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WO2024188246A1 - Composé macrocyclique contenant de l'azote, son procédé de préparation et son utilisation - Google Patents

Composé macrocyclique contenant de l'azote, son procédé de préparation et son utilisation Download PDF

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Publication number
WO2024188246A1
WO2024188246A1 PCT/CN2024/081244 CN2024081244W WO2024188246A1 WO 2024188246 A1 WO2024188246 A1 WO 2024188246A1 CN 2024081244 W CN2024081244 W CN 2024081244W WO 2024188246 A1 WO2024188246 A1 WO 2024188246A1
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alkyl
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substituted
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WO2024188246A9 (fr
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周国强
张诚一
张少敏
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Nutshell Biotech Shanghai Co Ltd
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Nutshell Biotech Shanghai Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the invention relates to a nitrogen-containing macrocyclic compound and a preparation method and application thereof.
  • Mammalian cells have evolved multiple pathways to repair DNA double-strand breaks (DSBs) to ensure genome stability.
  • the most studied mechanisms for repairing DNA double-strand breaks include non-homologous end joining (NHEJ) and homologous recombination (HR).
  • NHEJ does not require a homologous template or only requires a small amount of homologous template, and connects the DNA ends by inserting and deleting a few bases at the break site (Chang HHY, Pannunzio NR, Adachi N, Lieber MR. Non-homologous DNA end joining and alternative pathways to double-strand break repair. Nat Rev Mol Cell Biol.
  • DNA polymerase ⁇ (Pol ⁇ , encoded by POLQ) is a multifunctional DNA polymerase of the A family and has been identified as a key protein of the MMEJ pathway (Kent T et al. Mechanism of microhomology-mediated end-joining promoted by human DNA polymerase ⁇ . Nat Struct Mol Biol. 2015, 22(3): 230-7; Mateos-Gomez PA et al. Mammalian polymerase ⁇ promotes alternative NHEJ and suppresses recombination. Nature. 2015, 518(7538): 254-257).
  • POL ⁇ contains three domains and is structurally and functionally different from other polymerases (Ozdemir AY et al.
  • Homologous-recombination-deficient tumours are dependent on Pol ⁇ -mediated repair. Nature. 2015, 518(7538): 258-62; Mateos-Gomez PA et al. Mammalian polymerase ⁇ promotes alternative NHEJ and suppresses recombination. Nature. 2015, 518(7538): 254-7; Mateos-Gomez PA et al. The helicase domain of Pol ⁇ counteracts RPA to promote alt-NHEJ. Nat Struct Mol Biol. 2017, 24(12):1116-1123).
  • the C-terminus contains a nuclease domain that trims the DNA end and an error-prone polymerase domain that fills in nucleotides during MMEJ repair (Zahn KE, Jensen RB, Wood RD, D.00é S. Human DNA polymerase ⁇ harbors DNA end-trimming activity critical for DNA repair. Mol Cell. 2021, 81(7): 1534-1547). Both the ATPase domain and the polymerase domain are required for MMEJ repair (Beagan K et al. Drosophila DNA polymerase theta utilizes both helicase-like and polymerase domains during microhomology-mediated end joining and interstrand crosslink repair. PLoS Genet. 2017, 13(5): e1006813). Therefore, inhibiting Pol ⁇ activity and thereby depriving cells of MMEJ repair ability provides a new targeted strategy for treating certain specific tumors.
  • DNA polymerase ⁇ (Pol ⁇ ) is not essential in healthy cells, but its expression is upregulated in breast, lung and ovarian cancers, and its overexpression is associated with HR repair defects and poor clinical outcomes (Ceccaldi R et al. Homologous-recombination-deficient tumors are dependent on Pol ⁇ -mediated repair. Nature. 2015, 518(7538): 258-62).
  • HR deficiency loss of POLQ has been shown to impair cell viability, revealing a synthetic lethal relationship between POLQ and HR factors (e.g., POLQ and FA/BRCA deficiency are synthetically lethal) (Shima N, Munroe RJ, Schimenti JC.
  • the mouse genomic instability mutation chaos1 is an allele of Polq that exhibits genetic interaction with Atm. Mol Cell Biol. 2004, 24(2 3):10381-9; Wang Z et al., DNA polymerase ⁇ (POLQ) is important for repair of DNA double-strand breaks caused by fork collapse.J Biol Chem.2019,294(11):3909-3919; Ceccaldi R et al.Homologous-recombination-defic ient tumors are dependent on Pol ⁇ -mediated repair.Nature.2015,518(7538):258-62;Mateos-Gomez PA et al.Mammalian polymerase ⁇ promotes alternative NHEJ and suppresses recombination.Nature.2015,518(7538):254-7;Dai CH et al.Co-inhibition of pol ⁇ and HR genes efficiently synergi ze with cisplatin to suppress cisplatin-resistant lung cancer
  • Pol ⁇ is an attractive new target for synthetic lethal therapy, and the development of small molecule inhibitors of DNA polymerase ⁇ is of great significance.
  • the present invention provides a nitrogen-containing macrocyclic compound and a preparation method and application thereof.
  • the compound of the present invention has better inhibitory activity on DNA polymerase ⁇ .
  • the present invention provides a compound as shown in Formula I, a pharmaceutically acceptable salt thereof, an isotope-labeled substance thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a prodrug thereof:
  • L 1a , L 1b , L 2a and L 2b are independently H, deuterium, halogen, cyano, hydroxyl, -NL 1a-1 L 1a-2 , C 1-6 alkyl, C 1-6 alkoxy or C 3-6 cycloalkyl, and L 1a and L 1b are not H at the same time;
  • any adjacent L 1a together with the atoms to which they are connected form a C 3-6 cycloalkyl or a 3-6 membered heterocyclic group, or L 1a and L 1b together with the atoms to which they are commonly connected form a C 3-6 cycloalkane or a 3-6 membered heterocyclic ring;
  • the heteroatoms in the 3-6 membered heterocyclic group and the 3-6 membered heterocyclic ring are independently selected from 1, 2 or 3 of N, O and S;
  • L 1a-1 , L 1a-2 and L 3 are independently H, C 1-6 alkyl, C 1-6 alkyl substituted with one or more deuterium atoms, or C 3-6 cycloalkyl;
  • R 1 is H, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 alkyl substituted by one or more R 1-1 , or C 3-6 cycloalkyl substituted by one or more R 1-2 ;
  • R 1-1-1 , R 1-1-2 , R 1-1-3 , R 1-1-4 , R 1-1-5 , R 1-1-6 and R 1-1-7 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • R 2 , R 3a and R 3b are independently H or C 1-6 alkyl
  • R2 and R3a together with the atoms to which they are attached form a 5-6 membered heterocyclic group or a 5-6 membered heterocyclic group substituted by one or more R2-1 ;
  • the heteroatoms in the 5-6 membered heterocyclic group and the 5-6 membered heterocyclic group substituted by one or more R2-1 are independently selected from 1, 2 or 3 of N, O and S, and contain at least 1 N;
  • any two adjacent R 2-1's together with the atom to which they are attached form a ring A, wherein the ring A is a C 6-18 aryl group, a 5-10-membered heteroaryl group, a C 3-6 cycloalkyl group, a 5-10-membered heterocyclyl group, a C 6-18 aryl group substituted by one or more R 2-1-3 , a 5-10-membered heteroaryl group substituted by one or more R 2-1-4 , a C 3-6 cycloalkyl group substituted by one or more R 2-1-5 , or a 5-10-membered heterocyclyl group substituted by one or more R 2-1-6 ;
  • heteroatoms in the 5-10 membered heteroaryl, the 5-10 membered heterocyclic group, the 5-10 membered heteroaryl substituted by one or more R 2-1-4 , and the 5-10 membered heterocyclic group substituted by one or more R 2-1-6 are independently selected from 1, 2 or 3 of N, O and S;
  • R 2-1-1 and R 2-1-2 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • R 2-1-1-1 and R 2-1-1-2 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • Ar1 and Ar2 are independently C6-18 aryl, 5-10 membered heteroaryl, C6-18 aryl substituted by one or more R4-1 , 5-10 membered heteroaryl substituted by one or more R4-2, 5-10 membered heteroaryl and 3-8 membered heterocyclyl, 5-10 membered heteroaryl and 3-8 membered heterocyclyl substituted by one or more R4-2 (preferably, Ar1 and Ar2 are independently C6-18 aryl, 5-10 membered heteroaryl, C6-18 aryl substituted by one or more R4-1 , or 5-10 membered heteroaryl substituted by one or more R4-2 ) ;
  • heteroatoms in the 5-10 membered heteroaryl, the 5-10 membered heteroaryl substituted by one or more R 4-2 , the 5-10 membered heteroaryl and 3-8 membered heterocyclic group, and the 5-10 membered heteroaryl and 3-8 membered heterocyclic group substituted by one or more R 4-2 are independently selected from 1, 2 or 3 of N, O and S;
  • any two adjacent R 4-1's together with the atoms to which they are attached form a 5-10 heteroaryl, a C 3-6 cycloalkyl, a 5-10 membered heterocycloalkyl, a 5-10 membered heteroaryl substituted by one or more R 4-1-10 , a C 3-6 cycloalkyl substituted by one or more R 4-1-11 , or a 5-10 membered heterocycloalkyl substituted by one or more R 4-1-12 ;
  • heteroatoms in the 5-10 membered heteroaryl, 5-10 membered heterocycloalkyl, the 5-10 membered heteroaryl substituted by one or more R 4-1-10 , and the 5-10 membered heterocycloalkyl substituted by one or more R 4-1-12 are independently selected from 1, 2 or 3 of N, O and S;
  • R 4-1-1 , R 4-1-2 , R 4-1-3 , R 4-1-4 , R 4-1-5 , R 4-1-6 and R 4-1-7 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • R 4-1-8-1 , R 4-1-8-2 , R 4-1-8-3 , R 4-1-8-4 , R 4-1-8-5 , R 4-1-8-6 and R 4-1-8-7 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl.
