WO2020074004A1 - 环二核苷酸类化合物及其应用 - Google Patents
环二核苷酸类化合物及其应用 Download PDFInfo
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- WO2020074004A1 WO2020074004A1 PCT/CN2019/110800 CN2019110800W WO2020074004A1 WO 2020074004 A1 WO2020074004 A1 WO 2020074004A1 CN 2019110800 W CN2019110800 W CN 2019110800W WO 2020074004 A1 WO2020074004 A1 WO 2020074004A1
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- 0 Nc1ncnc2c1nc[n]2[C@@](C1F)O[C@](CO[P@](OC(C2F)[C@@](CO3)O[C@]2[n]2c(N=C(N)NC4=O)c4nc2)(S)=O)C1O*3=O Chemical compound Nc1ncnc2c1nc[n]2[C@@](C1F)O[C@](CO[P@](OC(C2F)[C@@](CO3)O[C@]2[n]2c(N=C(N)NC4=O)c4nc2)(S)=O)C1O*3=O 0.000 description 134
- SERYYMHERVXUTA-OSIBBQKWSA-N CC([C@@H](C1F)O[C@H](COCc2ccccc2)C1OCc1ccccc1)=O Chemical compound CC([C@@H](C1F)O[C@H](COCc2ccccc2)C1OCc1ccccc1)=O SERYYMHERVXUTA-OSIBBQKWSA-N 0.000 description 1
- KJTMBNPILUYPGZ-UHFFFAOYSA-N COCCC(C12)C1(O)OC(COCc1ccccc1)=C2OCc1ccccc1 Chemical compound COCCC(C12)C1(O)OC(COCc1ccccc1)=C2OCc1ccccc1 KJTMBNPILUYPGZ-UHFFFAOYSA-N 0.000 description 1
- XKSFLKWHSNPULH-YNIZLOQISA-N CO[C@@H](C1F)O[C@H](COCc2ccccc2)C1OCc1ccccc1 Chemical compound CO[C@@H](C1F)O[C@H](COCc2ccccc2)C1OCc1ccccc1 XKSFLKWHSNPULH-YNIZLOQISA-N 0.000 description 1
- VTHYHRRWPVMJCI-URYLQHRCSA-N Nc1ncnc2c1nc[n]2[C@@H](C1F)OC(CO)=C1O Chemical compound Nc1ncnc2c1nc[n]2[C@@H](C1F)OC(CO)=C1O VTHYHRRWPVMJCI-URYLQHRCSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H23/00—Compounds containing boron, silicon or a metal, e.g. chelates or vitamin B12
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
- C07H19/213—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids containing cyclic phosphate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
Definitions
- the present invention relates to compounds represented by formula (I), their optical isomers and their pharmaceutically acceptable salts, and the use of the compounds as STING agonists.
- STING is activated in three ways: 1) Exogenous (3 ', 3') cyclic dinucleotides (c-diGMP, c-diAMP, and c-GAMP) released by binding invading bacteria or archaea ) Activation, which shows that STING has the role of innate immune activation in anti-infection; 2) activation by binding (2'3 ') cyclic guanosine monophosphate adenosine monophosphate (2', 3'c-GAMP) , Which is induced by circular GMP-AMP dinucleotide synthetase (cGAS) in the presence of foreign double-stranded DNA (eg, released by invading bacteria, viruses, or protozoa) or self DNA in mammals
- cGAS circular GMP-AMP dinucleotide synthetase
- Source circular dinucleotide which shows that STING has the effect of innate immunity induced by endogenous or exogenous DNA
- STING acts as a receptor for DNA in the cytoplasm. Its activation can lead to the activation of two downstream pathways, IRF3 and NF- ⁇ B, to activate the immune system. Activation of the NF- ⁇ B pathway leads to the activation of a series of proinflammatory cytokines downstream, while activation of the IRF3 pathway leads to the activation of type I interferon (IFN- ⁇ / ⁇ ), dendritic cells, cytotoxic cells, NK cells, etc. Activation, thereby exerting an anti-tumor effect.
- IFN- ⁇ / ⁇ type I interferon
- the DNA in the human body usually does not activate the STING protein, because under normal circumstances DNA can only exist in the nucleus (except mitochondrial DNA). But if DNA leaks into the cytoplasm, it will activate STING and trigger an immune response. Recently, it was found that radiotherapy and chemotherapy can also activate STING, which may also be caused by DNA leakage in dead tumor cells.
- the present invention provides the compound represented by formula (I), its optical isomer and its pharmaceutically acceptable salt,
- R 1 and R 1a are independently selected from
- T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , T 13 are each independently selected from -C (R)-and -N-;
- R is independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino, of which C 1-6 alkyl, C 1-6 alkoxy, C 1-6 Alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R's;
- R ' is selected from F, Cl, Br, I, OH, NH 2 and CH 3 ;
- R 2 and R 2a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino And C 2-6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl are optionally substituted by 1, 2 or 3 R substitutions;
- R 3 and R 3a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino And C 2-6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl are optionally substituted by 1, 2 or 3 R substitutions;
- R 4 and R 4a are independently selected from H, halogen, OH, NH 2 , CN, N 3 , C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1- 6 alkylamino and C 2-6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl are optional Replaced by 1, 2 or 3 R;
- R 5 and R 5a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino And C 2-6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl are optionally substituted by 1, 2 or 3 R substitutions;
- R 6 and R 6a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino And C 2-6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R;
- R 7 and R 7a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino , Wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R;
- R 10 and R 10a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino , Wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R;
- R 7 and R 10 are joined together to form a C 3-6 cycloalkyl, C 3-6 cycloalkenyl, or C 3-6 cycloalkynyl, the C 3-6 cycloalkyl, C 3- 6- ring alkenyl or C 3-6 cycloalkynyl is optionally substituted with 1, 2 or 3 R;
- R 7a and R 10a are connected together to form a C 3-6 cycloalkyl, C 3-6 cycloalkenyl or C 3-6 cycloalkynyl, the C 3-6 cycloalkyl, C 3-6 ring Alkenyl or C 3-6 cycloalkynyl is optionally substituted with 1, 2 or 3 R;
- R 8 is selected from BH 3 - and -S (R 9 );
- R 9 is selected from H, CH 2 OC ( ⁇ O) R 11 , CH 2 OC ( ⁇ O) OR 11 , CH 2 CH 2 SC ( ⁇ O) R 11 and CH 2 CH 2 SSCH 2 R 11 ;
- R 11 is selected from C 6-10 aryl, 5-10 membered heteroaryl, C 1-6 heterocycloalkyl and C 1 - 20 alkyl group, a C 1 - 20 alkyl optionally substituted with 1,2, 3. 4 or 5 C 6-10 aryl, C 3-10 cycloalkyl, OH and F substitutions;
- R 4 and R 6 or R 4a and R 6a are joined together to form a 5-6 membered heterocycloalkyl
- X 1 and X 1a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 2 and X 2a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 3 and X 3a are independently selected from -O- and -S-;
- R 8 when the above-described R 8 is selected from BH 3 - when, R 4 a and wherein R 4a is selected from F, Cl and Br, the other is selected from F, Cl, Br, OH, OCH 3 or N 3 ,
- R 4a is selected from F, Cl and Br
- R 4a is selected from F, Cl and Br
- the other is selected from F, Cl, Br, OH, OCH 3 or N 3
- Other variables are as defined in the present invention.
- the above R is independently selected from H, halogen, OH, NH 2 , CN, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 alkylthio and C 1-3 alkylamino, of which C 1-3 alkyl, C 1-3 alkoxy, C 1-3
- the alkylthio group and the C 1-3 alkylamino group are optionally substituted with 1, 2, or 3 R ′, and other variables are as defined in the present invention.
- R is independently selected from H, F, Cl, Br, I, OH, NH 2 , CN, Me, Where Me, It can be optionally substituted with 1, 2 or 3 R ', and other variables are as defined in the present invention.
- the above R is independently selected from H, F, Cl, Br, I, OH, NH 2 , CN, Me, Other variables are as defined in the present invention.
- R 1 and R 1a are independently selected from Other variables are as defined in the present invention.
- R 1 and R 1a are independently selected from Other variables are as defined in the present invention.
- R 2 , R 2a , R 3 , R 3a , R 5 and R 5a , R 6 and R 6a are independently selected from H, and other variables are as defined in the present invention.
- R 6 and R 6a are independently selected from H and methyl, and other variables are as defined in the present invention.
- R 4 and R 4a are independently selected from F, OH, NH 2 , N 3 and Other variables are as defined in the present invention.
- R 7 and R 7a are independently selected from H and CH 3 , and other variables are as defined in the present invention.
- the above compound, its optical isomer and its pharmaceutically acceptable salt are selected from
- R 1 and R 1a are independently selected from
- R 4 and R 4a are independently selected from H, halogen, OH, NH 2 , CN, N 3 , C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1- 6 alkylamino and C 2-6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl are optional Replaced by 1, 2 or 3 R;
- R 6 is selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkyne Group, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R;
- R 4 and R 6 are joined together to form a 5-6 membered heterocycloalkyl
- R 8 is selected from BH 3 - and -S (R 9 );
- R 9 is selected from H, CH 2 OC ( ⁇ O) R 11 , CH 2 OC ( ⁇ O) OR 11 , CH 2 CH 2 SC ( ⁇ O) R 11 and CH 2 CH 2 SSCH 2 R 11 ;
- R 11 is selected from C 6-10 aryl, 5-10 membered heteroaryl, C 1-6 heterocycloalkyl and C 1 - 20 alkyl group, a C 1 - 20 alkyl optionally substituted with 1,2, 3. 4 or 5 C 6-10 aryl, C 3-10 cycloalkyl, OH and F substitutions;
- X 1 and X 1a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 2 and X 2a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 3 and X 3a are independently selected from -O- and -S-;
- R 8 is selected from BH 3 - when, R 4 a and wherein R 4a is selected from F, Cl and Br, the other is selected from F, Cl, Br, OH, OCH 3 or N 3;
- R 8 is selected from -S (R 9 ), one of R 4 and R 4a is selected from F, Cl and Br, and the other is selected from OH, OCH 3 or N 3 ;
- R 8 is selected from -S (R 9 ) and one of R 4 and R 4a is selected from F, Cl and Br, and the other is not selected from OH, OCH 3 or N 3 , R 4 and R 4a are not Select from And R 4 and R 4a are not simultaneously selected from
- the above-mentioned compound, its optical isomer and its pharmaceutically acceptable salt are selected from
- R 1 , R 1a , R 4a , R 7 , R 7a , R 8 and R 6a are as defined above.
- the above-mentioned compound, its optical isomer and its pharmaceutically acceptable salt are selected from
- the above compound, its optical isomer and its pharmaceutically acceptable salt are selected from
- R 1 and R 1a are independently selected from
- X 1 and X 1a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 2 and X 2a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 3 and X 3a are independently selected from -O- and -S-;
- the above compound, its optical isomer and its pharmaceutically acceptable salt are selected from
- R 1 and R 1a are independently selected from Other variables are as defined in the present invention.
- the present invention provides the compound represented by formula (I), its optical isomer and its pharmaceutically acceptable salt,
- R 1 and R 1a are independently selected from
- T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , T 13 are each independently selected from -C (R)-and -N-;
- R is independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino, of which C 1-6 alkyl, C 1-6 alkoxy, C 1-6 Alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R's;
- R ' is selected from F, Cl, Br, I, OH, NH 2 and CH 3 ;
- R 2 and R 2a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2- 6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl group optionally substituted by 1, 2 or 3 R substitutions;
- R 3 and R 3a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2- 6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl group optionally substituted by 1, 2 or 3 R substitutions;
- R 4 and R 4a are independently selected from H, halogen, OH, NH 2 , CN, N 3 , C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1- 6 alkylamino and C 2-6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl are optional Replaced by 1, 2 or 3 R;
- R 5 and R 5a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2- 6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2-6 alkynyl group optionally substituted by 1, 2 or 3 R substitutions;
- R 6 and R 6a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino and C 2- 6 alkynyl, wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, and C 1-6 alkylamino optionally substituted with 1, 2 or 3 R <
- R 7 and R 7a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino , Wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R;
- R 10 and R 10a are independently selected from H, halogen, OH, NH 2 , CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino , Wherein C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio and C 1-6 alkylamino are optionally substituted with 1, 2 or 3 R;
- R 7 and R 10 are joined together to form a C 3-6 cycloalkyl, C 3-6 cycloalkenyl, or C 3-6 cycloalkynyl, the C 3-6 cycloalkyl, C 3- 6- ring alkenyl or C 3-6 cycloalkynyl is optionally substituted with 1, 2 or 3 R;
- R 7a and R 10a are connected together to form a C 3-6 cycloalkyl, C 3-6 cycloalkenyl or C 3-6 cycloalkynyl, the C 3-6 cycloalkyl, C 3-6 ring Alkenyl or C 3-6 cycloalkynyl is optionally substituted with 1, 2 or 3 R;
- R 8 is selected from BH 3 - and -S (R 9 );
- R 9 is selected from H, CH 2 OC ( ⁇ O) R 11 , CH 2 OC ( ⁇ O) OR 11 , CH 2 CH 2 SC ( ⁇ O) R 11 and CH 2 CH 2 SSCH 2 R 11 ;
- R 11 is selected from C 6-10 aryl, 5-10 membered heteroaryl, C 1-6 heterocycloalkyl and C 1 - 20 alkyl group, a C 1 - 20 alkyl optionally substituted with 1,2, 3. 4 or 5 C 6-10 aryl, C 3-10 cycloalkyl, OH and F substitutions;
- R 4 and R 6 or R 4a and R 6a are joined together to form a 5-6 membered heterocycloalkyl
- X 1 and X 1a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 2 and X 2a are independently selected from -NH-, -O-, -S- and -CH 2- ;
- X 3 and X 3a are independently selected from -O- and -S-;
- the above R is independently selected from H, halogen, OH, NH 2 , CN, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 alkylthio and C 1-3 alkylamino, of which C 1-3 alkyl, C 1-3 alkoxy, C 1-3
- the alkylthio and C 1-3 alkylamino are optionally substituted with 1, 2, or 3 R '.
- R is independently selected from H, F, Cl, Br, I, OH, NH 2 , CN, Me, Where Me, It is optionally substituted with 1, 2 or 3 R '.
- the above R is independently selected from H, F, Cl, Br, I, OH, NH 2 , CN, Me,
- R 1 and R 1a are independently selected from
- R 1 and R 1a are independently selected from
- R 2 , R 2a , R 3 , R 3a , R 5 , R 5a , R 6 and R 6a are each independently selected from H.
- R 4 and R 4a are independently selected from F, OH, NH 2 , N 3 and
- R 7 and R 7a are independently selected from H and CH 3 .
- the above-mentioned compound, its optical isomer and its pharmaceutically acceptable salt are selected from
- R 1 , R 1a , R 4a , R 7 , R 7a , R 8 and R 6a are as defined above.
- the present invention also provides a compound of the formula, its optical isomer and its pharmaceutically acceptable salt, which is selected from
- pharmaceutically acceptable refers to those compounds, materials, compositions and / or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit / risk ratio.
- pharmaceutically acceptable salt refers to a salt of a compound of the present invention, prepared from a compound having a specific substituent and a relatively non-toxic acid or base found in the present invention.
- base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent.
- Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts.
- acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent.
- Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Bisulfate, hydroiodic acid, phosphorous acid, etc .; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also includes salts of amino acids (such as arginine, etc.) , And salts of organic acids such as glucuronic acid. Certain compounds of the present invention contain basic and acidic functional groups and can be converted to any base or
- the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of appropriate base or acid in water or an organic solvent or a mixture of both.
- the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
- the present invention contemplates all such compounds, including cis and trans isomers, (-)-and (+)-enantiomers, (R)-and (S) -enantiomers, diastereomers Isomers, (D) -isomers, (L) -isomers, and their racemic mixtures and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this Within the scope of the invention. Additional asymmetric carbon atoms may be present in the substituents such as alkyl. All these isomers and their mixtures are included in the scope of the present invention.
- wedge-shaped solid line key And wedge-shaped dotted keys Represents the absolute configuration of a three-dimensional center
- using straight solid line keys And straight dotted keys Represents the relative configuration of the three-dimensional center
- wavy lines Represents a wedge-shaped solid line key Or wedge-shaped dotted key Or with wavy lines Represents a straight solid line key And straight dotted keys
- tautomer or “tautomeric form” means that at room temperature, isomers of different functional groups are in dynamic equilibrium and can quickly convert to each other. If tautomers are possible (as in solution), the chemical equilibrium of tautomers can be achieved.
- proton tautomers also known as prototropic tautomers
- proton tautomers include interconversion through proton migration, such as keto-enol isomerization and imine-ene Amine isomerization.
- Valence tautomer (valence tautomer) includes some recombination of bond-forming electrons to perform mutual conversion.
- keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one, or E.g Tautomers.
- the terms “rich in one isomer”, “isomer enriched”, “rich in one enantiomer” or “enantiomerically enriched” refer to one of the isomers or pairs
- the content of the enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or 96% or greater, or 97% or greater, or 98% or greater, or 99% or greater, or 99.5% or greater, or 99.6% or greater, or 99.7% or greater, or 99.8% or greater, or greater or equal 99.9%.
- the terms “isomer excess” or “enantiomeric excess” refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the excess of isomer or enantiomer (ee value) is 80% .
- optically active (R)-and (S) -isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, in which the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure The desired enantiomer.
- a diastereomeric salt is formed with an appropriate optically active acid or base, and then by conventional methods known in the art The diastereomers are resolved and the pure enantiomers are recovered.
- the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography, which uses a chiral stationary phase, and is optionally combined with chemical derivatization methods (eg, amino groups are generated from amine Formate).
- the compound of the present invention may contain unnatural proportions of atomic isotopes in one or more atoms constituting the compound.
- compounds can be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C).
- the hydrogen can be replaced by heavy hydrogen to form a deuterated drug.
- the bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon.
- deuterated drugs have lower toxicity and increase drug stability , Strengthen the efficacy, extend the biological half-life of drugs and other advantages.
- the conversion of all isotopic compositions of the compounds of the present invention, whether radioactive or not, is included within the scope of the present invention.
- “Optional” or “optionally” means that the subsequently described event or condition may, but need not necessarily occur, and that the description includes situations where the event or condition occurs and circumstances where the event or condition does not occur.
- substituted means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which may include heavy hydrogen and hydrogen variants, as long as the valence state of the specific atom is normal and the substituted compound is stable of.
- Oxygen substitution does not occur on aromatic groups.
- optionally substituted means that it may or may not be substituted. Unless otherwise specified, the type and number of substituents may be arbitrary on the basis of chemical realization.
- any variable (such as R) appears more than once in the composition or structure of a compound, its definition in each case is independent.
- R when any variable (such as R) appears more than once in the composition or structure of a compound, its definition in each case is independent.
- the group can optionally be substituted with up to two Rs, and R in each case has independent options.
- combinations of substituents and / or variants thereof are only allowed if such combinations will produce stable compounds.
- connection direction is arbitrary, for example,
- the linking group L in the middle is -MW-, then -MW- can be formed by connecting the benzene ring and cyclopentane in the same direction as the reading order from left to right It can also be formed by connecting the benzene ring and cyclopentane in the opposite direction to the reading order from left to right
- Combinations of the linking group, substituents, and / or variants thereof are only allowed if such a combination will produce a stable compound.
- the number of atoms on a ring is usually defined as the number of members of the ring.
- “5-6 membered ring” refers to a “ring” with 5-6 atoms arranged around it.
- 5-6 membered ring means cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl consisting of 5 to 6 ring atoms Radical or heteroaryl.
- the ring includes a single ring, and also includes a double ring system such as a spiro ring, a bicyclic ring, and a bridge ring. Unless otherwise specified, the ring optionally contains 1, 2 or 3 heteroatoms independently selected from O, S and N.
- the 5-6 member ring includes 5 member, 6 member ring, and the like.
- 5-6 membered ring includes, for example, phenyl, pyridyl, piperidinyl, and the like; on the other hand, the term “5-6 membered heterocycloalkyl” includes piperidinyl and the like, but does not include phenyl.
- ring also includes ring systems containing at least one ring, where each "ring” independently conforms to the above definition.
- C 1-20 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 20 carbon atoms.
- the C 1-20 alkyl group includes C 1-10 , C 1-9 , C 1-8 , C 1-6 , C 1-5 , C 1-14 , C 1-3 , C 1-2 , C 2-16 , C 2-4 , C 10 , C 8 , C 7 , C 6 and C 5 alkyl, etc .; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent ( Such as methine).
- C 1-20 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, heptyl, octyl and so on.
- Me methyl
- Et ethyl
- propyl including n-propyl and isopropyl
- butyl including n-butyl, isobutyl , S-butyl and t-butyl
- pentyl including n-pentyl, isopentyl and neopentyl
- hexyl heptyl, octyl and so on.
- C 1-6 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 6 carbon atoms.
- the C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 and C 5 alkyl groups; etc .; Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine).
- C 1-6 alkyl examples include but are not limited to methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl and so on.
- C 1-3 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 3 carbon atoms.
- the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc .; it may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine) .
- Example C 1- 3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n- propyl and isopropyl) and the like.
- heteroalkyl by itself or in combination with another term means a stable linear or branched alkyl radical consisting of a certain number of carbon atoms and at least one heteroatom or heteroatom group or a combination thereof.
- the heteroatom is selected from B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized.
- the heteroalkyl is C 1-6 heteroalkyl; in other embodiments, the heteroalkyl is C 1-3 heteroalkyl.
- the heteroatom or heteroatom group may be located at any internal position of the heteroalkyl group, including the attachment position of the alkyl group to the rest of the molecule, but the terms "alkoxy”, “alkylamino” and “alkylthio” (or thioalkane Oxygen) is a conventional expression and refers to those alkyl groups that are connected to the rest of the molecule through an oxygen atom, an amino group, or a sulfur atom, respectively.
- C 1-6 alkoxy refers to those alkyl groups containing 1 to 6 carbon atoms connected to the rest of the molecule through one oxygen atom.
