WO2020043154A1 - 作为免疫调节剂的联苯化合物及其用途 - Google Patents
作为免疫调节剂的联苯化合物及其用途 Download PDFInfo
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- WO2020043154A1 WO2020043154A1 PCT/CN2019/103225 CN2019103225W WO2020043154A1 WO 2020043154 A1 WO2020043154 A1 WO 2020043154A1 CN 2019103225 W CN2019103225 W CN 2019103225W WO 2020043154 A1 WO2020043154 A1 WO 2020043154A1
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- 0 Cc1cccc(-c2cccc(COc3ccc(C*(*)*)c(N)c3)c2NC)c1* Chemical compound Cc1cccc(-c2cccc(COc3ccc(C*(*)*)c(N)c3)c2NC)c1* 0.000 description 2
- JSSOHCLMJVFQKE-QPJJXVBHSA-N CC1OC(C)CN(CC/C=C/c2c(C)c(Br)ccc2)C1 Chemical compound CC1OC(C)CN(CC/C=C/c2c(C)c(Br)ccc2)C1 JSSOHCLMJVFQKE-QPJJXVBHSA-N 0.000 description 1
- NWLOPXPIJPLDHB-UNQAUBFISA-N C[C@H]([C@@H](C(O)=O)NCc(c(C)c1)cc(Cl)c1OCc1c(C)c(-c2cccc(/C=C/CCN(CC3)CC3(F)F)c2C)ccc1)O Chemical compound C[C@H]([C@@H](C(O)=O)NCc(c(C)c1)cc(Cl)c1OCc1c(C)c(-c2cccc(/C=C/CCN(CC3)CC3(F)F)c2C)ccc1)O NWLOPXPIJPLDHB-UNQAUBFISA-N 0.000 description 1
- YICFABVCTMYCBO-VZUCSPMQSA-N Cc(c(C=O)c1)cc(OCc2cccc(-c3cccc(/C=C/CCN4CCCCC4)c3C)c2C)c1Cl Chemical compound Cc(c(C=O)c1)cc(OCc2cccc(-c3cccc(/C=C/CCN4CCCCC4)c3C)c2C)c1Cl YICFABVCTMYCBO-VZUCSPMQSA-N 0.000 description 1
- ZJXXWSUWHLABKL-WEVVVXLNSA-N Cc(c(CNCC(OC)=O)c1)cc(OCc2c(C)c(-c3cccc(/C=C/CCN(CC4)CC4(F)F)c3C)ccc2)c1Cl Chemical compound Cc(c(CNCC(OC)=O)c1)cc(OCc2c(C)c(-c3cccc(/C=C/CCN(CC4)CC4(F)F)c3C)ccc2)c1Cl ZJXXWSUWHLABKL-WEVVVXLNSA-N 0.000 description 1
- NBQLIOYOFMKKDQ-OYLQIYHASA-N Cc1c(/C=C/CCN(CCC2)C[C@@H]2O)cccc1Br Chemical compound Cc1c(/C=C/CCN(CCC2)C[C@@H]2O)cccc1Br NBQLIOYOFMKKDQ-OYLQIYHASA-N 0.000 description 1
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- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
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- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/4025—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil not condensed and containing further heterocyclic rings, e.g. cromakalim
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- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
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- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
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- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
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- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
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- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
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- A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
- A61K31/541—Non-condensed thiazines containing further heterocyclic rings
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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- C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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- C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
- C07D295/08—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms
- C07D295/096—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms separated by carbocyclic rings or by carbon chains interrupted by carbocyclic rings
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- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
Definitions
- the present invention relates to a biphenyl compound capable of inhibiting PD-1 / PD-L1 protein / protein interaction, a preparation method thereof, and use thereof in preparing a medicament for treating or preventing a disease related to the signal pathway.
- immune checkpoint inhibitors represented by PD-1 / PD-L1 antibodies have made significant clinical research progress in the field of Cancer immunotherapy. Unlike traditional chemotherapy based on cytotoxicity, tumor immunotherapy stimulates the body's immune system and enhances its own anti-tumor immunity, thereby achieving the purpose of persistently suppressing or killing tumor cells, which provides a new weapon for fighting cancer (Sharma P., Allison JP, 2015, Science, 348: 56-61).