  • L 1a , L 1b , L 2a and L 2b are independently H, deuterium, halogen, cyano, hydroxyl, -NL 1a-1 L 1a-2 , C 1-6 alkyl, C 1-6 alkoxy or C 3-6 cycloalkyl, and L 1a and L 1b are not H at the same time;
  • any adjacent L 1a together with the atoms to which they are connected form a C 3-6 cycloalkyl or a 3-6 membered heterocyclic group, or L 1a and L 1b together with the atoms to which they are commonly connected form a C 3-6 cycloalkane or a 3-6 membered heterocyclic ring;
  • the heteroatoms in the 3-6 membered heterocyclic group and the 3-6 membered heterocyclic ring are independently selected from 1, 2 or 3 of N, O and S;
  • L 1a-1 , L 1a-2 and L 3 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • R 1 is H, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 alkyl substituted by one or more R 1-1 , or C 3-6 cycloalkyl substituted by one or more R 1-2 ;
  • R 1-1-1 , R 1-1-2 , R 1-1-3 , R 1-1-4 , R 1-1-5 , R 1-1-6 and R 1-1-7 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • R 2 , R 3a and R 3b are independently H or C 1-6 alkyl
  • R2 and R3a together with the atoms to which they are attached form a 5-6 membered heterocyclic group or a 5-6 membered heterocyclic group substituted by one or more R2-1 ;
  • the heteroatoms in the 5-6 membered heterocyclic group and the 5-6 membered heterocyclic group substituted by one or more R2-1 are independently selected from 1, 2 or 3 of N, O and S, and contain at least 1 N;
  • any two adjacent R 2-1's together with the atom to which they are attached form a ring A, wherein the ring A is a C 6-18 aryl group, a 5-10-membered heteroaryl group, a C 3-6 cycloalkyl group, a 5-10-membered heterocyclyl group, a C 6-18 aryl group substituted by one or more R 2-1-3 , a 5-10-membered heteroaryl group substituted by one or more R 2-1-4 , a C 3-6 cycloalkyl group substituted by one or more R 2-1-5 , or a 5-10-membered heterocyclyl group substituted by one or more R 2-1-6 ;
  • heteroatoms in the 5-10 membered heteroaryl, the 5-10 membered heterocyclic group, the 5-10 membered heteroaryl substituted by one or more R 2-1-4 , and the 5-10 membered heterocyclic group substituted by one or more R 2-1-6 are independently selected from 1, 2 or 3 of N, O and S;
  • R 2-1-1 and R 2-1-2 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • each R 2-1-3 , each R 2-1-4 , each R 2-1-5 and each R 2-1-6 are independently deuterium, halogen, cyano, hydroxy, -NR 2-1-3-1 R 2-1-3-2 , C 1-6 alkyl, C 1-6 alkoxy or C 3-6 cycloalkyl;
  • R 2-1-1-1 and R 2-1-1-2 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • Ar1 and Ar2 are independently C6-18 aryl, 5-10 membered heteroaryl, C6-18 aryl substituted by one or more R4-1 , or 5-10 membered heteroaryl substituted by one or more R4-2 ;
  • heteroatoms in the 5-10 membered heteroaryl and the 5-10 membered heteroaryl substituted by one or more R 4-2 are independently selected from 1, 2 or 3 of N, O and S;
  • any two adjacent R 4-1's together with the atoms to which they are attached form a 5-10 heteroaryl, a C 3-6 cycloalkyl, a 5-10 membered heterocycloalkyl, a 5-10 membered heteroaryl substituted by one or more R 4-1-10 , a C 3-6 cycloalkyl substituted by one or more R 4-1-11 , or a 5-10 membered heterocycloalkyl substituted by one or more R 4-1-12 ;
  • heteroatoms in the 5-10 membered heteroaryl, 5-10 membered heterocycloalkyl, the 5-10 membered heteroaryl substituted by one or more R 4-1-10 , and the 5-10 membered heterocycloalkyl substituted by one or more R 4-1-12 are independently selected from 1, 2 or 3 of N, O and S;
  • R 4-1-1 , R 4-1-2 , R 4-1-3 , R 4-1-4 , R 4-1-5 , R 4-1-6 and R 4-1-7 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl;
  • R 4-1-8-1 , R 4-1-8-2 , R 4-1-8-3 , R 4-1-8-4 , R 4-1-8-5 , R 4-1-8-6 and R 4-1-8-7 are independently H, C 1-6 alkyl or C 3-6 cycloalkyl.
  • the halogen independently may be fluorine, chlorine or bromine, for example fluorine.
  • the C 1-6 alkyl group independently and the C 1-6 alkyl group substituted with one or more deuterates can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.
  • the C 1-6 alkoxy group in L 1a , L 1b , L 2a and L 2b , can independently be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy.
  • the C 3-6 cycloalkyl group may independently be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
  • L 1a and L 1b together with the atoms to which they are attached form a C 3-6 cycloalkane which may be cyclopropane, cyclobutane, cyclopentane or cyclohexane, for example, cyclopropane ( # indicates that a spiro ring is formed through this carbon atom).
  • the C 1-6 alkyl group substituted with one or more deuterium radicals may be -CD 3 .
  • the C1-6 alkyl group and the C1-6 alkyl group substituted by one or more R1-1 can independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.
  • the C 1-6 alkyl group substituted by one or more R 1-1 may be a C 1-6 alkyl group substituted by 1, 2 or 3 deuterium groups, such as -CD 3 .
  • the C 1-6 alkyl group in R 2 , R 3a and R 3b , can independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.
  • R2 and R3a together with the atoms to which they are attached form the 5-6 membered heterocyclic group and the 5-6 membered heterocyclic group substituted by one or more R2-1 can independently be pyrrolidinyl, for example It is connected to Ar 1 through N and to the carbonyl group through C.
  • the C 1-6 alkoxy group in each R 2-1 , each R 2-1-3 , each R 2-1-4 , each R 2-1-5 and each R 2-1-6 can independently be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy.
  • each R 2-1 , R 2-1-1 , R 2-1-2 , each R 2-1-3 , each R 2-1-4 , each R 2-1-5 , each R 2-1-6 , R 2-1-1-1 and R 2-1-1-2 , the C 1-6 alkyl group can independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.
  • any two adjacent R 2-1's together with the atoms to which they are connected form the C 6-18 aryl group and the C 6-18 aryl group substituted by one or more R 2-1-3 's can independently be phenyl or naphthyl.
  • the sub-group is selected from N, and the number is 1 or 2, for example, pyridyl Pyrazolyl or pyrimidine This means that a cyclic structure is formed with a 5- to 6-membered heterocyclic group through the bond therein.
  • the C 6-18 aryl group and the C 6-18 aryl group substituted by one or more R 4-1 can independently be phenyl or naphthyl, for example phenyl.
  • the 5-10 membered heteroaryl and the 5-10 membered heteroaryl substituted by one or more R4-2 can be independently a 5-10 membered heteroaryl, wherein the heteroatom is selected from N, and the number of the heteroaryl is 1 or 2, and can also be a 5-6 membered monocyclic heteroaryl or an 8-10 membered cyclic heteroaryl, such as pyridyl or (Preferably pyridyl).
  • the 5-10-membered heteroaryl and 3-8-membered heterocyclic group and the 5-10-membered heteroaryl and 3-8-membered heterocyclic group substituted by one or more R4-2 can be independently a 5-6-membered heteroaryl and 3-6-membered heterocyclic group, wherein the heteroatom is selected from N, the number of which is 1 or 2 , and can also be a pyridyl and pyrrolidinyl group, for example
  • the C 1-6 alkyl group and the C 1-6 alkyl group substituted by one or more R 4-1-8 can independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.
  • the C 1-6 alkoxy group and the C 1-6 alkoxy group substituted by one or more R 4-1-9 can independently be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy.
  • any two adjacent R 4-1 and the atoms to which they are connected together form the 5-10-membered heteroaryl and the 5-10-membered heteroaryl in the 5-10-membered heteroaryl substituted by one or more R 4-1-10 can be a 5-6-membered heteroaryl, wherein the heteroatom is selected from N, and the number is 1 or 2, for example, pyrazolyl ( It means that a ring structure is formed with a C 6-18 aryl group through the bond here).
  • the C 1-6 alkyl group can independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.
  • the C 3-6 cycloalkyl group may independently be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
  • each R 4-1 , each R 4-2 , each R 4-1-8 , each R 4-1-9 , each R 4-1-10 and each R 4-1-11 can independently be fluorine, chlorine or bromine, for example fluorine or chlorine.
  • each R 4-1 , each R 4-2 , each R 4-1-8 , each R 4-1-9 , each R 4-1-10 and each R 4-1-11 can independently be fluorine, chlorine or bromine, for example fluorine or chlorine.
  • the C 1-6 alkyl substituted by one or more R 4-1-8 may be a C 1-6 alkyl substituted by 1, 2 or 3 halogens, such as -CF 3 .
  • one of L 1a and L 1b is H, and the other is halogen, hydroxyl or C 1-6 alkyl; or L 1a and L 1b together with the atoms to which they are attached form a C 3-6 cycloalkane or a 3-6 membered heterocycle.
  • L 3 is H, C 1-6 alkyl, or C 1-6 alkyl substituted with one or more deuteriums.
  • -CL 1a L 1b - e.g., -CH(CH 3 )-, -CH(OH)-, -CHF 2 - or
  • -NL 3 - e.g., -N(CH 3 )- or -N(CD 3 )-
  • the -(CH 2 ) n - may be any one or two -CH 2 - in -(CH 2 ) n - replaced by one or two of -CL 1a L 1b -
  • -CL 2a CL 2b -, -NL 3 -, -O-, -S
  • -L- contains 3, 4 or 5 atoms in the main chain, such as 3-4 atoms, for example 3 atoms.