- the C 1-6 alkoxy group includes C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 , C 5 , C 4 and C 3 alkoxy groups, etc. .
- C 1-6 alkoxy groups include but are not limited to methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy Oxy, s-butoxy and t-butoxy), pentyloxy (including n-pentyloxy, isopentyloxy and neopentyloxy), hexyloxy, etc.
- C 1-3 alkoxy refers to those alkyl groups containing 1 to 3 carbon atoms connected to the rest of the molecule by one oxygen atom.
- the C 1-3 alkoxy group includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy groups and the like.
- Examples of C 1-3 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
- C 1-6 alkylamino refers to those alkyl groups containing 1 to 6 carbon atoms attached to the rest of the molecule through an amino group.
- the C 1-6 alkylamino group includes C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 , C 5 , C 4 , C 3 and C 2 alkyl amino groups Wait.
- C 1-6 alkylamino examples include but are not limited to -NHCH 3 , -N (CH 3 ) 2 , -NHCH 2 CH 3 , -N (CH 3 ) CH 2 CH 3 , -N (CH 2 CH 3 ) ( CH 2 CH 3 ), -NHCH 2 CH 2 CH 3 , -NHCH 2 (CH 3 ) 2 , -NHCH 2 CH 2 CH 2 CH 3, etc.
- C 1-3 alkylamino refers to those alkyl groups containing 1 to 3 carbon atoms attached to the rest of the molecule through an amino group.
- the C 1-3 alkylamino group includes C 1-2 , C 3 and C 2 alkylamino groups and the like.
- Examples of C 1-3 alkylamino groups include, but are not limited to, -NHCH 3 , -N (CH 3 ) 2 , -NHCH 2 CH 3 , -N (CH 3 ) CH 2 CH 3 , -NHCH 2 CH 2 CH 3 ,- NHCH 2 (CH 3 ) 2 etc.
- C 1-6 alkylthio refers to those alkyl groups containing 1 to 6 carbon atoms connected to the rest of the molecule through a sulfur atom.
- the C 1-6 alkylthio group includes C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 , C 5 , C 4 , C 3 and C 2 alkane Sulfur-based.
- Examples of C 1-6 alkylthio groups include, but are not limited to, -SCH 3 , -SCH 2 CH 3 , -SCH 2 CH 2 CH 3 , -SCH 2 (CH 3 ) 2 and the like.
- C 1-3 alkylthio refers to those alkyl groups containing 1 to 3 carbon atoms connected to the rest of the molecule through a sulfur atom.
- the C 1-3 alkylthio group includes C 1-3 , C 1-2 and C 3 alkylthio groups.
- Examples of C 1-3 alkylthio groups include, but are not limited to, -SCH 3 , -SCH 2 CH 3 , -SCH 2 CH 2 CH 3 , -SCH 2 (CH 3 ) 2 and the like.
- C 2-6 alkynyl is used to indicate a straight-chain or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon triple bond, carbon-carbon triple bond It can be located anywhere on the group.
- the C 2-6 alkynyl group includes C 2-4 , C 2-3 , C 4 , C 3 and C 2 alkynyl groups. It can be monovalent, bivalent or multivalent. Examples of C 2-6 alkynyl include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and the like.
- the term "5-6 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 5 to 6 ring atoms, with 1, 2, 3 or 4 ring atoms Are heteroatoms independently selected from O, S, and N, and the rest are carbon atoms, wherein nitrogen atoms are optionally quaternized, and nitrogen and sulfur heteroatoms may be optionally oxidized (ie, NO and S (O) p , p Is 1 or 2). It includes single-ring and double-ring systems, wherein the double-ring system includes spiro ring, parallel ring and bridge ring.
- the hetero atom may occupy the connection position of the heterocyclic alkyl group to the rest of the molecule.
- the 5-6 membered heterocycloalkyl group includes 5-membered and 6-membered heterocycloalkyl groups.
- 5-6 membered heterocycloalkyl examples include, but are not limited to, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothien-2-yl and tetrahydrothien-3-yl, etc.) , Tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, etc.), piperazinyl (including 1 -Piperazinyl and 2-piperazinyl etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazole Alkyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydro
- C 6-10 aromatic ring and “C 6-10 aryl group” of the present invention can be used interchangeably.
- the term “C 6-10 aromatic ring” or “C 6-10 aryl group” means A cyclic hydrocarbon group consisting of 6 to 10 carbon atoms with a conjugated ⁇ -electron system. It can be a single ring, a fused bicyclic ring, or a fused tricyclic ring system, where each ring is aromatic. It may be monovalent, divalent or multivalent, and C 6-10 aryl groups include C 6-9 , C 9 , C 10 and C 6 aryl groups and the like. Examples of C 6-10 aryl groups include, but are not limited to, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl, etc.).
- 5-10 membered heteroaryl ring and “5-10 membered heteroaryl group” of the present invention can be used interchangeably.
- the term “5-10 membered heteroaryl group” means from 5 to 10 rings The cyclic group consisting of atoms with a conjugated ⁇ electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. It can be a monocyclic ring, fused bicyclic ring or fused tricyclic ring system, where each ring is aromatic.
- nitrogen and sulfur heteroatoms can be optionally oxidized (ie NO and S (O) p , p is 1 or 2).
- the 5-10 membered heteroaryl group can be attached to the rest of the molecule through a heteroatom or carbon atom.
- the 5-10 membered heteroaryl group includes 5-8 membered, 5-7 membered, 5-6 membered, 5 membered, and 6 membered heteroaryl groups.
- Examples of the 5-10 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl, and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyryl Oxazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5- Oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1, 2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, etc.), thiazolyl (including 2-thiazoly
- C n-n + m or C n -C n + m includes any specific case of n to n + m carbons, for example, C 1-6 includes C 1 , C 2 , C 3 , C 4 , C 5 and C 6 , also including any range from n to n + m, for example, C 1-6 includes C 1-3 , C 1-6 , C 1-4 , C 3-6 , C 3- 5 , C 2-5 and C 1- 5 etc .; in the same way, n to n + m means that the number of atoms in the ring is n to n + m, for example, 5-6 member ring includes 5 member ring and 6 member ring .
- the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalently, preferred embodiments include but are not limited to the embodiments of the present invention.
- HPLC detection conditions of the present invention are as follows: chromatographic column: YMC-Pack ODS-A 150 * 4.6mm, 5 ⁇ m, mobile phase: water (0.06875% trifluoroacetic acid) -acetonitrile (0.0625% trifluoroacetic acid); flow rate: 1.0mL / min; detection wavelength: UV 220nm & 215nm &254nm; column temperature: 40 °C.
- FIG. 1 Experimental results of 4T1 breast cancer homologous mouse model
- FIG. 1 CT-26 colon cancer homologous mouse model drug efficacy test results
- Figure 3 MC38 colon cancer homologous mouse model drug efficacy test results.
- trimethylchlorosilane (1.61g, 14.86mmol, 1.89mL) was added dropwise to the solution of compound 1-1 (1g, 3.71mmol) in pyridine (20mL).
- the benzoyl chloride (605mg, 4.30mmol, 500.00 ⁇ L) was added to the reaction system, and the temperature was raised to 15 ° C for 3h.
- compound 1-8 (15g, 22.26mmol) and imidazole (4.55g, 66.79mmol) were dissolved in pyridine (60mL), and tert-butyldimethylchlorosilane (5.03g, 33.40mmol, 4.09) was added mL), the reaction system was stirred at room temperature for 4h. Ethyl acetate (200mL) was added to the system for dilution, the white solid was filtered off, the solution was concentrated to dryness, redissolved in ethyl acetate (200mL), washed with saturated brine (100mL x 4), dried over anhydrous sodium sulfate, and filtered.
- Step 13 Preparation of compounds 1-14A, 1-14B, 1-14C and 1-14D
- the crude compound 1-14 was dissolved in water (10mL) and efficiently prepared for liquid phase separation (separation conditions: chromatographic column: Xbridge 150 * 30mm * 10 ⁇ m; mobile phase: [water (10mM ammonium bicarbonate) -acetonitrile]; acetonitrile%: 12% -32%, flow rate: 25mL / min, 7min).
- the photoactive isomer compound 1-14A (20 mg, 24.31 ⁇ mol) was dissolved in pyridine (2 mL), and triethylamine (290.80 mg, 2.87 mmol, 0.4 mL) and triethylamine trihydrofluoride (197.80) were added sequentially. mg, 1.23mmol, 0.2mL), the reaction system was heated to 50 °C and stirred for 14h. After cooling to room temperature, isopropyloxytrimethylsilane (745 mg, 5.63 mmol, 1 mL) was added, and the reaction was continued at room temperature for 4 h.
- optically active pure isomers 1B, 1C, and 1D can be prepared from compounds 1-14B, 1-14C, and 1-14D, respectively, referring to the preparation method of compound 1A.
- the compound 1-5 (1.8g, 2.66mmol), Molecular sieve (2g) and tetrazole (0.45M acetonitrile solution, 88.80mL) were dispersed in acetonitrile (10mL).
- acetonitrile 10mL
- a solution of compound 2-1 (2.33 g, 2.66 mmol) in acetonitrile (10 mL) was added.
- the reaction was stirred at room temperature for 1 hour.
- the reaction solution was diluted with ethyl acetate (50 mL) and filtered. The filtrate was washed with saturated sodium bicarbonate solution (40 mL x 3) and saturated brine (10 mL), dried over anhydrous magnesium sulfate, and concentrated in vacuo to give the compound 2-2.
- BH 3 -Me 2 S (2M in tetrahydrofuran, 3.31 mL) was slowly added to compound 2-2 (3.2 g, 2.21 mmol), In a mixed solution of molecular sieve (3g) and dichloromethane (35mL). The reaction was stirred at room temperature for 40 min and then filtered. The filter cake was washed with ethyl acetate (50 mL). Water (20 mL) was added to the filtrate, and then extracted with ethyl acetate (20 mL x 3). The combined organic phase was dried over Na 2 SO 4 and concentrated under reduced pressure to obtain compound 2-3, and the crude product was directly used in the next reaction.
- compound 3-3 (100 mg, 0.34 mmol) was dissolved in acetonitrile (10 mL), and sodium iodide (250 mg, 1.68 mmol) and trimethyliodosilane (0.2 mL, 1.56 mmol) were added in sequence.
- compound 3-5 (0.85g, 1.45mmol) was dissolved in pyridine (8mL), followed by imidazole (200mg, 2.94mmol) and tert-butyldimethylchlorosilane (265mg, 1.76mmol), the reaction system The reaction was stirred at 25 ° C for 12h.
- Step 10 Preparation of compounds 3-12A, 3-12B, 3-12C, 3-12D
- compound 5-1 (20 g, 76.84 mmol) was dissolved in acetonitrile (300 mL), sodium hydride (4.61 g, 115.26 mmol, 60%) was added, and after stirring for 0.5 h, benzyl bromide (13.14 g, 76.84mmol), warmed to 20 °C stirring reaction for 3h.
- compound 5-4 (17.94g, 64.46mmol) was dissolved in dioxane (45mL) and water (40mL), formalin (36mL, 483.54mmol, 37% aqueous solution) and hydrogen were added Sodium oxide (1M in water, 176mL), the reaction was warmed to 20 ° C and stirred for 48h.
- the reaction solution was extracted with ethyl acetate (200 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, the solid was filtered off, and the filtrate was concentrated to obtain the crude product 5-5, which was directly used in the next reaction without further purification.
- compound 5-12 (1g, 3.58mmol) was dissolved in pyridine (20mL) and N, N-dimethylformamide (10mL), and trimethylchlorosilane (1.97g, 18.12) was added mmol, 2.3 mL), after stirring for 30 min, benzoyl chloride (968.00 mg, 6.89 mmol, 0.8 mL) was added, and the reaction was stirred at 20 ° C. for 3 h. The reaction solution was quenched with water (10 mL) and ammonia water (10 mL), stirred for 30 min and extracted with ethyl acetate (20 mL x 3).
- reaction solution was diluted with dichloromethane (20 mL), filtered, and the filtrate was washed with water (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude product 5-19, which was directly used in the next reaction without further purification.
- compound D-arabinose 25g, 166.5mmol
- DMF 250mL
- imidazole 17g, 249.8mmol
- tert-butyldiphenylchlorosilane 45.8g, 166.5mmol
- the solution was poured into water (2.5 L), extracted with ethyl acetate (1000 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude product 6-1, which was directly used in the next reaction without further purification.
- compound 6-4 (1.24g, 3.6mmol) was dissolved in dichloromethane (40mL), followed by pyridine (2.7mL 33.5mmol) and trifluoromethanesulfonic anhydride (0.8mL, 4.9mmol), 20min After that, water (10 mL) was added to quench the reaction, and the liquid was separated. The organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in tetrahydrofuran (25 mL) and cooled to 0 ° C.
- reaction solution was diluted with ethyl acetate (50 mL), filtered, and the filtrate was washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure.
- Fluorescence polarization test (fluorescence polarization assay, FP assay) was used to detect the affinity of compounds for human STING protein. There is a certain amount of fluorescein-labeled c-di-GMP and different concentrations of test compounds in the reaction system. When the recombinant human STING C-terminal protein is added, the two small molecules competitively bind to the protein. The bound fluorescein-labeled c-di-GMP rotates slowly in the liquid phase, and the degree of fluorescence polarization detected at this time is also high. The degree of fluorescence polarization is inversely proportional to the concentration and affinity of the test compound. By detecting the magnitude of polarized light in the reaction system, we can accurately know the affinity of the test compound for human STING.
- the soluble human STING protein sequence used in the experiment was intercepted from the C-terminal part of human wild-type endoplasmic reticulum binding protein STING, from 140 amino acids to 379 amino acids.
- the human STING protein has a variety of alleles with different sequence differences. Different alleles have different affinity for CDN (Yi, et.al., "Single Nucleotide Polymorphisms of Human STING can affect the immune response" to cyclic dinucleotides "PLOSONE.2013, 8 (10), e77846). Wild-type STING sequences (G230, R232, R293) account for approximately 57.9% of the total.
- the N-terminus of the recombinant STING protein is a 6His-SUMO sequence, which facilitates the correct folding and purification of the protein, which is excised by proteases, and the C-terminus STING is used for FP testing.
- FP test a 384-well plate was used, and 10 ⁇ l of reaction system was added with a final concentration of 30 nM fluorescein-labeled c-di-GMP, 10 ⁇ M human STING protein, and different concentrations of reference compound or test compound. Centrifuge at 1000g for 1 minute, incubate at room temperature in the dark for 30 minutes, and read the plate with Envision.
- the THP1-Dual TM cells (InvivoGen catalog code: thpd-nfis) used in the test were constructed by stably integrating two inducible reporter genes in the human monocyte cell line THP1.
- the promoter sequence of the secreted embryonic alkaline phosphatase (SEAP) reporter gene consists of an IFN- ⁇ basic promoter and 5 copies of the upstream NF- ⁇ B consensus transcriptional response element (NF- ⁇ B consensus transcriptional response element) And 3 copies of the c-Rel binding site.
- SEAP secreted embryonic alkaline phosphatase
- the secreted luciferase (Lucia) reporter gene is driven by five interferon (IFN) -stimulated response elements and an ISG54 basic promoter. This makes it possible to study the two main downstream signaling pathways of STING at the same time: to study the NF ⁇ B pathway by detecting SEAP activity; and to study the IRF pathway by evaluating the activity of Lucia luciferase
- the compound was diluted with PB buffer (50 mM HEPES, 100 mM KCl, 3 mM MgCl2, 0.1 mM DTT, 85 mM Sucrose, 1 mM ATP, 0.1 mM GTP, 0.2% BSA).
- PB buffer 50 mM HEPES, 100 mM KCl, 3 mM MgCl2, 0.1 mM DTT, 85 mM Sucrose, 1 mM ATP, 0.1 mM GTP, 0.2% BSA.
- PB buffer 50 mM HEPES, 100 mM KCl, 3 mM MgCl2, 0.1 mM DTT, 85 mM Sucrose, 1 mM ATP, 0.1 mM GTP, 0.2% BSA.
- the compound of the present invention has a strong ability to promote ⁇ -interferon activation.
- the RAW-Dual TM cells (InvivoGen catalog code: rawd-ismip) used in the test were constructed by stably integrating two inducible reporter genes in the mouse macrophage cell line RAW264.7: the NF-KB pathway was studied by detecting SEAP activity, And study the IRF3 pathway by evaluating Lucia luciferase activity.
- Cell suspension (50,000 cells per well) was added to a 96-well plate (Corning 3599 flat bottom plate) at 200 ⁇ L per well, and cultured in a 37 ° C incubator for 18 to 24 hours.
- 4T1 breast cancer homologous mouse model was used to evaluate the efficacy of the compound.
- 1E5 4T1 breast cancer cells (Shanghai Institutes of Chinese Academy of Sciences) were inoculated subcutaneously in 6-8 week old Balb / C mice (Viton Lihua). When the tumor volume reached 100 mm3, they were randomly divided into 8 groups. On the 1st, 4th, and 8th days after grouping, intratumoral administration was performed sequentially. A single intratumoral administration (IT) is the first day after grouping.
- the ADU-S100 dose group was 100ug per mouse (single) and 30ug per mouse (three times).
- CT-26 colon cancer homologous mouse model was used to evaluate the efficacy of the compound.
- 3E5 CT-26 colon cancer cells (ATCC-CRL-2638) were inoculated subcutaneously in 6-8 week old Balb / C mice (Shanghai Lingchang Biology), when the tumor volume reached 100mm3, they were randomly divided into groups of 8 .
- intratumoral administration was performed three times in total.
- the dosage of compound 2B in each group was 1ug per mouse, 3ug per mouse, 9ug per mouse, and 18 ⁇ g per mouse.
- Compound ADU-S100 was administered at a dose of 125 ⁇ g per mouse.
- Tumor volume was measured three times a week after the start of dosing.
- SEM standard error
- the MC38 colon cancer mouse model was used to evaluate the efficacy of the compound.
- 3E5 MC38 colon cancer cells (Nanjing Kebai) were inoculated subcutaneously in 6-8 week-old C57BL / 6 mice (Shanghai Bikai). When the tumor volume reached about 100 mm3, they were randomly divided into groups of 5-6 mice. On the 1st, 4th, and 8th days after grouping, intratumoral administration was performed sequentially. A single intratumoral administration is the first day after grouping.
- the ADU-S100 dose group is 100ug per mouse (single).
- the compound 2B dose group was 100 ug per mouse (single), 30 ug per mouse (three times) and 10 ug per mouse (three times).
