WO2022228576A1 - 一种靶向蛋白调节剂的化合物及其应用 - Google Patents
一种靶向蛋白调节剂的化合物及其应用 Download PDFInfo
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- WO2022228576A1 WO2022228576A1 PCT/CN2022/090813 CN2022090813W WO2022228576A1 WO 2022228576 A1 WO2022228576 A1 WO 2022228576A1 CN 2022090813 W CN2022090813 W CN 2022090813W WO 2022228576 A1 WO2022228576 A1 WO 2022228576A1
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
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Definitions
- the present invention relates to a compound targeting protein regulator and its application.
- RAS represents a group of closely related monomeric globular proteins (21 kDa molecular weight) of 189 amino acids that associate with the plasma membrane and bind GDP or GTP. RAS acts as a molecular switch. When RAS contains bound GDP, it is in a resting or closed position and "inactive". In response to exposure of cells to certain growth-promoting stimuli, RAS is induced to exchange its bound GDP for GTP. In the case of binding GTP, RAS is "turned on” and is able to interact with and activate other proteins (its "downstream targets”). The RAS protein itself has a very low intrinsic ability to hydrolyze GTP back to GDP, thereby turning itself into the off state.
- GAP GTPase activating protein
- RAS proteins contain a G domain responsible for the enzymatic activity of RAS, guanine nucleotide binding and hydrolysis (GTPase reaction). It also contains a C-terminal extension called the CAAX box, which can be post-translationally modified and is responsible for targeting the protein to the membrane.
- the G domain is approximately 21-25 kDa in size and contains a phosphate-binding loop (P-loop).
- P-loop represents the pocket in the protein that binds nucleotides, and this is the rigid part of the domain with conserved amino acid residues necessary for nucleotide binding and hydrolysis (glycine 12, threo amino acid 26 and lysine 16).
- the G domain also contains the so-called switch I region (residues 30-40) and switch II region (residues 60-76), both of which are the dynamic part of the protein as it switches between rest and load states capability is often denoted as a "spring loaded” mechanism.
- the main interaction is the hydrogen bond formed by threonine-35 and glycine-60 with the ⁇ -phosphate of GTP, which allows the switch 1 and switch 2 domains, respectively, to maintain their active conformations. After hydrolysis of GTP and release of phosphate, both relax to the inactive GDP conformation.
- RAS The most notable members of the RAS subfamily are HRAS, KRAS and NRAS, which are primarily involved in many types of cancer.
- many other members exist including DIRAS1; DIRAS2; DIRAS3; ERAS; GEM; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS and RRAS2.
- KRAS Mutation in any of the three major isoforms of the RAS gene (HRAS, NRAS or KRAS) is one of the most common events in human tumorigenesis. About 30% of all human tumors were found to carry some mutation in the RAS gene. Notably, KRAS mutations are detected in 25%-30% of tumors. In contrast, the rates of oncogenic mutations in NRAS and HRAS family members were much lower (8% and 3%, respectively). The most common KRAS mutations are found in the P-loop at residues G12 and G13 and at residue Q61. Among tumor-associated KRAS G12 mutations, KRAS G12D has the highest mutation probability, about 40%.
- KRAS has been a target of interest for drug developers. Despite the progress made in this field, there is still a need in the art for improved KRAS G12D mutein inhibitors.
- PROTAC proteolysis-targeting chimeras
- PROTAC has two heterofunctional Compounds in which the ligands are linked via a linker: one ligand targets the protein of interest (POI), while the other specifically recruits the E3 ligase.
- POI protein of interest
- PROTAC binds E3 ligase and target protein, a ternary complex is formed.
- POI protein of interest
- PROTAC makes POI present a favorable steric position to promote its ubiquitination, thereby selectively reducing the level of target protein.
- the advantage of this method is that PROTAC can catalyze multiple rounds of degradation of target proteins, which is the biggest difference between PROTAC molecules and small molecule inhibitors.
- CN110684015A discloses a PROTAC molecule targeting ALK.
- the PROTAC molecule targeting ALK has been successfully prepared, which can effectively target the target protein, reduce the content of ALK in cells, and has good antitumor activity in vitro and in vivo. Low toxicity to normal cells, in line with the characteristics of high efficiency and low toxicity.
- no PROTAC molecules targeting KRAS G12D have been reported in the art.
- the technology to be solved in the present invention is to overcome the defect of the prior art that there are few types of compounds capable of targeting and degrading mutant KRAS.
- a PROTAC small molecule compound and its application are provided.
- the compounds of the present invention have a good inhibitory effect on the KRAS_G12D mutant.
- the present invention solves the above technical problems through the following technical solutions.
- the present invention provides a compound represented by formula Ia, Ib or Ic, its pharmaceutically acceptable salt, its solvate, its stereoisomer, its tautomer, its prodrug, its metabolism Product or its isotopic compound:
- R 1a , R 1b and R 1c are independently C 6 -C 18 aryl, C 6 -C 18 aryl substituted by R 1a-1 (R 1a-1 is one or more, such as 1, 2 or 3) or 5-10 heteroaryl groups, the heteroatoms in the 5-10 heteroaryl groups are independently one or more of N, O or S, and the number of heteroatoms is 1-4 (e.g. 1, 2, 3 or 4);
- R 1a-1 is independently hydroxy, halogen, C 1 -C 12 alkyl or C 2 -C 6 alkynyl;
- R 2a , R 4a , R 2b , R 4b , R 2c and R 5c are independently H or halogen;
- R 3a is a 7-12-membered bridged heterocycloalkyl, the heteroatoms in the bridged heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1 -4 (e.g. 1, 2, 3 or 4);
- R 4c is H, a 3-6 membered heterocycloalkyl or a 3-6 membered heterocycloalkyl (substituted by -N(R 4c -1 R 4c ) substituted by -N(R 4c-1 R 4c-2 ) -2 )
- the number of substitutions is 1 or more, for example, 1, 2 or 3
- the heteroatoms in the substituted 3-6 membered heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (eg 1, 2 or 3);
- R 4c-1 and R 4c-2 are independently C 1 -C 4 alkyl
- a a and A b are independently CR 5a or N;
- R 5a is H, C 1 -C 4 alkyl or halogen;
- X a is Y 1 is connected to L a , and Y 2 is connected to L b ;
- n1 is 1, 2 or 3;
- the heteroatom in the 3-9-membered heterocycloalkyl and the 3-9-membered heterocycloalkyl-( CH 2 ) n1 ' is independently N, O or S
- One or more of, the number of heteroatoms is 1-3 (for example, 1, 2 or 3);
- n1' is 1, 2 or 3;
- M 1-1 is C 1 -C 4 alkyl or halogen
- Y 1 is O, -NR 8 or does not exist (that is, M 1 is directly connected to La );
- Ring D is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- n2 is 0, 1, 2 or 3;
- Y 2 is O, -NR 9 or does not exist (that is, the alkylene group in X a is directly connected to L a );
- R 8 and R 9 are independently C 1 -C 4 alkyl
- n3 is 1, 2 or 3;
- n3' is 0, 1, 2 or 3;
- R 10 and R 11 are independently C 1 -C 4 alkyl
- Ring E is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- L a , L b and L c are linking groups
- Q a , Q b and Q c are E3 ubiquitin ligase ligands.
- R 1a , R 1b and R 1c are independently C 6 -C 18 aryl, C 6 -C 18 aryl substituted by R 1a-1 (R 1a-1 is one or more, such as one , 2 or 3) or 5-10 heteroaryl groups, the heteroatoms in the 5-10 heteroaryl groups are independently one or more of N, O or S, and the number of heteroatoms is 1 -4 (e.g. 1, 2, 3 or 4);
- R 1a-1 is independently hydroxy, halogen or C 1 -C 12 alkyl
- R 2a , R 4a , R 2b , R 4b , R 2c and R 5c are independently H or halogen;
- R 3a is a 7-12-membered bridged heterocycloalkyl, the heteroatoms in the bridged heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1 -4 (eg 1, 2, 3 or 4);
- R 4c is H, a 3-6 membered heterocycloalkyl or a 3-6 membered heterocycloalkyl (substituted by -N(R 4c -1 R 4c ) substituted by -N(R 4c-1 R 4c-2 ) -2 )
- the number of substitutions is 1 or more, such as 1, 2 or 3)
- the heteroatom in the 3-6 membered heterocycloalkyl and the heteroatom in the 3- to 6-membered heterocycloalkyl group are replaced by -N(R 4c-1
- the heteroatoms in the 3-6-membered heterocycloalkyl substituted by R 4c-2 ) are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (for example, 1, 2 or 3);
- R 4c-1 and R 4c-2 are independently C 1 -C 4 alkyl
- a a and A b are independently CR 5a or N;
- R 5a is H, C 1 -C 4 alkyl or halogen;
- X a is Y 1 is connected to L a , and Y 2 is connected to L b ;
- n1 is 1, 2 or 3;
- n1' is 1, 2 or 3;
- M 1-1 is C 1 -C 4 alkyl
- Y 1 is O, -NR 8 or does not exist (that is, M 1 is directly connected to La );
- Ring D is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- n2 is 0, 1, 2 or 3;
- Y 2 is O, -NR 9 or does not exist (that is, the alkylene group in X a is directly connected to L a );
- R 8 and R 9 are independently C 1 -C 4 alkyl
- n3 is 1, 2 or 3;
- n3' is 0, 1, 2 or 3;
- R 10 and R 11 are independently C 1 -C 4 alkyl
- Ring E is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- L a , L b and L c are linking groups
- Q a , Q b and Q c are E3 ubiquitin ligase ligands.
- the present invention provides a compound represented by formula Ia, Ib or Ic, its pharmaceutically acceptable salt, its solvate, its stereoisomer, its tautomer , its prodrugs, its metabolites or its isotopic compounds:
- R 1a , R 1b and R 1c are independently C 6 -C 18 aryl, C 6 -C 18 aryl substituted by R 1a-1 (R 1a-1 is one or more, such as 1, 2 or 3) or 5-10 heteroaryl groups, the heteroatoms in the 5-10 heteroaryl groups are independently one or more of N, O or S, and the number of heteroatoms is 1-4 (e.g. 1, 2, 3 or 4);
- R 1a-1 is independently hydroxy, halogen, C 1 -C 12 alkyl or C 2 -C 6 alkynyl;
- R 2a , R 4a , R 2b , R 4b , R 2c and R 5c are independently H or halogen;
- R 3a is a 7-12-membered bridged heterocycloalkyl, the heteroatoms in the bridged heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1 -4 (e.g. 1, 2, 3 or 4);
- R 4c is H, a 3-6 membered heterocycloalkyl or a 3-6 membered heterocycloalkyl (substituted by -N(R 4c -1 R 4c ) substituted by -N(R 4c-1 R 4c-2 ) -2 )
- the number of substitutions is 1 or more, for example, 1, 2 or 3
- the heteroatoms in the substituted 3-6 membered heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (eg 1, 2 or 3);
- R 4c-1 and R 4c-2 are independently C 1 -C 4 alkyl
- a a and A b are independently CR 5a or N;
- R 5a is H, C 1 -C 4 alkyl or halogen;
- X a is Y 1 is connected to L a , and Y 2 is connected to L b ;
- n1 is 1, 2 or 3;
- the heteroatom in the 3-9-membered heterocycloalkyl and the 3-9-membered heterocycloalkyl-( CH 2 ) n1 ' is independently N, O or S
- One or more of, the number of heteroatoms is 1-3 (for example, 1, 2 or 3);
- n1' is 1, 2 or 3;
- M 1-1 is C 1 -C 4 alkyl
- Y 1 is O, -NR 8 or does not exist (that is, M 1 is directly connected to La );
- Ring D is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- n2 is 0, 1, 2 or 3;
- Y 2 is O, -NR 9 or does not exist (that is, the alkylene group in X a is directly connected to L a );
- R 8 and R 9 are independently C 1 -C 4 alkyl
- n3 is 1, 2 or 3;
- n3' is 0, 1, 2 or 3;
- R 10 and R 11 are independently C 1 -C 4 alkyl
- Ring E is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- L a , L b and L c are linking groups
- Q a , Q b and Q c are E3 ubiquitin ligase ligands.
- R 1a , R 1b and R 1c are independently C 6 -C 18 aryl, C 6 -C 18 aryl substituted by R 1a-1 (R 1a-1 is one or more, such as one , 2 or 3) or 5-10 heteroaryl groups, the heteroatoms in the 5-10 heteroaryl groups are independently one or more of N, O or S, and the number of heteroatoms is 1 -4 (e.g. 1, 2, 3 or 4);
- R 1a-1 is independently hydroxy, halogen or C 1 -C 12 alkyl
- R 2a , R 4a , R 2b , R 4b , R 2c and R 5c are independently H or halogen;
- R 3a is a 7-12-membered bridged heterocycloalkyl, the heteroatoms in the bridged heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1 -4 (eg 1, 2, 3 or 4);
- R 4c is H, a 3-6 membered heterocycloalkyl or a 3-6 membered heterocycloalkyl (substituted by -N(R 4c -1 R 4c ) substituted by -N(R 4c-1 R 4c-2 ) -2 )
- the number of substitutions is 1 or more, such as 1, 2 or 3)
- the heteroatom in the 3-6 membered heterocycloalkyl and the heteroatom in the 3- to 6-membered heterocycloalkyl group are replaced by -N(R 4c-1
- the heteroatoms in the 3-6-membered heterocycloalkyl substituted by R 4c-2 ) are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (for example, 1, 2 or 3);
- R 4c-1 and R 4c-2 are independently C 1 -C 4 alkyl
- a a and A b are independently CR 5a or N;
- R 5a is H, C 1 -C 4 alkyl or halogen;
- X a is Y 1 is connected to L a , and Y 2 is connected to L b ;
- n1 is 1, 2 or 3;
- n1' is 1, 2 or 3;
- M 1-1 is C 1 -C 4 alkyl
- Y 1 is O, -NR 8 or does not exist (that is, M 1 is directly connected to La );
- Ring D is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- n2 is 0, 1, 2 or 3;
- Y 2 is O, -NR 9 or does not exist (that is, the alkylene group in X a is directly connected to L a );
- R 8 and R 9 are independently C 1 -C 4 alkyl
- n3 is 1, 2 or 3;
- n3' is 0, 1, 2 or 3;
- R 10 and R 11 are independently C 1 -C 4 alkyl
- Ring E is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- L a , L b and L c are linking groups
- Q a , Q b and Q c are E3 ubiquitin ligase ligands.
- R 1a , R 1b and R 1c are independently C 6 -C 18 aryl, C 6 -C 18 aryl substituted by R 1a-1 (R 1a-1 is one or more, such as 1, 2 or 3) or 5-10 heteroaryl groups, the heteroatoms in the 5-10 heteroaryl groups are independently one or more of N, O or S, and the number of heteroatoms is 1-4 (e.g. 1, 2, 3 or 4);
- R 1a-1 is independently hydroxy, halogen or C 1 -C 12 alkyl
- R 2a , R 4a , R 2b , R 4b , R 2c and R 5c are independently H or halogen;
- R 3a is a 7-12-membered bridged heterocycloalkyl, the heteroatoms in the bridged heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1 -4 (e.g. 1, 2, 3 or 4);
- R 4c is H, a 3-6 membered heterocycloalkyl or a 3-6 membered heterocycloalkyl (substituted by -N(R 4c -1 R 4c ) substituted by -N(R 4c-1 R 4c-2 ) -2 )
- the number of substitutions is 1 or more, such as 1, 2 or 3)
- the heteroatom in the 3-6 membered heterocycloalkyl and the heteroatom in the 3- to 6-membered heterocycloalkyl group are replaced by -N(R 4c-1
- the heteroatoms in the 3-6-membered heterocycloalkyl substituted by R 4c-2 ) are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (for example, 1, 2 or 3);
- R 4c-1 and R 4c-2 are independently C 1 -C 4 alkyl
- a a and A b are independently CR 5a or N;
- R 5a is H, C 1 -C 4 alkyl or halogen;
- X a is Y 1 is connected to L a , and Y 2 is connected to L b ;
- n1 is 1, 2 or 3;
- n1' is 1, 2 or 3;
- M 1-1 is C 1 -C 4 alkyl
- Y 1 is O, -NR 8 or does not exist (that is, M 1 is directly connected to La );
- Ring D is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- n2 is 0, 1, 2 or 3;
- Y 2 is O, -NR 9 or does not exist (that is, the alkylene group in X a is directly connected to L a );
- R 8 and R 9 are independently C 1 -C 4 alkyl
- n3 is 1, 2 or 3;
- n3' is 1, 2 or 3;
- R 10 and R 11 are independently C 1 -C 4 alkyl
- Ring E is a 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number is 1 or 2;
- L a , L b and L c are linking groups
- Q a , Q b and Q c are E3 ubiquitin ligase ligands.
- the C 6 -C 18 aryl group or the C 6 -C 18 aryl group in the C 6 -C 18 aryl group substituted by R 1a-1 C 18 aryl is phenyl, naphthyl, phenanthryl or anthracenyl, preferably naphthyl.
- R 1a , R 1b and R 1c are independently C 6 -C 18 aryl substituted by R 1a -1
- said C 6 -C 18 substituted by R 1a-1 Aryl is wherein R 6 and R 7 are independently H, halogen (eg F, Cl, Br or I), C 1 -C 12 alkyl (eg methyl, ethyl, n-propyl, isopropyl, n-butyl) , isobutyl, sec-butyl or tert-butyl) or C 2 -C 6 alkynyl (eg C 2 -C 4 alkynyl, also eg ethynyl).
- R 6 and R 7 are independently H, halogen (eg F, Cl, Br or I)
- C 1 -C 12 alkyl eg methyl, ethyl, n-propyl, isopropyl, n-butyl
- R 1a , R 1b and R 1c are independently when the said for and / or
- R 1a , R 1b and R 1c are independently when the said for
- R 1a , R 1b and R 1c are independently when the said for
- the halogen is F, Cl, Br or I , such as F or Cl.
- the C 1 -C 12 alkyl group is a C 1 -C 6 alkyl group, and can also be methyl, ethyl, n-propyl, isopropyl propyl, n-butyl, isobutyl, sec-butyl or tert-butyl.
- the C 2 -C 6 alkynyl group is a C 2 -C 4 alkynyl group, and may also be an ethynyl group.
- the 7-12-membered bridged heterocycloalkyl is a 7- or 8-membered bridged heterocycloalkyl
- the heteroatom is N, and the number is 2, or is diazabicyclo[2.2.1]heptyl, diazabicyclo[3.2.1]octyl or diazabicyclo[2.2.2]octyl.
- the 7- to 12-membered bridged heterocycloalkyl is connected to the pyrimidine ring through a heteroatom.
- the 7-12-membered bridged heterocycloalkyl is (E.g ), (E.g )or
- the 3- to 6-membered heterocycloalkyl in the group is azetidinyl, pyrrolidinyl or piperidinyl.
- R 4c the 3-6-membered heterocycloalkyl or the 3-6-membered heterocycloalkane substituted by -N(R 4c-1 R 4c-2 )
- the 3- to 6-membered heterocycloalkyl in the radical is connected to the pyrimidine ring through a heteroatom.
- the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl , sec-butyl or tert-butyl such as methyl.
- the 3-6 membered heterocycloalkyl substituted by -N(R 4c-1 R 4c-2 ) is 1 -N(CH 3 ) 2 )
- Substituted azetidinyl groups such as
- the 3-9 membered heterocycloalkyl is a 3-6 membered monocyclic heterocycloalkyl
- the heteroatoms are independently N and/or O
- the number of heteroatoms is 1 or 2
- further pyrrolidinyl e.g. The a terminal is connected to an alkylene group, and the b terminal is connected to Y 1 ).
- the 3-9-membered heterocycloalkyl in the 3-9-membered heterocycloalkyl substituted by M 1-1 is an 8-9-membered heterocyclic alkyl or 3-6 membered monocyclic heterocycloalkyl, the heteroatoms are independently N and/or O, the number of heteroatoms is 1 or 2, and further is pyrrolidinyl or hexahydro-1H-pyrrolazinyl.
- the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl Butyl such as methyl.
- the halogen is F, Cl, Br or I, such as F.
- the 3-9-membered heterocycloalkyl substituted by M 1-1 is a 3-6-membered monocyclic heterocycloalkyl substituted by methyl, and the heteroatoms are independently is N and/or O, the number of heteroatoms is 1 or 2, and is further a methyl-substituted pyrrolidinyl group (such as The a terminal is connected to an alkylene group, and the b terminal is connected to Y 1 ).
- the 3-9-membered heterocycloalkyl substituted by M 1-1 is hexahydro-1H-pyrrolazinyl substituted by F, and further The a-terminal is connected to an alkylene group, and the b-terminal is connected to Y 1 .
- the 3-6 membered heterocycloalkyl group is an azetidinyl group (such as ), pyrrolidinyl (e.g. ) or piperazine (eg ).
- the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl propyl, n-butyl, isobutyl, sec-butyl or tert-butyl, eg methyl or ethyl.
- the 3-6 membered heterocycloalkyl group is an azetidinyl group (such as ).
- L a , L b and L c are independently The e end is connected with X a , X b or X c , and the f end is connected with Q a , Q b or Q c ;
- n4, n6 and n7 are independently 0, 1 or 2;
- n5 and n8 are independently any integer from 1 to 5 (eg, 1, 2, 3, 4, or 5);
- Y 3 is NH, CH 2 or absent
- n9 is any integer from 0-13 (eg 0, 1, 2 or 12);
- n10 and n13 are independently 0, 1 or 2;
- n11 and n12 are independently any integer from 1 to 7 (eg, 1, 2, 3, 4, or 5);
- Z is CH or N
- Y 5 is a 3-6-membered cycloalkyl, a 3-6-membered heterocycloalkyl or a 5-6-membered heteroaryl, and the heteroatoms in the 3-6-membered heterocycloalkyl are independently One or more of N, O or S, the number of heteroatoms is 1, 2 or 3, and the heteroatoms in the 5-6 membered heterocycloalkyl are independently N, O or S. One or more of , the number of heteroatoms is 1, 2 or 3;
- n14 and n15 are independently 0, 1, 2, 3, 4 or 5;
- n16 and n19 are 0, 1 or 2;
- n17, n18 and n20 are independently 0, 1, 2, 3, 4 or 5;
- Z 1 is CH or N.
- the 3-6 membered cycloalkyl group is preferably cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, such as cyclohexyl.
- the 3-6-membered heterocycloalkyl in Y 5 is preferably a 5-membered heterocycloalkyl whose heteroatom is O or S, and the number is 1, such as furyl
- the 5- to 6-membered heteroaryl group is preferably a 5-membered heteroaryl group whose heteroatom is N and the number is 3, such as 1-H, 1, 2, 3-triazolyl, such as
- L a , L b and L c are independently The e end is connected with X a , X b or X c , and the f end is connected with Q a , Q b or Q c ;
- n4, n6 and n7 are independently 0, 1 or 2;
- n5 and n8 are independently any integer from 1 to 5 (eg, 1, 2, 3, 4, or 5);
- Y 3 is NH, CH 2 or absent
- n9 is any integer from 0-13 (eg 0, 1, 2 or 12);
- n10 and n13 are independently 0, 1 or 2;
- n11 and n12 are independently any integer from 1 to 7 (eg, 2, 3, 4, or 5);
- Z is CH or N
- L a , L b and L c are independently Wherein the left end is connected with X a (X b or N in piperazinyl), and the right end is connected with Q a (Q b or Q c ).
- L a , L b and L c are independently
- Q a , Q b and Q c are independently
- R7 is H or halogen (eg F).
- R 2a , R 2b , R 2c and R 5c are independently halogen (eg, F).
- R 5a is independently halogen (eg, Cl).
- X a is (E.g ), (E.g ), (E.g )or (E.g ).
- n1 is 1 or 3.
- X b is
- n2 is 0 or 1.
- M 1 is absent or 3-9 membered heterocycloalkyl (eg ), Y 1 is -NR 8 .
- M 1 is a 3-9 membered heterocycloalkyl substituted by M 1-1 (for example ), Y 1 is O.
- M 1 is a 3-9 membered heterocycloalkyl (for example ), Y 1 does not exist.
- R 2a is halogen
- R 3a is a 7-12-membered bridged heterocycloalkyl
- the 7-12-membered bridged heterocycloalkyl is a 7- or 8-membered bridged heterocycloalkyl
- the heteroatom is N, and the number is 2 indivual
- R 5a is independently halogen
- R 2a is halogen
- R 3a is a 7-12-membered bridged heterocycloalkyl
- the 7-12-membered bridged heterocycloalkyl is a 7- or 8-membered bridged heterocycloalkyl
- the heteroatom is N, and the number is 2 indivual
- R 5a is halogen
- R 2a is halogen
- R 3a is a 7-12-membered bridged heterocycloalkyl
- the 7-12-membered bridged heterocycloalkyl is a 7- or 8-membered bridged heterocycloalkyl
- the heteroatom is N, and the number is 2 indivual
- R 5a is halogen
- M 1 is a 3-9-membered heterocycloalkyl or a 3-9-membered heterocycloalkyl substituted by M 1-1 ;
- Y 1 is absent
- Ia-1 is Ia-1-1 or Ia-1-2:
- R 5a is independently C 1 -C 4 alkyl or halogen.
- R 5a is halogen
- R 2b is halogen
- X b is n3' is 0;
- n7 0, n8 is 3, 4 or 5, Y3 is CH2 or does not exist;
- n10 and n13 are 0, n11 and n12 are independently 3, 4 or 5, and Y 4 is absent;
- R 5a is halogen
- R 2b is halogen
- the general formula of the compound represented by formula Ib-1 is Ib-1-1 or Ib-1-2:
- R 2a and R 5c are independently halogen
- n4 and n6 are 2, n5 is 4;
- R 4c is a 3-6 membered heterocycloalkyl substituted with -N(R 4c-1 R 4c-2 ).
- R 2a is halogen
- R 5a is halogen
- the compound represented by formula Ia, Ib or Ic is any of the following compounds:
- the compound shown as formula Ia, Ib or Ic is any of the following compounds:
- reaction obtains a compound, the retention time of compound 47-1a is 2.00 minutes under the following chiral resolution conditions: chiral column CHIRALPAK AD-H, 3 x 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide, mobile phase B : isopropanol (0.5%, 2 mol/L ammonia methanol); flow rate: 50 ml/min; column temperature: 35 degrees Celsius; elution with 30% mobile phase B; detector UV 222 nm.
- the salt form of the pharmaceutically acceptable salt of the compound represented by formula Ia, Ib or Ic is formate, trifluoroacetate or hydrochloride.
- the number of salts in the pharmaceutically acceptable salts of the compounds represented by formula Ia, Ib or Ic is preferably 1, 2, 3, 4, 5 or 6.
- the pharmaceutically acceptable salt of the compound represented by formula Ia, Ib or Ic is any of the following compounds:
- the pharmaceutically acceptable salt of the compound represented by formula Ia, Ib or Ic is any of the following compounds:
- the compound represented by formula Ia, Ib or Ic or a pharmaceutically acceptable salt thereof is any of the following compounds:
- the pharmaceutically acceptable salt of the compound represented by formula Ia, Ib or Ic is any of the following compounds:
- the pharmaceutically acceptable salt of the compound represented by formula Ia, Ib or Ic is any of the following compounds:
- hydrochloride salt of the compound obtained by the reaction, the retention time of compound 46-2a is 4.85 minutes under the following chiral conditions: chiral column CHIRAL ART Cellulose-SC, 3 x 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide , mobile phase B: ethanol (0.5%, 2 mol/L ammonia methanol); flow rate: 60 ml/min; column temperature: 35 degrees Celsius; elution with 35% mobile phase B; detector UV222 nm;
- reaction obtains the formate salt of the compound, and the retention time of compound 47-1a is 2.00 minutes under the following chiral resolution conditions: chiral column CHIRALPAK AD-H, 3 x 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide , Mobile phase B: isopropanol (0.5%, 2 mol/liter ammonia methanol); flow rate: 50 ml/min; column temperature: 35 degrees Celsius; elution with 30% mobile phase B; detector UV222 nm.
- the above-mentioned compounds of the present invention represented by formula Ia, Ib or Ic, their pharmaceutically acceptable salts, their solvates, their stereoisomers, their tautomers, their prodrugs, and their metabolites
- the isotopic compounds thereof can be synthesized by methods including methods similar to those known in the chemical field, the steps and conditions of which can be referred to the steps and conditions of similar reactions in the art, especially the synthesis according to the description herein.
- Starting materials are typically from commercial sources such as Aldrich or can be readily prepared using methods well known to those skilled in the art (obtained via SciFinder, Reaxys online database).
- the present invention provides another compound as shown in formula IIa or IIb:
- R 12a and R 12b are independently hydroxyl protecting groups (for example (-MOM)); R 13a and R 13b are independently amino protecting groups (e.g. (-Boc));
- the general formula of the compound represented by the formula IIa or IIb is preferably the following general formula:
- the compound represented by the formula IIa or IIb is preferably any of the following compounds:
- the present invention also provides a pharmaceutical composition
- a pharmaceutical composition comprising substance X and one or more pharmaceutical excipients, wherein the substance X is the above-mentioned compound represented by formula Ia, Ib or Ic, and its pharmaceutically acceptable
- the present invention also provides the use of a substance Y in the preparation of a medicament for the treatment or prevention of cancer mediated by KRAS mutation;
- the substance Y is the above-mentioned compound represented by formula Ia, Ib or Ic, Its pharmaceutically acceptable salt, its solvate, its prodrug, its metabolite or its isotopic compound or the above-mentioned pharmaceutical composition.
- the KRAS mutein is preferably a KRAS_G12D mutein.
- the cancer is preferably blood cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer and the like.
- the present invention also provides the use of a substance Y in the preparation of a medicament for treating or preventing cancer
- the substance Y is the above-mentioned compound represented by formula Ia, Ib or Ic, or a pharmaceutically acceptable salt thereof , its solvate, its stereoisomer, its tautomer, its prodrug, its metabolite or its isotopic compound or the above-mentioned pharmaceutical composition
- the cancer is blood cancer, pancreatic cancer, MYH related Sexual polyposis, colorectal cancer, or lung cancer.
- the present invention also provides a method for treating, preventing or treating cancer mediated by KRAS mutation, comprising administering to a patient a therapeutically effective amount of substance Y; the substance Y is the above-mentioned formula Ia, Ib or Ic
- Said cancer is, for example, blood cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer, and the like.
- the KRAS mutein can be a KRAS_G12D mutein.
- the present invention also provides a method for treating, preventing or treating cancer, which comprises administering to a patient a therapeutically effective amount of substance Y;
- the substance Y is the above-mentioned compound represented by formula Ia, Ib or Ic, its pharmaceutically Acceptable salt, its solvate, its stereoisomer, its tautomer, its prodrug, its metabolite or its isotopic compound or the above-mentioned pharmaceutical composition;
- said cancer is blood cancer, pancreas cancer, MYH-associated polyposis, colorectal cancer, or lung cancer.
- the present disclosure also relates to a method of treating a hyperproliferative disease in a mammal comprising administering to said mammal a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or its derivatives.
- Ras mutations including but not limited to K-Ras, H-Ras or N-Ras mutated Ras mutations that have been identified in hematological cancers or malignancies (eg, cancers affecting the blood, bone marrow, and/or lymph nodes). Accordingly, certain embodiments involve administering a disclosed compound (eg, in the form of a pharmaceutical composition) to a patient in need of treatment of a hematological cancer or malignancy.
- a disclosed compound eg, in the form of a pharmaceutical composition
- the present disclosure relates to methods for treating lung cancer comprising administering to a subject in need thereof an effective amount of any of the foregoing compounds (or pharmaceutical compositions comprising the compounds).
- the cancer or malignant tumor includes but is not limited to leukemia and lymphoma.
- the blood disease is also eg acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic Myeloid leukemia (CML), acute monocytic leukemia (AMoL) and/or other leukemias.
- the lymphoma eg, Hodgkin's lymphoma or all subtypes of non-Hodgkin's lymphoma.
- the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung cancer or large cell lung cancer.
- NSCLC non-small cell lung cancer
- the lung cancer is small cell lung cancer.
- Other lung cancers include, but are not limited to, adenomas, carcinoids, and undifferentiated carcinomas.
- the cancer eg, acute myeloid leukemia, juvenile cancer, childhood adrenal cortical carcinoma, AIDS-related cancers (eg, lymphoma and Kaposi's sarcoma), anal cancer, appendix carcinoma, astrocytoma, atypical malformation, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid, Atypical malformations, embryonal tumors, germ cell tumors, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic Myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors,
- AIDS-related cancers
- pharmaceutically acceptable means that salts, solvents, excipients, etc. are generally non-toxic, safe, and suitable for patient use.
- patient is preferably a mammal, more preferably a human.
- pharmaceutically acceptable salt refers to a pharmaceutically acceptable salt as defined herein, and has all the effects of the parent compound.
- Pharmaceutically acceptable salts can be prepared by treating the organic base in a suitable organic solvent with the corresponding acid according to conventional methods.
- salt formation examples include: for base addition salts, it is possible by using alkali metal or alkaline earth metal hydroxides or alkoxides (such as ethoxide or methoxide) or suitable basic organic amines (such as diethanolamine, bile Alkali (eg, sodium, potassium, or lithium) or alkaline earth (eg, aluminum, magnesium, calcium, zinc, or bismuth) salts are prepared by treating compounds of the invention with appropriate acidic protons with a base or meglumine.
- alkali metal or alkaline earth metal hydroxides or alkoxides such as ethoxide or methoxide
- suitable basic organic amines such as diethanolamine, bile Alkali (eg, sodium, potassium, or lithium)
- alkaline earth salts eg, aluminum, magnesium, calcium, zinc, or bismuth
- salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid; and salts formed with organic acids, such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid Citric acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, glutamic acid, glycolic acid, hydroxynaphthoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, oxalic acid, pyruvic acid, malonate acid, mandelic acid, methanesulfonic acid, mucofuroic acid, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, tartaric acid, citric acid, cinnamic acid, p-toluenesulfonic acid or trimethylace
- organic acids such as ace
- solvate refers to a substance formed by combining a compound of the present invention with a stoichiometric or non-stoichiometric amount of a solvent.
- Solvent molecules in solvates can exist in ordered or non-ordered arrangements.
- the solvent includes, but is not limited to, water, methanol, ethanol, and the like.
- prodrug refers to a compound obtained by modifying the chemical structure of a drug, which is inactive or less active in vitro, and releases the active drug through enzymatic or non-enzymatic transformation in vivo to exert pharmacological effects.
- metabolite refers to intermediate and final metabolites in metabolism.
- isotopic compound refers to a compound in which one or more atoms may exist in their unnatural abundance.
- hydrogen atom its unnaturally abundant form means that about 95% of it is deuterium.
- pharmaceutical excipients may be those excipients widely used in the field of pharmaceutical production. Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide a method to enable the subject to dissolve the active ingredient at a desired rate after administration, or to promote the activity of the subject after the composition is administered. The ingredients are effectively absorbed.
- the pharmaceutical excipients can be inert fillers, or provide some function, such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition.
- Described pharmaceutical adjuvants may include one or more of the following adjuvants: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrating agents, lubricants, anti-sticking agents Agents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavors and sweeteners.
- adjuvants may include one or more of the following adjuvants: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrating agents, lubricants, anti-sticking agents Agents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavors and sweeten
- compositions of the present invention can be prepared in light of the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, attenuating, encapsulating, entrapping or lyophilizing processes.
- compositions of the present invention may be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ophthalmic, rectal, topical or parenteral (infusion, injection, implant Intradermal, subcutaneous, intravenous, intraarterial, intramuscular) administration.
- the pharmaceutical compositions of the present invention may also be in controlled release or delayed release dosage forms (eg, liposomes or microspheres).
- solid oral formulations include, but are not limited to, powders, capsules, caplets, softgels, and tablets.
- liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions.
- topical formulations include, but are not limited to, creams, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum formulations.
- formulations for parenteral administration include, but are not limited to, solutions for injection, dry formulations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection.
- suitable formulations of the pharmaceutical compositions include, but are not limited to, eye drops and other ophthalmic formulations; aerosols: such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories and lozenges agent.
- Treatment means any treatment of a disease in a mammal, including: (1) preventing the disease, i.e. causing the symptoms of the clinical disease not to develop; (2) inhibiting the disease, i.e. preventing the development of the clinical symptoms; (3) alleviating the disease, This results in the subsidence of clinical symptoms.
- an “effective amount” refers to an amount of a compound sufficient to (i) treat the associated disease, (ii) reduce, ameliorate, or eliminate one or more symptoms of a particular disease or disorder, or (iii) when administered to a patient in need of treatment. Delay the onset of one or more symptoms of a particular disease or disorder described herein.
- the amount of the carbonyl heterocyclic compound of formula II, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described above, corresponding to this amount will vary depending on, for example, the particular compound, the disease state and its severity, The characteristics of the patient in need of treatment (eg, weight) and other factors vary, but can nonetheless be routinely determined by those skilled in the art.
- Prevention refers to a reduction in the risk of acquiring or developing a disease or disorder.
- alkyl refers to a straight or branched chain alkyl group having the indicated number of carbon atoms.
- alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and It resembles an alkyl group.
- Alkyl groups are unsubstituted unless a substituent is specifically indicated.
- heterocycloalkyl means a stable 3- to 16-membered saturated cyclic group consisting of 2-11 carbon atoms and 1-5 heteroatoms selected from nitrogen, oxygen and sulfur. Unless specifically stated otherwise in this specification, a heterocycloalkyl group may be monocyclic ("monocyclic heterocycloalkyl"), or a bicyclic, tricyclic or more cyclic ring system, which may include fusion (fused), bridged (bridged), or spiro (spiro) ring systems (e.g., bicyclic ring systems ("bicyclic heterocycloalkyl"). Heterocycloalkyl bicyclic ring systems can be found in One or both rings include one or more heteroatoms; and are saturated. Heterocycloalkyl is unsubstituted unless a substituent is specifically indicated.
- aryl refers to a cyclic, unsaturated, monovalent hydrocarbon group having the specified number of carbon atoms (eg, C6 - C18 ), which is monocyclic or polycyclic (eg, 2 or 3) , when it is a polycyclic ring, two atoms and one bond are shared between the single rings, and (at least one ring/each ring) is aromatic, such as phenyl, naphthyl.
- heteroaryl refers to an aromatic group containing a heteroatom, preferably containing 1, 2 or 3 aromatic 5-6 membered monocyclic or 9-10 membered bicyclic rings independently selected from nitrogen, oxygen and sulfur,
- furanyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl isoxazolyl, oxazolyl, diazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl , thiazolyl, isothiazolyl, thiadiazolyl, benzimidazolyl, indolyl, indazolyl, benzothiazolyl, benziisothiazolyl, benzoxazolyl, benzisoxazolyl, quinoline base, isoquinolyl, etc.
- the present invention describes the structural formula used in refers to the chemical bond that represents the axial chiral stereoconfiguration such as compounds means that
- the present invention describes the group used in the structural formula It means that the corresponding group is connected with other fragments and groups in the compound through this site.
- the reagents and raw materials used in the present invention are all commercially available.
- the positive improvement effect of the present invention is that the compound of the present invention has a degrading effect on KRAS protein and a good inhibitory effect on KRAS G12D mutant protein.
- the compounds and their salts (or free bases) finally prepared in the following examples, if there is a stereo configuration generated by axial chirality in these compounds, these compounds and their salts (or free bases) are formed by axial chirality.
- the resulting stereoconfigurations are consistent with those of the chiral axis-containing intermediates from which these compounds were prepared.
- compounds 1a and 1b and their free bases in Example 1 For example, compounds 1a and 1b and their free bases in Example 1: Compound 1a and its free bases 1a-f (compound 1b and its free bases 1b-f) pass through intermediates 1-9a containing chiral axes (intermediate 1- 9b), the configurations of compound 1a and its free bases 1a-f (compound 1b and its free bases 1b-f) resulting from axial chirality are the same as those of intermediates 1-9a (intermediates 1-9b) same type. The configurations generated by axial chirality in other embodiments of the present invention are all the same as those of embodiment 1.
- N-(4-bromobutyl)phthalimide (10.0 g, 33.7 mmol, 1.0 equiv)
- acetonitrile 250.0 mL
- tert-Butylpiperazine-1-carboxylic acid 7.3 g, 39.0 mmol, 1.1 equiv
- N,N-diisopropylethylamine 9.2 g, 70.9 mmol, 2.0 equiv
- reaction solution was directly purified by reversed-phase flash chromatography (C18 column), and eluted with 5% ⁇ 60% acetonitrile/water mobile phase (0.1% formic acid) within 25 minutes; detector, UV254 nanometer ; yielded compound 1-3 (yellow-green oil, 374 mg, 17% yield).
- the reaction solution was diluted with water (200 mL) and the mixture was extracted with ethyl acetate (3 x 100 mL). The organic layer was washed with saturated brine (1 x 200 mL), followed by drying over anhydrous sodium sulfate. The insolubles were removed by filtration, and the organic phase was concentrated under reduced pressure to obtain the crude product.
- the crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 10% ⁇ 20% ethyl acetate/petroleum ether, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 1-7 (yellow solid, 5.5 g, yield 84%).
- reaction solution was stirred and reacted at 90 degrees Celsius for 2 hours, and the reaction process was monitored by liquid mass and thin layer chromatography. After the reaction was completed, the reaction solution was cooled to 25 degrees Celsius.
- the reaction solution was concentrated and purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 20% ethyl acetate/petroleum ether, and the obtained fraction was evaporated under reduced pressure to remove the solvent to obtain compound 1-9 (two stereoisomers). mixture, white oil, 475.0 mg, 67% yield).
- step 1
- N-(5-Bromopentyl)phthalimide (5.0 g, 16.8 mmol, 1.0 equiv) and 1-tert-butoxycarbonylpiperazine (3.46 g, 1.86 mmol) were combined under stirring at 25 degrees Celsius.
- mol, 1.1 equiv was dissolved in 100 mL of acetonitrile, then N,N-diisopropylethylamine (4.36 g, 33.8 mmol, 2.0 equiv) was added.
- the mixture was warmed to 80 degrees Celsius and stirred at this temperature for 16 hours, the progress of the reaction being monitored by liquid quality. After the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product.
- the obtained crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fraction was evaporated under reduced pressure to remove the solvent to obtain compound 2-1 (brown oil, 6.6 g) , yield 97%).
- reaction solution was purified by reverse-phase flash chromatography column (C18 column), and eluted with 5 ⁇ 95% acetonitrile/water mobile phase (0.1% formic acid) within 30 minutes; the detector UV was 254 nm; compound 2-3 ( Yellow oil, 502.3 mg, 37% yield).
- reaction solution was extracted with saturated sodium bicarbonate solution (500 mL x 2), the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product.
- compound 1-7 (12.0 g, 23.7 mmol, 1.0 equiv.), compound 2-6 (4.8 g, 35.6 mmol, 1.50 equiv.) were added to a 500-mL three-necked flask in turn, fluorinated Cesium (7.2 g, 47.4 mmol, 2.0 equiv); N,N-diisopropylethylamine (6.1 g, 47.4 mmol, 2.0 equiv) and N-methylpyrrolidone (240 mL). The resulting mixture was reacted at 100 degrees Celsius for 16 hours, and the progress of the reaction was monitored by liquid mass and thin chromatography.
- reaction solution was cooled to room temperature, filtered, and the filtrate was directly purified by reversed-phase flash chromatography column (C18 column), using 5% ⁇ 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) within 20 minutes. Elution was performed; detector UV 254 nm; compound 2-7 was obtained (yellow oil, 4.5 g, 29% yield).
- compound 2-7 (4.40 g, 7.3 mmol, 1.0 equiv), 4-(4,4,5,5-tetramethyl-1,3) were added to a 250 ml three-necked flask in turn.
- the resulting mixture was reacted at 60 degrees Celsius for 3 hours, and the progress of the reaction was monitored by liquid mass and thin chromatography. After the reaction, the reaction solution was cooled to room temperature, and the excess solvent was removed under reduced pressure to obtain a crude product.
- the crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 12% methanol/dichloromethane, and the obtained fractions were rotary evaporated under reduced pressure to remove the solvent to obtain compound 2-8 (white solid, 2.4 g, yielded). rate 49%).
- the obtained crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 2-10a (white solid, 190 mg, yield 76%).
- reaction solution was purified by reversed-phase flash chromatography column (C18 column), eluted with 5% ⁇ 95% acetonitrile/water (0.1% ammonia water) mobile phase within 25 minutes, and the detector was UV254 nm to obtain Compound 2-11a (white solid, 80 mg, 63% yield).
- reaction solution was poured into 20 ml of water for dilution, extracted with dichloromethane (15 ml x 3), the organic phases were combined, and the combined organic phase was washed with 50 ml of saturated brine, and the organic phase was washed with Dry over sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain crude product 2-12a (80 mg, yield 88%), which is directly used in the next reaction.
- Preparation conditions reversed-phase column XBridge Shield RP18 OBD Column, 19 x 150 mm, 5 microns; mobile phase A : water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elute with 8%-35% mobile phase B for 7 minutes; detector UV254/220 nm; obtain compound 2 (yellow solid, 30 mg, 33.7% yield).
- the obtained crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 12% methanol (2.5% ammonia methanol)/dichloromethane, and the obtained fraction was evaporated under reduced pressure to remove the solvent to obtain compound 3-1 ( Yellow oil, 3.3 g, 54% yield).
- reaction solution was directly purified by reversed-phase chromatography column (C18 column), eluted with 5% ⁇ 95% methanol/water (0.1% ammonium bicarbonate) mobile phase within 25 minutes, and the detector was UV254/220 nm to obtain Compound 3-3 (red oil, 1.4 g, 52% yield).
- the resulting mixture was stirred and reacted at 60 degrees Celsius for 2 hours, and the reaction progress was monitored by liquid mass and thin layer chromatography. After the reaction was completed, the mixture was cooled to 25°C and concentrated under reduced pressure to obtain the crude product.
- the obtained crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 3-4 (red oil, 1.2 g , the yield is 78%).
- the compound 3-4 (1.2 g) obtained in step 4 was subjected to chiral separation, and the separation conditions were: chiral column NB-Lux 5 ⁇ m i-Cellulose-5, 2.12 x 25cm, 5 ⁇ m); mobile phase A : n-hexane/dichloromethane (5/1) (0.5% 2 mol per liter ammonia methanol), mobile phase B: ethanol; flow rate: 20 ml/min; column temperature: 35 degrees Celsius; use 0% ⁇ 20% mobile phase B eluted for 20 minutes; detector UV 220/254 nm; yielded two products.
- reaction solution was directly purified by reversed-phase chromatography column (C18 column), and eluted with 5% ⁇ 95% methanol/water (0.1% ammonia water) mobile phase within 25 minutes; detector UV254/220 nm; Compound 3 was obtained -5a (brown oil, 92.0 mg, 88% yield).
- compound 2-4 Under stirring at 25 degrees Celsius, compound 2-4 (80.6 mg, 0.15 mmol, 1.0 equiv), sodium cyanoborohydride (29.5 mg, 0.5 mmol, 3.0 equiv) and methanol ( 3.0 ml). Then compound 3-6a (105.1 mg, 0.2 mmol, 1.0 equiv) was dissolved in 2.0 mL of methanol and slowly added dropwise to the reaction solution. The resulting mixture was stirred and reacted at 25 degrees Celsius for 1 hour, and the reaction progress was monitored by liquid mass and thin layer chromatography. After the reaction, the reaction solution was concentrated to obtain a crude product.
- the crude product was purified by high performance liquid phase, preparation conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml /min; eluted with 17% ⁇ 43% mobile phase B for 7 min; detector UV 254/220 nm; product 3-7a was obtained (yellow oil, 110.0 mg, 66% yield).
- the crude product was purified by high performance liquid phase, preparation conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml /min; eluted with 5% ⁇ 16% mobile phase B for 7 min; detector UV 254/220 nm; yielded product 3a (yellow solid, 27 mg, 26% yield).
- reaction solution was directly purified by reversed-phase chromatography column (C18 column), and eluted with 5% ⁇ 95% methanol/water (0.1% ammonia water) mobile phase within 25 minutes; detector UV254/220 nm; Compound 3 was obtained -5b (brown oil, 100.0 mg, 80% yield).
- the crude product was purified by high performance liquid phase, preparation conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml /min; eluted with 5% ⁇ 16% mobile phase B for 7 min; detector UV 254/220 nm; yielded product 3b (yellow solid, 30.0 mg, 23% yield).
- compound 4-1 (1.2 g, 7.5 mL) was sequentially added moles, 2.0 equiv), cesium fluoride (1.2 g, 7.5 mmol, 2.0 equiv) and N-diisopropylethylamine (1.5 g, 11.3 mmol, 3.0 equiv).
- the reaction solution was reacted at 120 degrees Celsius for 2 hours, and the reaction process was monitored by liquid mass and thin layer chromatography. After the reaction was completed, the mixture was cooled to 25 degrees Celsius.
- compound 4-2 (1.0 g, 1.5 mmol, 1.0 equiv.) and 4-(4,4,5,5-tetramethyl-1,3 were added to a 100-ml single-necked flask in sequence.
- the resulting mixture was stirred and reacted at 60 degrees Celsius for two hours, and the reaction progress was monitored by liquid mass and thin layer chromatography. After the reaction was completed, the mixture was cooled to 25°C and concentrated under reduced pressure to obtain the crude product.
- the obtained crude product was purified by silica gel column chromatography, the mobile phase was eluted with a gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 4-3 (red oil, 800.0 mg , yield 77%).
- the compound 4-3 (800 mg) obtained in step 3 was subjected to chiral resolution, and the resolution conditions were: chiral column CHIRALPAK IA, 2 x 25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/ per liter of ammonia methanol), mobile phase B: isopropanol; flow rate: 20 ml/min; elution with 30% mobile phase B for 22 minutes; detector UV 220/210 nm; two products were obtained.
- reaction solution was directly purified by reversed-phase chromatography column (C18 column), and eluted with 5% ⁇ 95% methanol/water (0.1% ammonia water) mobile phase within 25 minutes; detector UV254/220 nm; Compound 4 was obtained -4a (red solid, 180.0 mg, 86% yield).
- the crude product was purified by high performance liquid phase, preparation conditions: high pressure reverse phase column XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 mL/min; eluted with 25%-54% mobile phase B for 7 min; detector UV 254/220 nm; yielded product 4a (yellow solid, 38.0 mg, 46% yield).
- reaction solution was directly purified by reversed-phase chromatography column (C18 column), and eluted with 5% ⁇ 95% methanol/water mobile phase (0.1% ammonia water) within 25 minutes; detector UV254/220 nm; Compound 4 was obtained -4b (red solid, 170.0 mg, 81% yield).
- the crude product was purified by high performance liquid phase, preparation conditions: high pressure reverse phase column XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 mL/min; eluted with 25% ⁇ 54% mobile phase B for 7 min; detector UV 254/220 nm; yielded product 4b (yellow solid, 52.0 mg, 78% yield).
- step 1
- the mixture was reacted under stirring at 25 degrees Celsius for 2 hours, and the reaction process was monitored by liquid quality. After the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product.
- the obtained crude product was purified by silica gel column chromatography (the mobile phase was eluted with a gradient of 0% ⁇ 20% ethyl acetate/dichloromethane), and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 5-1 (yellow solid, 1.16 g, 97% yield).
- reaction solution was cooled to 25 degrees Celsius, and the reaction solution was purified by reverse-phase flash chromatography column (C18 column), and eluted with 10% ⁇ 95% methanol/water (0.1 ammonia water) mobile phase within 20 minutes. ; Detector UV 254 nm; Compound 5-2 was obtained (white solid, 670 mg, 49% yield).
- reaction solution was cooled to 25 degrees Celsius, and concentrated under reduced pressure to obtain a crude product.
- the obtained crude product was purified by silica gel column chromatography (the mobile phase was eluted with a gradient of 0% ⁇ 10% methanol/dichloromethane), and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 5-3 (white solid, 480 mg) , the yield is 68%).
- the obtained crude product was purified by high performance liquid phase, and the preparation conditions were: high pressure reversed-phase column XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; Flow rate: 25 ml/min; elution with 5% ⁇ 20% mobile phase B for 10 min; detector UV 254/220 nm; yield compound 5a (yellow solid, 20 mg, 25% yield).
- reaction solution was purified by reverse-phase flash chromatography column (C18 column), and eluted with a mobile phase of 5% ⁇ 95% methanol/water (0.1% ammonia water) within 20 minutes; the detector UV254 nanometer; obtained Compound 5-4b (white solid, 145 mg, 86% yield).
- the obtained crude product was purified by high performance liquid phase, and the preparation conditions were: high pressure reverse phase column XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.05% trifluoroacetic acid), mobile phase B: Methanol; flow rate: 25 ml/min; eluted with 27% ⁇ 54% mobile phase B for 7 min; detector UV 254/220 nm; yielded compound 5b (yellow solid, 12.6 mg).
- Embodiment 6 (synthesis method 1)
- step 1
- N-(6-bromohexyl)phthalic acid 5.0 g, 16.1 mmol, 1.0 equiv.
- 1-tert-butoxycarbonylpiperazine 3.3 g, 17.7 mmol, 1.1 equiv
- acetonitrile 100 mL
- N,N-diisopropylethylamine 4.17 g, 32.2 mmol, 2.0 equiv.
- the resulting mixture was reacted at 80 degrees Celsius for 16 hours.
- reaction solution was cooled to 25 degrees Celsius.
- the reaction solution was purified by reverse-phase flash chromatography column (C18 column), and eluted with 5% ⁇ 50% methanol/water (0.1% trifluoroacetic acid) mobile phase within 25 minutes; detector UV254 nm; Compound 6 was obtained -3 (yellow solid, 360 mg, 18% yield).
- 6-hepten-1-ol (1.0 g, 8.3 mmol, 1.0 equiv) was dissolved in 30 mL of dichloromethane with stirring at 0°C, then 1,1,1-triacetoxyl was added in portions yl-1,1-dihydro-1,2-phenyliodooxy-3(1H)-one (7.43 g, 17.5 mmol, 2.0 equiv). The resulting mixture was reacted at 25 degrees Celsius for 2 hours, and the progress of the reaction was monitored by thin layer chromatography.
- reaction solution was purified by reverse-phase flash chromatography column (C18 column), and eluted with 5% ⁇ 95% methanol/water (0.1% ammonia water) mobile phase within 25 minutes; detector UV254 nanometer; obtained Compound 6-7a (yellow oil, 77 mg, 73% yield).
- the obtained crude product was purified by high performance liquid phase, and the preparation conditions were: high pressure reversed-phase column XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; Flow rate: 25 mL/min; elution with 5% ⁇ 40% mobile phase B for 7 min; detector UV 254/220 nm; yield compound 6 (yellow solid, 20 mg, 31% yield).
- reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain a crude product.
- the crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 7-2 (bright yellow oil, 2.5 g , the yield is 86%).
- reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain a crude product.
- the crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 7-4 (yellow solid, 200 mg, produced rate 20%).
- reaction solution is heated to 25 degrees Celsius, the reaction solution is concentrated and purified by reversed-phase flash chromatography (C18 column), and 10% ⁇ 95% acetonitrile/water mobile phase (0.1% ammonia water) is used within 20 minutes. Elution was performed; detector UV 254 nm; compound 7-8a was obtained (yellow oil, 80 mg, 81% yield). MS (ESI, m/z): 740.5/742.5 [M+H] + .
- the obtained crude product was purified by high performance liquid phase, and the preparation conditions were: high pressure reversed-phase column XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; Flow rate: 25 ml/min; elution with 5% ⁇ 40% mobile phase B for 10 min; detector UV 254/220 nm; compound 7 (yellow solid, 61.2 mg, 37% yield) was obtained.
- the crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 8-2 (white solid, 80 mg, produced rate 11%).
- the crude product was concentrated under reduced pressure to obtain the crude product.
- the crude product was purified by silica gel column chromatography and eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane. The obtained fractions were evaporated under reduced pressure to remove the solvent to obtain the compound 8-4a (white solid, 150.0 mg, 71% yield).
- reaction solution was directly purified by reversed-phase flash chromatography column (C18 column), and eluted with 30% ⁇ 95% methanol/water mobile phase (0.1% ammonia water) within 20 minutes; detector UV254 nanometer; Compound 8-5a was obtained (pale yellow solid, 66.0 mg, 96% yield).
- the crude product was purified by preparative HPLC, purification conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 mL/min; gradient: 5% ⁇ 19% phase B gradient elution over 10 min; detector UV 254/220 nm; yielded compound 8 (white solid, 15.0 mg, 24% yield).
- the crude product was purified by reverse phase chromatography (C18), eluting with 5% ⁇ 70% acetonitrile/water (0.1% trifluoroacetic acid) mobile phase within 20 minutes, detector UV 254/220 nm, to give the compound 9-5 (yellow solid, 435 mg, 90% yield).
- the crude product was purified by preparative HPLC, purification conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 mL/min; gradient: 5% ⁇ 25% phase B gradient elution over 7 min; detector UV 254/220 nm; yielded compound 9a (yellow solid, 25 mg, 50% yield).
- the crude product was purified by preparative HPLC, purification conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 mL/min; gradient: 5% ⁇ 25% phase B gradient elution over 7 min; detector UV 254/220 nm; yielded compound 9b (yellow solid, 55 mg, 58% yield).
- reaction solution was directly purified by reversed-phase flash chromatography (C18 column), and eluted with 5% ⁇ 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) within 30 minutes; the detector UV254 nm; the compound was obtained 10-1 (white solid, 760 mg, 48% yield).
- the resulting mixture was stirred and reacted at 60 degrees Celsius for 3 hours, and the reaction progress was monitored by liquid mass and thin layer chromatography. After the reaction was completed, the reaction solution was cooled to 25 degrees Celsius, and concentrated under reduced pressure to obtain a crude product.
- the crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 10-2 (yellow solid, 750 mg, produced rate 98%).
- the crude product was purified by preparative HPLC, purification conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate : 25 mL/min; gradient: 35% ⁇ 60% phase B gradient elution over 7 min; detector UV 254/220 nm; yielded 10a (yellow-green solid, 15 mg, 32% yield).
- the crude product was purified by reversed-phase flash chromatography (C18 column), eluting with 5% ⁇ 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) within 30 minutes; detector UV 254 nm; compound 10 was obtained -3b (yellow-green solid, 150 mg, 65% yield).
- reaction solution was cooled to room temperature, and the reaction solution was purified by reverse-phase chromatography (C18), and eluted with 5% ⁇ 95% acetonitrile/water (0.1% trifluoroacetic acid) mobile phase within 25 minutes, Detector: UV 254/220 nm.
- Compound 11-1 was obtained (yellow-green oil, 1.4 g, yield 22%).
- the crude product was purified by preparative HPLC, purification conditions: XSelect CSH Prep C18 OBD Column, 19 x 250 mm, 5 microns; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 mL/min; gradient: 5% ⁇ 24% phase B gradient elution over 7 min; detector UV 254/220 nm; yielded compound 11 (yellow solid, 15.0 mg, 44% yield).
- the resulting mixture was reacted at 25 degrees Celsius for 2 hours, and the progress of the reaction was monitored by liquid mass and thin layer chromatography. After the reaction was completed, the crude product was obtained by concentration under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 35% ethyl acetate/petroleum ether, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 12-1 (yellow solid, 1.9 g, 85% yield).
- reaction solution was directly purified by reversed-phase flash chromatography column (C18 column), and eluted with 50% ⁇ 95% methanol/water mobile phase (0.1% ammonia water) within 20 minutes; the detector UV254 nm; Compound 12- 2 (pale yellow solid, 750.0 mg, 42% yield).
- the resulting mixture was stirred and reacted at 60 degrees Celsius for 1.5 hours, and the reaction progress was monitored by liquid mass and thin layer chromatography. After the reaction was completed, the reaction solution was cooled to 25 degrees Celsius, and concentrated under reduced pressure to obtain a crude product.
- the crude product was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% ⁇ 10% methanol/dichloromethane, and the obtained fractions were evaporated under reduced pressure to remove the solvent to obtain compound 12-3 (pale yellow solid, 673.0 mg, 86% yield).
- the compound 12-3 (673.0 mg) obtained in step 3 was chiral resolved by preparative chiral high-performance liquid chromatography: chiral column CHIRALPAK IA, 2 x 25 cm, 5 ⁇ m; mobile phase A: n-hexane (10 mmol/l ammonia-methanol), mobile phase B: isopropanol; flow rate: 20 ml/min; elution with 50% phase B in 22 min; detector UV 220/254 nm, two products .
- reaction solution was directly purified by reversed-phase flash chromatography column (C18 column), and eluted with 50% ⁇ 95% methanol/water mobile phase (0.1% ammonia water) within 20 minutes; detector UV254 nanometer; Compound 12-4a was obtained (grey solid, 184 mg, 56% yield).
- reaction solution was directly purified by reversed-phase flash chromatography column (C18 column), and eluted with 50% ⁇ 95% methanol/water mobile phase (0.1% ammonia water) within 20 minutes; detector UV254 nanometer; Compound 12-4b was obtained (gray solid, 200.0 mg, 77% yield).
