU A / VD□ / -, / / . 'aMX^, / / .,J -zX-^ ,° A / P ,, oroJ ZB,0,oN'Vb,0P^ / b / 0,0N^BJ / B, and E isV V, A9- aq, A?.m<N^° Pp° rNP° r^v°pNH >rNH ><NH Pnh0 , 0 , 0 , 0 , or O ;H JMl ^A PPp J]'N (2) G is OH , L is 0 A / B a / z".....^cccc- ' , 0 A / AA 7B A A 1 , A / Pq^ 0 , A pa rNY° rY Ay Anh Anh Anh Anf 0 0 o 0 ,,, H Apo- F III A Ax v, OH , L is O A / AN B '"'i.......o , 0 CAb '0"ta a 1 ,N ,1 ,0 , ’.....i.....-:i.....cP , P^N^P B , and E is a[A OCD3 0 'Y'0 1 A , or 0 ; preferably, G is O AA"" / ^ b Tp ‘ 'J orA / N^N "*W:y H (1 fo. f G is 0H or H M. / CjA (3) G is 0H o A / i^N^S '"•'Cl......C>.....1 H A J -.A (1 fY rz<fN / (4) G is oh , L is o A / A'N^ AkJ o A / A''rr Aq^y Aj\^ k / N^Y^O y0 y! ? rV rV 7 B Y"NH Ynh , and E is or ® ; more preferably, Y°" Ml A MM N|]N ^rVA OH • ; Acd3 o A / B , L is or yO / ^Ol AjA^Q / l^' jnY° zb a<nh >rNH , and E is 0 or 0 ; aa AnYb AAaYjn^b AoA^nA AMA ° / \ Y 1 (KJ 'M , , o B AZ 1^A B A]\Y , , 0 Ar AZ Y'N^" / b a.....q.......rj-.....1 , ,and E is00preferably,or;B,L is, and E isH e Nr py $ (8) G is OH or oh 0 A / y^ N B XX o A / Z^N^y B 'CXXI'X ypxp A / V r^N ^O'T nQ pp o a / i^N^y R CP ZB dD^ 0 A 7 DyD ^°'xy / x'N' / y y^xy vX / X / D D c A / % / D XD X^P y^xy X / Ny^ / D D 0 A / °VD 'X......X’ H 1 N ^ Z o fo / x Xy l v Xyx py Zb , L is ‘ '' , o A / B xy-xy yS\X , , o A / r^^N^y B y^xy \ / Nx / yo , , ? 0 Z B A / V a°tpn cP , , 0 A z DvD XD b Xtizx ■ , , B az dxd ^N^yB l^^N\XxX D D , , y b 9 AZV Zb Xc........cP , , o az y^N^J^yB xoX D D , ,OO^N•PC,H A / / Pb, or, and E isV. V- aq,.N..OOH'3N^° rNy° fNV°l^NH l^NH l^NH, 0 , 0 , 0 , orH HN^O rNH .O ; preferably, G is(q"Yor OHz O, L isI.......° .......1'0......cP0 o^.....O. aH AA n^o A is 0 ; Y'.y n ,scd3 Ij -J ni AL Y- 2:9-- (9) G is 0H or 0H , L is ? az .A Y< , o AZ A ANAb n ° A QA Gy D D A , A D o AA\ D D , o ...... rO"'A }......"■ 1 ANA\^o , ? A / yYxcP ' , 0 A7' —Ya , and e o AnYb 'nxY D D , o "aP’7' , 0 AnGb 9 PP "nA D 0 , 0 P^B a ^N^AO D D , 0 An^b A fAA 7 ^N^AO ,6. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, thedeuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereofaccording to claim 1, wherein G2 is G2-1, G2-2, G2-3, or G2-4:wherein Z, W, Rzc, Rzd, R3a, and n in G2-1, G2-2, G2-3, or G2-4 are as defined in any one of claims 1-3.
7. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereofaccording to claim 6, wherein G2 is G2-2-1, G2-2-2, or G2-2-3:G2-2-2wherein Rza, Rzb, Rzc, Rzd, R3a, and n in G2-2-1, G2-2-2, or G2-2-3 are as defined in claim 1.
8. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereofaccording to claim 7, wherein G2 is any one of the following schemes:scheme 1: G2 isNH2,,,nh2;scheme 2: G2 is:OH , OH , or OH9. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereofaccording to claim 1, wherein the compound of formula I is the following compound:P26413667AU428P26413667AUP26413667AUP26413667AUP26413667AUCl,the prodrug of the compound of formula I is any one of the following compounds:OxOOOMeO°yo / x a o' ° H2N „CN O 0F3 N^N Z\,-Vk )=( N '.■. ,„ rY V- S i V ' T OXr Yr fV i nh2 r । VH 0 , <° m3 y H2N CnO ° NN SuT F N^.0 F VH 0 , O^O^Yy- - 1' ' OH f" Y OH T 0 , °M °Y° *<N * ^1^0°3 OH ( Y OH VH o , r° ° Yr -f V Y. -h , A., --,, OH f Y OH VH 0 , O rN> 3Oy3 M\< / o zN-"7 A o ^O^QXPMoMe . N^O \NH O Oy_o rN N-^ ) O ' N^XUPV-e ' . N^O rNH O (° .N^CD3 Y o -^oyP'Va YH 0 ,0-3^ "-A.;: Y: V" 0 YH 010. The compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof according to claim 1, wherein the diastereomer has any one of the following structures:H H11. A pharmaceutical composition, comprising a therapeutically effective amount of the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof according to any one of claims 1-10, and a pharmaceutically acceptable carrier, diluent, or excipient.
12. Use of a substance X in the preparation of a medicament, wherein the substance X is the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof according to any one of claims 1-10, or the pharmaceutical composition according to claim 11; the medicament is for use as a medicament for treating or preventing a KRAS mutation-mediated disease or disorder, or for use as a medicament for treating or preventing a cancer;preferably, the KRAS mutation-mediated disease or disorder is a disease or disorder caused by the interaction between KRAS G12D and the SOS1 or SHP2 protein, or is a KRAS G12D mutation-mediated disease or disorder;the KRAS mutation-mediated disease or disorder and the cancer are independently selected from: heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, and liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma;lung: bronchial cancer (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, and adenocarcinoma), alveolar carcinoma (bronchioloalveolar carcinoma), bronchial adenoma, sarcoma, lymphoma, chondroma, hamartoma, and mesothelioma;gastrointestinal tract: esophagus cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, and lymphoma), stomach (cancer, lymphoma, and leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, and vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi’s sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, and fibroma), and large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, and leiomyoma);genitourinary system: kidney (adenocarcinoma, embryonal carcinosarcoma (Wilms’ tumor), lymphoma, and leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, and adenocarcinoma), prostate (adenocarcinoma and sarcoma), and testis (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, and lipoma);liver: liver cancer (hepatocellular carcinoma), intrahepatic cholangiocarcinoma, hepatoblastoma, malignant hemangioendothelioma, hepatocellular adenoma, and hemangioma;biliary tract: cholangiocarcinoma, gallbladder cancer, ampullary cancer, and intrahepatic cholangiocarcinoma; bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing’s sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, chondroma (osteocartilaginous exostosis), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor;nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, and Paget’s disease of bone), meninges (meningioma, meningeal sarcoma, and glioma), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinal cell tumor (pinealoma), glioblastoma multiforme, oligodendroglioma, neurilemmoma, eye cancer, and congenital tumor), spinal neurofibroma, meningioma, glioma, and sarcoma);gynecology and obstetrics: uterus (endometrial cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, and unclassified cancer), granulosa-theca cell tumor, ovarian Sertoli-Leydig cell tumor, dysgerminoma, and malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, and melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botulinum sarcoma (embryonal rhabdomyosarcoma), and fallopian tube (cancer);hematology: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disease, multiple myeloma, and myelodysplastic syndrome), Hodgkin’s disease, and non-Hodgkin’s lymphoma (malignant lymphoma);skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi’s sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, and psoriasis; and adrenal gland: neuroblastoma;for example, the KRAS mutation-mediated disease or disorder and the cancer are independently selected from: pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial cancer, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostatecancer, glioblastoma, renal cancer, and sarcoma;for another example, the cancer and the KRAS mutation-mediated disease or disorder are independently selected from gastric cancer, pancreatic cancer, pancreatic adenocarcinoma, and colon cancer.
13. A method for treating or preventing a cancer or a KRAS G12D-mediated disease or disorder, comprising administering to a patient in need thereof a therapeutically effective amount of a substance X, wherein the substance X is the compound of formula I, and / or the stereoisomer, the enantiomer, the diastereomer, the atropisomer, the deuteride, the hydrate, the solvate, or the prodrug thereof, and / or the pharmaceutically acceptable salt thereof according to any one of claims 1-10, or the pharmaceutical composition according to claim 11;preferably, the cancer and the KRAS G12D-mediated disease or disorder are as described in claim 12.