  • R 1 may be H or a C 1-6 alkyl group (eg, methyl) or a C 1-6 alkyl group substituted by one or more R 1-1 groups (eg, -CD 3 ).
  • R 2 , R 3a and R 3b are defined as in case 1 or 2:
  • R 2 , R 3a and R 3b are H or C 1-6 alkyl, or R 2 and R 3a together with the atoms to which they are attached form a 5-6 membered heterocyclic group or a 5-6 membered heterocyclic group substituted by one or more R 2-1 ;
  • any two adjacent R 2-1 and the atoms to which they are attached together form a ring A, wherein the ring A is a 5-10 membered heteroaryl, or or a 5-10 membered heteroaryl group substituted by one or more R 2-1-4 ;
  • R 2 , R 3a and R 3b are H or C 1-6 alkyl, or R 2 and R 3a together with the atoms to which they are attached form a 5-6 membered heterocyclic group or a 5-6 membered heterocyclic group substituted by one or more R 2-1 ;
  • any two adjacent R 2-1's and the atoms to which they are connected together form a ring A, wherein the ring A is a 5-10 membered heteroaryl group, or a 5-10 membered heteroaryl group substituted by one or more R 2-1-4 ;
  • Each R 2-1-4 is independently a C 1-6 alkyl group.
  • the C atom marked with * can be a chiral carbon atom or an achiral carbon atom; when the C atom marked with * is a chiral carbon atom, the configuration of the chiral carbon atom is S configuration;
  • the configuration of the chiral carbon atom is S configuration.
  • Ar 1 can be a 5-10 membered heteroaryl group, or a 5-10 membered heteroaryl group substituted by one or more R 4-2 ;
  • Each R 4-2 is independently cyano, C 1-6 alkyl, or C 1-6 alkyl substituted by one or more R 4-1-8 ;
  • Each R 4-1-8 is independently halogen.
  • Ar 1 may be (Preferably )(Ar 1 above Connected to the N atom, the following Connected to L (with fragment For example, according to the reading habit from top to bottom, mark position a as the top and position b as the bottom)).
  • Ar 2 may be a C 6-18 aryl group, or a C 6-18 aryl group substituted by one or more R 4-1 ;
  • Each R 4-1 is independently C 1-6 alkyl, C 1-6 alkyl substituted with one or more deuterated radicals, or halogen, or any two adjacent R 4-1 together with the atoms to which they are attached form a 5-10 membered heteroaryl or a 5-10 membered heteroaryl substituted with one or more R 4-1-10 ;
  • Each R 4-1-10 is independently a C 1-6 alkyl group.
  • each R 4-1 is independently C 1-6 alkyl or halogen, or any two adjacent R 4-1 together with the atom to which they are attached form a 5-10 membered heteroaryl or a 5-10 membered heteroaryl substituted by one or more R 4-1-10 .
  • Ar 2 may be (Ar 2 above Connected to the N atom, the following connected to L).
  • the number of ring atoms in the ring may be 11, 12 or 13, for example 12.
  • the compound as shown in Formula I can be any one of the general formulas I-1 to I-7:
  • n1 and n2 are independently 0, 1, 2 or 3; Z is C or N (for example, Z is C); L, R 1 , R 2 , R 3a , R 3b , R 4-1 and R 4-12 are defined as above;
  • Y is CHR 2-1 or C(R 2-1 )(R 2-1 ); n2 is 0, 1 or 2; L, R 1 , R 2-1 , R 3b , Ar 1 and Ar 2 are defined as above;
  • Y is N, CH or CHR 2-1 ; n3 is 0, 1 or 2; ring A, L, R 1 , R 2-1 , R 3b , Ar 1 and Ar 2 are defined as above;
  • Y is N, CH or CHR 2-1 ; n4 is 0, 1 or 2; ring A, L, R 1 , R 2-1 , R 3b , Ar 1 and Ar 2 are defined as above;
  • R 2 , R 3a and R 3b are independently H or C 1-6 alkyl; L, R 1 , Ar 1 and Ar 2 are defined as above;
  • n3 is independently 0, 1, 2 or 3;
  • L, R 1 , R 2-1 , R 3b , Ar 1 and Ar 2 are defined as above;
  • n4 is independently 0, 1, 2 or 3;
  • Ring B is a 3-8 membered heterocycle (e.g., a 3-6 membered heterocycle, and another example is a pyrrole ring) or a 5-6 membered heteroaromatic ring (e.g., a 5 membered heteroaromatic ring, the heteroatom is N, and another example is an imidazole ring), and the heteroatoms in the 3-8 membered heterocycle and the 5-6 membered heteroaromatic ring are independently selected from 1, 2 or 3 of N, O and S;
  • L, R 1 , R 2-1 , R 3b , Ar 1 and Ar 2 are all defined as above.
  • Ar 1 is a pyridyl group substituted by one or more R 4-2 (such as ), Ar 2 is a phenyl group substituted by one or more R 4-1 (such as
  • R 1 is H, C 1-6 alkyl, or C 1-6 alkyl substituted by one or more R 1-1 ;
  • R 2 , R 3a and R 3b are H or C 1-6 alkyl, or R 2 and R 3a together with the atoms to which they are attached form a 5-6 membered heterocyclic group or a 5-6 membered heterocyclic group substituted by one or more R 2-1 ;
  • Each R 2-1-4 is independently a C 1-6 alkyl group
  • Ar 1 is a 5-10 membered heteroaryl group, or a 5-10 membered heteroaryl group substituted by one or more R 4-2 ;
  • Each R 4-2 is independently cyano, C 1-6 alkyl, or C 1-6 alkyl substituted by one or more R 4-1-8 ;
  • Each R 4-1-8 is independently halogen
  • Ar 2 is a C 6-18 aryl group, or a C 6-18 aryl group substituted by one or more R 4-1 ;
  • Each R 4-1 is independently C 1-6 alkyl or halogen, or any two adjacent R 4-1 together with the atom to which they are attached form a 5-10 membered heteroaryl or a 5-10 membered heteroaryl substituted by one or more R 4-1-10 ;
  • Each R 4-1-10 is independently a C 1-6 alkyl group.
  • the compound as shown in formula I is preferably any of the following compounds:
  • the compound as shown in formula I is any of the following compounds:
  • the present invention also provides a method for preparing the compound of formula I, a pharmaceutically acceptable salt thereof, an isotope-labeled substance thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a prodrug thereof, wherein the method for preparing the compound of formula I is any one of methods 1 to 8:
  • Method 1 comprises the following steps: reacting a compound as shown in Formula II-1 in a solvent in the presence of a condensing agent or a base (such as lithium bis(trimethylsilyl)amide) to obtain the compound as shown in Formula I;
  • R 5 is hydrogen or C 1-6 alkyl
  • R 2 , R 3a , R 3b , Ar 1 , Ar 2 and L are as defined above;
  • R 1 is C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 alkyl substituted by one or more R 1-1 , or C 3-6 cycloalkyl substituted by one or more R 1-2
  • the preparation method of the compound of formula I is method 2;
  • Method 2 comprises the following steps: in the presence of a base (such as NaH), a compound as shown in formula II-2 and compound III are mixed in a solvent. Carry out a substitution reaction to obtain the compound shown in formula I;
  • a base such as NaH
  • X is a halogen (eg, iodine);
  • R 2 , R 3a , R 3b , Ar 1 , Ar 2 and L are as defined above;
  • Method 3 comprises the following steps: subjecting the compound represented by formula II-3 to an intramolecular Mitsunobu reaction in a solvent (such as reacting in the presence of triphenylphosphine and diisopropyl azodicarboxylate) to obtain the compound represented by formula I;
  • R 1 , R 2 , R 3a , R 3b , Ar 1 and Ar 2 are as defined above;
  • Method 4 comprises the following steps: subjecting the compound represented by formula II-4 to an intramolecular Mitsunobu reaction in a solvent (such as reacting in the presence of triphenylphosphine and diisopropyl azodicarboxylate) to obtain the compound represented by formula I;
  • R 1 , R 2 , R 3a , R 3b , Ar 1 and Ar 2 are as defined above;
  • Method 5 comprises the following steps: in the presence of a catalyst (such as a Grubbs third generation catalyst), subjecting the compound represented by formula II-5 to an intramolecular olefin metathesis reaction in a solvent to obtain the compound represented by formula I;
  • a catalyst such as a Grubbs third generation catalyst
  • n5 and n6 are independently 0, 1, 2, 3 or 4, and the sum of n5 and n6 is less than or equal to 5;
  • R 1 , R 2 , R 3a , R 3b , Ar 1 and Ar 2 are as defined above;
  • Method 6 comprises the following steps: in the presence of a catalyst (such as Pd/C) and hydrogen, subjecting the compound represented by formula II-6 to a reduction reaction in a solvent to obtain the compound represented by formula I;
  • a catalyst such as Pd/C
  • L 4 is C 2-6 alkenyl (e.g., propenyl); R 1 , R 2 , R 3a , R 3b , Ar 1 and Ar 2 are as defined above;
  • the preparation method of the compound of formula I is method 7;
  • Method 7 comprises the following steps: in the presence of an oxidant (such as ruthenium trichloride hydrate and sodium periodate), subjecting the compound represented by formula II-7 to an oxidation reaction in a solvent to obtain the compound represented by formula I;
  • an oxidant such as ruthenium trichloride hydrate and sodium periodate
  • n7 is 1, 2 or 3;
  • R 3a , R 3b , Ar 1 and Ar 2 are as defined above;
  • Method 8 comprises the following steps: in the presence of an acid (hydrochloric acid), subjecting the compound represented by formula II-8 to a deprotection reaction in a solvent to obtain the compound represented by formula I;
  • M is a hydroxyl protecting group (such as tert-butyldimethylsilyl (-TBS)); R 2-1 , R 3a , R 3b , Ar 1 and Ar 2 are as defined above.