Abstract
一种靶向蛋白调节剂的化合物及其应用。该化合物为如式Ia、Ib或Ic所示的化合物。该化合物对KRAS_G12D突变体具有良好的抑制作用和蛋白降解作用。
Description
本申请要求申请日为2021年4月30日的中国专利申请2021104858187、2021年9月10日的中国专利申请2021110643873、2021年12月24日的中国专利申请2021115999742和2022年1月30日的中国专利申请2022101147173的优先权。本申请引用上述中国专利申请的全文。
本发明涉及一种靶向蛋白调节剂的化合物及其应用。
RAS表示一群具有189个氨基酸的密切相关的单体球状蛋白(21kDa分子量),其与质膜相关并结合GDP或GTP。RAS充当分子开关。当RAS含有结合的GDP时,其处于静止或关闭位置且“无活性”。对细胞暴露于某些生长促进刺激物进行响应时,RAS被诱导以将其结合的GDP交换为GTP。在结合GTP的情况下,RAS被“打开”并能够与其他蛋白(其“下游靶标”)相互作用并活化所述蛋白。RAS蛋白本身具有极低的使GTP水解回GDP、由此使其本身变成关闭状态的固有能力。关闭RAS需要称为GTP酶活化蛋白(GAP)的外源性蛋白,其与RAS相互作用且极大地加速GTP转化成GDP。RAS中影响其与GAP相互作用或使GTP转化回GDP的能力的任何突变将导致蛋白活化延长,且因此传导至细胞告诉其继续生长和分裂的信号延长。由于这些信号导致细胞生长和分裂,因此过度活化的RAS信号传导可最终导致癌症。
在结构上,RAS蛋白包含负责RAS的酶促活性——鸟嘌呤核苷酸结合和水解(GTP酶反应)的G结构域。其还包含称为CAAX盒的C端延伸区,其可被转译后修饰并负责使该蛋白靶向膜。G结构域在尺寸上大约为21-25kDa并含有磷酸结合环(P-环)。P-环表示蛋白中结合核苷酸的囊袋,并且这是具有保守氨基酸残基的结构域的刚性部分,所述保守氨基酸残基为核苷酸结合和水解所必需的(甘氨酸12、苏氨酸26和赖氨酸16)。G结构域还含有所谓的开关I区(残基30-40)和开关II区(残基60-76),其均为蛋白的动态部分,由于该动态部分在静止和负载状态之间进行转换的能力而常常被表示为“弹簧加载”机制。主要相互作用为由苏氨酸-35和甘氨酸-60与GTP的γ-磷酸所形成的氢键,其使开关1区和开关2区分别维持它们的活性构象。在水解GTP和释放磷酸盐之后,此两者松弛成无活性的GDP构象。
RAS亚家族中最值得注意的成员为HRAS、KRAS和NRAS,其主要牵涉许多类型的癌症。然而,存在许多其他成员,包括DIRAS1;DIRAS2;DIRAS3;ERAS;GEM;MRAS;NKIRAS1;NKIRAS2;NRAS;RALA;RALB;RAP1A;RAP1B;RAP2A;RAP2B;RAP2C;RASD1;RASD2;RASL10A;RASL10B;RASL11A;RASL11B;RASL12;REM1;REM2;RERG;RERGL;RRAD;RRAS和RRAS2。
RAS基因的三种主要的同种型(HRAS、NRAS或KRAS)中的任一个的突变为人类肿瘤形成中最常见的事件之一。发现所有的人类肿瘤中约30%的肿瘤在RAS基因中携有一些突变。值得注意的是, 在25%-30%的肿瘤中检测到KRAS突变。相比之下,NRAS和HRAS家族成员中发生致癌突变的比率低得多(分别为8%和3%)。最常见的KRAS突变发现在P-环中的残基G12和G13处以及残基Q61处。在肿瘤相关KRAS G12突变中,KRAS G12D的突变发生概率占比最高,约40%。
基于KRAS异常激活在癌症进展中的重要性和KRAS基因突变在人类癌症中的普遍性,KRAS一直是药物开发人员关注的靶点。尽管已在这个领域中取得进展,但在本领域中仍需要改进的KRAS G12D突变蛋白抑制剂。
近年来,人们利用泛素蛋白酶体通路具有特异性降解蛋白底物的功能特点,构建出靶向泛素化蛋白降解嵌合型分子(proteolysis-targeting chimeras,PROTAC),PROTAC是具有两个异功能的配体通过linker连接的化合物:一个配体靶向于目的蛋白(POI),而另一个配体特异性地募集E3连接酶。当PROTAC结合E3连接酶和目的蛋白时形成了三元复合物,通过劫持E3连接酶,PROTAC使POI呈现出有利的空间位置以促进其泛素化,从而选择性地降低靶蛋白的水平。这种方法的优点是PROTAC可以催化样的多轮降解靶蛋白,这是PROTAC分子与小分子抑制剂最大的不同。
CN110684015A公开了一种靶向ALK的PROTAC分子,成功制备得到了靶向ALK的PROTAC分子,能有效靶向于目标蛋白,并降低细胞中ALK的含量,同时具有较好的体内体外抗肿瘤活性,对正常细胞毒性较低,符合高效低毒的特征。但本领域尚未报道靶向KRAS G12D的PROTAC分子。
发明内容
本发明的所要解决的技术为克服现有技术中能靶向降解突变KRAS的化合物的种类较少的缺陷,为此,提供了一种PROTAC小分子化合物及其应用。本发明的化合物对KRAS_G12D突变体具有良好的抑制作用。
本发明通过以下技术方案解决上述技术问题的。
本发明提供了一种如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:
其中,R
1a、R
1b和R
1c独立地为C
6-C
18芳基、被R
1a-1取代的C
6-C
18芳基(R
1a-1为1个或多个,例如1、2或3个)或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
1a-1独立地为为羟基、卤素、C
1-C
12烷基或C
2-C
6炔基;
R
2a、R
4a、R
2b、R
4b、R
2c和R
5c独立地为H或卤素;
R
3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或 多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
4c为H、3-6元的杂环烷基或被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基(被-N(R
4c-1R
4c-2)取代的个数为1个或多个,例如1、2或3个),所述的3-6元的杂环烷基中的杂原子和被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
R
4c-1和R
4c-2独立地为C
1-C
4烷基;
A
a和A
b独立地为CR
5a或N;R
5a为H、C
1-C
4烷基或卤素;
n1为1、2或3;
M
1为3-9元杂环烷基、被M
1-1取代的3-9元杂环烷基(M
1-1为1个或多个,例如1个、2个或3个;M
1-1仅取代代的3-9元杂环烷基)、-3-9元杂环烷基-(CH
2)
n1’OC(=O)-或不存在(即X
a中的亚烷基与Y
1直接相连);所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH
2)
n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
n1’为1、2或3;
M
1-1为C
1-C
4烷基或卤素;
Y
1为O、-NR
8或不存在(即M
1与L
a直接相连);
环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
n2为0、1、2或3;
Y
2为O、-NR
9或不存在(即X
a中的亚烷基与L
a直接相连);
R
8和R
9独立地为C
1-C
4烷基;
n3为1、2或3;
n3’为0、1、2或3;
R
10和R
11独立地为C
1-C
4烷基;
环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
L
a、L
b和L
c为连接基团;
Q
a、Q
b和Q
c为E3泛素连接酶配体。
在某一优选方案中,一种如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:
其中,R
1a、R
1b和R
1c独立地为C
6-C
18芳基、被R
1a-1取代的C
6-C
18芳基(R
1a-1为1个或多个,例如1个、2个或3个)或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
1a-1独立地为为羟基、卤素或C
1-C
12烷基;
R
2a、R
4a、R
2b、R
4b、R
2c和R
5c独立地为H或卤素;
R
3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1、2、3或4);
R
4c为H、3-6元的杂环烷基或被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基(被-N(R
4c-1R
4c-2)取代的个数为1个或多个,例如1个、2个或3个),所述的3-6元的杂环烷基中的杂原子和被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
R
4c-1和R
4c-2独立地为C
1-C
4烷基;
A
a和A
b独立地为CR
5a或N;R
5a为H、C
1-C
4烷基或卤素;
n1为1、2或3;
M
1为3-9元杂环烷基、被M
1-1取代的3-9元杂环烷基(M
1-1为1个或多个,例如1、2或3个;M
1-1仅取代代的3-9元杂环烷基)、-3-9元杂环烷基-(CH
2)
n1’OC(=O)-或不存在(即X
a中的亚烷基与Y
1直接相连);所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH
2)
n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
n1’为1、2或3;
M
1-1为C
1-C
4烷基;
Y
1为O、-NR
8或不存在(即M
1与L
a直接相连);
环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
n2为0、1、2或3;
Y
2为O、-NR
9或不存在(即X
a中的亚烷基与L
a直接相连);
R
8和R
9独立地为C
1-C
4烷基;
n3为1、2或3;
n3’为0、1、2或3;
R
10和R
11独立地为C
1-C
4烷基;
环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
L
a、L
b和L
c为连接基团;
Q
a、Q
b和Q
c为E3泛素连接酶配体。
在某一优选方案中,本发明提供了一种如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:
其中,R
1a、R
1b和R
1c独立地为C
6-C
18芳基、被R
1a-1取代的C
6-C
18芳基(R
1a-1为1个或多个,例如1、2或3个)或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
1a-1独立地为为羟基、卤素、C
1-C
12烷基或C
2-C
6炔基;
R
2a、R
4a、R
2b、R
4b、R
2c和R
5c独立地为H或卤素;
R
3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
4c为H、3-6元的杂环烷基或被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基(被-N(R
4c-1R
4c-2)取代的个数为1个或多个,例如1、2或3个),所述的3-6元的杂环烷基中的杂原子和被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
R
4c-1和R
4c-2独立地为C
1-C
4烷基;
A
a和A
b独立地为CR
5a或N;R
5a为H、C
1-C
4烷基或卤素;
n1为1、2或3;
M
1为3-9元杂环烷基、被M
1-1取代的3-9元杂环烷基(M
1-1为1个或多个,例如1个、2个或3 个;M
1-1仅取代代的3-9元杂环烷基)、-3-9元杂环烷基-(CH
2)
n1’OC(=O)-或不存在(即X
a中的亚烷基与Y
1直接相连);所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH
2)
n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
n1’为1、2或3;
M
1-1为C
1-C
4烷基;
Y
1为O、-NR
8或不存在(即M
1与L
a直接相连);
环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
n2为0、1、2或3;
Y
2为O、-NR
9或不存在(即X
a中的亚烷基与L
a直接相连);
R
8和R
9独立地为C
1-C
4烷基;
n3为1、2或3;
n3’为0、1、2或3;
R
10和R
11独立地为C
1-C
4烷基;
环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
L
a、L
b和L
c为连接基团;
Q
a、Q
b和Q
c为E3泛素连接酶配体。
在某一优选方案中,一种如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:
其中,R
1a、R
1b和R
1c独立地为C
6-C
18芳基、被R
1a-1取代的C
6-C
18芳基(R
1a-1为1个或多个,例如1个、2个或3个)或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
1a-1独立地为为羟基、卤素或C
1-C
12烷基;
R
2a、R
4a、R
2b、R
4b、R
2c和R
5c独立地为H或卤素;
R
3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1、2、3或4);
R
4c为H、3-6元的杂环烷基或被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基(被-N(R
4c-1R
4c-2)取代的个 数为1个或多个,例如1个、2个或3个),所述的3-6元的杂环烷基中的杂原子和被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
R
4c-1和R
4c-2独立地为C
1-C
4烷基;
A
a和A
b独立地为CR
5a或N;R
5a为H、C
1-C
4烷基或卤素;
n1为1、2或3;
M
1为3-9元杂环烷基、被M
1-1取代的3-9元杂环烷基(M
1-1为1个或多个,例如1、2或3个;M
1-1仅取代代的3-9元杂环烷基)、-3-9元杂环烷基-(CH
2)
n1’OC(=O)-或不存在(即X
a中的亚烷基与Y
1直接相连);所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH
2)
n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
n1’为1、2或3;
M
1-1为C
1-C
4烷基;
Y
1为O、-NR
8或不存在(即M
1与L
a直接相连);
环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
n2为0、1、2或3;
Y
2为O、-NR
9或不存在(即X
a中的亚烷基与L
a直接相连);
R
8和R
9独立地为C
1-C
4烷基;
n3为1、2或3;
n3’为0、1、2或3;
R
10和R
11独立地为C
1-C
4烷基;
环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
L
a、L
b和L
c为连接基团;
Q
a、Q
b和Q
c为E3泛素连接酶配体。
在某一优选方案中,一种如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:
其中,R
1a、R
1b和R
1c独立地为C
6-C
18芳基、被R
1a-1取代的C
6-C
18芳基(R
1a-1为1个或多个,例如1、2或3个)或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
1a-1独立地为羟基、卤素或C
1-C
12烷基;
R
2a、R
4a、R
2b、R
4b、R
2c和R
5c独立地为H或卤素;
R
3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个(例如1个、2个、3个或4个);
R
4c为H、3-6元的杂环烷基或被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基(被-N(R
4c-1R
4c-2)取代的个数为1个或多个,例如1个、2个或3个),所述的3-6元的杂环烷基中的杂原子和被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
R
4c-1和R
4c-2独立地为C
1-C
4烷基;
A
a和A
b独立地为CR
5a或N;R
5a为H、C
1-C
4烷基或卤素;
n1为1、2或3;
M
1为3-9元杂环烷基、被M
1-1取代的3-9元杂环烷基(M
1-1为1个或多个,例如1、2或3个;M
1-1仅取代代的3-9元杂环烷基)、-3-9元杂环烷基-(CH
2)
n1’OC(=O)-或不存在(即X
a中的亚烷基与Y
1直接相连);所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH
2)
n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个(例如1个、2个或3个);
n1’为1、2或3;
M
1-1为C
1-C
4烷基;
Y
1为O、-NR
8或不存在(即M
1与L
a直接相连);
环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
n2为0、1、2或3;
Y
2为O、-NR
9或不存在(即X
a中的亚烷基与L
a直接相连);
R
8和R
9独立地为C
1-C
4烷基;
n3为1、2或3;
n3’为1、2或3;
R
10和R
11独立地为C
1-C
4烷基;
环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;
L
a、L
b和L
c为连接基团;
Q
a、Q
b和Q
c为E3泛素连接酶配体。
在某一优选方案中,R
1a、R
1b和R
1c中,所述的C
6-C
18芳基或所述的被R
1a-1取代的C
6-C
18芳基中的C
6-C
18芳基为苯基、萘基、菲基或蒽基,优选为萘基。
在某一优选方案中,当R
1a、R
1b和R
1c独立地为被R
1a-1取代的C
6-C
18芳基时,所述的被R
1a-1取代的C
6-C
18芳基为
其中,R
6和R
7独立地为H、卤素(例如F、Cl、Br或I)、C
1-C
12烷基(例如甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基)或C
2-C
6炔基(例如C
2-C
4炔基,又例如乙炔基)。
在某一优选方案中,R
6、R
7、R
1a、R
2a、R
4a、R
2b、R
4b、R
2c、R
5a和R
5c中,所述的卤素为F、Cl、Br或I,例如F或Cl。
在某一优选方案中,R
6、R
7和R
1a中,所述的C
1-C
12烷基为C
1-C
6烷基,还可以为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基。
在某一优选方案中,R
6、R
7和R
1a中,所述的C
2-C
6炔基为C
2-C
4炔基,还可以为乙炔基。
在某一优选方案中,R
3a中,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子为N,个数为2个,还可以为二氮杂双环[2.2.1]庚烷基、二氮杂双环[3.2.1]辛烷基或二氮杂双环[2.2.2]辛烷基。
在某一优选方案中,R
3a中,所述的7-12元的桥环杂环烷基通过杂原子与嘧啶环相连接。
在某一优选方案中,R
4c中,所述的3-6元的杂环烷基或所述的被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基中的3-6元的杂环烷基为氮杂环丁烷基、吡咯烷基或哌啶基。
在某一优选方案中,R
4c中,所述的3-6元的杂环烷基或所述的被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基中的3-6元的杂环烷基通过杂原子与嘧啶环相连接。
在某一优选方案中,R
4c-1和R
4c-2中,所述的C
1-C
4烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基,例如甲基。
某一优选方案中,M
1中,所述的被M
1-1取代的3-9元杂环烷基中的3-9元杂环烷基为8-9元并环杂化烷基或3-6元单环杂环烷基,杂原子独立地为N和/或O,杂原子个数为1个或2个,进一步为吡咯烷基或六氢-1H-吡咯嗪基。
某一优选方案中,M
1-1中,所述的C
1-C
4烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基,例如甲基。
某一优选方案中,M
1-1中,所述的卤素为F、Cl、Br或I,例如F。
在某一优选方案中,M
1中,所述的被M
1-1取代的3-9元杂环烷基为被甲基取代的3-6元单环杂环烷基,杂原子独立地为N和/或O,杂原子个数为1或2个,进一步为甲基取代的吡咯烷基(例如
a端与亚烷基相连,b端与Y
1相连)。
在某一优选方案中,M
1中,所述的-3-9元杂环烷基-(CH
2)
n1’OC(=O)-中的3-9元杂环烷基为8-9元并环杂化烷基,杂原子独立地为N和/或O,杂原子个数为1或2个,进一步为
(a端与
中的亚烷基(即-(CH
2)
n1-)相连,a’端与-3-9元杂环烷基-(CH
2)
n1‘OC(=O)-中的亚烷基(即-(CH
2)
n1’-)相连)。
在某一优选方案中,R
5a、R
8、R
9、R
10、R
11和M
1-1中,所述的C
1-C
4烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基,例如甲基或乙基。
n4、n6和n7独立地为0、1或2;
n5和n8独立地为1-5中任一整数(例如1、2、3、4或5);
Y
3为NH、CH
2或不存在;
n9为0-13中任一整数(例如0、1、2或12);
n10和n13独立地为0、1或2;
n11和n12独立地为1-7中任一整数(例如1、2、3、4或5);
Z为CH
2或N;
Y
4为-C(=O)或不存在;
Y
5为3-6元的环烷基、3-6元的杂环烷基或5-6元的杂芳基,所述的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1、2或3个,所述的5-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1、2或3个;
n14和n15独立地为0、1、2、3、4或5;
n16和n19为0、1或2;
n17、n18和n20独立地为0、1、2、3、4或5;
Z
1为CH或N。
Y
5中,所述的3-6元的环烷基优选为环丙基、环丁基、环戊基或环己基,例如环己基。
n4、n6和n7独立地为0、1或2;
n5和n8独立地为1-5中任一整数(例如1、2、3、4或5);
Y
3为NH、CH
2或不存在;
n9为0-13中任一整数(例如0、1、2或12);
n10和n13独立地为0、1或2;
n11和n12独立地为1-7中任一整数(例如2、3、4或5);
Z为CH
2或N;
Y
4为-C(=O)或不存在。
在某一优选方案中,R
7为H或卤素(例如F)。
在某一优选方案中,R
2a、R
2b、R
2c和R
5c独立地为卤素(例如F)。
在某一优选方案中,当A
a和A
b独立地为CR
5a时,R
5a独立地为卤素(例如Cl)。
在某一优选方案中,n1为1或3。
在某一优选方案中,n2为0或1。
在某一优选方案中,
为
M
1为-3-9元杂环烷基-(CH
2)
n1OC(=O)-或不存在,Y
1为-NR
8;M
1为3-9元杂环烷基,Y
1为不存在;或者,M
1为被M
1-1取代的3-9元杂环烷基,Y
1为O。
在某一优选方案中,所述的如式Ia所示的化合物的通式为式Ia-1:
其中,R
2a为卤素;
R
3a为7-12元的桥环杂环烷基,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子为N,个数为2个,
R
5a独立地为卤素;
M
1为-3-9元杂环烷基-(CH
2)
n1OC(=O)-或不存在,Y
1为-NR
8;或者,M
1为3-9元杂环烷基或被M
1-1取代的3-9元杂环烷基,Y
1为O或不存在;
在某一优选方案中,所述的如式Ia所示的化合物的通式为式Ia-1:
其中,R
2a为卤素;
R
3a为7-12元的桥环杂环烷基,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子 为N,个数为2个,
R
5a为卤素;
在某一优选方案中,所述的如式Ia所示的化合物的通式为式Ia-3:
其中,R
2a为卤素;
R
3a为7-12元的桥环杂环烷基,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子为N,个数为2个,
R
5a为卤素;
M
1为3-9元杂环烷基或被M
1-1取代的3-9元杂环烷基;
Y
1为不存在;
在某一优选方案中,所述的如式Ia-1为Ia-1-1或Ia-1-2:
R
5a独立地C
1-C
4烷基或卤素。
在某一优选方案中,所述的如式Ib所示的化合物的通式为式Ib-1:
其中,R
5a为卤素;
R
2b为卤素;
n7为0,n8为3、4或5,Y
3为CH
2或不存在;
n10和n13为0,n11和n12独立地为3、4或5,Y
4为不存在;
在某一优选方案中,所述的如式Ib所示的化合物的通式为式Ib-1:
其中,R
5a为卤素;
R
2b为卤素;
在某一优选方案中,所述的如式Ib-1所示的化合物的通式为Ib-1-1或Ib-1-2:
在某一优选方案中,所述的如式Ic所示的化合物的通式为式Ic'或Ic”:
优选,所述的式Ic'或Ic”中:
R
2a和R
5c独立地为卤素;
n4和n6为2,n5为4;
R
4c为被-N(R
4c-1R
4c-2)取代的3-6元的杂环烷基。
在某一优选方案中,所述的如式Ia所示的化合物的通式为式Ia-2:
其中,R
2a为卤素;R
5a为卤素。
在某一优先方案中,所述的如式Ia、Ib或Ic所示的化合物为如下任一化合物:
在某一优选方案中,所述的如式Ia、Ib或Ic所示的化合物为如下任一化合物:
通过
反应后得到化合物,化合物40-5a在如下手性条件下的保留时间为8.91分钟:手性柱CHIRALPAK IC,2 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/升氨甲醇),流动相B:乙醇;流速:20毫升/分钟;在21分钟内用10%的B相进行洗脱,检测器UV 230/210纳米;
通过
反应得到,化合物46-2a在如下手性条件下的保留时间为4.85分钟:手性柱CHIRAL ART Cellulose-SC,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:乙醇(0.5%,2摩尔/升氨甲醇);流速:60毫升/分钟;柱温:35摄氏度;用35%流动相B洗脱;检测器UV222纳米;
通过
反应得到化合物,化合物47-1a在如下手性拆分条件下的保留时间为2.00分钟:手性柱CHIRALPAK AD-H,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:异丙醇(0.5%,2摩尔/升氨甲醇);流速:50毫升/分钟;柱温:35摄氏度;用30%流动相B洗脱;检测器UV222纳米。
在某一方案中,所述的如式Ia、Ib或Ic所示的化合物药学上可接受的盐的盐型为甲酸盐、三氟醋酸盐或盐酸盐。
所述的如式Ia、Ib或Ic所示的化合物药学上可接受的盐中盐的个数优选为1个、2个、3个、4个、5个或6个。
在某一方案中,所述的如式Ia、Ib或Ic所示的化合物药学上可接受的盐为如下任一化合物:
在某一方案中,所述的如式Ia、Ib或Ic所示的化合物药学上可接受的盐为如下任一化合物:
在某一方案中,所述的如式Ia、Ib或Ic所示的化合物或其药学上可接受的盐为如下任一化合物:
在某一方案中,所述的如式Ia、Ib或Ic所示的化合物药学上可接受的盐为如下任一化合物:
在某一方案中,所述的如式Ia、Ib或Ic所示的化合物药学上可接受的盐为如下任一化合物:
通过
反应后得到化合物的甲酸盐,化合物40-5a在如下手性条件下的保留时间为8.91分钟:手性柱CHIRALPAK IC,2 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/升氨甲醇),流动相B:乙醇;流速:20毫升/分钟;在21分钟内用10%的B相进行洗脱,检测器UV 230/210纳米;
通过
反应得到的化合物的盐酸盐,化合物46-2a在如下手性条件下的保留时间为4.85分钟:手性柱CHIRAL ART Cellulose-SC,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:乙醇(0.5%,2摩尔/升氨甲醇);流速:60毫升/分钟;柱温:35摄氏度;用35%流动相B洗脱;检测器UV222纳米;
通过
反应得到化合物的甲酸盐,化合物47-1a在如下手性拆分条件下的保留时间为2.00分钟:手性柱CHIRALPAK AD-H,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:异丙醇(0.5%,2摩尔/升氨甲醇);流速:50毫升/分钟;柱温:35摄氏度;用30%流动相B洗脱;检测器UV222纳米。
本发明的上述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物可通过包括与化学领域公知方法相似的方法合成,其步骤和条件可参考本领域类似反应的步骤和条件,特别是根据本文说明进行合成。起始原料通常是来自商业来源,例如Aldrich或可使用本领域技术人员公知的方法(通过SciFinder、Reaxys联机数据库得到)容易地制备。
本发明提供了还一种如式IIa或IIb所示的化合物:
R
2a、R
4a、R
6、R
7、R
12a、R
13a、A
a、X
a、L
a、Q
a、R
2b、R
4b、R
12b、R
13b、A
b、X
b、L
b、Q
b的定义均同前所述。
所述的如式IIa或IIb所示的化合物的通式优选如下通式:
所述的如式IIa或IIb所示的化合物优选为如下任一化合物:
例如
本发明还提供了一种药物组合物,其包括物质X和一种或多种药用辅料,所述的物质X为上述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物。
本发明还提供了一种物质Y在制备用于治疗或预防与KRAS突变所介导的癌症的药物中的应用;所述的物质Y为上述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其前药、其代谢产物或其同位素化合物或上述的药物组合物。
所述的应用中,所述的KRAS突变蛋白优选为KRAS_G12D突变蛋白。
所述的应用中,所述的癌症优选为血液癌症、胰腺癌、MYH相关性息肉病、结肠直肠癌或肺癌等。
本发明还提供了一种物质Y在制备用于治疗或预防癌症的药物中的应用;所述的物质Y为上述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构 体、其前药、其代谢产物或其同位素化合物或上述的药物组合物;所述的癌症为血液癌症、胰腺癌、MYH相关性息肉病、结肠直肠癌或肺癌。
本发明还提供了一种治疗预防或治疗与KRAS突变所介导的癌症的方法,其包括向患者施用治疗有效量的物质Y;所述的物质Y为上述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其前药、其代谢产物或其同位素化合物或上述的药物组合物。
所述的癌症例如血液癌症、胰腺癌、MYH相关性息肉病、结肠直肠癌或肺癌等。
所述的KRAS突变蛋白可为KRAS_G12D突变蛋白。
本发明还提供了一种治疗预防或治疗癌症的方法,其包括向患者施用治疗有效量的物质Y;所述的物质Y为上述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物或上述的药物组合物;所述的癌症为血液癌症、胰腺癌、MYH相关性息肉病、结肠直肠癌或肺癌。
本公开还涉及治疗哺乳动物过度增殖性疾病的方法,该方法包括向所述哺乳动物施用治疗有效量的本公开的化合物或药学上可接受的盐,酯,前药,溶剂化物,水合物或其衍生物。
Ras突变,包括但不限于已经在血液系统癌症或恶性肿瘤(例如影响血液,骨髓和/或淋巴结的癌症)中鉴定出的K-Ras,H-Ras或N-Ras突变的Ras突变。因此,某些实施方案涉及向需要治疗血液癌症或恶性肿瘤的患者施用所公开的化合物(例如,以药物组合物的形式)。
在某些特定实施方案中,本公开涉及用于治疗肺癌的方法,所述方法包括将有效量的任何上述化合物(或包含所述化合物的药物组合物)施用于需要其的受试者。
本发明中,所述的癌症或恶性肿瘤包括但不限于白血病和淋巴瘤。在某些实施方案中,所述的血病又例如急性淋巴细胞白血病(ALL),急性骨髓性白血病(AML),慢性淋巴细胞性白血病(CLL),小淋巴细胞性淋巴瘤(SLL),慢性骨髓性白血病(CML),急性单核细胞白血病(AMoL)和/或其他白血病。在某些实施方案中,所述的淋巴瘤,例如霍奇金淋巴瘤或非霍奇金淋巴瘤的所有亚型。
本发明中,在某些实施方案中,所述的肺癌是非小细胞肺癌(NSCLC),例如腺癌,鳞状细胞肺癌或大细胞肺癌。在其他实施方案中,肺癌是小细胞肺癌。其他肺癌包括但不限于腺瘤,类癌和未分化癌。
本发明中,在一些实施方案中,所述癌症,例如急性髓细胞性白血病,青少年癌症,儿童肾上腺皮质癌,与艾滋病相关的癌症(例如淋巴瘤和卡波济氏肉瘤),肛门癌,阑尾癌,星形细胞瘤,非典型类畸形,基底细胞癌,胆管癌,膀胱癌,骨癌,脑干神经胶质瘤,脑肿瘤,乳腺癌,支气管肿瘤,伯基特淋巴瘤,类癌,非典型类畸形,胚胎肿瘤,生殖细胞肿瘤,原发性淋巴瘤,宫颈癌,儿童期癌症,脊索瘤,心脏肿瘤,慢性淋巴细胞性白血病(CLL),慢性粒细胞性白血病(CML),慢性骨髓增生性疾病,结肠癌,大肠癌,颅咽管瘤,皮肤T细胞淋巴瘤,肝外导管原位癌(DCIS),胚胎肿瘤,中枢神经系统癌,子宫内膜癌,室管膜瘤,食管癌,肉芽肿性神经母细胞瘤,尤文氏肉瘤,颅外生殖细胞瘤,性腺外生殖细胞瘤,眼癌,骨纤维组织细胞瘤,胆囊癌,胃癌,胃肠道类癌,胃肠道间质瘤(GIST),生殖细胞肿瘤,妊娠滋养细胞肿瘤,毛细胞白血病,头颈癌,心脏病,肝癌,霍奇金淋巴瘤,下咽癌, 眼内黑色素瘤,胰岛细胞肿瘤,胰腺神经内分泌肿瘤,肾癌,喉癌,嘴唇和口腔癌,肝癌,小叶原位癌(LCIS),肺癌,淋巴瘤,转移性鳞状鳞癌,隐匿性原发性,中线癌,口腔癌,多发性内分泌肿瘤综合征,多发性骨髓瘤/浆细胞瘤,真菌病,蕈样肉芽肿,骨髓增生异常综合征,骨髓增生异常/骨髓增生性肿瘤,多发性骨髓瘤,默克尔细胞癌,恶性间皮瘤,骨和骨肉瘤的恶性纤维组织细胞瘤,鼻腔和副鼻窦,鼻腔和鼻窦神经母细胞瘤,非霍奇金淋巴瘤,非小细胞肺癌(NSCLC),口腔癌,嘴唇和口腔癌,口咽癌,卵巢癌,胰腺癌,乳头状瘤病,副神经节瘤,鼻旁窦和鼻腔癌,甲状旁腺癌,阴茎癌,咽喉癌,胸膜肺母细胞瘤,原发性中枢神经系统(CNS)淋巴瘤,前列腺癌,直肠癌症,移行细胞癌,视网膜母细胞瘤,横纹肌肉瘤,唾液腺癌,皮肤癌,胃(胃)癌,小细胞肺癌,小肠癌,软组织肉瘤,T细胞淋巴瘤,睾丸癌,喉癌,胸腺瘤和胸腺癌癌,甲状腺癌,肾盂和输尿管的移行细胞癌,滋养细胞肿瘤,儿童期不常见的癌,尿道癌,子宫肉瘤,阴道癌,外阴癌或病毒性癌。在一些实施方案中,所述非癌性过度增殖性疾病,例如皮肤的良性增生(例如牛皮癣),再狭窄或前列腺(例如良性前列腺肥大(BPH))。
术语定义
术语“药学上可接受的”是指盐、溶剂、辅料等一般无毒、安全,并且适合于患者使用。所述的“患者”优选哺乳动物,更优选为人类。
术语“药学上可接受的盐”指的是本文所定义的药学上可接受的盐,并且具有母体化合物所有的作用。药学上可接受的盐可以通过在有机碱的合适的有机溶剂中加入相应的酸,根据常规方法处理来制备药学上可接受的盐。
成盐实例包括:对于碱加成盐,有可能通过在水性介质中使用碱金属或碱土金属氢氧化物或醇盐(例如乙醇盐或甲醇盐)或适当碱性有机胺(例如二乙醇胺、胆碱或葡甲胺)处理具有适当酸性质子的本发明化合物来制备碱金属(如钠、钾或锂)或碱土金属(如铝、镁、钙、锌或铋)的盐。
或者,对于酸加成盐,与无机酸成盐,如盐酸、氢溴酸、硫酸、硝酸、磷酸;和有机酸所形成的盐,如醋酸、苯磺酸、苯甲酸、樟脑磺酸、柃檬酸、乙磺酸、富马酸、葡庚糖酸、谷氨酸、乙醇酸、羟基萘甲酸、2-羟基乙磺酸、乳酸、马来酸、苹果酸、草酸、丙酮酸、丙二酸、扁桃酸、甲磺酸、黏糠酸、2-萘磺酸、丙酸、水杨酸、琥铂酸、酒石酸、柠檬酸、肉桂酸、对甲苯磺酸或三甲基乙酸。
术语“溶剂合物”是指本发明化合物与化学计量或非化学计量的溶剂结合形成的物质。溶剂合物中的溶剂分子可以有序或非有序排列的形式存在。所述的溶剂包括但不限于:水、甲醇、乙醇等。
术语“前药”是指药物经过化学结构修饰后得到的在体外无活性或活性较小、在体内经酶或非酶的转化释放出活性药物而发挥药效的化合物。
术语“代谢产物”是指新陈代谢中的中间代谢产物和最终代谢产物。
术语“同位素化合物”是指化合物中的一个或多个原子可以其非天然丰度的形式存在。以氢原子为例,其非天然丰度的形式是指其中约95%为氘。
术语“药用辅料”可为药物生产领域中广泛采用的那些辅料。辅料主要用于提供一个安全、稳定和功能性的药物组合物,还可以提供方法,使受试者接受给药后活性成分以所期望速率溶出,或促进受 试者接受组合物给药后活性成分得到有效吸收。所述的药用辅料可以是惰性填充剂,或者提供某种功能,例如稳定该组合物的整体pH值或防止组合物活性成分的降解。所述的药用辅料可以包括下列辅料中的一种或多种:粘合剂、助悬剂、乳化剂、稀释剂、填充剂、成粒剂、胶粘剂、崩解剂、润滑剂、抗粘着剂、助流剂、润湿剂、胶凝剂、吸收延迟剂、溶解抑制剂、增强剂、吸附剂、缓冲剂、螯合剂、防腐剂、着色剂、矫味剂和甜味剂。
本发明的药物组合物可根据公开的内容使用本领域技术人员已知的任何方法来制备。例如,常规混合、溶解、造粒、乳化、磨细、包封、包埋或冻干工艺。
本发明所述的药物组合物可以任何形式给药,包括注射(静脉内)、粘膜、口服(固体和液体制剂)、吸入、眼部、直肠、局部或胃肠外(输注、注射、植入、皮下、静脉内、动脉内、肌内)给药。本发明的药物组合物还可以是控释或延迟释放剂型(例如脂质体或微球)。固体口服制剂的实例包括但不限于粉末、胶囊、囊片、软胶囊剂和片剂。口服或粘膜给药的液体制剂实例包括但不限于悬浮液、乳液、酏剂和溶液。局部用制剂的实例包括但不限于乳剂、凝胶剂、软膏剂、乳膏剂、贴剂、糊剂、泡沫剂、洗剂、滴剂或血清制剂。胃肠外给药的制剂实例包括但不限于注射用溶液、可以溶解或悬浮在药学上可接受载体中的干制剂、注射用悬浮液和注射用乳剂。所述的药物组合物的其它合适制剂的实例包括但不限于滴眼液和其他眼科制剂;气雾剂:如鼻腔喷雾剂或吸入剂;适于胃肠外给药的液体剂型;栓剂以及锭剂。
“治疗”意味着对哺乳动物体内疾病的任何治疗,包括:(1)防止疾病,即造成临床疾病的症状不发展;(2)抑制疾病,即阻止临床症状的发展;(3)减轻疾病,即造成临床症状的消退。
“有效量”是指当向需要治疗的患者给予化合物时,其量足以(i)治疗相关疾病,(ii)减弱、改善或消除特定疾病或病症的一种或多种症状,或(iii)延迟本文所述的特定疾病或病症的一种或多种症状的发作。对应于该量的所述的如式II所示的羰基杂环类化合物或其药学上可接受的盐或者如上所述的药物组合物的量将根据例如特定化合物、疾病状况及其严重性、需要治疗的患者的特征(例如体重)等因素而变化,但是尽管如此仍然可以由本领域技术人员常规地确定。
本发明所述的“预防”是指获得或发生疾病或障碍的风险降低。
术语“烷基”是指具有指定的碳原子数的直链或支链烷基。烷基的实例包括甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基、正戊基、正己基、正庚基、正辛基及其类似烷基。若非特别指明取代基,否则烷基是非取代的。
术语“杂环烷基”意指由2-11个碳原子以及1-5个选自氮、氧和硫的杂原子组成的稳定的3元至16元饱和环状基团。除非本说明书中另外特别指明,否则杂环烷基基团可以是单环的(“单环的杂环烷基”),或者是双环、三环或更多环的环体系,其可包括融合的(稠环)、桥联的(桥环)或螺的(螺环)环系统(例如二环系统(“二环的杂环烷基”)。杂环烷基二环的环系统可以在一个或两个环中包括一个或多个杂原子;并且是饱和的。若非特别指明取代基,否则杂环烷基是非取代的。
术语“芳基”是指具有指定碳原子数(例如,C
6~C
18)的、环状的、不饱和的一价烃基,其为单环或多环(例如,2个或3个),为多环时,单环之间共用两个原子和一根键,且(至少一个环/每个 环均)具有芳香性,例如苯基、萘基。
术语“杂芳基”是指含有杂原子的芳香基团,优选含有1个、2个或3个独立选自氮、氧和硫的芳族5-6元单环或9-10元双环,例如呋喃基、吡啶基、哒嗪基、嘧啶基、吡嗪基、噻吩基、异唑基、噁唑基、二唑基、咪唑基、吡咯基、吡唑基、三唑基、四唑基、噻唑基、异噻唑基、噻二唑基、苯并咪唑基、吲哚基、吲唑基、苯并噻唑基、苯并异噻唑基、苯并唑基、苯并异唑基、喹啉基、异喹啉基等。
在不违背本领域常识的基础上,上述各优选条件,可任意组合,即得本发明各较佳实例。
本发明所用试剂和原料均市售可得。
本发明的积极进步效果在于:本发明的化合物对KRAS蛋白具有降解作用和对KRAS G12D突变蛋白具有良好的抑制作用。
下面通过实施例的方式进一步说明本发明,但并不因此将本发明限制在所述的实施例范围之中。下列实施例中未注明具体条件的实验方法,按照常规方法和条件,或按照商品说明书选择。
本发明中,以下实施例最终制得的化合物及其盐(或游离碱),若这些化合物中存在由轴手性产生的立体构型时,这些化合物及其盐(或游离碱)由轴手性产生的立体构型与制备这些化合物的含有手性轴中间体的构型一致。例如实施例1中化合物1a和1b及其游离碱:化合物1a及其游离碱1a-f(化合物1b及其游离碱1b-f)通过含有手性轴的中间体1-9a(中间体1-9b)制得,则化合物1a及其游离碱1a-f(化合物1b及其游离碱1b-f)中由轴手性产生的构型与中间体1-9a(中间体1-9b)的构型一致。本发明的其他实施例中由轴手性产生的构型均同实施例1情况。
游离碱:
另外,本领域技术人员根据上述制备参数,相应的选择和调整柱层析的条件,即可得到中间体1-9a和1-9b的分析参数。
实施例1
4-((4-(4-(3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮甲酸盐1a;4-((4-(4-(3-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮甲酸盐1b。
步骤1
在25摄氏度搅拌条件下,向500毫升圆底烧瓶中依次加入N-(4-溴丁基)邻苯二甲酰亚胺(10.0克,33.7毫摩尔,1.0当量),乙腈(250.0毫升)和叔丁基哌嗪-1-羧酸(7.3克,39.0毫摩尔,1.1当量),然后在25摄氏度下滴加N,N-二异丙基乙胺(9.2克,70.9毫摩尔,2.0当量)。将混合物在80摄氏度下搅拌16小时。反应过程通过液质和薄层层析来监控。反应完成后,降温至25摄氏度,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物1-1(红色油,13.5克,产率98%)。MS(ESI,m/z):388.2[M+H]
+。
步骤2
在25摄氏度,氮气保护搅拌条件下,向500毫升反应瓶中依次加入化合物1-1(5.5克,13.5毫摩尔,1.0当量),乙醇(260.0毫升)和水合肼(3.9克,74.2毫摩尔,5.5当量)。所得混合物在80摄氏度下搅拌反应16小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→20%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物1-2(红色油,3.2克,产率88%)。MS(ESI,m/z):258.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ3.48–3.40(m,4H),2.72(t,J=6.6Hz,2H),2.42–2.32(m,6H),1.60–1.48(m,4H),1.46(s,9H)。
步骤3
在25摄氏度,氮气保护搅拌条件下,向化合物1-2(1.0克,3.5毫摩尔,1.0当量),超干的N-甲 基吡咯烷酮(10.0毫升)和4-(4-氨基丁基)哌嗪-1-羧酸叔丁酯(1.1克,4.1毫摩尔,1.2当量)的混合物中滴加N,N-二异丙基乙胺(0.9克,7.2毫摩尔,2.0当量)。将所得混合物在90摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→60%的乙腈/水流动相(0.1%甲酸)进行洗脱;检测器,UV254纳米;得到化合物1-3(黄绿色油,374毫克,产率17%)。MS(ESI,m/z):514.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.36(s,1H),8.24(s,1H),7.54–7.47(m,1H),7.15–7.09(m,1H),6.89(d,J=8.5Hz,1H),6.25(s,1H),4.99–4.87(m,1H),3.66–3.55(m,4H),3.36–3.28(m,2H),2.95–2.63(m,10H),1.83–1.64(m,4H),1.46(s,9H)。
步骤4
在室温搅拌条件下,向化合物1-3(374.0毫克,0.7毫摩尔,1.0当量)的二氯甲烷(10.0毫升)溶液中加入三氟乙酸(5.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后。减压浓缩除去多余的试剂得到粗产品1-4(黄色固体,300.0毫克,产率88%)。MS(ESI,m/z):414.1[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向500毫升圆底烧瓶中加入2-氨基-4-溴-5-氯-3-氟苯甲酸(15.0克,55.9毫摩尔,1.0当量)和尿素(33.6克,558.7毫摩尔,10.0当量)。将混合物升温至150摄氏度并在该温度下搅拌6小时。反应完成后,降温至25摄氏度,加入750毫升水稀释,搅拌30分钟后过滤,滤饼用水(50毫升 x 3)洗涤,收集固体减压干燥后得到化合物1-5粗产品(黄色固体,15.5克,纯度56%),该化合物不需进一步纯化,直接用于下一步合成。MS(ESI,m/z):290.9/292.9/294.8[M-H]
-;
1H NMR(400MHz,DMSO-d
6)δ7.79(d,J=1.8Hz,1H),6.86(s,1H),5.41(s,1H)。
步骤6
在25摄氏度,氮气保护搅拌条件下,向反应瓶中依次加入化合物1-5(10.0克,纯度56%,19.0毫摩尔,1.0当量),N,N-二异丙基乙胺(15.0毫升,81.8毫摩尔,4.3当量)和三氯氧磷(150.0毫升)。所得混合物在90摄氏度下搅拌反应5小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩除去多余的试剂。然后向所得粗产物中加入400毫升水,用乙酸乙酯(400毫升 x 3)萃取。合并有机相,有机相用300毫升饱和食盐水洗涤,洗涤后有机相用无水硫酸钠干燥,过滤除去干燥剂;滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→12%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物1-6(黄色固体,4.4克,产率70%)。MS(ESI,m/z):328.8/330.8/332.8[M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.21(d,J=2.0Hz,1H)。
步骤7
在零摄氏度,氮气保护搅拌条件下,向化合物1-6(4.14克,12.5毫摩尔,1.0当量),超干的1,4-二氧六环(80.0毫升)和三乙胺(4.00克,37.5毫摩尔,3当量)的混合物中滴加哌嗪-1-甲酸叔丁酯(2.45毫克,12.5毫摩尔,1.0当量)的超干的1,4-二氧六环(15.0毫升)溶液。混合物在25摄氏度下反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,冷却至室温。反应液加水(200毫升)稀释,混合物用乙酸乙酯(3 x 100毫升)萃取。有机层用饱和食盐水(1 x 200毫升)清洗,随后用无水硫酸钠干燥。过滤除去不溶物,有机相减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用10%→20%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物1-7(黄色固体,5.5克,产率84%)。MS(ESI,m/z):479.0/481.0/483.0[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.76(d,J=2.0Hz,1H),3.95–3.83(m,4H),3.71–3.64(m,4H),1.50(s,9H)。
步骤8
在25摄氏度,氮气保护搅拌条件下,向化合物1-7(3.0克,6.0毫摩尔,1.0当量)的N-甲基吡咯烷酮(30.0毫升)溶液中加入3-丁烯-1-醇(1.8克,23.7毫摩尔,4.0当量),N,N-二异丙基乙胺(1.6克,11.9毫摩尔,2.0当量)和氟化铯(1.9克,11.9毫摩尔,2.0当量)。反应液在130摄氏度条件下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度。 用二氯甲烷(300毫升 x 3)萃取所得混合物。将合并的有机层用300毫升水洗涤,用无水硫酸钠干燥。过滤后,将滤液在减压下浓缩。残留物通过硅胶柱色谱纯化,用0→20%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物1-8(白色固体,900毫克,产率29%)。MS(ESI,m/z):515.1/517.1/519.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.70(d,J=2.0Hz,1H),5.97–5.87(m,1H),5.21–5.09(m,2H),4.50(t,J=7.0Hz,2H),3.80–3.73(m,4H),3.68–3.61(m,4H),2.64–2.59(m,2H),1.49(s,9H)。
步骤9
在25摄氏度,氮气保护搅拌条件下,向化合物1-8(620.0毫克,1.2毫摩尔,1.0当量)的1,4-二氧六环/水(5/1,12.0毫升)溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(389.7毫克,1.4毫摩尔,1.2当量),碳酸钠(382.2毫克,3.6毫摩尔,3.0当量)和1,1'-二(二苯膦基)二茂铁二氯化钯(II)(97.9毫克,0.1毫摩尔,0.1当量)。反应液在90摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度。反应液浓缩后通过硅胶柱色谱纯化,用0%→20%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物1-9(二个立体异构体混合物,白色油,475.0毫克,产率67%)。MS(ESI,m/z):579.2/581.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.81–7.76(m,2H),7.51–7.43(m,1H),7.37–7.31(m,2H),7.28–7.23(m,1H),7.12(d,J=2.5Hz,1H),6.02–5.85(m,1H),5.23–5.07(m,2H),4.53(t,J=7.0Hz,2H),3.92–3.82(m,4H),3.77–3.67(m,4H),2.68–2.58(m,2H),1.53(s,9H)。
步骤10
通过制备级手性高效液相色谱法对步骤9所得化合物1-9(475.0毫克)进行手性拆分:手性柱CHIRALPAK IC,2 x 25厘米,5微米;流动相A:正己烷/二氯甲烷=5/1(0.1%二乙胺),流动相B:乙醇;流速:15毫升/分钟;在12分钟内用50%的B相进行洗脱,检测器UV 220/235纳米,得到两个产品。较短保留时间(4.935分钟)的产品为化合物1-9a(白色固体,205.0毫克),化合物1-9a:MS(ESI,m/z):579.2/581.1[M+H]
+;较长保留时间(8.413分钟)的产品为化合物1-9b(白色固体,210.0毫克),化合物1-9b:MS(ESI,m/z):579.2/581.1[M+H]
+。
步骤11
在25摄氏度,搅拌条件下,向化合物1-9a(180.0毫克,0.3毫摩尔,1.0当量)的丙酮/水(4/1,10.0.毫升)溶液中加入N-甲基吗啉氧化物(72.8毫克,0.6毫摩尔,2.0当量),然后在0摄氏度下加入四氧化锇(39.5毫克,0.2毫摩尔,0.5当量)。反应液在25摄氏度条件下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩,所得混合物通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→60%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器,UV254纳米;得到化合物1-10a(白色油,150.0毫克,产率77%)。MS(ESI,m/z):613.3/615.2[M+H]
+。
步骤12
在25摄氏度,氮气保护搅拌条件下,向化合物1-10a(130.0毫克,0.2毫摩尔,1.0当量)的乙腈/水(4/1,10.0毫升)溶液中分批加入高碘酸钠(272.1毫克,1.2毫摩尔,6.0当量)。反应液在25摄氏度条件下搅拌反应0.5小时,反应过程通过液质和薄层层析来监控。反应完全后,用二氯甲烷(10毫升 x 3)萃取。将合并的有机层用10毫升饱和氯化钠溶液洗涤,洗涤后有机相用无水硫酸钠干燥,过滤,滤液减压浓缩,得到化合物1-11a(油,123.0毫克,产率100%)。MS(ESI,m/z):581.2/583.2[M+H]
+。
步骤13
在25摄氏度,搅拌条件下,向化合物1-11a(123.0毫克,0.2毫摩尔,1.0当量)的甲醇(5.0毫 升)溶液中加入化合物1-4(105.0毫克,0.3毫摩尔,1.2当量),然后在25摄氏度下加入冰乙酸(19.1毫克,0.3毫摩尔,1.5当量)和氰基硼氢化钠(20.0毫克,0.3毫摩尔,1.5当量),反应液在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩,所得混合物通过液相色谱进行纯化,色谱柱XSelect CSH Prep C18 OBD,19 x 250毫米,5微米;流动相A:水(0.05%甲酸),流动相B:乙腈;流速:25毫升/分钟;在7分钟内用20%→50%的B相进行洗脱,检测器UV 220/235纳米。得到化合物1-12a(黄绿色固体,40.0毫克,产率19%)。MS(ESI,m/z):978.4/980.4[M+H]
+。
步骤14
在室温搅拌条件下,向化合物1-12a(40.0毫克,0.04毫摩尔,1当量)的二氯甲烷(5.0毫升)溶液中加入三氟乙酸(2.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩,所得混合物通过制备级高效液相色谱进行纯化,色谱柱XSelect CSH Prep C18 OBD,19 x 250毫米,5微米;流动相A:水(0.05%甲酸),流动相B:乙腈;流速:25毫升/分钟;在7分钟内用10%→35%的B相进行洗脱,检测器UV 220/235纳米,得到化合物1a(黄绿色固体,15.0毫克,产率40%)。MS(ESI,m/z):878.4/880.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ11.10(s,1H),8.23–8.18(m,1H),7.97–7.93(m,1H),7.81(d,J=8.3Hz,1H),7.60–7.54(m,1H),7.47–7.41(m,1H),7.29(d,J=2.4Hz,1H),7.26–7.17(m,2H),7.12(d,J=8.6Hz,1H),7.08–7.05(m,1H),7.01(d,J=7.0Hz,1H),6.56(t,J=6.0Hz,1H),5.08–5.01(m,1H),4.36(t,J=6.5Hz,2H),3.86–3.77(m,4H),3.36–3.24(m,4H),3.01–2.94(m,4H),2.93–2.81(m,2H),2.64–2.51(m,2H),2.46–2.23(m,10H),2.07–1.97(m,1H),1.94–1.84(m,2H),1.62–1.42(m,4H)。
步骤11’
在25摄氏度,搅拌条件下,向化合物1-9b(210.0毫克,0.4毫摩尔,1.0当量)的丙酮/水(4/1,10毫升)溶液中加入N-甲基吗啉氧化物(89.4毫克,0.7毫摩尔,2.0当量),然后在0摄氏度下加入四氧化锇(46.1毫克,0.2毫摩尔,0.5当量)。反应液在25摄氏度条件下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩,所得混合物直接通过反相快速色谱柱(C18柱) 进行纯化,在25分钟内用5%→60%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器,UV254纳米;得到化合物1-10b(白色油,200.0毫克,产率90%)。MS(ESI,m/z):613.3/615.2[M+H]
+。
步骤12’
在25摄氏度,氮气保护搅拌条件下,向化合物1-10b(200.0毫克,0.3毫摩尔,1.0当量)的乙腈/水(4/1,10毫升)溶液中分批加入高碘酸钠(431.8毫克,1.9毫摩尔,6.0当量)。反应液在25摄氏度条件下搅拌反应0.5小时,反应过程通过液质和薄层层析来监控。反应完全后,用二氯甲烷(30毫升 x 3)萃取。合并后有机层用30毫升饱和氯化钠溶液洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩,得到化合物1-11b(油,189.0毫克,产率100%)。MS(ESI,m/z):581.2/583.2[M+H]
+。
步骤13’
在25摄氏度,搅拌条件下,向化合物1-11b(189.0毫克,0.3毫摩尔,1.0当量)的甲醇(5.0毫升)溶液中加入化合物1-4(146.0毫克,0.3毫摩尔,1.2当量),然后在25摄氏度下加入冰乙酸(20.0毫克,0.3毫摩尔,1.2当量)和氰基硼氢化钠(37.0毫克,0.6毫摩尔,2.0当量),反应液在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩,所得混合物通过制备级高效液相色谱进行纯化,色谱柱XSelect CSH Prep C18 OBD,19 x 250毫米,5微米;流动相A:水(0.05%甲酸),流动相B:乙腈;流速:25毫升/分钟;在7分钟内用25%→55%的B相进行洗脱,检测器UV 220/235纳米,得到化合物1-12b(黄绿色固体,60.0毫克,产率22%)。MS(ESI,m/z):978.4/980.4[M+H]
+。
步骤14’
在室温搅拌条件下,向化合物1-12b(55.0毫克,0.04毫摩尔,1当量)的二氯甲烷(5.0毫升)溶液中加入三氟乙酸(2.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液 质和薄层层析来监控。反应完全后,减压浓缩除去多余的试剂,所得混合物通过制备级高效液相色谱进行纯化,色谱柱XSelect CSH Prep C18 OBD,19 x 250毫米,5微米;流动相A:水(0.05%甲酸),流动相B:乙腈;流速:25毫升/分钟;在7分钟内用10%→35%的B相进行洗脱,检测器UV 220/235纳米。得到化合物1b(黄绿色固体,20.0毫克,产率36%)。MS(ESI,m/z):878.4/880.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ11.10(s,1H),8.23–8.19(m,1H),7.96–7.93(m,1H),7.81(d,J=8.3Hz,1H),7.60–7.55(m,1H),7.47–7.41(m,1H),7.29(d,J=2.4Hz,1H),7.26–7.18(m,2H),7.12(d,J=8.6Hz,1H),7.08–7.05(m,1H),7.01(d,J=7.0Hz,1H),6.56(t,J=6.0Hz,1H),5.08–5.01(m,1H),4.36(t,J=6.5Hz,2H),3.84–3.79(m,4H),3.33–3.26(m,4H),3.01–2.94(m,4H),2.93–2.82(m,2H),2.63–2.52(m,2H),2.44–2.24(m,10H),2.06–1.98(m,1H),1.94–1.85(m,2H),1.61–1.42(m,4H)。
实施例2
4-((5-(4-(5-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)戊基)哌嗪-1-基)戊基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮二甲酸盐2
步骤1:
在25摄氏度搅拌条件下,将N-(5-溴戊基)邻苯二甲酰亚胺(5.0克,16.8毫摩尔,1.0当量)和1-叔丁氧羰基哌嗪(3.46克,1.86毫摩尔,1.1当量)溶于100毫升的乙腈中,然后加入N,N-二异丙基乙胺(4.36克,33.8毫摩尔,2.0当量)。将混合物升温至80摄氏度并在该温度下搅拌16小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品。所得粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物2-1(棕色油状物,6.6克,产率97%)。MS(ESI,m/z):402.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.86–7.80(m,2H),7.74–7.69(m,2H),3.71–3.67(m,2H),3.45–3.42(m,4H),2.40–2.33(m,6H),1.74–1.66(m,2H),1.61–1.52(m,2H),1.46(s,9H),1.40–1.33(m,2H)。
步骤2:
在25摄氏度搅拌条件下,将化合物2-1(3克,7.1毫摩尔,1.0当量)溶于100毫升乙醇中,然后加入水合肼(2.57克,41.1毫摩尔,5.5当量)。将反应体系升温至80摄氏度并在该温度下搅拌16小时,反应过程通过液质监控。反应结束后,将反应液冷却至25摄氏度,然后将反应液进行过滤,滤饼用20毫升乙醇冲洗三次,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→20%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物2-2(无色油状物,1.79克,产率88%)。MS(ESI,m/z):272.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ3.46–3.42(m,4H),2.71–2.68(m,2H),2.38–2.32(m,6H),1.55–1.30(m,17H)。
步骤3:
在25摄氏度氮气保护搅拌条件下,将2-(2,6氧代-哌啶-3-基)-4-氟基-异吲哚-1,3-二酮(700毫克,2.5毫摩尔,1.0当量)和化合物2-2(756.6克,2.8毫摩尔,1.1当量)溶于10毫升的N-甲基吡咯烷酮,然后加入N,N-二异丙基乙胺(655.1毫克,5.1毫摩尔,2.0当量)。将反应体系升温至90摄氏度并在该温度下反应2小时,反应过程通过液质监控。反应结束后,将反应液冷却至25摄氏度。反应液通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5→95%的乙腈/水流动相(0.1%甲酸)进行洗脱;检测器UV254纳米;得到化合物2-3(黄色油状物,502.3毫克,产率37%)。MS(ESI,m/z):528.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.29(s,1H),7.52–7.48(m,1H),7.10(d,J=7.1Hz,1H),6.88(d,J=8.5Hz,1H),6.24–6.22(m,1H),4.93–4.89(m,1H),3.60–3.59(m,4H),3.31–3.26(m,1H),2.91–2.59(m,15H),1.73–1.64(m,2H),1.46(s,9H)。
步骤4:
在25摄氏度搅拌条件下,将化合物2-3(200毫克,0.36毫摩尔,1.0当量)溶于3毫升的二氯 甲烷中,将体系降温至0摄氏度,然后向混合物中滴加1毫升三氟乙酸。加完反应体系升温至25摄氏度并在该温度下反应2小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品。得到的粗产品2-4(220毫克)直接用于下一步。MS(ESI,m/z):428.2[M+H]
+。
步骤5:
在25摄氏度氮气保护搅拌条件下,将5-己烯醇(5.0克,49.9毫摩尔,1.0当量)溶于500毫升的二氯甲烷中,然后加入(1,1,1-三乙酰氧基)-1,1-二氢-1,2-苯碘酰-3(1H)-酮(42.4克,99.8毫摩尔,2.0当量)。混合物在25摄氏度条件下反应2小时,反应过程通过薄层层析来监控。反应结束后,将反应液用饱和碳酸氢钠溶液(500毫升 x 2)萃取,合并有机相并用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法进行纯化(展开剂体系:石油醚/甲基叔丁基醚=20/1),得到化合物2-5(淡黄色油状物,700毫克,产率15%)。
1H NMR(400MHz,DMSO-d
6)δ9.67(t,J=1.5Hz,1H),5.83–5.73(m,1H),5.05–4.96(m,2H),2.45–2.41(m,2H),2.06–1.99(m,2H),1.65–1.58(m,2H)。
步骤6:
在25摄氏度氮气保护条件下,向500毫升反应瓶中依次加入3-(甲胺基)丙醇(10.0克,106.5毫摩尔,1.0当量),氯丙烯(9.4克,123.4毫摩尔,1.1当量),N,N-二异丙基乙胺(29.0克,224.371毫摩尔,2当量)和乙腈(100毫升)。所得混合物在80摄氏度反应16小时,反应过程通过液质和薄层色谱层析监控。反应结束后,反应液冷却至室温,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→12%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物2-6(黄色油状,9.1克,产率63%)。MS(ESI,m/z):130.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ6.10(s,1H),6.05–5.91(m,1H),5.42–5.36(m,2H),3.79–3.76(m,2H),3.42(d,J=6.9,2H),2.99–2.94(m,2H),2.56(s,3H),1.94–1.86(m,2H)。
步骤7:
在25摄氏度氮气保护条件下,依次向500毫升三口瓶中加入化合物1-7(12.0克,23.7毫摩尔,1.0 当量),化合物2-6(4.8克,35.6毫摩尔,1.50当量),氟化铯(7.2克,47.4毫摩尔,2.0当量);N,N-二异丙基乙胺(6.1克,47.4毫摩尔,2.0当量)和N-甲基吡咯烷酮(240毫升)。所得混合物在100摄氏度反应16小时,反应过程通过液质和薄色谱层层析监控。反应结束后,反应液冷却至室温,过滤,滤液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物2-7(黄色油状,4.5克,产率29%)。MS(ESI,m/z):572.1/574.1/576.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.71(d,J=1.96Hz,1H),6.02–5.97(m,1H),5.47–5.37(m,2H),4.53–4.50(m,2H),3.79–3.77(m,4H),3.66–3.64(m,4H),3.47(d,J=7.04Hz,2H),3.05–3.01(m,2H),2.58(s,3H),2.28–2.21(m,2H),1.50(s,9H)。
步骤8:
在25摄氏度氮气保护条件下,依次向250毫升三口瓶中加入化合物2-7(4.40克,7.3毫摩尔,1.0当量),4-(4,4,5,5-四甲基-1,3,2-二氧杂环芳烃-2-基)萘-2-醇(2.70g,9.485毫摩尔,1.30当量),碳酸钾(2.12克,14.6毫摩尔,2.0当量),甲烷磺酸(2-二环己基膦基-2',4',6'-三-异丙基-1,1'-联苯基)(2'-氨基-1,1'-联苯-2-基)钯(II)(650毫克,0.73毫摩尔,0.1当量),2-二-叔丁膦基-2',4',6'-三异丙基联苯(366.1毫克,0.73毫摩尔,0.1当量),80毫升1,4-二氧六环和20毫升水。所得混合物在60摄氏度反应3小时,反应过程通过液质和薄色谱层层析监控。反应结束后,反应液冷却至室温,减压除去多余的溶剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→12%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物2-8(白色固体,2.4克,产率49%)。MS(ESI,m/z):636.3/638.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.74–7.72(m,2H),7.43–7.39(m,1H),7.29–7.27(m,2H),7.22–7.19(m,1H),7.08(d,J=2.4Hz,1H),5.91–5.81(m,1H),5.20–5.12(m,2H),4.49–4.42(m,2H),3.81–3.80(m,4H),3.67–3.65(m,4H),3.06(d,J=6.64Hz,2H),2.63–2.59(m,2H),2.26(s,3H),2.09–2.02(m,2H),1.51(s,9H)。
步骤9:
通过制备级手性高效液相色谱法对化合物2-8(2.4克)进行手性拆分:手性柱CHIRALPAK IC,2 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/升氨),流动相B:乙醇;流速:20毫升/分钟;在25分钟内用50%的B相进行洗脱,检测器UV 220/254纳米,得到两个产品。较短保留时间(5.92 分钟)的产品为化合物2-8a(淡黄色油状液体,1.1克,回收率44%);较长保留时间(9.07分钟)的产品为化合物2-8b(淡黄色油状液体,0.95克,回收率37%)。
步骤10:
在25摄氏度氮气保护条件下,依次向50毫升三口瓶中加入化合物2-8a(1.1克,1.6毫摩尔,1.0当量),1,3-二甲基巴比妥酸(404.97毫克,2.4毫摩尔,1.5当量),四(三苯基膦)钯(99.9毫克,0.082毫摩尔,0.05当量)和二氯甲烷(20毫升),所得混合物在25摄氏度反应5小时,反应过程通过液质和薄色谱层层析监控。反应结束后减压浓缩除去多余的溶剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%甲酸)进行洗脱,检测器UV254纳米,得到化合物2-9a(淡黄色固体,970毫克,产率94%)。MS(ESI,m/z):596.2/598.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.61(s,1H),7.69–7.66(m,2H),7.35–7.31(m,1H),7.28–7.27(m,1H),7.18–7.08(m,3H),4.52–4.49(m,2H),3.82–3.64(m,8H),3.26(brs,2H),3.00–2.91(m,2H),2.50(s,3H),2.21–2.18(m,2H),1.49(s,9H)。
步骤11:
在25摄氏度搅拌条件下,将化合物2-9a(200.0毫克,0.336毫摩尔,1.0当量),醋酸(33.2毫克,0.53毫摩尔,1.5当量),氰基硼氢化钠(69.6毫克,1.05毫摩尔,3.0当量)溶于4毫升甲醇中,然后缓慢滴加化和物2-5(54.3毫克,0.53毫摩尔,1.5当量),所得混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质监控。反应结束后,反应液减压浓缩得到粗产品。所得粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物2-10a(白色固体,190毫克,产率76%)。MS(ESI,m/z):678.3/680.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.75–7.72(m,2H),7.42–7.38(t,J=7.4Hz,1H),7.31–7.30(m,1H),7.27–7.25(m,1H),7.21–7.14(m,1H),7.15(d,J=2.4Hz,1H),5.74–5.63(m,1H),4.97–4.90(m,2H),4.52–4.48(m,2H),3.87–3.84(m,4H),3.69–3.66(m,4H),2.99–2.94(m,2H),2.71–2.68(m,2H),2.52(s,3H),2.22–2.18(m,2H),2.06–196(m,2H),1.64–1.56(m,2H),1.51(s,9H),1.36–1.28(m,2H)。
步骤12:
在25摄氏度氮气保护搅拌条件下,将化合物2-10a(120毫克,0.17毫摩尔,1.0当量)和N-甲基吗啉氧化物(24.9毫克,0.21毫摩尔,1.2当量)溶于丙酮/水(4/1,5毫升)溶液中,然后在0摄氏度下缓慢加入锇酸钾(6.52毫克,0.017毫摩尔,0.1当量),反应液在0摄氏度搅拌条件下反应2小时,反应过程通过液质监控。反应结束后,反应液通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的乙腈/水(0.1%氨水)流动相进行洗脱,检测器UV254纳米,得到化合物2-11a(白色固体,80毫克,产率63%)。MS(ESI,m/z):712.3/714.3[M+H]
+。
步骤13:
在25摄氏度搅拌条件下,将化合物2-11a(80.0毫克,0.11毫摩尔,1.0当量)溶于乙腈/水(体积比4/1,5毫升)的混合溶剂中,然后加入高碘酸钠(120.1毫克,0.54毫摩尔,5.0当量)。反应液在25摄氏度搅拌条件下反应0.5小时,反应过程通过液质监控。反应结束后,将反应液倒入20毫升的水中进行稀释,再用二氯甲烷(15毫升 x 3)萃取,合并有机相,合并后有机相再用50毫升饱和食盐水洗涤,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品2-12a(80毫克,产率88%),直接用于下一步反应。MS(ESI,m/z):680.3/682.3[M+H]
+。
步骤14:
在25摄氏度氮气保护搅拌条件下,化合物2-4(64.8毫克)和氰基硼氢化钠(18.7毫克,0.28毫摩尔,3.0当量)溶于3毫升甲醇中,然后加入化合物2-12a(80.0毫克,0.09毫摩尔,1.0当量)。混合物在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,反应液通过制备级硅胶薄层层析法进行纯化(展开剂体系:二氯甲烷/甲醇=7/1),得到化合物2-13a(黄色油状物,90.0毫克,产率83%)。MS(ESI,m/z):1091.5/1093.5[M+H]
+。
步骤15:
在25摄氏度搅拌条件下,将化合物2-13a(90.0毫克,0.082毫摩尔,1.00当量)溶于3毫升的二氯甲烷中,将反应体系将至0摄氏度,然后滴加1毫升三氟乙酸。反应液在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,反应液减压浓缩得到粗产品,所得到的粗产品通过制备级高效液相色谱纯化,制备条件:反相柱XBridge Shield RP18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用8%-35%流动相B洗脱7分钟;检测器UV254/220纳米;得到化合物2(黄色固体,30毫克,产率33.7%)。MS(ESI,m/z):991.5/993.5[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.50(s,2H),8.03(d,J=1.6Hz,1H),7.79–7.76(m,1H),7.59–7.54(m,1H),7.46–7.41(m,1H),7.29(d,J=2.4Hz,1H),7.26–7.18(m,2H),7.08–7.05(m,3H),5.09–5.03(m,1H),4.62–4.58(m,2H),4.10–4.06(m,4H),3.39–3.37(m,5H),3.29(s,4H),3.13–3.08(m,2H),2.94–2.57(m,17H),2.32–2.23(m,2H),2.18–2.06(m,1H),1.82–1.36(m,12H);
19F NMR(282MHz,CD
3OD)δ-123.15。
实施例3
4-((5-(4-(5-((3-((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)戊基)哌嗪-1-基)戊基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮二甲酸盐3a;4-((5-(4-(5-((3-((R或S)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)戊基)哌嗪-1-基)戊基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮二三氟乙酸盐3b
步骤1
在25摄氏度搅拌条件下,向250毫升单口瓶中加入3-(甲氨基)-1-丙醇(3.0克,33.7毫摩尔,1.0当量)和乙腈(30.0毫升),然后加入6-氯-1-己烯(4.0克,33.7毫摩尔,1.0当量)和碳酸钾(9.3克,67.3毫摩尔,2.0当量)。所得混合物在70摄氏度条件下搅拌反应16个小时。反应过程通过液质 和薄层层析来监控。反应结束后,混合物冷却至25摄氏度,过滤,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,用0%→12%甲醇(2.5%氨甲醇)/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物3-1(黄色油状物,3.3克,产率54%)。MS(ESI,m/z):172.2[M+H]
+;
1HNMR(300MHz,CDCl
3)δ5.89–5.74(m,1H),5.07–4.93(m,2H),3.86–3.79(m,2H),2.65–2.59(m,2H),2.42–2.36(m,2H),2.26(s,3H),2.14–2.03(m,2H),1.77–1.67(m,2H),1.59–1.33(m,4H)。
步骤2
在0摄氏度,氮气保护搅拌条件下,向化合物1-6(6.00克,17.3毫摩尔,1.0当量)的超干二氯甲烷(70.0毫升)溶液中加入N,N-二异丙基乙胺(7.04克,51.7毫摩尔,3当量),随后滴加3,8-二氮杂双环[3.2.1]辛烷-8-羧酸叔丁酯(3.86克,17.3毫摩尔,1.0当量)的超干二氯甲烷(30.0毫升)溶液。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,冷却至室温。混合物通过减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用0%→50%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物3-2(黄色固体,8.1克,产率88%)。MS(ESI,m/z):505.0/507.1/509.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.75(d,J=2.0Hz,1H),4.52–4.29(m,4H),3.79–3.50(m,2H),2.04–1.89(m,2H),1.80–1.66(m,2H),1.52(s,9H)。
步骤3
在25摄氏度氮气保护条件下,向化合物3-2(2.0克,3.8毫摩尔,1.00当量)的N-甲基吡咯烷酮(20.0毫升)的溶液中,依次加入3-1(1.4克,7.5毫摩尔,2.0当量),氟化铯(1.2克,7.5毫摩尔,2.0当量)以及N-二异丙基乙胺(1.5克,11.2毫摩尔,3.0当量)。所得混合物在120摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至25摄氏度。反应液直接通过反相色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水(0.1%碳酸氢铵)流动相进行洗脱,检测器UV254/220纳米,得到化合物3-3(红色油状物,1.4克,产率52%)。MS(ESI,m/z):640.2/642.2/644.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.71(d,J=2.0Hz,1H),5.88–5.72(m, 1H),5.08–4.87(m,2H),4.56–4.46(m,2H),4.44–4.26(m,4H),3.68–3.48(m,2H),2.76–2.64(m,2H),2.56–2.44(m,2H),2.37(s,3H),2.18–2.03(m,4H),2.02–1.92(m,2H),1.84–1.73(m,2H),1.66–1.56(m,2H),1.53(s,9H),1.48–1.36(m,2H)。