- PD-1 Programmed death receptor 1
- CD28 CD134 (OX40), Glucocorticoid-induced TNFR-related protein (GITR), CD137, CD27, HVEM and other activating receptors that transmit activation signals, PD-1 and ligands
- ITIM immunoreceptor tyrosine-based inhibitory motif
- the immunoreceptor tyrosine-based inhibitory motif (ITIM) of the intracellular segment is activated to activate protein phosphatase (SH-2domain containing protein tyrosine phosphatase 1/2, SHP-1 / 2) Immune-suppressive signals are transmitted to the cell, thereby blocking the continued activation of T cells (Cheng X., Veverka V., Radhakrishnan A., et al. 2013, J. Biol. Chem., 288: 11771-11785 ).
- PD-1 ligands include programmed death ligand 1 (PD-L1) and programmed death ligand 2 (PD-L2), of which PD-L1 can be Persistently expressed in a variety of tumor cells, T cells, APC and a variety of non-hematopoietic cells; PD-L2 is restrictedly expressed in dendritic cells and macrophages. Under normal physiological conditions, the interaction of PD-1 and PD-L2 can inhibit T cell activation, which is essential for maintaining immune tolerance in the normal body; on the other hand, it can prevent tissue damage caused by excessive activation of the immune response.
- PD-L1 programmed death ligand 1
- PD-L2 programmed death ligand 2
- T cell activation which is essential for maintaining immune tolerance in the normal body; on the other hand, it can prevent tissue damage caused by excessive activation of the immune response.
- small-molecule compounds Compared with biological macromolecules, small-molecule compounds have many advantages such as oral administration, convenient administration, and significantly lower prices. Therefore, finding small-molecule PD-1 / PD-L1 inhibitors has great theoretical significance and application value.
- the compound of formula I according to the present invention can selectively inhibit PD-1 and / or PD-L1, and can be used to treat and / or prevent diseases mediated by the PD-1 / PD-L1 signaling pathway.
- diseases include, but are not limited to, cancer, autoimmune diseases, infectious diseases, and sepsis.
- the present invention provides a biphenyl compound represented by Formula I having PD-1 and / or PD-L1 inhibitory activity.
- the compounds of formula I described herein include their pharmaceutically acceptable salts.
- the invention provides a method for preparing a compound of formula I according to the invention.
- the invention also provides the use of a compound of formula I of the invention in the manufacture of a medicament for use as a PD-1 and / or PD-L1 inhibitor.
- the present invention also provides the use of a compound of formula I of the present invention in the preparation of a medicament for treating or preventing cancer, autoimmune disease, infectious disease and other diseases.
- the present invention also provides a pharmaceutical composition
- a pharmaceutical composition comprising a compound of formula I (including a pharmaceutically acceptable salt thereof) according to the present invention and an optional pharmaceutically acceptable excipient or carrier.
- R 1 is selected from hydrogen, C 1 -C 3 alkyl, CN, halogen, C 1 -C 3 haloalkyl;
- R 2 is selected from hydrogen, C 1 -C 3 alkyl, CN, halogen, C 1 -C 3 alkoxy;
- L is optionally substituted C 3 -C 6 alkenyl group, a C 3 -C 6 alkenyl group optionally substituted with a CH 2 -C (O) -, - S (O) - or -S (O ) 2 -replaced by;