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Abstract
提供了式(Ⅰ)所示化合物、其光学异构体及其药效上可接受的盐，以及该化合物作为STING激动剂的应用。
Description
本申请主张如下优先权：
CN201811188184.3，申请日2018年10月12日；
CN201811301854.8，申请日2018年11月2日；
CN201811367721.0，申请日2018年11月16日；
CN201910129734.2，申请日2019年2月21日；
CN201910463705.X，申请日2019年5月30日。
本发明涉及式(Ⅰ)所示化合物、其光学异构体及其药效上可接受的盐，以及该化合物作为STING激动剂的应用。
长久以来，科研人员一直试图通过激活病人的免疫系统，使他们自身的免疫系统能够有效地对抗肿瘤，完全清除肿瘤细胞。但肿瘤自发缓解的概率极低，因此绝大多数病人也无法因此获益。上世纪六七十年代，出现了通过卡介苗注射，非特异性的强化免疫系统功能等治疗方法。八十年代，能够活化T细胞以及NK细胞的干扰素和IL-2也被尝试应用于癌症的治疗，但这些方法依然有非常多的局限性，诸如外源细胞因子在血液中的半寿期非常短，这必须采用频繁给药和高剂量予以补偿。非特异性活化免疫系统导致正常组织的炎症反应，细胞因子风暴等，因此很多疗法的毒副作用非常强。作为在体中触发具有特异性治疗有益性细胞因子产生的免疫调节剂，以STING为靶标的疗法为解决这一困境带来了曙光。
目前己知人STING以三种方式激活：1)通过结合正在侵入的细菌或古细菌释放的外源(3’，3’)环状二核苷酸(c-diGMP、c-diAMP和c-GAMP)激活，这显示了STING具有在抗感染中先天免疫活化的作用；2)通过结合(2’3’)环状鸟苷单磷酸腺苷单磷酸酯(2’,3’c-GAMP)激活，它是由环状GMP-AMP二核苷酸合成酶(cGAS)在外源双链DNA(例如由正在侵入的细菌、病毒或原虫释放的)或哺乳动物中的自我DNA存在时诱导产生的内源环状二核苷酸，这显示了STING具有受内源或外源DNA诱导活化先天免疫的作用；3)通过结合合成性配体活化。
STING作为细胞质中DNA的感受器，它的活化可导致下游IRF3和NF-κB两条通路的激活以激活免疫系统。NF-κB通路激活导致下游一系列致炎症细胞因子的活化，而IRF3通路的激活，导致了一型干扰素(IFN-α/β)的激活,树突状细胞，细胞毒性细胞，NK细胞等的活化，从而发挥出抗肿瘤作用。
人体内的DNA通常不会激活STING蛋白，因为正常情况下DNA仅能够存在于细胞核之内(线粒体DNA除外)。但如果DNA泄漏到胞浆之中，则会活化STING，引发免疫反应。最近发现放疗以及化疗同样能够激活STING，这可能也是由于死亡的肿瘤细胞内的DNA泄漏导致STING被激活。
发明内容
本发明提供了式(Ⅰ)所示化合物、其光学异构体及其药效上可接受的盐，
其中，
R
1、R
1a分别独立地选自
T
1、T
2、T
3、T
4、T
5、T
6、T
7、T
8、T
9、T
10、T
11、T
12、T
13分别独立地选自-C(R)-和-N-；
L
1、L
2分别独立地选自-O-、-N(R)-、-C(RR)-和-C(＝O)-；
R分别独立地选自H、卤素、OH、NH
2、CN、
C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R’取代；
R’选自F、Cl、Br、I、OH、NH
2和CH
3；
R
2、R
2a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
3、R
3a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
4、R
4a分别独立地选自H、卤素、OH、NH
2、CN、N
3、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
5、R
5a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
6、R
6a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R取代；
R
7、R
7a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R取代；
R
10、R
10a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨 基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R取代；
或者，R
7和R
10连接在一起，形成一个C
3-6环烷基、C
3-6环稀基或C
3-6环炔基，所述C
3-6环烷基、C
3-
6环稀基或C
3-6环炔基任选被1、2或3个R取代；
R
7a和R
10a连接在一起，形成一个C
3-6环烷基、C
3-6环稀基或C
3-6环炔基，所述C
3-6环烷基、C
3-6环稀基或C
3-6环炔基任选被1、2或3个R取代；
R
8选自BH
3
-和-S(R
9)；
R
9选自H、CH
2OC(＝O)R
11、CH
2OC(＝O)OR
11、CH
2CH
2SC(＝O)R
11和CH
2CH
2SSCH
2R
11；
R
11选自C
6-10芳基、5～10元杂芳基、C
1-6杂环烷基和C
1-
20烷基，所述C
1-
20烷基任选被1、2、3、4或5个C
6-10芳基、C
3-10环烷基、OH和F取代；
或者，R
4与R
6或R
4a与R
6a连接在一起形成一个5～6元杂环烷基；
X
1、X
1a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
2、X
2a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
3、X
3a分别独立地选自-O-和-S-；
Y、Y
a分别独立地选自-O-、-S-、-CH
2-和-C(＝CH
2)-；
所述5～6元杂环烷基、5～10元杂芳基或C
1-6杂环烷基包含1、2或3个独立选自-O-、-NH-、-S-、-C(＝O)-、-C(＝O)O-、-S(＝O)-、-S(＝O)
2-和N的杂原子或杂原子团；
且，当R
1或R
1a选自
时，式(I)所示的化合物不选自
本发明的一些方案中，上述当R
8选自BH
3
-时，R
4和R
4a其中一个选自F、Cl和Br，另一个选自F、Cl、Br、OH、OCH
3或N
3，其他变量如本发明所定义。
本发明的一些方案中，上述R分别独立地选自H、卤素、OH、NH
2、CN、
C
1-3烷基、C
1-3烷氧基、C
1-3烷硫基和C
1-3烷氨基，其中C
1-3烷基、C
1-3烷氧基、C
1-3烷硫基和C
1-3烷氨基任选被1、2或3个R’取代，其他变量如本发明所定义。
本发明的一些方案中，上述R分别独立地选自H、F、Cl、Br、I、OH、NH
2、CN、Me、
其中Me、
任选被1、2或3个R’取代，其他变量如本发明所定义。
本发明的一些方案中，上述R分别独立地选自H、F、Cl、Br、I、OH、NH
2、CN、Me、
其他变量如本发明所定义。
本发明的一些方案中，上述R
1、R
1a分别独立地选自
其他变量如本发明所定义。
本发明的一些方案中，上述R
1、R
1a分别独立地选自
其他变量如本发明所定义。
本发明的一些方案中，上述R
2、R
2a、R
3、R
3a、R
5和R
5a、R
6和R
6a分别独立地选自H，其他变量如本发明所定义。
本发明的一些方案中，上述R
6和R
6a分别独立地选自H和甲基，其他变量如本发明所定义。
本发明的一些方案中，上述R
4、R
4a分别独立地选自F、OH、NH
2、N
3和
其他变量如本发明所定义。
本发明的一些方案中，上述R
7、R
7a分别独立地选自H和CH
3，其他变量如本发明所定义。
本发明的一些方案中，上述R
4与R
6连接在一起，结构单元
选自
其他变量如本发明所定义。
本发明的一些方案中，上述R
4a与R
6a连接在一起，结构单元
选自
其他变量如本发明所定义。
本发明的一些方案中，上述化合物、其光学异构体及其药学上可接受的盐，其选自
其中，
R
1、R
1a分别独立地选自
R
4、R
4a分别独立地选自H、卤素、OH、NH
2、CN、N
3、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
6选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R取代；
或者，R
4与R
6连接在一起形成一个5～6元杂环烷基；
R
8选自BH
3
-和-S(R
9)；
R
9选自H、CH
2OC(＝O)R
11、CH
2OC(＝O)OR
11、CH
2CH
2SC(＝O)R
11和CH
2CH
2SSCH
2R
11；
R
11选自C
6-10芳基、5～10元杂芳基、C
1-6杂环烷基和C
1-
20烷基，所述C
1-
20烷基任选被1、2、3、4或5个C
6-10芳基、C
3-10环烷基、OH和F取代；
X
1、X
1a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
2、X
2a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
3、X
3a分别独立地选自-O-和-S-；
Y、Y
a分别独立地选自-O-、-S-、-CH
2-和-C(＝CH
2)-；
当R
8选自BH
3
-时，R
4和R
4a其中一个选自F、Cl和Br，另一个选自F、Cl、Br、OH、OCH
3或N
3；
当R
8选自-S(R
9)时，R
4和R
4a其中一个选自F、Cl和Br，另一个选自OH、OCH
3或N
3；
或者，当R
8选自-S(R
9)，且R
4和R
4a其中一个选自F、Cl和Br，另一个不选自OH、OCH
3或N
3时，R
4和R
4a不选自
且R
4和R
4a不同时选自
本发明的一些方案中，上述化合物、其光学异构体及其药效上可接受的盐，其选自
其中，R
1、R
1a、R
4a、R
7、R
7a、R
8、R
6a如上述所定义。
本发明的一些方案中，上述化合物、其光学异构体及其药效上可接受的盐，其选自
本发明的一些方案中，上述化合物、其光学异构体及其药学上可接受的盐，其选自
其中，
R
1、R
1a分别独立地选自
X
1、X
1a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
2、X
2a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
3、X
3a分别独立地选自-O-和-S-；
Y、Y
a分别独立地选自-O-、-S-、-CH
2-和-C(＝CH
2)-。
本发明的一些方案中，上述化合物、其光学异构体及其药学上可接受的盐，其选自
本发明的一些方案中，上述R
1、R
1a分别独立地选自
其他变量如本发明所定义。
本发明提供了式(Ⅰ)所示化合物、其光学异构体及其药效上可接受的盐，
其中，
R
1、R
1a分别独立地选自
T
1、T
2、T
3、T
4、T
5、T
6、T
7、T
8、T
9、T
10、T
11、T
12、T
13分别独立地选自-C(R)-和-N-；
L
1、L
2分别独立地选自-O-、-N(R)-、-C(RR)-和-C(＝O)-；
R分别独立地选自H、卤素、OH、NH
2、CN、
C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R’取代；
R’选自F、Cl、Br、I、OH、NH
2和CH
3；
R
2、R
2a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-
6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
3、R
3a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-
6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
4、R
4a分别独立地选自H、卤素、OH、NH
2、CN、N
3、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
5、R
5a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-
6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-6炔基任选被1、2或3个R取代；
R
6、R
6a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基、C
1-6烷氨基和C
2-
6炔基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R取代；
R
7、R
7a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R取代；
R
10、R
10a分别独立地选自H、卤素、OH、NH
2、CN、C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基，其中C
1-6烷基、C
1-6烷氧基、C
1-6烷硫基和C
1-6烷氨基任选被1、2或3个R取代；
或者，R
7和R
10连接在一起，形成一个C
3-6环烷基、C
3-6环稀基或C
3-6环炔基，所述C
3-6环烷基、C
3-6环稀基或C
3-6环炔基任选被1、2或3个R取代；
R
7a和R
10a连接在一起，形成一个C
3-6环烷基、C
3-6环稀基或C
3-6环炔基，所述C
3-6环烷基、C
3-6环稀基或C
3-6环炔基任选被1、2或3个R取代；
R
8选自BH
3
-和-S(R
9)；
R
9选自H、CH
2OC(＝O)R
11、CH
2OC(＝O)OR
11、CH
2CH
2SC(＝O)R
11和CH
2CH
2SSCH
2R
11；
R
11选自C
6-10芳基、5～10元杂芳基、C
1-6杂环烷基和C
1-
20烷基，所述C
1-
20烷基任选被1、2、3、4或5个C
6-10芳基、C
3-10环烷基、OH和F取代；
或者，R
4与R
6或R
4a与R
6a连接在一起形成一个5～6元杂环烷基；
X
1、X
1a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
2、X
2a分别独立地选自-NH-、-O-、-S-和-CH
2-；
X
3、X
3a分别独立地选自-O-和-S-；
Y、Y
a分别独立地选自-O-、-S-、-CH
2-和-C(＝CH
2)-；
所述5～6元杂环烷基、5～10元杂芳基或C
1-6杂环烷基包含1、2或3个独立选自-O-、-NH-、-S-、-C(＝O)-、-C(＝O)O-、-S(＝O)-、-S(＝O)
2-和N的杂原子或杂原子团。
本发明的一些方案中，上述R分别独立地选自H、卤素、OH、NH
2、CN、
C
1-3烷基、C
1-3烷氧基、C
1-3烷硫基和C
1-3烷氨基，其中C
1-3烷基、C
1-3烷氧基、C
1-3烷硫基和C
1-3烷氨基任选被1、2或3个R’取代。
本发明的一些方案中，上述R分别独立地选自H、F、Cl、Br、I、OH、NH
2、CN、Me、
其中Me、
任选被1、2或3个R’取代。
本发明的一些方案中，上述R分别独立地选自H、F、Cl、Br、I、OH、NH
2、CN、Me、
本发明的一些方案中，上述R
1、R
1a分别独立地选自
本发明的一些方案中，上述R
1、R
1a分别独立地选自
本发明的一些方案中，上述R
2、R
2a、R
3、R
3a、R
5、R
5a、R
6和R
6a分别独立地选自H。
本发明的一些方案中，上述R
4、R
4a分别独立地选自F、OH、NH
2、N
3和
本发明的一些方案中，上述R
7、R
7a分别独立地选自H和CH
3。
本发明的一些方案中，上述R
4与R
6连接在一起，结构单元
选自
本发明的一些方案中，上述R
4a与R
6a连接在一起，结构单元
选自
本发明的一些方案中，上述化合物、其光学异构体及其药效上可接受的盐，其选自
其中，
R
1、R
1a、R
4a、R
7、R
7a、R
8和R
6a如上述所定义。
本发明还提供了下式化合物、其光学异构体及其药效上可接受的盐，其选自
定义和说明
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物的中性形式接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机胺或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物的中性形式接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，所述无机酸包括例如盐酸、氢溴酸、硝酸、碳酸，碳酸氢根，磷酸、磷酸一氢根、磷酸二氢根、硫酸、硫酸氢根、氢碘酸、亚磷酸等；以及有机酸盐，所述有机酸包括如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸和甲磺酸等类似的酸；还包括氨基酸(如精氨酸等)的盐，以及如葡糖醛酸等有机酸的盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。
除非另有说明，用楔形实线键
和楔形虚线键
表示一个立体中心的绝对构型，用直形实线键
和直形虚线键
表示立体中心的相对构型，用波浪线
表示楔形实线键
或楔形虚线键
或用波浪线
表示直形实线键
和直形虚线键
本发明的化合物可以存在特定的。除非另有说明，术语“互变异构体”或“互变异构体形式”是指在室温下，不同官能团异构体处于动态平衡，并能很快的相互转化。若互变异构体是可能的(如在溶液中)，则可以达到互变异构体的化学平衡。例如，质子互变异构体(proton tautomer)(也称质子转移互变异构体(prototropic tautomer))包括通过质子迁移来进行的互相转化，如酮-烯醇异构化和亚胺-烯胺异构化。价键异构体(valence tautomer)包括一些成键电子的重组来进行的相互转化。其中酮-烯醇互变异构化的具体实例是戊烷-2,4-二酮与4-羟基戊-3-烯-2-酮两个互变异构体之间的互变，或例如
为互变异构体。
除非另有说明，术语“富含一种异构体”、“异构体富集”、“富含一种对映体”或者“对映体富集”指其中一种异构体或对映体的含量小于100％，并且，该异构体或对映体的含量大于等于60％，或者大于等于70％，或者大于等于80％，或者大于等于90％，或者大于等于95％，或者大于等于96％，或者大于等于97％，或者大于等于98％，或者大于等于99％，或者大于等于99.5％，或者大于等于99.6％，或者大于等于99.7％，或者大于等于99.8％，或者大于等于99.9％。
除非另有说明，术语“异构体过量”或“对映体过量”指两种异构体或两种对映体相对百分数之间的差值。例如，其中一种异构体或对映体的含量为90％，另一种异构体或对映体的含量为10％，则异构体或对映体过量(ee值)为80％。
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚(
3H)，碘-125(
125I)或C-14(
14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药 物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。
术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，可以包括重氢和氢的变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧(即＝O)时，意味着两个氢原子被取代。氧取代不会发生在芳香基上。术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。
当所列举的连接基团没有指明其连接方向，其连接方向是任意的，例如，
中连接基团L为-M-W-，此时-M-W-既可以按与从左往右的读取顺序相同的方向连接苯环和环戊烷构成
也可以按照与从左往右的读取顺序相反的方向连接苯环和环戊烷构成
所述连接基团、取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。
除非另有规定，环上原子的数目通常被定义为环的元数，例如，“5-6元环”是指环绕排列5-6个原子的“环”。
除非另有规定，“5-6元环”表示由5至6个环原子组成的环烷基、杂环烷基、环烯基、杂环烯基、环炔基、杂环炔基、芳基或杂芳基。所述的环包括单环，也包括螺环、并环和桥环等双环体系。除非另有规定，该环任选地包含1、2或3个独立选自O、S和N的杂原子。所述5-6元环包括5元、6元环等。“5-6元环”包括例如苯基、吡啶基和哌啶基等；另一方面，术语“5-6元杂环烷基”包括哌啶基等，但不包括苯基。术语“环”还包括含有至少一个环的环系，其中的每一个“环”均独立地符合上述定义。
除非另有规定，术语“C
1-20烷基”用于表示直链或支链的由1至20个碳原子组成的饱和碳氢基团。所述C
1-20烷基包括C
1-10、C
1-9、C
1-8、C
1-6、C
1-5、C
1-14、C
1-3、C
1-2、C
2-16、C
2-4、C
10、C
8、C
7、C
6和C
5烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C
1-20烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)、戊基(包括n-戊基，异戊基和新戊基)、己基、庚基、辛基等。
除非另有规定，术语“C
1-6烷基”用于表示直链或支链的由1至6个碳原子组成的饱和碳氢基团。所述C
1-6烷基包括C
1-5、C
1-4、C
1-3、C
1-2、C
2-6、C
2-4、C
6和C
5烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C
1-6烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)、戊基(包括n-戊基，异戊基和新戊基)、己基等。
除非另有规定，术语“C
1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C
1-3烷基包括C
1-2和C
2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C
1-
3烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)等。
术语“杂烷基”本身或者与另一术语联合，表示由一定数目碳原子和至少一个杂原子或杂原子团组成的，稳定的直链或支链的烷基原子团或其组合物。在一些实施方案中，杂原子选自B、O、N和S，其中氮和硫原子任选地被氧化，氮杂原子任选地被季铵化。在另一些实施方案中，杂原子团选自-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)、-S(＝O)
2-、-C(＝O)N(H)-、-N(H)-、-C(＝NH)-、-S(＝O)
2N(H)-和-S(＝O)N(H)-。在一些实施方案中，所述杂烷基为C
1-6杂烷基；在另一些实施方案中，所述杂烷基为C
1-3杂烷基。杂原子或杂原子团可以位于杂烷基的任何内部位置，包括该烷基与分子其余部分的连接位置，但术语“烷氧基”、“烷氨基”和“烷硫基”(或硫代烷氧基)属于惯用表达，是指分别通过一个氧原子、氨基或硫原子连接到分子的其余部分的那些烷基基团。杂烷基的实例包括但不限于-OCH
3、-OCH
2CH
3、-OCH
2CH
2CH
3、-OCH
2(CH
3)
2、-CH
2-CH
2-O-CH
3、-NHCH
3、-N(CH
3)
2、-NHCH
2CH
3、-N(CH
3)(CH
2CH
3)、-CH
2-CH
2-NH-CH
3、-CH
2-CH
2-N(CH
3)-CH
3、-SCH
3、-SCH
2CH
3、-SCH
2CH
2CH
3、-SCH
2(CH
3)
2、-CH
2-S-CH
2-CH
3、-CH
2-CH
2、-S(＝O)-CH
3、-CH
2-CH
2-S(＝O)
2-CH
3、和。至多两个杂原子可以是连续的，例如-CH
2-NH-OCH
3。
除非另有规定，术语“C
1-6烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至6个碳原子的烷基基团。所述C
1-6烷氧基包括C
1-4、C
1-3、C
1-2、C
2-6、C
2-4、C
6、C
5、C
4和C
3烷氧基等。C
1-6烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)、丁氧基(包括n-丁氧基、异丁氧基、s-丁氧基和t-丁氧基)、戊氧基(包括n-戊氧基、异戊氧基和新戊氧基)、己氧基等。
除非另有规定，术语“C
1-3烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至3个碳原子的烷基基团。所述C
1-3烷氧基包括C
1-2、C
2-3、C
3和C
2烷氧基等。C
1-3烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)等。
除非另有规定，术语“C
1-6烷氨基”表示通过氨基连接到分子的其余部分的那些包含1至6个碳原子的烷基基团。所述C
1-6烷氨基包括C
1-4、C
1-3、C
1-2、C
2-6、C
2-4、C
6、C
5、C
4、C
3和C
2烷氨基等。C
1-6烷氨基的实例包括但不限于-NHCH
3、-N(CH
3)
2、-NHCH
2CH
3、-N(CH
3)CH
2CH
3、-N(CH
2CH
3)(CH
2CH
3)、-NHCH
2CH
2CH
3、-NHCH
2(CH
3)
2、-NHCH
2CH
2CH
2CH
3等。
除非另有规定，术语“C
1-3烷氨基”表示通过氨基连接到分子的其余部分的那些包含1至3个碳原子的烷基基团。所述C
1-3烷氨基包括C
1-2、C
3和C
2烷氨基等。C
1-3烷氨基的实例包括但不限于-NHCH
3、-N(CH
3)
2、-NHCH
2CH
3、-N(CH
3)CH
2CH
3、-NHCH
2CH
2CH
3、-NHCH
2(CH
3)
2等。
除非另有规定，术语“C
1-6烷硫基”表示通过硫原子连接到分子的其余部分的那些包含1至6个碳原子的烷基基团。所述C
1-6烷硫基包括C
1-4、C
1-3、C
1-2、C
2-6、C
2-4、C
6、C
5、C
4、C
3和C
2烷硫基等。C
1-6烷硫基的实例包括但不限于-SCH
3、-SCH
2CH
3、-SCH
2CH
2CH
3、-SCH
2(CH
3)
2等等。
除非另有规定，术语“C
1-3烷硫基”表示通过硫原子连接到分子的其余部分的那些包含1至3个碳原子的烷基基团。所述C
1-3烷硫基包括C
1-3、C
1-2和C
3烷硫基等。C
1-3烷硫基的实例包括但不限于-SCH
3、-SCH
2CH
3、-SCH
2CH
2CH
3、-SCH
2(CH
3)
2等。
除非另有规定，“C
2-6炔基”用于表示直链或支链的包含至少一个碳-碳三键的由2至6个碳原子组成的碳氢基团，碳-碳三键可以位于该基团的任何位置上。所述C
2-6炔基包括C
2-4、C
2-3、C
4、C
3和C
2炔基等。其可以是一价、二价或者多价。C
2-6炔基的实例包括但不限于乙炔基、丙炔基、丁炔基、戊炔基等。
除非另有规定，术语“5-6元杂环烷基”本身或者与其他术语联合分别表示由5至6个环原子组成的饱和环状基团，其1、2、3或4个环原子为独立选自O、S和N的杂原子，其余为碳原子，其中氮原子任选地被季铵化，氮和硫杂原子可任选被氧化(即NO和S(O)
p，p是1或2)。其包括单环和双环体系，其中双环体系包括螺环、并环和桥环。此外，就该“5-6元杂环烷基”而言，杂原子可以占据杂环烷基与分子其余部分的连接位置。所述5-6元杂环烷基包括5元和6元杂环烷基。5-6元杂环烷基的实例包括但不限于吡咯烷基、吡唑烷基、咪唑烷基、四氢噻吩基(包括四氢噻吩-2-基和四氢噻吩-3-基等)、四氢呋喃基(包括四氢呋喃-2-基等)、四氢吡喃基、哌啶基(包括1-哌啶基、2-哌啶基和3-哌啶基等)、哌嗪基(包括1-哌嗪基和2-哌嗪基等)、吗啉基(包括3-吗啉基和4-吗啉基等)、二噁烷基、二噻烷基、异噁唑烷基、异噻唑烷基、1,2-噁嗪基、1,2-噻嗪基、六氢哒嗪基、高哌嗪基或高哌啶基等。
除非另有规定，本发明术语“C
6-10芳环”和“C
6-10芳基”可以互换使用，术语“C
6-10芳环”或“C
6-10芳基”表示由6至10个碳原子组成的具有共轭π电子体系的环状碳氢基团，它可以是单环、稠合双环或稠合三环体系，其中各个环均为芳香性的。其可以是一价、二价或者多价，C
6-10芳基包括C
6-9、C
9、C
10和C
6芳基等。C
6-10芳基的实例包括但不限于苯基、萘基(包括1-萘基和2-萘基等)。
除非另有规定，本发明术语“5-10元杂芳环”和“5-10元杂芳基”可以互换使用，术语“5-10元杂芳基”是表示由5至10个环原子组成的具有共轭π电子体系的环状基团，其1、2、3或4个环原子为独立选自O、S和N的杂原子，其余为碳原子。其可以是单环、稠合双环或稠合三环体系，其中各个环均为芳香性的。其中氮原子任选地被季铵化，氮和硫杂原子可任选被氧化(即NO和S(O)
p，p是1或2)。5-10元杂芳基可通过杂原子或碳原子连接到分子的其余部分。所述5-10元杂芳基包括5-8元、5-7元、5-6元、5元和6元杂芳基等。所述5-10元杂芳基的实例包括但不限于吡咯基(包括N-吡咯基、2-吡咯基和3-吡咯基等)、吡唑基(包括2-吡唑基和3-吡唑基等)、咪唑基(包括N-咪唑基、2-咪唑基、4-咪唑基和5-咪唑基等)、噁唑基(包括2-噁唑基、4-噁唑基和5-噁唑基等)、三唑基(1H-1,2,3-三唑基、2H-1,2,3-三唑基、1H-1,2,4-三唑基和4H-1,2,4-三唑基等)、四唑基、异噁唑基(3-异噁唑基、4-异噁唑基和5-异噁唑基等)、噻唑基(包括2-噻唑基、4-噻唑基和5-噻唑基等)、呋喃基(包括2-呋喃基和3-呋喃基等)、噻吩基(包括2-噻吩基和3-噻吩基等)、吡啶基(包括2-吡啶基、3-吡啶基和4-吡啶基等)、吡嗪基、嘧啶基(包括2-嘧啶基和4-嘧啶基等)、苯并噻唑基(包括5-苯并噻唑基等)、嘌呤基、苯并咪唑基(包括2-苯并咪唑基等)、苯并噁唑基、吲哚基(包括5-吲哚基等)、异喹啉基(包括1-异喹啉基和5-异喹啉基等)、喹喔啉基(包括2-喹喔啉基和5-喹喔啉基等)或喹啉基(包括3-喹啉基和6-喹啉基等)。
除非另有规定，C
n-n+m或C
n-C
n+m包括n至n+m个碳的任何一种具体情况，例如C
1-6包括C
1、C
2、C
3、C
4、C
5和C
6，也包括n至n+m中的任何一个范围，例如C
1-6包括C
1-3、C
1-6、C
1-4、C
3-6、C
3-5、C
2-5和C
1-
5等；同理，n元至n+m元表示环上原子数为n至n+m个，例如5-6元环包括5元环和6元环。
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。
本发明HPLC检测条件主要如下:色谱柱:YMC-Pack ODS-A 150*4.6mm,5μm,流动相:水(0.06875％三氟乙酸)-乙腈(0.0625％三氟乙酸)；流速：1.0mL/min；检测波长：UV 220nm&215nm&254nm；柱 温:40℃。
本发明采用下述缩略词：aq代表水；CDCl
3代表氘代氯仿；CD
3OD代表氘代甲醇；DMSO-d
6代表氘代二甲亚砜；DMF代表N,N-二甲基甲酰胺；Bz代表苯甲酰基；TBS代表叔丁基二甲基硅基；DMTr代表4,4'-双甲氧基三苯甲基；CE代表氰乙基；i-Pr代表异丙基；DMTrCl代表4,4'-双甲氧基三苯甲基氯；DDTT代表(E)-N,N-二甲基-N'-(3-硫代-3H-1,2,4-二硫唑-5-基)甲脒；DCA代表2,2-二氯乙酸；BSA代表N,O-双三甲硅基乙酰胺，ug或μg代表微克。
图1：4T1乳腺癌同系小鼠模型药效实验结果；
图2：CT-26结肠癌同系小鼠模型药效实验结果；
图3：MC38结肠癌同系小鼠模型药效实验结果。
下面通过实施例对本申请进行详细描述，但并不意味着存在对本申请而言任何不利的限制。本文已经详细地描述了本申请，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本申请精神和范围的情况下针对本申请具体实施方式进行各种变化和改进将是显而易见的。
实施例1：化合物1A、1B、1C和1D的制备
步骤1：化合物1-2的制备
氮气保护下，向化合物1-1(1g,3.71mmol)的吡啶(20mL)溶液中滴加三甲基氯硅烷(1.61g,14.86mmol,1.89mL)，0℃反应30min后，将苯甲酰氯(605mg,4.30mmol,500.00μL)加入反应体系，升温至15℃反应3h。停止反应，降温至0℃，依次加入水(10mL)、氨水(5mL)淬灭反应，搅拌10min，反应混合物用乙酸乙酯(25mL x 3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝9/1)，得化合物1-2。
MS(ESI)m/z(M+H)
+＝374.1.