步骤4
在25摄氏度氮气保护条件下,向100毫升单口瓶中依次加入化合物3-3(1.4克,2.2毫摩尔,1.0当量),4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(0.8克,2.8毫摩尔,1.3当量),氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(166.4毫克,0.2毫摩尔,0.1当量),磷酸钾(0.9克,4.1毫摩尔,2.0当量)以及四氢呋喃/水(10/1,15.0毫升)。所得混合物在60摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至25摄氏度,减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物3-4(红色油状物,1.2克,产率78%)。MS(ESI,m/z):704.3/706.3[M+H]
+;
1HNMR(400MHz,CDCl
3)δ7.75–7.67(m,2H),7.41–7.35(m,1H),7.32(d,J=2.4Hz,1H),7.26–7.23(m,1H),7.20–7.14(m,2H),5.73–5.62(m,1H),4.97–4.88(m,2H),4.52–4.41(m,4H),4.40–4.31(m,2H),3.75–3.47(m,2H),2.97–2.87(m,2H),2.68–2.57(m,2H),2.46(s,3H),2.26–2.15(m,2H),2.04–1.95(m,4H),1.86–1.75(m,2H),1.66–1.58(m,2H),1.53(s,9H),1.34–1.27(m,2H)。
步骤5
对步骤4中得到的化合物3-4(1.2克)进行手性拆分,拆分条件为:手性柱NB-Lux 5μm i-Cellulose-5,2.12 x 25厘米,5微米);流动相A:正己烷/二氯甲烷(5/1)(0.5%2摩尔每升氨甲醇),流动相B:乙醇;流速:20毫升/分钟;柱温:35摄氏度;用0%→20%流动相B洗脱20分钟;检测器UV220/254纳米;得到两个产品。保留时间较短(RT
1=6分钟)的化合物为3-4a(白色固体,520.0毫克,回收率43%),MS(ESI,m/z):704.3/706.3[M+H]
+;保留时间较长(RT
2=10.5分钟)的化合物为3-4b(白色固体,480.0毫克,回收率40%),MS(ESI,m/z):704.3/706.3[M+H]
+。
步骤6
在25摄氏度搅拌条件下,将3-4a(114.0毫克,0.14毫摩尔,1.0当量)溶于丙酮/水(4/1,2.0毫升)的混合溶液中,再加入锇酸钾(5.2毫克,0.02毫摩尔,0.1当量),然后在0摄氏度搅拌条件下,加入N-甲基吗啉氧化物(25.0毫克,0.2毫摩尔,1.5当量)。反应液在0摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液恢复至25摄氏度。反应液直接通过反相色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水(0.1%氨水)流动相进行洗脱;检测器UV254/220纳米;得到化合物3-5a(棕色油状物,92.0毫克,产率88%)。MS(ESI,m/z):738.4/740.4[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.75–7.67(m,2H),7.41–7.34(m,1H),7.29–7.27(m,1H),7.26–7.24(m,1H),7.21–7.15(m,1H),7.10–7.06(m,1H),4.73–4.65(m,1H),4.52–4.28(m,6H),3.66–3.57(m,2H),3.55–3.50(m,1H),3.38–3.30(m,1H),2.62–2.54(m,2H),2.40–2.36(m,2H),2.22(s,3H),2.04–1.77(m,6H),1.55–1.30(m,15H)。
步骤7
在25摄氏度搅拌条件下,将化合物3-5a(92.0毫克,0.14毫摩尔,1.0当量)溶于乙腈/水(4/1,4.0毫升)中,再加入高碘酸钠(181.7毫克,0.9毫摩尔,6.0当量)。所得混合物在25摄氏度条件下反应0.5小时,反应过程通过液质和薄层层析来监控。反应结束后,将反应液倒入10毫升水中,然后用二氯甲烷(10毫升 x 3)萃取。合并后有机相再用20毫升饱和食盐水洗涤。洗涤后有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品3-6a(105.1毫克),直接用于下一步。MS(ESI,m/z):706.4/708.4[M+H]
+。
步骤8
在25摄氏度搅拌条件下,向50毫升单口瓶中依次加入化合物2-4(80.6毫克,0.15毫摩尔,1.0当量),氰基硼氢化钠(29.5毫克,0.5毫摩尔,3.0当量)和甲醇(3.0毫升)。然后将化合物3-6a(105.1 毫克,0.2毫摩尔,1.0当量)溶于2.0毫升甲醇中,缓慢滴加至反应液中。所得混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品用高效液相纯化,制备条件:反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用17%→43%流动相B洗脱7分钟;检测器UV254/220纳米;得到产品3-7a(黄色油状物,110.0毫克,产率66%)。MS(ESI,m/z):1117.5/1119.5[M+H]
+。
步骤9
在25摄氏度搅拌条件下,将化合物3-7a(110.0毫克,0.1毫摩尔,1.0当量)溶于二氯甲烷(2.0毫升)中,在0摄氏度搅拌条件下,加入三氟乙酸(1.0毫升)。加完反应液恢复至25摄氏度,并在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品用高效液相纯化,制备条件:反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用5%→16%流动相B洗脱7分钟;检测器UV 254/220纳米;得到产品3a(黄色固体,27毫克,产率26%)。MS(ESI,m/z):1017.5/1019.5[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.52(s,2H),8.02–7.98(m,1H),7.81–7.73(m,1H),7.60–7.52(m,1H),7.47–7.39(m,1H),7.29(d,J=2.4Hz,1H),7.28–7.18(m,2H),7.09–7.03(m,3H),5.10–5.02(m,1H),4.70–4.54(m,4H),4.06–3.96(m,2H),3.82(d,J=13.4Hz,2H),3.40–3.35(m,2H),3.32–3.25(m,2H),3.13–3.04(m,2H),2.94–2.52(m,18H),2.33–2.20(m,2H),2.16–1.98(m,5H),1.84–1.55(m,8H),1.54–1.36(m,4H);
19F NMR(282MHz,CD
3OD)δ-123.18。
步骤6’
在25摄氏度搅拌条件下,将化合物3-4b(120.0毫克,0.17毫摩尔,1.0当量)溶于丙酮/水(4/1,2.0毫升)中,再加入锇酸钾(6.3毫克,0.02毫摩尔,0.1当量),然后在0摄氏度搅拌条件下,加入N-甲基吗啉氧化物(30.0毫克,0.3毫摩尔,1.5当量)。反应液在0摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液恢复至25摄氏度。反应液直接通过反相色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水(0.1%氨水)流动相进行洗脱;检测器UV254/220纳米;得到化合物3-5b(棕色油状物,100.0毫克,产率80%)。MS(ESI,m/z):738.4/740.4[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.74–7.68(m,2H),7.41–7.36(m,1H),7.29–7.24(m,2H),7.21 –7.15(m,1H),7.10–7.07(m,1H),4.54–4.38(m,4H),4.36–4.27(m,2H),3.71–3.56(m,2H),3.55–3.50(m,1H),3.49(s,1H),3.38–3.30(m,1H),2.63–2.52(m,2H),2.43–2.33(m,2H),2.23(s,3H),2.08–1.90(m,4H),1.87–1.74(m,2H),1.52(s,9H),1.50–1.42(m,2H),1.40–1.28(m,4H)。
步骤7’
在25摄氏度搅拌条件下,将化合物3-5b(100.0毫克,0.14毫摩尔,1.0当量)溶于乙腈/水(4:1,4.0毫升)中,再加入高碘酸钠(165.1毫克,0.8毫摩尔,6.0当量)。反应液在25摄氏度条件下反应0.5小时,反应过程通过液质和薄层层析来监控。反应结束后,将反应液倒入10毫升水中,然后用二氯甲烷(10毫升 x 3)萃取。合并后有机相再用20毫升饱和食盐水洗涤。洗涤后有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品3-6b(95.0毫克),直接用于下一步。MS(ESI,m/z):706.4/708.4[M+H]
+。
步骤8’
在25摄氏度搅拌条件下,向50毫升单口瓶中依次加入化合物2-4(72.8毫克,0.13毫摩尔,1.00当量),氰基硼氢化钠(25.4毫克,0.4毫摩尔,3.0当量)和甲醇(3.0毫升)。然后将化合物3-6b(95.0毫克,0.1毫摩尔,1.0当量)溶于2.0毫升甲醇中,缓慢滴加至反应液中。混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品用高效液相纯化,制备条件,反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米,流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用17%→43%流动相B洗脱7分钟;检测器UV254/220纳米;得到产品3-7b(黄色油状物,120.0毫克,产率76%)。MS(ESI,m/z):1117.5/1119.5[M+H]
+。
步骤9’
在25摄氏度搅拌条件下,将化合物3-7b(120.0毫克,0.1毫摩尔,1.0当量)溶于二氯甲烷(2.0 毫升)中,在0摄氏度搅拌条件下,加入三氟乙酸(1.0毫升)。加完反应液恢复至25摄氏度,并在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品用高效液相纯化,制备条件:反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用5%→16%流动相B洗脱7分钟;检测器UV 254/220纳米;得到产品3b(黄色固体,30.0毫克,产率23%)。MS(ESI,m/z):1017.5/1019.5[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.03–7.98(m,1H),7.81–7.75(m,1H),7.62–7.52(m,1H),7.47–7.39(m,1H),7.29(d,J=2.4Hz,1H),7.28–7.17(m,2H),7.10–7.03(m,3H),5.12–5.02(m,1H),4.74–4.54(m,4H),4.21–4.15(m,2H),3.94–3.83(m,2H),3.42–3.35(m,4H),3.22–3.13(m,2H),3.02–2.58(m,18H),2.36–2.23(m,2H),2.18–2.05(m,5H),1.86–1.56(m,8H),1.55–1.36(m,4H);
19F NMR(282MHz,CD
3OD)δ-123.07。
实施例4
4-((4-(4-(4-((3-((4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-((S或R)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮四三氟乙酸盐4a;4-((4-(4-(4-((3-((4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-((R或S)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮四三氟乙酸盐4b。
步骤1
在25摄氏度搅拌条件下,向反应瓶中依次加入3-(甲胺基)-1-丙醇(2.50克,26.6毫摩尔,1.00当量),醋酸(2.40克,40.0毫摩尔,1.50当量),氰基硼氢化钠(5.02克,79.9毫摩尔,3.00当量)和甲醇(30毫升)。然后向其中滴加4-戊烯醛(2.60克,29.4毫摩尔,1.10当量)。所得混合物在25摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→11%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物4-1(淡黄色油,4.4克,产率99%)。MS(ESI,m/z):158.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ5.84–5.72(m,1H),5.15–5.05(m,2H),3.84(t,J=5.5Hz,2H),3.20(t,J=6.8Hz,2H),3.05–2.97(m,2H),2.80(s,3H),2.25–2.14(m,2H),2.04–1.92(m,2H),1.91–1.78(m,2H)。
步骤2
在25摄氏度氮气保护条件下,向化合物3-2(2.0克,3.8毫摩尔,1.0当量)的N-甲基吡咯烷酮(20.0毫升)的溶液中,依次加入化合物4-1(1.2克,7.5毫摩尔,2.0当量),氟化铯(1.2克,7.5毫摩尔,2.0当量)和N-二异丙基乙胺(1.5克,11.3毫摩尔,3.0当量)。反应液在120摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至25摄氏度。过滤,滤液直接通过反相色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水(0.1%碳酸氢铵)流动相进行洗脱;检测器UV254/220纳米,得到化合物4-2(橘黄色油状物,1.0克,产率43%)。MS(ESI,m/z):626.2/628.1/630.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.69(d,J=2.0Hz,1H),5.86–5.72(m,1H),5.08–4.90(m,2H),4.48(t,J=6.4Hz,2H),4.45–4.25(m,4H),3.70–3.45(m,2H),2.69(t,J=7.2Hz,2H),2.50(t,J=7.6Hz,2H),2.36(s,3H),2.17–2.03(m,4H),2.00–1.92(m,2H),1.82–1.74(m,2H),1.70–1.60(m,2H),1.52(s,9H)。
步骤3
在25摄氏度氮气保护条件下,向100毫升单口瓶中依次加入化合物4-2(1.0克,1.5毫摩尔,1.0当量)和4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(646.3毫克,2.4毫摩尔,1.5当量),氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(125.5毫克,0.2毫摩尔,0.1当量),磷酸钾(677.1毫克,3.0毫摩尔,2.0当量)和四氢呋喃/水(10/1,11.0毫升)混合溶剂。所得混合物在60摄氏度条件下搅拌反应两小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至25摄氏度,减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→10%甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物4-3(红色油状物,800.0毫克,产率77%)。MS(ESI,m/z):690.3/692.3[M+H]
+;
1HNMR(400MHz,CDCl
3)δ7.78–7.70(m,2H),7.44–7.37(m,1H),7.32–7.26(m,2H),7.23–7.17(m,1H),7.13–7.10(m,1H),5.78–5.65(m,1H),5.01–4.88(m,2H),4.55–4.25(m,6H),3.76–3.45(m,2H),2.86–2.72(m,2H),2.62–2.48(m,2H),2.38(s,3H),2.20–2.08(m,2H),2.06–1.92(m,4H),1.86–1.76(m,2H),1.73–1.60(m,2H),1.53(s,9H)。
步骤4
对步骤3中得到的化合物4-3(800毫克)进行手性拆分,拆分条件为:手性柱CHIRALPAK IA,2 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/每升氨甲醇),流动相B:异丙醇;流速:20毫升/分钟;用30%流动相B洗脱22分钟;检测器UV220/210纳米;得到两个产品。保留时间较短(RT
1=10分钟)的化合物为4-3a(红色固体,330.0毫克,回收率41%),MS(ESI,m/z):690.3/692.3[M+H]
+;保留时间较长(RT
2=16分钟)的化合物为4-3b(红色固体,230.0毫克,回收率29%),MS(ESI,m/z):690.3/692.3[M+H]
+。
步骤5
在25摄氏度搅拌条件下,将化合物4-3a(200.0毫克,0.3毫摩尔,1.0当量)溶于丙酮/水(4/1,5.0毫升)中,再加入锇酸钾(10.7毫克,0.03毫摩尔,0.1当量),然后在0摄氏度搅拌条件下,加入N-甲基吗啉氧化物(50.9毫克,0.4毫摩尔,1.5当量)。所得混合物在0摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液恢复至25摄氏度。反应液直接通过反相色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水(0.1%氨水)流动相进行洗脱;检测器UV254/220纳米;得到化合物4-4a(红色固体,180.0毫克,产率86%)。MS(ESI,m/z):724.3/726.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.76–7.68(m,2H),7.42–7.36(m,1H),7.31–7.27(m,2H),7.22–7.16(m,1H),7.15–7.11(m,1H),4.56–4.26(m,6H),3.76–3.32(m,6H),2.74–2.62(m,1H),2.60–2.50(m,1H),2.48–2.34(m,2H),2.25(s,3H),2.10–1.93(m,4H),1.89–1.76(m,2H),1.72–1.60(m,3H),1.52(s,9H)。
步骤6
在25摄氏度搅拌条件下,将化合物4-4a(75.0毫克,0.1毫摩尔,1.0当量)溶于乙腈/水(4:1, 2.0毫升)中,再加入高碘酸钠(132.9毫克,0.6毫摩尔,6.0当量)。所得混合物在25摄氏度条件下反应0.5小时,反应过程通过液质和薄层层析来监控。反应结束后,将反应液倒入10毫升水中,然后用二氯甲烷(15毫升 x 3)萃取。合并后有机相用15毫升饱和食盐水洗涤。洗涤后有机相用无水硫酸钠干燥后,过滤,滤液减压浓缩得到粗产品4-5a(60.0毫克),直接用于下一步。MS(ESI,m/z):692.9/694.9[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向50毫升单口瓶中依次加入化合物1-4(35.8毫克,0.1毫摩尔,1.0当量),氰基硼氢化钠(16.3毫克,0.2毫摩尔,3.0当量)和甲醇(3.0毫升)。然后将化合物4-5a(60.0毫克,0.1毫摩尔,1.00当量)溶于2.0毫升甲醇中,缓慢滴加至反应液中。所得混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→10%甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物4-6a(黄色固体,62.0毫克,产率66%)。MS(ESI,m/z):1089.5/1090.5[M+H]
+。
步骤8
在25摄氏度搅拌条件下,将化合物4-6a(62.0毫克,0.1毫摩尔,1.0当量)溶于二氯甲烷(3.0毫升)中,在0摄氏度搅拌条件下,加入三氟乙酸(1.0毫升)。加完反应液恢复至25摄氏度,并在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品用高效液相纯化,制备条件:高压反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用25%-54%流动相B洗脱7分钟;检测器UV254/220纳米;得到产品4a(黄色固体,38.0毫克,产率46%)。MS(ESI,m/z):990.4/992.4[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.00(s,1H),7.78–7.73(m,1H),7.68–7.52(m,1H),7.45–7.38(m,1H),7.27(d,J=2.4Hz,1H),7.24–7.16(m,2H),7.08–7.01(m,3H),5.08–5.01(m,1H),4.76–4.68(m,2H),4.64–4.54(m,2H),4.25(s,2H),3.96–3.86(m,2H),3.50–3.34(m,4H),3.28–3.10(m,6H),3.09–2.97(m,5H),2.96–2.63(m,9H),2.35–2.24(m,2H),2.19–2.04(m,5H),1.88–1.62(m,8H);
19F NMR(377MHz,CD
3OD)δ-77.14,-123.09。
步骤5’
在25摄氏度搅拌条件下,将化合物4-3b(200.0毫克,0.3毫摩尔,1.0当量)溶于丙酮/水(4/1,5.0毫升)中,再加入锇酸钾(10.7毫克,0.03毫摩尔,0.1当量),然后在0摄氏度搅拌条件下,加入N-甲基吗啉氧化物(50.9毫克,0.4毫摩尔,1.5当量)。所得混合物在0摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液恢复至25摄氏度。反应液直接通过反相色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254/220纳米;得到化合物4-4b(红色固体,170.0毫克,产率81%)。MS(ESI,m/z):724.3/726.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.75–7.69(m,2H),7.42–7.35(m,1H),7.31–7.27(m,2H),7.22–7.15(m,1H),7.14–7.11(m,1H),4.54–4.28(m,6H),3.74–3.32(m,6H),2.74–2.61(m,1H),2.59–2.47(m,1H),2.46–2.32(m,2H),2.26–2.20(m,3H),2.08–1.92(m,4H),1.88–1.74(m,2H),1.70–1.59(m,3H),1.52(s,9H)。
步骤6’
在25摄氏度搅拌条件下,将化合物4-4b(80.0毫克,0.1毫摩尔,1.0当量)溶于乙腈/水(4:1,2.0毫升)中,再加入高碘酸钠(141.8毫克,0.6毫摩尔,6.0当量)。所得混合物在25摄氏度条件下反应0.5小时,反应过程通过液质和薄层层析来监控。反应结束后,将反应液倒入10毫升水中,然后用二氯甲烷(15毫升 x 3)萃取。合并后有机相用20毫升饱和食盐水洗涤,洗涤后有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品4-5b(70.0毫克),直接用于下一步。MS(ESI,m/z):692.9/694.9[M+H]
+。
步骤7’
在25摄氏度搅拌条件下,向50毫升单口瓶中依次加入化合物1-4(42.6毫克,0.1毫摩尔,1.0当量),氰基硼氢化钠(19.3毫克,0.2毫摩尔,3.0当量)和甲醇(3.0毫升)。然后将化合物4-5b(70.0 毫克,0.1毫摩尔,1.0当量)溶于2.0毫升甲醇中,缓慢滴加至反应液中。所得混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→10%甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物4-6b(黄色固体,73.0毫克,产率65%)。MS(ESI,m/z):1089.5/1090.5[M+H]
+。
步骤8’
在25摄氏度搅拌条件下,将化合物4-6b(73.0毫克,0.1毫摩尔,1.0当量)溶于二氯甲烷(3.0毫升)中,在0摄氏度搅拌条件下,加入三氟乙酸(1.0毫升)。加完反应恢复至25摄氏度,并在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品用高效液相纯化,制备条件:高压反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用25%→54%流动相B洗脱7分钟;检测器UV 254/220纳米;得到产品4b(黄色固体,52.0毫克,产率78%)。MS(ESI,m/z):990.4/992.4[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.02–7.98(m,1H),7.78–7.72(m,1H),7.59–7.52(m,1H),7.45–7.38(m,1H),7.27(d,J=2.4Hz,1H),7.24–7.16(m,2H),7.09–7.01(m,3H),5.09–5.01(m,1H),4.76–4.68(m,2H),4.64–4.54(m,2H),4.25(s,2H),3.95–3.86(m,2H),3.51–3.34(m,4H),3.28–3.17(m,5H),3.17–2.97(m,6H),2.92(s,3H),2.91–2.62(m,6H),2.34–2.24(m,2H),2.20–2.04(m,5H),1.88–1.64(m,8H);
19F NMR(377MHz,CD
3OD)δ-77.19,-123.09。
实施例5
4-((4-(4-((3-((4-((1S,4S)-2,5-二氮杂二环[2.2.2]辛烷-2-基)-6-氯-8-氟-7-((S或R)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮甲酸盐5a;4-((4-(4-((3-((4-((1S,4S)-2,5-二氮杂二环[2.2.2]辛烷-2-基)-6-氯-8-氟-7-((R或S)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮二甲酸盐5b
步骤1:
在25摄氏度氮气保护搅拌条件下,将7-溴-2,4,6-三氯-8-氟喹唑啉(780毫克,2.2毫摩尔,1.0当量)和三乙胺(716.8毫克,7.1毫摩尔,3.0当量)溶于15毫升1,4-二氧六环中,将体系降温至0摄氏度,然后加入(1S,4S)-2,5-二氮杂双环[2.2.2]辛烷-2-羧酸叔丁酯(501.2毫克,2.36毫摩尔,1.0当量)。混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化(流动相用0%→20%乙酸乙酯/二氯甲烷梯度洗脱),所得馏分通过减压旋蒸除去溶剂,得到化合物5-1(黄色固体,1.16克,产率97%)。MS(ESI,m/z):505.1/507.1/509.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.98–7.88(m,1H),5.17(s,1H),4.53–4.20(m,2H),4.05–3.92(m,1H),3.83–3.75(m,1H),3.67–3.52(m,1H),2.36–2.08(m,2H),2.02–1.74(m,2H),1.48(s,9H)。
步骤2:
在25摄氏度氮气保护搅拌条件下,将化合物5-1(1.1克,2.06毫摩尔,1.0当量)和3-[甲基(戊-4-烯-1-基)氨基]丙醇(683.5毫克,4.1毫摩尔,2.0当量)溶于10毫升的N-甲基吡咯烷酮中,然后加入N,N-二异丙基乙胺(842.6毫克,6.2毫摩尔,3.0当量)和氟化铯(660.2毫克,4.1毫摩尔,2.0当量)。加完将体系温度升至100摄氏度并在该温度下反应4小时,反应过程通过液质监控。反应结束后,将反应液冷却至25摄氏度,反应液通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→95%的甲醇/水(0.1氨水)流动相进行洗脱;检测器UV254纳米;得到化合物5-2(白色固体,670毫克,产率49%)。MS(ESI,m/z):626.2/628.2/630.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.87–7.83(m,1H),5.83–5.73(m,1H),5.08–4.95(m,3H),4.50–4.47(m,3H),4.31–4.21(m,1H),4.00–3.92(m,1H),3.81–3.77(m,1H),3.63–3.53(m,1H),2.79–2.76(m,2H),2.60–2.56(m,2H),2.42(s,3H),2.31–2.24(m,1H),2.17–2.06(m,5H),2.97–1.88(m,1H),1.86–1.76(d,J=10.5Hz,1H),1.74–1.66(m,2H),1.47(s,9H)。
步骤3:
在25摄氏度氮气保护搅拌条件下,向40毫升反应瓶中依次加入将化合物5-2(670毫克,1.02毫摩尔,1.0当量),4-(4,4,5,5-四甲基-1,3,2-二氧杂环芳烃-2-基)萘-2-醇(411.4毫克,1.6毫摩尔,1.5当量),氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(79.7毫克,0.1毫摩尔,0.1当量),磷酸钾(431.0毫克,2.03毫摩尔,2.0当量)和四氢呋喃/水(10/1,7.7毫升),所得混合物在60摄氏度反应3小时,反应过程通过液质监控。反应结束后,将反应液冷却至25摄氏度,减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化(流动相用0%→10%甲醇/二氯甲烷梯度洗脱),所得馏分通过减压旋蒸除去溶剂,得到化合物5-3(白色固体,480毫克,产率68%)。MS(ESI,m/z):690.3/692.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.89(d,J=11.6Hz,1H),7.72(d,J=8.3Hz,1H),7.43–7.36(m,1H),7.30–7.27(m,2H),7.21–7.16(m,1H),7.14–7.09(m,1H),5.84–5.66(m,1H),5.14–5.06(m,1H),5.02–4.86(m,2H),4.53–4.39(m,3H),4.36–4.30(m,1H),4.10–3.98(m,1H),3.81(d,J=11.9Hz,1H),3.68–3.53(m,1H),2.61(t,J=7.4Hz,2H),2.40(t,J=7.8Hz,2H),2.27(s,3H),2.14–1.79(m,9H),1.61–1.54(m,2H),1.51–1.46(m,9H)。
步骤4:
通过制备级手性高效液相色谱法对步骤3所得化合物5-3(480毫克)进行手性拆分:手性柱CHIRAL ART Amylose-SA,3 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/升氨),流动相B:异丙醇;流速:40毫升/分钟;在30分钟内用30%的B相进行洗脱,检测器UV 220/254纳米。得到两个产品,较短保留时间(10.17分钟)的产品为化合物5-3a(白色固体,180毫克,回收率38%),化合物5-3a:MS(ESI,m/z):690.3/692.3[M+H]
+;较长保留时间(17.32分钟)的产品为化合物5-3b(白色固体,160毫克,回收率33.3%),化合物5-3b:MS(ESI,m/z):690.3/692.3[M+H]
+。
步骤5:
在25摄氏度搅拌条件下,将化合物5-3a(180毫克,0.25毫摩尔,1.0当量)和N-甲基吗啉氧化物(43.5毫克,0.37毫摩尔,1.5当量)溶于丙酮/水(4/1,5毫升)中,然后在0摄氏度下缓慢加入锇酸钾(18.26毫克,0.05毫摩尔,0.2当量)。反应液在0摄氏度搅拌条件下反应3小时,反应过程通过液质监控。反应结束后,反应液通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水(0.1%氨水)流动相进行洗脱;检测器UV254纳米;得到化合物5-4a(白色固体,155毫克,产率82%)。MS(ESI,m/z):724.3/726.3[M+H]
+
步骤6:
在25摄氏度搅拌条件下,将化合物5-4a(75.0毫克,0.098毫摩尔,1.0当量)溶于乙腈/水(4/1,5毫升)中,然后加入高碘酸钠(132.9毫克,0.62毫摩尔,6.0当量)。反应液在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,将反应液倒入20毫升的水中,再用二氯甲烷(10毫升 x 3次)萃取,合并有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品5-5a(80毫克,产率93%)直接用于下一步反应。MS(ESI,m/z):692.3/694.3[M+H]
+。
步骤7:
在25摄氏度氮气保护搅拌条件下,将化合物1-4(49.8毫克,0.09毫摩尔,1.0当量)和氰基硼氢化钠(18.4毫克,0.28毫摩尔,3.0当量)溶于4毫升甲醇中,然后加入化合物5-5a(80.0毫克,0.09毫摩尔,1.0当量)。所得混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质监控。反应结束后,反应液通过制备级硅胶薄层层析法进行纯化(展开剂体系:二氯甲烷/甲醇=8/1),得到化合物5-6a(黄色油状物,80毫克,产率75%)。MS(ESI,m/z):1089.5/1091.5[M+H]
+。
步骤8:
在25摄氏度搅拌条件下,将化合物5-6a(80.0毫克,0.073毫摩尔,1.00当量)溶于3毫升二氯甲烷中,将反应体系将至0摄氏度,然后滴加1毫升三氟乙酸。反应液在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品。所得到的粗产品通过高效液相纯化,制备条件:高压反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用5%→20%流动相B洗脱10分钟;检测器UV254/220纳米;得到化合物5a(黄色固体,20毫克,产率25%)。MS(ESI,m/z):989.5/991.5[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.53(s,1H),8.15(d,J=1.7Hz,1H),7.77(d,J=8.3Hz,1H),7.57–7.52(m,1H),7.46–7.40(m,1H),7.29(d,J=2.4Hz,1H),7.25–7.18(m,2H),7.06–7.03(m,3H),5.09–5.01(m,2H),4.57–4.53(m,2H),4.45(d,J=11.9Hz,1H),4.29(d,J=11.5Hz,1H),3.72–3.66(m,2H),3.46–3.33(m,4H),3.19(t,J=7.6Hz,2H),3.00(t,J=7.6Hz,2H),2.88–2.39(m,18H),2.21–19.7(m,6H),1.75–1.61(m,8H);
19F NMR(282MHz,CD
3OD)δ-123.20。
步骤5’:
在25摄氏度搅拌条件下,将化合物5-3b(160毫克,0.22毫摩尔,1.0当量)和N-甲基吗啉氧化物(40.73毫克,0.35毫摩尔,1.5当量)溶于丙酮/水(4/1,5毫升)中,然后在0摄氏度下缓慢加入锇酸钾(17.1毫克,0.05毫摩尔,0.2当量)。反应液在0摄氏度搅拌条件下反应3小时,反应过程通过液质监控。反应结束后,反应液通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的甲醇/水(0.1%氨水)流动相进行洗脱;检测器UV254纳米;得到化合物5-4b(白色固体,145毫克,产率86%)。MS(ESI,m/z):724.3/726.3[M+H]
+
步骤6’:
在25摄氏度搅拌条件下,将化合物5-4b(75.0毫克,0.098毫摩尔,1.0当量)溶于乙腈/水(4/1,5毫升)中,然后加入高碘酸钠(132.9毫克,0.62毫摩尔,6.0当量)。反应液在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,将反应液倒入20毫升水中,二氯甲烷(10毫升 x 3)萃取,合并有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品5-5b(80毫克,产率93%)直接用于下一步反应。MS(ESI,m/z):692.3/694.3[M+H]
+。
步骤7’:
在25摄氏度氮气保护搅拌条件下,将化合物1-4(49.8毫克,0.09毫摩尔,1.0当量)和氰基硼氢化钠(18.4毫克,0.28毫摩尔,3.0当量)溶于4毫升甲醇中,然后加入化合物5-5b(80.0毫克,0.09毫摩尔,1.0当量)。混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质监控。反应结束后,反应液通过制备级硅胶薄层层析法进行纯化(展开剂体系:二氯甲烷/甲醇=8/1),得到化合物5-6b(黄色油状物,80毫克,产率75%)。MS(ESI,m/z):1089.5/1091.5[M+H]
+。
步骤8’:
在25摄氏度搅拌条件下,将化合物5-6b(75.0毫克,1.00当量)溶于3毫升二氯甲烷中,将反应体系将至0摄氏度,然后滴加1毫升三氟乙酸。反应液在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品。所得到的粗产品通过高效液相纯化,制备条件:高压反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.05%三氟乙酸),流动相B:甲醇;流速:25毫升/分钟;用27%→54%流动相B洗脱7分钟;检测器UV254/220纳米;得到化合物5b(黄色固体,12.6毫克)。MS(ESI,m/z):989.5/991.5[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.51(s,2H),8.15(d,J=1.6Hz,1H),7.77(d,J=8.3Hz,1H),7.59–7.51(m,1H),7.47–7.39(m,1H),7.29(d,J=2.4Hz,1H),7.26–7.15(m,2H),7.09–7.01(m,3H),5.11–5.00(m,2H),4.60–4.51(m,2H),4.51–4.39(m,1H),4.36–4.27(m,1H),3.75–3.65(m,2H),3.49–3.40(m,1H),3.38–3.35(m,1H),3.25–3.17(m,2H),3.07–2.98(m,2H),2.88–2.36(m,20H),2.27–2.03(m,6H),1.78–1.60(m,8H);
19F NMR(282MHz,CD
3OD)δ-123.64。
实施例6(合成方法I)
4-((6-(4-(6-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧 基)丙基)(甲基)氨基)己基)哌嗪-1-基)己基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮四三氟乙酸盐6
步骤1:
在25摄氏度氮气保护搅拌条件下,依次向250毫升单口瓶中加入N-(6-溴己基)酞亚酸(5.0克,16.1毫摩尔,1.0当量),1-叔丁氧羰基哌嗪(3.3克,17.7毫摩尔,1.1当量),乙腈(100毫升)和N,N-二异丙基乙胺(4.17克,32.2毫摩尔,2.0当量)。所得混合物在80摄氏度反应16小时。反应过程通过薄层层析来监控(R
f=0.7,二氯甲烷/甲醇=10/1)。反应结束后,将反应液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物6-1(棕色油状物,4.4克,产率69%)。MS(ESI,m/z):416.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.88–7.83(m,2H),7.76–7.71(m,2H),3.72–3.67(m,2H),3.46–3.43(m,4H),2.40–2.32(m,6H),1.72–1.36(m,17H)。
步骤2:
在25摄氏度搅拌条件下,将化合物6-1(4.4克,10.1毫摩尔,1.0当量)溶于100毫升乙醇中,加入水合肼(3.46克,55.3毫摩尔,5.5当量)。所得混合物体系在80摄氏度反应16小时,反应过程通过液质监控。反应结束后,将反应液冷却至0摄氏度,过滤除去不溶物,滤饼用150毫升乙醇洗涤三次,合并滤液,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→20%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物6-2(无色油状物,2.3克,产率76%)。MS(ESI,m/z):286.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ3.44–3.42(m,4H),2.70–2.67(m,2H),2.38–2.31(m,6H),1.53–1.3 0(m,17H)。
步骤3:
在25摄氏度氮气保护搅拌条件下,依次向100毫升单口瓶中加入6-2(1.1克,3.6毫摩尔,1.05当量),2-(2,6氧代-哌啶-3-基)-4-氟基-异吲哚-1,3-二酮(1克,3.4毫摩尔,1.0当量),N-甲基吡咯烷酮(10毫升)和入N,N-二异丙基乙胺(889毫克,6.9毫摩尔,2.0当量)。所得混合物升温至90摄氏度反应2小时,反应过程通过液质监控。反应结束后,将反应液冷却至25摄氏度。反应液通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→50%的甲醇/水(0.1%三氟乙酸)流动相进行洗脱;检测器UV254纳米;得到化合物6-3(黄色固体,360毫克,产率18%)。MS(ESI,m/z):542.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.25(s,1H),7.51–7.47(m,1H),7.09(d,J=7.1Hz,1H),6.88(d,J=8.5Hz,1H),6.24–6.21(m,1H),4.93–4.89(m,1H),3.46–3.43(m,4H),3.29–3.24(m,2H),2.91–2.69(m,3H),2.39–2.33(m,6H),2.16–2.01(m,1H),1.71–1.64(m,3H),1.54–1.25(m,14H)。
步骤4:
在0摄氏度搅拌条件下,依次向25毫升单口瓶中加入6-3(360毫克,0.6毫摩尔,1.0当量),二氯甲烷(3毫升)和三氟乙酸(1毫升)。所得混合物在25摄氏度反应1小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品,得到的粗产品6-4(350毫克,95%)直接用于下一步。MS(ESI,m/z):442.3[M+H]
+。
步骤5:
在0摄氏度搅拌条件下,将6-庚烯-1-醇(1.0克,8.3毫摩尔,1.0当量)溶于30毫升的二氯甲烷中,然后分批加入1,1,1-三乙酰氧基-1,1-二氢-1,2-苯碘氧基-3(1H)-酮(7.43克,17.5毫摩尔,2.0当量)。所得混合物在25摄氏度条件下反应2小时,反应过程通过薄层层析监控。反应结束后,用饱和亚硫酸氢钠/饱和硫代硫酸钠(30毫升/30毫升)将反应淬灭,混合物用二氯甲烷(50毫升 x 3)萃取,合并有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0% →20%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物6-5(无色油状物,680毫克,产率46%)。
1H NMR(400MHz,DMSO-d
6)δ9.67–9.66(m,1H),5.84–5.74(m,1H),5.04–4.93(m,2H),2.45–2.41(m,2H),2.05–1.99(m,2H),1.61–1.49(m,2H),1.39–1.31(m,2H)。
步骤6:
在25摄氏度搅拌条件下,将化合物2-9a(150.0毫克。0.24毫摩尔,1.0当量),醋酸(21.53毫克,0.36毫摩尔,1.5当量),氰基硼氢化钠(45.1毫克,0.72毫摩尔,3.0当量)溶于4毫升甲醇中,然后缓慢滴加化合物6-5(63.9毫克,0.36毫摩尔,1.5当量)。所得混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质监控。反应结束后,反应液通过制备级硅胶薄层层析法进行纯化(展开剂体系:二氯甲烷/甲醇=8/1),得到化合物6-6a(白色固体,150毫克,产率86%)。MS(ESI,m/z):692.3/694.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.73–7.69(m,2H),7.39–7.35(m,1H),7.30(d,J=2.4Hz,1H),7.24(d,J=8.3Hz,1H),7.18–7.14(m,2H),5.74–5.63(m,1H),4.95–4.87(m,2H),4.48–4.43(m,2H),3.86–3.84(m,4H),3.68–3.65(m,4H),3.01–2.97m,2H),2.69–2.65(m,2H),2.51(s,3H),2.19–2.15(m,2H),1.96–1.91(m,2H),1.60–1.54(m,2H),1.50(s,9H),1.32–1.25(m,2H),1.21–1.13(m,2H)。
步骤7:
在25摄氏度氮气保护搅拌条件下,将化合物6-6a(100毫克,0.137毫摩尔,1.0当量)和N-甲基吗啉氧化物(25.4毫克,0.21毫摩尔,1.5当量)溶于丙酮/水(4/1,5毫升)中,然后在0摄氏度下缓慢加入锇酸钾(5.3毫克,0.014毫摩尔,0.1当量)。所得混合物在0摄氏度搅拌条件下反应5小时,反应过程通过液质监控。反应结束后,反应液通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水(0.1%氨水)流动相进行洗脱;检测器UV254纳米;得到化合物6-7a(黄色油状物,77毫克,产率73%)。MS(ESI,m/z):726.3/728.3[M+H]
+。
步骤8:
在25摄氏度搅拌条件下,将化合物6-7a(75.0毫克,0.098毫摩尔,1.0当量)溶于乙腈/水(4/1,5毫升)中,然后加入高碘酸钠(132.5毫克,0.62毫摩尔,6.0当量)。混合物在25摄氏度反应1小时,反应过程通过液质监控。反应结束后,将反应液倒入10毫升的水中,用二氯甲烷(10毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品6-8a(70毫克,产率92%)直接用于下一步反应。MS(ESI,m/z):694.3/696.3[M+H]
+。
步骤9:
在25摄氏度氮气保护搅拌条件下,将化合物6-4(51.5毫克,0.09毫摩尔,1.0当量),氰基硼氢化钠(18.0毫克,0.27毫摩尔,3.0当量)溶于3毫升甲醇中,然后加入化合物6-8a(70.0毫克,0.09毫摩尔,1.0当量)。所得混合物在25摄氏度反应1小时,反应过程通过液质监控。反应结束后,反应液直接通过制备级硅胶薄层层析法进行纯化(展开剂体系:二氯甲烷/甲醇=6/1),得到化合物6-9a(黄色油状物,60毫克,产率52%)。MS(ESI,m/z):1119.5/1121.5[M+H]
+。
步骤10:
在0摄氏度搅拌条件下,向25毫升单口瓶中依次加入6-9a(50.0毫克,0.042毫摩尔,1.00当量),二氯甲烷(3毫升)和三氟乙酸(1毫升)。加完在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品。所得粗产品通过高效液相纯化,制备条件:高压反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;用5%→40%流动相B洗脱7分钟;检测器UV254/220纳米;得到化合物6(黄色固体,20毫克,产率31%)。MS(ESI,m/z):1019.5/1021.5[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.07(d,J=1.7Hz,1H),7.79(d,J=8.3Hz,1H),7.62–7.53(m,1H),7.49–7.41(m,1H),7.30(d,J=2.4Hz,1H),7.26–7.21(m,2H),7.11–7.03(m,3H),5.14–5.02(m,1H),4.70–4.56(m,2H),4.24–4.08(m,4H),3.56–3.35(m,11H),3.32–2.64(m,17H),2.41–2.25(m,2H),2.20–2.05(m,1H),1.84–1.61(m,8H),1.60–1.38(m,8H)。
实施例7
4-((7-(4-(7-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)庚基)哌嗪-1-基)庚基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮四三氟乙酸盐7
步骤1
在25摄氏度搅拌条件下,依次向100毫升圆底烧瓶中加入邻苯二甲酰亚胺钾盐(1.79克,9.181毫摩尔,1.0当量),丙酮(30.0毫升,387.661毫摩尔,42.23当量)和1,7-二溴庚烷(4.99克,18.36毫摩尔,2.0当量)。所得混合物在60摄氏度下反应16小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→40%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物7-1(无色油状物,2.3克,产率77%)。MS(ESI,m/z):324.0/326.0[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.87–7.82(m,2H),7.74–7.69(m,2H),3.70–3.66(m,2H),3.41–3.38(m,2H),1.88–1.81(m,2H),1.72–1.65(m,2H),1.47–1.34(m,6H)。
步骤2
在25摄氏度搅拌条件下,向100毫升圆底烧瓶中依次加入7-1(2.3克,6.739毫摩尔,1.0当量),1-叔丁氧羰基哌嗪(1.45克,7.413毫摩尔,1.1当量),乙腈(30.0毫升,542.204毫摩尔,80.45当量)和N,N-二异丙基乙胺(1.83克,13.478毫摩尔,2.00当量)。加完升温至80摄氏度下搅拌反应4小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩得到粗产品。 粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物7-2(亮黄色油状物,2.5克,产率86%)。MS(ESI,m/z):430.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.86–7.82(m,2H),7.73–7.69(m,2H),3.69–3.66(m,2H),3.44–3.41(m,4H),2.37–2.29(m,6H),1.71–1.64(m,2H),1.51–1.43(m,11H),1.36–1.27(m,6H)。
步骤3
在25摄氏度搅拌条件下,向500毫升圆底烧瓶中依次加入化合物7-2(2.5克,5.529毫摩尔,1.0当量),乙醇(100.0毫升)和水合肼(80%)(1.17克,22.116毫摩尔,4.00当量)。所得混合物在80摄氏度反应16小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物7-3(无色油状物,1.2克,产率72%)。MS(ESI,m/z):300.6[M+H]
+;
1H NMR(400MHz,CD
3OD)δ3.44–3.41(m,4H),2.64–2.61(m,2H),2.42–2.34(m,6H),1.56–1.45(m,13H),1.37–1.32(m,6H)。
步骤4
在25摄氏度搅拌条件下,向40毫升反应瓶中加入2-(2,6-二氧代-哌啶-3-基)-4-氟基-异吲哚-1,3-二酮(500.0毫克,1.720毫摩尔,1.0当量),N-甲基吡咯烷酮(5.00毫升),N,N-二异丙基乙胺(467.89毫克,3.44毫摩尔,2.0当量)和化合物7-3(569.16毫克,1.806毫摩尔,1.05当量)。加完升温至90摄氏度,并在此温度下反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物7-4(黄色固体,200毫克,产率20%)。MS(ESI,m/z):556.1[M+H]
+。
步骤5:
在0摄氏度搅拌条件下,向50毫升圆底烧瓶中依次加入化合物7-4(200.0毫克,0.342毫摩尔,1.0当量),二氯甲烷(6.00毫升)和三氟乙酸(2.00毫升)。所得混合物在25摄氏度反应1小时。反应过程通过液质来监控。反应结束后,减压浓缩得到化合物7-5粗产品(黄色固体,120毫克),该粗产品直接用于下一步。MS(ESI,m/z):456.8[M+H]
+。
步骤6
在25摄氏度搅拌条件下,将7-辛烯-1-醇(2.0克,14.819毫摩尔,1.0当量)溶于二氯甲烷(200.00毫升),然后向其中分批加入1,1,1-三乙酰氧基-1,1-二氢-1,2-苯碘氧基-3(1H)-酮(13.23克,29.638毫摩尔,2.00当量)。所得混合物在25摄氏度搅拌条件下反应2小时。反应过程通过薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→30%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物7-6(亮黄色油状物,1.5克,产率80%)。
1H NMR(400MHz,DMSO-d
6)δ9.66–9.65(m,1H),5.84–5.74(m,1H),5.03–4.92(m,2H),2.44–2.40(m,2H),2.04–1.98(m,2H),1.56–1.49(m,2H),1.39–1.22(m,4H)。
步骤7
在25摄氏度搅拌条件下,向8毫升反应瓶中依次加入化合物2-9a(120.0毫克,0.191毫摩尔,1.0当量),甲醇(2.00毫升),醋酸(17.23毫克,0.287毫摩尔,1.50当量)和氰基硼氢化钠(36.05毫克,0.573毫摩尔,3.00当量),最后加入化合物7-6(38.11毫克,0.287毫摩尔,1.50当量)。所得混合物在25摄氏度下反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物7-7a(棕色固体,110毫克,产率81%)。MS(ESI,m/z):706.2/708.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.74–7.72(m,2H),7.43–7.39(m,1H),7.29–7.27(m,2H),7.23–7.19(m,1H),7.07(d,J=2.4Hz,1H),5.82–5.71(m,1H),4.99–4.88(m,2H),4.50–4.47(m,2H),3.82–3.80(m,4H),3.68–3.66(m,4H),2.61–2.57(m,2H),2.40–2.36(m,2H), 2.27(s,3H),2.08–1.96(m,4H),1.51(s,9H),1.48–1.25(m,8H)。
步骤8
在0摄氏度搅拌条件下,向50毫升圆底烧瓶中依次加入化合物7-7a(99.00毫克,0.133毫摩尔,1.00当量),丙酮(3.20毫升)/水(0.80毫升),二水合锇酸钾(5.16毫克,0.013毫摩尔,0.10当量)和N-甲基吗啉氧化物(24.63毫克,0.2毫摩尔,1.50当量)。所得混合物在0摄氏度下反应2小时。反应过程通过液质监控。反应完全后,将反应液升温至25摄氏度,反应液浓缩后通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→95%的乙腈/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物7-8a(黄色油状,80毫克,产率81%)。MS(ESI,m/z):740.5/742.5[M+H]
+。
步骤9
在25摄氏度搅拌条件下,向50毫升圆底烧瓶中加入化合物7-8a(80.0毫克,0.103毫摩尔,1.0当量),乙腈(4.00毫升)/水(1.00毫升),然后向其中分批次加入高碘酸钠(138.68毫克,0.618毫摩尔,6.0当量)。所得混合物在25摄氏度下反应1小时。反应过程通过液质和薄层层析监控。反应结束后,将反应液倒入10毫升水中,用二氯甲烷(10毫升 x 3)萃取。合并后有机相再用20毫升饱和食盐水洗涤,洗涤后有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到7-9a粗产品(黄色固体,60毫克,产率82%),该粗产品克直接用于下一步。MS(ESI,m/z):708.3/710.3[M+H]
+。
步骤10
在25摄氏度搅拌条件下,向50毫升圆底烧瓶中加入化合物7-5(51.46毫克,0.107毫摩尔,1.00当量),氰基硼氢化钠(21.29毫克,0.321毫摩尔,3.00当量)和甲醇(3毫升),最后向上述反应体系中缓慢加入化合物7-9a(80.0毫克,0.107毫摩尔,1.0当量)。所得混合物在25摄氏度下反应2小时。反应过程通过液质和薄层层析来监控,反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析 法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物7-10a(黄色固体,80.0毫克,产率39%)。MS(ESI,m/z):1147.6/1149.6[M+H]
+。
步骤11
在0摄氏度搅拌条件下,向25毫升单口瓶中依次加入化合物7-10a(80.0毫克,0.042毫摩尔,1.00当量),二氯甲烷(3毫升)和三氟乙酸(1毫升)。反应液在25摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,将反应液减压浓缩得到粗产品。所得粗产品通过高效液相纯化,制备条件:高压反相柱XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;用5%→40%流动相B洗脱10分钟;检测器UV254/220纳米;得到化合物7(黄色固体,61.2毫克,产率37%)。MS(ESI,m/z):1047.5/1049.5[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.04(d,J=1.6Hz,1H),7.77–7.75(m,1H),7.56–7.52(m,1H),7.44–7.40(m,1H),7.28(d,J=2.4Hz,1H),7.22–7.17(m,2H),7.05–7.03(m,3H),5.07–5.02(m,1H),4.62–4.58(m,2H),4.14–4.12(m,4H),3.52–3.34(m,12H),3.29–2.64(m,16H),2.30–2.28(m,2H),2.12–2.06(m,1H),1.74–1.66(m,8H),1.43–1.29(m,12H);
19F NMR(377MHz,CD
3OD)δ-77.18,-123.05。
实施例8
4-(4-(4-(4-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁氧基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮二甲酸盐8
步骤1
在25摄氏度搅拌条件下,向化合物1-叔丁氧羰基哌嗪(5.0克,25.5毫摩尔,1.0当量)和4-氯-1-丁醇(3.5克,0.03毫摩尔,1.2当量)的乙腈(50.0毫升)溶液中加入碳酸氢钠(9.0克,102.0毫摩尔,4.0当量)和碘化钠(2.0克,12.8毫摩尔,0.5当量)。所得混合物在90摄氏度下反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液冷却至室温,减压浓缩得到粗产品。向粗产品中加入100.0毫升水,用乙酸乙酯(100毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物8-1(黄色液体,2.6克,产率36%)。MS(ESI,m/z):259.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ3.60–3.54(m,2H),3.50–3.42(m,4H),2.50–2.38(m,6H),1.74–1.65(m,4H),1.46(s,9H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向化合物8-1(819.0毫克,3.2毫摩尔,2.2当量)的四氢呋喃(1.0毫升)溶液中滴加双三甲基硅基胺基锂(1摩尔/升四氢呋喃溶液)(3.2毫升,3.2毫摩尔,2.2当量)。加完所得混合物在60摄氏度下继续搅拌反应1小时,然后向反应液中滴加2-(2,6-二氧代-哌啶-3-基)-4-氟-异吲哚-1,3-二酮(398.0毫克,1.4毫摩尔,1.0当量)的甲苯(6.0毫升)溶液。所得混合物在100摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物8-2(白色固体,80毫克,产率11%)。MS(ESI,m/z):515.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.24(s,1H),7.72–7.67(m,1H),7.46(d,J=7.2Hz,1H),7.21(d,J=8.2Hz,1H),4.99–4.93(m,1H),4.28–4.17(m,2H),3.52–3.40(m,4H),2.95–2.72(m,3H),2.52–2.36(m,4H),2.17–2.12(m,1H),1.99–1.89(m,2H),1.81–1.65(m,4H),1.45(s,9H)。
步骤3
在25摄氏度搅拌条件下,向化合物8-2(110.0毫克,0.20毫摩尔,1.0当量)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通 过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品8-3(灰白色固体,150.0毫克)。MS(ESI,m/z):415.4[M+H]
+。
步骤4
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物2-9a(200.0毫克,0.32毫摩尔,1.0当量),醋酸(28.7毫克,0.48毫摩尔,1.5当量)氰基硼氢化钠(60.0毫克,0.96毫摩尔,3.0当量)和甲醇(3.0毫升),然后向其中滴加4-戊烯醛(40.2毫克,0.48毫摩尔,1.5当量)的甲醇(0.5毫升)溶液。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物8-4a(白色固体,150.0毫克,产率71%)。MS(ESI,m/z):664.4/666.4[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.76–7.70(m,2H),7.43–7.38(m,1H),7.32(d,J=2.4Hz,1H),7.25–7.24(m,1H),7.22–7.16(m,1H),7.14(d,J=2.4Hz,1H),5.79–5.57(m,1H),5.01–4.92(m,2H),4.53–4.43(m,2H),3.90–3.76(m,4H),3.69–3.64(m,4H),2.96(s,2H),2.69(s,2H),2.52(s,3H),2.21(s,2H),2.06–1.98(m,2H),1.78–1.69(m,2H),1.51(s,9H)。
步骤5
在0摄氏度搅拌条件下,向化合物8-4a(65.0毫克,0.09毫摩尔,1.0当量)的丙酮/水(4/1,2.0毫升)溶液中加入N-甲基吗啉氧化物(16.3毫克,0.14毫摩尔,1.5当量)和锇酸钾二水合物(6.9毫克,0.02毫摩尔,0.2当量)。所得混合物在0摄氏度下继续搅拌反应5小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用30%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物8-5a(淡黄色固体,66.0毫克,产率96%)。MS(ESI,m/z):698.4/700.3[M+H]
+。
步骤6
在25摄氏度搅拌条件下,向化合物8-5a(50.0毫克,0.07毫摩尔,1.0当量)的乙腈/水(4/1,1.0毫升)溶液中加入高碘酸钠(91.9毫克,0.4毫摩尔,6.0当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入水(10.0毫升),二氯甲烷(10.0毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品8-6a(淡黄色油状液体,55.0毫克),粗品未进一步纯化,直接用于下一步反应。MS(ESI,m/z):666.2/668.2[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物8-3(36.6毫克,0.07毫摩尔,1.0当量)的甲醇(2.0毫升)溶液中加入氰基硼氢化钠(13.1毫克,0.20毫摩尔,3.0当量),然后滴加化合物8-6a(55.0毫克,0.07毫摩尔,1.0当量)的甲醇(0.5毫升)溶液。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物8-7a(无色油状液体,50.0毫克,产率67%)。MS(ESI,m/z):1064.4/1066.4[M+H]
+。
步骤8
在25摄氏度搅拌条件下,向化合物8-7a(65.0毫克,0.06毫摩尔,1.0当量)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在10分钟内用5%→19%B相梯度洗脱;检测器UV 254/220纳米;得到化合物8(白色固体,15.0毫克,产率24%)。MS(ESI,m/z):964.4/966.4 [M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ11.09(s,1H),8.25–8.14(m,2H),,7.97(s,1H),7.80(s,2H),7.54–7.40(m,3H),7.29(d,J=2.4Hz,1H),7.25–7.18(m,2H),7.07(d,J=2.4Hz,1H),5.11–5.05(m,1H),4.43–4.36(s,2H),4.20(t,J=6.2Hz,2H),3.89–3.81(m,5H),3.10–2.97(m,4H),2.93–2.87(m,1H),2.75–2.72(m,1H),2.63–2.60(m,1H),2.43–2.14(m,18H),2.09–1.86(m,4H),1.80–1.70(m,2H),1.63–1.54(m,2H),1.46–1.33(m,4H);
19F NMR(282MHz,DMSO-d
6)δ-122.27。
实施例9
4-((3-(4-(3-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丙基)哌嗪-1-基)丙基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮二甲酸盐9a;4-((3-(4-(3-((3-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丙基)哌嗪-1-基)丙基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮甲酸盐9b
步骤1
在25摄氏度搅拌条件下,向化合物3-(甲胺基)-1-丙醇(5.0克,56.1毫摩尔,1.0当量),二氯甲烷(50.0毫升)和N,N-二异丙基乙胺(21.7克,168.3毫摩尔,3.0当量)的混合液中滴加二碳酸二叔丁酯(15.9克,72.9毫摩尔,1.3当量)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→40%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物9-1(无色油状液体,10.1克,产率95%)。MS(ESI,m/z):190.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ3.57(s,2H),3.40(t,J=6.2Hz,2H),2.85(s,3H),1.78–1.64(m,2H),1.48(s,9H)。
步骤2
在25摄氏度搅拌条件下,向化合物9-1(3.0克,15.9毫摩尔,1.0当量)的二氯甲烷(30.0毫升)溶液中加入1,1,1-三乙酰氧基-1,1-二氢-1,2-苯碘氧基-3(1H)-酮(13.44克,31.8毫摩尔,2.0当量)。 所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→30%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物9-2(无色油状液体,1.4克,产率47%)。MS(ESI,m/z):188.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ9.83(t,J=1.7Hz,1H),3.57(t,J=6.6Hz,2H),2.89(s,3H),2.70(m,2H),1.47(s,9H)。
步骤3
在25摄氏度,氮气保护搅拌条件下,向50毫升三口烧瓶中加入4-(3-氨基丙基)哌嗪-1-羧酸叔丁酯(1.0克,4.1毫摩尔,1.0当量),N-甲基吡咯烷酮(10.0毫升),2-(2,6-二氧代-哌啶-3-基)-4-氟-异吲哚-1,3-二酮(1.1克,4.1毫摩尔,1.0当量)和N,N-二异丙基乙胺(0.94克,7.2毫摩尔,1.8当量)。所得混合物在90摄氏度下继续搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相色谱(C18)进行纯化,在35分钟内,用5%→95%的乙腈/水(0.1%三氟乙酸)流动相进行洗脱,检测器UV 254/220纳米。得到化合物9-3(黄色固体,650.0毫克,产率32%)。MS(ESI,m/z):500.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.23(s,1H),7.53–7.48(m,1H),7.12–7.10(m,1H),6.94(d,J=8.5Hz,1H),6.60(t,J=5.6Hz,1H),4.96–4.90(m,1H),3.52–3.49(m,4H),3.42–3.36(m,2H),2.97–2.68(m,3H),2.50(t,J=6.6Hz,2H),2.45–2.41(m,4H),2.23–2.04(m,1H),1.89–1.83(m,2H),1.48(s,9H)。
步骤4
在25摄氏度搅拌条件下,向化合物9-3(600.0毫克,1.1毫摩尔,1.0当量)的二氯甲烷(6.0毫升)溶液中加入三氟乙酸(1.5毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品9-4(黄色固体,700.0毫克)。该化合物不需进一步纯化,直接用于下一步合成。MS(ESI,m/z):400.1[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向化合物9-4(600.0毫克,1.1毫摩尔,1.0当量),的甲醇(7.0毫升)溶液中依次加入化合物9-2(355.3毫克,1.8毫摩尔,1.2当量)和氰基硼氢化钠(198.7毫克,3.0毫摩尔,2.0当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相色谱(C18)进行纯化,在20分钟内,用5%→70%的乙腈/水(0.1%三氟乙酸)流动相进行洗脱,检测器UV 254/220纳米,得到化合物9-5(黄色固体,435毫克,产率90%)。MS(ESI,m/z):571.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ10.63(s,1H),7.55–7.50(m,2H),7.13(d,J=7.0Hz,1H),6.90(d,J=8.6Hz,1H),4.99–4.93(m,1H),3.70–3.12(m,8H),3.07–2.51(m,14H),2.20–1.92(m,3H),1.86(s,2H),1.49(s,9H)。
步骤6
在25摄氏度搅拌条件下,向化合物9-5(400.0毫克,0.7毫摩尔,1.0当量)的二氯甲烷(9.0毫升)溶液中加入三氟乙酸(3.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品9-6(黄色固体,400.0毫克)。该化合物不需进一步纯化,直接用于下一步合成。MS(ESI,m/z):471.2[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物9-6(116.6毫克,0.2毫摩尔,1.2当量),醋酸钠(25.4毫克,0.3毫摩尔,1.5当量),醋酸(12.4毫克,0.2毫摩尔,1.0当量),氰基硼氢化钠(38.9毫克,0.6毫摩尔,3.0当量)和甲醇(2.0毫升),加完向其中化合物1-11a(114.0毫克,0.2毫摩尔,1.0当量)的甲醇(0.5毫升)溶液。所得混合物在25摄氏度下继续搅拌反应2小时,反应过 程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物9-7a(淡黄色固体,100.0毫克,产率49%)。MS(ESI,m/z):1035.4/1037.4[M+H]
+。
步骤8
在25摄氏度搅拌条件下,向化合物9-7a(50.0毫克,0.05毫摩尔,1.0当量)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(0.5毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→25%B相梯度洗脱;检测器UV 254/220纳米;得到化合物9a(黄色固体,25毫克,产率50%)。MS(ESI,m/z):935.4/937.4[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ11.11(s,1H),8.19(s,2H),7.98(d,J=1.7Hz,1H),7.81(d,J=8.2Hz,1H),7.59–7.53(m,1H),7.46–7.41(m,1H),7.29(d,J=2.4Hz,1H),7.21(d,J=5.0Hz,2H),7.15–6.97(m,3H),6.75(d,J=5.9Hz,1H),5.07–5.03(m,1H),4.39–4.34(m,2H),3.88–3.86(m,5H),3.37–3.30(m,3H),3.11–3.08(m,4H),2.95–2.81(m,2H),2.64–2.54(m,4H),2.32–2.28(m,15H),2.10–1.83(m,4H),1.75–1.54(m,4H)。
步骤7’
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物9-6(116.6毫克,0.2毫摩尔,1.2当量),醋酸钠(25.4毫克,0.3毫摩尔,1.5当量),醋酸(12.4毫克,0.2毫摩尔,1.0当量),氰基硼氢化钠(38.9毫克,0.6毫摩尔,3.0当量)和甲醇(2.0毫升),然后向其中加入化合物1-11b(114毫克,0.2毫摩尔,1.0当量)。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物9-7b(淡黄色固体,106毫克,产率52%)。MS(ESI,m/z):1035.4/1037.4[M+H]
+。
步骤8’
在25摄氏度搅拌条件下,向化合物9-7b(100.0毫克)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(0.5毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→25%B相梯度洗脱;检测器UV 254/220纳米;得到化合物9b(黄色固体,55毫克,产率58%)。MS(ESI,m/z):935.4/937.4[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.53(s,1H),8.00(s,1H),7.75–7.71(m,1H),7.54–7.50(m,1H),7.40–7.38(m,1H),7.30–7.16(m,3H),7.08–7.00(m,3H),5.04–4.87(m,1H),4.58–4.53(m,2H),4.04–4.01(m,4H),3.38(d,J=6.2Hz,2H),3.32–3.24(m,4H),3.13–3.05(m,2H),2.99–2.94(m,2H),2.89–2.65(m,12H),2.65–2.43(m,6H),2.21–2.17(m,2H),2.12–2.08(m,1H),1.94–1.88(m,2H),1.82–1.79(m,2H)。
实施例10
4-((4-(4-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮二甲酸盐10a;4-((4-(4-((3-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮二甲酸盐10b
步骤1
在25摄氏度,氮气保护搅拌条件下,向化合物1-7(1.23克,2.536毫摩尔,1.00当量)的N-甲基吡咯烷酮(50毫升)溶液中加入氟化铯(0.81克,5.072毫摩尔,2.0当量),化合物4-1(0.63克, 3.804毫摩尔,1.5当量)和N,N-二异丙基乙胺(0.69克,5.072毫摩尔,2.0当量)。所得混合物在100摄氏度下搅拌反应16小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物10-1(白色固体,760毫克,产率48%)。MS(ESI,m/z):600.3/602.3/604.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.68(d,J=2.0Hz,1H),5.81–5.68(m,1H),5.07–4.96(m,2H),4.51(t,J=6.0Hz,2H),3.86–3.71(m,4H),3.69–3.55(m,4H),3.13–3.01(m,2H),2.87–2.76(m,2H),2.64(s,3H),2.34–2.19(m,2H),2.15–2.07(m,2H),1.87–1.76(m,2H),1.48(s,9H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向化合物10-1(710.00毫克,1.142毫摩尔,1.00当量,96.7%)的四氢呋喃/水(10/1,11毫升)的溶液中依次加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(487.30毫克,1.714毫摩尔,1.5当量),磷酸钾(510.54毫克,2.285毫摩尔,2.0当量)和氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(94.50毫克,0.114毫摩尔,0.1当量)。所得混合物在60摄氏度条件下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物10-2(黄色固体,750毫克,产率98%)。MS(ESI,m/z):664.3/666.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.76–7.70(m,2H),7.44–7.38(m,1H),7.32–7.28(m,1H),7.28–7.26(m,1H),7.23–7.18(m,1H),7.10(d,J=2.5Hz,1H),5.80–5.66(m,1H),5.05–4.87(m,2H),4.55–4.44(m,2H),3.88–3.61(m,8H),2.82–2.68(m,2H),2.60–2.46(m,2H),2.44–2.32(m,3H),2.18–2.08(m,2H),2.06–1.99(m,2H),1.71–1.62(m,2H),1.51(s,9H)。