- B is hydrogen, or NR 6 R 7 , or an optionally substituted 4- to 7-membered saturated or partially unsaturated heterocyclic ring having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
- R 6 and R 7 are independently hydrogen, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, hydroxy C 1 -C 3 alkyl, or carboxy C 1 -C 3 alkyl;
- R 8 is selected from hydrogen, C 3 -C 6 alkenyl, halogenated C 1 -C 4 alkyl, hydroxy C 1 -C 4 alkyl,-(CH 2 ) n X and-(CH 2 ) n Ar;
- n 1, 2, 3, or 4;
- X is selected from hydrogen, -CH 3 , -CF 3 , C 1 -C 4 alkoxy, -N (CH 3 ) 2 , C 3 -C 6 cycloalkyl optionally substituted with one or two halogens,- CN, -CO 2 Rg, -C (O) NH 2 , -C (O) N (CH 3 ) 2 , Morpholine, tetrahydropyranyl, pyrrolidinyl optionally substituted with hydroxy, and piperidinyl optionally substituted with one or two groups independently selected from: C 1 -C 4 alkyl, carboxyl, hydroxy And C 1 -C 4 alkoxycarbonyl;
- Rg is selected from hydrogen and C 1 -C 4 alkyl
- Ar is selected from benzodioxanyl, indazolyl, isoquinolinyl, isoxazolyl, naphthyl, oxadiazolyl, phenyl, pyridyl, pyrimidinyl, and quinolinyl; wherein each ring is optionally Substituted by 1, 2, 3 or 4 substituents independently selected from the group consisting of: C 1 -C 4 alkoxy, C 1 -C 4 alkoxycarbonyl, C 1 -C 4 alkoxycarbonylamino, C 1 -C 4 alkyl, C 1 -C 4 alkylcarbonyl, C 1 -C 4 alkylsulfonyl, formamido, formamido C 1 -C 4 alkyl,-(CH 2 ) q CO 2- ( C 1 -C 4 alkyl),-(CH 2 ) q OH, carboxyl, cyano, formyl, halogen, halogenated C 1
- t 0, 1, 2, 3, or 4;
- z is 1, 2 or 3;
- Each R z is independently selected from C 1 -C 4 alkoxy, C 1 -C 4 alkoxycarbonyl, C 1 -C 4 alkoxycarbonyl C 1 -C 4 alkyl, C 1 -C 4 alkyl, C 1 -C 4 alkylamido, C 1 -C 4 alkylamino, C 1 -C 4 alkylcarbonyl, amido, carboxyl, carboxyl C 1 -C 4 alkyl, di (C 1 -C 4 alkyl Alkyl) amido, di (C 1 -C 4 alkyl) amino, halogen, halogenated C 1 -C 4 alkoxy, halogenated C 1 -C 4 alkyl, hydroxyl, hydroxyl C 1 -C 4 alkyl , Morpholinyl, -NR c R d , (NR c R d ) C 1 -C 4 alkyl, -NR e R f , (NR e R
- R c and Rd are independently selected from hydrogen, C 2 -C 4 alkenylcarbonyl, C 1 -C 4 alkoxycarbonyl, C 1 -C 6 alkyl, C 1 -C 4 alkylcarbonyl, amido C 1 -C 4 alkyl, amino C 1 -C 4 alkyl, aryl C 1 -C 4 alkyl, C 3 -C 10 cycloalkyl, (C 3 -C 10 cycloalkyl) C 1 -C 4 alkyl Alkyl, halo C 1 -C 4 alkylcarbonyl, heterocyclyl C 1 -C 4 alkyl, heterocyclyl C 1 -C 4 alkylcarbonyl, hydroxyl C 1 -C 6 alkyl, and hydroxyl C 1 -C 4 -alkylcarbonyl, wherein the amido C 1 -C 4 alkyl, the amino C 1 -C 4 alkyl, the aryl C 1 -C 4
- R e and R f together with the atom to which they are attached form a group selected from morpholine and Ring
- R 3 is independently selected from C 2 -C 4 alkenyl, C 1 -C 4 alkoxy, C 1 -C 4 alkyl, cyano, halogen, and halogenated C 1 -C 4 alkyl;
- n 0, 1 or 2;
- R 4 is selected from hydrogen, C 1 -C 4 alkyl, benzyl, (C 3 -C 6 cycloalkyl) C 1 -C 3 alkyl, halo C 1 -C 4 alkyl, optionally via a second Hydroxy-substituted hydroxy C1-C6 alkyl and optionally cyano-substituted pyridyl (C 1 -C 3 alkyl); and
- R 5 is selected from hydrogen, C 1 -C 4 alkyl,-(CH 2 ) n N (CH 3 ) 2 , carboxy C 2 -C 6 alkenyl, carboxy C 1 -C 6 alkyl, and hydroxyl C 1 -C 6 alkyl, wherein the carboxy C 1 -C 6 alkyl and the alkyl portion of the hydroxy C 1 -C 6 alkyl are optionally substituted with a hydroxy or phenyl group, wherein the phenyl group is further optionally substituted with a hydroxy group; or R 5 is selected from the following groups:
- R w is -CONH 2 ;
- R 10 is selected from hydrogen, benzyl and methyl
- Each R 10 ′ is independently selected from hydrogen and C 1 -C 3 alkyl