  • R 2-1 , R 3a , R 3b , Ar 1 and Ar 2 are as defined above.
  • the present invention also provides a compound as shown in formula II-1 to formula II-7, II-8-1 to II-8-2 (the compounds as shown in formula II-1 to formula II-7, II-8-1 to II-8-2 can be used to prepare the above-mentioned compound as shown in formula I):
  • n5, n6, n7, R 1 , R 2 , R 2-1 , R 3a , R 3b , Ar 1 , Ar 2 , L, L 4 and M are as defined above.
  • the formula II-1 is preferably
  • the formula II-2 is preferably
  • the formula II-3 is preferably any of the following compounds:
  • the formula II-5 is preferably:
  • the formula II-6 is preferably
  • the formula II-7 is preferably
  • the formula II-8 is preferably
  • the present invention also provides a compound:
  • the present invention also provides a pharmaceutical composition, which includes the above-mentioned compound as shown in Formula I, its pharmaceutically acceptable salt, its isotope-labeled substance, its solvate, its pharmaceutically acceptable salt solvate or its prodrug, and pharmaceutical excipients.
  • the amount of the compound as shown in Formula I, its pharmaceutically acceptable salt, its isotope-labeled substance, its solvate, its pharmaceutically acceptable salt solvate or its prodrug can be an effective therapeutic amount.
  • the present invention also provides a use of the above-mentioned compound as shown in Formula I, its pharmaceutically acceptable salt, its isotope-labeled substance, its solvate, its solvate of a pharmaceutically acceptable salt or its prodrug in the preparation of a DNA polymerase ⁇ (Pol ⁇ ) inhibitor (in vivo or in vitro (for example, for laboratory experiments)).
  • a DNA polymerase ⁇ (Pol ⁇ ) inhibitor in vivo or in vitro (for example, for laboratory experiments)
  • the present invention also provides a use of the above-mentioned compound as shown in Formula I, its pharmaceutically acceptable salt, its isotope-labeled product, its solvate, its pharmaceutically acceptable salt solvate or its prodrug in the preparation of a drug.
  • the drug is used to prevent or treat cancers selected from the group consisting of ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, colorectal cancer, lung cancer and cervical cancer.
  • the present invention also provides the use of the above-mentioned compound as shown in Formula I, its pharmaceutically acceptable salt, its isotope-labeled product, its solvate, its solvate of a pharmaceutically acceptable salt or its prodrug in the preparation of a drug for treating or preventing diseases associated with DNA repair defects.
  • the disease associated with DNA repair deficiency may be the above-mentioned cancer.
  • the present invention also provides the use of the above-mentioned compound as shown in Formula I, its pharmaceutically acceptable salt, its isotope-labeled product, its solvate, its pharmaceutically acceptable salt solvate or its prodrug in the preparation of a drug for treating or preventing the above-mentioned cancer.
  • the present invention also provides a method for treating and/or preventing a disease characterized by overexpression of DNA polymerase ⁇ in a patient, which comprises administering to the patient a therapeutically effective amount of the compound as shown in Formula I, a pharmaceutically acceptable salt thereof, an isotope-labeled substance thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a prodrug thereof.
  • the present invention also provides a method for treating and/or preventing diseases associated with DNA repair defects, which comprises administering to a patient a therapeutically effective amount of the above-mentioned compound as shown in Formula I, a pharmaceutically acceptable salt thereof, an isotope-labeled substance thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a prodrug thereof.
  • the disease associated with DNA repair deficiency may be the above-mentioned cancer.
  • the present invention also provides a method for treating and/or preventing the above-mentioned cancer, which comprises administering to a patient a therapeutically effective amount of the above-mentioned compound as shown in Formula I, a pharmaceutically acceptable salt thereof, an isotope-labeled substance thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a prodrug thereof.
  • a group B substituted by 1 or more groups A means 2, 3, 4 or 5.
  • the expression "a group B substituted by 1 or more groups A” means that 1, 2, 3, 4 or 5 hydrogen atoms in the group B are independently replaced by groups A.
  • groups A When multiple A groups appear at the same time, unless otherwise specified, their definitions are independent of each other and do not affect each other.
  • a C 6 -C 10 aryl substituted by 3 halogens means that a C 6 -C 10 aryl group will be substituted by 3 halogens, and the definitions of the 3 halogens are independent of each other and do not affect each other, including but not limited to: wait.
  • pharmaceutically acceptable means relatively non-toxic, safe, and suitable for use by patients.
  • pharmaceutically acceptable salt refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base.
  • a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent.
  • an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent.
  • solvate refers to a substance formed by the combination of a compound and a solvent (including but not limited to: water, methanol, ethanol, etc.). Solvates are divided into stoichiometric solvates and non-stoichiometric solvates.
  • pharmaceutically acceptable salt solvate refers to a substance formed by the combination of a compound with a pharmaceutically acceptable acid or base and a solvent (including but not limited to water, methanol, ethanol, etc.), wherein the amount of the solvent may be stoichiometric or non-stoichiometric.
  • the structural fragment is connected to the rest of the molecule through this site.
  • It refers to cyclohexyl.
  • a "-" at the end of a group means that the group is attached to the rest of the molecule through that site.
  • halogen refers to fluorine, chlorine, bromine or iodine.
  • alkyl refers to a linear or branched, saturated, monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C 1 -C 6 ).
  • Alkyl includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, and the like.
  • alkoxy refers to the group R X -O-, where R X is defined as the term “alkyl”. Alkoxy includes, but is not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, and the like.
  • alkenylene is a divalent group connected to the rest of the molecule by two single bonds, and the rest of the definition is the same as the term “alkenyl”.
  • alkenyl refers to a linear or branched, unsaturated, monovalent hydrocarbon group with a specified number of carbon atoms (e.g., C2 - C6 ) and one or more (e.g., 1, 2, or 3) carbon-carbon sp2 double bonds.
  • Alkenyl includes, but is not limited to, vinyl, wait.
  • cycloalkyl refers to a cyclic, saturated, monovalent hydrocarbon radical having a specified number of carbon atoms (e.g., C 3 -C 6 ), which is a single ring. Cycloalkyl groups include, but are not limited to: wait.
  • aryl refers to a cyclic, unsaturated, monovalent hydrocarbon group with a specified number of carbon atoms (e.g., C 6 -C 10 ), which is a single ring or multiple rings (e.g., 2 or 3). When it is a multiple ring, the single rings share two atoms and one bond, and each ring has aromaticity.
  • Aryl includes, but is not limited to, phenyl, naphthyl, etc.
  • heterocycloalkyl refers to a cyclic, saturated, monovalent group having a specified number of ring atoms (e.g., 5-10 members), a specified number of heteroatoms (e.g., 1, 2, or 3), a specified heteroatom type (one or more of P, N, O, and S), which is monocyclic or polycyclic (e.g., 2 or 3 rings, bridged rings, spiro rings, or fused rings).
  • a bridged ring refers to a monocyclic ring that shares three or more carbon atoms and/or heteroatoms.
  • a fused ring refers to a monocyclic ring that shares one carbon atom.
  • a fused ring refers to a monocyclic ring that shares two carbon atoms and/or heteroatoms.
  • a heterocycloalkyl group is connected to the rest of the molecule via a carbon atom or a heteroatom. Heterocycloalkyl groups include, but are not limited to: wait.
  • heteroaryl refers to a cyclic, unsaturated, monovalent group having a specified number of ring atoms (e.g., 5-10 members), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (one or more of P, N, O, and S). It is a single ring or multiple rings (for example, 2 or 3), the single rings share two atoms and one bond, and each ring is aromatic.
  • the heteroaryl group is connected to the rest of the molecule through a carbon atom or a heteroatom; the heteroaryl group is connected to the rest of the molecule through a ring with a heteroatom or a ring without a heteroatom.
  • Heteroaryl groups include, but are not limited to: wait.
  • heteroaryl satisfies at least one of the following conditions, and the rest of the definition is the same as the term “heteroaryl”: 1. It is connected to the rest of the molecule through two or more single bonds; 2. It shares two atoms and one bond with the rest of the molecule.
  • isotope label refers to an isotope-labeled compound in which one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number normally occurring in nature, compared to the nitrogen-containing compound shown in Formula I.
  • isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F, and Cl, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 32 P, 35 S, 18 F, and 36 Cl, respectively.
  • Compounds of the present invention containing the above-mentioned isotopes and/or other isotopes of other atoms, pharmaceutically acceptable salts thereof, solvates thereof, solvates of pharmaceutically acceptable salts thereof, or prodrugs thereof are within the scope of the present invention.
  • Certain isotope-labeled compounds of the present invention for example compounds incorporating radioactive isotopes (such as 3 H and 14 C) can be used for drug and/or substrate tissue distribution assays. Tritium (i.e., 3 H) and carbon 14 (i.e., 14 C) isotopes are particularly preferred due to ease of preparation and detectability.
  • substitution with heavier isotopes may provide certain therapeutic advantages (e.g., increased in vivo half-life or reduced dosage requirements) derived from greater metabolic stability, and therefore may be preferred in certain circumstances.
  • the compounds of the invention as claimed in the claims may be specifically limited to substitution with deuterium or tritium.
  • the presence of hydrogen in a substituent without the term deuterium or tritium being separately listed does not exclude deuterium or tritium, but may also contain deuterium or tritium.
  • therapeutically effective amount refers to an amount administered to a patient that is sufficient to effectively treat a disease.
  • the therapeutically effective amount will vary depending on the type of compound, the type of disease, the severity of the disease, the age of the patient, etc., but can be adjusted by those skilled in the art as appropriate.