步骤3
通过制备级手性高效液相色谱法对化合物10-2(710毫克)进行手性拆分:手性柱CHIRAL ART Cellulose-SC,2 x 25厘米,5微米;流动相A:正己烷/二氯甲烷=5/1,流动相B:乙醇;流速:20毫 升/分钟;在3.3分钟内用50%的B相进行洗脱,检测器UV 220/230纳米。得到两个产品,较短保留时间(6.0分钟)的产品为化合物10-2a(黄绿色固体,300毫克,回收率43%),MS(ESI,m/z):664.3/666.3[M+H]
+;较长保留时间(7.4分钟)的产品为化合物10-2b(黄绿色固体,210毫克,回收率30%),MS(ESI,m/z):664.3/666.3[M+H]
+。
步骤4
在0摄氏度搅拌条件下,向化合物10-2a(300.00毫克,0.452毫摩尔,1.00当量)的水/四氢呋喃(1/4,10毫升)溶液中加入N-甲基吗啉氧化物(113.4毫克,0.9毫摩尔,2当量)和四氧化锇(6.0毫克,0.024毫摩尔,0.05当量)。所得混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物10-3a(黄绿色固体,240毫克,产率73%)MS(ESI,m/z):698.2/670.2[M+H]
+;
1H NMR(400MHz,CD
3OD)δ7.98(d,J=1.6Hz,1H),7.76–7.72(m,1H),7.43–7.37(m,1H),7.25(d,J=2.5Hz,1H),7.23–7.17(m,2H),7.03(d,J=2.4Hz,1H),4.54–4.46(m,2H),4.00–3.92(m,4H),3.74–3.65(m,4H),3.59–3.51(m,1H),3.41(d,J=5.5Hz,2H),2.67–2.59(m,2H),2.48–2.41(m,2H),2.29(s,3H),2.07–1.98(m,2H),1.75–1.55(m,4H),1.50(s,9H)1.39–1.27,(m,2H)。
步骤5
在0摄氏度搅拌条件下,向化合物10-3a(80.00毫克,0.115毫摩尔,1.00当量)的水/四氢呋喃(1/4,5毫升)溶液中加入高碘酸钠(147.04毫克,0.687毫摩尔,6.0当量)。所得混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入水(50毫升),用二氯甲烷(50毫升 x 3)萃取。合并后有机相用无水硫酸钠干燥,过滤,滤液浓缩得粗产品10-4a。MS(ESI,m/z):666.3/668.3[M+H]
+。
步骤6
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物1-4(71.06毫克,0.172毫摩尔,1.50当量),乙酸(13.76毫克,0.229毫摩尔,2.0当量),氰基硼氢化钠(10.80毫克,0.172毫摩尔,1.5当量,95%)和甲醇(3毫升),然后向其中滴加上述10-4a的甲醇(1毫升)溶液。所得混合物在25摄氏度下搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,混合物通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度:7分钟内用35%→61%B相梯度洗脱;检测器UV 254/220纳米;得到化合物10-5a(黄绿色固体,50毫克,产率41%)。MS(ESI,m/z):1063.3/1065.4[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物10-5a(50.00毫克,0.047毫摩尔,1.00当量)的二氯甲烷(2毫升)溶液中加入三氟乙酸(1毫升)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度:7分钟内用35%→60%B相梯度洗脱;检测器UV 254/220纳米;得到10a(黄绿色固体,15毫克,产率32%)。MS(ESI,m/z):963.5/965.5[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.35(s,2H),7.91(d,J=1.6Hz,1H),7.65(d,J=8.3Hz,1H),7.48–7.39(m,1H),7.36–7.27(m,1H),7.19–7.07(m,3H),6.99–6.88(m,3H),4.99–4.90(m,1H),4.48(t,J=5.8Hz,2H),4.03–3.92(m,4H),3.35–3.26(m,4H),3.26–3.22(m,4H),3.05–2.96(m,2H),2.79–2.46(m,18H),2.20–2.11(m,2H),2.05–1.93(m,1H),1.70–1.50(m,8H)。
步骤4’
在0摄氏度搅拌条件下,向化合物10-2b(210.00毫克,0.316毫摩尔,1.00当量)的水/四氢呋喃(1/4,10毫升)溶液中加入N-甲基吗啉氧化物(77.97毫克,0.632毫摩尔,2.0当量)和四氧化锇(4.23毫克,0.016毫摩尔,0.05当量)。所得混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物10-3b(黄绿色固体,150毫克,产率65%)。MS(ESI,m/z):698.2/700.2[M+H]
+;
1H NMR(400MHz,CD
3OD)δ7.99(d,J=1.7Hz,1H),7.74(d,J=8.3Hz,1H),7.44–7.37(m,1H),7.25(d,J=2.4Hz,1H),7.23–7.17(m,2H),7.03(d,J=2.4Hz,1H),4.50(t,J=6.3Hz,2H),3.98–3.90(m,4H),3.75–3.64(m,4H),3.60–3.52(m,1H),3.44–3.39(m,2H),2.69–2.60(m,2H),2.52–2.40(m,2H),2.34–2.28(m,3H),2.09–1.99(m,2H),1.76–1.54(m,4H),1.50(s,9H),1.39–1.27(m,2H)。
步骤5’
在0摄氏度搅拌条件下,向化合物10-3b(80.00毫克,0.099毫摩尔,1.00当量)的水/四氢呋喃(1/4,5毫升)溶液中加入高碘酸钠(133.82毫克,0.594毫摩尔,6.0当量)。所得混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入水(50毫升),用二氯甲烷萃取(50毫升 x 3)。合并后有机相用无水硫酸钠干燥,过滤,滤液浓缩得粗产物10-4b。MS(ESI,m/z):666.3/668.3[M+H]
+。
步骤6’
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物1-4(64.67毫克,0.149毫摩尔,1.5当量),乙酸(12.52毫克,0.198毫摩尔,2.0当量),氰基硼氢化钠(9.83毫克,0.149毫摩尔,1.5当量)和甲醇(2毫升),然后向其中滴加化合物10-4b的甲醇(1毫升)溶液。所得混合物在25摄氏度下搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,混合物通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度:7分钟内用35%→61%B相梯度洗脱;检 测器UV 254/220纳米;得到10-5b(黄绿色固体,50毫克,产率47%)。MS(ESI,m/z):1063.5/1065.4[M+H]
+。
步骤7’
在25摄氏度搅拌条件下,向化合物10-5b((50.00毫克,0.047毫摩尔,1.00当量)的二氯甲烷(2毫升)溶液中加入三氟乙酸(1毫升)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度:7分钟内用5%→25%B相梯度洗脱;检测器UV 254/220纳米;得到10b(黄绿色固体,17.6毫克,产率38%)。MS(ESI,m/z):963.4/965.5[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.35(s,2H),7.92(d,J=1.6Hz,1H),7.66(d,J=8.3Hz,1H),7.48–7.40(m,1H),7.35–7.27(m,1H),7.17(d,J=2.4Hz,1H),7.15–7.06(m,2H),6.97–6.90(m,3H),5.01–4.88(m,1H),4.49(t,J=5.9Hz,2H),4.08–3.92(m,4H),3.36–3.28(m,4H),3.27–3.22(m,4H),3.07–2.98(m,2H),2.88–2.39(m,18H),2.22–2.10(m,2H),2.05–1.94(m,1H),1.74–1.45(m,8H)。
实施例11
5-((4-(4-(4-((3-(((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮三氟乙酸盐11
步骤1
在25摄氏度,氮气保护搅拌条件下,向化合物1-2(3.5克,12.9毫摩尔,1.1当量)和2-(2,6-二氧代-哌啶-3-基)-4-氟-异吲哚-1,3-二酮(3.4克,11.7毫摩尔,1.0当量)的N-甲基吡咯烷酮(34.0毫升)溶液中加入N,N-二异丙基乙胺(3.2克,23.4毫摩尔,2.0当量)。所得混合物在90摄氏度下继续 搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液冷却至室温,反应液通过反相色谱(C18)进行纯化,在25分钟内,用5%→95%的乙腈/水(0.1%三氟乙酸)流动相进行洗脱,检测器:UV 254/220纳米。得到化合物11-1(黄绿色油状体,1.4克,产率22%)。MS(ESI,m/z):514.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.87(s,1H),7.62(d,J=8.3Hz,1H),7.02(d,J=2.1Hz,1H),6.75–6.71(m,1H),5.16(s,1H),4.97–4.91(m,1H),3.52–3.44(m,4H),3.24(t,J=6.3Hz,2H),2.98–2.59(m,3H),2.46(d,J=5.5Hz,6H),2.21–2.09(m,1H),1.82–1.59(m,4H),1.48(s,9H)。
步骤2
在25摄氏度搅拌条件下,向化合物11-1(350.0毫克,粗品)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品11-2(黄色固体,350毫克)。该化合物不需进一步纯化,直接用于下一步合成。MS(ESI,m/z):414.1[M+H]
+。
步骤3
在25摄氏度搅拌条件下,向反应瓶中加入化合物11-2(37.4毫克,0.07毫摩尔,1.2当量),醋酸钠(6.2毫克,0.07毫摩尔,1.0当量),醋酸(4.5毫克,0.07毫摩尔,1.0当量),氰基硼氢化钠(9.4毫克,0.14毫摩尔,2.0当量)和甲醇(2.0毫升),然后向其中滴加化合物10-4a(50.0毫克,0.07毫摩尔,1.0当量)的甲醇(0.5毫升)溶液。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物11-3a(黄色固体,35毫克,产率44%)。MS(ESI,m/z):1063.4/1065.4[M+H]
+。
步骤4
在25摄氏度搅拌条件下,向化合物11-3a(35.0毫克,0.03毫摩尔,1.0当量)的二氯甲烷(0.9毫升)溶液中加入三氟乙酸(0.3毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进 行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.05%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→24%B相梯度洗脱;检测器UV 254/220纳米;得到化合物11(黄色固体,15.0毫克,产率44%)。MS(ESI,m/z):963.4/965.4[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ11.07(s,1H),8.18(s,1H),8.01(d,J=1.6Hz,1H),7.82(d,J=8.3Hz,1H),7.56(d,J=8.3Hz,1H),7.47–7.42(m,1H),7.30(d,J=2.3Hz,1H),7.26–7.16(m,2H),7.14–7.11(m,1H),7.07(d,J=2.4Hz,1H),6.94(d,J=2.0Hz,1H),6.86–6.82(m,1H),5.06–5.00(m,1H),4.43–4.38(m,2H),3.91–3.89(m,6H),3.17–3.15(m,7H),2.94–2.80(m,1H),2.75–2.64(m,2H),2.64–2.55(m,2H),2.41–2.22(m,14H),2.02–1.96(m,3H),1.53–1.45(m,8H);
19F NMR(377MHz,DMSO-d
6)δ-73.48,-122.22。
实施例12
4-((4-(4-((3-((4-((1S,4S)-2,5-二氮杂二环[2.2.1]庚烷-2-基)-6-氯-8-氟-7-((S或R)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮甲酸盐12a;4-((4-(4-((3-((4-((1S,4S)-2,5-二氮杂二环[2.2.1]庚烷-2-基)-6-氯-8-氟-7-((R或S)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮二甲酸盐12b
步骤1
在0摄氏度,氮气保护搅拌条件下,向化合物1-6(1.5克,4.3毫摩尔,1.0当量)的超干1,4-二氧六环(15.0毫升)溶液中,加入三乙胺(1.8毫升,13.0毫摩尔,3.0当量),然后滴加(1S,4S)-2,5-二氮杂二环[2.2.1]庚烷-2-羧酸叔丁酯(0.9克,4.3毫摩尔,1.0当量)的超干1,4-二氧六环(3.0毫升)溶液。所得混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→35%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物12-1(黄色固体,1.9克,产率85%)。MS(ESI,m/z): 491.0/493.0/495.0[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.88(d,J=11.6Hz,1H),5.47–5.33(m,1H),4.86–4.66(m,1H),4.18–4.15(m,1H),4.05–3.77(m,1H),3.60–3.56(m,2H),2.08–2.05(m,2H),1.45(s,9H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向化合物12-1(1.5克,2.3毫摩尔,1.0当量)的N-甲基吡咯烷酮(15.0毫升)溶液中,依次加入氟化铯(0.9克,5.8毫摩尔,2.0当量),N,N-二异丙基乙胺(1.5毫升,8.7毫摩尔,3.0当量)以及化合物4-1(960.0毫克,5.8毫摩尔,2.0当量)。加完所得混合物升温至130摄氏度继续搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用50%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物12-2(淡黄色固体,750.0毫克,产率42%)。MS(ESI,m/z):612.2/614.2/616.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.80(d,J=11.6Hz,1H),5.88–5.75(m,1H),5.37–5.24(m,1H),5.07–4.90(m,2H),4.81–4.61(m,1H),4.50–4.46(m,2H),4.15–4.12(m,1H),3.99–3.47(m,3H),2.61–2.57(m,2H),2.48–2.36(m,2H),2.29(s,3H),2.14–2.03(m,6H),1.65–1.55(m,2H),1.47–1.44(m,9H)。
步骤3
在25摄氏度,氮气保护搅拌条件下,向化合物12-2(750.0毫克,1.2毫摩尔,1.0当量)的四氢呋喃/水(10/1,8.3毫升)溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(495.8毫克,1.7毫摩尔,1.5当量),磷酸钾(519.4毫克,2.3毫摩尔,2.0当量)和氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(96.2毫克,0.1毫摩尔,0.1当量)。所得混合物在60摄氏度条件下搅拌反应1.5小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物12-3(淡黄色固体,673.0毫克,产率 86%)。MS(ESI,m/z):676.4/678.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.88–7.85(m,1H),7.74–7.71(m,1H),7.42–7.38(m,1H),7.29(s,1H),7.28–7.16(m,2H),7.13–7.09(m,1H),5.83–5.69(m,1H),5.37–5.29(m,1H),5.05–4.87(m,2H),4.72–4.70(m,1H),4.48(s,2H),4.22–3.97(m,2H),3.80–3.78(m,1H),3.60–3.57(m,1H),2.69–2.64(m,2H),2.52–2.38(m,2H),2.31(s,3H),2.07–2.00(m,6H),1.67–1.57(m,2H),1.45(s,9H)。
步骤4
通过制备级手性高效液相色谱法对步骤3所得化合物12-3(673.0毫克)进行手性拆分:手性柱CHIRALPAK IA,2 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/升氨-甲醇),流动相B:异丙醇;流速:20毫升/分钟;在22分钟内用50%的B相进行洗脱;检测器UV 220/254纳米,得到两个产品。较短保留时间(4.75分钟)的产品为化合物12-3a(淡黄色固体,330.0毫克,回收率49%),化合物12-3a:MS(ESI,m/z):676.4/678.4[M+H]
+;较长保留时间(10.76分钟)的产品为化合物12-3b(淡黄色固体,260.0毫克,回收率39%),化合物12-3b:MS(ESI,m/z):676.4/678.4[M+H]
+。
步骤5
在0摄氏度搅拌条件下,向化合物12-3a(330.0毫克,0.5毫摩尔,1.0当量)的丙酮/水(4/1,6.3毫升)溶液中依次加入N-甲基吗啉氧化物(85.8毫克,0.7毫摩尔,1.5当量)和锇酸钾二水合物(18.0毫克,0.05毫摩尔,0.1当量)。所得混合物在0摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用50%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物12-4a(灰色固体,184毫克,产率56%)。MS(ESI,m/z):710.4/712.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.84(s,1H),7.75–7.72(m,1H),7.43–7.38(m,1H),7.31(s,2H),7.25–7.16(m,2H),5.36–5.32(m,1H),4.78–4.66(m,1H),4.48–4.45(m,2H),4.19–4.16(m,1H),4.04–4.01(m,1H),3.88–3.73(m,1H),3.67–3.49(m,4H),3.44–3.38(m,1H),2.70–2.54(m,2H),2.42(s,2H),2.26(s,3H),2.08–2.03(m,4H),1.68–1.65(m,3H),1.49(s,9H)。
步骤6
在25摄氏度搅拌条件下,向化合物12-4a(75.0毫克,0.1毫摩尔,1.0当量)的乙腈/水(4/1,1.5毫升)溶液中加入高碘酸钠(128.7毫克,0.6毫摩尔,6.0当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入水(10毫升),二氯甲烷(10毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品12-5a(淡黄色油状液体,80.0毫克),粗品直接用于下一步反应。MS(ESI,m/z):678.3/680.3[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物1-4(61.6毫克,0.14毫摩尔,1.2当量),醋酸钠(10.2毫克,0.12毫摩尔,1.0当量),醋酸(7.5毫克,0.12毫摩尔,1.0当量),氰基硼氢化钠(15.6毫克,0.24毫摩尔,2.0当量)和甲醇(2.0毫升),然后向其中滴加化合物12-5a(80.0毫克,0.12毫摩尔,1.0当量)的甲醇(0.5毫升)溶液。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物12-6a(黄色固体,46毫克,产率45%)。MS(ESI,m/z):1075.4/1077.4[M+H]
+。
步骤8
在25摄氏度搅拌条件下,向化合物12-6a(46.0毫克,0.04毫摩尔,1.0当量)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(0.5毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用10%→95%的甲醇/水流动相(0.1%甲酸)进行洗脱;检测器UV254纳米;得到化合物12a(黄色固体,20毫克,产率47%)。MS(ESI,m/z):975.4/977.4[M+H]
+;
1H NMR (300MHz,DMSO-d
6)δ11.11(s,1H),8.19(s,1H),8.05(d,J=1.7Hz,1H),7.82(d,J=8.3Hz,1H),7.57–7.55(m,1H),7.46–7.44(m,1H),7.30(d,J=2.4Hz,1H),7.24–7.21(m,1H),7.18–7.07(m,3H),7.02(d,J=7.0Hz,1H),6.57–6.53(m,1H),5.19(s,1H),5.06–5.02(m,1H),4.47–4.32(m,4H),4.27(s,2H),3.99(d,J=10.0Hz,2H),3.44–3.19(m,5H),2.92–2.80(m,1H),2.70–2.56(m,4H),2.37–2.34(m,6H),2.30–2.24(m,8H),2.12–2.10(m,1H),2.08–1.87(m,4H),1.50–1.44(m,8H);
19F NMR(282MHz,DMSO-d
6)δ-122.09。
步骤5’
在0摄氏度搅拌条件下,向化合物12-3b(260.0毫克,0.4毫摩尔,1.0当量)的丙酮/水(4/1,5.0毫升)溶液中依次加入N-甲基吗啉氧化物(67.6毫克,0.5毫摩尔,1.5当量)和锇酸钾二水合物(14.2毫克,0.04毫摩尔,0.1当量)。所得混合物在0摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用50%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物12-4b(灰色固体,200.0毫克,产率77%)。MS(ESI,m/z):710.4/712.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.84(s,1H),7.75–7.72(m,1H),7.43–7.38(m,1H),7.31(s,2H),7.25–7.16(m,2H),5.36–5.32(m,1H),4.78–4.66(m,1H),4.48–4.45(m,2H),4.19–4.16(m,1H),4.04–4.01(m,1H),3.88–3.73(m,1H),3.67–3.49(m,4H),3.44–3.38(m,1H),2.70–2.54(m,2H),2.42(s,2H),2.26(s,3H),2.08–2.03(m,4H),1.68–1.65(m,3H),1.49(s,9H)。
步骤6’
在25摄氏度搅拌条件下,向化合物12-4b(75.0毫克,0.1毫摩尔,1.0当量)的乙腈/水(4/1,1.5毫升)溶液中加入高碘酸钠(128.7毫克,0.6毫摩尔,6.0当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入水(10.0毫升),二氯甲烷(10.0毫升 x 3)萃取,合并有机相,有机相再用无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品12-5b(淡黄色油状液体,80.0毫克),粗品直接用于下一步反应。MS(ESI,m/z):678.4/680.4 [M+H]
+。
步骤7’
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物1-4(65.5毫克,0.12毫摩尔,1.2当量),醋酸钠(10.8毫克,0.13毫摩尔,1.0当量),醋酸(7.9毫克,0.13毫摩尔,1.0当量),氰基硼氢化钠(16.6毫克,0.25毫摩尔,2.0当量)和甲醇(2.0毫升)中,然后向其中滴加12-5b(85.0毫克,0.13毫摩尔,1.0当量)的甲醇(0.5毫升)溶液。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物12-6b(黄色固体,56毫克,产率49%)。MS(ESI,m/z):1075.4/1077.4[M+H]
+。
步骤8’
在25摄氏度搅拌条件下,向化合物12-6b(56.0毫克,0.05毫摩尔,1.0当量)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(0.5毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→95%的甲醇/水流动相(0.1%甲酸)进行洗脱;检测器UV254纳米;得到化合物12b(黄色固体,30毫克,产率57%)。MS(ESI,m/z):975.4/977.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ11.10(s,1H),8.19(s,2H),8.07–8.02(m,1H),7.81(d,J=8.3Hz,1H),7.58–7.54(m,1H),7.46–7.41(m,1H),7.29(d,J=2.4Hz,1H),7.24–7.13(m,2H),7.10(d,J=8.6Hz,1H),7.06–6.98(m,2H),6.55(t,J=5.9Hz,1H),5.20–5.17(m,1H),5.08–5.02(m,1H),4.38–4.32(m,4H),4.16(s,2H),3.93(d,J=9.9Hz,2H),3.31–3.25(m,3H),3.22–3.20(m,1H),2.94–2.83(m,1H),2.62–2.60(m,1H),2.58–2.55(m,3H),2.45–2.42(m,3H),2.39–2.31(m,4H),2.29–2.21(m,8H),2.10–2.00(m,2H),2.09–2.01(m,3H),1.61–1.50(m,2H),1.49–1.36(m,6H);
19F NMR(377MHz,DMSO-d
6)δ-122.02。
实施例13
4-((2-(2-(4-(2-(2-((3-((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)乙氧基)乙基)哌嗪-1-基)乙氧基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮五三氟乙酸盐13a;4-((2-(2-(4-(2-(2-((3- ((R或S)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)乙氧基)乙基)哌嗪-1-基)乙氧基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮五三氟乙酸盐13b。
步骤1
在25摄氏度搅拌条件下,向250毫升圆底烧瓶中加入邻苯二甲酰亚胺钾盐(5.0克,25.6毫摩尔,1.0当量),丙酮(50.0毫升)和2,2'-二溴二乙醚(11.9克,51.3毫摩尔,2.0当量)。将所得混合物升温至60摄氏度并在该温度下搅拌16小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用10%→30%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物13-1(类白色固体,6.4克,产率84%)。MS(ESI,m/z):298.0/300.0[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.90–7.82(m,2H),7.76–7.69(m,2H),3.94-3.90(m,2H),3.82-3.76(m,4H),3.41(t,J=6.2Hz,2H)。
步骤2
在25摄氏度搅拌条件下,向500毫升圆底烧瓶中加入化合物13-1(6.4克,20.4毫摩尔,1.0当量),1-叔丁氧羰基哌嗪(4.2克,22.4毫摩尔,1.1当量),乙腈(100.0毫升)和N,N-二异丙基乙胺(7.1毫升,40.8毫摩尔,2.0当量)。将混合物升温至80摄氏度并在该温度下搅拌4小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物13-2(深黄色油状液体,7.0克,产率85%)。MS(ESI,m/z):404.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.87–7.81(m,2H),7.76–7.68(m,2H),3.90(t,J=5.7Hz,2H),3.71(t,J=5.6Hz,2H),3.61(t,J=5.5Hz,2H),3.41–3.29(m,4H),2.53(t,J=5.5Hz,2H),2.37(t,J=5.1Hz,4H),1.45(s,9H)。
步骤3
在25摄氏度搅拌条件下,向化合物13-2(7.0克,16.5毫摩尔,1.0当量)的乙醇(300.0毫升)溶液中加入水合肼(4.1克,66.0毫摩尔,4.0当量)。所得混合物在80摄氏度下搅拌反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液冷却至室温,过滤除去不溶物,滤饼用乙醇(20毫升 x 2)清洗,滤液减压浓缩得到粗产品。粗产品用二氯甲烷(300毫升)溶解,过滤除去不溶物,滤饼用二氯甲烷(20毫升 x 2)清洗,滤液减压浓缩得到粗产品13-3(淡黄色油状液体,4.1克)。该化合物不需进一步纯化,直接用于下一步合成。MS(ESI,m/z):274.2[M+H]
+。
步骤4
在25摄氏度,氮气保护搅拌条件下,向50毫升三口烧瓶中加入化合物13-3(2.0克,7.0毫摩尔,1.0当量),N-甲基吡咯烷酮(20.0毫升),N,N-二异丙基乙胺(1.81克,14毫摩尔,2当量)和2-(2,6-二氧代-哌啶-3-基)-4-氟-异吲哚-1,3-二酮(2.2克,7.6毫摩尔,1.1当量)。所得混合物在90摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相色谱(C18)进行纯化,在30分钟内,用10%→95%的甲醇/水(0.1%三氟乙酸)流动相进行洗脱,检测器UV 254/220纳米;得到化合物13-4(黄色固体,1.4克,产率38%)。MS(ESI,m/z):530.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.94(s,1H),7.54–7.48(m,1H),7.14–7.12(m,1H),6.92–6.89(m,1H),6.54–6.51(m,1H),4.97–4.84(m,1H),3.72–3.68(m,4H),3.55–3.41(m,6H),2.94–2.61(m,5H),2.53–2.49(m,4H),2.22–2.12(m,1H),1.48(s,9H)。
步骤5
在25摄氏度搅拌条件下,向化合物13-4(500.0毫克,0.9毫摩尔,1.0当量)的二氯甲烷(8.0毫升)溶液中加入三氟乙酸(2.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品13-5(黄色固体,740.0毫克)。该化 合物不需进一步纯化,直接用于后续合成。MS(ESI,m/z):430.2[M+H]
+。
步骤6
在25摄氏度搅拌条件下,向化合物2-(烯丙基氧基)乙烷-1-醇(8.0克,77.5毫摩尔,1.0当量)和吡啶(18.6克,232.6毫摩尔,3.0当量)的二氯甲烷(50.0毫升)溶液中分批加入对甲苯磺酰氯(22.4克,116.3毫摩尔,1.5当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→30%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物13-6(无色液体,20.0克,产率99%)。MS(ESI,m/z):257.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.86–7.80(m,2H),7.39–7.33(m,2H),5.91–5.78(m,1H),5.35–5.15(m,2H),4.24–4.16(m,2H),3.98–3.95(m,2H),3.69–3.62(m,2H),2.47(s,3H)。
步骤7
在25摄氏度搅拌条件下,向化合物13-6(19.0克,73.4毫摩尔,1.0当量)的乙腈(100.0毫升)溶液中加入碳酸钾(30.7克,220.15毫摩尔,3.0当量)和3-(甲胺基)-1-丙醇(85.9克,95.4毫摩尔,1.3当量)。所得混合物在85摄氏度下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→30%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物13-7(淡黄色油状液体,11.0克,产率86%)。MS(ESI,m/z):174.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ5.99–5.87(m,1H),5.34–5.17(m,2H),4.02–3.99(m,2H),3.85–3.77(m,2H),3.58(t,J=5.8Hz,2H),2.70–2.64(m,4H),2.34(s,3H),1.78–1.67(m,2H).
步骤8
在25摄氏度,氮气保护搅拌条件下,向化合物3-2(1.5克,2.8毫摩尔,1.0当量),氟化铯(0.9克,5.6毫摩尔,2.0当量)和N,N-二异丙基乙胺(1.6毫升,8.4毫摩尔,3.0当量)的N-甲基吡咯烷 酮(15.0毫升)溶液中加入化合物13-7(1.0克,5.6毫摩尔,2.0当量)。所得混合物在130摄氏度下搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至室温。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用50%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物13-8(黄色油状液体,1.1克,产率61%)。MS(ESI,m/z):642.2/644.2/646.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.70(d,J=2.0Hz,1H),5.98–5.85(m,1H),5.33–5.11(m,2H),4.48(t,J=6.6Hz,2H),4.33–4.29(m,4H),4.01–3.99(m,2H),3.57–3.53(m,4H),2.65–2.61(m,4H),2.32(s,3H),2.09–1.94(m,4H),1.85–1.81(m,2H),1.53(s,9H).
步骤9
在25摄氏度,氮气保护搅拌条件下,向化合物13-8(1.2克,1.8毫摩尔,1.0当量)的四氢呋喃/水(10/1,13.2毫升)溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(0.7克,2.7毫摩尔,1.5当量),磷酸钾(0.8克,3.5毫摩尔,2.0当量)和氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(139.3毫克,0.2毫摩尔,0.1当量)。所得混合物在60摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物13-9(淡黄色固体,710.0毫克,产率57%)。MS(ESI,m/z):706.4/708.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.78–7.71(m,2H),7.44–7.39(m,1H),7.34–7.31(m,2H),7.24–7.21(m,1H),7.18–7.15(m,1H),5.92–5.79(m,1H),5.26–5.10(m,2H),4.53–4.33(m,6H),3.96–3.93(m,2H),3.65–3.62(m,4H),2.83–2.81(m,4H),2.47(s,3H),2.23–2.09(m,2H),1.99–1.97(m,2H),1.85–1.83(m,2H),1.55(s,9H)。
步骤10
通过制备级手性高效液相色谱法对步骤9所得化合物13-9(710.0毫克)进行手性拆分:手性柱NB-Lux 5 m i-Cellulose-5,2.12 x 25厘米,5微米;流动相A:正己烷/二氯甲烷=5:1(0.5%2摩尔/升氨-甲醇),流动相B:乙醇;流速:20毫升/分钟;在9分钟内用20%的B相进行洗脱,检测器UV 220/254纳米。得到两个产品,较短保留时间(5分钟)的产品为化合物13-9a(淡黄色固体,310.0毫克,回收率44%),化合物13-9a:MS(ESI,m/z):706.4/708.4[M+H]
+;较长保留时间(7.2分 钟)的产品为化合物13-9b(淡黄色固体,300.0毫克,回收率42%),化合物13-9b:MS(ESI,m/z):706.4/708.4[M+H]
+。
步骤11
在零摄氏度搅拌条件下,向化合物13-9a(300.0毫克,0.4毫摩尔,1.0当量)的丙酮/水(4/1,3.8毫升)溶液中加入N-甲基吗啉氧化物(70.9毫克,0.6毫摩尔,1.5当量)和锇酸钾二水合物(14.9毫克,0.04毫摩尔,0.1当量)。所得混合物在零摄氏度下搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用50%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物13-10a(棕色固体,160.0毫克,产率54%)。MS(ESI,m/z):740.4/742.4[M+H]
+。
步骤12
在25摄氏度搅拌条件下,向化合物13-10a(60.0毫克,0.1毫摩尔,1.0当量)的乙腈/水(4/1,2.5毫升)溶液中加入高碘酸钠(104.0毫克,0.5毫摩尔,6.0当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,加入水(10.0毫升)稀释反应液,然后用二氯甲烷(10毫升 x 3)萃取混合液,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品13-11a(淡黄色油状液体,65.0毫克),粗品直接用于下一步反应。MS(ESI,m/z):708.4/710.4[M+H]
+。
步骤13
在25摄氏度搅拌条件下,向化合物13-5(47.3毫克,0.11毫摩尔,1.2当量),醋酸钠(7.5毫克,0.09毫摩尔,1.0当量),醋酸(0.01毫升,0.09毫摩尔,1.0当量)和氰基硼氢化钠(11.5毫克,0.18毫摩尔,2.0当量)的甲醇(2.0毫升)溶液中加入化合物13-11a(65.0毫克,0.09毫摩尔,1.0当量)。 所得混合物在25摄氏度下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物13-12a(黄色固体,40毫克,产率42%)。MS(ESI,m/z):1121.4/1123.4[M+H]
+。
步骤14
在25摄氏度搅拌条件下,向化合物13-12a(40.0毫克,0.03毫摩尔,1.0当量)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(0.5毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→33%的B相梯度洗脱;检测器UV 254/220纳米;得到化合物13a(黄色固体,32.0毫克,产率59%)。MS(ESI,m/z):1021.4/1023.4[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.01(d,J=1.6Hz,1H),7.76(d,J=8.3Hz,1H),7.58–7.54(m,1H),7.46–7.40(m,1H),7.31–7.17(m,3H),7.13–7.01(m,3H),5.12–4.99(m,1H),4.72(d,J=14.3Hz,2H),4.63–4.57(m,2H),4.27(s,2H),3.99–3.82(m,4H),3.81–3.61(m,6H),3.51–3.37(m,10H),3.28–3.05(m,8H),2.99(s,3H),2.88–2.63(m,3H),2.38–2.29(m,2H),2.21–2.04(m,5H);
19F NMR(282MHz,CD
3OD)δ-77.08,-122.94,-122.97。
步骤11’
在零摄氏度搅拌条件下,向化合物13-9b(300.0毫克,0.4毫摩尔,1.0当量)的丙酮/水(4/1,5.0毫升)溶液中加入N-甲基吗啉氧化物(70.9毫克,0.6毫摩尔,1.5当量)和锇酸钾二水合物(14.9毫克,0.04毫摩尔,0.1当量)。所得混合物在零摄氏度下搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用50→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器,UV254纳米;得到化合物13-10b(棕色固体,136.0毫克,产率45%)。MS(ESI,m/z):740.4/742.4[M+H]
+。
步骤12’
在25摄氏度搅拌条件下,向化合物13-10b(60.0毫克,0.1毫摩尔,1.0当量)的乙腈/水(4/1,2.5毫升)溶液中加入高碘酸钠(104.0毫克,0.5毫摩尔,6.0当量)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,加入水(10.0毫升)稀释反应液,然后用二氯甲烷(10毫升 x 3)萃取混合液,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品13-11b(淡黄色油状液体,62.0毫克),粗品直接用于下一步反应。MS(ESI,m/z):708.2/710.2[M+H]
+。
步骤13’
在25摄氏度搅拌条件下,向化合物13-5(45.1毫克,0.10毫摩尔,1.2当量),醋酸钠(7.2毫克,0.08毫摩尔,1.0当量),醋酸(5.3毫克,0.08毫摩尔,1.0当量)和氰基硼氢化钠(11.0毫克,0.17毫摩尔,2.0当量)的甲醇(2.0毫升)溶液中加入化合物13-11b(62.0毫克,0.08毫摩尔,1.0当量)。所得混合物在25摄氏度下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物13-12b(黄色固体,45.0毫克,产率45%)。MS(ESI,m/z):1121.4/1123.4[M+H]
+。
步骤14’
在25摄氏度搅拌条件下,向化合物13-12b(45.0毫克,0.04毫摩尔,1.0当量)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(0.5毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→33%B相梯度洗脱;检测器UV 254/220纳米;得到化合物13b(黄色固体,33.0毫克,产率54%)。MS(ESI,m/z):1021.4/1023.4 [M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.08(d,J=1.7Hz,1H),7.83(d,J=8.3Hz,1H),7.65–7.60(m,1H),7.52–7.47(m,1H),7.37–7.23(m,3H),7.17–7.09(m,3H),5.18–5.07(m,1H),4.79(d,J=14.2Hz,2H),4.67(t,J=5.8Hz,2H),4.34(s,2H),4.04–3.89(m,4H),3.87–3.82(m,4H),3.78–3.73(m,2H),3.65–3.50(m,6H),3.45(s,3H),3.36–3.22(m,6H),3.16(s,2H),3.06(s,3H),2.98–2.67(m,4H),2.44–2.33(m,2H),2.24(s,4H),2.20–2.09(m,1H);
19F NMR(282MHz,CD
3OD)δ-77.09,-122.95,-122.98。
实施例14
4-((5-(1-(5-((3-((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)戊基)哌啶-4-基)戊基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮三盐酸盐14。
步骤1
在25摄氏度搅拌条件下,向化合物4-(3-羟丙基)哌啶-1-羧酸叔丁酯(15.0克,58.6毫摩尔,1.0当量)的二氯甲烷(300.0毫升)溶液中加入氯铬酸吡啶盐(25.8克,117.1毫摩尔,2.0当量)。所得混合物在25摄氏度下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,过滤除去不溶物,滤饼用二氯甲烷(2 x 20毫升)清洗,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→50%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物14-1(淡黄色油,6.0克,产率38%)。MS(ESI,m/z):242.3[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ9.68(t,J=1.6Hz,1H),3.92(d,J=13.1Hz,2H),2.65(s,2H),2.47–2.43(m,2H),1.66–1.56(m,2H),1.51–1.43(m,2H),1.39(s,9H),1.35–1.30(m,1H),1.02–0.86(m,2H).
步骤2
在-78摄氏度,氮气保护搅拌条件下,向化合物氰甲基磷酸二乙酯(4.4克,23.6毫摩尔,1.2当量)的四氢呋喃(6毫升)溶液中依次滴加双三甲基硅基胺基锂(1摩尔/升的四氢呋喃溶液,25.2毫 升,25.2毫摩尔,1.2当量)和化合物14-1(5.5克,20.5毫摩尔,1.0当量)的四氢呋喃溶液(2.0毫升)。所得混合物在-78摄氏度下搅拌反应40分钟,反应过程通过液质和薄层层析来监控。反应完全后,在零摄氏度下,向反应液中加入饱和氯化铵溶液(30毫升)淬灭反应,然后用二氯甲烷(50毫升 x 3)萃取反应体系,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→30%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物14-2(淡黄色液体,3.8克,产率67%)。MS(ESI,m/z):265.2[M+H]
+。
步骤3
在25摄氏度,氮气保护搅拌条件下,向化合物14-2(3.8克,13.7毫摩尔,1.0当量)的甲醇(35.0毫升)溶液中加入雷尼镍(380.0毫克)和氨的甲醇溶液(7摩尔/升,10毫升)。所得混合物在1个大气压的氢气氛围下,于25摄氏度搅拌反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,用硅藻土过滤除去镍催化剂,滤液减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物14-3(黄色液体,2.2克,产率57%)。MS(ESI,m/z):271.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.08(d,J=13.0Hz,2H),2.76–2.60(m,4H),1.65(d,J=13.3Hz,2H),1.51–1.42(m,12H),1.37–1.30(m,6H),1.29–1.21(m,2H),1.15–1.02(m,2H)。
步骤4
在25摄氏度,氮气保护搅拌条件下,向50毫升三口烧瓶中加入化合物14-3(1.1克,3.9毫摩尔,1.1当量),N-甲基吡咯烷酮(8.0毫升),N,N-二异丙基乙胺(928.8毫克,7.2毫摩尔,2.0当量)和2-(2,6-二氧代-哌啶-3-基)-4-氟-异吲哚-1,3-二酮(1.0克,3.6毫摩尔,1.0当量)。混合物在90摄氏度下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相快速色谱(C18)进行纯化,在30分钟内,用10%→95%的乙腈/水(0.1%三氟乙酸)流动相进行洗脱,检测器UV 254/220纳米;得到化合物14-4(黄色固体,647.0毫克,产率34%)。MS(ESI,m/z):527.2[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向化合物14-4(300.0毫克,0.57毫摩尔,1.0当量)的二氯甲烷(9.0毫升)溶液中加入三氟乙酸(3.0毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品14-5(黄色固体,364.0毫克)。该粗产品直接用于下一步合成。MS(ESI,m/z):427.2[M+H]
+。
步骤6
在25摄氏度搅拌条件下,向化合物14-5(30.9毫克,0.07毫摩尔,1.2当量),醋酸钠(7.4毫克,0.09毫摩尔,1.5当量),醋酸(5.2毫克,0.09毫摩尔,1.5当量)和氰基硼氢化钠(11.4毫克,0.17毫摩尔,3.0当量)的甲醇(2.0毫升)溶液中滴加化合物3-6a(45.0毫克,0.06毫摩尔,1.0当量)的甲醇(0.5毫升)溶液。所得混合物在50摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液冷却至室温,反应液减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物14-6a(黄色固体,40毫克,产率59%)。MS(ESI,m/z):1116.6/1118.6[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物14-6a(40.0毫克,0.036毫摩尔,1.0当量)的甲醇(1.5毫升)溶液中加入盐酸的1,4-二氧六环溶液(4摩尔/升,1.5毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.1%盐酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→35%B相梯度洗脱;检测器:UV 254/220纳米;得到化合物14(黄色固体,8.0毫克,产率19%)。MS(ESI,m/z):1016.4/1018.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6,重水交换)δ7.99(s,1H),7.82(d,J=8.3Hz,1H),7.59(m,1H),7.46(m,1H),7.31(d,J=2.4Hz,1H),7.27–7.17(m,2H),7.12–7.01(m, 3H),5.07–4.98(m,1H),4.58–4.39(m,4H),4.19(s,2H),3.87(t,J=13.0Hz,2H),3.40(s,1H),3.33–3.27(m,3H),3.23–3.01(m,3H),2.99–2.93(m,2H),2.89–2.80(m,2H),2.78(s,3H),2.62(s,1H),2.24–2.18(m,2H),2.11–1.88(m,6H),1.86–1.76(m,2H),1.73–1.65(m,4H),1.60–1.56(m,3H),1.48–1.15(m,12H);
19F NMR(377MHz,DMSO-d
6)δ-122.04。
实施例15
4-((4-(4-(4-((3-((4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-8-基)-6-氯-8-氟-7-((S或R)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮四三氟乙酸盐15a;4-((4-(4-(4-((3-((4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-8-基)-6-氯-8-氟-7-((R或S)-3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮四三氟乙酸盐15b。
步骤1
在零摄氏度,氮气保护搅拌条件下,向化合物1-6(1.50克,4.313毫摩尔,1.00当量)和三乙胺(1.31克,12.940毫摩尔,3.00当量)的1,4-二氧六环(15.0毫升)溶液中滴加3,8-二氮杂双环[3.2.1]辛烷-3-甲酸叔丁酯(0.92克,4.313毫摩尔,1.00当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→28%乙酸乙酯/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物15-1(类白色固体,2.2克,产率91%)。MS(ESI,m/z):505.0/507.0/509.0[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.78(d,J=2.0Hz,1H),4.99–4.84(m,2H),4.18–3.90(m,2H),3.47–3.18(m,2H),2.13–1.82(m,4H),1.49(s,9H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向化合物15-1(2.10克,3.941毫摩尔,1.00当量)的N-甲基吡咯烷酮(42毫升)溶液中加入氟化铯(1.26克,7.882毫摩尔,2.0当量),4-1(0.98克,5.912毫摩尔,1.5当量)和N,N-二异丙基乙胺(1.61克,11.823毫摩尔,3.0当量)。所得混合物在130摄氏度下搅拌反应6小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的甲醇/水流动相(0.1%碳酸氢铵)进行洗脱;检测器,UV254纳米;得到化合物15-2(无色油,960毫克,产率38%)。MS(ESI,m/z):626.2/628.2/630.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.73(d,J=2.3Hz,1H),5.86–5.73(m,1H),5.06–4.91(m,2H),4.90–4.75(m,2H),4.54–4.43(m,2H),4.08–3.86(m,2H),3.47–3.20(m,2H),2.73–2.57(m,2H),2.49–2.39(m,2H),2.31(s,3H),2.10–2.02(m,4H),2.01–1.93(m,2H),1.90–1.80(m,2H),1.66–1.56(m,2H),1.49(s,9H)。
步骤3
在25摄氏度,氮气保护搅拌条件下,向化合物15-2(860.00毫克,1.372毫摩尔,1.00当量)的四氢呋喃/水(10/1,8.8毫升)的混合溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(555.79毫克,2.058毫摩尔,1.50当量),磷酸钾(582.29毫克,2.744毫摩尔,2.00当量)和氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(107.78毫克,0.137毫摩尔,0.10当量)。反应液在130摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物15-3(类白色固体,840毫克,产率88%)。MS(ESI,m/z):690.4/692.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.81(d,J=1.6Hz,1H),7.76(d,J=8.1Hz,1H),7.47–7.40(m,1H),7.34–7.31(m,1H),7.30–7.28(m,1H),7.27–7.19(m,1H),7.14(d,J=2.4Hz,1H),5.85–5.68(m,1H),5.04–4.88(m,4H),4.55–4.45(m,2H),4.15–3.91(m,2H),3.50–3.28(m,2H),2.78–2.64(m,2H),2.58–2.44(m,2H),2.35(s,3H),2.16–2.00(m,6H),1.97–1.86(m,2H),1.69–1.59(m,2H),1.56–1.50(s,9H)。
步骤4
通过制备级手性高效液相色谱法对步骤3所得化合物15-3(840毫克)进行手性拆分:手性柱NB-Lux 5 m i-Cellulose-5,2.12 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/升氨水),流动相B:异丙醇;流速:18毫升/分钟;在17分钟内用50%的B相进行洗脱,检测器UV 230/210纳米。得到两个产品,较短保留时间(6.0分钟)的产品为化合物15-3a(粉色固体,235毫克,回收率29%),化合物15-3a:MS(ESI,m/z):690.4/692.3[M+H]
+;较长保留时间(13.2分钟)的产品为化合物15-3b(粉色固体,300毫克,回收率37%),化合物15-3b:MS(ESI,m/z):690.4/692.3[M+H]
+。
步骤5
在0摄氏度搅拌条件下,向化合物15-3a(235.00毫克,0.340毫摩尔,1.00当量)的水/四氢呋喃(1/4,5毫升)的溶液中加入N-甲基吗啉氧化物(59.82毫克,0.511毫摩尔,1.50当量)和锇酸钾二水合物(50.18毫克,0.136毫摩尔,0.40当量)。混合物在0摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物15-4a(粉色固体,170毫克,产率68%)。
MS(ESI,m/z):724.4/726.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.79(d,J=1.7Hz,1H),7.73(d,J=8.3Hz,1H),7.45–7.36(m,1H),7.33–7.27(m,2H),7.24–7.17(m,1H),7.15–7.10(m,1H),5.00–4.84(m,2H),4.58–4.40(m,2H),4.10–4.01(m,1H),3.98–3.89(m,1H),3.64–3.48(m,2H),3.43–3.23(m,3H),2.78–2.53(m,2H),2.50–2.38(m,2H),2.31–2.23(m,3H),2.12–1.81(m,7H),1.73–1.57(m,3H),1.50(s,9H)。
步骤6
在零摄氏度搅拌条件下,向化合物15-4a(70.00毫克,0.092毫摩尔,1.00当量)的水/四氢呋喃 (1/4,5毫升)的混合溶液中加入高碘酸钠(157.81毫克,0.580毫摩尔,6.00当量)。混合物在零摄氏度下反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,加入水(50毫升)稀释反应液,然后用二氯甲烷(50毫升 x 3)萃取。合并的有机相用无水硫酸钠干燥,过滤,滤液浓缩得化合物15-5a的粗产品(类白色固体,61毫克)。粗产品直接用于下一步反应,未进一步纯化。MS(ESI,m/z):692.3/694.3[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物15-5a(61毫克,0.084毫摩尔,1.00当量),醋酸(5.28毫克,0.088毫摩尔,1.05当量)和氰基硼氢化钠(11.10毫克,0.177毫摩尔,2.11当量)的甲醇(2毫升)溶液中滴加化合物1-4(30.97毫克,0.071毫摩尔,0.85当量)的甲醇(1毫升)溶液。所得混合物在25摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物15-6a(黄色固体,61毫克,产率62%)。MS(ESI,m/z):1089.6/1091.6.[M+H]
+。
步骤8
在25摄氏度搅拌条件下,向化合物15-6a(61.00毫克,0.053毫摩尔,1.00当量)的二氯甲烷(3毫升)溶液中加入三氟乙酸(1毫升)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:甲醇;流速:25毫升/分钟;梯度在7分钟内用25%→54%B相梯度洗脱;检测器:UV 254/220纳米;得到15a(淡黄色固体,25毫克,产率31%)。MS(ESI,m/z):989.6/991.5[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.06(d,J=1.6Hz,1H),7.76(d,J=8.3Hz,1H),7.58–7.52(m,1H),7.45–7.38(m,1H),7.28(d,J=2.4Hz,1H),7.25–7.16(m,2H),7.10–7.02(m,3H),5.22–5.14(m,2H),5.09–5.01(m,1H),4.66–4.55(m,2H),3.71(d,J=12.8Hz,2H),3.49–3.42(m,3H),3.37(t,J=6.4Hz,3H),3.30–3.20(m,7H),3.13–2.98(m,5H),2.92(s,3H),2.88–2.78(m,3H),2.77–2.66(m,2H),2.41–2.26(m,4H),2.21–2.05(m,3H),1.89–1.74(m,4H),1.74–1.63(m,4H);
19F NMR(377MHz,CD
3OD)δ-77.15,-123.17。
步骤5’
在零摄氏度搅拌条件下,向化合物15-3b(300.00毫克,0.435毫摩尔,1.00当量)的水/四氢呋喃(1/4,5毫升)的混合溶液中加入N-甲基吗啉氧化物(76.37毫克,0.652毫摩尔,1.5当量)和锇酸钾二水合物(64.05毫克,0.174毫摩尔,0.4当量)。混合物在零摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物15-4b(粉色固体,220毫克,产率69%)。MS(ESI,m/z):724.4/726.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.82(d,J=1.7Hz,1H),7.77(d,J=8.3Hz,1H),7.44(t,J=7.6Hz,1H),7.33(d,J=7.7Hz,2H),7.28–7.21(m,1H),7.17–7.13(m,1H),5.06–4.84(m,2H),4.60–4.40(m,2H),4.21–4.04(m,1H),4.02–3.88(m,1H),3.68–3.50(m,2H),3.48–3.25(m,3H),2.80–2.54(m,2H),2.53–2.40(m,2H),2.34–2.26(m,3H),2.16–1.85(m,7H),1.76–1.63(m,3H).1.52(s,9H)。
步骤6’
在零摄氏度搅拌条件下,向化合物15-4b(70.00毫克,0.092毫摩尔,1.00当量)的水/四氢呋喃(1/4,5毫升)的混合溶液中加入高碘酸钠(117.83毫克,0.551毫摩尔,6.00当量)。混合物在25摄氏度下反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,加入水(50毫升)稀释反应液,然后用二氯甲烷(50毫升 x 3)萃取。合并的有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液浓缩得化合物15-5b的粗产品(黄色固体,69毫克)。粗产品未进一步纯化,直接用于下一步反应。MS(ESI,m/z):692.3/694.3[M+H]
+。
步骤7’
在25摄氏度搅拌条件下,向化合物15-5b(69.00毫克,0.095毫摩尔,1.00当量),醋酸(5.99毫克,0.100毫摩尔,1.0当量)和氰基硼氢化钠(12.53毫克,0.199毫摩尔,2.0当量)的甲醇(2毫 升)溶液中滴加化合物1-4(37.09毫克,0.090毫摩尔,0.90当量)的甲醇(1毫升)溶液。所得混合物在25摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物15-6b(黄色固体,45毫克,产率43%)。MS(ESI,m/z):1089.5/1091.5[M+H]
+。
步骤8’
在25摄氏度搅拌条件下,向化合物15-6b(45.00毫克,0.039毫摩尔,1.00当量)的二氯甲烷(3毫升)溶液中加入三氟乙酸(1毫升)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:甲醇;流速:25毫升/分钟;梯度在7分钟内用25%→54%的B相梯度洗脱;检测器UV254/220纳米;得到15b(淡黄色固体,14.8毫克,产率25%)。MS(ESI,m/z):[M+H]
+=989.6/991.5;
1H NMR(300MHz,CD
3OD)δ8.09(d,J=1.7Hz,1H),7.78(d,J=8.3Hz,1H),7.61–7.55(m,1H),7.48–7.40(m,1H),7.30(d,J=2.5Hz,1H),7.22(d,J=5.8Hz,2H),7.12–7.03(m,3H),5.27–5.16(m,2H),5.12–5.01(m,1H),4.70–4.57(m,2H),3.74(d,J=12.7Hz,2H),3.53–3.44(m,3H),3.44–3.37(m,3H),3.30–3.21(m,5H),3.16–2.98(m,6H),2.95(s,3H),2.91–2.67(m,6H),2.46–2.27(m,4H),2.22–2.07(m,3H),1.93–1.68(m,8H);
19F NMR(377MHz,CD
3OD)δ-77.20,-123.15。
实施例16
4-((3-(4-(3-((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丙基)哌嗪-1-基)丙基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮四盐酸盐16。
步骤1:
在25摄氏度氮气保护搅拌条件下,向1-溴-3-羟基萘(10克,44.38毫摩尔,1.0当量)的二氯甲烷(100毫升)溶液中加入N,N-二异丙基乙胺(8.69克,66.57毫摩尔,1.5当量)和氯甲基甲醚(4.69克,57.69毫摩尔,1.3当量)。混合物在25摄氏度下反应3小时,反应过程通过液质监控。反应结束后,将反应液浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→10%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物16-1(白色固体,10.5克,产率87%)。
1H NMR(300MHz,CDCl
3)δ8.16–8.11(m,1H),7.75–7.71(m,1H),7.57(d,J=2.4Hz,1H),7.50–7.43(m,2H),7.39(d,J=2.4Hz,1H),5.28(s,2H),3.52(s,3H)。
步骤2:
在25摄氏度氮气保护搅拌条件下,向16-1(10克,35.56毫摩尔,1.0当量)的1,4-二氧六环(100毫升)溶液中依次加入醋酸钾(14.70克,142.26毫摩尔,4.0当量),联硼酸频那醇酯(12.36克,46.23毫摩尔,1.3当量)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(3.05克,3.55毫摩尔,0.1当量)。混合物在氮气氛围下100摄氏度反应1小时,反应过程通过液质监控。反应结束后,混合物冷却至25摄氏度,减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→20%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物16-2(白色固体,10克,产率85%)。MS(ESI,m/z):315.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.72–8.66(m,1H),7.82(d,J=2.7Hz,1H),7.79–7.73(m,1H),7.51(d,J=2.7Hz,1H),7.49–7.40(m,2H),5.33(s,2H),3.54(s,3H),1.44(s,12H)。
步骤3:
在25摄氏度氮气保护条件下,向250毫升三口瓶中依次加入化合物3-2(9克,16.89毫摩尔,1.0当量),16-2(5.59克,16.89毫摩尔,1.0当量),1,4-二氧六环(80毫升),水(20毫升),碳酸钠 (3.77克,33.78毫摩尔,2.0当量),三二亚苄基丙酮二钯(0)(775毫克,0.85毫摩尔,0.05当量),和4-(2,6-二甲氧基苯基)-3-叔丁基-2,3-二氢-1,3-苯并氧磷杂环(558毫克,1.69毫摩尔,0.1当量)。混合物在氮气氛围下60摄氏度反应2小时。反应过程通过液质监控。反应结束后,将反应液浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→20%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂得粗产品。粗产品再通过反相色谱柱(C18柱)进行纯化,在20分钟内用40%→90%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254/220纳米;得到化合物16-3(两个立体异构体的外消旋混合物,黄色固体,6克,产率55%)。MS(ESI,m/z):613.2/615.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.88–7.85(m,2H),7.58(d,J=2.5Hz,1H),7.53–7.47(m,1H),7.32–7.30(m,2H),7.21(d,J=2.4Hz,1H),5.36(s,2H),4.56–4.41(m,4H),3.77–3.65(m,2H),3.58(s,3H),2.06–2.00(m,2H),1.87–1.79(m,2H),1.56(s,9H)。
步骤4:
通过制备级超临界液相色谱法对步骤3所得化合物16-3(6克)进行手性拆分:手性柱NB_CHIRALPAK IC,5 x 25厘米,5微米;流动相A:超临界二氧化碳流体,流动相B:乙醇;流速:160毫升/分钟;柱温:35摄氏度;用45%流动相B洗脱15分钟;检测器UV225纳米。得到两个产品,较短保留时间(9.03分钟)的产品为化合物16-3a(黄色固体,2.5克,回收率41%),化合物16-3a:MS(ESI,m/z):613.2/615.2[M+H]
+;较长保留时间(10.78分钟)的产品为化合物为16-3b(黄色固体,2.9克,回收率48%),化合物16-3b:MS(ESI,m/z):613.2/615.2[M+H]
+。
步骤5
在零摄氏度,氮气保护搅拌条件下,向化合物16-3a(300.0毫克,0.47毫摩尔,1.0当量)和3-丁烯1-醇(42.3毫升,0.56毫摩尔,1.2当量)的四氢呋喃(5.0毫升)溶液中滴加1摩尔/升的叔丁醇钾的四氢呋喃溶液(0.59毫升,0.56毫摩尔,1.2当量)。所得混合物在零摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析 法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物16-4a(类白色固体,230.0毫克,产率72%)。MS(ESI,m/z):649.3/651.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.89–7.79(m,2H),7.56(d,J=2.4Hz,1H),7.52–7.45(m,1H),7.39–7.29(m,2H),7.22(d,J=2.5Hz,1H),6.02–5.88(m,1H),5.40–5.33(m,2H),5.25–5.08(m,2H),4.56–4.36(m,6H),3.59–3.57(m,5H),2.71–2.57(m,2H),2.07–2.01(m,2H),1.91–1.87(m,2H),1.55(s,9H)。
步骤6
在零摄氏度搅拌条件下,向化合物16-4a(225.0毫克,0.33毫摩尔,1.0当量)的丙酮/水(4/1,5.0毫升)溶液中加入N-甲基吗啉氧化物(60.9毫克,0.49毫摩尔,1.5当量)和锇酸钾二水合物(12.7毫克,0.33毫摩尔,0.1当量)。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用40%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物16-5a(类白色固体,180毫克,产率76%)。MS(ESI,m/z):683.3/685.3[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物16-5a(60.0毫克,0.08毫摩尔,1.0当量)的乙腈/水(4/1,2.5毫升)溶液中加入高碘酸钠(107.1毫克,0.5毫摩尔,6.0当量)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入水(10.0毫升),用二氯甲烷(10.0毫升 x 3)萃取混合液,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品16-6a(淡黄色油状液体,62.0毫克),粗品直接用于下一步反应。MS(ESI,m/z):651.3/653.3[M+H]
+。
步骤8
在25摄氏度搅拌条件下,向化合物9-6(53.8毫克,0.11毫摩尔,1.2当量),醋酸钠(15.6毫克,0.19毫摩尔,2.0当量),醋酸(17.2毫克,0.29毫摩尔,3.0当量)和氰基硼氢化钠(18.0毫克,0.29毫摩尔,3.0当量)的甲醇(2.0毫升)溶液中滴加化合物16-6a(62.0毫克,0.10毫摩尔,1.0当量)的甲醇(0.5毫升)溶液。所得混合物在25摄氏度下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物16-7a(黄色固体,37毫克,产率35%)。MS(ESI,m/z):1105.6/1107.6[M+H]
+。
步骤9
在25摄氏度搅拌条件下,向化合物16-7a(37.0毫克,0.03毫摩尔,1.0当量)的甲醇(2.0毫升)溶液中加入4摩尔/升的盐酸的1,4-二氧六环溶液(2.0毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.05%盐酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→30%B相梯度洗脱;检测器UV 254/220纳米;得到化合物16(黄色固体,24.0毫克,产率64%)。MS(ESI,m/z):961.4/963.4[M+H]
+;
1H NMR(300MHz,DMSO-d
6,重水交换)δ8.05(s,1H),7.89–7.86(d,J=8.3Hz,1H),7.70–7.66(m,1H),7.55–7.50(m,1H),7.38–7.37(m,1H),7.33–7.21(m,3H),7.14–7.12(m,2H),5.13–5.07(m,1H),4.65–4.47(m,4H),4.25(s,2H),3.96–3.88(m,4H),3.52–3.48(m,4H),3.39–3.24(m,7H),3.18–3.08(m,1H),2.99–2.77(m,6H),2.75–2.70(m,1H),2.39–2.32(m,1H),2.31–2.15(m,5H),2.14–1.93(m,8H);
19F NMR(282MHz,DMSO-d
6)δ-122.00。
实施例17
4-((15-(((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-12-甲基-3,6,9-三氧-12-氮杂十五烷基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮甲酸盐17a;4-((15-(((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-12-甲基-3,6,9-三氧-12-氮杂十五烷基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚-1,3-二酮甲酸盐17b。
步骤1
在零摄氏度搅拌条件下,向反应瓶中依次加入化合物2-(2,6-二氧代-哌啶-3-基)-4-氟-异吲哚-1,3-二酮(2.00克,7.241毫摩尔,1.00当量),1-氨基-3,6,9-三噁-11-十一醇(1.54克,7.969毫摩尔,1.10当量),N,N-二甲基甲酰胺(20毫升)和N,N-二异丙基乙胺(1.87克,14.481毫摩尔,2.00当量)。所得混合物在90摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温。混合物用硅胶柱层析法纯化,用0%→8%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物17-1(黄色油,701毫克,产率21%)。MS(ESI,m/z):450.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.16(s,1H),7.52–7.47(m,1H),7.11(d,J=7.1Hz,1H),6.93(d,J=8.5Hz,1H),4.95–4.89(m,1H),3.76–3.66(m,12H),3.64–3.59(m,2H),3.48(t,J=5.3Hz,2H),2.92–2.85(m,1H),2.82–2.71(m,2H),2.18–2.10(m,1H)。
步骤2
在25摄氏度搅拌条件下,向化合物17-1(170.00毫克,0.367毫摩尔,1.00当量)的二氯甲烷(150.0毫升)溶液中加入1,1,1-三乙酰氧基-1,1-二氢-1,2-苯碘氧基-3(1H)-酮(327.60毫克,毫摩尔,2.00当量)。所得混合物在25摄氏度下搅拌反应16小时。反应过程通过液质和薄层层析来监控。反应结束后,过滤除去不溶物,滤饼用二氯甲烷(3 x 20毫升)洗涤。滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物17-2(淡黄色油,140毫克,产率70%)。MS(ESI,m/z):448.3[M+H]
+。
步骤3
在25摄氏度条件下,依次向25毫升单口瓶中加入化合物2-9a(25毫克,0.042毫摩尔,1.0当 量),17-2(56.3毫克,0.126毫摩尔,3.0当量),甲醇(2.0毫升),醋酸(3.78毫克,0.063毫摩尔,1.5当量)和氰基硼氢化钠(3.95毫克,0.063毫摩尔,1.5当量)。所得混合物在40摄氏度反应2小时。反应过程通过液质和薄层层析来监控。反应结束后减压浓缩除去多余的溶剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%三氟乙酸)进行洗脱;检测器UV254纳米;得到化合物17-3a(黄色油状液体,23毫克,产率48%)。MS(ESI,m/z):1027.4/1029.4[M+H]