- R 11 is selected from hydrogen, C 1 -C 3 alkyl and benzyl
- R 12 is selected from C 2 -C 4 alkenyl and C 1 -C 4 alkyl
- R 50 is selected from hydrogen, C 1 -C 6 alkyl and C 1 -C 6 alkoxycarbonyl;
- R 4 and R 5 together with the nitrogen atom to which they are attached form a ring selected from:
- s 0, 1, or 2;
- z is 1, 2 or 3;
- p 0, 1 or 2;
- R 13 and R 13 ′ are independently selected from hydrogen, carboxyl, hydroxy C 1 -C 4 alkyl, oxo and —C (O) NHSO 2 R 16 ;
- R 14 is independently selected from hydrogen, hydroxy C 1 -C 4 alkyl, oxo and carboxyl;
- Each R 15 is independently selected from C 1 -C 4 alkoxycarbonyl, C 1 -C 6 alkyl, carboxyl, halogen, hydroxyl, hydroxyl C 1 -C 4 alkyl, -NR c ' R d', and phenyloxy Carbonyl, wherein the phenyl is optionally substituted by nitro, wherein R c ′ and R d ′ are independently selected from hydrogen, C 1 -C 4 alkoxycarbonyl, and C 1 -C 4 alkylcarbonyl; and
- R 16 is selected from trifluoromethyl, cyclopropyl, C 1 -C 4 alkyl, dimethylamino, and methyl-substituted imidazolyl.
- R 1 and R 2 are independently selected from hydrogen and C 1 -C 3 alkyl
- n 1, 2, 3, or 4;
- Ar is selected from benzodioxanyl, indazolyl, isoquinolinyl, isoxazolyl, naphthyl, oxadiazolyl, phenyl, pyridyl, pyrimidinyl, and quinolinyl; wherein each ring is optionally Substituted by 1, 2, 3 or 4 substituents independently selected from the group consisting of: C 1 -C 4 alkoxy, C 1 -C 4 alkoxycarbonyl, C 1- C 4 alkoxycarbonylamino, C 1 -C 4 alkyl, C 1 -C 4 alkylcarbonyl, C 1 -C 4 alkylsulfonyl, amido, amido C 1 -C 4 alkyl,-(CH 2 ) q CO 2 C 1 -C 4- alkyl,-(CH 2 ) q OH, carboxyl, cyano, formyl, halogen, halogenated C 1 -C 4 alky
- R 3 is independently selected from C 1 -C 4 alkoxy and halogen.
- R 1 and R 2 are independently selected from methyl and halogen
- Ar is pyridyl optionally substituted with one or two groups independently selected from C 1 -C 4 alkyl, C 1 -C 4 alkylsulfonyl, formylamino, cyano, and halogen;
- R 3 is halogen
- halogen is fluorine, chlorine, bromine or iodine.
- alkyl in the present invention refers to having 1 to about 20 carbon atoms, usually 1 to 12 carbon atoms, preferably 1 to 8, 1 to 6, even more preferably 1 to 4 carbons.
- straight-chain alkyl groups include those having 1 to 8 carbon atoms, such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl; branched alkyl groups Examples include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, isohexyl, and 2,2-dimethylpropyl; examples of cycloalkyl include but It is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
- haloalkyl includes monohaloalkyl and polyhaloalkyl (wherein all halogen atoms may be the same or different).
- a partially halogenated alkyl is a "haloalkyl" within the meaning herein.
- Examples of the halogenated alkyl group include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3-difluoropropyl, and the like.
- alkoxy group in the present invention refers to a group formed by connecting the above-mentioned alkyl group with an oxygen atom, wherein the oxygen atom has a free bonding ability, such as a methoxy group, an ethoxy group, a propoxy group, and a butyl group.
- pharmaceutically acceptable salt refers to pharmaceutically acceptable acid and base addition salts and solvates.
- substituted means that an organic group (containing one or more bonds bonded to a hydrogen atom) as defined herein is substituted with one or more bonds bonded to a non-hydrogen atom or a group of atoms,
- the non-hydrogen atom or atomic group is a substituent.