  • pharmaceutical excipients refers to all substances contained in pharmaceutical preparations other than active pharmaceutical ingredients, which are generally divided into two categories: excipients and additives. For details, please refer to the “Pharmacopoeia of the People's Republic of China (2020 Edition)” and Handbook of Pharmaceutical Excipients (Paul J Sheskey, Bruno C Hancock, Gary P Moss, David J Goldfarb, 2020, 9th Edition).
  • treating refers to removing the cause or alleviating the symptoms.
  • prevention refers to reducing the risk of developing a disease.
  • patient refers to any animal, usually a mammal, such as a human, that needs to be treated or prevented. Mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc.
  • the reagents and raw materials used in the present invention are all commercially available, among which It is commercially available and its CAS number is 15761-39-4.
  • the positive improvement effect of the present invention is that the compounds of the present invention have good inhibitory activity on DNA polymerase ⁇ .
  • FIG1 is an ellipsoid diagram of the molecular stereostructure of compound I-60 under single crystal diffraction.
  • FIG2 is an ellipsoid diagram of the molecular stereostructure of compound I-64 under single crystal diffraction.
  • the single crystal structure was solved by the direct method using SHELXT2014, and the structure was refined using the least squares method.
  • the hydrogen atom refinement process was obtained using isotropic calculations.
  • the hydrogen atoms on N and O were obtained through residual electron density, and the hydrogen atoms on C-H were obtained through calculated hydrogenation, and the riding model was used to refine them.
  • Step 2 tert-Butyl (S)-2-((2-methoxy-5-methylphenyl)carbamoyl)pyrrolidine-1-carboxylate
  • N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (3.7 g, 11.8 mmol) and 1-methylimidazole (0.53 mL, 6.69 mmol) were added to a solution of 2-methoxy-5-methylaniline (1.2 g, 5.58 mmol) and (tert-butyloxycarbonyl)-L-proline (764 mg, 5.58 mmol) in acetonitrile (20 mL).
  • the reaction solution was stirred at room temperature for 1 hour. Water (10 mL) was added and extracted with ethyl acetate (30 mL x 3).
  • Step 3 tert-Butyl (S)-2-((2-methoxy-5-methylphenyl)(methyl)carbamoyl)pyrrolidine-1-carboxylate
  • Step 5 (S)-1-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-N-(2-methoxy-5-methylphenyl)-N-methylpyrrolidine-2-carboxamide
  • N,N-diisopropylethylamine (936 mg, 7.25 mmol) was added to a solution of (S)-N-(2-methoxy-5-methylphenyl)-N-methylpyrrolidine-2-carboxamide (900 mg, 3.62 mmol) and 2,6-dichloro-4-(trifluoromethyl)pyridine (783 mg, 3.62 mmol) in 1,4-dioxane (10 mL).
  • the reaction solution was heated to 90°C and stirred for 2 hours, then cooled to room temperature. Water (30 mL) was added and extracted with ethyl acetate (30 mL x 3).
  • Step 6 (S,E)-1-(6-(2-ethoxyvinyl)-4-(trifluoromethyl)pyridin-2-yl)-N-(2-methoxy-5-methylphenyl)-N-methylpyrrolidine-2-carboxamide
  • Step 7 (S)-N-(2-hydroxy-5-methylphenyl)-N-methyl-1-(6-(2-carbonylethyl)-4-(trifluoromethyl)pyridin-2-yl)pyrrolidine-2-carboxamide
  • Step 8 (S)-N-(2-hydroxy-5-methylphenyl)-1-(6-(2-hydroxyethyl)-4-(trifluoromethyl)pyridin-2-yl)-N-methylpyrrolidine-2-carboxamide
  • Step 9 (S)-5 5 ,4-dimethyl-1 4 -(trifluoromethyl)-6-oxa-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctan-3-one (I-1)
  • Triphenylphosphine (123 mg, 0.47 mmol) and diisopropyl azodicarboxylate (95 mg, 0.47 mmol) were added to a tetrahydrofuran solution of (S)-N-(2-hydroxy-5-methylphenyl)-1-(6-(2-hydroxyethyl)-4-(trifluoromethyl)pyridin-2-yl)-N-methylpyrrolidine-2-carboxamide (200 mg, 50% purity, 0.24 mmol) at 0°C, and the mixture was heated to room temperature and the reaction was continued for 3 hours. After the reaction was completed, water (20 mL) was added and the mixture was extracted with ethyl acetate (30 mL x 3).
  • Step 3 tert-Butyl (S)-2-((2-allyl-5-methylphenyl)carbamoyl)pyrrolidine-1-carboxylate
  • Step 4 tert-Butyl (S)-2-((2-allyl-5-methylphenyl)(methyl)carbamoyl)pyrrolidine-1-carboxylate
  • Step 6 (S)-N-(2-allyl-5-methylphenyl)-1-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-N-methylpyrrolidine-2-carboxamide
  • Step 7 (S)-N-(2-allyl-5-methylphenyl)-N-methyl-1-(4-(trifluoromethyl)-6-vinylpyridin-2-yl)pyrrolidine-2-carboxamide
  • reaction solution was replaced with argon three times, heated to 80°C and stirred for 4 hours under an argon atmosphere. After the reaction was completed, the reaction solution was cooled to room temperature, filtered, and the filtrate was extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 8 (S,Z)-5 5 ,4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-7-ene-3-one (I-5)
  • Step 9 (S)-5 5 ,4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctan-3-one (I-4)
  • Step 10 (S)-5 5 ,4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 ,3-dione (I-12)
  • Step 1 tert-Butyl (6-chloro-4-(trifluoromethyl)pyridin-2-yl)carbamate
  • Step 2 tert-Butyl (E)-(6-(2-ethoxyvinyl)-4-(trifluoromethyl)pyridin-2-yl)carbamate
  • tert-butyl (6-chloro-4-(trifluoromethyl)pyridin-2-yl)carbamate (900 mg, 3.03 mmol) was dissolved in acetonitrile. (10mL), then add (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (1.21g, 6.11mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (222mg, 0.3mmol) and potassium carbonate (836mg, 6.06mmol). The reaction solution was purged with nitrogen three times, and then stirred at 80°C for 1 hour under nitrogen protection.
  • Step 4 tert-Butyl (E)-(6-(2-ethoxyvinyl)-4-(trifluoromethyl)pyridin-2-yl)(2-((2-methoxy-5-methylphenyl)(methyl)amino)-2-oxoethyl)carbamate
  • Step 5 N-(2-hydroxy-5-methylphenyl)-N-methyl-2-((6-(2-oxoethyl)-4-(trifluoromethyl)pyridin-2-yl)amino)acetamide
  • Step 6 N-(2-hydroxy-5-methylphenyl)-2-((6-(2-hydroxyethyl)-4-(trifluoromethyl)pyridin-2-yl)amino)-N-methylacetamide
  • lithium borohydride (0.2 mL, 2 mol/L, 0.39 mmol) tetrahydrofuran solution was slowly added dropwise to a tetrahydrofuran (5 mL) solution of N-(2-hydroxy-5-methylphenyl)-N-methyl-2-((6-(2-oxoethyl)-4-(trifluoromethyl)pyridin-2-yl)amino)acetamide (150 mg, 0.39 mmol), and then the temperature was raised to 20 ° C and stirred for 0.5 h.
  • Step 7 5-4,6 -dimethyl- 1-4- (trifluoromethyl)-4-oxa-6,9-diaza-1(2,6)-pyridine-5(1,2)-benzocyclononane-7-one (I-8)
  • Step 1 tert-Butyl (S)-2-((2-bromo-5-methylphenyl)carbamoyl)-2,5-dihydro-1H-pyrrole-1-carboxylate
  • Step 2 tert-Butyl (2S,3S,4R)-2-((2-bromo-5-methylphenyl)carbamoyl)-3,4-dihydroxypyrrolidine-1-carboxylate
  • Step 3 tert-Butyl (3aS, 4S, 6aR)-4-((2-bromo-5-methylphenyl)carbamoyl)-2,2-dimethyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-5-carboxylate
  • Step 4 tert-Butyl (3aS, 4S, 6aR)-4-((2-bromo-5-methylphenyl)(methyl)carbamoyl)-2,2-dimethyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-5-carboxylate
  • Step 5 tert-Butyl (3aS, 4S, 6aR)-4-((2-allyl-5-methylphenyl)(methyl)carbamoyl)-2,2-dimethyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-5-carboxylate
  • Step 6 (3aS,4S,6aR)-N-(2-allyl-5-methylphenyl)-N,2,2-trimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • Step 7 (3aS, 4S, 6aR)-N-(2-allyl-5-methylphenyl)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-N,2,2-trimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • N,N-diisopropylethylamine (782 mg, 6.05 mmol) was added to a DMF (10 mL) solution of (3aS,4S,6aR)-N-(2-allyl-5-methylphenyl)-N,2,2-trimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide (500 mg, 1.51 mmol) and 2,6-dichloro-4-(trifluoromethyl)pyridine (654 mg, 3.03 mmol), and the mixture was stirred at 120°C for 2 hours.
  • Step 8 (3aS,4S,6aR)-N-(2-allyl-5-methylphenyl)-N,2,2-trimethyl-5-(4-(trifluoromethyl)-6-vinylpyridin-2-yl)tetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • Step 9 (2 3a R, 2 6 S, 2 6a S, Z)-2 2 , 2 2 , 5 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctane-7-en-3-one
  • Step 10 (2 3a R, 2 6 S, 2 6a S, Z)-2 2 , 2 2 , 5 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-phenylcyclooctan-3-one
  • Step 11 (2 3a S, 2 6 S, 2 6a S)-2 2 , 2 2 , 5 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-phenylcyclooctane-2 4 ,3-dione
  • reaction solution was diluted with water (10 mL), the aqueous phase was extracted twice with ethyl acetate (20 mL), the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 12 (2 2 S, 2 3 S, 2 4 S )-2 3 , 2 4 -dihydroxy- 5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 , 3-dione (I-14)
  • Step 7 (3aS, 4S, 6aR)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid methyl ester
  • (2S)-1-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-3,4-dihydroxypyrrolidine-2-carboxylic acid methyl ester (4.3 g, 12.6 mmol) was dissolved in acetone (30 mL), and 2,2-dimethoxypropane (65.7 g, 631 mmol) and PPTS (3.81 g, 15.2 mmol) were added to the reaction solution.