1H NMR(400MHz,DMSO-d
6)δ11.26(br s,1H),8.77(s,1H),8.71(s,1H),8.05(d,J＝7.3Hz,2H),7.66(t,J＝7.6Hz,2H),7.56(t,J＝7.6Hz,2H),6.39(dd,J＝2.3,17.3Hz,1H),5.77(d,J＝6.3Hz,1H),5.64-5.39(m,1H),5.18(t,J＝5.4Hz,1H),4.63-4.45(m,1H),4.08-3.93(m,1H),3.81-3.76(m,1H),3.64-3.58(m,1H).
步骤2：化合物1-3的制备
将化合物1-2(1.5g,4.02mmol)溶于吡啶(15mL)中，依次加入硝酸银(2.73g,16.07mmol)、叔丁基二甲基氯硅烷(636mg,4.22mmol,517.07μL)，反应体系室温搅拌4h。反应加水(50mL)淬灭，乙酸乙酯(40mL x 3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝4/1)，得化合物1-3。
MS(ESI)m/z(M+H)
+＝488.1.
1H NMR(400MHz,DMSO-d
6)δ11.29(br s,1H),8.73(br s,1H),8.58(s,1H),8.01(br d,J＝7.3Hz,2H),7.66-7.36(m,3H),6.39(br d,J＝18.6Hz,1H),5.82(d,J＝6.6Hz,1H),5.66-5.39(m,1H),4.73-4.50(m,1H),4.08-4.02(m,1H),4.00-3.94(m,1H),3.85-3.80(m,1H),0.83(s,9H),0.03(s,3H),0.00(s,3H).
步骤3：化合物1-4的制备
将化合物1-3(1.8g,3.69mmol)溶于二氯甲烷(20mL)中，依次加入2,4,6-三甲基吡啶(3.30g,27.24mmol,3.60mL)、4,4'-双甲氧基三苯甲基氯(3.75g,11.07mmol)，反应体系室温搅拌16h。反应加水(20mL)淬灭，乙酸乙酯(20mL x 3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝7/3)，得化合物1-4。
MS(ESI)m/z(M+H)
+＝790.4.
1H NMR(400MHz,CDCl
3)δ9.04(s,1H),8.82(s,1H),8.19(s,1H),8.06(d,J＝7.2Hz,1H),7.72-7.62(m,1H),7.59-7.54(m,4H),7.49-7.43(m,4H),7.35-7.31(m,2H),7.29-7.24(m,1H),6.86-6.83(m,4H),6.37(dd,J＝2.9,15.2Hz,1H),4.66-4.57(m,1H),4.54-4.40(m,1H),4.14(br s,1H),3.81(s,3H),3.80(s,3H),3.79-3.70(m,1H),3.49(dd,J＝3.2,11.7Hz,1H),0.83(s,9H),0.01(s,3H),0.00(s,3H).
步骤4：化合物1-5的制备
0℃条件下，将氟化四丁基铵的四氢呋喃溶液(1M,3.06mL)加入化合物1-4(2.2g,2.78mmol)的四氢呋喃(20mL)溶液中，加毕，反应升温至室温搅拌反应3h。反应体系倾入乙酸乙酯(100mL)和水(100mL)组成的混合体系中，有机相依次用水(50mL x 3)，饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝9/1)，得化合物1-5。
MS(ESI)m/z(M+H)
+＝676.2.
1H NMR(400MHz,CDCl
3)δ9.09(s,1H),8.73(s,1H),8.16(s,1H),7.65-7.58(m,1H),7.56-7.50(m,4H),7.47-7.39(m,4H),7.34-7.29(m,2H),7.26-7.21(m,1H),6.85(d,J＝8.3Hz,4H),6.31(dd,J＝7.0,11.4Hz,1H),5.72-5.51(m,1H),5.46-5.34(m,1H),4.69(br d,J＝5.1Hz,1H),3.79(s,3H),3.78(s,3H),3.53-3.44(m,1H),3.37(s,1H),3.03(br t,J＝12.1Hz,1H).
步骤5：化合物1-6的制备
氮气保护下，将2-氰乙基-N,N-二异丙基氯代亚磷酰胺(525mg,2.22mmol)滴加入化合物1-5(1g,1.48mmol)和二异丙基乙基胺(742.00mg,5.74mmol,1.00mL)的乙腈(10mL)溶液中，加毕，反应体系室温搅拌反应2h。停止反应，加入乙酸乙酯(100mL)稀释，碳酸氢钠饱和溶液(50mL x 3)洗涤，有机相用饱和食盐水(50mL x 2)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，化合物1-6。
MS(ESI)m/z＝793.4.
1H NMR(400MHz,CDCl
3)δ9.04(br d,J＝12.0Hz,1H),8.76(d,J＝4.6Hz,1H),8.23(d,J＝16.9Hz,1H),8.01(br d,J＝7.3Hz,2H),7.65-7.56(m,1H),7.55-7.48(m,4H),7.44-7.37(m,4H),7.31-7.26(m,2H),7.24-7.20(m,1H),6.83-6.77(m,4H),6.44-6.19(m,1H),4.64-4.50(m,1H),4.31-4.00(m,2H),3.76(dd,J＝3.7,8.1Hz,6H),3.65-3.29(m,5H),2.80-2.71(m,1H),2.67-2.59(m,1H),2.53(t,J＝6.5Hz,1H),1.13(dd,J＝3.4,6.6Hz,6H),1.05(d,J＝6.8Hz,3H),0.99(d,J＝6.8Hz,3H).
31P NMR(162MHz,CDCl
3)δ148.91,148.74.
步骤6：化合物1-8的制备
将化合物1-7(10g,26.93mmol)溶于吡啶(150mL)中，滴加入4,4'-双甲氧基三苯甲基氯(11.86g,35.01mmol)，室温反应16h。反应加水(100mL)淬灭，体系加入二氯甲烷(200mL)，滤除固体，分出有机相，水(100mL x 4)洗，无水硫酸钠干燥，过滤，滤液减压浓缩，所得固体经硅胶柱层析纯化(洗脱剂：石油醚/乙酸乙酯(v/v)＝20/1～10/1；二氯甲烷/甲醇(v/v)＝1/0～10/1)，得粗品，粗品溶于二氯甲烷(50mL)中，然后溶液滴加入甲基叔丁基醚(200mL)中，滤出固体，真空干燥，得化合物1-8。
MS(ESI)m/z(M+H)
+＝674.3.
1H NMR(400MHz,DMSO-d
6)δ11.25(s,1H),8.69(s,1H),8.60(s,1H),8.04(d,J＝7.6Hz,2H),7.69-7.59(m,1H),7.59-7.51(m,2H),7.36(d,J＝7.6Hz,2H),7.28-7.15(m,7H),6.88–6.76(m,4H),6.07(d,J＝4.8Hz,1H),5.76(s,1H),5.67(d,J＝5.6Hz,1H),5.31(d,J＝5.6Hz,1H),4.84-4.72(m,1H),4.40-4.47(m,1H),3.71(d,J＝1.2Hz,6H),3.23(d,J＝4.8Hz,2H).
步骤7：化合物1-9B的制备
在氮气保护下，将化合物1-8(15g,22.26mmol)和咪唑(4.55g,66.79mmol)溶于吡啶(60mL)中，加入叔 丁基二甲基氯硅烷(5.03g,33.40mmol,4.09mL)，反应体系室温搅拌4h。体系中加入乙酸乙酯(200mL)稀释，滤除白色固体，溶液浓缩至干，复溶于乙酸乙酯(200mL)中，饱和食盐水(100mL x 4)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～5/3)，得化合物1-9A(6.00g,产率：29.3％，第三个峰)，化合物1-9B(4.00g,产率：22.3％，第二个峰)，化合物1-9C(5.60g,产率：31.4％，第一个峰)。
化合物1-9A:
MS(ESI)m/z(M+H)
+＝788.4.
1H NMR(400MHz,CDCl
3)δ9.03(br s,1H),8.76(s,1H),8.25(s,1H),8.01(br d,J＝7.3Hz,2H),7.63-7.55(m,1H),7.55-7.47(m,2H),7.37(br d,J＝7.1Hz,2H),7.30-7.16(m,8H),7.20-7.11(m,1H),6.78(br d,J＝8.8Hz,4H),6.06(d,J＝4.9Hz,1H),4.78–4.74(m,1H),4.62-4.55(m,1H),4.18(br d,J＝3.9Hz,1H),3.76(s,6H),3.51(dd,J＝3.3,10.6Hz,1H),3.29-3.16(m,2H),0.88(s,9H),0.08(s,3H),0.00(s,3H).
化合物1-9B:
MS(ESI)m/z(M+H)
+＝788.4.
1H NMR(400MHz,CDCl
3)δ9.23(s,1H),8.88(s,1H),8.38(s,1H),8.17(br d,J＝7.3Hz,2H),7.79-7.70(m,1H),7.67(t,J＝7.6Hz,2H),7.59(br d,J＝7.6Hz,2H),7.48(br d,J＝8.6Hz,4H),7.44-7.30(m,4H),6.96(br d,J＝8.8Hz,4H),6.25(d,J＝5.1Hz,1H),5.19-5.11(m,1H),4.55-4.47(m,1H),4.40-4.45(m,1H),3.92(s,6H),3.73-3.64(m,1H),3.52-3.55(m,1H),2.87(d,J＝3.9Hz,1H),0.98(s,9H),0.14(s,3H),0.00(s,3H).
步骤8：化合物1-10的制备
氮气保护下，将化合物1-6(1.3g,1.48mmol)的乙腈(5mL)溶液滴加入化合物1-9B(1.16g,1.47mmol)、四氮唑(0.45M的乙腈溶液,32.98mL)和
分子筛(2g)的乙腈(20mL)溶液中，室温反应，4h后，加入(E)-N,N-二甲基-N'-(3-硫代-3H-1,2,4-二硫唑-5-基)甲脒(1g,4.87mmol)，继续搅拌1h。反应体系过滤，滤液用乙酸乙酯(100mL)稀释，有机相依次用饱和碳酸氢钠溶液(50mL x 3)、饱和食盐水(50mL x 3)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/乙酸乙酯(v/v)＝9/1)，得化合物1-10。MS(ESI)m/z(M/2+H)
+＝798.4.
步骤9：化合物1-11的制备
将化合物1-10(2.1g,1.32mmol)溶于二氯甲烷(10mL)中，滴加入二氯乙酸(24.04g,5.27mmol,18mL,5％的二氯甲烷溶液)和三乙基硅烷(10.92g,93.91mmol,15mL)，反应体系室温搅拌反应1.5h。反应体系加入二氯甲烷(100mL)，依次用水(100mL)、饱和碳酸氢钠(100mL x 2)、饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，所得固体经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝20/1)，得化合物1-11。MS(ESI)m/z(M+H)
+＝990.3.
1H NMR(400MHz,CDCl
3)δ9.24(br s,1H),8.84-8.74(m,2H),8.32-8.19(m,2H),8.07-7.99(m,3H),7.89-7.84(m,1H),7.66-7.47(m,6H),7.44-7.37(m,1H),6.39-6.26(m,1H),5.98(d,J＝7.6Hz,0.5H),5.79(d,J＝7.8Hz,0.5H),5.73(t,J＝5.3Hz,0.5H),5.60(t,J＝5.3Hz,0.5H),5.25-4.94(m,3H),4.63-4.21(m,6H),4.03-3.92(m,1H),3.83-3.72(m,1H),3.70-3.62(m,1H),2.84-2.73(m,3H),0.69(d,J＝2.4Hz,9H),-0.11--0.21(m,3H),-0.32--0.45(m,3H).
31P NMR(162MHz,CDCl
3)δ68.00,67.97.
步骤10：化合物1-12的制备
氮气保护下，将化合物1-11(1g,1.01mmol)、
分子筛(2g)和四氮唑(0.45M的乙腈溶液,58mL)与乙腈(15mL)混合，反应体系滴加入2-氰乙基N,N,N',N'-四异丙基亚磷酰二胺(431.36mg,1.43mmol,454.55μL)的乙腈(10mL)溶液，30min加完，反应体系室温搅拌反应1h。停止反应，过滤，滤液中加入乙酸乙酯(150mL)稀释，有机相依次用碳酸氢钠饱和溶液(100mL x 3)、饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，所得固体经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝20/3)，得化合物1-12。
MS(ESI)m/z(M/2+H)
+＝545.6.
步骤11：化合物1-13的制备
氮气保护、0℃条件下，将硼烷二甲硫醚(2M的二氯甲烷溶液,1.00mL)滴加入化合物1-12(500mg,459.11μmol)、
分子筛(500mg)的二氯甲烷(15mL)溶液中，加毕升温至15℃反应20min。停止反应，加水(5mL)淬灭反应，二氯甲烷(40mL)稀释，过滤，滤液用水(30mL x 3)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，得化合物1-13，无需进一步纯化直接用于下一步反应。
步骤12：化合物1-14的制备
将化合物1-13(480mg,435.22μmol)溶于30％的甲胺乙醇溶液(15mL)中，反应室温搅拌反应12h。反应体系减压浓缩，所得固体溶于水(20mL)中，用乙酸乙酯(10mL)萃取，水相冻干得化合物1-14。
步骤13：化合物1-14A、1-14B、1-14C和1-14D的制备
化合物1-14粗品溶于水(10mL)中，高效制备液相分离(分离条件:色谱柱:Xbridge 150*30mm*10μm；流动相:[水(10mM碳酸氢铵)-乙腈]；乙腈％:12％-32％,流速：25mL/min，7min)。
化合物1-14A(HPLC保留时间3.283min)
化合物1-14B(HPLC保留时间3.654min)
化合物1-14C(HPLC保留时间4.282min)
化合物1-14D(HPLC保留时间4.866min)
化合物1-14A:
MS(ESI)m/z(M+H)
+＝789.2.
1H NMR(400MHz,DMSO-d
6)δ8.64(s,1H),8.52(s,1H),8.24(s,1H),8.22(s,1H),8.11-7.41(m,4H),6.37-6.27(m,1H),5.96(s,1H),5.25-5.02(m,1H),4.98-4.62(m,3H),4.41-4.18(m,4H),3.98-3.80(m,2H),0.94(m,9H),0.51--0.16(m,9H).
31P NMR(162MHz,DMSO-d
6)δ92.31-89.90,52.80.
19F NMR(376MHz,DMSO-d
6)δ-201.02.
化合物1-14B:
MS(ESI)m/z(M+H)
+＝789.3.
1H NMR(400MHz,DMSO-d
6)δ8.62-8.50(m,1H),8.44-8.33(m,1H),8.24(br s,1H),8.19(s,1H),8.13-7.38(m,4H),6.38-6.23(m,1H),5.96(s,1H),5.47-5.14(m,1H),4.91-4.68(m,2H),4.51-4.17(m,5H),3.87-3.69(m,2H),0.95(s,9H),0.25(s,3H),0.24(s,3H),0.13--0.44(m,3H).
31P NMR(162MHz,DMSO-d
6)δ91.24-90.10,53.03.
19F NMR(376MHz,DMSO-d
6)δ-201.36.
化合物1-14C:
MS(ESI)m/z 775.5.
1H NMR(400MHz,DMSO-d
6)δ8.98(br s,1H),8.78(br s,1H),8.59-8.39(m,1H),8.29-8.05(m,2H),7.82(br s,2H),6.34(br d,J＝12.8Hz,1H),6.13-5.89(m,1H),5.58-5.32(m,1H),5.17(br s,1H),5.05-4.64(m,3H),4.45-4.22(m,3H),3.84-3.74(m,2H),1.00(s,9H),0.76--0.11(m,9H).
31P NMR(162MHz,DMSO-d
6)δ92.51-90.91,50.69.
19F NMR(376MHz,DMSO-d
6)δ-201.87.
化合物1-14D:
MS(ESI)m/z 775.3.
1H NMR(400MHz,DMSO-d
6)δ9.26(s,1H),8.97(s,1H),8.82-8.37(m,4H),8.29(s,1H),7.82(s,1H),6.46-6.31(m,1H),6.16(s,1H),6.11-5.99(m,1H),5.77-5.57(m,1H),5.13-5.01(m,2H),4.64(br d,J＝11.5Hz,1H),4.55-4.37(m,2H),3.93-3.79(m,2H),1.14(s,9H),0.95(m,6H),0.74(br s,3H).
31P NMR(162MHz,DMSO-d
6)δ91.98-90.66,52.69.
19F NMR(376MHz,DMSO-d
6)δ-201.92.
步骤14：化合物1A的制备
将光活性异构体化合物1-14A(20mg,24.31μmol)溶于吡啶(2mL)中，依次加入三乙胺(290.80mg,2.87mmol,0.4mL)和三乙胺三氢氟酸盐(197.80mg,1.23mmol,0.2mL)，反应体系升温至50℃搅拌反应14h。冷至室温，加入异丙氧基三甲基硅烷(745mg,5.63mmol,1mL)，继续室温反应4h。反应减压浓缩，浓缩残余物溶于水(2mL)，乙酸乙酯(3mL)反萃，水相用高效液相制备分离(分离条件:色谱柱:Xbridge 150*30mm*10μm；流动相:[水(10mM碳酸氢铵)-乙腈]；乙腈％：0％-20％,流速：25mL/min,7min)，得化合物1A(HPLC保留时间1.386min)。
MS(ESI)m/z(M+H)
+＝674.8.
1H NMR(400MHz,D
2O)δ8.42(br s,2H),8.18(br s,1H),8.06(br s,1H),6.37(br d,J＝15.8Hz,1H),6.13(s,1H),5.62-5.36(m,1H),5.05(br s,1H),4.95-4.77(m,2H),4.46(br t,J＝7.3Hz,2H),4.40-4.26(m,2H),4.08 -3.93(m,2H),0.10(br s,3H).
31P NMR(162MHz,D
2O)δ93.99-91.82,54.64.
19F NMR(376MHz,D
2O)δ-202.66.
步骤15：化合物1B,1C,1D的制备
其他光活性纯异构体1B,1C,1D可分别由化合物1-14B,1-14C,1-14D参考化合物1A制备方法制得。化合物1B：
MS(ESI)m/z(M-H)
-＝672.9.