- the substituents of the compounds of the present invention include, for example, halogen, alkyl (preferably C 1-8 alkyl, C 1-6 alkyl or C 1-4 alkyl), alkoxy (preferably C 1-8 alkoxy, C 1-6 alkoxy or C 1-4 alkoxy), haloalkyl (preferably C 1-8 haloalkyl, C 1-6 haloalkyl or C 1-4 haloalkyl), haloalkoxy (preferably C 1- 8 haloalkoxy, C 1-6 haloalkoxy or C 1-4 haloalkoxy), hydroxy, hydroxyalkyl (preferably hydroxy C 1-8 alkyl, hydroxy C 1-6 alkyl, or hydroxy C 1-4 alkyl).
- alkyl preferably C 1-8 alkyl, C 1-6 alkyl or C 1-4 alkyl
- alkoxy preferably C 1-8 alkoxy, C 1-6 alkoxy or C 1-4 alkoxy
- haloalkyl preferably C 1-8
- the invention provides a method for preparing a compound of formula I of the invention:
- a compound of formula I is formed by reductive amination under the action of an organic solvent and a reducing agent, wherein R 1 , R 2 , R 3 , R 4 , R 5 , L, B, Z and m are as previously defined.
- the compound of formula F may be an amino acid or an amino alcohol or an ester of an amino acid.
- the compound of formula A and formula F are reductively aminated to obtain the ester of compound of formula I, and the compound of formula I is obtained by hydrolysis.
- the reducing agent used in the above-mentioned reductive amination reaction may be sodium borohydride (NaBH 4 ), sodium cyanoborohydride (NaBH 3 CN), sodium acetate borohydride (NaBH 3 OAc), etc .
- the acid catalyst used may be acetic acid, hydrochloric acid, Trifluoroacetic acid
- the basic catalyst used can be TEA, DIPEA, etc .
- the desiccant used can be anhydrous magnesium sulfate (MgSO 4 ), anhydrous sodium sulfate (Na 2 SO 4 ), and molecular sieve; etc .
- the solvent used can be CH 3 OH, CH 2 Cl 2 and 1,2-dichloroethane.
- the invention also provides the use of the compound of formula A as an intermediate for preparing the compound of formula I according to the invention.
- Scheme 1 includes the following steps (a)-(c):
- the above-described nucleophilic substitution reaction (a) is NaOH, Na2CO 3, K 2 CO 3 as base to acid and the like, the reaction temperature is 25 ⁇ 140 °C, the reaction time is 2 to 72 hours.
- the solvents used in the reaction are commonly used solvents, such as water, methanol, ethanol, acetonitrile, benzene, xylene, acetone, N, N'-dimethylformamide, DMSO, and the like.
- the Suzuki cross-coupling reaction (b) uses palladium as a catalyst, such as Pd (PPh 3 ) 4 , Pd (dffp) Cl 2 , Pd (OAc) 2 , Pd (dba) 3 / PCy 3 , PdCl 2 , Pd (PPh 3 ) 4 Cl 2 and the like.
- a catalyst such as Pd (PPh 3 ) 4 , Pd (dffp) Cl 2 , Pd (OAc) 2 , Pd (dba) 3 / PCy 3 , PdCl 2 , Pd (PPh 3 ) 4 Cl 2 and the like.
- Ni as catalyst, such as NiCl 2 (dffp), NiCl 2 (dffp) / Zn, NiCl 2 (dffp) / BuLi, NiCl 2 (PPh 3 ) 2 / PPh 3 , NiCl 2 (NEt 3 ) 2 , NiCl 2 (NEt 3 ) 2 , NiCl (bipy), Ni (TPPS) 3 , Ni (COD) 2 , NiCl 2 (PPh 3 ) 2 / n-BuLi, Ni ⁇ P (OMe) 3 ⁇ 2 Cl 2 , NiCl 2 ( PCy 3 ) 2 and so on.
- the reaction is preferably performed in the presence of a base, such as KOAc, K 3 PO 4 , K 2 CO 3 , NaOH, Ba (OH) 2 , Na 2 CO 3 , CsF, or NaHCO 3 and the like.
- a base such as KOAc, K 3 PO 4 , K 2 CO 3 , NaOH, Ba (OH) 2 , Na 2 CO 3 , CsF, or NaHCO 3 and the like.