  • reaction solution was heated to 60°C and stirred for 3 hours.
  • the reaction solution was cooled to room temperature, water (30 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 8 (3aS, 4S, 6aR)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid
  • Step 9 (3aS, 4S, 6aR)-N-(5-allyl-1-methyl-1H-indazol-6-yl)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • Step 10 (3aS, 4S, 6aR)-N-(5-allyl-1-methyl-1H-indazol-6-yl)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-N,2,2-trimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • reaction solution was stirred at 0°C for 0.5 hours, and then iodomethane (0.15 mL, 2.46 mmol) was added to the solution.
  • the reaction solution was warmed to room temperature and stirred for 1 hour.
  • Add The reaction was quenched with saturated aqueous ammonium chloride solution (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 11 (3aS,4S,6aR)-N-(5-allyl-1-methyl-1H-indazol-6-yl)-N,2,2-trimethyl-5-(4-(trifluoromethyl)-6-vinylpyridin-2-yl)tetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • reaction solution was replaced with argon three times, and heated to 80°C under argon atmosphere and stirred for 8 hours. After the reaction was completed, the reaction solution was cooled to room temperature, filtered, and water (20 mL) and ethyl acetate (30 mL) were added to the filtrate. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 12 (2 3a R, 2 6 S, 2 6a S, Z)-2 2 , 2 2 , 5 1 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H, 5 1 H-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-5(6,5)-indazole-1(2,6)-pyridinecyclooctane-7-ene-3-one
  • Step 13 (2 3a R, 2 6 S, 2 6a S)-2 2 , 2 2 , 5 1 , 4-tetramethyl- 1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H, 5 1 H-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-5(6,5)-indazole-1(2,6)-pyridinecyclooctan-3-one
  • Step 14 (2 3a S, 2 6 S, 2 6a S)-2 2 , 2 2 , 5 1 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H, 5 1 H-4-aza-2(5,6)- [1,3]dioxazolo[4,5-c]pyrrole-5(6,5)-indazole-1(2,6)-pyridinecyclooctane-2 4 ,3-dione
  • Step 15 (2 2 S, 2 3 S, 2 4 S )-2 3 , 2 4 -dihydroxy-5 1 , 4-dimethyl-1 4 -(trifluoromethyl)-5 1 H-4-aza-5(6,5)-indazole-1(2,6)-pyridine-2(1,2)-pyrrolecyclooctane-2 5 ,3-dione (I-24)
  • Imidazole (5.29 g, 77.7 mmol) was dissolved in dichloromethane (30 mL), and triphenylphosphine (8.15 g, 31.1 mmol) was added. The reaction solution was stirred at room temperature for 1 hour. Iodine (7.89 g, 31.1 mmol) was added in batches. The mixture was stirred at room temperature for 1 hour. 4-Methyl-2-nitrobenzyl alcohol (4.33 g, 25.9 mmol) was added in batches. The mixture was stirred at room temperature for 1 hour.
  • Step 3 (3aS,4S,6aR)-5-(6-hydroxymethyl-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid methyl ester
  • Step 4 (3aS,4S,6aR)-2,2-dimethyl-5-(6-(((4-methyl-2-nitrobenzyl-)oxy)methyl)-4-(trifluoromethyl)pyridin-2-yl)-tetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid methyl ester
  • Step 5 (3aS,4S,6aR)-5-(6-(((2-amino-4-methylbenzyl)oxy)methyl)-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid methyl ester
  • Step 6 (3aS,4S,6aR)-5-(6-(((2-amino-4-methylbenzyl)oxy)methyl)-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid
  • Step 7 (2 3a R, 2 6 S, 2 6a S)-2 2 , 2 2 , 2 5 -trimethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-7-oxa-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole 1(2,6)-pyridine-5(1,2)-benzocyclooctan-3-one
  • Step 8 (2 3a R, 2 6 S, 2 6a S)-2 2 , 2 2 , 2 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-7-oxa-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole 1(2,6)-pyridine-5(1,2)-benzocyclooctan-3-one
  • Step 9 (2 3a S, 2 6 S, 2 6a S)-2 2 , 2 2 , 2 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-7-oxa-4-aza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctane-2 4 ,3-dione
  • reaction solution was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was filtered and extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated.
  • Step 10 (2 2 S, 2 3 S, 2 4 S)-2 3 , 2 4 -dihydroxy- 5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-7-oxa-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 ,3-dione (I-33)
  • Step 3 (2S,3R)-2-((2-allyl-5-methylphenyl)carbamoyl)-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester
  • Step 4 (tert-Butyl (2S, 3R)-2-((2-allyl-5-methylphenyl)carbamoyl)-3-((tert-butyldimethylsilyl)oxy)pyrrolidine-1-carboxylate
  • Step 5 tert-Butyl (2S, 3R)-2-((2-allyl-5-methylphenyl)(methyl)carbamoyl)-3-((tert-butyldimethylsilyl)oxy)pyrrolidine-1-carboxylate
  • Step 7 (2S,3R)-N-(2-allyl-5-methylphenyl)-3-((tert-butyldimethylsilyl)oxy)-1-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-N-methylpyrrolidine-2-carboxamide
  • Step 8 (2S,3R)-N-(2-allyl-5-methylphenyl)-3-((tert-butyldimethylsilyl)oxy)-N-methyl-1-(4-(trifluoromethyl)-6-vinylpyridin-2-yl)pyrrolidine-2-carboxamide
  • Step 9 (2 2 S, 2 3 R, Z)-2 3 -((tert-butyldimethylsilyl)oxy)-5 5 ,4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-7-ene-3-one
  • Step 10 (2 2 S, 2 3 R)-2 3 -((tert-butyldimethylsilyl)oxy)-5 5 ,4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctan-3-one
  • Step 11 (2 2 S, 2 3 R)-2 3 -hydroxy-5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctan-3-one (I-36)
  • Step 1 (2 2 S, 2 3 R)-2 3 -((tert-butyldimethylsilyl)oxy)-5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane- 2 5 , 3-dione
  • Step 2 (2 2 S, 2 3 R)-2 3 -hydroxy-5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 ,3-dione (I-34)
  • Step 1 tert-Butyl (2S,3R,4S)-2-((2-bromo-5-methylphenyl)carbamoyl)-3,4-dihydroxypyrrolidine-1-carboxylate
  • Step 2 tert-Butyl (3aR, 4S, 6aS)-4-((2-bromo-5-methylphenyl)carbamoyl)-2,2-dimethyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-5-carboxylate
  • Step 3 tert-Butyl (3aR, 4S, 6aS)-4-((2-bromo-5-methylphenyl)(methyl)carbamoyl)-2,2-dimethyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-5-carboxylate
  • Step 4 tert-Butyl (3aR, 4S, 6aS)-4-((2-allyl-5-methylphenyl)(methyl)carbamoyl)-2,2-dimethyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-5-carboxylate
  • Step 6 (3aR, 4S, 6aS)-N-(2-allyl-4-methylphenyl)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-N,2,2-trimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • Step 7 (3aR, 4S, 6aS)-N-(2-allyl-4-methylphenyl)-N,2,2-trimethyl-5-(4-(trifluoromethyl)-6-vinylpyridin-2-yl)tetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • Step 8 (2 3a R, 2 4 S, 2 6a S, Z)-2 2 , 2 2 , 5 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4-aza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-phenylcyclooctane-7-ene-3-one
  • Step 9 (2 3a R, 2 4 S, 2 6a S)-2 2 , 2 2 , 5 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4-aza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctan-3-one
  • Step 10 (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 5 5 , 4-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4-aza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-phenylcyclooctane-2 6 ,3-dione
  • reaction solution was diluted with water (50 mL), the aqueous phase was extracted with ethyl acetate (50 mL x 2), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 11 (2 2 S, 2 3 R, 2 4 R )-2 3 , 2 4 -dihydroxy-5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-4-aza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 , 3-dione (I-38)
  • Step 3 (2S)-1-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-3,4-dihydroxypyrrolidine-2-carboxylic acid methyl ester
  • Step 4 (3aR, 4S, 6aS)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid methyl ester
  • (2S)-1-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-3,4-dihydroxypyrrolidine-2-carboxylic acid methyl ester (4.3 g, 12.6 mmol) was dissolved in acetone (30 mL), and 2,2-dimethoxypropane (65.7 g, 631 mmol) and pyridine p-toluenesulfonate (3.81 g, 15.2 mmol) were added to the reaction solution.
  • reaction solution was heated to 60°C and stirred for 3 hours.
  • the reaction solution was cooled to room temperature, water (30 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 5 (3aR, 4S, 6aS)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid
  • Step 6 (3aR, 4S, 6aS)-5-(6-chloro-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid benzyl ester
  • Step 7 (3aR, 4S, 6aS)-5-(6-((tert-butoxycarbonyl)amino)-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid benzyl ester
  • Step 8 (3aR, 4S, 6aR)-5-(6-((tert-butoxycarbonyl)amino)-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid benzyl ester
  • Step 9 (3aR, 4S, 6aR)-5-(6-amino-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid benzyl ester
  • Step 10 (3aR, 4S, 6aR)-2,2-dimethyl-5-(6-((4-methyl-2-nitrophenylethyl)amino)-4-(trifluoromethyl)pyridin-2-yl)-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid benzyl ester
  • Step 11 (3aR, 4S, 6aR)-5-(6-((2-amino-4-methylphenethyl)amino)-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid
  • Step 12 (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 5-trimethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctane-2 6 ,3-dione
  • Step 13 (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 5 5 , 4,8-pentamethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctanone-2 6 ,3-dione
  • reaction solution was warmed to room temperature and stirred for 2 hours.