+。
步骤4
在0摄氏度条件下,依次向25毫升单口瓶中加入化合物17-3a(20毫克,0.018毫摩尔,1.0当量),二氯甲烷(2.0毫升)和三氟乙酸(1.0毫升)。所得混合物在25摄氏度反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩除去多余的溶剂得到粗产品。粗产品通过制备级高效液相色谱仪进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米,流动相A:水(0.05%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→31%B相梯度洗脱;检测器UV 254/220纳米;得到化合物17a(黄色固体,10毫克,产率56%)。MS(ESI,m/z):927.4/929.4;
1H NMR(300MHz,DMSO-d
6)δ11.11(s,1H),10.30–9.80(m,1H),8.18(s,1H),7.94(d,J=1.6Hz,1H),7.81(d,J=8.3Hz,1H),7.59–7.52(m,1H),7.48–7.40(m,1H),7.28(d,J=2.4Hz,1H),7.25–7.17(m,2H),7.13–6.99(m,3H),6.62–6.53(m,1H),5.11–4.99(m,1H),4.35(t,J=6.6Hz,2H),3.85–3.77(m,4H),3.59–3.56(m,2H),3.55–3.46(m,9H),3.44–3.25(m,6H),3.03–2.96(m,4H),2.93–2.78(m,2H),2.63–2.53(m,2H),2.19(s,3H),2.08–1.96(m,1H),1.93–1.79(m,2H)。
步骤3’
在25摄氏度氮气保护条件下,依次向50毫升三口瓶中加入化合物2-8b(760毫克,1.1毫摩尔,1.0当量),1,3-二甲基巴比妥酸(279.8毫克,1.7毫摩尔,1.5当量),四(三苯基膦)钯(69.0毫克,0.057毫摩尔,0.05当量)和二氯甲烷(10毫升),所得混合物在25摄氏度反应5小时,反应过程通过液质和薄色谱层层析监控。反应结束后减压浓缩除去多余的溶剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%甲酸)进行洗脱,检测器UV254纳米,得到化合物2-9b(淡黄色固体,640毫克,产率89%)。MS(ESI,m/z):596.2/598.2 [M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.61(s,1H),7.69–7.66(m,2H),7.35–7.31(m,1H),7.28–7.27(m,1H),7.18–7.08(m,3H),4.52–4.49(m,2H),3.82–3.64(m,8H),3.26(brs,2H),3.00–2.91(m,2H),2.50(s,3H),2.21–2.18(m,2H),1.49(s,9H)。
步骤4’
在25摄氏度条件下,依次向25毫升单口瓶中加入化合物2-9b(25毫克,0.04毫摩尔,1.0当量),17-2(51.8毫克,0.12毫摩尔,3.0当量),甲醇(2.0毫升),醋酸(3.78毫克,0.06毫摩尔,1.5当量)和氰基硼氢化钠(3.95毫克,0.063毫摩尔,1.5当量)。所得混合物在40摄氏度反应2小时。反应过程通过液质和薄层层析来监控。反应结束后减压浓缩除去多余的溶剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%三氟乙酸)进行洗脱;检测器UV254纳米;得到化合物17-3b(黄色油状液体,25毫克,产率54%)。MS(ESI,m/z):1027.4/1029.4[M+H]
+。
步骤5’
在0摄氏度空气条件下,依次向25毫升单口瓶中加入化合物17-3b(20毫克,0.018毫摩尔,1.0当量),二氯甲烷(2.0毫升)和三氟乙酸(1.0毫升)。所得混合物在25摄氏度反应2小时。反应过程通过液质监控。反应结束后减压浓缩除去多余的溶剂得到粗产品。粗产品通过制备级高效液相色谱仪进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米,流动相A:水(0.05%甲酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→31%B相梯度洗脱;检测器UV 254/220纳米;得到化合物17b(黄色固体,10毫克,产率57%)。MS(ESI,m/z):927.4/929.4;
1H NMR(300MHz,DMSO-d
6)δ11.19(s,1H),10.30–9.80(m,1H),8.26(s,1H),8.02(d,J=1.6Hz,1H),7.88(d,J=8.3Hz,1H),7.67–7.58(m,1H),7.56–7.47(m,1H),7.36(d,J=2.4Hz,1H),7.34–7.24(m,2H),7.20–7.06(m,3H),6.71–6.58(m,1H),5.18–5.06(m,1H),4.50–4.37(m,2H),3.92–3.84(m,4H),3.68–3.63(m,3H),3.61–3.54(m,5H),3.52–3.28(m,9H),3.16–3.04(m,4H),2.98–2.84(m,2H),2.69–2.60(m,2H),2.27(s,3H),2.15–2.02(m,1H),2.00–1.88(m,2H)。
实施例18
(2S,4R)-1-((S)-2-(叔丁基)-18-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-15-甲基-4-氧代-6,9,12-三氧基-3,15-二氮杂十八烷基)-4-羟基-N-((S)-1-(4-(4- 甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺18a;(2S,4R)-1-((S)-2-(叔丁基)-18-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-15-甲基-4-氧代-6,9,12-三氧基-3,15-二氮杂十八烷基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺18b。
步骤1
在零摄氏度,氮气保护搅拌条件下,向化合物三甘醇(20克,126.52毫摩尔,1.00当量)的二氯甲烷(400毫升)溶液中滴加三氟化硼的乙醚溶液(0.40毫升,2.824毫摩尔,0.05当量)和重氮乙酸乙酯(7.6克,63.26毫摩尔,0.50当量)。在氮气保护搅拌条件下,混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用33%的二氯甲烷/乙酸乙酯流动相洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物18-1(淡黄色油,4克,产率14%)。MS(ESI,m/z):237.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.14(q,J=7.1Hz,2H),4.07(s,2H),3.71–3.61(m,6H),3.60(s,4H),3.55–3.49(m,2H),2.88(s,1H),1.21(t,J=7.1Hz,3H)。
步骤2
在零摄氏度,氮气保护搅拌条件下,向化合物18-1(4.00克,16.94毫摩尔,1.00当量)和N,N-二异丙基乙胺(3.00克,22.02毫摩尔,1.3当量)的二氯甲烷(20毫升)溶液中滴加甲基磺酰氯(2.44克,20.33毫摩尔,1.2当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物18-2(棕色油,3.6克,产率67%)。MS(ESI,m/z):315.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.37–4.32(m,2H),4.17(q,J=7.1Hz,2H),4.09(s,2H),3.76–3.71(m,2H),3.71–3.57(m,8H),3.04(s,3H),1.25(t,J=7.1Hz,3H)。
步骤3
在25摄氏度,氮气保护搅拌条件下,向化合物18-2(1.80克,5.726毫摩尔,1.00当量)和3-(甲胺基)-1-丙醇(641.19毫克,6.834毫摩尔,1.30当量)的乙腈(18毫升)溶液中加入碳酸钾(1.53克,10.513毫摩尔,2.00当量)。在氮气保护搅拌条件下,混合物在90摄氏度下反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→45%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV200纳米;得到化合物18-3(无色油,800毫克,产率39%)。MS(ESI,m/z):308.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.23(q,J=7.1Hz,2H),4.16(s,2H),3.83–3.58(m,12H),2.69–2.59(m,4H),2.31(s,3H),1.77–1.64(m,2H),1.30(t,J=7.1Hz,3H)。
步骤4
在25摄氏度,氮气保护搅拌条件下,向化合物1-7(800毫克,1.65毫摩尔,1.00当量)的N-甲基吡咯烷酮(10毫升)溶液中加入氟化铯(395.38毫克,2.47毫摩尔,1.50当量),18-3(582.43毫克,1.65毫摩尔,1.0当量)和N,N-二异丙基乙胺(336.40毫克,2.47毫摩尔,1.50当量)。所得混合物在100摄氏度下搅拌反应3小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用20%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物18-4(棕色油,250毫克,产率19%)。MS(ESI,m/z):750.2/752.1/754.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.72(d,J=2.0Hz,1H),4.52(t,J=6.5Hz,2H),4.23(q,J=7.2Hz,2H),4.16(s,2H),3.83–3.59(m,18H),2.86–2.57(m,4H),2.51–2.27(m,3H),2.03(s,2H),1.52(s,9H),1.30(t,J=7.1Hz,3H)。
步骤5
在25摄氏度搅拌条件下,向化合物18-4(250毫克,0.28毫摩尔,1.00当量)的水/甲醇/四氢呋喃(1/1/1,1.5毫升)的混合溶液中加入氢氧化锂(34.65毫克,1.38毫摩尔,5.0当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱; 检测器,UV254纳米;得到化合物18-5(无色油,210毫克,产率89%)。MS(ESI,m/z):722.1/724.2/726.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.69(d,J=2.0Hz,1H),4.57–4.49(m,2H),4.01(s,2H),3.86–3.73(m,6H),3.73–3.57(m,12H),3.11–2.96(m,4H),2.64(s,3H),2.29–2.15(m,2H),1.49(s,9H)。
步骤6
在25摄氏度搅拌条件下,向化合物18-5(210毫克,0.27毫摩尔,1.00当量)和2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(131.41毫克,0.33毫摩尔,1.2当量)的N,N-二甲基甲酰胺(2.0毫升)溶液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(138.54毫克,0.27毫摩尔,1.0当量)和N,N-二异丙基乙胺(148.89毫克,1.09毫摩尔,4.0当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相快速色谱柱(C18柱)进行纯化,在11分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物18-6(类白色固体,260毫克,产率77%)。MS(ESI,m/z):1148.4/1150.3/1152.3[M+H]
+。
步骤7
在25摄氏度,氮气保护搅拌条件下,向化合物18-6(230毫克,0.18毫摩尔,1.00当量)的1,4-二氧六环/水(1/1,6毫升)的溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(61.44毫克,0.216毫摩尔,1.2当量),碳酸钠(57.25毫克,0.51毫摩尔,2.85当量)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(II)(15.44毫克,0.02毫摩尔,0.10当量)。反应液在90摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度。反应液浓缩后通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物18-7(类白色固体,140毫克,产率57%)。MS(ESI,m/z):1212.4/1214.4[M+H]
+。
步骤8
通过制备级手性高效液相色谱法对步骤7所得化合物18-7(140毫克)进行手性拆分:手性柱CHIRALPAK IC,2 x 25厘米,5微米;流动相A:正己烷/二氯甲烷=3/1(0.1%二乙基胺),流动相B:乙醇;流速:15毫升/分钟;在26分钟内用50%的B相进行洗脱,检测器UV220/235纳米。得到两个产品,较短保留时间(8.11分钟)的产品为化合物18-7a(类白色固体,52毫克,回收率37%),化合物18-7a:MS(ESI,m/z):1212.4/1214.4[M+H]
+;较长保留时间(18分钟)的产品为化合物18-7b(类白色固体,57毫克,回收率40%),化合物18-7b:MS(ESI,m/z):1212.4/1214.4[M+H]
+。
步骤9
在25摄氏度搅拌条件下,向化合物18-7a(52.00毫克,0.043毫摩尔,1.00当量)的二氯甲烷(1.5毫升)溶液中加入三氟乙酸(0.5毫升)。混合物在25摄氏度下反应0.5小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;梯度:在2分钟内用10%的B相洗脱,后在2.5分钟内用10%→36%的B相梯度洗脱,最后在10.5分钟内用36%→65%的B相梯度洗脱;检测器UV 220纳米;得到18a(类白色固体,24.3毫克,产率49%)。MS(ESI,m/z):1112.6/1114.6.[M+H]
+;
1H NMR(300MHz,DMSO-d
6):δ10.02(s,1H),8.99(s,1H),8.44(d,J=7.6Hz,1H),7.92(d,J=1.6Hz,1H),7.81(d,J=8.3Hz,1H),7.50–7.32(m,6H),7.29(d,J=2.4Hz,1H),7.22(d,J=4.0Hz,2H),7.07(d,J=2.4Hz,1H),5.14(s,1H),4.90(t,J=7.1Hz,1H),4.54(d,J=9.5Hz,1H),4.44(t,J=8.2Hz,1H),4.36(t,J=6.5Hz,2H),4.28(s,1H),3.94(s,2H),3.78(d,J=5.7Hz,4H),3.63–3.41(m,13H),2.90(d,J=5.0Hz,4H),2.53–2.35(m,7H),2.45(s,3H),2.19(s,1H),2.12–1.97(m,3H),1.37(d,J=7.0Hz,3H),0.93(s,9H)。
步骤10
在25摄氏度搅拌条件下,向化合物18-7b((57.00毫克,0.047毫摩尔,1.00当量)的二氯甲烷(1.5毫升)溶液中加入三氟乙酸(0.5毫升)。混合物在25摄氏度下反应0.5小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;梯度:在2分钟内用10%B相洗脱,后在2.5分钟内用10%→36%的B相梯度洗脱,最后在10.5分钟内用36%→65%的B相梯度洗脱;检测器UV 220纳米;得到18b(类白色固体,24.3毫克,回收率45%)。MS(ESI,m/z):1112.6/1114.6;
1H NMR(300MHz,DMSO-d
6):δ10.02(s,1H),8.99(s,1H),8.44(d,J=7.6Hz,1H),7.92(d,J=1.6Hz,1H),7.81(d,J=8.3Hz,1H),7.50–7.32(m,6H),7.29(d,J=2.4Hz,1H),7.22(d,J=4.0Hz,2H),7.07(d,J=2.4Hz,1H),5.14(s,1H),4.90(t,J=7.1Hz,1H),4.54(d,J=9.5Hz,1H),4.44(t,J=8.2Hz,1H),4.36(t,J=6.5Hz,2H),4.28(s,1H),3.94(s,2H),3.78(d,J=5.7Hz,4H),3.63–3.41(m,13H),2.90(d,J=5.0Hz,4H),2.53–2.35(m,7H),2.45(s,3H),2.19(s,1H),2.12–1.97(m,3H),1.37(d,J=7.0Hz,3H),0.93(s,9H)。
实施例19
(2S,4R)-1-((S)-2-(叔丁基)-21-((6-氯-8-氟-7-((S或R)-3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-18-甲基-4-氧代-6,9,12,15-四氧代-3,18-二氮杂苯三酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺19a;(2S,4R)-1-((S)-2-(叔丁基)-21-((6-氯-8-氟-7-((R或S)-3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-18-甲基-4-氧代-6,9,12,15-四氧代-3,18-二氮杂苯三酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺19b。
步骤1
在零摄氏度,氮气保护搅拌条件下,向化合物三缩四乙二醇(20克,97.82毫摩尔,2当量)的二氯甲烷(300毫升)溶液中滴加三氟化硼的乙醚溶液(9.46毫克,0.06毫摩尔,0.01当量)和重氮 乙酸乙酯(5.88克,48.96毫摩尔,1.00当量)。在氮气保护搅拌条件下,混合物在25摄氏度下反应0.5小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%的甲醇/二氯甲烷流动相洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物19-1(黄色油,4.92克,产率35%)。MS(ESI,m/z):281.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.19(q,J=7.1Hz,2H),4.12(s,2H),3.75–3.61(m,14H),3.61–3.54(m,2H),1.26(t,J=7.1Hz,3H)。
步骤2
在零摄氏度,氮气保护搅拌条件下,向化合物19-1(4.00克,13.556毫摩尔,1.00当量)和N,N-二异丙基乙胺(2.95克,21.690毫摩尔,1.60当量)的二氯甲烷(30毫升)溶液中滴加甲基磺酰氯(2.12克,17.623毫摩尔,1.30当量)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物19-2(淡黄色油,4.15克,产率81%)。MS(ESI,m/z):359.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.41–4.34(m,2H),4.21(q,J=7.2Hz,2H),4.14(s,2H),3.80–3.60(m,14H),3.08(s,3H),1.28(t,J=7.1Hz,3H)。
步骤3
在25摄氏度搅拌条件下,向化合物19-2(4.15克,11.000毫摩尔,1.00当量)和3-(甲胺基)-1-丙醇(1.24克,13.200毫摩尔,1.20当量)的乙腈(50毫升)溶液中加入碳酸钾(3.20克,22.001毫摩尔,2.00当量)。混合物在80摄氏度下反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至室温。过滤除去不溶物,滤饼用二氯甲烷(50毫升 x 3)清洗。滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→13%甲醇(3%氨水)/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到粗产品。粗产品进一步通过反相快速色谱柱(C18柱)进行纯化,在15分钟内用5%→50%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器,UV205纳米;得到化合物19-3(淡黄色油,1.3克,产率31%)。MS(ESI,m/z):352.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.21(q,J=7.2Hz,2H),4.14(s,2H),3.82–3.75(m,2H),3.74–3.67(m,4H),3.67–3.57(m,10H),2.68–2.60(m,4H),2.32(s,3H),1.74–1.66(m,2H),1.28(t,J=7.1Hz,3H)。
步骤4
在25摄氏度,氮气保护搅拌条件下,向化合物1-7(910.00毫克,1.895毫摩尔,1.00当量)的 N-甲基吡咯烷酮(8毫升)溶液中加入氟化铯(431.83毫克,2.843毫摩尔,1.50当量),19-3(582.43毫克,1.65毫摩尔,1.0当量)和N,N-二异丙基乙胺(489.88毫克,3.790毫摩尔,2.00当量)。所得混合物在120摄氏度下搅拌反应3小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→100%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物19-4(棕色油,350毫克,产率22%)。MS(ESI,m/z):794.3/796.3/798.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.70(d,J=2.0Hz,1H),4.50(t,J=6.5Hz,2H),4.21(q,J=7.1Hz,2H),4.14(s,2H),3.81–3.59(m,22H),2.78–2.52(m,4H),2.42–2.26(m,3H),2.05(d,J=6.3Hz,2H),1.50(s,9H),1.28(t,J=7.2Hz,3H)。
步骤5
在25摄氏度搅拌条件下,向化合物19-4(310.00毫克,0.370毫摩尔,1.00当量)的水/甲醇/四氢呋喃(1/1/1,3毫升)溶液中加入氢氧化锂(18.67毫克,0.741毫摩尔,2当量)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入稀盐酸(1摩尔/升,88微升)中和。所得混合物减压浓缩得到粗产品酸。将该粗产品酸溶于N,N-二甲基甲酰胺(2.0毫升),然后加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(177.89毫克,0.444毫摩尔,1.2当量)。混合物在25摄氏度下反应1小时。向上述反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(190.66毫克,0.407毫摩尔,1.1当量)和N,N-二异丙基乙胺(201.55毫克,1.482毫摩尔,4.00当量)。混合物在25摄氏度下继续反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用30%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV 254纳米;得到化合物19-5(白色固体,300毫克,产率67%)。MS(ESI,m/z):1192.3/1194.3/1196.3[M+H]
+。
步骤6
在25摄氏度,氮气保护搅拌条件下,向化合物19-5(300.00毫克,0.251毫摩尔,1.00当量)的1,4-二氧六环/水(5/1,3.6毫升)的混合溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(101.84毫克,0.377毫摩尔,1.50当量),碳酸钠(53.28毫克,0.503毫摩尔,2.00当量)和[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(II)(20.47毫克,0.025毫摩尔,0.10当量)。反应液在 80摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物19-6(白色固体,280毫克,产率88%)。MS(ESI,m/z):1256.5/1258.6[M+H]
+。
步骤7
通过制备级手性高效液相色谱法对步骤6所得化合物19-6(280毫克)进行手性拆分:手性柱CHIRALPAK IC,2 x 25厘米,5微米;流动相A:正己烷/二氯甲烷=3/1(0.1%二乙基胺),流动相B:乙醇;流速:15毫升/分钟;在32分钟内用50%的B相进行洗脱,检测器UV220/254纳米。得到两个产品,较短保留时间(12.06分钟)的产品为化合物19-6a(类白色固体,130毫克,回收率48%),化合物19-6a:MS(ESI,m/z):1256.5/1258.6[M+H]
+;较长保留时间(23.776分钟)的产品为化合物19-6b(类白色固体,120毫克,回收率45%),化合物19-6b:MS(ESI,m/z):1256.5/1258.6[M+H]
+。
步骤8
在25摄氏度搅拌条件下,向化合物19-6a(60.00毫克,0.048毫摩尔,1.00当量)的二氯甲烷(1毫升)溶液中加入三氟乙酸(0.3毫升)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;梯度:在10分钟内用26%→55%B相梯度洗脱,检测器UV254/220纳米;得到19a(白色固体,23.8毫克,产率43%)。MS(ESI,m/z):1156.5/1158.5[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.04(s,1H),8.98(s,1H),8.43(d,J=7.6Hz,1H),7.91(d,J=1.6Hz,1H),7.80(d,J=8.4Hz,1H),7.47–7.40(m,3H),7.41–7.32(m,3H),7.28(d,J=2.4Hz,1H),7.25–7.19(m,2H),7.06(d,J=2.4Hz,1H),5.13(s,1H),4.94–4.87(m,1H),4.54(d,J=9.6Hz,1H),4.44(t,J=8.4Hz,1H),4.35(t,J=6.4Hz,2H),4.30–4.25(m,1H),3.94(s,2H),3.79–3.74(m,4H),3.63–3.52(m,6H),3.51–3.49(m,4H),3.48–3.44(m,6H),2.91–2.85(m,4H),2.48–2.46(m,4H),2.45(s,3H),2.18(s,3H), 2.09–2.00(m,1H),1.92–1.82(m,2H),1.81–1.73(m,1H),1.37(d,J=7.0Hz,3H),0.93(s,9H)。
步骤8’
在25摄氏度搅拌条件下,向化合物19-6b(60.00毫克,0.048毫摩尔,1.00当量)的二氯甲烷(1毫升)溶液中加入三氟乙酸(0.3毫升)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;梯度:在10分钟内用31%→65%的B相梯度洗脱,检测器:UV 254/220纳米;得到19b(白色固体,23.1毫克,产率41%)。MS(ESI,m/z):1156.5/1158.5[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.02(s,1H),8.98(s,1H),8.43(d,J=7.6Hz,1H),7.91(d,J=1.6Hz,1H),7.80(d,J=8.4Hz,1H),7.47–7.40(m,3H),7.40–7.32(m,3H),7.28(d,J=2.4Hz,1H),7.25–7.18(m,2H),7.06(d,J=2.4Hz,1H),5.20–5.09(m,1H),4.93–4.87(m,1H),4.54(d,J=9.6Hz,1H),4.48–4.40(m,1H),4.38–4.32(m,2H),4.30–4.26(m,1H),3.94(s,2H),3.84–3.70(m,4H),3.64–3.52(m,6H),3.52–3.49(m,4H),3.48–3.44(m,6H),2.92–2.84(m,4H),2.48–2.46(m,4H),2.45(s,3H),2.18(s,3H),2.06–2.02(m,1H),1.89–1.84(m,2H),1.81–1.73(m,1H),1.37(d,J=7.2Hz,3H),0.93(s,9H)。
实施例20
(2S,4R)-1-((S)-2-(叔丁基)-24-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-21-甲基-4-氧代-6,9,12,15,18-五氧代-3,21-二氮四甲酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺20a;(2S,4R)-1-((S)-2-(叔丁基)-24-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-21-甲基-4-氧代-6,9,12,15,18-五氧代-3,21-二氮四甲酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺20b。
步骤1
在零摄氏度搅拌条件下,向250毫升单口瓶中加入五甘醇(10.0克,41.5毫摩尔,1.0当量),三氟化硼的乙醚溶液(443.5毫克,3.1毫摩尔,0.1当量)和二氯甲烷(150.0毫升),然后缓慢滴加重氮乙酸乙酯(2.4克,20.7毫摩尔,0.5当量)。所得混合物在0摄氏度条件下搅拌反应2个小时。反应过程通过液质和薄层层析来监控。反应结束后,混合物恢复至25摄氏度,减压浓缩除去多余的试剂得到粗产品。所得粗产品通过硅胶柱层析进行纯化,用0%→4%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物20-1(黄色油状物,2.4克,产率18%)。MS(ESI,m/z):325.3[M+H]
+;
1HNMR(300MHz,CDCl
3)δ4.22(q,J=7.1Hz,2H),4.15(s,2H),3.78–3.69(m,6H),3.68–3.65(m,12H),3.63–3.59(m,2H),1.29(t,J=7.1Hz,3H)。
步骤2
在零摄氏度搅拌条件下,向20-1(2.4克,7.4毫摩尔,1.0当量)的二氯甲烷(50.0毫升)溶液中加入甲基磺酰氯(1.1克,9.3毫摩尔,1.3当量)和N,N-二异丙基乙胺(1.6克,11.4毫摩尔,1.6当量)。反应液在25摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩除去多余的试剂得到粗产品。所得粗产品通过硅胶柱层析进行纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物20-2(黄色油状物,2.0克,产率66%)。MS(ESI,m/z):403.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.44–4.34(m,2H),4.21(q,J=7.1Hz,2H),4.15(s,2H),3.82–3.59(m,18H),3.09(s,3H),1.29(t,J=7.1Hz,3H)。
步骤3
在25摄氏度搅拌条件下,向100毫升单口瓶中依次加入化合物20-2(2.0克,4.7毫摩尔,1.0当量),3-(甲胺基)-1-丙醇(0.5克,5.7毫摩尔,1.2当量),乙腈(50.0毫升)和碳酸钾(1.4克,9.4毫摩尔,2.0当量)。混合物在80摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至25摄氏度,减压浓缩除去多余的试剂得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→10%的甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物20-3(黄色油状物,1.2克,产率61%)。MS(ESI,m/z):396.2[M+H]
+;
1HNMR(400MHz,CDCl
3)δ4.22(q,J=7.1Hz,2H),4.15(s,2H),3.76–3.59(m,20H),2.70–2.60(m,4H),2.32(s,3H),1.75–1.66(m,2H),1.29(t,J=7.1Hz,3H)。
步骤4
在25摄氏度氮气保护条件下,向1-7(0.5克,0.9毫摩尔,1.0当量)的N-甲基吡咯烷酮(20.0毫升)的溶液中加入20-3(0.5克,1.2毫摩尔,1.3当量),氟化铯(0.2克,1.4毫摩尔,1.5当量)和入N,N-二异丙基乙胺(0.3克,1.9毫摩尔,2.0当量)。反应液在120摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至25摄氏度。减压浓缩除去多余的试剂得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→10%的甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物20-4(黄色油状物,463毫克,产率56%)。MS(ESI,m/z):838.1/840.1/842.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.72–7.69(m,1H),4.49(t,J=6.5Hz,2H),4.21(q,J=7.2Hz,2H),4.14(s,2H),3.80–3.71(m,6H),3.71–3.67(m,2H),3.66–3.58(m,18H),2.67(s,3H),2.50–2.25(m,4H),2.12–1.97(m,2H),1.49(s,9H),1.28(t,J=7.1Hz,3H)。
步骤5
在25摄氏度搅拌条件下,将20-4(160.0毫克,0.2毫摩尔,1.0当量)溶于四氢呋喃/甲醇/水(1/1/1,5.0毫升)的混合溶液中,再加入氢氧化锂(16.0毫克,0.4毫摩尔,2.0当量)。混合物在25摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,向反应液中加入1.0摩尔每升的稀盐酸调节pH至5。减压浓缩得到中间体。然后在25摄氏度搅拌条件下,将中间体溶于N,N-二甲基甲酰胺(5.0毫升)中,再加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(90.0毫克,0.24毫摩尔,1.2当量)。反应液在25摄氏度条件下反应15分钟,再加入N,N-二异丙基乙胺(91.8毫克,0.7毫摩尔,3.6当量)和(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(105.2毫克,0.2毫摩尔,1.2当量)。反应液在25摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后。减压浓缩得到粗产品。粗产品通过反相色谱柱(C18柱)进行纯化,在25分钟内用30%→95%的乙腈/水流动相(0.1%碳酸氢氨)进行洗脱;检测器,UV254/220纳米;得到化合物20-5(白色固体,180.0毫克,产率76%)。MS(ESI,m/z):1236.3/1238.3/1240.3[M+H]
+。
步骤6
在25摄氏度氮气保护条件下,向化合物20-5(162.3毫克,0.1毫摩尔,1.0当量)的1,4-二氧六环/水(1/1,6.0毫升)混合溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(53.1毫克,0.2毫摩尔,1.5当量),[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(10.7毫克,0.01毫摩尔,0.1当量)和碳酸钠(27.8毫克,0.3毫摩尔,2.0当量)。混合物在90摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至室温,减压浓缩得到粗产品。粗产品通过反相色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV 254/220纳米;得到化合物20-6(白色固体,140.0毫克,产率81%)。MS(ESI,m/z):1300.5/1302.5[M+H]
+;
1HNMR(400MHz,CDCl
3)δ8.67(s,1H),7.80–7.81(m,2H),7.46–7.27(m,9H),7.25–7.19(m,1H),7.17–7.13(m,1H),5.13–5.03(m,1H),4.79–4.68(m,1H),4.63–4.54(m,2H),4.12–3.94(m,3H),3.91–3.76(m,3H),3.76–3.52(m,22H),3.49(s,9H),2.96–2.66(m,3H),2.51(s,3H),2.49–2.36(m,2H),2.20–2.06(m,3H),1.51(s,9H),1.48–1.45(m,3H),1.08–1.02(m,8H)。
步骤7
通过制备级手性高效液相色谱法对步骤6所得化合物20-6(140毫克)进行手性拆分:手性柱NB-Lux 5 m i-Cellulose-5,2.12 x 25厘米,5微米;流动相A:正己烷/二氯甲烷=3/1(0.1%二乙胺),流动相B:乙醇;流速:15毫升/分钟;用50%流动相B洗脱28.2分钟;检测器UV220/254纳米,得到两个产品。较短保留时间(11.2分钟)的产品为化合物20-6a(白色固体,60.0毫克,回收率43%),化合物20-6a:MS(ESI,m/z):1300.5/1302.5[M+H]
+;较长保留时间(20.6分钟)的产品为化合物20-6b(白色固体,61.0毫克,回收率44%),化合物20-6b:MS(ESI,m/z):1300.5/1302.5[M+H]
+。
步骤8
在零摄氏度搅拌条件下,向化合物20-6a(60.0毫克,0.05毫摩尔,1.0当量)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(1.0毫升)。混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米;流动相A:水(0.1%碳酸氢氨),流动相B:乙腈;流速:25毫升/分钟;梯度:在2分钟内用10%的B相洗脱,然后在2.5分钟内用10%→28%的B相梯度洗脱,最后在11.5分钟内用28%→58%的B相梯度洗脱;检测器UV 254/220纳米;得到化合物20a(白色固体,23.0毫克,产率42%)。MS(ESI,m/z):1200.4/1202.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.02(s,1H),8.98(s,1H),8.44(d,J=7.7Hz,1H),7.94–7.90(m,1H),7.83–7.78(m,1H),7.49–7.40(m,3H),7.40–7.32(m,3H),7.28(d,J=2.4Hz,1H),7.24–7.18(m,2H),7.08–7.04(m,1H),5.20–5.10(m,1H),4.96–4.86(m,1H),4.54(d,J=9.5Hz,1H),4.48–4.40(m,1H),4.39–4.32(m,2H),4.31–4.25(m,1H),3.97–3.93(m,2H),3.81–3.73(m,4H),3.65–3.42(m,21H),2.94–2.84(m,4H),2.49–2.46(m,3H),2.45(s,3H),2.18(s,3H),2.09–2.00(m,1H),1.92–1.82(m,2H),1.81–1.72(m,1H),1.42–1.32(m,3H),0.93(s,9H)。
步骤8’
在零摄氏度搅拌条件下,向化合物20-6b(61.0毫克,0.05毫摩尔,1.0当量)的二氯甲烷(2.0毫升)溶液中加入三氟乙酸(1.0毫升)。混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米;流动相A:水(0.1%碳酸氢氨),流动相B:乙腈;流速:25毫升/分钟;梯度:在10分钟内用26%→55%的B相梯度洗脱;检测器UV 254/220纳米;得到20b(白色固体,24.0毫克,产率42%)。MS(ESI,m/z):1200.4/1202.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.02(s,1H),8.98(s,1H),8.44(d,J=7.7Hz,1H),7.94–7.90(m,1H),7.83–7.78(m,1H),7.49–7.40(m,3H),7.40–7.32(m,3H),7.28(d,J=2.4Hz,1H),7.24–7.18(m,2H),7.08–7.04(m,1H),5.20–5.10(m,1H),4.96–4.86(m,1H),4.54(d,J=9.5Hz,1H),4.48–4.40(m,1H),4.39–4.32(m,2H),4.30–4.25(m,1H),3.97–3.93(m,2H),3.82–3.74(m,4H),3.65–3.42(m,21H),2.94–2.86(m,4H),2.49–2.46(m,3H),2.45(s,3H),2.19(s,3H),2.09–2.00(m,1H),1.93–1.82 (m,2H),1.81–1.72(m,1H),1.41–1.33(m,3H),0.93(s,9H)。
实施例21
(2S,4R)-1-((S)-18-(叔丁基)-2-(1-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氮杂环丁烷-3-基)-16-氧代-5,8,11,14-四氧代-2,17-二氮杂环癸烷-19-酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺21a;(2S,4R)-1-((S)-18-(叔丁基)-2-(1-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氮杂环丁烷-3-基)-16-氧代-5,8,11,14-四氧代-2,17-二氮杂环癸烷-19-酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺21b。
步骤1
在25摄氏度搅拌条件下,向化合物3-(甲基氨基)氮杂丁烷-1-羧酸叔丁酯(3.00克,15.302毫摩尔,1.00当量)的乙腈(30毫升)溶液中加入N,N-二异丙基乙胺(3.96克,30.604毫摩尔,2.00当量)和氯丙烯(1.41克,18.362毫摩尔,1.20当量)。反应液在80摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至室温。减压浓缩除去溶剂,加入水(30毫升),然后用乙酸乙酯(30毫升 x 3)萃取。合并后有机层用30毫升饱和氯化钠溶液洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩得到粗产品。粗产品用0%→25%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物21-1(红色油,3克,产率86%)。MS(ESI,m/z):227.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ6.37–6.23(m,1H),5.71–5.60(m,2H),4.43–4.34(m,2H),4.34–4.24(m,2H),3.74–3.62(m,1H),3.43–3.37(m,2H),2.61(s,3H),1.90(s,9H)。
步骤2
在25摄氏度搅拌条件下,向化合物21-1(424.21毫克,1.781毫摩尔,1.50当量)的二氯甲烷(3毫升)溶液中加入三氟乙酸(1毫升)。反应液在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液减压浓缩得中间体粗产品。将该中间体溶于N-甲基吡咯烷酮(5毫升),随后向其中加入1-7(600.00毫克,1.187毫摩尔,1.00当量)和N,N-二异丙基乙胺(1.62克,11.870毫摩尔,10.00当量)。所得混合物在60摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→95%的乙腈/水流动相(0.1%甲酸)进行洗脱;检测器UV254纳米;得到化合物21-2(淡黄色固体,650毫克,产率96%)。MS(ESI,m/z):569.3/571.3/573.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.56(d,J=1.9Hz,1H),5.95–5.81(m,1H),5.29–5.15(m,2H),4.31–4.18(m,2H),4.16–4.03(m,2H),3.62(s,8H),3.49–3.37(m,1H),3.10–2.95(m,2H),2.24(s,3H),1.49(s,9H)。
步骤3
在25摄氏度搅拌条件下,向化合物21-2(570.0毫克,0.95毫摩尔,1.00当量)的二氯甲烷(10毫升)溶液中加入1,3-二甲基巴比妥酸(234.3毫克,1.43毫摩尔,1.50当量),四(三苯基膦)钯(57.8毫克,0.048毫摩尔,0.05当量)。混合物在25摄氏度条件下搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物21-3(淡黄色固体,350毫克,产率88%)。MS(ESI,m/z):529.1/531.1/532.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.57–7.54(m,1H),4.42–4.33(m,2H),3.98–3.86(m,2H),3.74–3.68(m,1H),3.62(s,8H),2.46(s,3H),1.49(s,9H)。
步骤4
在零摄氏度搅拌条件下,向乙酸叔丁酯-四聚乙二醇(623毫克,2.0毫摩尔,1当量)的二氯甲烷(5.0毫升)溶液中滴加三氟乙酸(5.0毫升)。所得混合物在25摄氏度下搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得粗产品。向粗产品中加入甲苯(15.0毫升),减压浓缩进一步除去残余的三氟乙酸,重复此操作3次得到化合物21-4的粗产品(淡黄色油,490毫克)。粗产品直接用于下一步反应。MS(ESI,m/z):253.1[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向化合物21-4(490.00毫克,1.9毫摩尔,1.00当量)的N,N-二甲基甲酰胺(5.0毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(887.2毫克,2.3毫摩尔,1.20当量)。混合物在25摄氏度下反应0.5小时。然后向上述反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(1.03克,2.1毫摩尔,1.10当量)和N,N-二异丙基乙胺(1.23克,9.5毫摩尔,5.00当量)。混合物在25摄氏度下继续反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液通过反相快速色谱柱(C18柱)进行纯化,在14分钟内用5%→95%的乙腈/水流动相进行洗脱;检测器UV254纳米;得到化合物21-5(淡黄色固体,785毫克,产率60%)。MS(ESI,m/z):679.3[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ8.99(s,1H),8.45(d,J=7.7Hz,1H),7.46–7.36(m,5H),4.96–4.85(m,1H),4.59–4.53(m,1H),4.49–4.42(m,1H),4.32–4.27(m,1H),3.99–3.94(m,2H),3.65–3.47(m,16H),3.44–3.40(m,2H),2.46(s,3H),2.11–2.01(m,1H),1.84–1.73(m,1H),1.46–1.33(m,3H),0.95(s,9H)。
步骤6
在25摄氏度搅拌条件下,向化合物21-5(785.00毫克,1.16毫摩尔,1.00当量)的乙腈(8.0毫升)溶液中加入2-碘酰基苯甲酸(356.6毫克,1.27毫摩尔,1.10当量)。混合物在60摄氏度下反应2.5小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→8%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物21-6(白色固体,615.3毫克,产率78%)。MS(ESI,m/z):677.2[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物21-6(332.13毫克,0.49毫摩尔,1.30当量)和化合物21-2(200.00毫克,0.38毫摩尔,1.00当量)的甲醇(8毫升)溶液中加入乙酸(22.67毫克,0.38毫摩尔,1.00当量)和氰基硼氢化钠(35.58毫克,0.57毫摩尔,1.50当量)。混合物在25摄氏度下反应 3小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用制备薄层层析法纯化(二氯甲烷/甲醇=12/1),得到化合物21-7(白色固体,356毫克,产率78%)。MS(ESI,m/z):1189.4/1191.3/1193.4[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ8.98(s,1H),8.43(d,J=7.8Hz,1H),7.77(s,1H),7.46–7.34(m,5H),5.13(d,J=3.5Hz,1H),4.96–4.85(m,1H),4.54(d,J=9.5Hz,1H),4.44(t,J=8.1Hz,1H),4.33–4.24(m,1H),4.19–4.07(m,2H),3.99–3.93(m,2H),3.92–3.84(m,2H),3.71–3.46(m,26H),2.45(s,3H),2.26–2.15(m,3H),2.11–2.01(m,1H),1.84–1.71(m,1H),1.43(s,9H),1.37(d,J=6.9Hz,3H),0.94(s,9H)。
步骤8
在25摄氏度,氮气保护搅拌条件下,向化合物21-7(200.00毫克,0.160毫摩尔,1.00当量)的四氢呋喃/水(10/1,2.2毫升)溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(68.06毫克,0.252毫摩尔,1.5当量),磷酸钾(71.31毫克,0.336毫摩尔,2.0当量)和氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(14.22毫克,0.017毫摩尔,0.1当量)。反应液在60摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至25摄氏度,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物21-8(类白色固体,170毫克,产率84%)。MS(ESI,m/z):1253.7/1255.7[M+H]
+。
步骤9
通过制备级手性高效液相色谱法对步骤8所得化合物20-8(170毫克)进行手性拆分:手性柱NB-Lux 5 m i-Cellulose-5,2.12 x 25厘米,5微米;流动相A:正己烷/甲基叔丁基醚=1/1(0.5%,2摩尔/升氨甲醇),流动相B:乙醇;流速:20毫升/分钟;在8分钟内用50%的B相进行洗脱,检测器UV225/250纳米,得到两个产品。较短保留时间(8.645分钟)的产品为化合物21-8a(类白色固体,50毫克,回收率29%),化合物21-8a:MS(ESI,m/z):1253.7/1255.7[M+H]
+;较长保留时间(15.475分钟)的产品为化合物21-8b(类白色固体,50毫克,回收率29%),化合物21-8b:MS(ESI,m/z): 1253.7/1255.7[M+H]
+。
步骤10
在25摄氏度搅拌条件下,向化合物21-8a(50.00毫克,0.040毫摩尔,1.00当量)的二氯甲烷(3毫升)溶液中加入三氟乙酸(1毫升)。混合物在25摄氏度下反应0.5小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度在7分钟内用5%→25%的B相梯度洗脱;检测器UV254/220纳米;得到21a(类白色固体,10毫克,产率21%)。MS(ESI,m/z):1153.5/1155.4[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.90–8.86(m,1H),7.81–7.73(m,2H),7.47–7.38(m,5H),7.30–7.17(m,3H),7.03(d,J=2.4Hz,1H),5.06–4.97(m,1H),4.69(s,1H),4.63–4.54(m,1H),4.44(s,1H),4.34–4.22(m,2H),4.14–4.02(m,4H),3.91–3.43(m,22H),3.14–3.04(m,4H),2.71–2.62(m,2H),2.48(s,3H),2.32(s,3H),2.26–2.14(m,1H),2.02–1.91(m,1H),1.60–1.48(m,3H),1.40–1.26(m,2H),1.04(s,9H),0.96–0.84(m,1H)。
步骤10’
在25摄氏度搅拌条件下,向化合物21-8b(50.00毫克,0.040毫摩尔,1.00当量,)的二氯甲烷(3毫升)溶液中加入三氟乙酸(1毫升)。混合物在25摄氏度下反应0.5小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度在7分钟内用5%→25%B相梯度洗脱;检测器UV 254/220纳米;得到21b(类白色固体,12毫克,产率25%)。MS(ESI,m/z):1153.5/1155.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.00(s,1H),8.98(s,1H),8.44(d,J=7.7Hz,1H),7.79(d,J=8.3Hz,1H),7.74(d,J=1.6Hz,1H),7.48–7.40(m,3H),7.40–7.32(m,3H),7.26(d,J=2.4Hz,1H),7.21(d,J=4.0Hz,2H),7.03(d,J=2.3Hz,1H),5.22–5.08(m,1H),4.97–4.83(m,1H),4.54(d,J=9.5Hz,1H),4.50–4.39(m,1H),4.34–4.22(m,1H),4.18–4.05(m,2H),3.98–3.83(m,4H),3.72–3.63(m,4H),3.62–3.44(m,18H),2.96–2.88(m,4H),2.45(s,3H),2.18(s,3H),2.09–1.99(m,1H),1.82–1.73(m,1H),1.37(d,J=7.0Hz,3H),1.29–1.21(m,1H),0.93(s,9H)。
实施例22
(2S,4R)-1-((S)-2-(5-(4-(5-((3-((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)戊基)哌嗪-1-基)戊酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺三盐酸盐22。
步骤1
在25摄氏度搅拌条件下,向100毫升圆底烧瓶中依次加入5-溴戊酸甲酯(2.0克,9.7毫摩尔,1.0当量),乙腈(30.0毫升),1-叔丁氧羰基哌嗪(2.1克,10.7毫摩尔,1.1当量)和碳酸钾(2.8克,19.4毫摩尔,2.0当量)。所得混合物在85摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,过滤,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物22-1(黄色液体,2.8克,产率91%)。MS(ESI,m/z):301.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ3.68(s,3H),3.47–3.40(m,4H),2.40–2.31(m,8H),1.73–1.60(m,2H),1.56–1.52(m,2H),1.47(s,9H)。
步骤2
在25摄氏度搅拌条件下,向化合物22-1(1.4克,4.7毫摩尔,1.0当量)的甲醇/四氢呋喃/水(1/1/1,13.5毫升)溶液中加入一水合氢氧化锂(0.3克,7.0毫摩尔,1.5当量)。所得混合物在25摄氏度下搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,在零摄氏度下向反应液中滴加盐酸溶液(1摩尔每升,7毫升)。反应液浓缩后得到粗产品22-2(白色固体,1.8克)。该粗产 品直接用于下一步合成。MS(ESI,m/z):287.2[M+H]
+。
步骤3
在25摄氏度搅拌条件下,向化合物22-2(472.2毫克)的N,N-二甲基甲酰胺(5.0毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(513.1毫克,1.3毫摩尔,1.2当量)。所得混合物在25摄氏度下搅拌反应15分钟。然后向反应体系中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(500.0毫克,1.1毫摩尔,1.0当量)和N,N-二异丙基乙胺(0.6毫升,3.3毫摩尔,3当量)。所得混合物在25摄氏度下继续搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用25%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物22-3(白色固体,640毫克,产率83%)。MS(ESI,m/z):713.4[M+H]
+。
步骤4
在25摄氏度搅拌条件下,向化合物22-3(300.0毫克,0.4毫摩尔,1.0当量)的二氯甲烷(6.0毫升)溶液中加入三氟乙酸(2.0毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品22-4(黄色固体,410.0毫克)。该化合物不需进一步纯化,直接用于下一步合成。MS(ESI,m/z):613.2[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向化合物22-4(72.9毫克,0.1毫摩尔,1.2当量),醋酸钠(12.2毫克, 0.15毫摩尔,1.5当量),醋酸(8.9毫克,0.15毫摩尔,1.5当量)和氰基硼氢化钠(18.7毫克,0.3毫摩尔,3.0当量)的甲醇(2.0毫升)溶液中滴加化合物3-6a(70.0毫克,0.1毫摩尔,1.0当量)。所得混合物在25摄氏度下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物22-5a(淡黄色固体,70毫克,产率54%)。MS(ESI,m/z):1302.8/1304.8[M+H]
+。
步骤6
在25摄氏度搅拌条件下,向化合物22-5a(50.0毫克,0.04毫摩尔,1.0当量)的甲醇(2.0毫升)溶液中加入盐酸的1,4-二氧六环溶液(4摩尔/升,2.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.05%盐酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→28%的B相梯度洗脱;检测器UV 254/220纳米;得到化合物22(黄色固体,21.5毫克,产率45%)。MS(ESI,m/z):1202.6/1208.6[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ12.00(s,2H),10.79(s,1H),10.08(d,J=10.0Hz,1H),9.84(s,1H),9.10(s,1H),8.46(d,J=7.8Hz,1H),8.00(d,J=1.7Hz,1H),7.91(d,J=9.0Hz,1H),7.82(d,J=8.3Hz,1H),7.54–7.35(m,5H),7.32(d,J=2.4Hz,1H),7.22(d,J=6.1Hz,2H),7.12(d,J=2.3Hz,1H),4.92(t,J=7.2Hz,2H),4.52–4.40(m,5H),4.29(s,1H),4.17(s,2H),3.99–3.83(m,2H),3.75–3.72(m,3H),3.63–3.58(m,3H),3.49(d,J=9.6Hz,4H),3.27–3.23(m,2H),3.15–3.05(m,6H),2.75(d,J=4.8Hz,3H),2.47(s,3H),2.25(d,J=9.1Hz,5H),1.98–1.92(m,5H),1.79–1.71(m,7H),1.56(d,J=7.3Hz,2H),1.39–1.30(m,5H),0.95(s,9H);
19F NMR(282MHz,DMSO-d
6)δ-122.00。
实施例23
(2S,4R)-1-((S)-2-(14-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)十四酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺23a;(2S,4R)-1-((S)-2-(14-((3-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)十四酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺23b。
步骤1
在25摄氏度搅拌条件下,向14-羟基肉豆蔻酸(200.0毫克,0.818毫摩尔,1.0当量)的N,N-二甲基甲酰胺(5毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(373.42毫克,0.982毫摩尔,1.2当量)。所得混合物在25摄氏度反应30分钟。然后向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(363.86毫克,0.818毫摩尔,1.0当量)和N,N-二异丙基乙胺(528.42毫克,4.090毫摩尔,5.0当量)。所得混合物在25摄氏度继续反应2.5小时。反应过程通过液质和薄层层析来监控。反应完成后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物23-1(白色固体,320毫克,产率58%)。MS(ESI,m/z):669.4[M-H]
-;
1H NMR(300MHz,DMSO-d
6)δ8.99(s,1H),8.38(d,J=7.8Hz,1H),7.79(d,J=9.3Hz,1H),7.47–7.35(m,4H),5.15–5.08(m,1H),5.00–4.85(m,1H),4.52(d,J=9.3Hz,1H),4.48–4.38(m,1H),4.38–4.22(m,2H),3.66–3.55(m,2H),2.46(s,3H),2.31–2.19(m,1H),2.15–1.95(m,2H),1.85–1.74(m,1H),1.51–1.35(m,7H),1.29–1.20(m,19H),0.93(s,9H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向反应瓶中依次加入化合物23-1(300毫克,0.425毫摩尔,1.0当量),2-碘酰基苯甲酸(125.2毫克,0.425毫摩尔,1.0当量)和乙腈(6毫升)。所得混合物在60摄氏度氮气保护下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→12%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物23-2(白色固体,174毫克,产率58%)。MS(ESI,m/z):669.4[M+H]
+。
步骤3
在零摄氏度氮气保护搅拌条件下,向25毫升单口瓶中依次加入2-9a(30毫克,0.048毫摩尔,1.0当量),23-2(37毫克,0.055毫摩尔,1.1当量),醋酸(4.53毫克,0.075毫摩尔,1.5当量),甲醇(4毫升)和氰基硼氢化钠(4.74毫克,0.075毫摩尔,1.5当量)。所得混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完成后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物23-3a(白色固体,44.5毫克,产率74%)。MS(ESI,m/z):1248.6/1250.6[M+H]
+。
步骤4
在零摄氏度搅拌条件下,向25毫升单口瓶中依次加入化合物23-3a(44.5毫克,0.035毫摩尔,1.0当量),二氯甲烷(4毫升)和三氟乙酸(2毫升)。反应液在20摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液浓缩得到粗产品,粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.05%氨水)进行洗脱;检测器UV 254纳米;得到化合物23a(白色固体,5毫克,产率12%)。MS(ESI,m/z):1148.6/1150.6[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.87(s,1H),7.95–7.92(m,1H),7.74(d,J=8.2Hz,1H),7.45–7.36(m,5H),7.27–7.16(m,3H),7.04–7.01(m,1H),5.04–4.95(m,1H),4.61(s,1H),4.59–4.53(m,1H),4.53–4.47(m,2H),4.44–4.39(m,1H),3.96–3.85(m,5H),3.77–3.71(m,1H),3.10–3.02(m,4H),2.77–2.68(m,2H),2.54–2.45(m,5H),2.37(s,3H),2.32–2.15(m,3H),2.09–2.01(m,2H),1.99–1.90(m,1H),1.62–1.47(m,7H),1.37–1.33(m,1H),1.30–1.21(m,16H),1.06–0.99(m,9H),0.92–0.81(m,1H)。
步骤3’
在零摄氏度氮气保护搅拌条件下,向25毫升单口瓶中依次加入2-9b(40毫克,0.067毫摩尔,1.0当量),23-2(58毫克,0.087毫摩尔,1.3当量),醋酸(6.04毫克,0.101毫摩尔,1.5当量),甲醇(5毫升)和氰基硼氢化钠(6.33毫克,0.101毫摩尔,1.5当量)。所得混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完成后,减压浓缩得到粗产品。粗产品用硅胶柱 层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物23-3b(白色固体,71毫克,产率85%)。MS(ESI,m/z):1248.6/1250.6[M+H]
+。
步骤4’
在零摄氏度搅拌条件下,向25毫升单口瓶中依次加入23-3b(71毫克,0.058毫摩尔,1.0当量),二氯甲烷(4毫升)和三氟乙酸(2毫升)。反应液在20摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液减压浓缩的粗产品,粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.05%氨水)进行洗脱;检测器UV 254纳米;得到化合物23b(白色固体,15毫克,产率23%)。MS(ESI,m/z):1148.6/1150.6[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.86(s,1H),7.95–7.90(m,1H),7.74(d,J=8.3Hz,1H),7.47–7.33(m,5H),7.26–7.15(m,3H),7.05–7.01(m,1H),5.03–4.95(m,1H),4.61(s,1H),4.58–4.53(m,1H),4.53–4.47(m,2H),4.44–4.39(m,1H),3.95–3.84(m,5H),3.77–3.70(m,1H),3.09–3.01(m,4H),2.73–2.63(m,2H),2.51–2.42(m,5H),2.34–2.30(m,3H),2.29–2.15(m,3H),2.08–1.99(m,2H),1.98–1.89(m,1H),1.64–1.45(m,7H),1.37–1.33(m,1H),1.30–1.21(m,16H),1.02(d,J=11.0Hz,9H),0.94–0.83(m,1H)。
实施例24
(2S,4R)-1-((S)-2-(4-(4-(4-((3-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁基)哌嗪-1-基)丁酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺二甲酸盐24。
步骤1
在25摄氏度搅拌条件下,向4-氯丁酸甲酯(1.10克,8.1毫摩尔,1.00当量)的乙腈(10毫升)溶液中加入1-叔丁氧羰基哌嗪(1.50克,8.1毫摩尔,1.00当量)和碳酸钾(2.23克,16.2毫摩尔,2.00当量)。反应液在80摄氏度条件下搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应完全后,反应液减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物24-1(白色固体,623.4毫克,产率26%)。MS(ESI,m/z):287.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ3.69(s,3H),3.43(t,J=5.1Hz,4H),2.43–2.34(m,8H),1.90–1.78(m,2H),1.48(s,9H)。
步骤2
在25摄氏度搅拌条件下,向化合物24-1(619.0毫克,2.16毫摩尔,1.0当量)的甲醇/四氢呋喃/水(1/1/1,7.5毫升)溶液中加入一水合氢氧化锂(136.0毫克,3.24毫摩尔,1.5当量)。所得混合物在25摄氏度下搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,在零摄氏度下向反应液中滴加盐酸溶液(1摩尔每升,3.3毫升)。反应液浓缩后得到粗产品24-2(白色固体,938毫克,粗产品)。粗产品直接用于下一步合成。MS(ESI,m/z):273.2[M+H]
+。
步骤3
在25摄氏度搅拌条件下,向24-2(320.00毫克,粗品)的N,N-二甲基甲酰胺溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(491.44毫克,1.228毫摩尔,1.10当量)。混合物在25摄氏度下搅拌1小时。然后向反应液中加入N,N-二异丙基乙胺(607.43毫克,4.465毫摩尔,4.00当量)和(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(10.45毫克,0.022毫摩尔,1.1当量)。混合物在25摄氏度下继续搅拌2小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液直接通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→70%的乙腈/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物24-3(亮黄色固体,560毫克,产率71%)。MS(ESI,m/z):699.5[M+H]
+。
步骤4
在零摄氏度搅拌条件下,向8毫升反应瓶中依次加入24-3(50.0毫克,0.068毫摩尔,1.00当量),乙酸乙酯(1.00毫升),盐酸的1,4-二氧六环溶液(4摩尔/升,1.00毫升)。所得混合物在25摄氏度搅拌1小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到化合物24-4的粗产品(亮黄色固体,40毫克,粗产品)。粗产品直接用于下一步反应。MS(ESI,m/z):599.3[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向8毫升反应瓶中依次加入化合物24-4(30.56毫克,0.048毫摩尔,0.85当量),甲醇(2.00毫升,62.418毫摩尔,822.71当量),醋酸(7.21毫克,0.114毫摩尔,2.00当量),氰基硼氢化钠(7.55毫克,0.114毫摩尔,2.00当量)和化合物8-6a(40.00毫克,0.057毫摩尔,1.00当量)。混合物在25摄氏度搅拌下反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物24-5a(白色固体,50毫克,产率70%)。MS(ESI,m/z):1148.6/1150.6[M+H]
+。
步骤6
在零摄氏度搅拌条件下,向8毫升反应瓶中依次加入化合物24-5a(25.0毫克,0.019毫摩尔,1.0当量),二氯甲烷(4.0毫升)和三氟乙酸(1.0毫升)。所得混合物在25摄氏度条件下搅拌1小时, 反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%甲酸)进行洗脱;检测器UV254纳米;得到化合物24(白色固体,7.2毫克,产率30%)。MS(ESI,m/z):1149.6[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.77(s,1H),8.33(s,2H),7.92(d,J=1.7Hz,1H),7.69–7.64(m,1H),7.35–7.24(m,5H),7.18(d,J=2.4Hz,1H),7.13–7.07(m,2H),6.93(d,J=2.4Hz,1H),4.94–4.86(m,1H),4.73–4.65(m,1H),4.55–4.41(m,4H),4.37–4.31(m,1H),4.05–3.93(m,4H),3.81–3.75(m,1H),3.68–3.61(m,1H),3.35–3.26(m,4H),3.24–3.22(m,1H),3.19–3.14(m,1H),3.06–2.98(m,2H),2.81–2.63(m,10H),2.61–2.55(m,2H),2.53–2.47(m,2H),2.39–2.35(m,3H),2.31–2.25(m,2H),2.22–2.07(m,3H),1.90–1.81(m,1H),1.81–1.60(m,4H),1.57–1.50(m,2H),1.47–1.37(m,3H),0.93(d,J=9.5Hz,9H);
19F NMR(377MHz,CD
3OD)δ-123.10。
实施例25
(2S,4R)-1-((S)-2-(3-(3-((S)-2-((((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丙氧基)丙胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐25。
步骤1
在零摄氏度,氮气保护搅拌条件下,向3-丁烯-1-醇(10.0克,131.7毫摩尔,1.0当量),三乙胺(28.0克,263.55毫摩尔,2.0当量)和4-二甲氨基吡啶(2.5克,19.7毫摩尔,0.15当量)的二氯甲烷(100毫升)溶液中滴加对甲基苯磺酰氯(29.1克,144.9毫摩尔,1.1当量)。滴加完毕后,混合物在25摄氏度氮气保护搅拌条件下反应16小时。反应过程通过液质和薄层层析来监控。反应完全后,向体系中加入200毫升水淬灭反应。混合物用二氯甲烷(200毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→25%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物25-1(无色油状,29.00克,产率92.4%)。MS(ESI,m/z):227.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.82–7.76(m,2H),7.35(d,J=8.0Hz,2H),5.72–5.62(m,1H),5.10–5.05(m,2H),4.08–4.05(m,2H),2.45(s,3H),2.43–2.37(m,2H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向100毫升单口瓶中依次将化合物25-1(10.0克,132.6毫摩尔,2.0当量)和氢氧化钾(7.44克,132.6毫摩尔,2.0当量),将混合物加热到75摄氏度反应30分钟后。然后在75摄氏度将化合物1,3-丙二醇(15.0克,62.9毫摩尔,1.0当量)缓慢滴加至上述体系中,滴加时间不低于30分钟,滴加完毕后将反应体系加热至80摄氏度反应30分钟。反应通过薄层色谱监控,反应完毕后将反应体系冷却至25摄氏度。反应体系用200毫升水淬灭,用乙酸乙酯(200毫升 x 3)萃取,合并有机相,用无水硫酸钠干燥后过滤,所得滤液通过减压蒸馏除去溶剂。流动相用0%→25%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物25-2(无色油状,2.15克,产率25%)。
1H NMR(400MHz,CDCl
3)δ5.86–5.76(m,1H),5.13–5.04(m,2H),3.78–3.75(m,2H),3.65–3.62(m,2H),3.52–3.48(m,2H),2.56(s,1H),2.37–2.31(m,2H),1.86–1.80(m,2H)。
步骤3
在零摄氏度,氮气保护搅拌条件下,向化合物25-2(2.15克,16.5毫摩尔,1.0当量),三乙胺(3.5克,33.1毫摩尔,2.0当量)和4-二甲氨基吡啶(320毫克,2.5毫摩尔,0.15当量)的二氯甲烷(20毫升)溶液中滴加对甲基苯磺酰氯(3.46克,18.1毫摩尔,1.1当量)。滴加完毕后,混合物在25摄氏度氮气保护搅拌条件下反应16小时。反应过程通过液质和薄层层析来监控。反应完全后,向体系中加入20毫升水淬灭反应。混合物用二氯甲烷(20毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→25%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物25-3(无色油状,4.0克,产率89%)。MS(ESI,m/z):285.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.81–7.78(m,2H),7.35(d,J=8.0Hz,2H),5.79–5.69(m,1H),5.07–4.99(m,2H),4.15–4.11(m,2H),3.45–3.42(m,2H),3.38–3.35(m,2H),2.45(s,3H),2.26–2.20(m,2H),1.92–1.86(m,2H)。
步骤4
在25摄氏度搅拌条件下,向100毫升单口瓶中依次加入化合物25-3,L-脯氨醇(0.85克,7.9毫摩尔,1.2当量),碳酸钾(1.94克,13.3毫摩尔,2.0当量)和乙腈(20.0毫升)。混合物在50摄氏 度反应4小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,过滤除去不溶物,滤液通过减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物25-4(白色固体,1.17克,产率77%)。MS(ESI,m/z):214.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ5.90–5.77(m,1H),5.14–5.02(m,2H),3.66–3.61(m,1H),3.54–3.45(m,4H),3.41–3.37(m,1H),3.22–3.16(m,1H),3.05(s,1H),2.93–2.83(m,1H),2.64–2.58(m,1H),2.39–2.22(m,4H),1.93–1.70(m,6H)。
步骤5