1H NMR(400MHz,D
2O)δ8.10(br s,1H),8.06(br s,1H),7.95(br s,1H),7.66(br s,1H),6.34-6.16(m,1H),5.91(br d,J＝5.9Hz,1H),5.54-5.25(m,1H),4.55-4.34(m,5H),4.29(br d,J＝7.3Hz,1H),4.18(br d,J＝12.2Hz,1H),3.92-3.80(m,2H),0.08(br s,3H).
31P NMR(162MHz,D
2O)δ93.84-92.90,53.89.
19F NMR(376MHz,D
2O)δ-203.40.
化合物1C：
MS(ESI)m/z(M-H)
-＝672.9.
1H NMR(400MHz,D
2O)8.30-7.95(m,3H),7.75(br s,1H),6.24(d,J＝13.8Hz,1H),6.08(s,1H),5.35-5.06(m,1H),4.97(br d,J＝3.3Hz,1H),4.57-4.28(m,6H),4.02-3.87(m,2H),0.32(br s,3H).
31P NMR(162MHz,D
2O)δ95.59-93.68,53.85.
19F NMR(376MHz,D
2O)δ-202.82.
化合物1D：
MS(ESI)m/z(M-H)
-＝672.9.
1H NMR(400MHz,D
2O)δ7.97(br s,1H),7.83-7.75(m,3H),6.23(br d,J＝11.2Hz,1H),5.94(br s,1H),5.24-4.97(m,1H),4.44-4.17(m,7H),3.75(br s,2H),0.18(br s,3H).
31P NMR(162MHz,D
2O)δ94.34-93.26,53.47.
19F NMR(376MHz,D
2O)δ-203.12.
实施例2：化合物2A、2B的制备
步骤1：化合物2-2的制备
氩气保护下，将化合物1-5(1.8g,2.66mmol)、
分子筛(2g)和四氮唑(0.45M乙腈溶液,88.80mL)分散于乙腈(10mL)中。室温搅拌10分钟后加入化合物2-1(2.33g,2.66mmol)的乙腈(10mL)溶液。反应室温搅拌1小时，反应液用乙酸乙酯(50mL)稀释后过滤。滤液用饱和碳酸氢钠溶液(40mL x 3)和饱和食盐水(10mL)洗涤，无水硫酸镁干燥后，真空浓缩旋干得到化合物2-2。
MS(ESI)m/z(M/2+H)
+＝725.8.
步骤2：化合物2-3的制备
在0℃下，将BH
3-Me
2S(2M的四氢呋喃溶液,3.31mL)慢慢加入到化合物2-2(3.2g,2.21mmol)、
分子筛(3g)和二氯甲烷(35mL)的混合溶液中。反应室温搅拌40min后过滤。滤饼用乙酸乙酯(50mL)洗涤。水(20mL)加入到滤液中，然后用乙酸乙酯(20mL x 3)萃取。合并的有机相用Na
2SO
4干燥，减压浓 缩得到化合物2-3，粗品直接用到下一步反应。
步骤3：化合物2-4的制备
将化合物2-3(3.2g,2.19mmol)溶解在乙腈(9mL)和乙酸(80％的水溶液,27mL)的混合溶液中，反应液室温搅拌18小时。加入乙酸乙酯(20mL)稀释后，用饱和碳酸氢钠(10mL x 3)和饱和食盐水(5mL)洗涤。有机相用硫酸钠干燥，过滤，滤液真空浓缩，残留物用柱层析分离(洗脱剂：石油醚/乙酸乙酯＝0～100％，然后二氯甲烷/甲醇＝0～10％)纯化得到化合物2-4。
MS(ESI)m/z(M+H)
+＝860.3.
1H NMR(400MHz,DMSO-d
6)δ11.25(s,H),11.23(s,1H),8.73-8.66(m,2H),8.57(s,1H),8.56(s,1H),8.08-7.96(m,4H),7.72-7.43(m,6H),6.50-6.32(m,2H),6.06-5.77(m,2H),5.70-5.23(m,3H),4.90-4.66(m,1H),4.52-4.30(m,2H),4.26-4.06(m,4H),3.73-3.52(m,2H),2.99-2.79(m,2H),0.68-0.15(br,3H).
31P NMR(162MHz,DMSO-d
6)δ114.2-115.2
19F NMR(376MHz,DMSO-d
6)δ-201.38--201.75,-204.16--204.3
步骤4：化合物2-5的制备
氩气保护下，将
分子筛(0.5g)加入到化合物2-4(200mg,232.68μmol)的乙腈(1mL)溶液中，在室温下加入四氮唑(0.45M乙腈溶液,7.76mL)。混合物室温搅拌15min后逐批加入2-氰乙基N,N,N',N'-四异丙基亚磷酰二胺(105.20mg,349.02μmol)。反应液在室温条件下搅拌1小时。过滤除去分子筛，滤饼用乙酸乙酯(5mL)洗涤三次。滤液用饱和碳酸氢钠溶液(5mL x 3)和饱和食盐水(5mL)洗涤后用无水硫酸钠干燥，真空浓缩旋干得到化合物2-5。产物不经纯化，直接用于下一步反应。
MS(ESI)m/z(M+H)
+＝959.4.
步骤5：化合物2-6的制备
在0℃下，硼烷二甲硫醚(2M的四氢呋喃溶液,344.26μL)逐滴加入到化合物2-5(220mg,229.51μmol)的二氯甲烷(6mL)溶液中。反应液在0℃下搅拌20分钟，加入水(2mL)淬灭。继续搅拌10分钟后用二氯甲烷(5mL x 3)萃取。有机相用饱和食盐水(5mL)洗涤后，无水硫酸钠干燥，真空旋干得到化合物2-6。产物不经纯化，直接用于下一步反应。
步骤6：化合物2A，2B和2C的制备
将化合物2-6(0.5g,0.502mol)溶于30％的甲胺乙醇溶液(20mL)中，反应于35℃搅拌反应72h。反应体系减压浓缩，所得固体经高效制备液相分离(分离条件：色谱柱：Waters Xbridge Prep OBD C18 150*30mm*10μm；流动相：[水(10mM碳酸氢铵)-乙腈]；乙腈％:20％-90％，流速：25mL/min，20min)。
化合物2A(HPLC保留时间:t＝9.2min)。
MS(ESI)m/z(M-H)
-＝657.3.
1H NMR(400MHz,D
2O)δ8.21(s,2H),7.88(s,2H),6.26(s,1H),6.22(s,1H),5.50(s,1H),5.37(s,1H),4.89-4.73(m,2H),4.38-4.35(m,2H),4.18-4.15(m,2H),3.92-3.85(m,2H),0.45--0.2(br,6H).
31P NMR(162MHz,D
2O)δ93.43-92.29
19F NMR(376MHz,D
2O)δ-203.09,
化合物2B(HPLC保留时间:t＝11.1min)。
MS(ESI)m/z(M+H)
+＝659.2.
1H NMR(400MHz,D
2O)δ8.16(s,1H),8.03(s,2H),7.62-7.75(m,1H),6.35-6.30(m,1H),6.18-6.14(m,2H),5.42-5.25(m,2H),5.19-5.07(m,.2H),4.48-4.35(m,2H),4.36-4.26(m,2H),4.26-4.17(m,2H),3.85-3.81(m,2H),0.55--0.15(br,6H)
31P NMR(162MHz,D
2O)δ92.77-90.51
19F NMR(376MHz,D
2O)δ-202.63
化合物2C(HPLC保留时间:t＝17.0min)。
MS(ESI)m/z(M+H)
+＝659.4.
1H NMR(400MHz,D
2O)δ8.12(s,2H),7.84(s,2H),6.15(s,1H),6.11(s,1H),5.42-5.26(m,2H),5.09-5.03(m,2H),4.35-4.28(m,4H),3.89-3.84(m,2H),0.55--0.1(br,6H)
31P NMR(162MHz,D
2O)δ95.02-92.41
19F NMR(376MHz,D
2O)δ-201.85
步骤7：化合物2D的制备
Dowex-50W离子交换树脂(20g)置于烧杯中，用去离子水(10mL)洗涤，然后加入硫酸(15％的去离子水溶液)，搅拌5min，倾出液体，将树脂转移至色谱柱中，依次用硫酸(15％的去离子水溶液，4CV)和去离子水冲洗至pH＝7.0。处理后的树脂移入烧杯中，加入氢氧化钠(15％的去离子水溶液)，搅拌5min，倾出液体，树脂转入色谱柱，依次用氢氧化钠(15％的去离子水溶液)和水冲洗至pH＝7.0。化合物2B(140mg,202.28umol,2NH
4)溶于水(2mL)中，经上述所得色谱柱纯化得化合物2D。
MS(ESI)m/z(M+H)
+＝659.3.
1H NMR(400MHz,D
2O)δ8.27(s,1H),8.23(s,1H),8.05(s,1H),7.78(s,1H),6.33-6.25(m,2H),5.55-5.42(m,2H),5.05-4.90(m,2H),4.44-4.36(m,3H),4.29-4.25(m,1H),4.02-3.96(m,2H),0.55--0.2(br,6H).
31P NMR(162MHz,D
2O)δ93.43-92.29
19F NMR(376MHz,D
2O)δ-203.09
实施例3：化合物3A、3B、3C、3D的制备
步骤1：化合物3-2的制备
氩气保护下，化合物4-氯-5-氟-7H-吡咯并[2,3-D]-嘧啶(1.03g,6.0mmol)溶于乙腈(40mL)溶液中，加入BSA(1.76mL,7.2mmol)，反应体系搅拌5min后依次加入3-1(3.0g,6.0mmol)和三氟甲磺酸三甲基硅酯(1.32mL,7.2mmol)。25℃反应30min后升温至80℃反应3h。加入水(100mL)淬灭反应，乙酸乙酯(100 mL x 3)萃取，合并有机相，饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝5/1)，得化合物3-2.
MS(ESI)m/z(M+H)
+＝616.1
1H NMR(400MHz,CDCl
3)δ8.59(s,1H),8.11(d,J＝8.0Hz,2H),8.01(d,J＝8.0Hz,2H),7.91(d,J＝8.0Hz,2H),7.65-7.38(m,9H),7.18(s,1H),6.69(d,J＝8Hz,1H),6.15-6.07(m,2H),4.91-4.66(m,3H).
步骤2：化合物3-3的制备
氩气保护下，化合物3-2(200mg,0.32mmol)加入到甲醇钠的甲醇溶液(4mL,0.5mol/L,1.92mmol)中。25℃反应1h。反应体系加入醋酸调节pH到7.0，浓缩得粗品，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝10/1)，得化合物3-3.
MS(ESI)m/z(M+H)
+＝300.1
1H NMR(400MHz,DMSO-d
6)δ8.45(s,1H),7.66(s,1H),6.18(d,J＝4.0Hz,1H),5.35(d,J＝4.0Hz,1H),5.16(d,J＝4.0Hz,1H),5.10-5.06(m,1H),4.35-4.31(m,1H),4.11-4.09(m,1H),4.07(s,3H),3.92-3.88(m,1H),3.65-3.51(m,2H).
步骤3：化合物3-4的制备
氩气保护下，化合物3-3(100mg,0.34mmol)溶于乙腈(10mL)中，依次加入碘化钠(250mg,1.68mmol)和三甲基碘硅烷(0.2mL,1.56mmol)。25℃反应搅拌3h，减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝8/1)，得化合物3-4.
MS(ESI)m/z(M+H)
+＝286.0
1H NMR(400MHz,DMSO-d
6)δ12.1(s,1H),7.92(d,J＝3.2Hz,1H),7.35(d,J＝3.2Hz,1H),6.06(dd,J＝8.0Hz,3.2Hz,1H),5.35-5.02(m,3H),4.23(s,1H),4.06-4.04(m,1H),3.88-3.86(m,1H),3.62-3.51(m,2H).
步骤4：化合物3-5的制备
氮气保护下，DMTrCl(2.57g,7.57mmol)缓慢加入到化合物3-4(1.8g,6.31mmol)的吡啶(10mL)溶液中，反应体系25℃搅拌反应12h。减压浓缩至干，复溶于乙酸乙酯(50mL)中，有机相用饱和食盐水(20 mL x 5)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝20/1)，得化合物3-5.
MS(ESI)m/z(M+H)
+＝588.3.
1H NMR(400MHz,DMSO-d
6)12.15(br s,1H),7.93(d,J＝2.9Hz,1H),7.41-7.34(m,2H),7.32-7.14(m,8H),6.86(dd,J＝2.0,8.8Hz,4H),6.08(d,J＝3.7Hz,1H),5.50(d,J＝5.9Hz,1H),5.19(d,J＝5.6Hz,1H),4.34-4.25(m,1H),4.12(q,J＝5.1Hz,2H),3.99(q,J＝4.5Hz,1H),3.73(s,6H).
步骤5：化合物3-6的制备
氮气保护下，化合物3-5(0.85g,1.45mmol)溶于吡啶(8mL)中，依次加入咪唑(200mg,2.94mmol)和叔丁基二甲基氯硅烷(265mg,1.76mmol)，反应体系25℃搅拌反应12h。减压除去大部分溶剂，反应液倒入乙酸乙酯(30mL)中，有机相用饱和食盐水(10mL x 3)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝2/1)，得化合物3-6.
MS(ESI)m/z(M+H)
+＝702.3.
1H NMR(400MHz,DMSO-d
6)12.14(br s,1H),7.92(br s,1H),7.39(br s,2H),7.33-7.21(m,7H),7.13(br s,1H),6.88(br s,4H),6.11(br s,1H),5.10(br s,1H),4.40(br d,J＝4.4Hz,1H),4.16-3.94(m,3H),3.78-3.68(m,6H),3.23(br d,J＝9.3Hz,1H),0.79-0.70(m,9H),-0.03(br d,J＝2.2Hz,3H),-0.10--0.17(m,3H).
步骤6：化合物3-7的制备
化合物1-6(0.7g,997.36μmol)溶于四氢呋喃(4mL)中，依次加入
分子筛(1g)和四氮唑(0.45M的乙腈溶液,33.25mL)，然后25℃氩气保护条件下，向反应体系中滴加入化合物3-6(1.05g,1.20mmol)的乙腈(6mL)溶液，搅拌反应1h。反应液用乙酸乙酯(50mL)稀释，过滤，滤液依次用碳酸氢钠饱和溶液(50mL x 3)和饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经快速柱层析纯化(二氯甲烷/甲醇(v/v)＝10/1)，得化合物3-7.
31P NMR(162MHz,CDCl
3)δ138.84,138.40.
19F NMR(376MHz,CDCl
3)δ-163.24,-196.85--198.69.
步骤7：化合物3-8的制备
0℃，氩气保护下，化合物3-7(1.8g,1.22mmol)溶于二氯甲烷(30mL)中，依次加入
分子筛(2g)和硼烷二甲硫醚络合物(2M的四氢呋喃溶液,2.44mL)，升温至20℃搅拌反应30min。反应用水(5mL)淬灭，二氯甲烷(40mL)稀释，过滤，滤液用水(30mL x 3)洗涤，有机相用无水硫酸钠干燥，过滤，滤液减压浓缩，得粗品3-8，不经进一步纯化直接用于下一步反应。
步骤8：化合物3-9的制备
化合物3-8(1.6g,1.07mmol)溶于二氯甲烷(20mL)中，依次加入2,2-二氯乙酸(1.23g,5.37mmol,5％的二氯甲烷溶液)和三乙基硅烷(14.56g,125.22mmol,20mL)，加毕，反应体系25℃搅拌反应1h。反应加二氯甲烷(50mL)稀释，有机相依次用水(50mL)、饱和碳酸氢钠溶液(50mL x 2)和饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品依次经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝9/1)，高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*30mm 5μm；流动相:[水(10mM碳酸氢铵)-乙腈]；乙腈％:45％-45％,流速：25mL/min，8min)，得化合物3-9(HPLC保留时间3.488min)。
MS(ESI)m/z(M+H)
+＝886.2
步骤8：化合物3-10的制备
20℃，氩气保护条件下，化合物3-9(180mg,203.23μmol)溶于乙腈(2mL)中，依次加入
分子筛 (0.5g)和1H-四氮唑的乙腈溶液(0.45M,9.03mL)，滴加入2-氰乙基N,N,N',N'-四异丙基亚磷酰二胺(91.88mg,304.85μmol,96.82μL)，反应体系搅拌反应1h。过滤，滤液用乙酸乙酯(30mL)稀释，有机相用饱和碳酸氢钠溶液(20mL x 2)洗涤，无水硫酸钠干燥，减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝15/1)，得化合物3-10。
31P NMR(162MHz,CD
3CN)δ139.33,138.84,137.41,136.85.
19F NMR(376MHz,CD
3CN)δ-165.95,166.02,-199.48--201.40.
步骤9：化合物3-11的制备
0℃，氩气保护条件下，硼烷二甲硫醚络合物(2M的四氢呋喃溶液,243.72μL)滴加入化合物3-10(160mg,162.48μmol)和4A分子筛(200mg)的二氯甲烷(5mL)溶液中，加毕，反应体系15℃条件下搅拌30min。加水(5mL)淬灭反应，加二氯甲烷(40mL)稀释，过滤，滤液用水(30mL)洗涤，有机相用无水硫酸钠干燥，过滤，滤液浓缩，得粗品3-11，不经进一步纯化直接用于下一步反应。
步骤10：化合物3-12A，3-12B，3-12C，3-12D的制备
将化合物3-11(140mg,140.20μmol)溶于30％的甲胺乙醇溶液(5mL)中，反应20℃条件下搅拌反应72h。反应体系减压浓缩，所得残渣溶于水(10mL)中，乙酸乙酯(10mL)反萃，水相冻干，所得粗品经高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*30mm 5μm；流动相:[水(0.04％氨水+10mM碳酸氢铵)-乙腈]；乙腈％:5％-45％,流速：25mL/min，7min)。得：
化合物3-12A(HPLC保留时间2.143min)
化合物3-12B和3-12C的混合物(HPLC保留时间2.304min)
化合物3-12D(HPLC保留时间2.596min)
化合物3-12A：
MS(ESI)m/z(M+H)
+＝789.3.
1H NMR(400MHz,CD
3OD)δ8.74(s,1H),8.29(s,1H),7.84(d,J＝1.5Hz,1H),7.59(s,1H),6.40(d,J＝15.4Hz,1H),6.21(s,1H),5.62-5.40(m,1H),5.31-5.08(m,1H),4.55-4.24(m,6H),4.03-3.97(m,2H),0.99(s,9H),0.83--0.35(m,12H).
31P NMR(162MHz,DMSO-d
6)δ93.38-90.01.
19F NMR(376MHz,DMSO-d
6)δ-165.43,-200.43--200.57.
化合物3-12B和3-12C的混合物：
MS(ESI)m/z(M+H)
+＝789.3.
31P NMR(162MHz,DMSO-d
6)δ93.82-90.37.
19F NMR(376MHz,DMSO-d
6)δ-164.75,-165.44,-199.67--200.85.
化合物3-12D：
MS(ESI)m/z(M+H)
+＝789.3.
1H NMR(400MHz,CD
3OD)δ8.62(br s,1H),8.21(br s,1H),7.86-7.61(m,1H),7.42(br s,1H),6.37(br d,J＝16.6Hz,1H),6.07(br s,1H),5.58-5.32(m,1H),5.28-5.08(m,1H),4.59-4.26(m,6H),4.02-3.80(m,2H),1.07-0.85(m,9H),0.73-0.05(m,12H).
31P NMR(162MHz,DMSO-d
6)δ94.76-91.59.
19F NMR(376MHz,DMSO-d
6)δ-165.15,-200.66--200.94.
步骤11：化合物3A的制备
化合物3-12A(10mg,12.69μmol)溶于吡啶(1mL)中，依次加入三乙胺(73.83mg,729.59μmol,101.55μL)和三乙胺三氟化氢盐(49.01mg,304.00μmol,49.55μL)，反应升温至50℃搅拌反应48h。体系冷至25℃，然后加入异丙氧基三甲基硅烷(193.01mg,1.46mmol,259.08μL)，反应体系25℃搅拌4h。减压浓缩，残渣溶于水(3mL)中，乙酸乙酯(3mL)反萃，收集水相，经高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*30mm 5μm；流动相:[水(10mM碳酸氢铵)-乙腈]；乙腈％:0％-30％,流速：25mL/min，7min)。得化合物3A。
MS(ESI)m/z(M-H)
+＝672.8.
1H NMR(400MHz,D
2O)δ8.23(br s,1H),7.95(br s,1H),7.77(br s,1H),7.08(br s,1H),6.23(br d,J＝16.1Hz,1H),6.05(br s,1H),5.64-5.36(m,1H),5.01-4.74(m,1H),4.57(br s,1H),4.44-4.30(m,2H),4.20(br d,J＝8.8Hz,1H),4.15-4.03(m,2H),3.84(t,J＝12.2Hz,2H),0.02(br s,6H).
31P NMR(162MHz,D
2O)δ94.63-92.15.
19F NMR(376MHz,D
2O)δ-165.60,-203.17--203.42.
步骤12：化合物3B和3C的制备
化合物3-12B和3-12C的混合物(20mg,24.32μmol,2NH
4
+)溶于吡啶(1mL)中，依次加入三乙胺(147.65mg,1.46mmol,203.10μL)和三乙胺三氟化氢盐(117.62mg,729.59μmol,118.92μL)，反应升温至50℃搅拌反应48h。体系冷至25℃，然后加入异丙氧基三甲基硅烷(386.03mg,2.92mmol,518.16μL)，反应体系25℃搅拌4h。减压浓缩，残渣溶于水(3mL)中，乙酸乙酯(3mL)反萃，收集水相，经高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*30mm 5μm；流动相:[水(0.04％氨水+10mM碳酸氢铵)-乙腈]；乙腈％:0％-30％,流速：25mL/min，7min)。得化合物3B(HPLC保留时间6.24min)和化合物3C(HPLC保留时间6.27min)。
化合物3B：
MS(ESI)m/z(M-H)
-＝672.9.