  • Saturated aqueous ammonium chloride solution was added to the solution to quench the reaction, and the mixture was extracted with ethyl acetate (30 mL x 3).
  • the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 14 (2 2 S, 2 3 R, 2 4 R)-2 3 , 2 4 -dihydroxy- 5 5 , 4 , 8-trimethyl-1 4 -(trifluoromethyl)-4, 8-diaza-1(2, 6)-pyridine-2(1, 2)-pyrrolidine-5(1, 2)-benzocyclooctane- 2 5 , 3-dione
  • Step 1 (3aR,4S,6aR)-5-(6-((tert-butoxycarbonyl)(4-methyl-2-nitrophenylethyl)amino)-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid benzyl ester
  • Step 2 (3aR, 4S, 6aR)-5-(6-((2-amino-4-methylphenethyl)(tert-butoxycarbonyl)amino)-4-(trifluoromethyl)pyridin-2-yl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid
  • Step 3 tert-Butyl (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 5 5 -trimethyl-2 6 , 3-dicarbonyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctane-8-carboxylate
  • Step 4 tert-Butyl (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 5 5 -trimethyl-4-(methyl-d3)-2 6 , 3-dicarbonyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctane-8-carboxylate
  • Step 5 (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 5 5 -trimethyl-4-(methyl-d3)-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctanone-2 6 ,3-dione
  • Step 6 (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 5 5 -8-tetramethyl-4-(methyl-d3)-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctanone-2 6 ,3-dione
  • Step 7 (2 2 S, 2 3 R, 2 4 R )-2 3 , 2 4 -dihydroxy- 5 5 , 8-dimethyl-4-(methyl-d 3 )-1 4 -(trifluoromethyl)-4,8-diaza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 , 3-dione
  • the Flack constant was: 0.07(13), C18 and C19 were R configuration, and C20 was S configuration.
  • step 4 is replaced by iodomethane instead of deuterated iodomethane
  • step 6 is replaced by deuterated iodomethane instead of original iodomethane
  • the Flack constant was 0.05(8), and the chirality of C3, C4 and C17 was S configuration.
  • step thirteen The same synthetic route is used as compound I-59, and the starting material of step thirteen is replaced by deuterated iodomethane instead of the original iodomethane.
  • step ten The same synthetic route was used as compound I-59, and the starting material of step ten was replaced by 2-(4-(methyl-d3)-2-nitrophenyl)acetaldehyde.
  • Step 1 tert-Butyl N-(6-bromo-4-(trifluoromethyl)pyridin-2-yl)carbamate
  • tert-butyl N-(6-bromo-4-(trifluoromethyl)pyridin-2-yl)carbamate (7.0 g, 20.5 mmol) was dissolved in dichloromethane. (10mL) solution, and trifluoroacetic acid (5mL) was added to the reaction solution, and the reaction solution was stirred at room temperature for 30 minutes. After the reaction was completed, a saturated sodium bicarbonate aqueous solution (100mL) was poured into the reaction solution, and extracted with ethyl acetate (50mL x 2).
  • 6-bromo-4-(trifluoromethyl)pyridine-2-amine 4.7g, 19.5mmol
  • 2-(4-methyl-2-nitro-phenyl)acetaldehyde 4.2g, 23.4mmol
  • acetic acid 5mL
  • sodium cyanoborohydride 1.6g, 25.4mmol
  • Step 4 6-Bromo-N-methyl-N-(4-methyl-2-nitrophenylethyl)-4-(trifluoromethyl)pyridin-2-amine
  • 6-bromo-N-(2-4-methyl-2-nitro-phenyl)ethyl)-4-(trifluoromethyl)pyridine-2-amine (3.7 g, 9.1 mmol) was dissolved in N,N-dimethylformamide (30 mL). After the reaction solution was cooled to 0°C, sodium hydride (0.26 g, 10.9 mmol) was added to the reaction solution. After the reaction solution was stirred at 0°C for 10 minutes, iodomethane (0.56 mL, 9.1 mmol) was added to the reaction solution, and the reaction solution was continued to stir at 0°C for 30 minutes.
  • Step 5 N-(2-amino-4-methylphenethyl)-6-bromo-N-methyl-4-(trifluoromethyl)pyridin-2-amine
  • 6-bromo-N-methyl-N-(4-methyl-2-nitrophenylethyl)-4-(trifluoromethyl)pyridine-2-amine (3.66 g, 8.75 mmol) was dissolved in ethanol (40 mL), and iron powder (2.44 g, 43.8 mmol), ammonium chloride (2.34 g, 43.8 mmol) and water (10 mL) were added to the reaction solution.
  • the reaction solution was stirred at 60° C. for 1 hour. After the reaction was completed, the reaction solution was cooled to room temperature and ethyl acetate (200 mL) was added to the reaction solution.
  • Step 6 (3aS, 4S, 6aS)-N-(2-(2-((6-bromo-4-(trifluoromethyl)pyridin-2-yl)(methyl)amino)ethyl)-5-methylphenyl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • N-(2-amino-4-methylphenethyl)-6-bromo-N-methyl-4-(trifluoromethyl)pyridin-2-amine (3.28 g, 8.45 mmol)
  • (3aS, 4S, 6aS)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid (1.7 g, 8.45 mmol)
  • 1-methylimidazole (2.02 mL, 25.4 mmol) were dissolved in N,N-dimethylformamide (30 mL), and the reaction solution was cooled to 0°C.
  • Tetramethyl chlorouronium hexafluorophosphate (2.84 g, 10.1 mmol) was dissolved in N,N-dimethylformamide (10 mL) and added dropwise to the reaction solution, and the reaction solution was stirred at 0°C for 2 hours. After the reaction was completed, the reaction solution was poured into water and the two were extracted with ethyl acetate (30 mL). The organic phases were combined and washed twice with half-saturated brine (100 mL).
  • Step 7 (2 3a S, 2 6 S, 2 6a S)-2 2 , 2 2 , 5 5 , 8-tetramethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4,8-diaza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctane- 2 4 ,3-dione (3aS,4S,6aS)-N-(2-(2-((6-bromo-4-(trifluoromethyl)pyridin-2-yl)(methyl)amino)ethyl)-5-methylphenyl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide (2.35 g, 4.11 mmol) was dissolved in
  • reaction solution was stirred at 100° C. for 6 hours. After the reaction was completed, the reaction solution was poured into water, and aqueous ammonia (2 mL) was added to the water. After sufficient stirring, the reaction solution was extracted twice with ethyl acetate (100 mL). The organic phases were combined and washed twice with semi-saturated brine (200 mL).
  • Step 8 (2 3a S, 2 6 S, 2 6a S)-2 2 , 2 2 , 5 5 , 8-tetramethyl-4-(methyl-d3)-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-4,8-diaza-2(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-1(2,6)-pyridine-5(1,2)-benzocyclooctane- 2 4 ,3-dione
  • reaction solution was cooled to 0° C.
  • sodium hydride (0.07 g, 2.8 mmol, 60%) was added to the reaction solution, and the reaction solution was stirred at 0° C. for 10 minutes.
  • Deuterated iodomethane (0.34 g, 2.34 mmol) was added to the reaction solution, and the reaction solution was stirred for 30 minutes at 0° C.
  • the reaction solution was poured into a saturated aqueous ammonium chloride solution (100 mL) and extracted twice with ethyl acetate (50 mL). The organic phases were combined and washed twice with semi-saturated brine (100 mL).
  • Step 9 (2 2 S, 2 3 S, 2 4 S )-2 3 , 2 4 -dihydroxy- 5 5 , 8-dimethyl-4-(methyl-d3)-1 4 -(trifluoromethyl)-4,8-diaza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 , 3-dione
  • step 4 The same synthetic route was used as for compound I-64, except that the starting material of step 4 was replaced by deuterated iodomethane instead of the original iodomethane.
  • Step 3 (3aR,4S,6aS)-2,2,6a-trimethyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-4,5-dicarboxylic acid di-tert-butyl ester
  • Step 4 (3aR, 4S, 6aS)-2,2,6a-methyl-6-carbonyltetrahydro-5H-[1,3]dioxazolo[4,5-c]pyrrole-4,5-dicarboxylic acid di-tert-butyl ester
  • Step 5 (3aR, 4S, 6aS)-2,2,6a-trimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxylic acid
  • Step 6 (3aR, 4S, 6aR)-N-(2-(2-((6-bromo-4-(trifluoromethyl)pyridin-2-yl)(methyl)amino)ethyl)-5-methylphenyl)-2,2,6a-trimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • Step 7 (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 2 6a , 5 5 , 8-pentamethyl-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro-2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrolidine-1(2,6)-pyridine-5(1,2)-benzocyclooctane- 2 6 ,3-dione
  • Step 8 (2 3a R, 2 4 S, 2 6a R)-2 2 , 2 2 , 2 6a , 5 5 , 8-pentamethyl-4-(methyl-d 3 )-1 4 -(trifluoromethyl)-2 3a , 2 5 , 2 6 , 2 6a -tetrahydro- 2 4 H-4,8-diaza-2(5,4)-[1,3]dioxazolo[4,5-c]pyrrolidine-1(2,6)-pyridine-5(1,2)-benzocyclooctane- 2 6 ,3-dione
  • Step 9 (2 2 S, 2 3 R, 2 4 R)-2 3 , 2 4 -dihydroxy-2 4 , 5 5 , 8-trimethyl-4-(methyl-d 3 )-1 4 -(trifluoromethyl)-4,8-diaza-1(2,6)-pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocyclooctane-2 5 , 3-dione
  • Step 5 (6-bromo-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)(phenyl)methanone
  • Step 7 6-Bromo-1-(4-methyl-2-nitrophenylethyl)-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine
  • 6-Bromo-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine (0.6 g, 2.26 mmol) was dissolved in tetrahydrofuran (10 mL), triphenylphosphine (1.18 g, 4.53 mmol), 2-(4-methyl-2-nitrophenyl)ethane-1-ol (0.61 g, 3.4 mmol) and diisopropyl azodicarboxylate (0.96 g, 4.75 mmol) were added to the reaction solution, and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, water (20 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL x 3).