在零摄氏度,氮气保护搅拌条件下,向化合物16-3a(300.0毫克,0.465毫摩尔,1.0当量)和25-4(125.1毫克,0.557毫摩尔1.2当量)的四氢呋喃(5.0毫升)溶液中缓慢滴加叔丁醇钾(65.8毫克,0.557毫摩尔,1.2当量)。混合物在零摄氏度条件下反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,将混合物倒入20毫升水中淬灭。混合物用乙酸乙酯(20毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物25-5a(白色固体,240毫克,产率62%)。MS(ESI,m/z):790.4/792.4[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.84–7.80(m,2H),7.54–7.46(m,2H),7.35–7.30(m,2H),7.19(d,J=2.4Hz,1H),5.71(s,1H),5.36–5.32(m,2H),5.03–4.96(m,2H),4.41(s,4H),3.95–2.92(m,15H),2.31–1.85(m,13H),1.53(s,9H)。
步骤6
在25摄氏度搅拌条件下,向25毫升单口瓶中依次加入25-5a(150.0毫克,0.18毫摩尔,1.0当量),高碘酸钠(166.43毫克,0.739毫摩尔,4.1当量),三氯化钌(4.28毫克,0.018毫摩尔,0.1当量),四氯化碳(1.0毫升),乙腈(1毫升)和水(1.5毫升)。混合物在25摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,向反应液中加入10毫升水稀释,用氯仿/异丙醇(3/1,10毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水 流动相(0.1%碳酸氢铵)进行洗脱;检测器UV220纳米;得到化合物25-6a(白色固体,78毫克,产率42%)。MS(ESI,m/z):808.4/810.4[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向8毫升反应瓶中依次加入25-6a(65.0毫克,0.064毫摩尔,1.0当量),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(23.3毫克,0.077毫摩尔,1.2当量)和N,N-二甲基甲酰胺(1毫升)。混合物在25摄氏度条件下反应15分钟。然后向上述反应体系中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(36.1毫克,0.077毫摩尔,1.2当量)和N,N-二异丙基乙胺(41.5毫克,0.32毫摩尔,5.0当量)。混合物在25摄氏度条件下继续反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,通过制备级高效液相色谱进行纯化,纯化条件:XBridge Shield RP18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.05%氨水),流动相B:乙腈;流速:25毫升/分钟;梯度:在8分钟内用56%→79%的B相梯度洗脱,检测器UV 254/220纳米;得到化合物25-7a(白色固体,60毫克,产率72%)。MS(ESI,m/z):1220.7/1222.7[M+H]
+。
步骤8
在零摄氏度搅拌条件下,向化合物25-7a(60.0毫克,0.047毫摩尔,1.0当量)的甲醇(2.0毫升)溶液中加入盐酸的1,4-二氧六环溶液(4摩尔/升,2.0毫升)。混合物在25摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.05%盐酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用5%→35%的B相梯度洗脱,检测器UV 254/220纳米;得到化合物25(淡黄色固体,40毫克,产率73%)。MS(ESI,m/z):1076.4/1078.5[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.61(s,1H),10.27–9.60(m,3H),9.06(s,1H),8.67–8.59(m,1H),8.01(d,J=1.6Hz,1H),7.94(d,J=9.3Hz,1H),7.81(d,J=8.3Hz,1H),7.47–7.35 (m,5H),7.31(d,J=2.4Hz,1H),7.26–7.18(m,2H),7.10(d,J=2.3Hz,1H),4.86–4.75(m,1H),4.74–4.64(m,1H),4.61–4.47(m,3H),4.48–4.30(m,3H),4.29–4.20(m,1H),4.20–4.14(m,2H),4.01–3.88(m,3H),3.67–3.53(m,5H),3.44(t,J=6.0Hz,3H),3.21–3.08(m,2H),2.44(s,3H),2.41–2.32(m,1H),2.30–2.20(m,1H),2.10–1.84(m,12H),1.01–0.85(m,10H);
19F NMR(377MHz,DMSO-d
6)δ-121.83。
实施例26
(2S,4S)-4-(4-((15-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-12-甲基-3,6,9-三氧-12-氮杂十五烷基)氧基)苯氧基)-1-((S)-2-环己基-2-((S)-2-(甲胺基)丙胺基)乙酰基)-N-((R)-1,2,3,4-四氢萘-1-基)吡咯烷-2-甲酰胺二甲酸盐26a;(2S,4S)-4-(4-((15-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-12-甲基-3,6,9-三氧-12-氮杂十五烷基)氧基)苯氧基)-1-((S)-2-环己基-2-((S)-2-(甲胺基)丙胺基)乙酰基)-N-((R)-1,2,3,4-四氢萘-1-基)吡咯烷-2-甲酰胺二甲酸盐26b。
步骤1
在25摄氏度搅拌条件下,向化合物L-环己基甘氨酸甲酯盐酸盐(10.00克,45.739毫摩尔,1.00当量)和N-叔丁氧羰基-N-甲基-丙氨酸(9.30克,45.739毫摩尔,1.00当量)的二氯甲烷(100毫升)溶液中加入3-(二乙氧基邻酰氧基)-1,2,3-苯并三嗪-4-酮(16.42克,54.887毫摩尔,1.20当量)和N,N-二异丙基乙胺(20.69克,160.087毫摩尔,3.50当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用30%→80%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV220纳米;得到化合物26-1(淡黄色油,9.0克,产率55%)。MS(ESI,m/z):357.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ6.65(s,1H),4.88–4.60(m,1H),4.54–4.40(m,1H),3.73(s,3H),2.79(s,3H),1.83–1.61(m,5H),1.58–1.52(m,1H),1.50(s,9H),1.33(d,J=7.1Hz,3H),1.28–1.13(m,2H),1.13–0.92(m,3H)。
步骤2
在25摄氏度搅拌条件下,向化合物26-1(9.00克,23.986毫摩尔,1.00当量)的四氢呋喃/甲醇/水(1/1/1,90毫升)溶液中加入氢氧化锂(906.96毫克,35.978毫摩尔,1.50当量)。所得混合物在25摄氏度下搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩除去四氢呋喃和甲醇。然后向所得粗产物中加入1摩尔每升的稀盐酸调节溶液pH至5,析出化合物26-2(白色固体,8.00克,77%)。MS(ESI,m/z):343.2[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ12.72(s,1H),7.95–7.39(m,1H),4.82–4.35(m,1H),4.27–4.03(m,1H),2.75(s,3H),1.81–1.50(m,6H),1.38(s,9H),1.30–0.89(m,8H)。
步骤3
在25摄氏度搅拌条件下,向化合物三缩四乙二醇(5.00克,25.743毫摩尔,1.00当量)的四氢呋喃(70毫升)溶液中加入咪唑(1.31克,19.307毫摩尔,0.75当量)和三异丙基氯硅烷(2.48克,12.872毫摩尔,0.50当量)。所得混合物在25摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→55%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物26-3(红色油,3克,产率33%)。MS(ESI,m/z):351.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ3.83(t,J=5.6Hz,2H),3.73–3.69(m,2H),3.69–3.62(m,8H),3.61–3.56(m,4H),1.14–1.00(m,21H)。
步骤4
在零摄氏度搅拌条件下,向(2S,4R)-1-(叔丁氧羰基)-4-羟基吡咯烷-2-羧酸(10.00克,41.081毫摩尔,1.00当量)的二氯甲烷(150毫升)溶液中加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(9.12克,45.190毫摩尔,1.10当量),1-羟基苯并三唑(7.01克,49.298毫摩尔,1.20当量),三乙胺(6.56克,61.622毫摩尔,1.50当量)和(R)-(-)-1,2,3,4-四氢-1-萘胺(7.00克,45.190毫摩尔,1.1当量)。所得混合物在25摄氏度下搅拌反应5小时。反应过程通过液质和薄层层析来监控。反应结束 后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物26-4(白色固体,14克,产率89%)。MS(ESI,m/z):361.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.22–6.99(m,4H),5.18–5.01(m,1H),4.54–4.32(m,2H),3.72–3.32(m,2H),2.96–2.69(m,3H),2.50–1.92(m,3H),1.90–1.64(m,3H),1.40(s,9H)。
步骤5
在25摄氏度,氮气保护搅拌条件下,向26-4(4.30克,11.333毫摩尔,1.00当量),4-苄氧基苯酚(3.11克,14.733毫摩尔,1.30当量),三丁基膦(3.62克,16.999毫摩尔,1.50当量)的甲苯(50毫升)溶液中加入偶氮二甲酰胺(3.08克,16.999毫摩尔,1.50当量)。所得混合物在60摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物26-5(白色固体,4克,产率65%)。MS(ESI,m/z):543.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.46–7.27(m,6H),7.19–7.12(m,1H),7.12–7.06(m,1H),7.05–6.97(m,1H),6.86–6.80(m,2H),6.61–6.56(m,2H),5.26–5.16(m,1H),5.01(s,2H),4.76–4.70(m,1H),4.46–4.41(m,1H),3.68–3.63(m,2H),2.87–2.71(m,2H),2.71–2.64(m,1H),2.12–2.01(m,1H),1.90–1.74(m,3H),1.67(s,1H),1.45(s,9H)。
步骤6
在25摄氏度搅拌条件下,向26-5(4.00克,7.002毫摩尔,1.00当量)的乙酸乙酯(40毫升)溶液中加入盐酸的1,4-二氧六环(4摩尔/升,40.00毫升)。所得混合物在25摄氏度下搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到化合物26-6(白色固体,3克,粗品)。粗产品直接用于下一步反应。MS(ESI,m/z):443.3[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向26-6(3.00克,6.440毫摩尔,1.00当量)和26-2(2.55克,7.084毫摩尔,1.10当量)的二氯甲烷(40毫升)溶液中加入3-(二乙氧基邻酰氧基)-1,2,3-苯并三嗪-4-酮(2.43克,7.728毫摩尔,1.20当量)和N,N-二异丙基乙胺(3.07克,22.539毫摩尔,3.50当量)。所得混合物在25摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→8%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物26-7(黄色固体,3克,产率60%)。MS(ESI,m/z):767.4[M+H]
+。
步骤8
25摄氏度,氮气保护搅拌条件下,向26-7(3.00克,3.716毫摩尔,1.00当量)的甲醇(40毫升)溶液中加入乙酸(1.17克,18.580毫摩尔,5.00当量)和氢氧化钯/碳(20%钯含量,500毫克)。通过置换气将氮气置换成氢气。所得混合物在氢气氛围(1.5大气压)中于25摄氏度下搅拌反应4小时。反应过程通过液质和薄层层析来监控。反应结束后,过滤除去不溶物,滤饼用甲醇(20毫升)清洗,滤液减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→95%的乙腈/水流动相(0.1%甲酸)进行洗脱;检测器UV220纳米;得到化合物26-8(白色固体,2.0毫克,产率79%)。MS(ESI,m/z):677.5[M+H]
+。
步骤9
在25摄氏度,氮气保护搅拌条件下,向26-8(500.0毫克,0.702毫摩尔,1.00当量),26-3(336.66毫克,0.912毫摩尔,1.30当量),三丁基膦(298.91毫克,1.404毫摩尔,2.00当量)的甲苯(10毫升)溶液中加入偶氮二甲酰胺(254.39毫克,1.404毫摩尔,2.00当量)。所得混合物在60摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶 剂,得到化合物26-9(白色固体,400毫克,产率56%)。MS(ESI,m/z):1009.8[M+H]
+。
步骤10
在25摄氏度搅拌条件下,向化合物26-9(400.00毫克,0.376毫摩尔,1.00当量)的四氢呋喃(4.0毫升)溶液中加入四丁基氟化铵(124.33毫克,0.452毫摩尔,1.20当量)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物26-10(无色油,300毫克,产率93%)。MS(ESI,m/z):853.6[M+H]
+。
步骤11
在25摄氏度搅拌条件下,向化合物26-10(300.00毫克,0.334毫摩尔,1.00当量)的乙腈(6.0毫升)溶液中加入2-碘酰基苯甲酸(147.71毫克,0.501毫摩尔,1.50当量)。混合物在60摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至室温。反应液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物26-11(白色固体,250毫克,产率87%)。MS(ESI,m/z):851.5[M+H]
+。
步骤12
在零摄氏度搅拌条件下,向化合物26-11(70.00毫克,0.078毫摩尔,1.00当量)和2-9a(39.22毫克,0.063毫摩尔,0.80当量)的甲醇(1.5毫升)溶液中加入乙酸(4.94毫克,0.078毫摩尔,1.00当量)和氰基硼氢化钠(10.34毫克,0.156毫摩尔,2.00当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到中间体。在25摄氏度搅拌条件下,向该中间体中加入二氯甲烷(1.5毫升)和三氟乙酸(0.5毫升)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 150毫 米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用85%→95%B相梯度洗脱;检测器UV 254/220纳米;得到28a(白色固体,14.1毫克,产率13%)。MS(ESI,m/z):1230.7/1232.7[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.51(s,2H),7.96(d,J=1.6Hz,1H),7.75(d,J=8.4Hz,1H),7.50–7.39(m,1H),7.29–7.16(m,4H),7.14–7.03(m,4H),6.82–6.76(m,4H),5.01–4.92(m,3H),4.59–4.52(m,3H),4.46–4.41(m,1H),4.24–4.20(m,1H),4.02–3.97(m,6H),3.82–3.74(m,5H),3.66–3.52(m,9H),3.28–3.19(m,7H),2.81(s,3H),2.74–2.67(m,2H),2.54(s,1H),2.49–2.34(m,3H),2.38–2.33(m,1H),2.22–2.19(m,2H),1.93–1.59(m,10H),1.44–1.34(m,3H),1.20–0.99(m,5H)。
步骤12’
在零摄氏度搅拌条件下,向化合物26-11(50.00毫克,0.056毫摩尔,1.00当量)和2-9b(28.02毫克,0.045毫摩尔,0.80当量)的甲醇(1.0毫升)溶液中加入乙酸(3.35毫克,0.056毫摩尔,1.00当量)和氰基硼氢化钠(7.38毫克,0.112毫摩尔,2.00当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到中间体。在25摄氏度搅拌条件下,向该中间体中加入二氯甲烷(1.0毫升)和三氟乙酸(0.3毫升)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.1%三氟乙酸),流动相B:乙腈;流速:25毫升/分钟;梯度:在7分钟内用85%→95%B相梯度洗脱;检测器UV 254/220纳米;得到28b(白色固体,5.8毫克,产率8%)。MS(ESI,m/z):1230.7/1232.7[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.37(s,2H),7.88(d,J=1.6Hz,1H),7.66(d,J=8.4Hz,1H),7.34–7.29(m,1H),7.19–7.07(m,4H),7.05–6.94(m,4H),6.74–6.63(m,4H),4.92–4.82(m,3H),4.49–4.44(m,3H),4.38–4.32(m,1H),4.14–4.10(m,1H),3.94–3.89(m,6H),3.81–3.59(m,6H),3.58–3.51(m,8H),3.25–3.22(m,7H),2.77(s,3H),2.69–2.59(m,2H),2.50–2.44(m,3H),2.39–2.24(m,2H),2.17–2.10(m,2H),1.87–1.77(m,1H),1.72–1.57(m,7H),1.52–1.50(m,2H),1.39–1.29(m,3H),1.19–0.87(m,5H)。
实施例27
(2R,3S,4R,5S)-3-(3-氯-2-氟苯基)-4-(4-氯-2-氟苯基)-N-(4-((15-((S或R)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基)喹唑啉-2-基)氧基)-12-甲基-3,6,9-三氧基-12-氮杂十五烷基)氨甲酰基)-2-甲氧基苯基)-4-氰基-5-新戊基吡咯烷-2-甲酰胺27a;(2R,3S,4R,5S)-3-(3-氯-2-氟苯基)-4-(4-氯-2-氟苯基)-N-(4-((15-((R或S)-6-氯-8-氟-7-(3-羟基萘-1-基)-4-(哌嗪-1-基) 喹唑啉-2-基)氧基)-12-甲基-3,6,9-三氧基-12-氮杂十五烷基)氨甲酰基)-2-甲氧基苯基)-4-氰基-5-新戊基吡咯烷-2-甲酰胺27b。
步骤1
在25摄氏度搅拌条件下,向50毫升圆底烧瓶中依次加入4-((2R,3S,4R,5S)-3-(3-氯-2-氟苯基)-4-(4-氯-2-氟苯基)-4-氰基-5-新戊基吡咯烷-2-甲酰胺基)-3-甲氧基苯甲酸(400.0毫克,0.6毫摩尔,1.0当量),二氯甲烷(10.0毫升),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(296.1毫克,0.7毫摩尔,1.2当量),1-氨基-3,6,9-三噁-11-十一醇(150.5毫克,0.7毫摩尔,1.2当量)和N,N-二异丙基乙胺(251.6克,1.9毫摩尔,3.0当量)。混合物在25摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应完成后,减压浓缩得到粗产品,粗产品通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→60%的乙腈/水(0.1%碳酸氢铵)流动相进行洗脱;检测器UV254纳米;得到化合物27-1(白色固体,490.0毫克,产率90%)。MS(ESI,m/z):791.2/793.2[M+H]
+;
1HNMR(400MHz,DMSO-d
6)δ10.42(s,1H),8.52(t,J=5.6Hz,1H),8.32(d,J=8.4Hz,1H),7.78–7.71(m,1H),7.62–7.47(m,4H),7.44–7.32(m,3H),4.67–4.56(m,3H),4.43–4.34(m,1H),4.00–3.90(m,4H),3.67–3.48(m,11H),3.42–3.38(m,4H),1.68–1.62(m,1H),1.27(d,J=13.9Hz,1H),0.98(s,9H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向50毫升反应瓶中依次加入化合物27-1(300毫克,0.4毫摩尔,1.0当量),乙腈(10.0毫升)和2-碘酰基苯甲酸(159.2毫克,0.5毫摩尔,1.5当量)。所得混合物在60摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩除去多余的试剂得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物27-2(白色固体,290.0毫克,产率92%)。MS(ESI,m/z):789.2/791.2[M+H]
+。
步骤3
在25摄氏度搅拌条件下,向化合物2-9a(30.0毫克,0.05毫摩尔,1.0当量)的甲醇(5.0毫升)溶液中加入27-2(48.0毫克,0.06毫摩尔,1.3当量),乙酸(4.2毫克,0.07毫摩尔,1.5当量)和氰基硼氢化钠(8.8毫克,0.14毫摩尔,3.0当量)。混合物在25摄氏度下反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩除得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物27-3a(白色固体,52.0毫克,产率86%)。MS(ESI,m/z):1368.4/1370.4[M+H]
+。
步骤4
在25摄氏度搅拌条件下,向化合物27-3a(52.0毫克,0.04毫摩尔,1.0当量)的二氯甲烷(2.0 毫升)溶液中加入三氟乙酸(2.0毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:色谱柱XSelect CSH Prep C18 OBD,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;在7分钟内用20%→50%的B相进行洗脱,检测器UV220/235纳米;得到化合物27a(白色固体,28.4毫克,产率59%)。MS(ESI,m/z):1268.6/1270.6[M+H]
+;
1HNMR(400MHz,DMSO-d
6)δ10.41(s,1H),8.49–8.46(m,1H),8.32(d,J=8.4Hz,1H),8.20(s,1H),7.95(d,J=1.5Hz,1H),7.81(d,J=8.3Hz,1H),7.76–7.70(m,1H),7.61–7.51(m,3H),7.50–7.47(m,1H),7.46–7.41(m,1H),7.39–7.33(m,3H),7.29(d,J=2.4Hz,1H),7.24–7.18(m,2H),7.06(d,J=2.3Hz,1H),4.62–4.55(m,2H),4.41–4.32(m,3H),3.98–3.93(m,1H),3.91(s,3H),3.86–3.79(m,4H),3.50–3.42(m,12H),3.41–3.36(m,2H),3.04–2.98(m,4H),2.49–2.46(m,2H),2.19(s,3H),1.92–1.83(m,2H),1.68–1.60(m,1H),1.34–1.20(m,2H),0.97(s,9H)。
步骤3’
在25摄氏度搅拌条件下,向化合物2-9b(40.0毫克,0.06毫摩尔,1.0当量)的甲醇(5.0毫升)溶液中加入27-2(64.0毫克,0.08毫摩尔,1.3当量),乙酸(5.6毫克,0.09毫摩尔,1.5当量)和氰基硼氢化钠(11.7毫克,0.2毫摩尔,3.0当量)。混合物在25摄氏度下反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物27-3b(白色固体,52.0毫克,产率61%)。MS(ESI,m/z):1368.4/1370.4[M+H]
+。
步骤4’
在25摄氏度搅拌条件下,向化合物27-3b(52.0毫克,0.04毫摩尔,1当量)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(2.0毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,混合物通过制备级高效液相色谱进行纯化,纯化条件:色谱柱XSelect CSH Prep C18 OBD,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈; 流速:25毫升/分钟;在7分钟内用20%→50%的B相进行洗脱,检测器UV 220/235纳米;得到化合物27b(白色固体,19.1毫克,产率40%)。MS(ESI,m/z):1268.6/1270.6[M+H]
+;
1HNMR(400MHz,DMSO-d
6)δ10.41(s,1H),8.52–8.44(m,1H),8.32(d,J=8.4Hz,1H),8.19(s,1H),7.96(d,J=1.6Hz,1H),7.84–7.69(m,2H),7.62–7.32(m,8H),7.29(d,J=2.4Hz,1H),7.26–7.17(m,2H),7.07(d,J=2.3Hz,1H),4.62–4.55(m,2H),4.42–4.31(m,3H),4.01–3.94(m,1H),3.91(s,3H),3.87–3.80(m,4H),3.52–3.44(m,12H),3.43–3.36(m,2H),3.07–3.00(m,4H),2.49–2.46(m,2H),2.20(s,3H),1.93–1.81(m,2H),1.70–1.58(m,1H),1.32–1.22(m,2H),0.97(s,9H)。
实施例28
(2S,4R)-1-((S)-2-(叔丁基)-17-(4-((S或R)-6-氯-2-(3-(二甲胺基)氮杂环丁烷-1-基)-8-氟-7-(3-羟基萘-1-基)喹唑啉-4-基)哌嗪-1-基)-4-氧代-6,9,12,15-四氧代-3-氮杂庚烷酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺28a;(2S,4R)-1-((S)-2-(叔丁基)-17-(4-((R或S)-6-氯-2-(3-(二甲胺基)氮杂环丁烷-1-基)-8-氟-7-(3-羟基萘-1-基)喹唑啉-4-基)哌嗪-1-基)-4-氧代-6,9,12,15-四氧代-3-氮杂庚烷酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺28b。
步骤1
在25摄氏度搅拌条件下,向化合物乙酸叔丁酯-四聚乙二醇(1.0克,3.2毫摩尔,1.0当量)的二氯甲烷(10.0毫升)溶液中加入三氟乙酸(3.0毫升)。混合物在25摄氏度下搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品28-1(明亮的黄色油状物,820.0毫克)。粗产品直接用于下一步反应。MS(ESI,m/z):253.1[M+H]
+。
步骤2
在25摄氏度搅拌条件下,向化合物28-1(818.5毫克,1.0当量)的N,N-二甲基甲酰胺(8.0毫 升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(1.4克,3.6毫摩尔,1.1当量)。混合物在25摄氏度条件下反应30分钟。然后向反应液中加入N,N-二异丙基乙胺(2.1克,16.3毫摩尔,5.0当量)和(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(1.3克,2.6毫摩尔,0.8当量)。混合物在25摄氏度条件下反应3小时,反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩除去多余的试剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢氨)进行洗脱;检测器UV254/220纳米;得到化合物28-2(亮黄色固体,948.5毫克,产率40%)。MS(ESI,m/z):679.4[M+H]
+。
步骤3
在25摄氏度,氮气保护搅拌条件下,向100毫升反应瓶中依次加入化合物28-2(785.0毫克,1.2毫摩尔,1.0当量),乙腈(8.0毫升)和2-碘酰基苯甲酸(356.4毫克,1.3毫摩尔,1.5当量)。所得混合物在60摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物28-3(白色固体,615.3毫克,产率79%)。MS(ESI,m/z):677.2[M+H]
+。
步骤4
在25摄氏度,氮气保护搅拌条件下,向化合物1-7(500.0毫克,1.0毫摩尔,1.0当量)的N-甲基吡咯烷酮(5.0毫升)溶液中加入3-(二甲氨基)氮杂环丁烷二盐酸盐(135.6毫克,1.3毫摩尔,1.3当量),N,N-二异丙基乙胺(722.4毫克,5.6毫摩尔,4.0当量)。混合物在60摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至室温。反应液直接通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的乙腈/水(0.1%碳酸氢铵)流动相进行洗脱;检测器UV254/220纳米;得到化合物28-4(白色固体,900毫克,产率29%)。MS(ESI,m/z):543.1/545.0/547.0[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.62–7.62(m,1H),4.36–4.16(m,2H), 4.15–3.96(m,2H),3.75–3.50(m,8H),3.28–3.14(m,1H),2.24(s,6H),1.49(s,9H)。
步骤5
在25摄氏度,氮气保护搅拌条件下,向化合物28-4(900.0毫克,1.7毫摩尔,1.0当量)的四氢呋喃/水(10/1,11.0毫升)溶液中加入4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(535.0毫克,2.0毫摩尔,1.2当量),碳酸钾(684.7毫克,5.0毫摩尔,3.0当量)和氯(2-二环己基膦基-2',4',6'-三异丙基-1,1'-联苯基)[2-(2'-氨基-1,1'-联苯)]钯(II)(129.7毫克,0.2毫摩尔,0.1当量)。反应液在60摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液冷却至室温。反应液减压浓缩得到粗产品,粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物28-5(二个立体异构体混合物,白色固体,600.0毫克,产率60%)。MS(ESI,m/z):607.2/609.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.70–7.62(m,2H),7.60–7.55(m,1H),7.38–7.27(m,2H),7.23–7.20(m,1H),7.19–7.08(m,1H),4.20–4.40(m,2H),4.16–3.98(m,2H),3.82–3.50(m,8H),3.22–3.08(m,1H),2.18(s,6H),1.530(s,9H)。
步骤6
通过制备级手性高效液相色谱法对步骤5所得化合物28-5(600.0毫克)进行手性拆分:手性柱CHIRALPAK IC,2 x 25厘米,5微米;流动相A:正己烷/甲基叔丁基醚=1/1(0.5%2摩尔/升氨甲醇),流动相B:乙醇;流速:20毫升/分钟;在11.4分钟内用10%的B相进行洗脱,检测器UV 225/254纳米。得到两个产品,较短保留时间(6.8分钟)的产品为化合物28-5a(白色固体,225.0毫克,回收率37%),化合物28-5a:MS(ESI,m/z):607.2/609.2[M+H]
+;较长保留时间(9分钟)的产品为化合物28-5b(白色固体,230.0毫克,回收率38%),化合物28-5b:MS(ESI,m/z):607.2/609.2[M+H]
+。
步骤7
在零摄氏度搅拌条件下,向化合物28-5a(225.0毫克,0.4毫摩尔,1.0当量)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)。混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品用高效液相色谱纯化,纯化条件:色谱柱Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米,流动相A:水(10摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;用10%的流动相B洗脱2分钟,然后用10%→26%流动相B洗脱2.5分钟,最后再用26%→55%流动相B洗脱2分钟。检测器UV220纳米;得到产品28-6a(白色固体,150.0毫克,产率80%)。MS(ESI,m/z):507.2/509.1[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ10.00(s,1H),7.83–7.72(m,2H),7.49–7.38(m,1H),7.30–7.18(m,3H),7.04(d,J=2.4Hz,1H),4.16–4.05(m,2H),3.92–3.81(m,2H),3.70–3.60(m,4H),3.18–3.06(m,1H),2.96–2.82(m,4H),2.11(s,6H)。
步骤7’
在零摄氏度搅拌条件下,向化合物28-5b(230.0毫克,0.4毫摩尔,1.0当量)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)。混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过高效液相色谱纯化,纯化条件:色谱柱Gemini-NX C18 AXAI Packed,21.2 x 150毫米,5微米,流动相A:水(10摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;用10%流动相B洗脱2分钟,然后用10%→26%的流动相B洗脱2.5分钟,最后再用26%→55%流动相B洗脱2分钟。检测器UV220纳米;得到产品28-6b(白色固体,145.0毫克,产率76%)。MS(ESI,m/z):507.2/509.1[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ9.99(s,1H),7.84–7.72(m,2H),7.49–7.38(m,1H),7.30–7.17(m,3H),7.04(d,J=2.4Hz,1H),4.17–4.04(m,2H),3.93–3.81(m,2H),3.76–3.63(m,4H),3.18–3.07(m,1H),3.03–2.89(m,4H),2.12(s,6H)。
步骤8
在25摄氏度搅拌条件下,向化合物28-6a(40.0毫克,0.08毫摩尔,1.0当量)的甲醇(2.0毫升)溶液中加入氰基硼氢化钠(7.8毫克,0.1毫摩尔,1.5当量)和乙酸(7.5毫克,0.1毫摩尔,1.5当量)。然后将化合物28-3(64.8毫克,0.1毫摩尔,1.2当量)溶于甲醇(2.0毫升)中,缓慢滴加至反应液中。混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,9 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用32%→61%的流动相B洗脱7分钟;检测器UV254/220纳米;得到产品28a(白色固体,31.0毫克,产率34%)。MS(ESI,m/z):1167.4/1169.4[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.90–8.86(m,1H),7.82–7.72(m,2H),7.48–7.34(m,5H),7.31–7.13(m,3H),7.04(d,J=2.4Hz,1H),5.07–4.97(m,1H),4.70(s,1H),4.63–4.53(m,1H),4.48–4.41(m,1H),4.32–4.22(m,2H),4.09–3.99(m,4H),3.89–3.80(m,5H),3.79–3.63(m,15H),3.30–3.20(m,1H),2.86–2.65(m,6H),2.48(s,3H),2.26(s,6H),2.24–2.16(m,1H),2.10–1.90(m,1H),1.56–1.48(m,3H),1.11–1.01(m,9H)。
步骤8’
在25摄氏度搅拌条件下,向化合物28-6b(50.0毫克,0.1毫摩尔,1.0当量)的甲醇(2.0毫升)溶液中加入氰基硼氢化钠(9.3毫克,0.1毫摩尔,1.5当量)和乙酸(8.9毫克,0.2毫摩尔,1.5当量)。然后将化合物28-3(80.1毫克,0.1毫摩尔,1.2当量)溶于甲醇(2.0毫升),缓慢滴加至反应液中。混合物在25摄氏度条件下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过制备级高效液相色谱进行纯化,纯化条件:XSelect CSH Prep C18 OBD Column,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;用32%→61%的流动相B洗脱7分钟;检测器UV254/220纳米;得到产品28b(白色固体,31.0毫克,产率28%)。MS(ESI,m/z):1167.4/1169.4[M+H]
+;
1H NMR(300MHz,CD
3OD)δ8.90–8.86(m,1H),7.82–7.72(m,2H),7.48–7.34(m,5H),7.31–7.14(m,3H),7.04(d,J=2.4Hz,1H),5.08–4.96(m,1H),4.70(s,1H),4.63–4.53(m,1H),4.48–4.41(m,1H),4.32–4.22(m,2H),4.09–3.99(m,4H),3.90– 3.80(m,5H),3.79–3.62(m,15H),3.30–3.20(m,1H),2.86–2.65(m,6H),2.48(s,3H),2.26(s,6H),2.24–2.16(m,1H),2.10–1.90(m,1H),1.56–1.48(m,3H),1.11–1.01(m,9H)。
实施例29
(2S,4R)-1-((S)-2-(2-(2-((S)-2-((((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐29
化合物29参照实施例25合成得到。化合物29(淡黄色固体)。MS(ESI,m/z):1048.5/1050.5[M+H]
+。
1H NMR(400MHz,DMSO-d
6)δ10.86–9.78(m,3H),9.09(s,1H),8.70–8.61(m,1H),8.01(d,J=1.7Hz,1H),7.81(d,J=8.4Hz,1H),7.73(d,J=9.3Hz,1H),7.48–7.36(m,5H),7.32(d,J=2.4Hz,1H),7.27–7.17(m,2H),7.12(d,J=2.4Hz,1H),4.86–4.66(m,2H),4.59–4.32(m,6H),4.29–3.82(m,10H),3.80–3.56(m,4H),3.49–3.38(m,1H),3.31–3.20(m,1H),2.45(s,3H),2.31–2.16(m,1H),2.11–1.81(m,9H),0.92(s,9H);
19F NMR(377MHz,DMSO-d
6)δ-121.87。
实施例30
(2S,4R)-1-((S)-2-(3-((S)-2-(((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐30
化合物30参照实施例25合成得到。化合物30(淡黄色固体)。MS(ESI,m/z):1018.6/1020.5[M+H]
+。
1H NMR(300MHz,DMSO-d
6)δ10.74(s,1H),9.99(s,1H),9.74(s,1H),9.05(s,1H),8.68–8.58 (m,1H),8.27(d,J=9.2Hz,1H),8.01(d,J=1.6Hz,1H),7.82(d,J=8.3Hz,1H),7.49–7.28(m,6H),7.27–7.15(m,2H),7.11(d,J=2.4Hz,1H),4.88–4.64(m,3H),4.60–4.50(m,4H),4.27–4.23(m,1H),4.21–4.09(m,3H),4.07–3.88(m,3H),3.76–3.47(m,4H),3.44–3.28(m,1H),3.24–3.09(m,1H),2.95–2.73(m,2H),2.45(s,3H),2.34–2.15(m,1H),2.12–1.79(m,9H),0.91(s,9H);
19F NMR(282MHz,DMSO-d
6)δ-121.79。
实施例31
(2S,4R)-1-((S)-2-(4-((S)-2-((((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丁酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐31
化合物31参照实施例25合成得到。化合物31(类白色固体)。MS(ESI,m/z):1032.6/1034.6[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ10.71(s,1H),10.09–9.99(m,1H),9.84–9.69(m,1H),9.12(s,1H),8.71–8.61(m,1H),8.16–8.05(m,2H),7.88(d,J=8.3Hz,1H),7.55–7.42(m,5H),7.38(d,J=2.4Hz,1H),7.34–7.24(m,2H),7.17(d,J=2.4Hz,1H),4.90–4.72(m,2H),4.68–4.36(m,6H),4.32(d,J=5.6Hz,1H),4.27–4.21(m,2H),4.09–3.95(m,3H),3.77–3.61(m,3H),3.55–3.40(m,1H),3.28–3.08(m,2H),2.52(s,3H),2.46–2.26(m,3H),2.01(d,J=11.3Hz,11H),0.98(s,9H);
19F NMR(282MHz,DMSO-d
6)δ-121.80。
实施例32
(2S,4R)-1-((2S)-2-(4-(4-((2S)-2-((((7S或7R)-4-(3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丁氧基)丁酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐32
化合物32参照实施例25合成得到。化合物32(淡黄色固体)。MS(ESI,m/z):1104.5/1106.5[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.47–10.34(m,1H),9.95–9.82(m,1H),9.66–9.59(m,1H),9.02(s,1H),8.64–8.54(m,1H),8.01(d,J=1.6Hz,1H),7.94–7.76(m,2H),7.49–7.35(m,5H),7.31(d,J=2.4Hz,1H),7.26–7.14(m,2H),7.09(d,J=2.4Hz,1H),4.80–4.65(m,2H),4.59–4.49(m,3H),4.47–4.38(m,2H),4.37–4.31(m,1H),4.26–4.14(m,3H),4.00–3.85(m,4H),3.52–3.38(m,3H),3.34–3.26(m,4H),3.17–3.06(m,2H),2.44(s,3H),2.29–2.13(m,3H),2.08–1.86(m,9H),1.82–1.63(m,4H),1.57–1.46(m,2H),0.92(s,9H);
19F NMR(377MHz,DMSO-d
6)δ-121.88。
实施例33
(2S,4R)-1-((S)-2-(7-((S)-2-((((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)庚酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐33
化合物33参照实施例25合成得到。化合物33(淡黄色固体)。MS(ESI,m/z):1174.6/1176.6[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ9.01(s,1H),8.00(s,1H),7.80(d,J=8.3Hz,1H),7.49–7.33(m,5H),7.31(d,J=2.4Hz,1H),7.27–7.14(m,2H),7.06(d,J=2.4Hz,1H),4.75–4.62(m,2H),4.60–4.47(m,3H),4.45–4.37(m,2H),4.37–4.31(m,1H),4.26–4.15(m,3H),3.93–3.87(m,3H),3.66–3.62(m,2H),3.45–3.34(m,1H),3.21–3.00(m,2H),2.42(s,3H),2.31–2.16(m,2H),2.14–1.79(m,10H),1.71–1.59(m,2H),1.51–1.36(m,2H),1.35–1.14(m,5H),0.89(s,9H);
19F NMR(282MHz,DMSO-d
6)δ-121.87。
实施例34
(2S,4R)-1-((2S)-2-(2-(5-(2-((2S)-2-(((7S或7R)-4-(3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙基)四氢呋喃-2-基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二甲酸盐34;(2S,4R)-1-((2S)-2-(2-((2S或R,5S或R)-5-(2-((2S)-2-(((7S或7R)-4-(3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙基)四氢呋喃-2-基)乙酰胺-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺甲酸盐34aa;(2S,4R)-1-((2S)-2-(2-((2S或R,5R或S)-5-(2-((2S)-2-(((7S或7R)-4-(3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯 烷-1-基)乙基)四氢呋喃-2-基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺甲酸盐34ab;(2S,4R)-1-((2S)-2-(2-((2R或S,5S或R)-5-(2-((2S)-2-(((7S或7R)-4-(3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙基)四氢呋喃-2-基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二甲酸盐34ba;(2S,4R)-1-((2S)-2-(2-((2R或S,5R或S)-5-(2-((2S)-2-(((7S或7R)-4-(3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙基)四氢呋喃-2-基)乙酰胺-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺甲酸盐34bb
步骤1:
在零摄氏度搅拌条件下,向2-碘乙酸乙酯(20.0克,88.8毫摩尔,1.0当量),呋喃(114.5克,1598.1毫摩尔,18.0当量)和七水合硫酸亚铁(13.0克,44.4毫摩尔,0.5当量)的二甲基亚砜(200 毫升)溶液中加入双氧水(30%,22.2克,195.3毫摩尔,2.2当量)。混合物在20摄氏度搅拌条件下反应4小时,反应过程通过液质和薄层层析来监控。反应结束后,向反应液中加入水(1200毫升),再用乙酸乙酯(800毫升 x 3)萃取,合并有机相;有机相用饱和食盐水(800毫升 x 3)清洗,再用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→30%甲基叔丁基醚/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂得到化合物34-1(无色油状物,10克,产率69%)。MS(ESI,m/z):155.0[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.40–7.32(m,1H),6.35–6.32(m,1H),6.24–6.21(m,1H),4.23–4.15(m,2H),3.68(d,J=2.1Hz,2H),1.31–1.25(m,3H)。
步骤2:
在零摄氏度氮气保护搅拌条件下,向化合物34-1(10.0克,61.6毫摩尔,1.0当量)的无水四氢呋喃(100毫升)溶液中缓慢滴加氢化铝锂的四氢呋喃溶液(2.5摩尔/升,49毫升,123.2毫摩尔,2.0当量)。混合物在0摄氏度条件下反应1.5小时,反应过程通过液质和薄层层析来监控。反应结束后,在0摄氏度条件下向反应液中依次缓慢滴加水(4.9克),10%的氢氧化钠水溶液(4.9克)和水(14.7克)。过滤除去不溶物,滤饼用四氢呋喃(60毫升 x 3)清洗,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→30%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂得到化合物34-2(黄色油状物,4.86克,产率66%)。
1H NMR(400MHz,CDCl
3)δ7.39–7.29(m,1H),6.35–6.30(m,1H),6.14–6.06(m,1H),3.88(t,J=6.2Hz,2H),2.90(t,J=6.2Hz,2H)。
步骤3:
在零摄氏度搅拌条件下,向化合物34-2(4.45克,37.7毫摩尔,1.0当量),2-碘乙酸乙酯(8.5克,37.7毫摩尔,1.0当量)和七水合硫酸亚铁(5.5克,18.9毫摩尔,0.5当量)的二甲基亚砜(80毫升)溶液中缓慢加入双氧水(30%,9.4克,83.0毫摩尔,2.2当量)。混合物在20摄氏度条件下反应16小时,反应过程通过液质和薄层层析来监控。反应结束后,向反应液中加入水(500毫升),再用乙酸乙酯(400毫升 x 3)萃取,合并有机相;有机相用饱和食盐水(800毫升 x 3)清洗,再用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→50%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物34-3(黄色油状物,3.8克,产率45%)。MS(ESI,m/z):199.0[M+H]
+;
1H NMR(400MHz,CDCl
3)δ6.13(d,J=3.1Hz,1H),6.05(d,J=3.1Hz,1H),4.22–4.15(m,2H),3.87–3.81(m,2H),3.64(s,2H),2.90–2.80(m,2H),1.84(s,1H),1.30–1.23(m,3H)。
步骤4:
在25摄氏度氮气保护搅拌条件下,向化合物34-3(3.8克,18.2毫摩尔,1.0当量)的乙醇(50毫升)溶液中加入铑/三氧化二铝(5%,1.9克,0.9毫摩尔,0.05当量)。然后将氮气置换成氢气。混合物在25摄氏度,5个大气压的氢气环境中反应40小时,反应过程通过液质和薄层层析来监控。反应结束后,过滤除去不溶物,滤饼用乙醇(20 x 3毫升)清洗,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,用0%→70%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物34-4(黄色油状物,2.05克,产率52%)。MS(ESI,m/z):203.0[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.33–4.21(m,1H),4.20–4.11(m,2H),4.11–4.02(m,1H),3.81–3.75(m,2H),2.66–2.55(m,1H),2.54–2.43(m,2H),2.16–1.98(m,2H),1.90–1.73(m,2H),1.72–1.55(m,2H),1.27(t,J=7.1Hz,3H)。
步骤5:
在零摄氏度搅拌条件下,向化合物34-4(2.0克,9.4毫摩尔,1.0当量),4-二甲氨基吡啶(0.12克,0.94毫摩尔,0.1当量)和三乙胺(2.0克,18.8毫摩尔,2.0当量)的二氯甲烷(20毫升)溶液中缓慢加入对甲苯磺酰氯(2.1克,10.3毫摩尔,1.1当量)。反应液在25摄氏度条件下反应16小时,反应过程通过液质和薄层层析来监控。反应结束后,向体系中加入水(20毫升)淬灭反应,水相用二氯甲烷(20毫升 x 3)萃取,合并有机相;有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→50%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物34-5(无色油状物,3.2克,产率90%)。MS(ESI,m/z):357.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.83–7.76(m,2H),7.40–7.32(m,2H),4.21–4.07(m,5H),3.94–3.80(m,1H),2.58–2.34(m,5H),2.12–1.80(m,4H),1.63–1.46(m,2H),1.30–1.22(m,3H)。
步骤6:
在25摄氏度搅拌条件下,将化合物34-5(3.0克,8.0毫摩尔,1.0当量)溶于30毫升的乙腈中,然后加入L-脯氨醇(1.02克,9.6毫摩尔,1.2当量)和碳酸钾(2.33克,16毫摩尔,2.0当量)。混合物在60摄氏度下反应4小时,反应过程通过液质和薄层层析来监控。反应结束后,过滤除去不溶物,滤饼用乙腈(15 x 2毫升)清洗,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→10%氨甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物34-6(黄色油状物,2.27克,产率94%)。MS(ESI,m/z):286.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ4.29–4.19(m,1H),4.19–4.08(m,2H),4.02–3.85(m,1H),3.68–3.59(m,1H),3.45–3.33(m,1H),3.27–3.14(m,1H),2.95–2.80(m,1H),2.66–2.56(m,2H),2.49–2.21(m,3H),2.15–1.97(m,2H),1.91–1.49(m,9H),1.29–1.24(m,3H)。
步骤7:
在25摄氏度搅拌条件下,将化合物34-6(1.2克,4.0毫摩尔,1.0当量)溶于四氢呋喃/甲醇/水的混合溶液中(1/1/1,12毫升),然后加入氢氧化锂(0.2克,7.9毫摩尔,2.0当量)。混合物在25摄氏度下反应16小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩除去溶剂,然后加入12毫升的水稀释混合物。在0摄氏度下,滴加稀盐酸(1摩尔/升,8毫升)调节pH至5,然后减压浓缩得到粗产品化合物34-7(1.4克)。所得粗产品未经纯化,直接用于下一步反应。MS(ESI,m/z):258.1[M+H]
+。
步骤8:
在零摄氏度氮气保护条件下,向化合物16-3a(1.0克,1.5毫摩尔,1.0当量)和化合物34-7(0.71克,1.8毫摩尔,1.2当量,纯度67%)的无水四氢呋喃(10毫升)溶液中加入叔丁醇钾的四氢呋喃溶液(1摩尔/升,3.9毫升,3.9毫摩尔,2.5当量)。混合物在零摄氏度条件下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,粗产品通过反相色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器,UV254/220纳米;得到化合物34-8(黄色固体,700毫克,产率51%)。MS(ESI,m/z):834.4/836.4[M+H]
+。
步骤9:
在25摄氏度搅拌条件下,向化合物34-8(700毫克,0.84毫摩尔,1.0当量)的N,N-二甲基甲酰胺(8毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(383毫克,1.01毫摩尔,1.2当量)。混合物在25摄氏度下反应15分钟。然后向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(472毫克,1.01毫摩尔,1.2当量)和N,N-二异丙基乙胺(433毫克,3.36毫摩尔,4当量)。反应液在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,向反应瓶中加入水(60毫升),然后用乙酸乙酯(60毫升 x 3)萃取。合并有机相,有机相用饱和食盐水清洗(30毫升 x 3),有机相用无水硫酸钠干燥,过滤除去干燥剂;滤液减压浓缩得到粗产品。混合物通过反相色谱柱(C18柱)进行纯化,在25分钟内用40%→75%的甲醇/水流动相(0.1%碳酸氢铵)进行洗脱;检测器,UV254/220纳米;得到化合物34-9(黄色固体,750毫克,产率72%)。MS(ESI,m/z):1246.5/1248.5[M+H]
+。
步骤10:
在0摄氏度搅拌条件下,向化合物34-9(50毫克,0.038毫摩尔,1.0当量)的甲醇(2毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,2毫升)。混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品。粗产品溶于甲醇(2毫升),并在0摄氏度搅拌条件下加入氨甲醇溶液(7摩尔/升,2毫升)。混合物在25摄氏度条件下搅拌20分钟后,然后减压浓缩。粗产品溶于甲醇(2毫升),并在0摄氏度搅拌条件下加入甲酸(2毫升)。混合物在25摄氏度条件下搅拌10分钟后,然后减压浓缩得到粗产品。所得粗产品通过高效液相色谱进行纯化,色谱柱Sunfire prep C18 column,30 x 150毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:60毫升/分钟;在7分钟内用32%→51%的B相进行洗脱,检测器UV 220/254纳米。得到化合物34(白色固体,14毫克,产率30%)。MS(ESI,m/z):1102.1/1104.1[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ8.97–8.94(m,1H),8.63–8.57(m,1H),8.29(s,2H),7.98–7.75(m,3H), 7.48–7.34(m,5H),7.30(d,J=2.4Hz,1H),7.27–7.17(m,2H),7.07(d,J=2.4Hz,1H),4.58–3.96(m,11H),3.81–3.63(m,7H),3.09–2.81(m,3H),2.48–2.34(m,5H),2.28–2.16(m,2H),2.08–1.99(m,1H),1.96–1.82(m,4H),1.81–1.58(m,9H),1.53–1.38(m,2H),0.96–0.84(m,9H);
19F NMR(377MHz,DMSO-d
6)δ-122.27。
步骤11
通过制备级超临界液相色谱法对步骤9所得化合物34-9(700.0毫克)进行手性拆分:手性柱CHIRALPAK IA,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:甲醇/二氯甲烷=2/1(0.1%2摩尔/升氨甲醇);流速:70毫升/分钟;在12分钟内用40%的B相进行洗脱,检测器UV 220/235纳米,得到两个产品。较短保留时间(6.68分钟)的产品为化合物34-9a(两个立体异构体的混合物,黄色油状物,340毫克,回收率48%),化合物34-9a:MS(ESI,m/z):1246.5/1248.5[M+H]
+;较长保留时间(10.32分钟)的产品为化合物34-9b(两个立体异构体的混合物,黄色油状物,280毫克,回收率40%),化合物34-9b:MS(ESI,m/z):1246.5/1248.5[M+H]
+。
步骤12
通过制备级超临界液相色谱法对步骤11所得化合物34-9a(340毫克)进行手性拆分:手性柱CHIRALPAK IA,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:甲醇(0.5%2摩尔/升氨甲醇);流速:70毫升/分钟;在10分钟内用30%的B相进行洗脱,检测器UV 222纳米,得到两个产品。较短保留时间(6.12分钟)的产品为化合物34-9aa(黄色油状物,12毫克,回收率3%),化合物34-9aa:MS(ESI,m/z):1246.5/1248.5[M+H]
+;较长保留时间(6.78分钟)的产品为化合物34-9ab(黄色油状物,290毫克,回收率85%),化合物34-9ab:MS(ESI,m/z):1246.5/1248.5[M+H]
+。
步骤13
通过制备级超临界液相色谱法对步骤11所得化合物34-9b(280毫克)进行手性拆分:手性柱CHIRALPAK IA,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:甲醇(0.5%2摩尔/升氨甲醇);流速:70毫升/分钟;在10分钟内用30%的B相进行洗脱,检测器UV 222纳米,得到两个产品。较短保留时间(4.8分钟)的产品为化合物34-9ba(黄色油状物,160毫克,回收率57%),化合物34-9ba:MS(ESI,m/z):1246.5/1248.5[M+H]
+;较长保留时间(6.03分钟)的产品为化合物34-9bb(黄色油状物,50毫克,回收率17%),化合物34-9bb:MS(ESI,m/z):1246.5/1248.5[M+H]
+。
步骤14
采用步骤10中的方法,以化合物34-9aa为反应起始物料得到化合物34aa(白色固体,6毫克,产率53%)。MS(ESI,m/z):1102.1/1104.1[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.84(s,1H),8.72–8.40(m,1H),7.98–7.92(m,1H),7.74(d,1H),7.47–7.32(m,5H),7.26(d,J=2.4Hz,1H),7.24–7.15(m,2H),7.04–7.01(m,1H),4.68–4.43(m,8H),4.41–4.34(m,1H),4.30–4.23(m,1H),4.12–4.01(m,1H),3.88–3.80(m,1H),3.80–3.61(m,5H),3.60–3.35(m,4H),3.07–2.96(m,1H),2.86–2.73(m,1H),2.48–2.32(m,5H),2.22–2.13(m,2H),2.10–1.99(m,3H),1.99–1.75(m,9H),1.66–1.52(m,2H),0.98(s,9H);
19F NMR(377MHz,CD
3OD)δ-122.92。
步骤15
采用步骤10中的方法,以化合物34-9ab为反应起始物料得到34ab(白色固体,41.2毫克,产率49%)。MS(ESI,m/z):1102.1/1104.1[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ8.96(s,1H),8.32–8.19(m,1H),7.97–7.89(m,1H),7.80(d,J=8.7Hz,2H),7.48–7.34(m,5H),7.29(d,J=2.4Hz,1H),7.26–7.17(m,2H),7.06(d,J=2.3Hz,1H),4.55–4.48(m,1H),4.48–4.29(m,6H),4.26–4.19(m,1H),4.13–3.95(m,2H),3.81–3.69(m,3H),3.68–3.57(m,5H),3.08–2.89(m,2H),2.88–2.79(m,1H),2.49–2.32(m,5H),2.28–2.17(m,2H),2.09–1.95(m,1H),1.95–1.82(m,4H),1.79–1.60(m,9H),1.50–1.42(m,2H),0.89(s,9H);
19F NMR(377MHz,DMSO-d
6)δ-122.21,-122.25。
步骤16
采用步骤10中的方法,以化合物34-9ba为反应起始物料得到34ba(白色固体,37.2毫克,产率50%)。MS(ESI,m/z):1102.1/1104.1[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ8.97(s,1H),8.62–8.56(m,1H),8.24–8.19(m,2H),7.95–7.87(m,2H),7.80(d,J=8.3Hz,1H),7.49–7.35(m,5H),7.28(d,J=2.4Hz,1H),7.24–7.19(m,2H),7.06(d,J=2.4Hz,1H),4.57–4.30(m,8H),4.25–4.18(m,1H),4.11–4.05(m,1H),4.01–3.95(m,1H),3.74–3.70(m,2H),3.59–3.55(m,5H),3.06–3.01(m,1H),2.90–2.78(m,2H),2.43–2.37(m,6H),2.23–2.14(m,1H),2.07–1.98(m,1H),1.93–1.78(m,4H),1.77–1.42(m,11H),0.91(s,9H);
19F NMR(377MHz,DMSO-d
6)δ-122.29。
步骤17
采用步骤10中的方法,以化合物34-9bb为反应起始物料得到34bb(白色固体,18毫克,产率38%)。MS(ESI,m/z):1102.1/1104.1[M+H]
+;
1H NMR(400MHz,CD
3OD)δ8.85(s,1H),8.53(s,1H),7.95(d,J=1.7Hz,1H),7.74(d,J=8.3Hz,1H),7.48–7.33(m,5H),7.26(d,J=2.4Hz,1H),7.22–7.14(m,2H),7.07–6.98(m,1H),4.71–4.59(m,3H),4.57–4.43(m,5H),4.39–4.32(m,1H),4.27–4.18(m,1H),4.16–4.07(m,1H),3.88–3.46(m,9H),3.07–2.87(m,2H),2.49–2.33(m,5H),2.28–2.14(m,2H),2.13–1.94(m,6H),1.94–1.75(m,6H),1.65–1.46(m,2H),1.36–1.22(m,1H),1.07–0.93(m,8H),0.92–0.82(m,1H);
19F NMR(377MHz,CD
3OD)δ-122.84。
实施例35
(2S,4R)-1-((S)-2-(2-((S)-2-(((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐35
步骤1:
在25摄氏度搅拌条件下,向L-脯氨醇(3.00克,29.6毫摩尔,1.00当量)和烯丙基氯(2.27克,29.6毫摩尔,1.00当量)的乙腈(30毫升)溶液中加入碳酸钾(8.20克,59.3毫摩尔,2.00当量)。在氮气保护搅拌条件下,混合物在80摄氏度下反应16小时,反应过程通过液质和薄层层析来监控。反应完成后,降温至25摄氏度,过滤除去不溶物,滤饼用乙腈(10毫升 x 3)洗涤,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物35-1(无色油状物,2.84克,产率64%)。MS(ESI,m/z):142.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ5.96–5.82(m,1H),5.24–5.14(m,1H),5.14–5.06(m,1H),3.67–3.59(m,1H),3.46–3.36(m,2H),3.16–3.07(m,1H),2.98–2.88(m,1H),2.81(s,1H),2.69–2.59(m,1H),2.37–2.26(m,1H),1.97–1.83(m,1H),1.83–1.73(m,1H),1.77–1.66(m,2H)。
步骤2
在零摄氏度,氮气保护搅拌条件下,向化合物16-3a(250毫克,0.387毫摩尔,1.0当量)和化合物35-1(57.5毫克,0.387毫摩尔,1当量)的四氢呋喃(3毫升)溶液中滴加叔丁醇钠的四氢呋喃溶液(2摩尔/升,194毫升,0.387毫摩尔,1当量)。所得混合物在零摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入10毫升水淬灭反应,然后用乙酸乙酯(10毫升 x 3)萃取混合液,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所 得馏分通过减压旋蒸除去溶剂,得到化合物35-2(黄色油,175毫克,产率59%)。MS(ESI,m/z):718.2/720.2[M+H]
+。
步骤3
在25摄氏度氮气保护条件下,依次向50毫升三口瓶中加入化合物35-2(90毫克,0.12毫摩尔,1.0当量),1,3-二甲基巴比妥酸(29毫克,0.178毫摩尔,1.5当量),四(三苯基膦)钯(7.24毫克,0.006毫摩尔,0.05当量)和二氯甲烷(2毫升)。所得混合物在25摄氏度反应5小时,反应过程通过液质和薄色谱层层析监控。反应结束后减压浓缩除去多余的溶剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→50%的乙腈/水流动相(0.1%甲酸)进行洗脱,检测器UV254纳米,得到化合物35-3(橙色固体,74毫克,产率85%)。MS(ESI,m/z):678.3/680.3[M+H]
+。
步骤4
在0摄氏度搅拌条件下,向化合物35-3(74.0毫克,0.10毫摩尔,1.00当量),醋酸(13.1毫克,0.21毫摩尔,2.00当量)和乙醛酸乙酯(16.7毫克,0.15毫摩尔,1.50当量)的甲醇(4.0毫升)溶液中加入氰基硼氢化钠(10.3毫克,0.15毫摩尔,1.50当量)。混合物在25摄氏度搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应完全后,混合物通过减压下浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物35-4(白色固体,40毫克,产率51%)。MS(ESI,m/z):764.3/766.3[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向化合物35-4(40毫克,0.031毫摩尔,1.00当量)的四氢呋喃/甲醇/水的混合溶液(3/1/1,1毫升)中加入二水合氢氧化锂(2.77毫克,0.062毫摩尔,2.00当量)。混合物在25摄氏度搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应完全后,混合物通过减压下浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→95%的乙腈/水流动相(0.1%三氟乙酸)进行洗脱;检测器:UV254/220纳米;得到化合物35-5(白色固体,15毫克,产率61%)。MS(ESI,m/z):736.3/738.3[M+H]
+。
步骤6
在25摄氏度搅拌条件下,向化合物35-5(14毫克,0.015毫摩尔,1.00当量)的N,N-二甲基甲酰胺(1.00毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(9.13毫克,0.022毫摩尔,1.50当量)。混合物在25摄氏度搅拌反应20分钟。然后向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(8.27毫克,0.018毫摩尔,1.20当量)和N,N-二异丙基乙胺(8.28毫克,0.060毫摩尔,4.00当量)。混合物在25摄氏度下继续搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,混合物通过制备级高效液相色谱法纯化:色谱柱XBridge Prep C18 OBD Column,19 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;在8分钟内用54%→80%的B相进行洗脱,检测器UV 225/278纳米。得到化合物35-6(白色固体,14毫克,产率76%)。MS(ESI,m/z):1148.6/1150.6[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物35-6(14毫克,0.012毫摩尔,1.00当量)的甲醇(1毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,1毫升)。反应液在25摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过反相色谱(C18柱)进行纯化,在10分钟内用10%→60%的乙腈/水(0.1%盐酸)流动相进行洗脱;检测器:UV 254/220纳米;得到化合物35(黄色固体,9毫克,产率69%)。MS(ESI,m/z):1004.6/1006.1[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ9.04–8.98(m,1H),8.78–8.71(m,1H),8.03–7.96(m,1H),7.81(d,J=8.5Hz,1H),7.48–7.34(m,5H),7.31(d,J=2.4Hz,1H),7.26–7.14(m,2H), 7.09–7.04(m,1H),4.79–4.62(m,2H),4.61–4.47(m,3H),4.45–4.28(m,4H),4.27–4.11(m,4H),4.09–3.98(m,1H),3.96–3.85(m,2H),3.79–3.64(m,2H),3.30–3.17(m,1H),2.43(s,3H),2.32–2.19(m,1H),2.10–1.82(m,9H),1.30–1.13(m,2H),0.98–0.72(m,10H);
19F NMR(282MHz,DMSO-d
6)δ-121.85。
实施例36
((3S或3R,7aS或7a R)-7a-(((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)六氢-1H-吡咯啉-3-基)甲基(2-(3-((S)-1-((2S,4R)-4-羟基-2-((4-(4-甲基噻唑-5-基)苄基)氨甲酰基)吡咯烷-1-基)-3,3-二甲基-1-氧代丁烷-2-基)氨基)-3-氧代丙氧基)乙基)(甲基)氨基甲酸酯二甲酸盐36
步骤1
在25摄氏度搅拌条件下,向500毫升圆底烧瓶中依次加入N-甲基-2-羟基乙胺(10.0克,126.48毫摩尔,1.0当量),二氯甲烷(60.0毫升),三乙胺(55.52毫升,379.44毫摩尔,3.0当量)和二碳酸二叔丁酯(30.51克,132.80毫摩尔,1.05当量)。混合物在25摄氏度下搅拌3小时。反应过程通过液质和薄层层析来监控。反应完成后,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-1(黄色油,17.0克,产率73%)。MS(ESI,m/z):176.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ3.77–3.73(m,2H),3.42–3.38(m,2H),2.93(s,3H),1.47(s,9H)。
步骤2
在0摄氏度,氮气保护搅拌条件下,向化合物36-1(17.0克,92.2毫摩尔,1.0当量),4-二甲氨基吡啶(1.2克,9.2毫摩尔,0.1当量)和三乙胺(19.6克,184.3毫摩尔,2.0当量)的二氯甲烷(200.0毫升)溶液中加入对甲苯磺酰氯(20.3克,101.4毫摩尔,1.1当量)。所得混合物在25摄氏度下搅拌 反应16小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→60%甲基叔丁基醚/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-2(黄色固体,9.9克,产率26%)。MS(ESI,m/z):330.0[M+H]
+。
步骤3
在0摄氏度,氮气保护搅拌条件下,向3-丁烯-1-醇(1.8克,23.8毫摩尔,1.1当量)的N-N二甲基甲酰胺(10.0毫升)溶液中加入氢化钠(60%)(1.1克,28.56毫摩尔,1.2当量)。所得混合物在0摄氏度,氮气保护条件下搅拌30分钟。然后向反应液中加入36-2(8.9克,21.6毫摩尔,1.0当量)。混合物在25摄氏度下反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,加入300毫升水淬灭反应,水层用乙酸乙酯(300毫升 x 3)萃取。合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂;滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→40%甲基叔丁基醚/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-3(无色油,570毫克,产率10.9%)。MS(ESI,m/z):230.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ5.86–5.76(m,1H),5.13–5.01(m,2H),3.55–3.47(m,4H),3.39–3.36(m,2H),2.91(s,3H),2.36–2.29(m,2H),1.46(s,9H)。
步骤4
在0摄氏度搅拌条件下,向化合物36-3(500.0毫克,2.1毫摩尔,1.0当量)的甲醇(3.0毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,3.0毫升)。所得混合物在25摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩除去多余的溶剂,得到化合物36-4(黄色液体,330.0毫克,产率96%)。MS(ESI,m/z):130.2[M+H]
+。
步骤5
在0摄氏度氮气保护搅拌条件下,向N-苄氧羰基-L-脯氨酸(20克,76.22毫摩尔,1.0当量)的甲醇(200.0毫升)溶液中逐滴加入氯化亚砜(27.48克,228.67毫摩尔,3.0当量)。混合物在0摄氏度下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→30%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-5(无色油,20克,产率94%)。MS(ESI,m/z):264.3[M+H]
+;
1H NMR (300MHz,CDCl
3)δ7.40–7.28(m,5H),5.22–5.00(m,2H),4.41–4.33(m,1H),3.80–3.41(m,5H),2.31–2.13(m,1H),2.09–1.73(m,3H)。
步骤6