- the reaction temperature is 25 to 140 ° C, and the reaction time is 4 to 72 hours.
- the solvents used in the reaction are commonly used solvents, such as ethanol, THF, isopropanol, DMSO, DMF, dioxane, toluene, water and DME.
- the hydroxyl group is converted into a good leaving group, such as -OMs, -OTs or bromine, and triethylamine and DIPEA organic bases are used as deacidifying agents.
- the reaction temperature is 0 to 90 ° C.
- the time is 2 to 72 hours.
- the solvents used in the reaction are commonly used solvents, such as water, methanol, ethanol, isopropanol, tetrahydrofuran, dichloromethane, acetone, etc .; in the second step of the O-hydrocarbonation reaction of phenol, bases such as NaOH, K 2 CO 3 , KOH are used to remove the acid Agent, the reaction temperature is 0 to 90 ° C, and the reaction time is 2 to 72 hours.
- the solvents used in the reaction are commonly used solvents, such as water, methanol, ethanol, isopropanol, acetone, DMF, DMSO, benzene or xylene;
- Option two includes the following steps (d)-(e):
- the Mitsunobu reaction (d) can use nBu 3 P, PPh 3 , DIAD or DEAD as the activating reagent, the reaction temperature is -78 ° C to room temperature, and the reaction time is 2 to 72 hours.
- the solvent used in the reaction may be a common low-polarity solvent, such as THF, diethyl ether, dichloromethane, toluene, ethyl acetate, acetonitrile, DMF, and the like.
- the Suzuki cross-coupling reaction (e) can use palladium as a catalyst, such as Pd (PPh 3 ) 4 , Pd (dffp) Cl 2 , Pd (OAc) 2 , Pd (dba) 3 / PCy 3 , PdCl 2 , Pd ( PPh 3 ) 4 Cl 2 etc.
- a catalyst such as Pd (PPh 3 ) 4 , Pd (dffp) Cl 2 , Pd (OAc) 2 , Pd (dba) 3 / PCy 3 , PdCl 2 , Pd ( PPh 3 ) 4 Cl 2 etc.
- Ni as catalyst such as NiCl 2 (dffp), NiCl 2 (dffp) / Zn, NiCl 2 (dffp) / BuLi, NiCl 2 (PPh 3 ) 2 / PPh 3 , NiCl 2 (NEt 3 ) 2 , NiCl 2 (NEt 3 ) 2 , NiCl (bipy), Ni (TPPS) 3 , Ni (COD) 2 , NiCl 2 (PPh 3 ) 2 / n-BuLi, Ni (P (OMe) 3 ⁇ 2 Cl 2 , NiCl 2 (PCy 3 ) 2 and the like.
- the reaction can be performed in the presence of a base such as KOAc, K 3 PO 4 , K 2 CO 3 , NaOH, Ba (OH) 2 , Na 2 CO 3 , CsF, or NaHCO 3 and the like, and the reaction temperature is 25 to 140 ° C.
- the reaction time is 4 to 72 hours.
- the solvent used in the reaction may be a common solvent, such as ethanol, THF, isopropanol, DMSO, DMF, dioxane, toluene, water, DME, and the like.
- the compound of formula I can be purified by common separation methods, such as extraction, recrystallization, column chromatography and the like.
- reaction was stirred at room temperature for 16h.
- a 1N aqueous hydrochloric acid solution was slowly added to the reaction solution, the pH of the reaction solution was adjusted to about 5, and then diluted with dichloromethane (40 mL).
- the mixture was washed once with water (40 mL), and the organic phase was dried over anhydrous Na 2 SO 4 and then filtered.
- reaction solution was concentrated to obtain a crude product, which was purified by silica gel column chromatography (EA) to obtain an off-white solid (E) -5-chloro-4-((3 '-(4- (piperidin-1-yl) butan-1- En-1-yl) -2,2'-dimethyl- [1,1'-diphenyl] -3-yl) methoxy) -2-methylbenzaldehyde (262 mg, 0.523 mmol, 96% ).
- (E) -1- (4- (3-bromo-2-methylphenyl) but-3-en-1-yl) thiomorpholine-1,1-dioxide (815.0mg, 2.283 mmol, 69%) is composed of (E) -1-bromo-3- (4-bromobut-1-en-1-yl) -2-methylbenzene (1.0 g, 3.311 mmol) and thiomorpholine -1,1-dioxide (894.0 mg, 6.622 mmol) was prepared by a similar procedure as in Example 6.