  • Step 8 2-(2-(6-bromo-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl)-5-methylaniline
  • 6-Bromo-1-(4-methyl-2-nitrophenylethyl)-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridine (0.7 g, 1.63 mmol) was dissolved in ethanol (6 mL) and water (2 mL), iron powder (0.46 g, 8.17 mmol) and ammonium chloride (0.44 g, 8.17 mmol) were added to the reaction solution, and the mixture was stirred at 80°C for 1 hour. After the reaction was completed, the mixture was filtered and the filtrate was extracted with ethyl acetate (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue.
  • Step 9 (3aS, 4S, 6aS)-N-(2-(2-(6-bromo-4-(trifluoromethyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)ethyl)-5-methylphenyl)-2,2-dimethyl-6-carbonyltetrahydro-4H-[1,3]dioxazolo[4,5-c]pyrrole-4-carboxamide
  • Step 10 (1 3a S, 1 6 S, 1 6a S)-1 2 , 1 2 , 5 4 -trimethyl- 2 4 - (trifluoromethyl)-1 3a , 1 5 , 1 6 , 1 6a -tetrahydro- 1 4 H, 2 1 H-6-aza-1(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-2(6,1)-pyrrolo[2,3-b]pyridine-5(1,2)-benzocycloheptane- 1 4 , 7-dione
  • Step 11 (1 3 a S, 1 6 S, 1 6 a S)-1 2 , 1 2 , 5 4 , 6-tetramethyl- 2 4 -(trifluoromethyl)-1 3 a , 1 5 , 1 6 , 1 6 a -tetrahydro-1 4 H, 2 1 H-6-aza-1(5,6)-[1,3]dioxazolo[4,5-c]pyrrole-2(6,1)-pyrrolo[2,3-b]pyridine-5(1,2)-benzocycloheptane-1 4 , 7-dione
  • Step 12 (2 2 S, 2 3 S, 2 4 S )-2 3 , 2 4 -dihydroxy-5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-1 1 H-4-aza-1(6,1)-pyrrolo[2,3-b]pyridine-2(1,2)-pyrrolidine-5(1,2)benzocycloheptane- 2 5 , 3-dione
  • Step 13 (2 2 S, 2 3 S, 2 4 S)-2 3 , 2 4 -dihydroxy-5 5 , 4-dimethyl-1 4 -(trifluoromethyl)-1 2 , 1 3 -dihydro-1 1 H-4-aza-1(6,1)-pyrrolo[2,3-b]pyridine-2(1,2)-pyrrolidine-5(1,2)-benzocycloheptane-2 5 , 3-dione
  • the inhibitory effect of the compounds on Pol ⁇ polymerase activity was evaluated by PicoGreen assay.
  • the compound was dissolved in 100% DMSO, and added to a 384-well plate (corning Cat#4514) after serial dilution.
  • the initial (final) concentration was 12 ⁇ M, and the dilution was 4-fold, with a total of 10 concentration points.
  • purified recombinant Pol ⁇ protein (aa1792-2590) and a 1:1.1 ratio of primer ( 5' -GCG GCT GTC ATA AG-3'): template (5'-GCT ACA TTG ACA ATG GCA TCA AAT CTC AGA TTG CGT CTT ATG ACA GCC GCG-3') duplex were added in an assay reaction buffer containing 25 mM Tris (pH 7.5), 12.5 mM NaCl, 0.5 mM MgCl2, 1 mM DTT, 0.01% (v/v) Triton X-100, 0.01% (v/v) BSA, and 5% glycerol to a final concentration of 1.5 nM, 50 nM, and 55 nM, respectively.
  • primer 5' -GCG GCT GTC ATA AG-3'
  • template 5'-GCT ACA TTG ACA ATG GCA TCA AAT CTC AGA TTG CGT CTT ATG ACA GCC GCG
  • dNTPs were added at a final concentration of 40 ⁇ M.
  • the DMSO concentration in each well was maintained at 1%, and incubated at room temperature for 60 minutes.
  • Use ENVISION to read the fluorescence and analyze it under the 485/520nm module.
  • the RFU value normalized by the control well value in each assay plate was used to calculate the inhibition percentage (%) of each concentration of the compound, and XLfit was used for curve fitting. The specific results are shown in Table 1.
  • Ref.1 is Selected from WO2021028670A1.
  • TAEJ ta-mediated end joining
  • HEK293T cells were cultured in DMEM supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin-streptomycin (PS).
  • FBS heat-inactivated fetal bovine serum
  • PS penicillin-streptomycin
  • the cell culture plate was equilibrated at room temperature for 10 min.
  • TBEJ Theta-mediated end-joining
  • DLD-1 BRCA2 -/- and DLD-1 BRCA2 WT cell proliferation assay
  • the CellTiter-Glo (CTG) method was used to evaluate the antiproliferative effects of the compounds on human colorectal adenocarcinoma cell lines DLD-1 BRCA2 -/- and DLD-1 BRCA2 WT .
  • DLD-1 (purchased from ATCC, product number: CCL-221, batch number: 70033255)
  • Trypsine-EDTA purchased from Invitrogen, catalog number: 25200-072
  • Penicillin-Streptomycin solution (purchased from Hyclone, product number: SV30010)
  • FBS (purchased from BI, catalog number: 04-002-1A)
  • the antiproliferative effect of the compounds on human colorectal adenocarcinoma cell lines was evaluated by ATP quantification, and the activity correlated well with the number of viable cells.
  • the cell line DLD-1 (ATCC, CCL-221) and the genetically edited cell line DLD-1 BRCA2 -/- were cultured in RPMI 1640 (Invitrogen, 11875-085) medium supplemented with 10% (v/v) fetal bovine serum (BI, 04-002-1A).
  • the cell lines were cultured according to the standard instructions of the American Type Culture Collection. The cell lines were identified by short tandem repeat profiles.
  • cells were collected and resuspended in cell culture medium. Cell density was determined using a cell counter. An appropriate amount of cell suspension was removed from the centrifuge tube to prepare the seed plate working solution (100 cells/100 ⁇ L for DLD1 cell line and 150 cells/100 ⁇ L for DLD-1 BRCA2 -/- cell line). 100 ⁇ L of cell suspension was spread in each well of a 96-well plate (Corning, 3603) and incubated overnight in an incubator. The diluted test compound solution was added to the well plate. Fresh culture medium was replaced and the compound was re-treated on the fourth and seventh days of incubation of the compound, and the total incubation time was 10 days.
  • the cell culture plate was removed and equilibrated to room temperature. 100 ⁇ L CellTiter-Glo (Promega, G7558) was added to each well and incubated at room temperature for 30 minutes. The data on the plate was read using a microplate reader in 96-well Luminescence mode (Perkinelme, Enspire). The dose-response one-site 205 model of IDBS XLfit was used. Data analysis, curve fitting and reporting. See Table 3 for detailed results.
  • the positive control information and substrate information are shown in the following table:
  • liver microsomes were thawed in a 37°C water bath and the liver microsome working solution was prepared (the final concentration of the system was 0.5 mg/mL).
  • the above stock solution was prepared into 2 mM working solution with DMSO.
  • the inhibition percentage (% inhibition) was calculated using the metabolite formation of the test compound or control compound compared to the matrix control. Specific results are shown in Table 4.
  • N means no inhibition
  • Intravenous injection 40% HP- ⁇ -CD aqueous solution
  • intragastric administration 0.2% HPC + 0.5% Tween80 aqueous solution.
  • mice 6 male mice. Fasting treatment was performed in the evening of the day before administration, and water was freely available. During the experimental process, mice were free to eat and drink, and the drugs were administered by intravenous injection or gavage. After administration, the animal status was observed and abnormal manifestations were recorded. Blood was collected from the eye sockets at 0.0833, 0.25, 1, 2, 4, 8 and 24 hours after intravenous injection; blood was collected from the eye sockets at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after gavage.
  • the internal standard method was used to establish a standard curve, with the theoretical standard curve concentration as the horizontal axis and the peak area ratio as the vertical axis (test compound peak area/internal standard peak area).
  • the linear regression method (weighting factor 1/X2) was used, and R 2 >0.9900.
  • the unknown samples were calculated using the standard curve.
  • the non-compartmental analysis model of WinNonlin 8.2 software calculates the pharmacokinetic parameters and presents them in the report, including T 1/2 , T max , C max and AUC, etc. The specific results are shown in Table 5.
  • a PO is oral administration;
  • T 1/2 is the half-life of the drug;
  • c AUC 0-t is the area under the blood drug concentration-time curve from time 0 to the last selected time point;
  • d F% is the bioavailability.

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Abstract

La présente invention concerne un composé macrocyclique contenant de l'azote, son procédé de préparation et son utilisation. Le composé de la présente invention est tel que représenté par I. Le composé de la présente invention a une activité inhibitrice relativement bonne par rapport à l'ADN polymérase θ et a une bonne perspective pour être formé en un médicament.
PCT/CN2024/081244 2023-03-13 2024-03-12 Composé macrocyclique contenant de l'azote, son procédé de préparation et son utilisation Pending WO2024188246A1 (fr)

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CN114846003A (zh) * 2019-08-09 2022-08-02 阿提奥斯医药有限公司 用于治疗癌症的氘代化合物
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