1H NMR(400MHz,D
2O)δ8.29(s,1H),7.97(s,1H),7.78(s,1H),7.08(d,J＝1.8Hz,1H),6.25(d,J＝16.6Hz,1H),6.07(s,1H),5.42-5.17(m,1H),5.14-4.97(m,1H),4.64-4.58(m,1H),4.36-4.26(m,2H),4.13(br d,J＝10.0Hz,3H),3.86(dd,J＝5.3,12.0Hz,1H),3.78(br dd,J＝5.1,11.2Hz,1H),0.00(br s,3H),-0.17(br s,3H).
31P NMR(162MHz,D
2O)δ94.34-93.23.
19F NMR(376MHz,D
2O)δ-164.48,-201.40.
化合物3C：
MS(ESI)m/z(M-H)
-＝672.9.
1H NMR(400MHz,D
2O)δ8.30(br s,1H),8.05(br s,1H),7.77(br s,1H),7.12(br s,1H),6.27(br d,J＝15.4Hz,1H),6.09(br s,1H),5.62-5.23(m,1H),4.92-4.69(m,3H),4.40-4.23(m,3H),4.18(br d,J＝9.0Hz,1H),4.07(br d,J＝11.7Hz,1H),3.86(br d,J＝8.3Hz,2H),0.06(br s,6H).
31P NMR(162MHz,D
2O)δ93.82-89.83.
19F NMR(376MHz,D
2O)δ-165.87,-202.61--203.31.
步骤13：化合物3D的制备
化合物3-12D(10mg,12.16μmol,2NH
4
+)溶于吡啶(1mL)中，依次加入三乙胺(73.83mg,729.59μmol,101.55μL)和三乙胺三氟化氢盐(49.01mg,304.00μmol,49.55μL)，反应升温至50℃搅拌反应48h。体系冷 至25℃，然后加入异丙氧基三甲基硅烷(193.01mg,1.46mmol,259.08μL)，反应体系25℃搅拌4h。减压浓缩，残渣溶于水(3mL)中，乙酸乙酯(3mL)反萃，收集水相，经高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*30mm 5μm；流动相:[水(0.04％氨水+10mM碳酸氢铵)-乙腈]；乙腈％:0％-30％,流速：25mL/min，7min)。得化合物3D。
MS(ESI)m/z(M-H)
-＝672.8.
1H NMR(400MHz,D
2O)δ8.30(br s,1H),8.12(br s,1H),7.69(br s,1H),7.15(br s,1H),6.29(br dd,J＝3.3,16.1Hz,1H),6.04(br d,J＝7.0Hz,1H),5.76-5.50(m,1H),5.24-5.09(m,1H),4.97-4.92(m,1H),4.49-4.43(m,2H),4.40(br d,J＝12.0Hz,1H),4.31(br d,J＝8.0Hz,2H),4.02-3.85(m,2H),0.29(br s,6H).
31P NMR(162MHz,D
2O)δ94.73-93.43.
19F NMR(376MHz,D
2O)δ-164.94,-201.79.
实施例4：化合物4A、4B、4C、4D的制备
步骤1：化合物4-1的制备
氩气保护，15℃条件下，化合物2-2(2.3g,1.59mmol)溶于吡啶中，然后加入DDTT(976.77mg,4.76mmol)，加毕反应体系搅拌2h。所得棕色反应液用乙酸乙酯(200mL)稀释，依次用饱和碳酸氢钠溶液(50x3mL)和饱和食盐水洗涤，分离有机相，无水硫酸钠干燥，过滤，滤液浓缩，得粗品，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～1/4)，得化合物4-1。
MS(ESI)m/z(M/2+H)
+＝742.1
步骤2：化合物4-2的制备
化合物4-1(2g,1.21mmol)加入到80％的乙酸(36mL)和乙腈(10mL)混合溶液中，40℃条件下搅拌反应20h，反应液用乙酸乙酯(300mL)稀释，小心地加入饱和碳酸氢钠溶液调节pH至9.0，分离有机相，用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，减压浓缩，粗品用乙酸乙酯(10mL)打浆，过滤，滤饼用乙酸乙酯(2x 2mL)洗涤，真空干燥得化合物4-2。
MS(ESI)m/z(M+H)
+＝878.3.
1H NMR(400MHz,DMSO-d
6)δ8.74-8.25(m,4H),8.08-8.02(m,5H),7.57-7.48(m,7H),6.44-6.37(m,2H),5.82-5.69(m,1H),5.56-5.51(m,2H),5.05-4.95(m,1H),4.55-4.43(m,2H),4.35-4.25(m,4H),3.84- 3.64(m,2H),2.88-2.84(m,2H).
31P NMR(162MHz,DMSO-d
6)δ69.59-67.63
19F NMR(376MHz,DMSO-d
6)δ-203--207
步骤3：化合物4-3的制备
化合物4-2(400mg,455.70μmol)溶于乙腈(2mL)中，依次加入4A分子筛(0.3g)和四氮唑(0.45M的乙腈溶液,10.13mL)，所得反应混合物用氩气鼓泡4min，然后滴加入2-氰乙基N,N,N',N'-四异丙基亚磷酰二胺(206.03mg,683.55μmol)，反应体系搅拌反应1h。乙酸乙酯(60mL)稀释，过滤，有机相依次用饱和碳酸氢钠(20mL x 3)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，得粗品4-3。
MS(ESI)m/z(M+H)
+＝977.2.
步骤4：化合物4-4的制备
化合物4-3(0.43g,440.21μmol)溶于二氯甲烷(30mL)中，所得混合物用氩气鼓泡4min，15℃搅拌10min后冷至0℃，滴加入硼烷二甲硫醚络合物(2M的四氢呋喃溶液,660.32μL)加毕反应体系升温至15℃搅拌30min。加水(20mL)淬灭反应，加二氯甲烷(50mL)稀释，室温搅拌30min后分液，有机相用饱和食盐水(10mL)洗涤，无水硫酸镁干燥，过滤，滤液减压浓缩，得粗品4-4，不经进一步纯化直接用于下一步反应。
步骤5：化合物4-5的制备
将化合物4-4(420mg,423.97μmol)溶于乙醇(3mL)和乙腈(3mL)的混合溶液中，加入叔丁胺(6mL)，反应室温搅拌3h。反应体系减压浓缩，得粗品4-5，不经进一步纯化直接用于下一步反应。
步骤6：化合物4A，4B，4C和4D的制备
将化合物4-5(350mg,395.70μmol)溶于30％的甲胺乙醇溶液(40mL)中，反应室温搅拌反应24h。反应体系减压浓缩，残渣溶于水(10mL)中，乙酸乙酯(5mL x 3)反萃，水相减压浓缩后经高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*30mm 10μm；流动相:[水(10mM碳酸氢铵)-乙腈]；乙腈％:0％-30％,流速：25mL/min，20min)。得：
化合物4A(HPLC保留时间5.4min)
化合物4B(HPLC保留时间5.9min)
化合物4C(HPLC保留时间6.7min)
化合物4D(HPLC保留时间7.4min)
化合物4A：
MS(ESI)m/z(M+H)
+＝677.2.
1H NMR(400MHz,D
2O)δ8.36(s,1H),8.27(s,1H),7.96(s,1H),7.91(s,1H),6.29-6.23(m,2H),5.80(d,J＝51.2Hz,1H),5.45(d,J＝51.2Hz,1H),4.85-4.72(m,2H),4.38-4.32(m,2H),4.24-4.15(m,1H),3.91-3.87(m,2H),0.50--0.20(br,3H).
31P NMR(162MHz,D
2O)δ96.2-91.9,54.5.
19F NMR(376MHz,D
2O)δ-202.7--203.0.
化合物4B：
MS(ESI)m/z(M+H)
+＝677.2.
1H NMR(400MHz,D
2O)δ8.01(s,1H),7.98(s,1H),7.86(br s,1H),7.46(br s,1H),6.37(d,J＝13.6Hz,1H),6.16(d,J＝14.0Hz,1H),5.38-5.07(m,2H),4.42-4.32(m,2H),4.32-4.25(m,1H),4.25-4.18(m,1H),3.87-3.79(m,2H),0.25--0.30(br,3H).
31P NMR(162MHz,D
2O)δ94.7-91.5,54.062.
19F NMR(376MHz,D
2O)δ-202.5--204.1.
化合物4C：
MS(ESI)m/z(M+H)
+＝677.2.
1H NMR(400MHz,D
2O)δ8.20(s,1H),8.07(s,1H),7.92(s,1H),7.75(s,1H),6.16-6.08(m,2H),5.81(d,J＝50.8Hz,1H),5.40(d,J＝51.2Hz,1H),5.03-4.81(m,2H),4.38-4.28(m,4H),3.96-3.88(m,2H),0.50--0.20(br,3H).
31P NMR(162MHz,D
2O)δ94.3-91.1,54.033.
19F NMR(376MHz,D
2O)δ-201.377--202.447.
化合物4D：
MS(ESI)m/z(M+H)
+＝677.1.
1H NMR(400MHz,D
2O)δ8.01(s,1H),7.92(s,2H),7.77(s,2H),6.19-6.16(m,2H),5.31-5.18(m,2H),4.81-4.72(m,2H),4.38-4.27(m,4H),3.85-3.77(m,2H),0.50--0.10(br,3H).
31P NMR(162MHz,D
2O)δ94.8-91.5,54.011.
19F NMR(376MHz,D
2O)δ-202.724--202.889.
实施例5：化合物5A、5B、5C、5D的制备
步骤1：化合物5-2的制备
0℃条件下，化合物5-1(20g,76.84mmol)溶于乙腈(300mL)中，加入氢化钠(4.61g,115.26mmol,60％)，搅拌0.5h后，加入苄基溴(13.14g,76.84mmol)，升温至20℃搅拌反应3h。加入甲醇淬灭反应，同时加入水(100mL)和乙酸乙酯(150mL)，分出有机相，经无水硫酸钠干燥，滤除固体，滤液浓缩，粗品用石油醚打浆，分离出固体得化合物5-2.
1H NMR(400MHz,CDCl
3)δ7.41-7.30(m,5H),5.74(d,J＝3.8Hz,1H),4.76(d,J＝11.8Hz,1H),4.60-4.55(m,2H),4.35(dt,J＝3.2,7.0Hz,1H),4.13(dd,J＝3.2,8.8Hz,1H),4.02-3.92(m,2H),3.87(dd,J＝4.6,8.8Hz,1H),1.58(s,3H),1.37(s,3H),1.35(d,J＝4.4Hz,6H)
步骤2：化合物5-3的制备
化合物5-2(51g,145.55mmol)溶于水(42mL)和乙酸(179.55g,2.99mol,171mL)中，反应于20℃搅拌反应72h。反应液用1.0M的氢氧化钠溶液中和，乙酸乙酯(300mL x 3)萃取，合并有机相，经无水硫酸钠干燥，滤除固体，滤液浓缩得粗品5-3，不经进一步纯化直接用于下一步反应。
1H NMR(400MHz,CDCl
3)δ7.40-7.32(m,5H),5.74(d,J＝3.8Hz,1H),4.77(d,J＝11.2Hz,1H),4.59(t,J＝4.0Hz,1H),4.54(d,J＝11.2Hz,1H),4.14-4.06(m,2H),3.91(dd,J＝8.8,4.4Hz,1H),3.72-3.61(m,2H),2.56(br s,2H),1.57(s,3H),1.34(s,3H).
步骤3：化合物5-4的制备
化合物5-3(20g,64.45mmol)的水(200mL)溶液加入到高碘酸钠(15.99g,74.76mmol)的水(100mL)溶液中，反应于0℃搅拌反应1h。然后加入乙二醇(2.60g,41.89mmol,2.34mL)继续搅拌20min，反应液用乙酸乙酯(170mL x 3)萃取，合并有机相，经无水硫酸钠干燥，滤除固体，滤液浓缩得粗品5-4，不经进一步纯化直接用于下一步反应。
1H NMR(400MHz,CDCl
3)δ9.60(d,J＝1.8Hz,1H),7.35-7.32(m,5H),5.80(d,J＝3.6Hz,1H),4.76-4.70(m,1H),4.65-4.60(m,1H),4.58(t,J＝3.8Hz,1H),4.47(dd,J＝9.0,1.6Hz,1H),3.83(dd,J＝9.4,4.4Hz,1H),1.59(s,3H),1.36(s,3H).
步骤4：化合物5-5的制备
0℃条件下，化合物5-4(17.94g,64.46mmol)溶于二氧六环(45mL)和水(40mL)中，加入福尔马林(36mL,483.54mmol,37％的水溶液)和氢氧化钠(1M的水溶液,176mL)，反应升温到20℃搅拌反应48h。反应液用乙酸乙酯(200mL x 3)萃取，合并有机相，经无水硫酸钠干燥，滤除固体，滤液浓缩得粗品5-5，不经进一步纯化直接用于下一步反应。
1H NMR(400MHz,CDCl
3)δ7.38-7.31(m,5H),5.78-5.72(m,1H),4.83-4.76(m,1H),4.66-4.61(m,1H),4.55(d,J＝11.7Hz,1H),4.22-4.16(m,1H),3.95-3.85(m,2H),3.80-3.74(m,1H),3.62-3.50(m,1H),2.37(t,J＝6.9Hz,1H),1.88(dd,J＝3.7,9.6Hz,1H),1.62(s,3H),1.32(s,3H).
步骤5：化合物5-6的制备
化合物5-5(19g,61.22mmol)溶于二异丙醚(350mL)中，加入脂肪酶Novozyme-435(1.5g,61.22mmol)和乙酸乙烯酯(5.27g,61.22mmol,5.67mL)，反应升温到50℃搅拌反应16h。反应液用乙酸乙酯(200mL x 3)萃取，合并有机相，经无水硫酸钠干燥，滤除固体，滤液浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～0/1)，得化合物5-6.
1H NMR(400MHz,CDCl
3)δ7.34(m,5H),5.75(d,J＝4.0Hz,1H),4.80(d,J＝11.8Hz,1H),4.65(t,J＝4.6Hz,1H),4.51(d,J＝11.8Hz,1H),4.25(d,J＝11.8Hz,1H),4.12-4.06(m,1H),4.00(d,J＝5.4Hz,1H),3.93(br d,J＝6.8Hz,2H),2.36(t,J＝6.8Hz,1H),2.01-1.97(m,3H),1.62(s,3H),1.33(s,3H).
步骤6：化合物5-7的制备
化合物5-6(5g,14.19mmol)溶于吡啶(21mL)和二氯甲烷(83mL)中，加入对甲苯磺酰氯(2.98g,15.61mmol)，反应于15℃搅拌反应24h。反应液用10％的盐酸溶液淬灭，二氯甲烷(40mL x 3)萃取，合并有 机相，经无水硫酸钠干燥，滤除固体，滤液浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～1/1)，得化合物5-7.
1H NMR(400MHz,CDCl
3)δ7.79(br d,J＝7.9Hz,2H),7.37-7.30(m,7H),5.68(d,J＝3.6Hz,1H),4.69(br d,J＝12.1Hz,1H),4.56(br t,J＝4.2Hz,1H),4.54-4.45(m,2H),4.33(d,J＝10.6Hz,1H),4.16(d,J＝11.8Hz,1H),4.12-4.08(m,1H),4.00-3.93(m,2H),2.41(s,3H),1.89(s,3H),1.34(s,3H),1.26(s,3H).
步骤7：化合物5-8的制备
0℃条件下，化合物5-7(20g,40.61mmol)溶于乙酸(200mL)中，加入乙酸酐(38.03mL,406.06mmol)和硫酸(216.44μL,4.06mmol)，反应升温至20℃搅拌反应6h。反应液倾入水(600mL)中，用氢氧化钠溶液中和至pH＝7，乙酸乙酯(200mL x 3)萃取，合并有机相，经饱和食盐水(200mL)洗涤，无水硫酸钠干燥，滤除固体，滤液浓缩得粗品5-8，不经进一步纯化直接用于下一步反应。
步骤8：化合物5-9的制备
化合物N-(5H-嘌呤-6-基)苯甲酰胺(5.87g,24.52mmol)溶于二氯乙烷(200mL)中，加入N,O-双三甲硅基乙酰胺(16.16mL,65.39mmol)，升温至80℃搅拌0.5h，冷至0℃，然后加入化合物5-8(9.0g,16.35mmol)和三氟甲磺酸三甲基硅酯(4.73mL,26.15mmol)，升温至80℃搅拌反应1h。反应液冷至室温，倾入饱和碳酸氢钠溶液(200mL)中，乙酸乙酯(200mL x 2)萃取，有机相用饱和食盐水洗涤(200mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～0/1)，得化合物5-9.
MS(ESI)m/z(M+H)
+＝730.3.
步骤9：化合物5-10的制备
化合物5-9(11g,10.85mmol)溶于水(24mL)和二氧六环(24mL)中，加入氨水(33.44mL,217.06mmol,25-28％)和2.0M的氢氧化钠水溶液(31.68mL)，反应于25℃搅拌18h。反应液倾入水(100mL)中，乙酸乙酯(40mL x 3)萃取，有机相用饱和食盐水洗涤(50mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～10/1)，得化合物5-10.
MS(ESI)m/z(M+H)
+＝474.1.
步骤10：化合物5-11的制备
化合物5-10(1.68g,3.38mmol)溶于氨水(19.31mL)中，反应于30℃搅拌24h。反应液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～10/1)，得化合物5-11.
MS(ESI)m/z(M+H)
+＝370.2
步骤11：化合物5-12的制备
化合物5-11(1.4g,3.79mmol)溶于甲醇(140mL)中，加入氢氧化钯/碳(0.45g,640.84μmol,20％湿)，反应于60℃氢气氛条件下搅拌2h，然后加入甲酸铵(1.91g,30.32mmol)，继续搅拌反应15h。反应液过滤除去催化剂，滤液减压浓缩得粗品5-12。不经进一步纯化直接用于下一步反应。
1H NMR(400MHz,DMSO-d
6)δ8.22(s,1H),8.14(s,1H),7.32(s,2H),5.89(s,1H),4.55(s,1H),4.40(s,1H),4.25(s,1H),3.93–3.91(m,1H),3.81-3.74(m,4H)
步骤12：化合物5-13的制备
0℃，氮气保护条件下，化合物5-12(1g,3.58mmol)溶于吡啶(20mL)和N,N-二甲基甲酰胺(10mL)中，加入三甲基氯硅烷(1.97g,18.12mmol,2.3mL)，搅拌30min后，加入苯甲酰氯(968.00mg,6.89mmol,0.8mL)，反应于20℃搅拌3h。反应液用水(10mL)和氨水(10mL)淬灭，搅拌30min后用乙酸乙酯(20mL x 3)萃取，有机相用无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(乙酸乙酯/甲醇(v/v)＝20/3)，得化合物5-13.
MS(ESI)m/z(M+H)
+＝384.0
1H NMR(400MHz,CDCl
3)δ9.24(br s,1H),8.71(s,1H),8.41(s,1H),7.99(br d,J＝7.5Hz,2H),7.65-7.59(m,1H),7.52(t,J＝7.5Hz,2H),6.12(s,1H),4.68-4.62(m,2H),4.09-4.05(m,1H),3.99(s,2H),3.89(d,J＝8.0Hz,1H).
步骤13：化合物5-14的制备
20℃，氩气保护条件下，化合物5-13(500mg,1.30mmol)溶于吡啶(10mL)中，加入DMTrCl(530mg,1.56mmol)，搅拌16h，反应液用甲醇(10mL)淬灭，减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝20/1～10/1)，得化合物5-14.
MS(ESI)m/z(M+H)
+＝686.2
步骤14：化合物5-15的制备
20℃，氩气保护条件下，化合物5-14(500mg,729.16μmol)、四氮唑(0.45M的乙腈溶液,25.00mL)和
分子筛分散于乙腈(4mL)中，加入化合物1-6(836.68mg,955.20μmol)的乙腈(2mL)溶液，反应搅拌1h，反应液滤除分子筛，加乙酸乙酯(50mL)稀释，有机相依次用饱和碳酸氢钠水溶液(50mL x 2)，饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝100/3)，得化合物5-15.
MS(ESI)m/z(M/2+H)
+＝731.0.
步骤15：化合物5-16的制备
0℃，氩气保护条件下，化合物5-15(1.3g,890.13μmol)、
分子筛(300mg)分散于二氯甲烷(30mL)中，滴加入硼烷二甲硫醚(2M的四氢呋喃溶液,1.34mL)，反应于0℃搅拌15min。反应液加二氯甲烷(50mL)稀释，滤除分子筛，滤液依次用水(50mL)、饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/乙酸乙酯(v/v)＝1/1)，得化合物5-16.
步骤16：化合物5-17的制备
化合物5-16(850mg,576.55μmol)溶于二氯甲烷(10mL)中，加入2,2-二氯乙酸的二氯甲烷(10.53g,2.31mmol,2mL,5％)，搅拌反应0.5h，加入二氯甲烷(50mL)稀释，有机相用饱和碳酸氢钠溶液(20mL x 2)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经薄层色谱(二氯甲烷/甲醇(v/v)＝10/1)分离纯化，得化合物5-17。
MS(ESI)m/z(M-14+H)
+＝856.4.
步骤17：化合物5-18的制备
氩气保护，20℃条件下，化合物5-17(400mg,467.44μmol)、
分子筛(1g)和四氮唑(0.45M的乙腈溶液,16mL)分散于乙腈(2mL)和四氢呋喃(3mL)中，滴加入2-氰乙基N,N,N',N'-四异丙基亚磷酰二胺(189.80mg,629.71μmol,0.2mL)的乙腈(0.5mL)溶液，搅拌反应1h。反应液滤除分子筛，滤液用乙酸乙酯(20mL)稀释，有机相依次用饱和碳酸氢钠水溶液(20mL x 2)和饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经薄层色谱(二氯甲烷/甲醇(v/v)＝15/1)分离纯化，得化合物5-18。
步骤18：化合物5-19的制备
0℃，氩气保护条件下，化合物5-18(230mg,237.46μmol)、
分子筛(100mg)分散于四氢呋喃(3mL)和二氯甲烷(2mL)中，滴加入硼烷二甲硫醚(2M的四氢呋喃溶液,460.00μL)，反应升至15℃搅拌10min。反应液用二氯甲烷(20mL)稀释，过滤，滤液用水(30mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩得粗品5-19，不经进一步纯化直接用于下一步反应。
步骤19：化合物5A,5B,5C和5D的制备
化合物5-19(200mg,203.58μmol)溶于甲胺的水溶液(10mL,33％)中，20℃搅拌24h，反应液用乙酸乙酯(30mL)萃取，水相冻干，粗品经高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*40mm 10μm；流动相:[水(10mM碳酸氢铵)-乙腈]；乙腈％:0％-20％,流速：25mL/min，25min)。得：
化合物5A(HPLC保留时间5.59min)
化合物5B(HPLC保留时间5.87min)
化合物5C(HPLC保留时间6.16min)
化合物5D(HPLC保留时间7.44min)
化合物5A：
MS(ESI)m/z(M-H)
-＝666.8
1H NMR(400MHz,D
2O)δ8.11(s,2H),8.02(s,1H),7.99(s,1H),6.27(d,J＝16.1Hz,1H),6.01(s,1H),5.58-5.42(m,1H),4.84-4.72(m,1H),4.67(br s,1H),4.46(br d,J＝10.5Hz,1H),4.35(br d,J＝9.0Hz,1H),4.20-4.10(m,2H),4.07-3.88(m,4H),0.52--0.57(m,6H).