在-78摄氏度氮气保护搅拌条件下,向化合物36-5(20克,72.17毫摩尔,1.0当量)的四氢呋喃(200.0毫升)溶液中依次缓慢加入和双三甲基硅基胺基锂(1摩尔/升的四氢呋喃溶液,144毫升,144毫摩尔,2.0当量)和4-溴-1-丁烯(12.3克,80.604毫摩尔,1.2当量)。混合物在25摄氏度下搅拌反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液中加入水(300毫升)淬灭反应,然后用乙酸乙酯(300毫升 x 3)萃取,合并有机相,有机相用饱和食盐水(80毫升 x 3)洗涤,用无水硫酸钠干燥,过滤,滤液浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→30%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-6(无色油,17克,产率69%)。MS(ESI,m/z):318.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.42–7.29(m,5H),5.90–5.67(m,1H),5.21–4.90(m,4H),3.88–3.67(m,3H),3.57–3.44(m,2H),2.53–2.19(m,1H),2.18–1.79(m,7H)。
步骤7
在25摄氏度氮气保护搅拌条件下,向化合物36-6(17克,50.884毫摩尔,1.0当量)的二氯甲烷(200.0毫升)溶液中缓慢加入间氯过氧苯甲酸(23.12克,127.21毫摩尔,2.5当量)。混合物在25摄氏度下搅拌反应5小时,反应过程通过薄层层析来监控(乙酸乙酯/石油醚=1/4,R
f=0.2)。反应完全后,向反应液中加入饱和硫代硫酸钠水溶液(300.0毫升)淬灭反应,然后用二氯甲烷(300毫升x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤,滤液浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→50%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-7(棕黄色油,12克,产率66%)。MS(ESI,m/z):334.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.40–7.31(m,5H),5.20–5.06(m,2H),3.86–3.71(m,3H),3.55–3.44(m,2H),2.98–2.68(m,2H),2.56–2.23(m,2H),2.19–1.81(m,5H),1.72–1.35(m,2H)。
步骤8
在25摄氏度氮气保护搅拌条件下,向化合物36-7(12克,34.195毫摩尔,1.0当量)的甲醇(120.0毫升)溶液中加入钯/碳(10%钯含量,1.2克)。通过置换气操作将氮气置换成氢气(1.5大气压)。混合物在25摄氏度氢气氛围下搅拌反应5小时,反应过程通过液质和薄层层析来监控。反应结束后,过滤除去不溶物,滤饼用甲醇(100毫升)清洗,滤液减压浓缩得到化合物36-8的粗产品。(无色油,7克,产率97%)。MS(ESI,m/z):200.1[M+H]
+。
步骤9
在0摄氏度氮气保护搅拌条件下,向化合物36-8(7克,33.375毫摩尔,1.0当量)和咪唑(9.09克,126.845毫摩尔,3.8当量)的N,N-二甲基甲酰胺(70.0毫升)溶液中缓慢加入叔丁基二苯基氯硅烷(19.31克,66.75毫摩尔,2.0当量)。混合物在25摄氏度下搅拌反应16小时,反应过程通过薄层层析(乙酸乙酯/石油醚=1/3,R
f=0.5,0.2)来监控。反应完全后,向反应液中加入水(500毫升)淬灭反应,然后用乙酸乙酯(500毫升 x 3)萃取,合并有机相,有机相用饱和食盐水(80毫升 x 3)洗涤,用无水硫酸钠干燥,过滤,滤液浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→40%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,分别得到两个化合物。极性相对较小的化合物为36-8a(无色油,6克,产率38%),极性相对较大的化合物为36-8b(无色油,5.7克,产率37%)。
化合物36-8b:MS(ESI,m/z):438.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.74–7.63(m,4H),7.51–7.35(m,6H),4.03–3.96(m,1H),3.88–3.81(m,1H),3.74(s,3H),3.46–3.35(m,1H),3.01–2.79(m,2H),2.55–2.45(m,1H),2.30–2.15(m,1H),2.04–1.75(m,5H),1.70–1.53(m,1H),1.07(s,9H)。
步骤10
在-20摄氏度氮气保护搅拌条件下,向氢化铝锂(1.46克,36.468毫摩尔,3.0当量)的四氢呋喃 (56毫升)溶液中缓慢加入化合物36-8b(5.6克,12.156毫摩尔,1.0当量)。混合物在-20摄氏度下搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,在零摄氏度搅拌条件下,依次向反应液中加入水(1.46毫升),15%的氢氧化钠水溶液(1.46毫升)和水(5.38毫升)。过滤除去不溶物,滤饼用四氢呋喃(20毫升 x 3)清洗,滤液减压浓缩得到化合物36-9b(类白色固体,5.0克,产率95%)。MS(ESI,m/z):410.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.74–7.64(m,4H),7.49–7.33(m,6H),3.97–3.89(m,1H),3.79–3.72(m,1H),3.32–3.22(m,2H),3.22–3.13(m,1H),2.86–2.79(m,1H),2.74–2.65(m,1H),1.99–1.91(m,1H),1.82–1.57(m,6H),1.56–1.47(m,1H),1.06(s,9H)。
步骤11
通过超临界液相色谱对步骤10所得化合物36-9b(5克)进行手性拆分:手性柱(R,R)-WHELK-O1-Kromasil,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:异丙醇/二氯甲烷=2/1(2摩尔/升氨甲醇);流速:70毫升/分钟;在6.5分钟内用55%的B相进行洗脱,检测器UV 204纳米。得到两个产品。较短保留时间(3.27分钟)的产品为化合物36-9ba,((3R或3S,7aR或7aS)-3-((叔丁基二苯基硅氧基)甲基)四氢-1H-吡咯啉-7a(5H)-基)甲醇(无色油,2.2克,回收率44%),MS(ESI,m/z):410.3[M+H]
+;较长保留时间(5.87分钟)的产品为化合物36-9bb,((3S或3R,7aS或7aR)-3-((叔丁基二苯基硅氧基)甲基)四氢-1H-吡咯啉-7a(5H)-基)甲醇(无色油,1.9克,回收率38%),MS(ESI,m/z):410.3[M+H]
+。
步骤12
在0摄氏度氮气保护搅拌条件下,向化合物16-3a(1.36克,2.106毫摩尔,1.00当量)的无水四氢呋喃(10毫升)溶液中依次滴加化合物36-9ba(1.00克,2.317毫摩尔,1.10当量)和叔丁醇钾的四氢呋喃溶液(1.0摩尔/升,2.5毫升,2.527毫摩尔,1.20当量)。反应液在0摄氏度氮气保护下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过硅胶色谱柱进行纯化,用0%→10%的甲醇/二氯甲烷流动相进行洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-10ba(黄色固体,1.09克,产率50%)。MS(ESI,m/z):986.3/988.3[M+H]
+。
步骤13
在25摄氏度搅拌条件下,向化合物36-10ba(1.00克,0.963毫摩尔,1.00当量)的四氢呋喃(8毫升)溶液中滴加四丁基氟化铵(1.59克,5.778毫摩尔,6.00当量)。反应液在25摄氏度下反应3小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过硅胶色谱柱进行纯化,用0%→10%的甲醇/二氯甲烷流动相进行洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-11ba(白色固体,750毫克,产率98%)。MS(ESI,m/z):748.2/750.2[M+H]
+。
步骤14
在25摄氏度氮气保护搅拌条件下,向史莱克管中依次加入化合物36-11ba(200.0毫克,0.25毫摩尔,1.0当量),四氢呋喃(3.0毫升),对硝基苯基氯甲酸酯(107.7克,0.5毫摩尔,2.0当量),化合物36-4(103.6毫克,0.7毫克,3.0当量)以及N-N-二异丙基乙胺(138.1毫克,1.0毫摩尔,4.0当量)。所得混合物在0摄氏度下搅拌2小时,反应过程通过液质和薄层层析来监控。反应完成后,减压浓缩除去多余的试剂得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物36-12ba(白色固体,120.0毫克,产率49%)。MS(ESI,m/z):903.1/905.1[M+H]
+。
步骤15
在0摄氏度,氮气保护搅拌条件下,向化合物36-12ba(110.0毫克,0.1毫摩尔,1.0当量)和高碘酸钠(100.1毫克,0.5毫摩尔,4.0当量)的四氯化碳/乙腈/水的混合溶液(2/2/3,4.5毫升)中加入三氯化钌(1.3毫克,0.005毫摩尔,0.05当量)。混合物在0摄氏度下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得粗产品。所得混合物通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→90%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器,UV254纳米;得到化合物36-13ba(白色固体,55毫克,产率49%)MS(ESI,m/z):921.5/923.5[M+H]
+。
步骤16
在25摄氏度搅拌条件下,向化合物36-13ba(50毫克,0.05毫摩尔,1.0当量)的N,N-二甲基甲酰胺(2.00毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(23.7毫克,0.06毫摩尔,1.2当量)。混合物在25摄氏度搅拌反应20分钟。然后向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(26.9毫克,0.06毫摩尔,1.2当量)和N,N-二异丙基乙胺(26.9毫克,0.2毫摩尔,4.0当量)。混合物在25摄氏度下继续搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,混合物通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→90%的乙腈/水流动相(0.1%三氟乙酸)进行洗脱;检测器UV254纳米;得到化合物36-14ba(白色固体,46毫克,产率63%)。MS(ESI,m/z):1333.0/1335.0[M+H]
+。
步骤17
在25摄氏度搅拌条件下,向化合物36-14ba(45.0毫克,0.03毫摩尔,1.0当量)的甲醇(1毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,1毫升)。混合物在25摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过高效液相色谱进行纯化,色谱柱Sunfire prep C18 column,30 x 150毫米,5微米;流动相A:水(0.05%甲酸),流动相B:乙腈;流速:60毫升/分钟;在7分钟内用10%→35%的B相进行洗脱,检测器UV 220/235纳米,得到化合物36(白色固体,35毫克,产率79%)。MS(ESI,m/z):1189.1/1191.1[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ8.98–8.91(m,1H),8.48–8.34(m,2H),8.03–7.97(m,1H),7.95–7.81(m,2H),7.51–7.34(m,6H),7.32–7.18(m,2H),7.09(d,J=2.4Hz,1H),4.56–4.40(m,7H),4.33–4.21(m,3H),4.02–3.90(m,2H),3.86–3.58(m,7H),3.55–3.46(m,2H),3.43–3.29(m,2H),3.28–3.18(m,1H),3.15–3.05(m,2H),2.91–2.81(m,3H),2.48–2.35(m,5H),2.24–1.78(m,14H),1.26–1.16(m,2H),1.00–0.83(m,9H);
19F NMR(282MHz,DMSO-d
6)δ-122.36。
实施例37
(2S,4R)-1-((S)-2-(3-(3-((3R,5S)-5-(((S或R)-4-((1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)-1-甲基吡咯烷-3-基)氧基)丙酰 胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺二盐酸盐37
步骤1
在0摄氏度氮气保护搅拌条件下,向(2S,4R)-1-(叔丁氧羰基)-4-羟基-2-(羟甲基)吡咯烷(10.0克,43.7毫摩尔,1.0当量)和咪唑(3.3克,45.9毫摩尔,1.1当量)的N,N-二甲基甲酰胺(100毫升)溶液中缓慢加入叔丁基二苯基氯硅烷(13.3克,45.9毫摩尔,1.1当量)。混合物在25摄氏度搅拌条件下反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,在0摄氏度条件加入600毫升水稀释反应,然后用乙酸乙酯(600毫升 x 3)萃取,合并有机相,有机相用饱和食盐水清洗(200毫升 x 3),再用无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→40%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物37-1(白色固体,10.5克,产率52%)。MS(ESI,m/z):456.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.69–7.62(m,4H),7.48–7.34(m,6H),4.59–4.51(m,1H),4.12–4.01(m,1H),4.01–3.77(m,1H),3.73–3.64(m,1H),3.63–3.47(m,2H),2.40–2.29(m,1H),2.13–2.00(m,1H),1.87–1.69(m,1H),1.41(s,9H),1.06(s,9H)。
步骤2
在0摄氏度氮气保护搅拌条件下,向化合物37-1(5.65克,11.8毫摩尔,1.5当量)的N,N-二甲基甲酰胺(30毫升)溶液中加入氢化钠(60%,0.5克,11.8毫摩尔,1.5当量)。混合物在0摄氏度下搅拌0.5小时后,加入化合物25-3(2.4克,7.9毫摩尔,1.0当量)。反应液在25摄氏度条件下反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,在0摄氏度下,向反应液中加入200毫升水淬灭反应。水相用乙酸乙酯(200毫升 x 3)萃取,合并后有机相用饱和食盐水(80毫升 x 3) 清洗,无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水流动相(0.1%三氟乙酸)进行洗脱;检测器UV254纳米;得到化合物37-2(无色油状物,1.7克,产率36%)。MS(ESI,m/z):568.3[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.67–7.60(m,4H),7.45–7.31(m,6H),5.92–5.76(m,1H),5.15–4.92(m,2H),4.15–3.90(m,2H),3.73–3.34(m,10H),2.39–2.20(m,3H),2.15–1.95(m,1H),1.88–1.76(m,2H),1.48–1.30(m,9H),1.05(s,9H)。
步骤3
在25摄氏度搅拌条件下,将化合物37-2(1.7克,2.8毫摩尔,1.0当量)溶于四氢呋喃(17毫升)中,然后加入四丁基氟化铵(1.5克,5.5毫摩尔,2.0当量)。反应液在25摄氏度搅拌条件下反应1.5小时,反应过程通过液质和薄层层析来监控。反应完全后,加入50毫升水稀释反应,水相用乙酸乙酯(50毫升 x 3)萃取,合并有机相,再用无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物37-3(无色油状物,900毫克,产率94%)。MS(ESI,m/z):330.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ5.88–5.76(m,1H),5.13–4.97(m,2H),4.10–4.01(m,1H),3.96–3.88(m,1H),3.73–3.67(m,1H),3.61–3.54(m,2H),3.53–3.44(m,6H),3.41–3.35(m,1H),2.37–2.28(m,2H),2.14–2.06(m,1H),1.85–1.77(m,2H),1.74–1.60(m,1H),1.47(s,9H)。
步骤4
在0摄氏度氮气保护搅拌条件下,向化合物37-3(850毫克,2.5毫摩尔,1.0当量)的无水四氢呋喃(9毫升)溶液中缓慢加入四氢铝锂(195.9毫克,4.9毫摩尔,2.0当量)。反应液在55摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,在0摄氏度下,向反应液中依次加入水(195.9毫克),15%的氢氧化钠水溶液(195.9毫克),水(588毫克)。过滤除去不溶物,滤饼用四氢呋喃(15毫升 x 3)清洗,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物37-4(无色油状物,430毫克,产率72%)。MS(ESI,m/z):244.2[M+H]
+。
步骤5
在-20摄氏度氮气保护条件下,向化合物16-3a(250毫克,0.39毫摩尔,1.0当量)和化合物37-4(119.0毫克,0.46毫摩尔,1.2当量)的无水四氢呋喃(3毫升)溶液中加入叔丁醇钾的四氢呋喃溶液(1摩尔/升,0.46毫升,0.46毫摩尔,1.2当量)。混合物在-20摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,向反应液加入10毫升水淬灭反应,然后用乙酸乙酯(15毫升 x 3)萃取,合并有机相并用无水硫酸钠干燥,过滤除去不溶物,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物37-5(白色固体,150毫克,产率45%)。MS(ESI,m/z):820.3/822.3[M+H]
+。
步骤6
在25摄氏度搅拌条件下,将化合物37-5(160毫克,0.19毫摩尔,1.0当量)溶于1.7毫升的四氯化碳和1.7毫升的乙腈中,然后加入高碘酸钠(250.3毫克,1.1毫摩尔,6.0当量)。将体系降温至0摄氏度,然后缓慢滴加三氯化钌(2.2毫克,0.01毫摩尔,0.05当量)的水溶液(2.5毫升)。反应液在0摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液倒入15毫升水中,水相用三氯甲烷/异丙醇的混合溶液(3:1,20毫升 x 3)萃取,合并有机相并用无水硫酸钠干燥。过滤除去不溶物,有机相减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器UV254纳米;得到化合物37-6(白色固体,60毫克,产率38%)。MS(ESI,m/z):838.4/840.4[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物37-6(60毫克,0.07毫摩尔,1.0当量)的N,N-二甲基甲酰胺(3毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(32.7毫克,0.08毫 摩尔,1.2当量)。混合物在25摄氏度下反应15分钟。然后向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(44.4毫克,0.08毫摩尔,1.2当量)和N,N-二异丙基乙胺(46.3毫克,0.34毫摩尔,5.0当量)。反应液在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,混合物通过高效液相色谱进行纯化,色谱柱XSelect CSH Prep C18 OBD,19 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;在8分钟内用56%→80%的B相进行洗脱,检测器UV 220/235纳米。得到化合物37-7(无色油状物,53毫克,产率59%)。MS(ESI,m/z):1264.5/1266.5[M+H]
+。
步骤8
在0摄氏度搅拌条件下,向化合物37-7(53毫克,0.04毫摩尔,1.0当量)的甲醇(2毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,2毫升)。混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的甲醇/水流动相(0.1%盐酸)进行洗脱;检测器,UV220纳米;得到化合物37(黄色固体,32.3毫克,收率64%)。MS(ESI,m/z):1120.3/1122.3[M+H]
+。
1H NMR(400MHz,DMSO-d
6)δ11.36–11.14(m,1H),10.17–9.97(m,1H),9.92–9.73(m,1H),9.14–9.06(m,1H),8.45(d,J=7.8Hz,1H),8.04–7.98(m,1H),7.91–7.78(m,2H),7.48–7.41(m,3H),7.41–7.35(m,2H),7.33–7.28(m,1H),7.26–7.16(m,2H),7.14–7.09(m,1H),4.97–4.84(m,1H),4.84–4.74(m,2H),4.62–4.39(m,4H),4.30–4.25(m,1H),4.24–4.11(m,3H),4.04–3.91(m,3H),3.87–3.77(m,1H),3.66–3.51(m,4H),3.49–3.34(m,5H),3.22–3.13(m,1H),3.03–2.92(m,3H),2.46(s,3H),2.41–2.28(m,2H),2.09–1.85(m,6H),1.82–1.63(m,3H),1.51–1.31(m,3H),1.01–0.80(m,9H);
19F NMR(377MHz,DMSO-d
6)δ-121.95。
实施例38
(2S,4R)-1-((S)-2-(3-(3-((S)-2-((4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-8-氟-7-(3-羟基萘-1-基)吡啶[4,3-d]嘧啶-2-基)氧基)甲基)吡咯烷-1-基)丙氧基)丙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-甲基噻唑-5-苄基)吡咯烷-2-甲酰胺二甲酸盐38
步骤1:
在25摄氏度搅拌条件下,向2-氯-3-氟-4-氨基吡啶(1克,6.82毫摩尔,1.0当量)的乙腈(10毫升)溶液中加入N-碘代丁二酰亚胺(1.84克,7.77毫摩尔,1.2当量)和对甲苯磺酸(130毫克,0.65毫摩尔,0.1当量)。混合物在70摄氏度条件下反应16小时。反应过程通过液质和薄层层析来监控。反应结束后,混合物冷却至25摄氏度,加入30毫升水稀释,用乙酸乙酯(50毫升 x 3)萃取,合并有机相。有机相依次用50毫升饱和碳酸钠溶液,50毫升饱和亚硫酸钠溶液以及50毫升饱和食盐水洗涤,有机相再用无水硫酸钠干燥,过滤,滤液浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→45%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物38-1(黄色固体,1.72克,产率95%)。MS(ESI,m/z):272.9/274.9[M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.17(s,1H),4.84(s,2H)。
步骤2:
在25摄氏度氮气保护搅拌条件下,向化合物38-1(1克,3.67毫摩尔,1.0当量)的乙醇(10毫升)溶液中加入三乙胺(1.38克,12.96毫摩尔,3.6当量)和二氯二(三苯基膦)钯(266毫克,0.36毫摩尔,0.1当量)。混合物在一氧化碳氛围中80摄氏度条件下反应15小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至25摄氏度,用硅藻土过滤,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→31%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物38-2(黄色固体,660毫克,产率74%)。MS(ESI, m/z):219.3/221.3[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.56(s,1H),4.41(q,J=7.1Hz,2H),1.43(t,J=7.1,3H)。
步骤3:
在25摄氏度氮气保护搅拌条件下,向化合物38-2(660毫克,2.87毫摩尔,1.0当量)的无水四氢呋喃(6毫升)溶液中滴加三氯乙酰异氰酸酯(853毫克,4.30毫摩尔,1.5当量)。混合物在25摄氏度下反应20分钟,反应过程通过液质和薄层层析来监控。反应结束后,将反应液浓缩得到粗产品。所得粗产品用甲基叔丁基醚(10毫升)打浆,过滤,滤饼用甲基叔丁基醚(2毫升 x 3)洗涤,滤饼干燥得到化合物38-3(白色固体,1.0克,产率77%)。MS(ESI,m/z):406.0/408.0/410.0[M+H]
+;
1H NMR(400MHz,CDCl
3)δ11.19(s,1H),8.91(s,1H),8.78(d,J=0.8Hz,1H),4.48(q,J=7.1Hz,2H),1.43(t,J=7.1,3H)。
步骤4:
在25摄氏度搅拌下,向化合物38-3(1克,2.334毫摩尔,1.00当量)的甲醇(10毫升)溶液中滴加氨甲醇溶液(7摩尔每升,1毫升)。混合液在25摄氏度条件下反应1小时。反应过程通过液质和薄层层析来监控。反应结束后,将反应液浓缩得到粗产品。所得粗产品通过甲基叔丁基醚(10毫升)打浆,过滤,滤饼用甲基叔丁基醚(2毫升 x 3)洗涤,滤饼干燥得到化合物38-4(白色固体,594毫克,产率94%)。MS(ESI,m/z):216.1/218.1[M+H]
+。
1H NMR(400MHz,DMSO-d
6)δ8.34(d,J=1.2Hz,1H)。
步骤5:
在0摄氏度氮气保护的条件下,向干燥的100毫升单口瓶中依次加入化合物38-4(500毫克,2.20毫摩尔,1.0当量),三氯氧磷(9毫升)和N,N-二异丙基乙胺(0.9毫升)。混合物在0摄氏度条件下搅拌10分钟,然后在90摄氏度下反应12小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→30%乙酸乙酯/石油醚梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物38-5产品(黄色固体,425毫克,产率72%)。
步骤6:
在25摄氏度氮气保护搅拌条件下,将化合物38-5(425毫克,1.68毫摩尔,1.0当量)溶解在5毫升二氯甲烷中。向该溶液中依次加入N,N-二异丙基乙胺(652毫克,4.80毫摩尔,3.0当量)和(1R,5S)-3,8-二氮杂二环[3.2.1]辛烷-8-甲酸叔丁酯(357毫克,1.60毫摩尔,1.0当量),然后在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→30%乙酸乙酯/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物38-6(黄色固体,700毫克,产率97%)。MS(ESI,m/z):428.2/430.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ8.86(d,J=0.6Hz,1H),4.59–4.41(m,4H),3.78–3.71(m,2H),2.04–1.96(m,2H),1.75–1.65(m,2H),1.54(s,9H)。
步骤7
在0摄氏度搅拌条件下,向化合物38-6(700毫克,1.64摩尔,1.0当量)和化合物25-4(419毫克,1.96毫摩尔,1.2当量)的无水四氢呋喃(10.0毫升)溶液中加入叔丁醇钾的四氢呋喃溶液(1摩尔/升,1.96毫升,1.96毫摩尔,1.2当量)。所得混合物在0摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完成后,加入50毫升水淬灭反应,水相用乙酸乙酯(50毫升 x 3)萃取,合并有机相,有机相用50毫升饱和食盐水洗涤,洗涤后有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物38-7(无色油,570毫克,产率57%)。MS (ESI,m/z):605.4/607.4[M+H]
+。
步骤8
在25摄氏度,氮气保护搅拌条件下,向反应瓶中依次加入化合物38-7(550毫克,0.86毫摩尔,1.0当量),4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)萘-2-醇(368.28毫克,1.29毫摩尔,1.5当量),碳酸铯(592.25毫克,1.73毫摩尔,2.0当量),四三苯基磷钯(105.02毫克,0.086毫摩尔,0.1当量),1,4-二氧六环(5.0毫升)和水(1.0毫升)。混合物在100摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应完成后,向反应液中加入50毫升稀释,水相用乙酸乙酯(50毫升 x 3)萃取。合并有机相,有机相用50毫升饱和食盐水洗涤,无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→60%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物38-8(白色固体,550毫克,产率85%)。MS(ESI,m/z):713.1[M+H]
+。
步骤9
在0摄氏度,氮气保护搅拌条件下,向化合物38-8(480毫克,0.64毫摩尔,1.0当量)和咪唑(137.52毫克,1.92毫摩尔,3.0当量)的二氯甲烷(5毫升)溶液中加入叔丁基二甲基氯硅烷(152.23毫克,0.96毫摩尔,1.5当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应完成后,向反应液中加入50毫升水,二氯甲烷(50毫升 x 3)萃取。合并后有机相用50毫升饱和食盐水洗涤,无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→80%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物38-9(白色固体,420毫克,产率75%)。MS(ESI,m/z):827.4[M+H]
+。
步骤10
在0摄氏度搅拌条件下,向化合物38-9(180毫克,0.21毫摩尔,1.0当量)和高碘酸钠(279.28毫克,1.24毫摩尔,6.0当量)的乙腈(1.0毫升),水(1.5毫升)和四氯化碳(1.0毫升)的混合溶液中加入氯化钌的一水合物(2.45毫克,0.01毫摩尔,0.05当量)。所得混合物在0摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩除去多余的溶剂得粗产物。所得混合物通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→95%的乙腈/水流动相(0.1%氨水,氨水中氨含量25%~28%)进行洗脱;检测器,UV254纳米;得到化合物38-10(黄色油,86毫克,产率47%)。MS(ESI,m/z):845.6[M+H]
+。
步骤11
在25摄氏度搅拌条件下,向38-10(81毫克,0.091毫摩尔,1.0当量)和2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(43.73毫克,0.11毫摩尔,1.2当量)的N,N-二甲基甲酰胺(1毫升)溶液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(53.72毫克,0.11毫摩尔,1.2当量)和N,N-二异丙基乙胺(49.55毫克,0.364毫摩尔,4.0当量)。所得混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,所得混合物通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的[乙腈/甲醇(3/1)]/水流动相(0.1%氨水,氨水中氨含量25%~28%)进行洗脱;检测器UV254纳米;得到化合物38-11(黄色固体,82毫克,产率68%)。MS(ESI,m/z):1257.8[M+H]
+。
步骤12
在25摄氏度,氮气保护搅拌条件下,向化合物38-11(82毫克,0.062毫摩尔,1.0当量)的四氢呋喃(3毫升)溶液中加入四丁基氟化铵(51.14毫克,0.186毫摩尔,3.0当量)。混合物在25摄氏度下搅拌1小时。反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产物。粗产物通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的[乙腈/甲醇(1/1)]/水流动相(0.1%氨水,氨水中氨含量25%~28%)进行洗脱;检测器UV254纳米;得到化合物38-12(黄色固体,42毫克,产率59%)。MS(ESI,m/z):1143.7[M+H]
+。
步骤13
在25摄氏度搅拌条件下,向化合物38-12(40毫克,0.33毫摩尔,1.0当量)的二氯甲烷(2毫升)溶液中加入三氟乙酸(1毫升)。混合物在25摄氏度下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产物。粗产物通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的[乙腈/甲醇(1/1)]/水流动相(0.1%甲酸)进行洗脱;检测器UV254纳米;得到化合物38(白色固体,25毫克,产率63%)。MS(ESI,m/z):1043.5[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ9.16(s,1H),8.97(s,1H),8.59–8.53(m,1H),8.20(s,2H),7.86(d,J=9.4Hz,1H),7.79(d,J=8.3Hz,1H),7.55(d,J=8.4Hz,1H),7.46–7.34(m,5H),7.30–7.21(m,3H),4.55–4.32(m,7H),4.25–4.18(m,1H),4.15–4.09(m,1H),3.71–3.47(m,9H),3.42–3.34(m,2H),3.07–3.01(m,1H),2.96–2.78(m,2H),2.59–2.52(m,1H),2.43(s,3H),2.40–2.25(m,2H),2.22–2.14(m,1H),2.07–1.99(m,1H),1.93–1.85(m,2H),1.74–1.61(m,9H),0.89(s,9H);
19F NMR(377MHz,DMSO-d
6)δ-139.66。
实施例39
(2S,4R)-1-((S)-2-(2-(4-(2-((S)-2-(((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基乙基)-1H-1,2,3-三唑-1-基)乙酰胺基-3-二甲基丁酰基)-4-羟基-4-苄基-4-噻唑-2-甲酰胺-2-苄基-甲酰胺二盐酸盐39
步骤1
在25摄氏度,搅拌条件下,向2-叠氮基乙酸乙酯(5克,36.788毫摩尔,1当量)的水(25.0毫升)/叔丁醇(25.0毫升)混合溶液中加入L-抗坏血酸钠(1.70克,8.093毫摩尔,0.22当量),硫酸铜(0.68克,4.047毫摩尔,0.11当量)和3-丁炔-1-醇(4.07克,55.182毫摩尔,1.5当量)。混合物在25摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,在25摄氏度下加入100毫升水淬灭反应,水相用氯仿/异丙醇(3/1,100毫升 x 3)萃取。合并后有机层用400毫升饱和氯化钠溶液洗涤,洗涤后有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得粗产品。粗产品通过硅胶柱层析法纯化,用0%→7%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物39-1(白色油,5.04克,65%)。MS(ESI,m/z):200.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.56(s,1H),5.14(s,2H),4.26(q,J=7.1Hz,2H),3.92(t,J=6.0Hz,2H),3.07(s,1H),2.97(t,J=6.0Hz,2H),1.30(t,J=7.1Hz,3H)。
步骤2
在0摄氏度,氮气保护搅拌条件下,向化合物39-1(2克,9.538毫摩尔,1.00当量),三乙胺(3.05克,28.614毫摩尔,3当量)和4-二甲氨基吡啶(0.12克,0.954毫摩尔,0.1当量)的二氯甲烷(30.0毫升)溶液中加入对甲苯磺酰氯(2.23克,11.446毫摩尔,1.2当量)。反应液在25摄氏度条件下搅拌反应16小时,反应过程通过液质和薄层层析来监控。反应完全后,在0摄氏度下加入50毫升水淬灭反应,水相用二氯甲烷(50毫升 x 3)萃取,合并后有机相用无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得粗产品。粗产品通过硅胶柱层析法纯化,用0%→30%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物39-2(黄色油,2.98克,产率83%)。MS(ESI,m/z):354.0[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.79–7.71(m,2H),7.57(s,1H),7.33(d,J=8.1Hz,2H),5.13(s,2H),4.33–4.23(m,4H),3.13(t,J=6.5Hz,2H),2.45(s,3H),1.31(t,J=7.1Hz,3H)。
步骤3
在25摄氏度搅拌条件下,向化合物39-2(1克,2.68毫摩尔,1.0当量)和L-脯氨醇(314.85毫克,2.957毫摩尔,1.10当量)的乙腈(5.0毫升)溶液中加入碳酸钾(782.17毫克,5.376毫摩尔, 2.0当量)。混合物在60摄氏度条件下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应结束后过滤除去不溶物,滤饼用乙腈(50毫升 x 3)清洗,滤液减压浓缩得粗产品。粗产品通过硅胶柱层析法纯化,用0%→7%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物39-3(白色固体,320毫克,产率40%)。MS(ESI,m/z):283.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.51(s,1H),5.13(s,2H),4.31–4.21(m,2H),3.70–3.58(m,1H),3.46–3.36(m,1H),3.35–3.24(m,1H),3.23–3.12(m,1H),3.09–2.94(m,2H),2.80–2.67(m,2H),2.46–2.32(m,1H),1.97–1.85(m,1H),1.83–1.73(m,3H),1.30(t,J=7.1Hz,3H)。
步骤4
在-15摄氏度,氮气保护搅拌条件下,向化合物16-3a(300毫克,0.464毫摩尔,1.0当量)和化合物39-3(151.9毫克,0.511毫摩尔,1.1当量)的二氯甲烷(5.0毫升)溶液中加入叔丁醇钾的四氢呋喃溶液(1摩尔每升,1.16毫升,1.16毫摩尔,2.5当量)。反应液在-15摄氏度条件下搅拌反应4小时,反应过程通过液质和薄层层析来监控。反应完全后,向体系中加入2毫升水淬灭反应,减压浓缩得到粗产品,所得粗产品直接通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用30%→80%的(乙腈/甲醇=1/1)/水溶液流动相(0.1%碳酸氢铵)进行洗脱;检测器UV220/254纳米;得到化合物39-4(白色固体,112.0毫克,产率27%)。MS(ESI,m/z):831.4/833.4[M+H]
+。
步骤5
在室温搅拌条件下,向化合物39-4(100.0毫克,0.11毫摩尔,1.0当量)的N,N-二甲基甲酰胺(1.5毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(54.88毫克,0.137毫摩尔,1.2当量)。混合物在25摄氏度下继续搅拌反应10分钟,再向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(61.80毫克,0.125毫摩尔,1.1当量)和N,N-二异丙基乙胺(62.19毫克,0.456毫摩尔,4当量)。混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,所得混合物通过制备级高效液相色谱进行纯化,色谱柱XBridge Prep C18 OBD Column 19 x 150毫米,5微米;流动相A:水(0.05%碳酸氢铵),流动相B:甲醇;流速:25毫升/分钟;在6分钟内用83%→84%的B相进行洗脱,检测器UV 220纳米。得到化合物39-5(白色固体,85毫克,产率55%)。MS(ESI,m/z):1243.1/1245.1[M+H]
+。
步骤6
在0摄氏度搅拌条件下,向化合物39-5(75.0毫克,0.06毫摩尔,1当量)的甲醇(2.0毫升)溶液中加入盐酸的1,4-二氧六环溶液(4摩尔每升,2.0毫升)。所得混合物在25摄氏度下继续搅拌反应1.5小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩除去多余的试剂,所得混合物通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的乙腈/水流动相(0.1%盐酸)进行洗脱;检测器UV220/254纳米;得到化合物39(黄色固体,54.8毫克,产率77%)。MS(ESI,m/z):1099.1/1101.1[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ11.20–10.98(m,1H),10.14–9.93(m,1H),9.86–9.66(m,1H),9.19–9.09(m,1H),8.72–8.61(m,1H),8.55(d,J=9.1Hz,1H),8.04–7.97(m,1H),7.97–7.90(m,1H),7.81(d,J=8.3Hz,1H),7.50–7.36(m,5H),7.34–7.28(m,1H),7.28–7.17(m,2H),7.12(d,J=2.3Hz,1H),5.25–5.19(m,2H),4.79–4.70(m,2H),4.62–4.39(m,6H),4.37–4.30(m,1H),4.29–4.09(m,4H),4.07–3.91(m,3H),3.85–3.61(m,3H),3.60–3.53(m,1H),3.52–3.41(m,1H),3.31–3.10(m,3H),2.47(s,3H),2.33–2.20(m,1H),2.13–1.84(m,9H),0.96(s,9H);
19F NMR(282MHz,DMSO-d
6)δ-121.84。
实施例40
(2S,4R)-1-((S)-2-(3-(3-((S)-2-(((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基) -7-(8-乙基-7-氟-3-羟基萘-1-基)-6,8-二氟喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丙氧基)丙酰胺基)-3,3-二甲基丁酰)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二甲酸盐40
步骤1
用3-氯-2,4-二氟苯胺替代2-氯-3-氟-4-氨基吡啶,参考化合物38-6的合成方法,得到化合物40-1(淡黄色固体)。MS(ESI,m/z):489.1/491.0[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.44–7.36(m,1H),4.48–4.31(m,4H),3.75–3.54(m,2H),2.08–1.92(m,2H),1.79–1.70(m,2H),1.54(s,9H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向化合物40-1(2.5克,4.849毫摩尔,1当量),碳酸铯(4.99克,14.547毫摩尔,3当量)和三乙烯二胺(0.11克,0.970毫摩尔,0.2当量)的N,N-二甲基甲酰胺(25毫升)溶液中加入化合物25-4(1.31克,5.819毫摩尔,1.2当量)。混合物在90摄氏度条件下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,加入200毫升水稀释反应液,水层用乙酸乙酯(300毫升 x 3)萃取,合并后有机层用饱和食盐水(100毫升 x 3)洗涤,再用无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物40-2(黄色油,1.13克,产率29%)。MS(ESI,m/z):666.1/667.9[M+H]
+。
步骤3
在25摄氏度氮气保护搅拌条件下,向化合物40-2(800毫克,1.14毫摩尔,1.0当量),化合物40-3(518.78毫克,1.368毫摩尔,1.2当量,参照专利(WO2021041671(A1))合成得到),2-二环己基膦-2',4',6'-三异丙基联苯(114.42毫克,0.228毫摩尔,0.2当量)和三(二亚苄基丙酮)二钯(109.90毫克,0.114毫摩尔,0.1当量)的1,4-二氧六环/水(20毫升/5毫升)的混合溶液中分批加入碳酸铯(782.04毫克,2.280毫摩尔,2当量)。混合物在80摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,混合物浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→6%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到的粗产品再通过高效液相色谱进行纯化,流动相A:水(0.05%三氟乙酸),流动相B:甲醇;流速:100毫升/分钟;在30分钟内用50%→90%的B相进行洗脱,检测器UV 220/254纳米。得到化合物40-4(黄色油,320毫克,产率33%)。MS(ESI,m/z):820.2/822.2[M+H]
+。
步骤4
通过超临界液相色谱对步骤3所得化合物40-4(320毫克)进行手性拆分:手性柱CHIRAL ART Amylose-SA,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:异丙醇(0.5%,2摩尔/升氨甲醇);流速:60毫升/分钟;柱温:35摄氏度;用35%流动相B洗脱;检测器UV222纳米,得到两个产品。较短保留时间(5.15分钟)的产品为化合物40-4a(白色固体,123毫克,回收率38%),化合物40-4a:MS(ESI,m/z):820.2/822.2[M+H]
+;较短保留时间(6.32分钟)的产品为化合物40-4b(白色固体,106毫克,产率33%),化合物40-4b:MS(ESI,m/z):820.2/822.2[M+H]
+。
步骤5
在0摄氏度搅拌条件下,向反应瓶中依次加入化合物40-4a(100毫克,0.116毫摩尔,1当量),高碘酸钠(130.43毫克,0.580毫摩尔,5当量),四氯化碳(2.0毫升),乙腈(2毫升),向其中慢慢 加入三氯化钌(2.53毫克,0.012毫摩尔,0.1当量)的水(3毫升)溶液。混合物在0摄氏度条件下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%氨水,氨水中的氨含量25%~28%);流动相B:甲醇,在20分钟内用30%→95%的B相进行洗脱;检测器UV254/220纳米;得到化合物40-5a(黄色固体,60.0毫克,产率59%)。MS(ESI,m/z):838.5/840.5[M+H]
+。
步骤6
在室温搅拌条件下,向化合物40-5a(50.00毫克,0.057毫摩尔,1.00当量)的N,N-二甲基甲酰胺(1毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(27.23毫克,0.068毫摩尔,1.2当量)。混合物在25摄氏度下搅拌反应10分钟,再向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(30.65毫克,0.063毫摩尔,1.1当量)和N,N-二异丙基乙胺(29.31毫克,0.228毫摩尔,4当量)。混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,所得混合物通过高效液相色谱进行纯化,色谱柱XBridge Prep C18 OBD Column 30 x 50毫米,5微米;流动相A:水(0.05%碳酸氢铵),流动相B:乙腈;流速:60毫升/分钟;在8分钟内用55%→80%的B相进行洗脱,检测器UV 220纳米。得到化合物40-6a(白色固体,30毫克,产率40%)。MS(ESI,m/z):1250.0/1252.1[M+H]
+。
步骤7
在0摄氏度搅拌条件下,向化合物40-6a(30.00毫克,0.114毫摩尔,1.00当量)的甲醇(2毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,2毫升)。混合物在室温下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过高效液相色谱进行纯化,色谱柱XBridge Shield RP18 OBD Column,19 x 150毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;在1.5分钟内用5%→5%的B,在2分钟内用5%→13%的B,在9分钟内用13%→39%的B相进行洗脱,检测器UV 220纳米。得到化合物40(白色固体,14.2毫克,产率51%)。MS(ESI,m/z):1106.1/1108.1[M+H]
+。
1H NMR(300MHz,CD
3OD)δ8.89(s,1H),8.73–8.34(m,2H),7.76–7.61(m,2H),7.52–7.37(m,4H),7.34–7.19(m,2H),7.00(d,J=2.6Hz, 1H),4.79–4.62(m,3H),4.61–4.44(m,5H),4.44–4.33(m,1H),4.02–3.51(m,14H),3.25–3.01(m,2H),2.66–2.39(m,7H),2.37–2.16(m,2H),2.16–1.90(m,10H),1.09–0.95(m,9H),0.89–0.75(m,3H);
19F NMR(282MHz,CD
3OD)δ-117.19,-120.99,-124.04。
实施例41
(2S,4R)-1-((S)-2-(4-((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)丙基)(甲基)氨基)丁酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺二盐酸盐41
步骤1
在0摄氏度氮气保护条件下,向化合物16-3a(300毫克,0.465毫摩尔,1.00当量)和化合物4-1(93.81毫克,0.567毫摩尔,1.2当量)的无水四氢呋喃(3毫升)溶液中缓慢滴加叔丁醇钾的四氢呋喃溶液(1摩尔每升,0.56毫升,0.56毫摩尔,1.2当量)。所得混合物在0摄氏度条件下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,向体系中加入10毫升水淬灭反应,水相用乙酸乙酯萃取(10毫升 x 3),合并后有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%的甲醇/二氯甲烷的流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物41-1(黄色固体,231毫克,产率64%)。MS(ESI,m/z):734.5/736.5[M+H]
+。
步骤2
在0摄氏度搅拌条件下,向反应瓶中依次加入41-1(160毫克,0.207毫摩尔,1.0当量),高碘酸钠(186.42毫克,0.828毫摩尔,4.0当量),四氯化碳(1.0毫升),乙腈(1毫升)。然后向其中慢 慢加入三氯化钌(2.25毫克,0.009毫摩尔,0.05当量)的水(1.5毫升)溶液。混合物在0摄氏度条件下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,向体系中加入15毫升水稀释反应液,再用氯仿/异丙醇(3/1,20毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→95%的乙腈/水流动相(0.1%碳酸氢铵)进行洗脱;检测器UV254纳米;得到化合物41-2(白色固体,70毫克,产率40%)。MS(ESI,m/z):752.4/754.4[M+H]
+。
步骤3
在25摄氏度搅拌条件下,向反应瓶中依次加入41-2(70毫克,0.088毫摩尔,1.0当量),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(53.07毫克,0.132毫摩尔,1.5当量)和N,N-二甲基甲酰胺(2毫升)。混合物在25摄氏度条件下反应15分钟。然后向上述反应体系中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(41.37毫克,0.088毫摩尔,1当量)和N,N-二异丙基乙胺(48.1毫克,0.354毫摩尔,4当量)。混合物在25摄氏度条件下继续反应2小时。反应过程通过液质和薄层层析来监控。反应完全后,粗产品通过反相快速色谱柱(C18柱)进行纯化,在10分钟内用10%→95%的甲醇/水流动相进行洗脱;检测器UV220纳米;得到化合物41-3(黄色固体,54毫克,产率49%)。MS(ESI,m/z):1164.7/1166.7[M+H]
+。
步骤4
在0摄氏度搅拌条件下,向化合物41-3(54毫克,0.044毫摩尔,1.00当量)的甲醇(2.0毫升)溶液中加入盐酸的1,4-二氧六环溶液(4摩尔每升,2.0毫升)。所得混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品,所得粗产品通过反相快速色谱柱(C18柱)进行纯化,在30分钟内用5%→95%的甲醇/水流动相(0.1%盐酸)进行洗脱;检测器UV220/254纳米;得到化合物41(黄色固体,22.5毫克,产率45%)。MS(ESI,m/z):1020.5/1022.5[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ10.71–10.52(m,1H),10.10–9.96(m,1H),9.88–9.68(m,1H),9.12(s,1H),8.72–8.60(m,1H),8.19–8.09(m,1H),8.06(d,J=1.6Hz,1H),7.88(d,J=8.3Hz,1H),7.55–7.41(m,5H),7.38(d,J=2.4Hz,1H),7.34–7.23(m,2H),7.17(d,J=2.4Hz,1H),4.67–4.38(m,8H),4.35–4.17(m,3H),4.09–3.93(m,2H),3.78–3.65(m,2H),3.40–3.02(m,4H),2.83(d,J =4.7Hz,3H),2.52(s,3H),2.44–2.33(m,2H),2.32–2.21(m,2H),2.17–1.90(m,8H),1.01(s,9H);
19F NMR(282MHz,DMSO-d
6)δ-121.99。
实施例42
(2S,4R)-1-((S)-2-((1R,4R或1S,4S)-4-(2-((S)-2-(((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙基)环己烷-1-甲酰胺基-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺甲酸盐42a;(2S,4R)-1-((S)-2-((1S,4S或1R,4R)-4-(2-((S)-2-(((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)乙基)环己烷-1-甲酰胺基-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺甲酸盐42b
步骤1
在0摄氏度搅拌条件下,向2-(4-(乙氧羰基)环己基)乙酸(1.0克,4.4毫摩尔,1.0当量)的无水四氢呋喃(1毫升)溶液中缓慢滴加硼烷的四氢呋喃溶液(1摩尔/升,6.6毫升,6.6毫摩尔,1.0当量)。混合物在25摄氏度下搅拌2小时,反应过程通过液质和薄层层析来监控。反应完成后,将反应液缓慢倒入冰水中,再用乙酸乙酯(30毫升 x 3)萃取,合并后有机相用无水硫酸钠干燥。过滤除去不溶物,有机相减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用0%→9%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物42-1(黄色油,698毫克,产率74%)。MS(ESI,m/z):201.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ4.19–4.06(m,2H),3.69(t,J=6.6Hz,2H),2.28–2.15(m,1H),2.03–1.92(m,2H),1.88–1.78(m,2H),1.64–1.35(m,5H),1.34–1.21(m,3H),1.05–0.90(m,2H)。
步骤2
在0摄氏度氮气保护搅拌条件下,向化合物42-1(690毫克,3.2毫摩尔,1.0当量),N,N-二甲氨基吡啶(42毫克,0.3毫摩尔,0.1当量)和三乙胺(1.44毫升,9.8毫摩尔,3.0当量)的二氯甲烷(6毫升)溶液中缓慢加入对甲苯磺酰氯(788毫克,毫摩尔,1.2当量)。所得混合物在20摄氏度下搅拌反应2小时。反应过程通过液质和薄层层析来监控。反应结束后,将反应液缓慢倒入冰水中,再用二氯甲烷(20毫升 x 3)萃取,合并后有机相用无水硫酸钠干燥。过滤除去干燥剂,有机相减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用0%→50%甲基叔丁基醚/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物42-2(淡黄色油,1.1克,产率80%)。MS(ESI,m/z):355.1[M+H]
+。
1H NMR(400MHz,CDCl
3)δ7.79(d,J=8.1Hz,2H),7.35(d,J=8.0Hz,2H),4.15–4.02(m,4H),2.45(s,3H),2.20–2.10(m,1H),1.96–1.89(m,2H),1.73–1.64(m,2H),1.57–1.51(m,2H),1.41–1.28(m,3H),1.27–1.21(m,3H),0.95–0.82(m,2H)。
步骤3
在25摄氏度搅拌条件下,向反应瓶中依次加入化合物42-2(979毫克,2.6毫摩尔,1.0当量),L-脯氨醇(307毫克,2.8毫摩尔,1.1当量),碳酸钾(763毫克,5.2毫摩尔,2当量)和乙腈(10毫升)。所得混合物在60摄氏度下反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,过滤除去不溶物,滤液减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用0%→9%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物42-3(淡黄色油,600毫克,产率76%)。MS(ESI,m/z):284.1[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.17–4.08(m,2H),3.80–3.69(m,1H),3.63–3.50(m,1H),3.47–3.31(m,1H),3.03–2.78(m,1H),2.57–2.36(m,2H),2.28–2.16(m,1H),2.06–1.73(m,9H),1.66–1.23(m,8H),1.09–0.89(m,2H)。
步骤4
在-20摄氏度氮气保护搅拌条件下,向化合物16-3a(1.18克,1.92毫摩尔,1.0当量)和化合物 42-3(600毫克,2.12毫摩尔,1.1当量)的无水四氢呋喃(10毫升)溶液中滴加叔丁醇钾的四氢呋喃溶液(1摩尔/升,2.88毫升,2.88毫摩尔,1.5当量)。所得混合物在-20摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,将反应液缓慢倒入冰水中,再用乙酸乙酯(30毫升 x 3)萃取,合并后有机相用无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得粗产品。粗产品用硅胶柱层析法纯化,用0%→9%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物42-4(白色固体,791毫克,产率48%)。MS(ESI,m/z):860.2/862.2[M+H]
+。
步骤5
通过制备级手性高效液相色谱法对步骤4所得化合物42-4(576毫克)进行手性拆分:手性柱Lux5m Cellulose-2,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:(乙醇:乙腈=1:2);流速:70毫升/分钟;在25分钟内用35%的B相进行洗脱,检测器UV 225纳米,得到两个产品。较短保留时间(8.68分钟)的产品为化合物42-4a(白色固体,49毫克,回收率8%),化合物42-4a:MS(ESI,m/z):860.2/862.2[M+H]
+;较长保留时间(11.33分钟)的产品为化合物42-4b(白色固体,268毫克,回收率46%),化合物42-4b:MS(ESI,m/z):860.2/862.2[M+H]
+。
步骤6
在25摄氏度搅拌条件下,向化合物42-4b(130毫克,19.0毫摩尔,1.0当量)的水/甲醇/四氢呋喃(2/1/2,5毫升)的混合溶液中加入氢氧化锂(36毫克,1.4毫摩尔,10.0当量)。混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩除去多余的试剂。向体系中加入6毫升水稀释混合物,再滴加1摩尔每升的稀盐酸调节pH至5。水相用二氯甲烷(10毫升 x 3)萃取,合并的有机相用无水硫酸钠干燥,过滤除去干燥剂;滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→9%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压浓缩除去溶剂,得到化合物42-5b(白色固体,51毫克,产率40%)。MS(ESI,m/z):832.2/834.2[M+H]
+。
步骤7
在25摄氏度搅拌条件下,向化合物42-5b(50毫克,0.05毫摩尔,1.0当量)的N,N-二甲基甲酰胺(1.00毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(27毫克,0.06毫摩尔,1.2当量)。混合物在25摄氏度搅拌反应30分钟。然后向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(33毫克,0.06毫摩尔,1.2当量)和N,N-二异丙基乙胺(41微升,0.17毫摩尔,4当量)。混合物在25摄氏度下继续搅拌反应1小时。反应过程通过液质和薄层层析来监控。反应完全后,混合物通过反相快速色谱柱(C18柱)进行纯化,在25分钟内用5%→70%的乙腈/水流动相(0.05%三氟乙酸)进行洗脱;检测器UV254纳米;得到化合物42-6b(白色固体,84毫克,产率43%)。MS(ESI,m/z):1244.2/1246.2[M+H]
+。
步骤8
在0摄氏度搅拌条件下,向化合物42-6b(80.0毫克,0.06毫摩尔,1当量)的甲醇(2.0毫升)溶液中加入盐酸的1,4-二氧六环溶液(4摩尔/升,2.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩除去多余的试剂,所得混合物通过制备级高效液相色谱进行纯化,色谱柱Xselect CSH F-Phenyl OBD,19 x 250毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:25毫升/分钟;在8分钟内用15%→35%的B相进行洗脱,检测器UV 254/220纳米。得到化合物42b(白色固体,15毫克,产率21%)。MS(ESI,m/z):1100.1/1102.1[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ10.08(s,1H),9.50(s,1H),8.99(s,1H),8.62–8.51(m,1H),8.04–7.95(m,1H),7.82(d,J=8.3Hz,1H),7.66(d,J=9.2Hz,1H),7.49–7.36(m, 5H),7.31(d,J=2.4Hz,1H),7.25–7.16(m,2H),7.08(d,J=2.4Hz,1H),5.23–5.08(m,1H),4.80–4.39(m,7H),4.39–4.18(m,3H),4.18–4.09(m,2H),3.94–3.79(m,2H),3.72–3.58(m,2H),3.12–2.87(m,1H),2.55(s,1H),2.45(s,3H),2.36–2.23(m,1H),2.22–2.10(m,1H),2.09–1.78(m,9H),1.77–1.62(m,4H),1.62–1.56(m,1H),1.56–1.42(m,2H),1.39–1.13(m,4H),1.05–0.70(m,11H);
19F NMR(282MHz,DMSO-d
6)δ-121.90。
按照步骤6,步骤7,步骤8中描述的方法,从化合物42-4a出发得到化合物42a(白色固体,2.3毫克),MS(ESI,m/z):1100.1/1102.1[M+H]
+。
实施例43
4-((3-(3-((S)-2-(((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛基-3-基)-6-氯-8-氟-7-(3-羟基萘-1-基)喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丙氧基)丙基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮三三氟乙酸盐43
步骤1
在0摄氏度搅拌条件下,向N-(3-羟丙基)氨基甲酸叔丁酯(10克,54.215毫摩尔,1.00当量)和N,N-二异丙基乙胺(11.80克,86.744毫摩尔,1.6当量)的二氯甲烷(100.0毫升)溶液中缓慢加入甲基磺酰氯(8.50克,70.480毫摩尔,1.3当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过薄层层析来监控。反应完全后,将反应液倒入冰水中淬灭反应,水相用二氯甲烷(200毫升 x 3)萃取。合并后有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→50%乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物43-1(黄色油,14克,产率96%)。
1H NMR(300MHz,CDCl
3)δ4.33–4.29(m,2H),3.30–3.26(m,2H),3.05(s,3H),2.00–1.91(m,2H),1.46(s,9H)。
步骤2:
在0摄氏度氮气保护搅拌条件下,向1,3-丙二醇(26.64克,332.639毫摩尔,6.57当量)的N,N-二甲基甲酰胺(253毫升)溶液中加入氢化钠(60%,4.05克,101.260毫摩尔,2.00当量)。混合物在0摄氏度下搅拌0.5小时后,加入化合物43-1(13.5克,50.630毫摩尔,1.00当量)。反应液在25摄氏度条件下反应16小时,反应过程通过薄层层析来监控。反应完全后,反应液用饱和食盐水清洗(1000毫升 x 3),有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物43-2(黄色油,4.5克,产率36%)。
1H NMR(400MHz,CDCl
3)δ3.78(t,J=5.8Hz,2H)3.59(t,J=5.8Hz,2H),3.50(t,J=5.9Hz,2H),3.23(t,J=6.5Hz,2H),1.86–1.79(m,2H),1.78–1.70(m,2H),1.44(s,9H)。
步骤3:
在25摄氏度搅拌条件下,向化合物43-2(400毫克,1.629毫摩尔,1.00当量)的甲醇(2毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,2毫升)。混合物在25摄氏度搅拌条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品43-3(520毫克)。粗产品直接用于下一步反应。MS(ESI,m/z):134.2[M+H]
+。
步骤4:
在25摄氏度,氮气保护搅拌条件下,向化合物43-3(297.99毫克,2.125毫摩尔,1.20当量),N-甲基吡咯烷酮(5.0毫升)和2-(2,6氧代-哌啶-3-基)-4-氟基-异吲哚-1,3-二酮(515毫克,1.771毫摩尔,1.00当量)的混合物中滴加N,N-二异丙基乙胺(722.90毫克,5.313毫摩尔,3.00当量)。将所得 混合物在90摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,将反应液冷却至25摄氏度。混合物通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用5%→60%的乙腈/水流动相进行洗脱;检测器UV254纳米;得到化合物的粗产品。粗产品用硅胶柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物43-4(黄色固体,300毫克,产率41%)。MS(ESI,m/z):390.1[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ11.10(s,1H),7.64–7.49(m,1H),7.17–6.91(m,2H),6.80–6.54(m,1H),5.76(s,1H),5.15–4.93(m,1H),4.38(s,1H),3.49–3.34(m,8H),2.96–2.79(m,1H),2.64–2.57(m,1H),2.11–1.96(m,1H),1.86–1.74(m,2H),1.73–1.59(m,2H)。
步骤5:
在0摄氏度氮气保护搅拌条件下,向化合物43-4(300毫克,0.732毫摩尔,1.00当量)的二氯甲烷(6毫升)溶液中加入1,1,1-三乙酰氧基-1,1-二氢-1,2-苯碘酰-3(1H)-酮(392.11毫克,0.878毫摩尔,1.20当量)。所得混合物在25摄氏度下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物43-5(黄色固体,130毫克,产率43%)。MS(ESI,m/z):388.2[M+H]
+。
步骤6
在0摄氏度搅拌条件下,向化合物35-2(240毫克,0.33毫摩尔,1当量)的甲醇(3.0毫升)溶液中加入盐酸的1,4-二氧六环溶液(4摩尔/升,3.0毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,减压浓缩得到粗产品43-6(310毫克)。MS(ESI,m/z):574.2/576.2[M+H]
+。
步骤7
在0摄氏度搅拌条件下,向化合物43-6(300毫克,0.318毫摩尔,1.00当量)和N,N-二异丙基乙胺(151.28毫克,1.113毫摩尔,3.50当量)的二氯甲烷(6.0毫升)溶液中加入二碳酸二叔丁酯(72.99毫克,0.318毫摩尔,1.00当量)。所得混合物在0摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品。粗产品用硅胶柱层析法纯化,用0%→6%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物43-7(黄色固体,210毫克)。MS(ESI,m/z):674.3/676.3[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ10.02(s,1H),8.00(d,J=1.6Hz,1H),7.81(d,J=8.3Hz,1H),7.49–7.37(m,1H),7.29(d,J=2.4Hz,1H),7.25–7.18(m,2H),7.07(d,J=2.4Hz,1H),5.93–5.79(m,1H),5.16(d,J=16.9Hz,1H),5.03(d,J=10.3Hz,1H),4.50–4.33(m,3H),4.31–4.20(m,2H),4.21–4.09(m,1H),3.65–3.49(m,3H),3.04–2.91(m,2H),2.90–2.79(m,1H),2.31–2.16(m,1H),1.98–1.59(m,8H),1.47(s,9H)。
步骤8
在25摄氏度氮气保护条件下,依次向三口瓶中加入化合物43-7(200毫克,0.282毫摩尔,1.00当量),1,3-二甲基巴比妥酸(69.48毫克,0.423毫摩尔,1.50当量),四(三苯基膦)钯(17.14毫克,0.014毫摩尔,0.05当量)和二氯甲烷(6毫升)。所得混合物在25摄氏度反应5小时,反应过程通过液质和薄色谱层层析监控。反应结束后,减压浓缩除去多余的溶剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用20%→60%乙腈/水流动相(0.1%甲酸)进行洗脱,检测器UV254纳米,得到化合物43-8(淡黄色固体,120毫克,产率63%)。MS(ESI,m/z):634.2/636.2[M+H]
+。
步骤9
在25摄氏度搅拌条件下,向化合物43-8(120毫克,0.180毫摩尔,1.00当量),化合物43-5(109.96毫克,0.270毫摩尔,1.50当量)和醋酸(22.73毫克,0.360毫摩尔,2.00当量)的甲醇(7毫升)溶液中加入氰基硼氢化钠(17.84毫克,0.270毫摩尔,1.50当量)。所得混合物在25摄氏度反应1小时,反应过程通过液质和薄色谱层层析监控。反应结束后,反应液减压浓缩得到粗产品。所得粗产品用硅胶柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物43-9(黄色固体,75毫克,产率37%)。MS(ESI,m/z):1005.4/1007.4[M+H]
+。
步骤10
在室温搅拌条件下,向化合物43-9(70毫克,0.069毫摩尔,1.00当量)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1毫升)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液减压浓缩得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在10分钟内用5%→50%的乙腈/水流动相(0.1%三氟乙酸)进行洗脱,检测器UV254纳米,得到化合物43(黄绿色固体,40毫克,产率43%)。MS(ESI,m/z):905.5/907.5[M+H]
+;
1H NMR(300MHz,DMSO-d
6)δ7.92(d,J=14.9Hz,1H),7.81(d,J=8.3Hz,1H),7.53–7.42(m,2H),7.31(d,J=2.4Hz,1H),7.27–7.13(m,2H),7.10–7.06(m,1H),6.98–6.91(m,2H),5.10–4.98(m,1H),4.70–4.40(m,4H),4.25–4.12(m,2H),4.00–3.90(m,1H),3.88–3.71(m,2H),3.69–3.56(m,2H),3.47–3.37(m,4H),3.32–3.15(m,4H),2.94–2.78(m,1H),2.67–2.54(m,2H),2.33–2.20(m,1H),2.08–1.88(m,10H),1.81–1.68(m,2H);
19F NMR(282MHz,DMSO-d
6)δ-74.13,-121.86,-121.92。
实施例44
(2S,4R)-1-((2S)-2-(3-(3-((2R,7aS)-7a-((4-(3,8-二氮杂双环[3.2.1]辛烷-3-基)-7-(8-乙炔基-7-氟-3-羟基萘-1-基)-8-氟吡啶[4,3-d]嘧啶-2-基)氧基)甲基)六氢-1H-吡咯嗪-2-基)氧基)丙氧基)丙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苄基)吡咯烷-2-甲酰胺三氟乙酸盐(44)
步骤1:
在0摄氏度氮气保护搅拌条件下,向化合物2,5-二氧代-1H-四氢吡咯嗪-7a(5H)-羧酸乙酯(10.5克,47.226毫摩尔,1当量)的无水四氢呋喃(150毫升)溶液中分批加入氢化铝锂(9.43克,236.13毫摩尔,5当量)。混合物在60摄氏度搅拌条件下反应16小时,反应过程通过液质和薄层层析来监控。反应结束后,将混合物体系冷却至室温,依次加入水(9.5毫升)、15%氢氧化钠水溶液(9.5毫升)和水(28.5毫升)。过滤除去不溶物,滤液减压浓缩得到粗产品。所得粗产品通过高效液相色谱进行纯化,色谱柱XBridge Prep Phenyl OBD Column,19 x 150毫米,5微米;流动相A:水(0.05%氨水),流动相B:甲醇;流速:25毫升/分钟;在7分钟内用3%→30%的B相进行洗脱,检测器UV200/220纳米。得到化合物44-1(淡黄色油状液体,2.66克,产率34%)。MS(ESI,m/z):158.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ4.54(s,2H),4.47–4.35(m,1H),3.30–3.13(m,3H),2.99–2.78(m,2H),2.61–2.52(m,1H),2.25–2.16(m,1H),2.04–1.86(m,1H),1.83–1.55(m,4H)。
步骤2:
在-40摄氏度氮气保护搅拌条件下,向化合物44-1(2.6克,15.711毫摩尔,1当量)的N,N-二甲基甲酰胺(30毫升)溶液中分批加入叔丁基二苯基氯硅烷(4.91克,16.97毫摩尔,1.08当量)。混合物在25摄氏度搅拌条件下反应5小时,反应过程通过液质和薄层层析来监控。反应结束后,混合物体系通过反相色谱柱(C18柱)进行纯化得到粗产品,在25分钟内用5%→95%的甲醇/水流动相(0.1%甲酸)进行洗脱;检测器,UV254/220纳米。粗品通过制备级超临界液相色谱法进一步纯化,纯化条件:手性柱YMC-PACK CN,3 x 25厘米,5微米;流动相A:超临界二氧化碳流体,流动相B: 甲醇(0.1%2摩尔/升氨甲醇);流速:180毫升/分钟;在12分钟内用30%的B相进行洗脱,检测器UV 220/204纳米,得到化合物44-2(淡黄色油状液体,2.4克,产率36%)。MS(ESI,m/z):396.1[M+H]
+;
1H NMR(400MHz,CDCl
3)δ9.14–9.06(m,4H),8.88–8.77(m,6H),5.89–5.80(m,1H),4.83–4.75(m,2H),4.72–4.65(m,1H),4.52–4.43(m,1H),4.30–4.21(m,1H),4.09–4.00(m,1H),3.80–3.72(m,1H),3.40–3.30(m,2H),3.21–3.13(m,2H),3.08–3.02(m,1H),2.50(s,9H)。
步骤3:
通过制备级超临界液相色谱法对步骤2所得化合物44-2(2.4克)进行手性拆分:手性柱Lux 5 m Cellulose-4,3 x 25厘米,5微米;流动相A:超临界二氧化碳流体,流动相B:甲醇(0.1%2摩尔/升氨甲醇);流速:20毫升/分钟;在12分钟内用35%的B相进行洗脱,检测器:UV 204纳米,得到两个产品。较短保留时间(5.83分钟)的产品为化合物44-3(黄色油状物,1.05克,回收率44%),MS(ESI,m/z):396.1[M+H]
+;[α]
25
D=+11.6(c=0.33g/100mL,MeOH)。
步骤4:
在0摄氏度氮气保护搅拌条件下,向化合物44-3(800毫克,1.921毫摩尔,1.0当量)和碘化钠(333.42毫克,2.113毫摩尔,1.1当量)的无水N,N-二甲基甲酰胺(8毫升)溶液中滴加1摩尔/升叔丁醇钾四氢呋喃溶液(3.84毫升,3.84毫摩尔,2.0当量)。混合物在25摄氏度搅拌条件下反应16小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液通过反相色谱柱(C18柱)进行纯化得到粗产品,在25分钟内用10%→85%的甲醇/水流动相(0.1%甲酸)进行洗脱;检测器:UV254/220纳米。所得粗产品通过高效液相色谱进行进一步纯化,色谱柱YMC-Actus Triart C18 ExRS,30 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:甲醇;流速:60毫升/分钟;在10分钟内用85%→95%的B相进行洗脱,检测器:UV 220/254纳米。得到化合物44-4(黄色油状液体,380毫克,产率37%)。MS(ESI,m/z):508.3[M+H]
+。
步骤5:
在25摄氏度搅拌条件下,向化合物44-4(350毫克,0.655毫摩尔,1.0当量)的四氢呋喃(4毫升)溶液中滴加1摩尔/升四丁基氟化铵四氢呋喃溶液(1.64毫升,1.64毫摩尔,2.5当量)。混合物在40摄氏度搅拌条件下反应18小时,反应过程通过液质和薄层层析来监控。反应结束后,减压浓缩得到粗产品。粗产品通过硅胶柱层析进行纯化,流动相用0%→10%甲醇(1%,7摩尔/升氨甲醇)/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂得到化合物44-5(黄色油状液体,140毫克,产率78%)。MS(ESI,m/z):270.2[M+H]
+,
1H NMR(400MHz,CDCl
3)δ5.87–5.77(m,1H),5.15–5.00(m,2H),4.05–4.00(m,1H),3.50–3.43(m,6H),3.25(s,2H),3.16–3.12(m,1H),3.10–3.04(m,1H),2.90–2.85(m,1H),2.82–2.78(m,1H),2.35–2.30(m,2H),2.10–2.05(m,1H),1.99–1.88(m,1H),1.84–1.72(m,6H)。
步骤6:
在-10摄氏度氮气保护搅拌条件下,向化合物44-5(110毫克,0.388毫摩尔,1.0当量)和化合物38-6(210毫克,0.166毫摩尔,1.2当量)的四氢呋喃(3毫升)溶液中滴加1摩尔/升叔丁醇钾四氢呋喃溶液(0.58毫升,0.58毫摩尔,1.5当量)。混合物在-10摄氏度搅拌条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后向反应液中加入水(25毫升)淬灭反应,所得混合物用乙酸乙酯(25毫升 x 3)萃取,合并有机相;有机相用饱和食盐水(30毫升 x 1)清洗,再用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→8%甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂得到化合物44-6(黄色固体,190毫克,产率68%)。MS(ESI,m/z):661.2/663.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ8.73(s,1H),5.86–5.76(m,1H),5.15–5.01(m,2H),4.65–4.29(m,6H),4.20(s,1H),3.65(d,J=7.5Hz,2H),3.55–3.43(m,7H),3.05(s,2H),2.35–2.29(m,3H),2.18(s,1H),2.11–1.91(m,6H),1.86–1.80(m,2H),1.78–1.65(m,3H),1.52(s,9H)。
步骤7:
在25摄氏度氮气保护搅拌条件下,依次向反应瓶中加入化合物44-6(110毫克,0.158毫摩尔,1.0当量),((2-氟-6-(甲氧基甲氧基)-8-(4,4,5,5-四甲基-1,3,2-二氧苯甲醛-2-基)萘-1-基)乙炔基)三异丙基硅烷(参考WO2021041671制得,110.8毫克,0.205毫摩尔,1.3当量),三(二亚苄基丙酮)二钯(15.2毫克,0.016毫摩尔,0.1当量),正丁基二(1-金刚烷基)膦(11.9毫克,0.032毫摩尔,0.2当量),甲苯(2毫升)和水(0.4毫升)。混合物在80摄氏度搅拌条件下反应8小时,反应过程通过液质和薄层层析来监控。反应结束后冷却至室温,向反应液中加入水(10毫升)稀释,所得混合物用乙酸乙酯(10毫升 x 3)萃取,合并有机相;有机相用30毫升饱和食盐水洗涤,再用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。所得粗产品通过硅胶柱层析进行纯化,流动相用0%→8%甲醇/二氯甲烷梯度洗脱,所得馏分通过减压旋蒸除去溶剂得到化合物44-7(黄色固体,120毫克,产率71%)。MS(ESI,m/z):1011.5[M+H]
+;
1H NMR(400MHz,CDCl
3)δ9.06(s,1H),7.80–7.76(m,1H),7.51(d,J=2.6Hz,1H),7.34–7.28(m,2H),5.8–5.76(m,1H),5.30(s,2H),5.14–5.02(m,2H),4.83–4.68(m,2H),4.54–3.99(m,6H),3.96–3.71(m,2H),3.61–3.45(m,11H),2.51–2.12(m,7H),2.10–1.93(m,5H),1.90–1.80(m,3H),1.52(s,9H),0.88–0.84(m,18H),0.63–0.46(m,3H)。
步骤8:
在0摄氏度搅拌条件下,向化合物44-7(110毫克,0.103毫摩尔,1.0当量),一水合三氯化钌(2.5毫克,0.01毫摩尔,0.1当量),水(3毫升),四氯化碳(2毫升)和乙腈(2毫升)的混合溶液中分批加入高碘酸钠(116.3毫克,0.515毫摩尔,5.0当量)。混合物在0摄氏度搅拌条件下反应1小时,反应过程通过液质监控。反应结束后,向反应液中加入水(10毫升),所得混合物用氯仿/异丙醇(3/1,10毫升 x 3)萃取,合并有机相,无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过反相色谱柱(C18柱)进行纯化,在15分钟内用5%→95%的甲醇/水流动相(0.1%氨水)进行洗脱;检测器:UV254/220纳米;得到化合物44-8(白色固体,70毫克,产率62%)。MS(ESI,m/z):1029.5[M+H]
+。
步骤9:
在25摄氏度搅拌条件下,向化合物44-8(70毫克,0.065毫摩尔,1.0当量)的N,N-二甲基甲酰 胺(2毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(31.0毫克,0.078毫摩尔,1.2当量)。混合物在25摄氏度下反应15分钟。然后向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(38.1毫克,0.078毫摩尔,1.2当量)和N,N-二异丙基乙胺(35.1毫克,0.26毫摩尔,4当量)。反应液在25摄氏度下反应1小时,反应过程通过液质和来监控。反应结束后,混合物通过反相色谱柱(C18柱)进行纯化,在15分钟内用5%→95%的甲醇/水流动相(0.1%三氟乙酸)进行洗脱;检测器:UV254/220纳米;得到化合物44-9(黄色固体,68毫克,产率69%)。MS(ESI,m/z):721.7[M/2+H]
+。
步骤10:
在0摄氏度搅拌条件下,向化合物44-9(68毫克,0.047毫摩尔,1.0当量)的乙腈(2.5毫升)溶液中加入4摩尔每升的盐酸1,4-二氧六环溶液(0.5毫升)。混合物在0摄氏度搅拌条件下反应2小时,反应过程通过液质监控。反应结束后,减压浓缩除去溶剂。向混合物中加入饱和碳酸氢钠溶液(10毫升),所得混合物用氯仿/异丙醇(3/1,10毫升 x 3)萃取,合并有机相,再用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品化合物44-10(黄色固体,60毫克,产率95%)。MS(ESI,m/z):1297.6[M+H]
+。
步骤11:
在25摄氏度搅拌条件下,向化合物44-10(60毫克,0.046毫摩尔,1.0当量)的N,N-二甲基甲酰胺(2毫升)溶液中加入氟化铯(76毫克,0.46毫摩尔,10当量)。混合物在25摄氏度搅拌条件下反应4小时,反应过程通过液质监控。反应结束后,过滤除去多余氟化铯,滤液减压浓缩得到粗产品。所得粗产品通过高效液相色谱进行纯化,色谱柱XBridge Prep OBD C18 Column,30 x 150毫米,5微米;流动相A:水(10毫摩尔/升碳酸氢铵),流动相B:乙腈;流速:60毫升/分钟;在7分钟内用27%→50%的B相进行洗脱,检测器UV 220/254纳米。得到化合物44(黄色固体,30毫克,产率49%)。MS(ESI,m/z):571.9[M/2+H]
+,1141.6[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ10.20(s,1H),9.08(s,1H),8.98(s,1H),8.62–8.59(m,1H),8.00–7.90(m,2H),7.54–7.35(m,6H),7.17(d,J=2.5Hz,1H),5.22–5.05(m,1H),4.62–4.52(m,2H),4.46–4.34(m,4H),4.29–4.01(m,4H),3.97–3.85(m,3H), 3.78–3.47(m,6H),3.44–3.37(m,4H),3.21–2.99(m,2H),2.92–2.69(m,2H),2.58–2.53(m,1H),2.43(s,3H),2.38–2.26(m,1H),2.19–1.99(m,2H),1.98–1.61(m,12H),0.92(s,9H);
19F NMR(377MHz,DMSO-d
6)δ-73.43,-110.72,-139.93。
实施例45
(2S,4R)-1-((S)-2-(3-(3-((S)-2-((((R或S)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-7-(8-乙基-7-氟-3-羟基萘-1-基)-6,8-二氟喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丙氧基)丙氨基)-3,3-二甲基丁酰-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺一甲酸盐45
步骤1
在室温搅拌条件下,向化合物40-5a(65.00毫克,0.074毫摩尔,1.00当量)的N,N-二甲基甲酰胺(1.5毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(35.39毫克,0.089毫摩尔,1.2当量)。混合物在25摄氏度下搅拌反应10分钟,再向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(41.38毫克,0.089毫摩尔,1.2当量)和N,N-二异丙基乙胺(40.10毫克,0.296毫摩尔,4当量)。混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,所得混合物通过高效液相色谱进行纯化,色谱柱Kinetex EVO C18 Column 21.2 x 150毫米,5微米;流动相A:水(0.05%碳酸氢铵),流动相B:乙腈;流速:25毫升/分钟;在8分钟内用80%→95%的B相进行洗脱,检测器UV 220纳米。得到化合物45-1(白色固体,42毫克,产率42%)。MS(ESI,m/z):1264.6[M+H]
+。
步骤2
在0摄氏度搅拌条件下,向化合物45-1(42.00毫克,0.032毫摩尔,1.00当量)的甲醇(1毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,1毫升)。混合物在室温下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过高效液相色谱进行纯化,色谱柱Sunfire prep C18 Column,30 x 150毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:60毫升/分钟;在7分钟内用18%→35%的B相进行洗脱,检测器UV 220纳米。得到化合物45(白色固体,25.8毫克,产率67%)。MS(ESI,m/z):1120.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ8.98(s,1H),8.38(d,J=7.8Hz,1H),8.29(s,1H),7.89–7.74(m,2H),7.73–7.63(m,1H),7.49–7.29(m,6H),7.02(d,J=2.6Hz,1H),5.01–4.81(m,1H),4.55–4.47(m,1H),4.47–4.37(m,1H),4.37–4.19(m,4H),4.12–4.02(m,1H),3.69–3.48(m,10H),3.43–3.30(m,3H),3.09–2.98(m,1H),2.93–2.76(m,2H),2.45(s,3H),2.40–2.15(m,4H),2.07–1.96(m,1H),1.95–1.86(m,1H),1.83–1.59(m,10H),1.36(d,J=7.0Hz,3H),0.89(s,9H),0.79–0.65(m,3H);
19F NMR(377MHz,DMSO-d
6)δ-118.286,-118.294,-119.111,-123.736,-123.742。
实施例46
4-((3-(3-((S)-2-((((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-7-(8-乙基-7-氟-3-羟基萘-1-基)-6,8-二氟喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丙氧基)丙基)氨基)-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮二盐酸盐46
步骤1
在25摄氏度氮气保护搅拌条件下,向化合物40-1(650毫克,1.261毫摩尔,1.0当量),化合物40-3(525.91毫克,1.387毫摩尔,1.1当量),3-(叔丁基)-4-(2,6-二甲氧基苯基)-2,3-二氢苯并[D][1,3]氧,膦戊轭(87.69毫克,0.252毫摩尔,0.2当量)和三(二亚苄基丙酮)二钯(121.54毫克,0.126毫摩尔,0.1当量)的甲苯/水(5.5毫升/1.1毫升)的混合溶液中分批加入磷酸钾(563.44毫克,2.522毫摩尔,2当量)。混合物在80摄氏度条件下反应4小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,混合物浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→20%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压浓缩除去溶剂,得到化合物46-1(白色固体,550毫克,产率64%)。MS(ESI,m/z):643.2/644.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.74–7.69(m,1H),7.56(d,J=2.7Hz,1H),7.45–7.41(m,1H),7.32–7.26(m,1H),7.11(d,J=2.7Hz,1H),5.31(s,2H),4.52–4.41(m,4H),3.71–3.65(m,2H),3.55(s,3H),2.67–2.54(m,1H),2.47–2.33(m,1H),2.06–1.96(m,2H),1.91–1.76(m,2H),1.55(s,9H),0.88–0.83(m,3H)。
步骤2
在25摄氏度,氮气保护搅拌条件下,向化合物46-1(490毫克,0.724毫摩尔,1当量),碳酸铯(496.51毫克,1.448毫摩尔,2当量)和三乙烯二胺(17.09毫克,0.145毫摩尔,0.2当量)的N,N-二甲基甲酰胺(5毫升)溶液中加入化合物35-1(118.36克,0.796毫摩尔,1.1当量)。混合物在90摄氏度条件下搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,加入200毫升水稀释反应液,水层用乙酸乙酯(300毫升 x 3)萃取,合并后有机层用饱和食盐水(100毫升 x 3)洗涤,再用无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压浓缩除去溶剂,得到化合物46-2(白色固体,360毫克,产率63%)。MS(ESI,m/z):748.4[M+H]
+。
步骤3
通过超临界液相色谱对步骤2所得化合物46-2(360毫克)进行手性拆分:手性柱CHIRAL ART Cellulose-SC,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:乙醇(0.5%,2摩尔/升氨甲醇);流速:60毫升/分钟;柱温:35摄氏度;用35%流动相B洗脱;检测器UV222纳米,得到两个产品。较短保留时间(4.85分钟)的产品为化合物46-2a(白色固体,152毫克,回收率42%),MS(ESI,m/z):748.4[M+H]
+;较长保留时间(5.16分钟)的产品为化合物46-2b(白色固体,153毫克,产率42%),MS(ESI,m/z):748.4[M+H]
+。
步骤4
在25摄氏度氮气保护条件下,依次向50毫升三口瓶中加入化合物46-2a(60毫克,0.08毫摩尔,1.0当量),1,3-二甲基巴比妥酸(18.79毫克,0.12毫摩尔,1.5当量),四(三苯基膦)钯(4.64毫克,0.004毫摩尔,0.05当量)和二氯甲烷(0.8毫升)。所得混合物在0摄氏度反应2小时,反应过程通过液质和薄色谱层层析监控。反应结束后减压浓缩除去多余的溶剂得到粗产品。粗产品通过反相快速色谱柱(C18柱)进行纯化,在20分钟内用10%→50%的乙腈/水流动相(0.1%甲酸)进行洗脱,检测器UV254纳米,得到化合物46-3a(黄色固体,50毫克,产率70%)。MS(ESI,m/z):708.3[M+H]
+。
步骤5
在25摄氏度搅拌条件下,向化合物46-3a(44毫克,0.058毫摩尔,1.00当量),化合物43-5(27.13毫克,0.07毫摩尔,1.20当量)和醋酸(5.26毫克,0.087毫摩尔,1.5当量)的甲醇(0.5毫升)溶液中加入氰基硼氢化钠(11毫克,0.174毫摩尔,3.0当量)。所得混合物在25摄氏度反应1小时,反应过程通过液质和薄色谱层层析监控。反应结束后,反应液减压浓缩得到粗产品。所得粗产品用硅胶 柱层析法纯化,用0%→5%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物46-4a(白色固体,38毫克,产率56%)。MS(ESI,m/z):1079.5[M+H]
+。
步骤6
在0摄氏度搅拌条件下,向化合物46-4a(38毫克,0.033毫摩尔,1.00当量)的甲醇(2毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,2毫升)。混合物在室温下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过高效液相色谱进行纯化,色谱柱YMC-Actus Triart C18,30 x 150毫米,5微米;流动相A:水(0.05%盐酸),流动相B:乙腈;流速:60毫升/分钟;在1.5分钟内用5%→5%的B,在7分钟内用20%→45%的B相进行洗脱,检测器UV 220纳米。得到化合物46(黄色固体,19.7毫克,产率58%)。MS(ESI,m/z):935.4[M+H]
+;
1H NMR(400MHz,DMSO-d
6)δ11.10(s,1H),10.45(s,1H),10.12(s,1H),9.73(s,1H),9.47(s,1H),7.80–7.67(m,2H),7.53–7.50(m,1H),7.41–7.32(m,2H),7.08–6.97(m,2H),6.94–6.91(m,1H),6.62(s,1H),5.12–5.02(m,1H),4.80–4.73(m,1H),4.69–4.63(m,1H),4.43–4.35(m,2H),4.16(s,2H),3.92(s,1H),3.84–3.76(m,2H),3.70–3.54(m,3H),3.50–3.46(m,3H),3.32–3.25(m,2H),3.22–3.10(m,2H),2.96–2.82(m,1H),2.60–2.55(m,1H),2.47–2.39(m,1H),2.33–2.21(m,3H),2.09–1.86(m,10H),1.81–1.70(m,2H),0.74–0.70(m,3H);
19F NMR(377MHz,DMSO-d
6)δ-116.31,-119.15,-123.10。
实施例47
(2S,4R)-1-((S)-2-(3-(3-((S)-2-((((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-7-(8-乙炔基-7-氟-3-羟基萘-1-基)-6,8-二氟喹唑啉-2-基)氧基)甲基)吡咯烷-1-基)丙氧基)丙氨基)-3,3-二甲基丁酰基)-4-羟基-N-(4-(4-甲基噻唑-5-基)苯基)吡咯烷-2-甲酰胺一甲酸盐47
步骤1
在25摄氏度氮气保护搅拌条件下,向化合物40-2(800毫克,1.14毫摩尔,1.0当量),化合物44-7(676.61毫克,1.254毫摩尔,1.1当量),3-(叔丁基)-4-(2,6-二甲氧基苯基)-2,3-二氢苯并[D][1,3]氧,膦戊轭(76.87毫克,0.228毫摩尔,0.2当量)和三(二亚苄基丙酮)二钯(106.53毫克,0.114毫摩尔,0.1当量)的甲苯/水(8毫升/1.6毫升)的混合溶液中分批加入磷酸钾(493.88毫克,2.280毫摩尔,2当量)。混合物在80摄氏度条件下反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液冷却至室温,混合物浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→8%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物47-1(黄色半固体,720毫克,产率65%)。MS(ESI,m/z):972.5[M+H]
+。
步骤2
通过超临界液相色谱对步骤3所得化合物47-1(720毫克)进行手性拆分:手性柱CHIRALPAK AD-H,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:异丙醇(0.5%,2摩尔/升氨甲醇);流速:50毫升/分钟;柱温:35摄氏度;用30%流动相B洗脱;检测器UV222纳米,得到两个产品。较短保留时间(2.00分钟)的产品为化合物47-1a(黄色固体,310毫克,回收率43%),MS(ESI,m/z):972.5[M+H]
+;较短保留时间(2.4分钟)的产品为化合物47-1b(黄色固体,300毫克,产率41%),MS(ESI,m/z):972.5[M+H]
+。
步骤3
在0摄氏度搅拌条件下,向反应瓶中依次加入化合物47-1a(280毫克,0.274毫摩尔,1当量), 高碘酸钠(298.56毫克,1.370毫摩尔,5当量),四氯化碳(1.5毫升),乙腈(1.5毫升),向其中缓慢滴加三氯化钌(6.29毫克,0.027毫摩尔,0.1当量)的水(2.25毫升)溶液。混合物在0摄氏度条件下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%碳酸氢铵);流动相B:甲醇,在30分钟内用5%→95%的B相进行洗脱;检测器UV254/220纳米;得到化合物47-2a(白色固体,90毫克,产率33%)。MS(ESI,m/z):990.5[M+H]
+。
步骤4
在室温搅拌条件下,向化合物47-2a(80毫克,0.077毫摩尔,1.00当量)的N,N-二甲基甲酰胺(1毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(35.73毫克,0.092毫摩尔,1.2当量)。混合物在25摄氏度下搅拌反应10分钟,再向反应液中加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰基]-4-羟基-N-[[4-(4-甲基-1,3-噻唑-5-基)苯基]甲基]吡咯烷-2-甲酰胺盐酸盐(43.89毫克,0.092毫摩尔,1.2当量)和N,N-二异丙基乙胺(40.08毫克,0.308毫摩尔,4当量)。混合物在25摄氏度下继续搅拌反应2小时,反应过程通过液质和薄层层析来监控。反应完全后,所得混合物通过制备薄层层析进行纯化,用0%→8%的甲醇/二氯甲烷流动相梯度洗脱,得到化合物47-3a(白色固体,80毫克,产率74%)。MS(ESI,m/z):1402.7[M+H]
+。
步骤5
在0摄氏度搅拌条件下,向化合物47-3a(78毫克,0.054毫摩尔,1.00当量)的N,N-二甲基甲酰胺(1毫升)溶液中加入氟化铯(42.23毫克,0.27毫摩尔,5.00当量)。混合物在室温下反应1小时,反应过程通过液质和薄层层析来监控。反应完全后,加入20毫升水淬灭反应,所得混合物用乙酸乙酯(20毫升 x 3)萃取。合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂;滤液减压浓缩得到粗产品。粗产品通过制备薄层层析进行纯化,用0%→8%的甲醇/二氯甲烷流动相梯度洗脱,得到化合物47-4a(白色固体,46毫克,产率62%)。MS(ESI,m/z):1246.6[M+H]
+。
步骤6
在0摄氏度搅拌条件下,向化合物47-4a(46毫克,0.035毫摩尔,1.00当量)的甲醇(1毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,1毫升)。混合物在室温下反应1小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过高效液相色谱进行纯化,色谱柱Sunfire perp C18 Column,30 x 150毫米,5微米;流动相A:水(0.1%甲酸),流动相B:乙腈;流速:60毫升/分钟;在1.5分钟内用5%→5%的B,在2分钟内用5%→16%的B,在7分钟内用16%→30%的B相进行洗脱;检测器UV 220纳米;得到化合物47(白色固体,16.8毫克,产率41%)。MS(ESI,m/z):1102.5[M+H]
+。
1H NMR(400MHz,DMSO-d
6)δ8.98(s,1H),8.58–8.55(m,1H),8.24(s,1H),7.98–7.96(m,1H),7.87(d,J=9.6Hz,1H),7.55–7.45(m,2H),7.42–7.36(m,5H),7.15(d,J=2.4Hz,1H),4.53(d,J=9.6Hz,1H),4.45–4.39(m,2H),4.35–4.30(m,2H),4.27–4.19(m,3H),4.04–3.99(m,1H),3.96(s,1H),3.67–3.63(m,1H),3.62–3.58(m,4H),3.56–3.52(m,2H),3.50–3.46(m,3H),3.39–3.36(m,2H),3.04–3.00(m,1H),2.91–2.84(m,1H),2.82–2.75(m,1H),2.56–2.52(m,1H),2.43(s,3H),2.37–2.23(m,2H),2.19–2.13(m,1H),2.05–2.00(m,1H),1.92–1.85(m,2H),1.79–1.58(m,9H),0.89(s,9H);
19F NMR(377MHz,DMSO-d
6)δ-110.19,-118.96,-124.73。
实施例48
(2S,4R)-1-((S)-2-(3-(2-((2R,6R,7aS)-7a-((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-7-(8-乙基-7-氟-3-羟基萘-1-基)-6,8-二氟喹唑啉-2-基)氧基)甲基)-6-氟六氢-1H-吡咯嗪-2-基)乙氧基)丙胺基)-3,3-二甲基丁酰)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺48
步骤1
在0摄氏度氮气保护搅拌条件下,将化合物3-丁烯-1-醇(17.13克,225.62毫摩尔,1.1当量)溶于300毫升四氢呋喃中,向其中分批加入氢化钠(60%,24.61克,615.33毫摩尔,3.0当量),混合物在0摄氏度反应30分钟后,将溴乙酸(30克,205.11毫摩尔,1.0当量)缓慢滴加上述混合体系中。混合物在70摄氏度条件下反应2小时,反应过程通过薄层层析来监控。反应结束后,反应液冷却至室温,倒入水溶液中淬灭,用2摩尔盐酸调节pH至1~3,乙酸乙酯萃取,合并有机相后干燥,过滤除去干燥剂,滤液浓缩得到粗产品化合物48-1(黄色油,30克,产率56%),粗产品直接用于下一步合成。
步骤2
在0摄氏度氮气保护搅拌条件下,将化合物48-1(30克,218.99毫摩尔,1.0当量)溶于300毫升无水四氢呋喃中,向其中缓慢滴加四氢铝锂的四氢呋喃溶液(2.5摩尔每升,87.60毫升,218.99毫摩尔,1.0当量),所得混合物在25摄氏度反应2小时,反应过程通过薄层层析来监控。反应结束后,反应液冷却至0摄氏度,向反应液中依次缓慢滴加水(20毫升)、20%的氢氧化钠(20毫升)和水(60毫升),加完继续搅拌30分钟,混合物过滤,滤液浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→50%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物48-2(无色油,5克,产率30%)。
1H NMR(400MHz,CDCl
3)δ5.88-5.77(m,1H),5.20–4.99(m,2H),3.74-3.71(m,2H),3.63–3.48(m,4H),2.38-2.32(m,2H)。
步骤3
在0摄氏度氮气保护搅拌条件下,将化合物48-2(5克,43.04毫摩尔,1.0当量)和四溴化碳(15.66克,47.34毫摩尔,1.1当量)溶于50毫升二氯甲烷中,缓慢加入三苯基膦(12.40克,47.34毫摩尔,1.1当量),混合物在0摄氏度反应1小时,反应过程通过薄层层析来监控。反应结束后,反应液浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物48-3(无色油,3克,产率50%)。
1H NMR(400MHz,CDCl
3)δ5.90–5.75(m,1H),5.14–5.00(m,2H),3.80–3.69(m,2H),3.60–3.51(m,2H),3.50–3.42(m,2H),2.41–2.31(m,2H)。
步骤4
在0摄氏度氮气保护搅拌条件下,向反应瓶中依次加入化合物48-4(参考WO2021041671制得,2克,8.82毫摩尔,1.0当量),无水甲醇(0.57克,17.65毫摩尔,2当量)以及20毫升无水四氢呋喃,随后向上述溶液中分批加入硼氢化锂(0.40克,17.65毫摩尔,2当量),所得混合物在25摄氏度反应2小时,反应过程通过薄层层析来监控。反应结束后,反应液倒入饱和碳酸氢钠溶液中淬灭,用三氯甲烷和异丙醇混合溶剂(3/1)萃取,有机相浓缩得到粗产品化合物48-5(无色油,1.5克,产率90%)。MS(ESI,m/z):174.1[M+H]
+。粗产品直接用于下一步合成。
步骤5
在0摄氏度氮气保护搅拌条件下,向反应瓶中依次加入化合物48-5(1.5克,8.22毫摩尔,1.0当量),咪唑(0.88克,12.34毫摩尔,1.5当量)以及20毫升二氯甲烷,随后向其中缓慢加入叔丁基二苯基氯硅烷(3.09克,10.69毫摩尔,1.3当量),所得混合物在25摄氏度反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液倒入饱和碳酸氢钠溶液中淬灭,用二氯甲烷(20毫升 x 3)萃取,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→5%的甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物48-5(白色固体,3克,产率84%)。MS(ESI,m/z):412.2[M+H]
+;
1H NMR(400MHz,CDCl
3)δ7.64–7.61(m,4H),7.46–7.37(m,6H),5.33-5.18(m,1H),4.18–4.09(m,1H),3.57(d,J=10.0Hz,1H),3.43(d,J=10.0Hz,1H),3.15–3.02(m,1H),2.76–2.67(m,1H),2.42–2.12(m,3H),2.04–1.92(m,2H),1.05(s,9H)。
步骤6
在零下78摄氏度氮气保护搅拌条件下,将化合物48-6(2.6克,6.32毫摩尔,1.0当量)溶于30毫升四氢呋喃中,向其中缓慢滴加二异丙基氨基锂的四氢呋喃溶液(1摩尔每升,8.2毫升,8.20 毫摩尔,1.3当量),混合物在零下78摄氏度反应30分钟后,向上述体系中缓慢滴加48-3(1.46克,8.20毫摩尔,1.3当量)的无水四氢呋喃(2毫升)溶液,加完缓慢升至室温,反应过程通过液质和薄层层析来监控。反应结束后,反应液倒入饱和氯化铵溶液中淬灭,用乙酸乙酯萃取三次,合并有机相,有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→30%的乙酸乙酯/石油醚流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物48-7(白色固体,1.9克,产率55%)。MS(ESI,m/z):510.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.65–7.61(m,4H),7.45–7.39(m,6H),5.87–5.69(m,1H),5.36–5.18(m,1H),5.10–4.93(m,2H),4.25–4.07(m,1H),3.52–3.39(m,5H),3.19–3.01(m,1H),2.97–2.82(m,1H),2.41–2.14(m,5H),2.03–1.87(m,1H),1.77–1.68(m,1H),1.59–1.44(m,2H),1.04(s,9H)。
步骤7
通过超临界液相色谱对本实施例步骤6所得化合物48-7(1.9克)进行异构体分离:手性柱CHIRALPAK IA,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:甲醇;流速:50毫升/分钟;柱温:35摄氏度;用15%流动相B洗脱;检测器UV225纳米,得到两个产品。较短保留时间(6.83分钟)的产品为化合物48-7a(无色油,1.4克,回收率80%),化合物48-7a:MS(ESI,m/z):510.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.69–7.57(m,4H),7.51–7.32(m,6H),5.86–5.69(m,1H),5.38–5.13(m,1H),5.09–4.89(m,2H),4.24–4.05(m,1H),3.59(d,J=10.3Hz,1H),3.54–3.38(m,5H),3.22–3.00(m,1H),2.97–2.81(m,1H),2.45–2.11(m,5H),2.05–1.91(m,1H),1.76–1.66(m,1H),1.57–1.43(m,1H),1.04(s,9H);
19F NMR(282MHz,CDCl
3)δ-174.00。较长保留时间(10.02分钟)的产品为化合物48-7b(无色油,270毫克,回收率16%),化合物48-7b:MS(ESI,m/z):510.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.67–7.56(m,4H),7.48–7.35(m,6H),5.86–5.67(m,1H),5.47–5.21(m,1H),5.10–4.93(m,2H),4.21–4.01(m,1H),3.59–3.53(m,1H),3.52–3.36(m,5H),3.36–3.20(m,1H),2.75–2.63(m,1H),2.58–2.40(m,1H),2.32–2.12(m,3H),2.08–1.89(m,2H),1.81–1.74(m,1H),1.65–1.51(m,1H),1.04(s,9H);
19F NMR(282MHz,CDCl
3)δ-170.22。
步骤8
在0摄氏度氮气保护搅拌条件下,将化合物48-7a(600毫克,1.11毫摩尔,1.0当量)溶于6毫 升四氢呋喃中,向其中缓慢滴加氢化铝锂的四氢呋喃溶液(1摩尔每升,1.67毫升,1.67毫摩尔,1.5当量),混合物在60摄氏度反应2小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液降至0摄氏度,向其中依次缓慢加入水(1毫升)、20%的氢氧化钠水溶液(1毫升)和水(3毫升)淬灭,所得混合物过滤,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%的氨甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物48-8a(无色油,210毫克,产率69%)。MS(ESI,m/z):258.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ5.83–5.66(m,1H),5.27–4.91(m,3H),3.42–3.17(m,7H),3.13–3.03(m,1H),2.99–2.79(m,1H),2.60–2.53(m,1H),2.31–2.08(m,3H),2.06–1.79(m,3H),1.66–1.50(m,3H)。
步骤9
在25摄氏度氮气保护搅拌条件下,向反应瓶中依次加入化合物46-1(600毫克,0.739毫摩尔,1当量),化合物48-8a(200毫克,0.739毫摩尔,1当量),碳酸铯(506.64毫克,1.478毫摩尔,2当量),三乙烯二胺(17.44毫克,0.148毫摩尔,0.2当量)以及无水N,N-二甲基甲酰胺(5毫升)。所得混合物在80摄氏度条件下反应2.5小时,反应过程通过液质和薄层层析来监控。反应完全后,加入50毫升水稀释反应液,水层用乙酸乙酯(50毫升 x 3)萃取,合并后有机层用饱和食盐水(50毫升 x 3)洗涤,无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压浓缩除去溶剂,得到化合物48-9a(白色固体,580毫克,产率63%)。MS(ESI,m/z):864.4[M+H]
+;
1H NMR(300MHz,CDCl
3)δ7.71–7.65(m,1H),7.52(d,J=3.0Hz,1H),7.40–7.26(m,1H),7.26–7.22(m,1H),7.11(d,J=3.0Hz,1H),5.84–5.56(m,3H),5.29(s,2H),5.10–4.94(m,2H),4.50-4.0(m,6H),3.70–3.50(m,5H),3.48–3.34(m,5H),2.77–2.53(m,2H),2.46–2.24(m,6H),2.14–1.83(m,7H),1.52(s,9H),1.32–1.22(m,2H),0.86–0.80(m,3H)。
步骤10
通过超临界液相色谱对本实施例步骤9所得化合物48-9a(580毫克)进行手性拆分:手性柱CHIRAL ART Amylose-SA,3 x 25厘米,5微米;流动相A:超临界二氧化碳,流动相B:异丙醇(0.5%,2摩尔/升氨甲醇);流速:100毫升/分钟;柱温:35摄氏度;用40%流动相B洗脱;检测器UV222纳米,得到两个产品。较短保留时间(2.75分钟)的产品为化合物48-9aa(白色固体,220毫克,回收率38%);MS(ESI,m/z):864.4[M+H]
+;较长保留时间(4.37分钟)的产品为化合物48-9ab(白色固体,230毫克,产率39%);MS(ESI,m/z):864.4[M+H]
+。
步骤11
在0摄氏度搅拌条件下,向反应瓶中依次加入化合物48-9aa(120毫克,0.132毫摩尔,1当量),高碘酸钠(178.24毫克,0.792毫摩尔,6当量),四氯化碳(0.6毫升),乙腈(0.6毫升),随后向其中缓慢滴加三氯化钌(3.13毫克,0.013毫摩尔,0.1当量)的水(0.9毫升)溶液。所得混合物在0摄氏度条件下反应1小时,反应过程通过液质来监控。反应结束后,反应液直接浓缩得到粗产品。所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%碳酸氢铵);流动相B:甲醇,在25分钟内用5%→95%的B相进行洗脱;检测器UV254/220纳米;得到化合物48-10aa(白色固体,70毫克,产率54%)。MS(ESI,m/z):882.4[M+H]
+。
步骤12
在室温搅拌条件下,向化合物48-11aa(70毫克,0.071毫摩尔,1.00当量)的N,N-二甲基甲酰胺(1.5毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(37.17毫克,0.092毫摩尔,1.3当量)。混合物在25摄氏度下搅拌反应10分钟后,向反应液中依次加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(40.11毫克,0.085毫摩尔,1.2当量)和N,N-二异丙基乙胺(38.87毫克,0.284毫摩尔,4当量)。混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质来监控。反应完全后,所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%碳酸氢铵);流动相B:甲醇,在25分钟内用10%→95%的B相进行洗脱;检测器UV254/220纳米;得到化合物48-11aa(白色固体,80毫克,产率81%)。MS(ESI,m/z):1308.6[M+H]
+。
步骤13
在0摄氏度搅拌条件下,向化合物48-11aa(80毫克,0.058毫摩尔,1.00当量)的甲醇(2毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,2毫升)。混合物在室温下反应2小时,反应过程通过液质来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%碳酸氢铵);流动相B:甲醇,在30分钟内用70%→95%的B相进行洗脱;检测器UV254/220纳米。得到化合物48(白色固体,50毫克,产率70%)。MS(ESI,m/z):1164.5[M+H]
+。
1H NMR(400MHz,DMSO-d
6)δ10.02(s,1H),8.98(s,1H),8.36(d,J=7.8Hz,1H),7.89–7.74(m,2H),7.70–7.62(m,1H),7.45–7.30(m,6H),7.01(d,J=2.6Hz,1H),5.36–5.22(m,1H),5.11(s,1H),4.97–4.84(m,1H),4.51(d,J=9.3Hz,1H),4.47–4.37(m,1H),4.33–4.19(m,3H),4.12–4.01(m,1H),4.00–3.91(m,1H),3.64–3.43(m,8H),3.42–3.34(m,2H),3.30–3.25(m,1H),3.24–3.17(m,1H),3.12– 2.87(m,2H),2.62–2.54(m,1H),2.45(s,3H),2.39–1.94(m,8H),1.82–1.73(m,1H),1.65(s,4H),1.58–1.50(m,2H),1.50–1.41(m,1H),1.37(d,J=7.0Hz,3H),0.90(s,9H),0.79–0.70(m,3H);
19F NMR(377MHz,DMSO-d
6)δ-118.64,-119.23,-123.86,-171.35。
实施例49
(2S,4R)-1-((S)-2-(3-(2-((2S,6R,7aS)-7a-((S或R)-4-((1R,5S)-3,8-二氮杂环[3.2.1]辛烷-3-基)-7-(8-乙基-7-氟-3-羟基萘-1-基)-6,8-二氟喹唑啉-2-基)氧基)甲基)-6-氟六氢-1H-吡咯嗪-2-基)乙氧基)丙胺基)-3,3-二甲基丁酰)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺49
步骤1
在0摄氏度氮气保护搅拌条件下,将化合物48-7b(270毫克,0.503毫摩尔,1.0当量)溶于3毫升四氢呋喃中,随后向其中缓慢滴加氢化铝锂的四氢呋喃溶液(1摩尔每升,0.75毫升,0.75毫摩尔,1.5当量),所得混合物在60摄氏度反应16小时,反应过程通过液质和薄层层析来监控。反应结束后,反应液降至0摄氏度,向其中依次缓慢加入水(1毫升)、20%的氢氧化钠水溶液(1毫升)和水(3毫升)淬灭,混合物过滤,滤液浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%的氨甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压旋蒸除去溶剂,得到化合物49-1(无色油,60毫克,产率44%)。MS(ESI,m/z):258.2[M+H]
+;
1H NMR(300MHz,CDCl
3)δ5.97–5.69(m,1H),5.39–4.95(m,3H),3.53–3.30(m,5H),3.29–3.19(m,1H),3.15–2.99(m,2H),2.81–2.69(m,1H),2.63–2.45(m,1H),2.41–2.20(m,3H),2.18–1.89(m,3H),1.70–1.56(m,2H),1.55–1.43(m,1H)。
步骤2
在25摄氏度氮气保护搅拌条件下,向反应瓶中依次加入化合物46-1(120毫克,0.177毫摩尔,1当量),化合物49-1(48.02毫克,0.177毫摩尔,1当量),碳酸铯(121.59毫克,0.354毫摩尔,2当量),三乙烯二胺(4.19毫克,0.035毫摩尔,0.2当量)以及无水N,N-二甲基甲酰胺(2毫升)。所得混合物在80摄氏度条件下搅拌反应3小时,反应过程通过液质和薄层层析来监控。反应完全后,加入20毫升水稀释反应液,所得混合物用乙酸乙酯(20毫升 x 3)萃取,合并后有机层用饱和食盐水(20毫升 x 3)洗涤,无水硫酸钠干燥。过滤除去干燥剂,滤液减压浓缩得到粗产品。粗产品通过硅胶柱层析法纯化,用0%→10%甲醇/二氯甲烷流动相梯度洗脱,所得馏分通过减压浓缩除去溶剂,得到化合物49-2(白色固体,120毫克,产率73%)。MS(ESI,m/z):864.4[M+H]
+。
步骤3
通过超临界液相色谱对本实施例步骤2所得化合物49-2(120毫克)进行手性拆分:手性柱CHIRAL ART Amylose-SA,3 x 25厘米,5微米;流动相A:正己烷(10毫摩尔/升氨甲醇),流动相B:乙醇;流速:20毫升/分钟;在11分钟内用30%流动相B洗脱;检测器UV230/220纳米,得到两个产品。较短保留时间(5.39分钟)的产品为化合物49-2a(白色固体,58毫克,回收率48%),MS(ESI,m/z):864.4[M+H]
+;较长保留时间(7.33分钟)的产品为化合物49-2b(白色固体,50毫克,回收率41%),MS(ESI,m/z):864.4[M+H]
+。
步骤4
在0摄氏度搅拌条件下,向反应瓶中依次加入化合物49-2a(60毫克,0.064毫摩尔,1当量),高碘酸钠(86.15毫克,0.384毫摩尔,6当量),四氯化碳(0.5毫升),乙腈(0.5毫升),向其中缓慢滴加三氯化钌(1.51毫克,0.006毫摩尔,0.1当量)的水(0.75毫升)溶液。混合物在0摄氏度条件下反应1小时,反应过程通过液质来监控。反应结束后,反应液倒入水中淬灭,氯仿和异丙醇混合溶剂(3/1)萃取三次,合并后有机相用无水硫酸钠干燥,过滤除去干燥剂,滤液减压浓缩得到粗产品。所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%碳酸氢铵);流动相B:甲醇,在25分钟内用5%→95%的B相进行洗脱;检测器UV254/220纳米;得到化合物49-3a(白色固体,20毫克,产率34%)。MS(ESI,m/z):882.4[M+H]
+。
步骤5
在室温搅拌条件下,向化合物49-3a(70毫克,0.022毫摩尔,1.00当量)的N,N-二甲基甲酰胺(1毫升)溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯(10.35毫克,0.026毫摩尔,1.3当量)。混合物在25摄氏度下搅拌反应10分钟后,向反应液中依次加入(2S,4R)-1-[(2S)-2-氨基-3,3-二甲基丁酰]-4-羟基-N-[(1S)-1-[4-(4-甲基-1,3-噻唑-5-基)苯基]乙基]吡咯烷-2-甲酰胺盐酸盐(11.09毫克,0.024毫摩尔,1.1当量)和N,N-二异丙基乙胺(11.72毫克,0.088毫摩尔,4当量)。所得混合物在25摄氏度下继续搅拌反应1小时,反应过程通过液质来监控。反应完全后,所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%碳酸氢铵);流动相B:甲醇,在25分钟内用10%→95%的B相进行洗脱;检测器UV254/220纳米;所得馏分减压浓缩得到化合物49-4a(白色固体,20毫克,产率84%)。MS(ESI,m/z):1308.6[M+H]
+。
步骤6
在0摄氏度搅拌条件下,向化合物49-4a(20毫克,0.015毫摩尔,1.00当量)的甲醇(1毫升)溶液中滴加盐酸的1,4-二氧六环溶液(4摩尔/升,1毫升)。混合物在室温下反应2小时,反应过程通过液质来监控。反应结束后,反应液浓缩得到粗产品。所得粗产品通过反相色谱(C18柱)进行纯化,流动相A:水(0.1%碳酸氢铵);流动相B:甲醇,在30分钟内用70%→95%的B相进行洗脱;检测器UV254/220纳米;所得馏分减压浓缩得到化合物49(白色固体,5.5毫克,产率30%)。MS(ESI,m/z):1164.5[M+H]
+。
1H NMR(400MHz,CD
3OD)δ8.86(s,1H),7.69–7.55(m,2H),7.47–7.33(m,4H),7.31–7.17(m,2H),6.96(d,J=2.7Hz,1H),5.33–5.20(m,1H),5.04–4.96(m,1H),4.64(s,1H),4.60–4.53(m,1H),4.50–4.38(m,3H),4.30–4.20(m,2H),3.85(d,J=11.0Hz,1H),3.77–3.70(m,1H),3.70–3.64(m,2H),3.64–3.54(m,4H),3.53–3.43(m,3H),3.39–3.34(m,1H),3.17–3.04(m,2H),2.92–2.78(m,1H),2.68–2.49(m,4H),2.48–2.41(m,4H),2.26–2.17(m,1H),2.17–2.03(m,2H),1.97–1.89(m,1H),1.88–1.81(m,3H),1.75–1.63(m,3H),1.59–1.44(m,3H),1.36–1.30(m,1H),1.03(s,9H),0.84–0.69(m,3H);
19F NMR(377MHz,CD
3OD)δ-118.75,-121.17,-124.29,-174.68。
效果实施例1
1.实验目的
通过基于KRAS_G12D与SOS1结合的药物筛选体系来检测小分子化合物对于KRAS-G12D与SOS1结合活性的抑制能力。
2.实验材料及仪器设备
试剂 | 品牌 | 货号 |
KRAS-G12D/SOS1 binding kits | Cisbio | 63ADK000CB21PEH |
GTP | Sigma | V900868 |
耗材 | 品牌 | 货号 |
Topseal A | PerkinElmer | E5341 |
384-Well Polypropylene microplate | labcyte | PP-0200 |
96 Well Plates | Nunc | 249944 |
384-well plates | Corning | CLS4514 |
仪器 | 品牌 | 货号 |
Envision | Perkin Elmer | 2104 |
Centrifuge | Eppendorf | 5810R |
Multi-channel pipettes | Eppendorf/Sartorius | / |
Echo | Labcyte | / |
3.实验方法
3.1实验步骤:
a)BI-2852作为阳性对照,其储存液为稀释的第一个点,3倍稀释,稀释10+0个点。同样待测化合物稀释的第一个点也为其储存液,3倍稀释,稀释11+0个点。用Echo转移0.2μL梯度稀释的化合物溶液到384孔板中,每个化合物做2个复孔,DMSO终浓度均为1%。1000rpm/min,离心1min。Reference终浓度为100,33.33,11.11,3.70,1.23,0.412,0.137,0.046,0.015,0.005,0μM。待测化合物终浓度200,66.67,22.22,7.41,2.47,0.27,0.091,0.03,0.0152,0.01,0μM。
b)将试剂盒中的KRAS_G12D与终浓度为10μM GTP在稀释液中共同配制,转移5μL到384反应板中,1000rpm/min,离心1min,
c)转移5μL SOS1混合液到384反应板中,1000rpm/min,离心1min,25℃孵育15min。
d)转移10μL检测混合液到384反应板中,1000rpm/min,离心1min,4℃孵育过夜。
e)使用Envision多功能读板机读取激发波长665nm和发射波长615nm。665/615 Ratio信号强度用于表征酶的活性程度。
f)分析原始数据。
3.2实验数据处理方法:
通过Graphpad Prism 8非线性回归方程拟合化合物IC50:
阴性对照:DMSO
阳性对照:100μM BI-2852
利用以下非线性拟合公式来得到化合物的IC
50(半数抑制浓度):
Y=Bottom+(Top-Bottom)/(1+10^((LogIC
50-X)*HillSlope))
X:化合物浓度log值
Y:665/615 Ratio
效果实施例B
1.实验目的
通过基于KRAS_G12D与cRAF结合的药物筛选体系来检测小分子化合物对于KRAS_G12D与 cRAF结合活性的抑制能力。
2.实验材料及仪器设备
试剂 | 品牌 | 货号 |
KRAS-G12D/cRAF binding kits | Cisbio | 63ADK000CB21PEG |
GTP | Sigma | V900868 |
耗材 | 品牌 | 货号 |
Topseal A | PerkinElmer | E5341 |
384-Well Polypropylene microplate | labcyte | PP-0200 |
96 Well Plates | Nunc | 249944 |
384-well plates | Corning | CLS4514 |
仪器 | 品牌 | 货号 |
Envision | Perkin Elmer | 2104 |
Centrifuge | Eppendorf | 5810R |
Multi-channel pipettes | Eppendorf/Sartorius | / |
Echo | Labcyte | / |
3.实验方法
3.1实验步骤:
a)BI-2852作为阳性对照,其储存液为稀释的第一个点,3倍稀释,稀释10+0个点。同样待测化合物稀释的第一个点也为其储存液,3倍稀释,稀释11+0个点。用Echo转移0.2μL梯度稀释的化合物溶液到384孔板中,每个化合物做2个复孔,DMSO终浓度均为1%。1000rpm/min,离心1min。阳性对照的终浓度为100,33.33,11.11,3.70,1.23,0.412,0.137,0.046,0.015,0.005,0μM。待测化合物终浓度200,66.67,22.22,7.41,2.47,0.27,0.091,0.03,0.0152,0.01,0μM。
b)将试剂盒中的KRAS_G12D与终浓度为10μM GTP在稀释液中共同配制,转移5μL到384反应板中,1000rpm/min,离心1min,
c)转移5μL cRAF混合液到384反应板中,1000rpm/min,离心1min,25℃孵育15min。
d)转移10μL检测混合液到384反应板中,1000rpm/min,离心1min,4℃孵育过夜。
e)使用Envision多功能读板机读取激发波长665nm和发射波长615nm。665/615 Ratio信号强度用于表征酶的活性程度。
f)分析原始数据。
3.2实验数据处理方法:
通过Graphpad Prism 8非线性回归方程拟合化合物IC
50:
阴性对照:DMSO
阳性对照:100μM BI-2852
利用以下非线性拟合公式来得到化合物的IC50(半数抑制浓度):
Y=Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope))
X:化合物浓度log值
Y:665/615 Ratio
效果实施例C
1实验目的
通过基于KRAS_WT与SOS1结合的药物筛选体系来检测小分子化合物对于KRAS_WT与SOS1结合活性的抑制能力。
2实验材料及仪器设备
试剂 | 品牌 | 货号 |
KRAS-WT/SOS1 binding kits | Cisbio | 63ADK000CB15PEH |
GTP | Sigma | V900868 |
耗材 | 品牌 | 货号 |
Topseal A | PerkinElmer | E5341 |
384-Well Polypropylene microplate | labcyte | PP-0200 |
96Well Plates | Nunc | 249944 |
384-well plates | Corning | CLS4514 |
仪器 | 品牌 | 货号 |
Envision | Perkin Elmer | 2104 |
Centrifuge | Eppendorf | 5810R |
Multi-channel pipettes | Eppendorf/Sartorius | / |
Echo | Labcyte | / |
3实验方法
3.1实验步骤:
a)BI-2852作为阳性对照,其储存液为稀释的第一个点,3倍稀释,稀释10+0个点。同样待测化合物稀释的第一个点也为其储存液,3倍稀释,稀释11+0个点。用Echo转移0.2μL梯度稀释的化合物溶液到384孔板中,每个化合物做2个复孔,DMSO终浓度均为1%。1000rpm/min,离心1 min。阳性对照终浓度为100,33.33,11.11,3.70,1.23,0.412,0.137,0.046,0.015,0.005,0μM。待测化合物终浓度200,66.67,22.22,7.41,2.47,0.27,0.091,0.03,0.0152,0.01,0μM。
b)将试剂盒中的KRAS_WT与终浓度为10μM GTP在稀释液中共同配制,转移5μL到384反应板中,1000rpm/min,离心1min,
c)转移5μL SOS1混合液到384反应板中,1000rpm/min,离心1min,25℃孵育15min。
d)转移10μL检测混合液到384反应板中,1000rpm/min,离心1min,4℃孵育过夜。
e)使用Envision多功能读板机读取激发波长665nm和发射波长615nm。665/615 Ratio信号强度用于表征酶的活性程度。
f)分析原始数据。
3.2实验数据处理方法:
通过Graphpad Prism 8非线性回归方程拟合化合物IC
50:
阴性对照:DMSO
阳性对照:100μM BI-2852
利用以下非线性拟合公式来得到化合物的IC
50(半数抑制浓度):
Y=Bottom+(Top-Bottom)/(1+10^((LogIC
50-X)*HillSlope))
X:化合物浓度log值
Y:665/615 Ratio
实验结果:以上效果实施例A-C试验结果如表1所示:
效果实施例2 KRAS-G12D PROTAC分子降解试验(Degradation Assay)
2.1实验目的
通过Jess WB检测PANC-1和GP2D细胞(KRAS-G12D突变株)中的KRAS蛋白的降解情况来检测KRAS-G12D PROTAC分子对KRAS-G12D蛋白的靶向降解能力。
2.2实验材料及仪器设备
2.2实验方法
1、复苏的PANC-1、GP2D细胞贴壁培养1-2代之后,分别接种细胞到12孔板中,将细胞板置于37℃5%CO
2孵箱中培养过夜。加入实施例25的化合物,化合物浓度为30uM,1/3稀释,7+0dose,48h。
2、吸弃培养液,加入1mL 1x PBS洗涤细胞一次,每孔加入300μL Tryple(Gibco),37℃消化5min,每孔加入900μL培养液终止消化,将细胞转移至1.5mL离心管,于4℃下2000rpm离心5min收集细胞。用1mL 1x PBS洗涤细胞一次,于4℃下2000rpm离心5min收集细胞。
3、根据细胞量加入适量含有1x PMSF的裂解液,混匀后置于冰上裂解30min,于4℃13000rpm离心20min,将离心后的上清转移至新的离心管中。
4、取20μL已知不同浓度的牛血清白蛋白(BSA)以制备蛋白定量标准曲线;待测样品蛋白,按10倍稀释后,取20μL加入相应的检测孔;取Pierce
TM BCA Protein Assay Kit BCA定量试剂,200μL/well,避免产生气泡,37℃避光孵育30min后,检测波长562nm条件下的光密度(OD)值。
5、向蛋白样品中加入适量0.1x上样缓冲液(sample buffer),沸水浴加热5min,充分变性蛋白,通过Jess WB系统定量分析KRAS-G12D蛋白的降解率,其中一抗选用KRAS mouse Antibody(Lsbio),GAPDH mouse mAb(CST)。
2.3实验结果:
本发明的化合物是基于相同的WB降解实验来测定它们在细胞中的降解活性,结果通过DC
50和Dmax来展示,其中Dmax是观察到的最大蛋白降解水平,DC
50是达到Dmax的50%所需的化合物浓度;结果见表2:
表2化合物降解活性数据
实施例25分子在30μM浓度条件下,对PANC-1细胞显示大约50%的KRAS G12D蛋白降解,对GP2D细胞上显示大约40%的KRAS G12D蛋白降解。
实施例45分子在PANC-1细胞中对KRAS蛋白的DC50为53.02nM,Dmax为89%;在GP2D细胞中对KRAS蛋白的DC50为807nM,Dmax为83%。
Claims (15)
- 一种如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:其中,R 1a、R 1b和R 1c独立地为C 6-C 18芳基、被R 1a-1取代的C 6-C 18芳基或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 1a-1独立地为羟基、卤素、C 1-C 12烷基或C 2-C 6炔基;R 2a、R 4a、R 2b、R 4b、R 2c和R 5c独立地为H或卤素;R 3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 4c为H、3-6元的杂环烷基或被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基,所述的3-6元的杂环烷基中的杂原子和被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个;R 4c-1和R 4c-2独立地为C 1-C 4烷基;A a和A b独立地为CR 5a或N;R 5a独立地为H、C 1-C 4烷基或卤素;n1为1、2或3;M 1为3-9元杂环烷基、被M 1-1取代的3-9元杂环烷基、-3-9元杂环烷基-(CH 2) n1’OC(=O)-或不存在;所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH 2) n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个;n1’为1、2或3;M 1-1为C 1-C 4烷基或卤素;Y 1为O、-NR 8或不存在;环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;n2为0、1、2或3;Y 2为O、-NR 9或不存在;R 8和R 9独立地为C 1-C 4烷基;n3为1、2或3;n3’为0、1、2或3;R 10和R 11独立地为C 1-C 4烷基;环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;L a、L b和L c为连接基团;Q a、Q b和Q c为E3泛素连接酶配体。
- 如权利要求1所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物,其特征在于,所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物满足如下条件中的一个或多个:(1)R 1a、R 1b和R 1c中,所述的C 6-C 18芳基或所述的被R 1a-1取代的C 6-C 18芳基中的C 6-C 18芳基为苯基、萘基、菲基或蒽基,优选为萘基;(2)R 1a、R 2a、R 4a、R 2b、R 4b、R 2c、R 5a和R 5c中,所述的卤素为F、Cl、Br或I,例如F或Cl;(3)R 1a中,所述的C 1-C 12烷基为C 1-C 6烷基,还可以为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基;(4)R 1a中,所述的C 2-C 6炔基为C 2-C 4炔基,还可以为乙炔基;(5)R 3a中,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子为N,个数为2个,还可以为二氮杂双环[2.2.1]庚烷基、二氮杂双环[3.2.1]辛烷基或二氮杂双环[2.2.2]辛烷基;(6)R 3a中,所述的7-12元的桥环杂环烷基通过杂原子与嘧啶环相连接;(7)R 4c中,所述的3-6元的杂环烷基或所述的被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基中的3-6元的杂环烷基为氮杂环丁烷基、吡咯烷基或哌啶基;(8)R 4c中,所述的3-6元的杂环烷基或所述的被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基中的3-6元的杂环烷基通过杂原子与嘧啶环相连接;(9)R 4c-1和R 4c-2中,所述的C 1-C 4烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基,例如甲基;(10)M 1中,所述的3-9元杂环烷基为3-6元单环杂环烷基,杂原子独立地为N和/或O,杂原子个数为1或2个,进一步为吡咯烷基;(11)M 1中,所述的被M 1-1取代的3-9元杂环烷基中的3-9元杂环烷基为3-6元单环杂环烷基或8-9元并环杂化烷基,杂原子独立地为N和/或O,杂原子个数为1个或2个,进一步为吡咯烷基或六氢-1H-吡咯嗪基;(13)M 1-1中,所述的C 1-C 4烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基,例如甲基;(14)M 1-1中,所述的卤素为F、Cl、Br或I,例如F;(15)环D中,所述的3-6元的杂环烷基为氮杂环丁烷基、吡咯烷基或哌嗪;(16)R 5a、R 8、R 9、R 10、R 11和M 1-1中,所述的C 1-C 4烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基,例如甲基或乙基;(17)环E中,所述的3-6元的杂环烷基为氮杂环丁烷基;n4、n6和n7独立地为0、1或2;n5和n8独立地为1-5中任一整数,例如1、2、3、4或5;Y 3为NH、CH 2或不存在;n9为0-13中任一整数;n10和n13独立地为0、1或2;n11和n12独立地为1-7中任一整数,例如1、2、3、4或5;Z为CH 2或N;Y 4为-C(=O)或不存在;Y 5为3-6元的环烷基、3-6元的杂环烷基或5-6元的杂芳基,所述的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1、2或3个,所述的5-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1、2或3个;n14和n15独立地为0、1、2、3、4或5;n16和n19为0、1或2;n17、n18和n20独立地为0、1、2、3、4或5;Z 1为CH或N;
- 如权利要求2所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物,其特征在于,所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物满足如下条件中的一个或多个:(4)R 4c中,所述的被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基为被1个-N(CH 3) 2)取代的氮杂环丁烷基;(6)Y 5中,所述的3-6元的环烷基为环丙基、环丁基、环戊基或环己基,例如环己基;(7)Y 5中,所述的3-6元的杂环烷基为杂原子为O或S,个数为1个的5元的杂环烷基,例如呋喃基;(8)Y 5中,所述的5-6元的杂芳基为杂原子为N,个数为3个的5元杂芳基,例如1-H,1,2,3-三唑基。
- 如权利要求1所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物,其特征在于,所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、 其代谢产物或其同位素化合物满足如下任一条件:
- 如权利要求1所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物,其特征在于,其为任一方案:方案1:所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:其中,R 1a、R 1b和R 1c独立地为C 6-C 18芳基、被R 1a-1取代的C 6-C 18芳基或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 1a-1独立地为羟基、卤素、C 1-C 12烷基或C 2-C 6炔基;R 2a、R 4a、R 2b、R 4b、R 2c和R 5c独立地为H或卤素;R 3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 4c为H、3-6元的杂环烷基或被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基,所述的3-6元的杂环烷基中的杂原子和被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个;R 4c-1和R 4c-2独立地为C 1-C 4烷基;A a和A b独立地为CR 5a或N;R 5a为H、C 1-C 4烷基或卤素;n1为1、2或3;M 1为3-9元杂环烷基、被M 1-1取代的3-9元杂环烷基、-3-9元杂环烷基-(CH 2) n1’OC(=O)-或不存在;所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH 2) n1’中的杂原子独立地为N、O或S中的一 种或多种,杂原子个数为1-3个;n1’为1、2或3;M 1-1为C 1-C 4烷基;Y 1为O、-NR 8或不存在;环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;n2为0、1、2或3;Y 2为O、-NR 9或不存在;R 8和R 9独立地为C 1-C 4烷基;n3为1、2或3;n3’为0、1、2或3;R 10和R 11独立地为C 1-C 4烷基;环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;L a、L b和L c为连接基团;Q a、Q b和Q c为E3泛素连接酶配体;方案2:所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:其中,R 1a、R 1b和R 1c独立地为C 6-C 18芳基、被R 1a-1取代的C 6-C 18芳基或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 1a-1独立地为为羟基、卤素或C 1-C 12烷基;R 2a、R 4a、R 2b、R 4b、R 2c和R 5c独立地为H或卤素;R 3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 4c为H、3-6元的杂环烷基或被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基,所述的3-6元的杂环烷基中的杂原子和被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多 种,杂原子个数为1-3个;R 4c-1和R 4c-2独立地为C 1-C 4烷基;A a和A b独立地为CR 5a或N;R 5a为H、C 1-C 4烷基或卤素;n1为1、2或3;M 1为3-9元杂环烷基、被M 1-1取代的3-9元杂环烷基、-3-9元杂环烷基-(CH 2) n1’OC(=O)-或不存在;所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH 2) n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个;n1’为1、2或3;M 1-1为C 1-C 4烷基;Y 1为O、-NR 8或不存在(即M 1与L a直接相连);环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;n2为0、1、2或3;Y 2为O、-NR 9或不存在(即X a中的亚烷基与L a直接相连);R 8和R 9独立地为C 1-C 4烷基;n3为1、2或3;n3’为0、1、2或3;R 10和R 11独立地为C 1-C 4烷基;环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;L a、L b和L c为连接接团;Q a、Q b和Q c为E3泛素连接酶配体;方案3:所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物:其中,R 1a、R 1b和R 1c独立地为C 6-C 18芳基、被R 1a-1取代的C 6-C 18芳基或5-10杂芳基,所述的5-10杂芳基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 1a-1为羟基、卤素或C 1-C 12烷基;R 2a、R 4a、R 2b、R 4b、R 2c和R 5c独立地为H或卤素;R 3a为7-12元的桥环杂环烷基,所述的桥环杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-4个;R 4c为H、3-6元的杂环烷基或被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基,所述的3-6元的杂环烷基中的杂原子和被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个;R 4c-1和R 4c-2独立地为C 1-C 4烷基;A a和A b独立地为CR 5a或N;R 5a为H、C 1-C 4烷基或卤素;n1为1、2或3;M 1为3-9元杂环烷基、被M 1-1取代的3-9元杂环烷基、-3-9元杂环烷基-(CH 2) n1’OC(=O)-或不存在;所述的3-9元杂环烷基和所述的3-9元杂环烷基-(CH 2) n1’中的杂原子独立地为N、O或S中的一种或多种,杂原子个数为1-3个;n1’为1、2或3;M 1-1为C 1-C 4烷基;Y 1为O、-NR 8或不存在;环D为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;n2为0、1、2或3;Y 2为O、-NR 9或不存在;R 8和R 9独立地为C 1-C 4烷基;n3为1、2或3;n3’为1、2或3;R 10和R 11独立地为C 1-C 4烷基;环E为3-6元的杂环烷基,所述的杂环烷基中的杂原子为N,个数为1或2个;L a、L b和L c为连接接团;Q a、Q b和Q c为E3泛素连接酶配体;方案4:所述的如式Ia所示的化合物的通式为式Ia-1:其中,R 2a为卤素;R 3a为7-12元的桥环杂环烷基,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子为N,个数为2个,R 5a独立地为卤素;M 1为-3-9元杂环烷基-(CH 2) n1OC(=O)-或不存在,Y 1为-NR 8;或者,M 1为3-9元杂环烷基或被M 1-1取代的3-9元杂环烷基,Y 1为O或不存在;方案5:所述的如式Ia所示的化合物的通式为式Ia-1:其中,R 2a为卤素;R 3a为7-12元的桥环杂环烷基,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子为N,个数为2个,R 5a为卤素;方案6:所述的如式Ia所示的化合物通式为式Ia-2:其中,R 2a为卤素;R 5a为卤素;方案7:所述的如式Ia所示的化合物的通式为式Ia-3:其中,R 2a为卤素;R 3a为7-12元的桥环杂环烷基,所述的7-12元的桥环杂环烷基为7或8元桥环杂环烷基,杂原子为N,个数为2个,R 5a为卤素;M 1为3-9元杂环烷基或被M 1-1取代的3-9元杂环烷基;Y 1为不存在;方案8:所述的如式Ib所示的化合物的通式为式Ib-1:其中,R 5a为卤素;R 2b为卤素;方案9:所述的如式Ib所示的化合物的通式为式Ib-1:其中,R 5a为卤素;R 2b为卤素;n3’为0;n7为0,n8为3、4或5,Y 3为CH 2或不存在;n10和n13为0,n11和n12独立地为3、4或5,Y 4为不存在;方案10:所述的如式Ic所示的化合物的通式为式Ic'或Ic”:优选,所述的式Ic'或Ic”中:R 2a和R 5c独立地为卤素;n4和n6为2,n5为4;R 4c为被-N(R 4c-1R 4c-2)取代的3-6元的杂环烷基。
- 如权利要求1所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物,其特征在于,所述的如式Ia、 Ib或Ic所示的化合物药学上可接受的盐的盐型为甲酸盐、三氟醋酸盐或盐酸盐;和/或,所述的如式Ia、Ib或Ic所示的化合物药学上可接受的盐中盐的个数为1个、2个、3个、4个、5个或6个。
- 一种药物组合物,其包括物质X和一种或多种药用辅料,所述的物质X为如权利要求1-11中任一项所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物。
- 一种物质Y在制备用于治疗或预防与KRAS突变所介导的癌症的药物中的应用;所述的物质Y为如权利要求1-11中任一项所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其前药、其代谢产物或其同位素化合物或如权利要求13所述的药物组合物。
- 一种物质Y在制备用于治疗或预防癌症的药物中的应用;所述的物质Y为如权利要求1-11中任一项所述的如式Ia、Ib或Ic所示的化合物、其药学上可接受的盐、其溶剂合物、其立体异构体、其互变异构体、其前药、其代谢产物或其同位素化合物或如权利要求13所述的药物组合物;所述的癌症为血液癌症、胰腺癌、MYH相关性息肉病、结肠直肠癌或肺癌。
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Cited By (3)
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WO2023081476A1 (en) * | 2021-11-05 | 2023-05-11 | Ranok Therapeutics (Hangzhou) Co. Ltd. | Methods and compositions for targeted protein degradation |
WO2023138524A1 (zh) * | 2022-01-24 | 2023-07-27 | 贝达药业股份有限公司 | Kras g12d降解剂及其在医药上的应用 |
WO2023215906A1 (en) * | 2022-05-06 | 2023-11-09 | Hangzhou Jijing Pharmaceuticaltechnology Limited | Kras g12d proteolysis targeting chimeras |
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