19F NMR(376MHz,D
2O)-202.51--202.65.
31P NMR(162MHz,D
2O)δ94.12-90.98.
化合物5B：
MS(ESI)m/z(M-H)
-＝667.1
1H NMR(400MHz,D
2O)δ8.26(s,1H),8.15-8.09(m,1H),8.07(s,1H),7.96(s,1H),6.30(d,J＝16.4Hz,1H),6.05(s,1H),5.48-5.42(m,0.5H),5.33(br s,0.5H),5.02-4.86(m,1H),4.75(s,1H),4.51(br d,J＝10.5Hz,1H),4.33(br d,J＝9.0Hz,1H),4.23(br d,J＝12.0Hz,1H),4.15(br d,J＝12.2Hz,1H),4.04(br d,J＝5.4Hz,1H),3.95-4.02(m,2H),3.91(d,J＝8.1Hz,1H),0.46--0.33(m,6H).
19F NMR(376MHz,D
2O)-201.02--201.20.
31P NMR(162MHz,D
2O)δ94.86-93.72.
化合物5C：
MS(ESI)m/z(M-H)
-＝666.8.
1H NMR(400MHz,D
2O)δ8.40(s,1H),7.89(s,1H),7.82(br s,1H),7.75(br s,1H),6.29(br d,J＝13.7Hz,1H),6.02(s,1H),5.59-5.23(m,1H),4.73(br s,1H),4.71-4.68(m,2H),4.39-4.28(m,2H),4.21-4.08(m,1H),4.07-3.90(m,3H),3.88(br d,J＝7.3Hz,1H),0.18(br s,6H).
19F NMR(376MHz,D
2O)-204.17--204.48.
31P NMR(162MHz,D
2O)δ96.82-89.33.
化合物5D：
MS(ESI)m/z(M-H)
-＝666.8.
1H NMR(400MHz,D
2O)δ8.45(s,1H),8.32(s,1H),8.07(br s,1H),8.05(br s,1H),6.46(d,J＝14.7Hz,1H),6.17(s,1H),5.64-5.39(m,1H),5.18-5.05(m,1H),4.98(br d,J＝8.8Hz,1H),4.90(s,1H),4.53-4.36(m,3H),4.19-4.14(m,2H),4.10(br dd,J＝5.6,12.0Hz,1H),4.02(br d,J＝8.1Hz,1H),0.61-0.11(m,6H).
19F NMR(376MHz,D
2O)-201.73--202.52.
31P NMR(162MHz,D
2O)δ95.28–93.68.
实施例6：化合物6A、6B、6C、6D的制备
步骤1：化合物6-1的制备
20℃条件下，化合物D-阿拉伯糖(25g,166.5mmol)溶于DMF(250mL)中，加入咪唑(17g,249.8mmol)和叔丁基二苯基氯硅烷(45.8g,166.5mmol)，搅拌2h。溶液倾入水(2.5L)中，加乙酸乙酯(1000mL)萃取，无水硫酸钠干燥，过滤，滤液减压浓缩得粗品6-1，不经进一步纯化直接用于下一步反应。
MS(ESI)m/z(M+Na)
+＝411.2.
步骤2：化合物6-2的制备
化合物6-1(17.5g,45.0mmol)溶于无水丙酮(150mL,2.04mol)中，依次加入无水硫酸铜(20g,125.3mmol)和硫酸(0.8mL,98％)，反应混合物于20℃搅拌17h。过滤，滤液用氢氧化钙中和，再次过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～7/3)，得化合物6-2.
MS(ESI)m/z(M+Na)
+＝451.2.
1H NMR(400MHz,CDCl
3)7.70-7.65(m,4H),7.45-7.37(m,6H),5.89(d,J＝4.2Hz,1H),4.56(d,J＝4.2Hz,1H),4.45-4.44(m,1H),4.09-4.04(m,1H),3.85-3.81(m,2H),1.33(s,3H),1.30(s,3H),1.07(s,9H).
步骤3：化合物6-3的制备
氮气保护下，化合物6-2(27g,63.0mmol)溶于四氢呋喃(300mL)中，依次加入溴化苄(44mL,370.8mmol)和氢氧化钾(31.8g,565.9mmol)，反应混合物加热至70℃搅拌17h。冷至室温，过滤，滤渣用四氢呋喃(20mL x 3)洗涤，合并滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～9/1)，得化合物6-3.
MS(ESI)m/z(M+Na)
+＝393.1.
1H NMR(400MHz,CDCl
3)7.38-7.28(m,10H),5.92(d,J＝3.9Hz,1H),4.66(d,J＝4.2Hz,1H),4.62-4.55(m,4H),4.29-4.28(m,1H),4.04(d,J＝2.9Hz,1H),3.65(d,J＝6.1Hz,2H),1.45(s,3H),1.33(s,3H).
步骤4：化合物6-4的制备
化合物6-3(33.5g,90.4mmol)溶于甲醇(250mL)中，加入D-樟脑磺酸(0.1g,399.5μmol)，反应混合物加热至70℃搅拌12h。冷至室温，滴加入30滴三乙胺，反应液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～7/3)，得化合物6-4.
MS(ESI)m/z(M+Na)
+＝367.0.
1H NMR(400MHz,CDCl
3)7.39-7.23(m,10H),4.92(s,0.6H),4.87(d,J＝4.6Hz,0.4H),4.79-4.44(m,4H),4.28(q,J＝2.2Hz,1H),4.16-4.09(m,1H),3.89-3.82(m,1H),3.66-3.65(m,0.6H),3.54(d,J＝5.6Hz,0.7H),3.45-3.44(m,0.4H),3.44-3.41(s,3H),3.37-3.36(m,0.6H),2.61-2.59(m,0.3H).
步骤5：化合物6-5的制备
0℃条件下，化合物6-4(1.24g,3.6mmol)溶于二氯甲烷(40mL)中，依次加入吡啶(2.7mL 33.5mmol)和三氟甲磺酸酐(0.8mL,4.9mmol)，20min后，加水(10mL)淬灭反应，分液，有机相用无水硫酸钠干燥，减压浓缩得粗品，粗品溶于四氢呋喃(25mL)中，冷至0℃，体系中加入四丁基氟化铵(1M的四氢呋喃溶液,18mL)，升温至20℃搅拌12h，反应液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～9/1)，得化合物6-5.
MS(ESI)m/z(M+Na)
+＝369.2.
1H NMR(400MHz,CDCl
3)7.40-7.28(m,10H),5.05-4.99(m,1H),4.74-4.47(m,5H),4.32(m,1H),4.16-4.04(m,1H),3.69-3.63(m,1H),3.58-3.51(m,1H),3.34(s,3H).
19F NMR(376MHz,CDCl
3)-209.4.
步骤6：化合物6-6的制备
20℃条件下，化合物6-5(8.4g,24.2mmol)溶于三氟乙酸(90mL,1.2mol)和水(10mL,555.1mmol)中，搅拌12h后，加水(150mL)和二氯甲烷(200mL)于溶液中，水相用10M的氢氧化钠调节至pH＝7.0，分出 有机相，水相用二氯甲烷(50mL x 2)萃取，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩得粗品，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～7/3)，得化合物6-6.
MS(ESI)m/z(M+Na)
+＝355.0.
19F NMR(376MHz,CDCl
3)-206.8.
步骤7：化合物6-7的制备
-50℃，氩气保护条件下，化合物6-6(5.9g,17.8mmol)和四氯化碳(7.7mL 80.3mmol)溶于甲苯(70mL)中，滴加入三(二甲胺基)膦(3.49g,21.41mmol,3.89mL)的甲苯(5mL)溶液，反应升温至0℃搅拌3h，然后冷至-20℃，加入冷的甲苯(30mL)稀释，滴加入冷的饱和食盐水(30mL)淬灭反应，分出有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩得粗品6-7，粗品不经进一步纯化直接用于下一步反应.
MS(ESI)m/z(M+NH
4)
+＝369.1.
1H NMR(400MHz,CDCl
3)7.27-7.26(m,10H),6.30-6.29(m,1H),5.16-4.95(m,1H),4.89-4.85(m,1H),4.72-4.43(m,4H),4.12–4.11(m,1H),3.80-3.50(m,2H).
19F NMR(376MHz,CDCl
3)-200.5--200.6.
步骤8：化合物6-8的制备
20℃，氩气保护条件下，化合物6-7(6.5g,18.5mmol)、4-氯-5-氟-7H-吡咯并[2,3-d]嘧啶(3.2g,18.5mmol)溶于乙腈(100mL)中，依次加入氢氧化钾(3.1g,55.6mmol)和三(3,6-二氧杂庚基)胺(599.3mg,1.9mmol)，搅拌反应12h后，反应液浓缩，粗品溶于乙酸乙酯(150mL)和水(60mL)中，分液，水相用乙酸乙酯(20mL x 2)萃取，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(石油醚/乙酸乙酯(v/v)＝1/0～4/1)，得化合物6-8.
MS(ESI)m/z(M+H)
+＝486.1.
1H NMR(400MHz,CDCl
3)8.61(s,1H),7.41-7.30(m,11H),6.67-6.60(m,1H),4.81-4.74(m,1H),4.65-4.50(m,4H),4.43-4.33(m,2H),3.92-3.84(m,1H),3.68-3.65(m,1H).
19F NMR(376MHz,CDCl
3)-165.8--165.9,-203.8--204.5.
步骤9：化合物6-9的制备
-70℃，氮气保护条件下，化合物6-8(0.1g,179.1μmol)溶于二氯甲烷(5mL)中，滴加入三氯化硼(1M的正庚烷溶液,0.9mL)，该温度下搅拌反应1h。缓慢加入甲醇(15mL)淬灭反应，升至室温，减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～20/1)，得化合物6-9.
1H NMR(400MHz,DMSO-d
6)8.73(s,1H),8.03(d,J＝1.5Hz,1H),6.52(d,J＝15.4Hz,1H),5.76(d,J＝6.1Hz,1H),5.35-5.16(m,2H),4.44-4.30(m,1H),4.01-3.93(m,1H),3.80-3.55(m,2H).
19F NMR(376MHz,DMSO-d
6)-169.15,-204.15--204.64.
步骤10：化合物6-10的制备
10℃条件下，化合物6-9(0.33g,1.1mmol)溶于甲醇钠的甲醇溶液(0.5M,16.5mL)中，该温度下搅拌反应12h。减压浓缩除去溶剂，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～20/1)，得化合物6-10.
MS(ESI)m/z(M+H)
+＝302.0.
1H NMR(400MHz,DMSO-d
6)8.48(s,1H),7.67(d,J＝2.0Hz,1H),6.50-6.43(m,1H),5.73(d,J＝6.0Hz,1H),5.32-5.28(m,0.5H),5.17(t,J＝5.3Hz,1.5H),4.41-4.30(m,1H),4.10-4.04(m,3H),3.95-3.94(m,1H),3.72-3.70(m,1H),3.60-3.55(m,1H).
19F NMR(376MHz,DMSO-d
6)-166.9,-204.8.
步骤11：化合物6-11的制备
20℃，氩气保护条件下，化合物6-10(0.22g,730.3μmol)溶于乙腈(20mL)中，依次加入碘化钠(547.3mg,3.6mmol)和三甲基氯硅烷(463μL 3.6mmol)，该温度下搅拌反应3h。冷至0℃，加入甲醇(3mL)淬灭反应，搅拌反应5min后，减压浓缩除去溶剂，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～10/1)，得化合物6-11.
MS(ESI)m/z(M+H)
+＝288.0.
1H NMR(400MHz,DMSO-d
6)12.19(s,1H),7.96(s,1H),7.35(s,1H),6.34-6.31(m,1H),5.70(br s,1H),5.25-5.18(m,1H),5.11-5.05(m,1H),4.34-4.28(m,1H),3.92(s,1H),3.75-3.53(m,2H).
19F NMR(376MHz,DMSO-d
6)-165.3,-204.8--205.0.
步骤12：化合物6-12的制备
20℃，氩气保护条件下，化合物6-11(0.36g,1.25mmol)溶于吡啶(8mL)中，加入DMTrCl(0.26g,767.4μmol)，该温度下搅拌反应4h。加入甲醇(3mL)淬灭反应，搅拌5min后，减压浓缩除去溶剂，粗品溶于乙酸乙酯(60mL)中，有机相用饱和食盐水(15mL x 3)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～20/1)，得化合物6-12.
MS(ESI)m/z(M+H)
+＝592.2.
1H NMR(400MHz,DMSO-d
6)12.23(s,1H),7.98(s,1H),7.39-7.18(m,10H),6.86-6.83(m,4H),6.37–6.32(m,1H),5.70(d,J＝6.8Hz,1H),5.36-5.17(m,1H),4.52-4.40(m,1H),4.09-4.02(m,1H),3.73(s,6H),3.29-3.19(m,2H).
19F NMR(376MHz,DMSO-d
6)-165.4,-201.9.
步骤13：化合物6-13的制备
20℃，氩气保护条件下，化合物6-12(0.25g,424.0μmol)溶于乙腈(5mL)中，依次加入四氮唑(0.45M的乙腈溶液,10mL)和
分子筛(0.6g)，搅拌10min后，加入1-6(0.65g,742.1μmol)的乙腈溶液，继续搅拌反应50min后，混合物倾入乙酸乙酯(30mL)中，过滤，滤液依次用饱和碳酸氢钠水溶液(10mL)，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～20/1)，得化合物6-13.
步骤14：化合物6-14的制备
氩气保护，25℃条件下，化合物6-13(0.68g,498.4μmol)溶于二氯甲烷(5mL)中，加入
分子筛(0.6g)，搅拌30min后，体系滴加入硼烷二甲硫醚络合物(2M的四氢呋喃溶液,747.63μL)，加毕反应体系25℃条件下搅拌20min。反应液用二氯甲烷(40mL)稀释，过滤，滤液依次用水(10mL)、饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，得粗品6-14，不经进一步纯化直接用于下一步反应。
步骤15：化合物6-15的制备
化合物6-14(0.73g,529.7μmol)溶于二氯甲烷(5mL)中，加入2，2-二氯乙酸(5mL,2.6mmol,5％的二氯甲烷溶液)，20℃条件下搅拌反应30min，体系中加入三乙基硅烷(5mL,31.3mmol)，继续搅拌反应30min。反应液倾入二氯甲烷(60mL)，然后依次用饱和碳酸氢钠溶液(10mL x 2)、饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经硅胶柱层析纯化(二氯甲烷/甲醇(v/v)＝1/0～20/1)，得化合物6-15。
MS(ESI)m/z(M+H)
+＝774.3.
1H NMR(400MHz,DMSO-d
6)12.26(s,1H),11.25(s,1H),8.76(s,1H),8.58-8.57(m,1H),8.03–8.02(m,2H),7.95–7.94(m,1H),7.69-7.62(m,1H),7.59-7.51(m,2H),7.35-7.34(m,1H),6.49-6.31(m,2H),6.01(d,J＝6.5Hz,1H),5.67-5.66(m,1H),5.61-5.52(m,1H),5.45–5.44(m,1H),5.36–5.35(m,1H),5.09–5.08(m,1H),4.85-4.69(m,1H),4.51-4.40(m,1H),4.39-4.31(m,1H),4.27-4.10(m,4H),3.56–3.55(m,2H),2.89-2.88(m,2H)..
19F NMR(376MHz,DMSO-d
6)-164.8,-201.3,-206.9.
31P NMR(162MHz,DMSO-d
6)δ115.0-115.1.
步骤16：化合物6-16的制备
氩气保护，20℃条件下，化合物6-15(0.22g,284.5μmol)、
分子筛(1g)和四氮唑(0.45M的乙腈溶液,10mL,4.5mmol)分散在乙腈(2mL)和四氢呋喃(5mL)的混合溶液中，滴加入2-氰乙基N,N,N',N'-四异丙基亚磷酰二胺(120μL,377.8μmol)的乙腈(0.5mL)溶液，反应体系搅拌反应1h。反应液用乙酸乙酯(50mL)稀释，过滤，滤液用水(20mL x 3)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品经制备薄层层析色谱(二氯甲烷/甲醇(v/v)＝10/1)分离纯化，得化合物6-16。
步骤17：化合物6-17的制备
氩气保护，0℃条件下，硼烷二甲硫醚络合物(2M的四氢呋喃溶液,0.2mL,0.4mmol)滴加入化合物6-16(0.1g,114.6μmol)和
分子筛(0.5g)的四氢呋喃(5mL)溶液中，加毕反应体系于15℃条件下搅拌30min。加乙酸乙酯(40mL)稀释，过滤，滤液用水(15mL x 3)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩得粗品6-17，不经进一步纯化直接用于下一步反应。
MS(ESI)m/z(M+H)
+＝887.2
步骤18：化合物6A，6B，6C和6D的制备
将化合物6-17(0.11g,124.1μmol)溶于33％的甲胺乙醇溶液(3mL)中，反应15℃条件下搅拌反应18h。反应体系减压浓缩，残渣溶于水(10mL)中，用乙酸乙酯(30mL)反萃，水相冻干，所得粗品经高效制备液相分离(分离条件:色谱柱:Xbridge Prep OBD C18 150*40mm 10μm；流动相:[水(0.05％氢氧化铵)-乙腈]；乙腈％:0％-30％,流速：25mL/min，20min)。得：
化合物6A(HPLC保留时间6.05min)
化合物6B(HPLC保留时间6.47min)
化合物6C(HPLC保留时间6.39min)
化合物6D(HPLC保留时间7.47min)
化合物6A：
MS(ESI)m/z(M-H)
-＝674.7
1H NMR(400MHz,D
2O)δ8.43(s,1H),8.12(s,1H),7.93(s,1H),7.24(s,1H),6.49-6.38(m,2H),5.87-5.70(m,1H),5.38-5.19(m,1H),5.19-5.03(m,1H),4.86-4.81(m,1H),4.55-4.37(m,2H),4.32-4.18(m,2H), 4.07-3.94(m,2H),0.62--0.39(m,6H).
19F NMR(376MHz,D
2O)-165.05,-201.16--201.51,-203.00--203.35.
31P NMR(162MHz,D
2O)δ95.50-90.45.
化合物6B：
MS(ESI)m/z(M-H)
-＝674.8
1H NMR(400MHz,D
2O)δ8.50(s,1H),8.18(s,1H),7.95(s,1H),7.20(s,1H),6.56-6.37(m,2H),5.63-5.40(m,1H),5.39-5.14(m,2H),4.53-4.35(m,3H),4.35-4.23(m,2H),4.10-3.91(m,2H),0.28(br s,6H).
19F NMR(376MHz,D
2O)-164.19,-199.92--200.68,-201.18--202.10.
31P NMR(162MHz,D
2O)δ95.93-91.70.
化合物6C:
MS(ESI)m/z(M-H)
-＝674.7
1H NMR(400MHz,D
2O)δ8.39(s,1H),8.15(s,1H),7.93-7.83(m,1H),7.20(s,1H),6.53-6.35(m,2H),5.71-5.49(m,1H),5.37-5.16(m,1H),5.13-4.90(m,2H),4.55-4.36(m,3H),4.31-4.18(m,1H),4.10-3.96(m,2H),0.36(br s.,6H).
19F NMR(376MHz,D
2O)-165.35,-199.73--200.19,-202.31--202.91.
31P NMR(162MHz,D
2O)δ96.82-90.33.
化合物6D:
MS(ESI)m/z(M-H)
-＝674.7
1H NMR(400MHz,D
2O)δ8.25(s,1H),8.06(s,1H),7.66(s,1H),7.05(s,1H),6.29-6.15(m,2H),5.75-5.56(m,1H),5.42-5.23(m,1H),5.23-5.01(m,2H),4.52-4.35(m,4H),4.04-3.93(m,2H),0.34(br s,6H).
19F NMR(376MHz,D
2O)-164.82,-200.86,-202.28.
31P NMR(162MHz,D
2O)δ96.94-91.91.
生物活性测试实验
实验例1：STING体外结合测试实验
荧光偏振测试法(fluorescence polarization assay,FP assay)被用于检测化合物对人STING蛋白的亲和力。反应体系中有一定量的荧光素标记的c-di-GMP和不同浓度的待测化合物，当加入重组人STING的C端蛋白，两种小分子与蛋白竞争性结合。结合态的荧光素标记的c-di-GMP在液相中转动较慢，此时检测到的荧光偏振程度也较高。荧光偏振程度与待测化合物浓度，亲和力呈反比关系。我们通过检测反应系中偏振光的大小，就可以精确地得知待测化合物对人STING的亲和力。
实验中用到的可溶性人STING蛋白序列是截取自人野生型内质网结合蛋白STING的C端部分，从140氨基酸至379氨基酸。人STING蛋白有多种序列差异的等位基因，不同等位基因对CDN亲和力不同(Yi,et.al.,“Single Nucleotide Polymorphisms of Human STING can affect innate immune response to cyclic dinucleotides"PLOS ONE.2013,8(10),e77846)。野生型STING序列(G230，R232，R293)约占了总体的57.9％。重组STING蛋白的N端是6His-SUMO序列，以利于蛋白正确折叠及纯化，经蛋白酶切除，C端STING用于FP测试。
FP测试使用384孔板，在每孔10μl反应体系中加有终浓度30nM的荧光素标记的c-di-GMP，10μM的人STING蛋白，和不同浓度的参照化合物或待测化合物。1000g离心1分钟，室温避光孵育30分钟，用Envision读板。
如上所述的STING体外结合测定实验结果如表1所示。
表1
| 化合物编号 | FP亲和力测试IC 50(μM) |
| 2',3'-cGAMP | 6.66 |
| 1A | 13.96 |
| 1B | 3.21 |
| 1C | 3.73 |
| 1D | 5.61 |
| 2A | 3.58 |
| 2B | 2.00 |
| 3A | 5.91 |
| 3B | 4.35 |
| 3C | 2.93 |
| 3D | 2.70 |
| 4A | 2.76 |
| 4B | 2.26 |
| 4C | 1.44 |
| 4D | 2.86 |
| 5A | 2.81 |
| 5B | 5.47 |
| 5C | 4.97 |
| 5D | 5.14 |
| 6A | 2.39 |
| 6B | 1.76 |
| 6C | 2.40 |
| 6D | 2.53 |
结论：在FP亲和力测试中，本发明化合物显示了高于内源性2'3'-cGAMP的对人野生型STING蛋白的亲和力。
实验例2：THP1-dual报告基因活性测试实验
测试所用THP1-Dual
TM细胞(InvivoGen目录代码：thpd-nfis)，是通过在人单核细胞系THP1中稳定整合两个诱导型报告基因构建。分泌型胚胎碱性磷酸酶(SEAP)报告基因的启动子序列组成包括一个IFN-β的基本启动子和上游的5个拷贝的NF-κB共表达转录应答元件(NF-κB consensus transcriptional response element)和3个拷贝的c-Rel结合位点。分泌性萤光素酶(Lucia)报告基因由5个干扰素刺激反应元件(interferon(IFN)-stimulated response elements)和一个ISG54的基本启动子驱动。从而使得同时研究STING的两个主要下游信号传导途径成为可能：通过检测SEAP活性研究NFκB途径：和通过评估Lucia荧光素酶的活性研究IRF途径。
用PB buffer(50mM HEPES,100mM KCl,3mM MgCl2,0.1mM DTT,85mM Sucrose,1mM ATP,0.1mM GTP,0.2％BSA)稀释好化合物。向96孔板中每孔添加20μL参照或待测化合物，随后添加180μL用PB buffer悬浮的THP1-Dual细胞(大约100,000个细胞/孔)。将平板在37℃，5％CO
2条件下孵育30分钟后，1000rpm离心10分钟，弃上清，用200μL/孔RPMI-1640洗两次，添加200μL每孔的RPMI-1640培养18小时。收集上清，根据制造商的说明使用QUANTI-Luc
TM定量IRF3途径的激活。
如上所述的THP1-dual体外结合测定结果如表2所示。
表2
| 化合物编号 | EC 50(μM) |
| 2',3'-cGAMP | 20.19 |
| ADU-S100 | 23.39 |
| 1A | 74.69 |
| 1B | 7.38 |
| 1C | 16.76 |
| 1D | 7.48 |
| 2A | 1.75 |
| 2B | 3.39 |
| 3B | 42.21 |
| 3C | 47.17 |
| 3D | 26.13 |
| 4A | 3.06 |
| 4B | 3.68 |
| 4C | 10.18 |
| 4D | 48.20 |
| 5A | 6.20 |
| 5B | 11.42 |
| 5C | 5.13 |
| 5D | 12.6 |
| 6A | 1.36 |
| 6B | 9.31 |
| 6C | 2.31 |
| 6D | 12.30 |
结论：在人单核细胞系THP-1中，本发明化合物具有很强的促β干扰素激活的能力。
实验例3：Raw-Dual报告基因活性测试实验
测试所用RAW-Dual
TM细胞(InvivoGen目录代码：rawd-ismip)，是通过在小鼠巨噬细胞系RAW264.7中稳定整合两个诱导型报告基因构建：通过检测SEAP活性研究NF-KB途径，和通过评估Lucia荧光素酶的活性研究IRF3途径。按每孔200μL将细胞(每孔50000细胞)悬液加入96孔板(康宁3599平底板)，37℃培养箱中培养18～24小时。第二天弃去培液，每孔加入200μL预先用培养基配置好的化合物溶液，室温孵育30分钟，吸去处理液，用无血清培养液洗两次，然后每孔加入200μL培养液，37℃培养箱中培养18～24小时。第三天每孔取20μL上清液，根据制造商的说明使用QUANTI-LucTM定量IRF3途径的激活。
如上所述的RAW细胞活性测定的结果如表3所示
表3
| 化合物编号 | Raw,EC50(μM) |
| ADU-S100 | 47.08 |
| 1B | 15.8 |
| 1C | 41.4 |
| 1D | 24.4 |
| 2A | 23.4 |
| 2B | 2.1 |
| 3C | 9.4 |
| 3D | 10.0 |
| 4B | 2.6 |
| 4C | 6.3 |
| 4D | 21.0 |
| 5B | 54.8 |
| 5C | 28.2 |
| 5D | 31.6 |
| 6B | 5.4 |
| 6C | 1.1 |
| 6D | 9.5 |
结论：测试中发现，在小鼠巨噬细胞系RAW报告基因测试实验中，本发明化合物具有很强的激活STING的能力。
实验例4：体内药效实验1
本实验通过4T1乳腺癌同系小鼠模型进行化合物药效评价。将1E5个4T1乳腺癌细胞(上海中科院细胞所)接种于6-8周龄的Balb/C小鼠(维通利华)皮下，当肿瘤体积达到100mm3后随机分组，每组8只。分组后的第1，第4，第8天依次进行瘤内给药。单次瘤内给药(IT)为分组后第1天。ADU-S100剂量组为100ug每只小鼠(单次)和30ug每只小鼠(三次)。化合物2B剂量组为30ug每只小鼠(单次),100ug每只小鼠(单次)和30ug每只小鼠(三次)。开始给药后进行肿瘤体积的测量，一周两次。肿瘤体积的计算公式为：V＝0.5a×b
2，a和b分别表示肿瘤的长径和短径。每个点为肿瘤体积的平均值和标准误(SEM)。组间差异采用two-way ANOVA进行统计学分析(第25天的统计学差异如图，Prism7,****p<0.0001)。给药组肿瘤体积显著小于对照组。化合物2B 100ug(单次)和30ug(三次)组肿瘤全部消失，肿瘤抑制效果优于同剂量的ADU-100。结果如图1所示。
实验例5：体内药效实验2
本实验通过CT-26结肠癌同系小鼠模型进行化合物药效评价。将3E5个CT-26结肠癌细胞(ATCC-CRL-2638)接种于6-8周龄的Balb/C小鼠(上海灵畅生物)皮下，当肿瘤体积达到100mm3后随机分组，每组8只。分组后的第1，第4，第8天依次进行瘤内给药，共三次。化合物2B每组的给药剂量分别为1ug每只小鼠,3ug每只小鼠,9ug每只小鼠,和18μg每只小鼠。化合物ADU-S100给药剂量为125μg每只 小鼠。开始给药后进行肿瘤体积的测量，一周三次。肿瘤体积的计算公式为：V＝0.5a×b
2，a和b分别表示肿瘤的长径和短径。每个点为肿瘤体积的平均值和标准误(SEM)。对照组与给药组的组间差异采用two-way ANOVA进行统计学分析(第11天的统计学差异如图，Prism7,****p<0.0001)。
与对照组相比，给药组小鼠肿瘤增长速度明显变慢。化合物2B对小鼠肿瘤生长的抑制作用呈现出剂量依赖性。化合物2B 18μg每只小鼠跟化合物ADU-S100 125μg每只小鼠产生的肿瘤抑制作用相当。结果如图2所示。
实验例6：体内药效实验3
本实验通过MC38结肠癌同系小鼠模型进行化合物药效评价。将3E5个MC38结肠癌细胞(南京科佰)接种于6-8周龄的C57BL/6小鼠(上海必凯)皮下，当肿瘤体积达到100mm3左右后随机分组，每组5-6只。分组后的第1，第4，第8天依次进行瘤内给药。单次瘤内给药为分组后第1天。ADU-S100剂量组为100ug每只小鼠(单次)。化合物2B剂量组为100ug每只小鼠(单次)，30ug每只小鼠(三次)和10ug每只小鼠(三次)。化合物6C剂量组为30ug每只小鼠(三次)。开始给药后进行肿瘤体积的测量，一周两次。肿瘤体积的计算公式为：V＝0.5a×b
2，a和b分别表示肿瘤的长径和短径。每个点为肿瘤体积的平均值和标准误(SEM)。对照组与给药组的组间差异采用two-way ANOVA进行统计学分析(第28天的统计学差异如图，Prism7,****p<0.0001)。
与对照组相比，给药组小鼠肿瘤生长明显受到抑制。化合物2B 30ug(三次)和10ug(三次)以及化合物6C 30ug(三次)组小鼠肿瘤全部消失。结果如图3所示。
Claims (19)
- 式(Ⅰ)所示化合物、其光学异构体及其药效上可接受的盐，
其中，R 1、R 1a分别独立地选自
T 1、T 2、T 3、T 4、T 5、T 6、T 7、T 8、T 9、T 10、T 11、T 12、T 13分别独立地选自-C(R)-和-N-；L 1、L 2分别独立地选自-O-、-N(R)-、-C(RR)-和-C(＝O)-；R分别独立地选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基任选被1、2或3个R’取代；
R’选自F、Cl、Br、I、OH、NH 2和CH 3；R 2、R 2a分别独立地选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2- 6炔基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基任选被1、2或3个R取代；R 3、R 3a分别独立地选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2- 6炔基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基任选被1、2或3个R取代；R 4、R 4a分别独立地选自H、卤素、OH、NH 2、CN、N 3、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基任选被1、2或3个R取代；R 5、R 5a分别独立地选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2- 6炔基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基任选被1、2或3个R取代；R 6、R 6a分别独立地选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2- 6炔基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基任选被1、2或3个R取代；R 7、R 7a分别独立地选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基任选被1、2或3个R取代；R 10、R 10a分别独立地选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基任选被1、2或3个R取代；或者，R 7和R 10连接在一起，形成一个C 3-6环烷基、C 3-6环稀基或C 3-6环炔基，所述C 3-6环烷基、C 3-6环稀基或C 3-6环炔基任选被1、2或3个R取代；R 7a和R 10a连接在一起，形成一个C 3-6环烷基、C 3-6环稀基或C 3-6环炔基，所述C 3-6环烷基、C 3-6环稀基或C 3-6环炔基任选被1、2或3个R取代；R 8选自BH 3 -和-S(R 9)；R 9选自H、CH 2OC(＝O)R 11、CH 2OC(＝O)OR 11、CH 2CH 2SC(＝O)R 11和CH 2CH 2SSCH 2R 11；R 11选自C 6-10芳基、5～10元杂芳基、C 1-6杂环烷基和C 1-20烷基，所述C 1-20烷基任选被1、2、3、4或5个C 6-10芳基、C 3-10环烷基、OH和F取代；或者，R 4与R 6或R 4a与R 6a连接在一起形成一个5～6元杂环烷基；X 1、X 1a分别独立地选自-NH-、-O-、-S-和-CH 2-；X 2、X 2a分别独立地选自-NH-、-O-、-S-和-CH 2-；X 3、X 3a分别独立地选自-O-和-S-；Y、Y a分别独立地选自-O-、-S-、-CH 2-和-C(＝CH 2)-；所述5～6元杂环烷基、5～10元杂芳基或C 1-6杂环烷基包含1、2或3个独立选自-O-、-NH-、-S-、-C(＝O)-、-C(＝O)O-、-S(＝O)-、-S(＝O) 2-和N的杂原子或杂原子团；且，当R 1或R 1a选自时，式(I)所示的化合物不选自
- 根据权利要求1所述化合物、其光学异构体及其药效上可接受的盐，其中，当R 8选自BH 3 -时，R 4和R 4a其中一个选自F、Cl和Br，另一个选自F、Cl、Br、OH、OCH 3或N 3。
- 根据权利要求1所述化合物、其光学异构体及其药效上可接受的盐，其中，R分别独立地选自H、卤素、 OH、NH 2、CN、C 1-3烷基、C 1-3烷氧基、C 1-3烷硫基和C 1-3烷氨基，其中C 1-3烷基、C 1-3烷氧基、C 1-3烷硫基和C 1-3烷氨基任选被1、2或3个R’取代。
- 根据权利要求3所述化合物、其光学异构体及其药效上可接受的盐，其中，R分别独立地选自H、F、Cl、Br、I、OH、NH 2、CN、Me、其中Me、
任选被1、2或3个R’取代。
- 根据权利要求4所述化合物、其光学异构体及其药效上可接受的盐，其中，R分别独立地选自H、F、Cl、Br、I、OH、NH 2、CN、Me、
- 根据权利要求1～5任意一项所述化合物、其光学异构体及其药效上可接受的盐，其中，R 1、R 1a分别独立地选自
- 根据权利要求6所述化合物、其光学异构体及其药效上可接受的盐，其中，R 1、R 1a分别独立地选自
- 根据权利要求1～5任意一项所述化合物、其光学异构体及其药效上可接受的盐，其中，R 2、R 2a、R 3、R 3a、R 5和R 5a、R 6和R 6a分别独立地选自H；任选地，R 6和R 6a分别独立地选自H和甲基。
- 根据权利要求1～5任意一项所述化合物、其光学异构体及其药效上可接受的盐，其中，R 4、R 4a分别独立地选自F、OH、NH 2、N 3和
- 根据权利要求1～5任意一项所述化合物、其光学异构体及其药效上可接受的盐，其中，R 7、R 7a分别独立地选自H和CH 3。
- 根据权利要求1～5任意一项所述化合物、其光学异构体及其药效上可接受的盐，其中，R 4与R 6连接在一起，结构单元选自
- 根据权利要求1～5任意一项所述化合物、其光学异构体及其药效上可接受的盐，其中，R 4a与R 6a连接在一起，结构单元选自
- 根据权利要求1所述化合物、其光学异构体及其药学上可接受的盐，其选自
其中，R 1、R 1a分别独立地选自
R 4、R 4a分别独立地选自H、卤素、OH、NH 2、CN、N 3、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基，其中C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基任选被1、2或3个R取代；R 6选自H、卤素、OH、NH 2、CN、C 1-6烷基、C 1-6烷氧基、C 1-6烷硫基、C 1-6烷氨基和C 2-6炔基，其中C 1- 6烷基、C 1-6烷氧基、C 1-6烷硫基和C 1-6烷氨基任选被1、2或3个R取代；或者，R 4与R 6连接在一起形成一个5～6元杂环烷基；R 8选自BH 3 -和-S(R 9)；R 9选自H、CH 2OC(＝O)R 11、CH 2OC(＝O)OR 11、CH 2CH 2SC(＝O)R 11和CH 2CH 2SSCH 2R 11；R 11选自C 6-10芳基、5～10元杂芳基、C 1-6杂环烷基和C 1- 20烷基，所述C 1- 20烷基任选被1、2、3、4或5个C 6-10芳基、C 3-10环烷基、OH和F取代；X 1、X 1a分别独立地选自-NH-、-O-、-S-和-CH 2-；X 2、X 2a分别独立地选自-NH-、-O-、-S-和-CH 2-；X 3、X 3a分别独立地选自-O-和-S-；Y、Y a分别独立地选自-O-、-S-、-CH 2-和-C(＝CH 2)-；当R 8选自BH 3 -时，R 4和R 4a其中一个选自F、Cl和Br，另一个选自F、Cl、Br、OH、OCH 3或N 3；当R 8选自-S(R 9)时，R 4和R 4a其中一个选自F、Cl和Br，另一个选自OH、OCH 3或N 3；或者，当R 8选自-S(R 9)，且R 4和R 4a其中一个选自F、Cl和Br，另一个不选自OH、OCH 3或N 3时，R 4和R 4a不选自且R 4和R 4a不同时选自
- 根据权利要求1～7、9或10任意一项所述化合物、其光学异构体及其药效上可接受的盐，其选自
其中，R 1、R 1a如权利要求1～7所定义；R 4a分别如权利要求1或9所定义；R 7、R 7a分别如权利要求1或10所定义；R 8、R 6a如权利要求1所定义。 - 根据权利要求14述化合物、其光学异构体及其药效上可接受的盐，其选自
- 根据权利要求1所述化合物、其光学异构体及其药学上可接受的盐，其选自
其中，R 1、R 1a分别独立地选自
X 1、X 1a分别独立地选自-NH-、-O-、-S-和-CH 2-；X 2、X 2a分别独立地选自-NH-、-O-、-S-和-CH 2-；X 3、X 3a分别独立地选自-O-和-S-；Y、Y a分别独立地选自-O-、-S-、-CH 2-和-C(＝CH 2)-。 - 根据权利要求16所述化合物、其光学异构体及其药学上可接受的盐，其选自
- 根据权利要求13～17任意一项所述化合物、其光学异构体及其药效上可接受的盐，其中，R 1、R 1a分别独立地选自
- 下式化合物、其光学异构体及其药效上可接受的盐，其选自
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| JP2021546037A JP7492523B2 (ja) | 2018-10-12 | 2019-10-12 | 環状ジヌクレオチド化合物及びその使用 |
| US17/284,154 US11401295B2 (en) | 2018-10-12 | 2019-10-12 | Cyclic dinucleotide compound and uses thereof |
| EP19871233.3A EP3868773A4 (en) | 2018-10-12 | 2019-10-12 | CYCLIC DINUCLEOTIDE COMPOUND AND ITS APPLICATIONS |
| CN201980062095.0A CN112867727B (zh) | 2018-10-12 | 2019-10-12 | 环二核苷酸类化合物及其应用 |
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| CN201811367721 | 2018-11-16 | ||
| CN201811367721.0 | 2018-11-16 | ||
| CN201910129734.2 | 2019-02-21 | ||
| CN201910129734 | 2019-02-21 | ||
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| EP (1) | EP3868773A4 (zh) |
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Cited By (9)
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| CN111909223A (zh) * | 2020-07-17 | 2020-11-10 | 清华大学 | 环二核苷酸共价修饰物及其制备方法和应用 |
| CN113045612A (zh) * | 2021-03-17 | 2021-06-29 | 常州市白云生物科技有限公司 | 3-O-苄基-4-C-羟甲基-1,2-O-异亚丙基-α-D-呋喃核糖的合成方法 |
| WO2021206158A1 (ja) | 2020-04-10 | 2021-10-14 | 小野薬品工業株式会社 | がん治療方法 |
| WO2022217022A1 (en) | 2021-04-10 | 2022-10-13 | Profoundbio Us Co. | Folr1 binding agents, conjugates thereof and methods of using the same |
| WO2022226317A1 (en) | 2021-04-23 | 2022-10-27 | Profoundbio Us Co. | Anti-cd70 antibodies, conjugates thereof and methods of using the same |
| WO2023280227A2 (en) | 2021-07-06 | 2023-01-12 | Profoundbio Us Co. | Linkers, drug linkers and conjugates thereof and methods of using the same |
| WO2025072406A1 (en) | 2023-09-26 | 2025-04-03 | Profoundbio Us Co. | Ptk7 binding agents, conjugates thereof and methods of using the same |
| WO2025149661A1 (en) | 2024-01-10 | 2025-07-17 | Genmab A/S | Slitrk6 binding agents, conjugates thereof and methods of using the same |
| WO2025181219A1 (en) | 2024-02-29 | 2025-09-04 | Genmab A/S | Egfr and c-met bispecific binding agents, conjugates thereof and methods of using the same |
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| KR102782200B1 (ko) * | 2017-08-30 | 2025-03-17 | 에이제이 사이언시즈 (이씽) 코., 엘티디 | 인터페론 유전자 조절인자의 자극제로서의 환식 다이-뉴클레오타이드 |
| CN116589510B (zh) * | 2023-04-20 | 2025-08-26 | 安徽大学 | 一种氟代苄基核糖中间体的制备方法与应用 |
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| WO2021206158A1 (ja) | 2020-04-10 | 2021-10-14 | 小野薬品工業株式会社 | がん治療方法 |
| CN111909223A (zh) * | 2020-07-17 | 2020-11-10 | 清华大学 | 环二核苷酸共价修饰物及其制备方法和应用 |
| CN113045612A (zh) * | 2021-03-17 | 2021-06-29 | 常州市白云生物科技有限公司 | 3-O-苄基-4-C-羟甲基-1,2-O-异亚丙基-α-D-呋喃核糖的合成方法 |
| WO2022217022A1 (en) | 2021-04-10 | 2022-10-13 | Profoundbio Us Co. | Folr1 binding agents, conjugates thereof and methods of using the same |
| WO2022226317A1 (en) | 2021-04-23 | 2022-10-27 | Profoundbio Us Co. | Anti-cd70 antibodies, conjugates thereof and methods of using the same |
| WO2023280227A2 (en) | 2021-07-06 | 2023-01-12 | Profoundbio Us Co. | Linkers, drug linkers and conjugates thereof and methods of using the same |
| WO2025072406A1 (en) | 2023-09-26 | 2025-04-03 | Profoundbio Us Co. | Ptk7 binding agents, conjugates thereof and methods of using the same |
| WO2025149661A1 (en) | 2024-01-10 | 2025-07-17 | Genmab A/S | Slitrk6 binding agents, conjugates thereof and methods of using the same |
| WO2025181219A1 (en) | 2024-02-29 | 2025-09-04 | Genmab A/S | Egfr and c-met bispecific binding agents, conjugates thereof and methods of using the same |
Also Published As
| Publication number | Publication date |
|---|---|
| US20210340169A1 (en) | 2021-11-04 |
| CN112867727A (zh) | 2021-05-28 |
| JP7492523B2 (ja) | 2024-05-29 |
| US11401295B2 (en) | 2022-08-02 |
| EP3868773A4 (en) | 2022-07-13 |
| EP3868773A1 (en) | 2021-08-25 |
| CN112867727B (zh) | 2024-05-17 |
| JP2022508786A (ja) | 2022-01-19 |
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