- Methanesulfonyl chloride (8.07 g, 70.8 mmol) was added to a dichloromethane solution of 5- (hydroxymethyl) nicotinonitrile (7.16 g, 53 mmol) and triethylamine (10.7 g, 106 mmol) at 0 ° C.
- the mixed solution was stirred at room temperature for 4 hours, and then 300 mL of a saturated aqueous ammonium chloride solution was added.
- the solution was washed with dichloromethane (200mL) and extracted three times, the organic phases were combined, dried over anhydrous Na 2 SO 4 and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was used in the next step without purification.
- Tetrahydrofuran was removed, diluted with water (5mL), the pH was adjusted to weakly acidic, a white solid was precipitated, filtered and dried to obtain (E) -1- (5-chloro-4-((2,2'-dimethyl-3 '-(4-morpholinbut-1-en-1-yl)-[1,1'-di Phenyl] -3-yl) methoxy) -2-methylbenzyl) 3-hydroxypiperidine-2-carboxylic acid (35 mg, yield 22.2%).
- the detection method is used for evaluating the in vitro activity of the compound of the present invention, and includes an in vitro evaluation method of protein level binding inhibitory activity and a cell-level biological function activity evaluation method.
- the purpose of this test is to comprehensively evaluate the characteristics of PD-1 / PD-L1 binding inhibitory activity of different compounds and the biological activity of cell models, including the growth activity of specific model cells and biological activity of primary T cells.
- Example A In vitro PD-1 / PD-L1 binding inhibition screening method
- HTRF Homogeneous time-resolved fluorescence
- Human recombinant PD-1 protein with His tag (Cat #: 80448-R08H-100) and human recombinant PD-L1-Fc fusion protein (Cat #: 90251-C02H-100) were purchased from Sino Biological Inc. ) Company, anti-hFc-Eu 3+ and anti-His-XL665 antibodies were purchased from Cisbio, and other related reagents such as dilution buffer and detection buffer were purchased from Cisbio.
- the fluorescence detection instrument Tecan (Spark 10M) was purchased from Tecan, Switzerland.
- the experimental procedure was performed according to the procedure required for the instruction manual of the detection reagent (Invitrogen). The process is as follows:
- a control group is also provided for the detection reaction, including a 0-inhibition positive control without adding a test compound and a negative control without adding PD-1 protein. All tests were performed using duplicate wells.
- the fluorescence signal Tecan (Spark 10M) was used to detect the fluorescence signal of each well.
- the excitation wavelength was 320 nm and the detected emission wavelengths were 620 nm and 665 nm.
- inhibition rate (%) [1-(ratio of fluorescence signal of detection well-negative control) / (ratio of fluorescence signal of inhibition positive control-negative control) ⁇ 100%. After different concentrations of test compound were calculated binding inhibition rate, 50% inhibitory concentration is further calculated (IC 50). The data are shown in Table 2 below.
- Table 2 Representative compounds of the invention inhibits PD-1 PD-L1 binding IC / 50 data
- Compound IC 50 (nM) Compound IC 50 (nM) Compound IC 50 (nM) Compound IC 50 (nM) 5 120 10 14 13 29 16 15 19 17 twenty two 20 twenty three 4 25 36 27 2 29 3 30 56 33 27 36 12 41 11 47 38
- the compounds of the present invention have good in vitro PD-1 / PD-L1 binding inhibitory activity.
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Abstract
公开了涉及式I所示的联苯类化合物、其制备方法及其制药用途。式I中各基团的定义如说明书所述。所述化合物能够阻断PD-1/PD-L1信号通路之间的相互作用，可用于治疗或预防与该信号通路相关的疾病，例如癌症、自身免疫性疾病或慢性感染性疾病。 (I)
Description
交叉引用
本申请要求于2018年8月31日提交到中国专利局的发明名称为“作为免疫调节剂的联苯化合物及其用途”的中国专利申请201811013956.X的优先权，其内容通过引用以整体并入本文。
本发明涉及能够抑制PD-1/PD-L1蛋白/蛋白相互作用的联苯类化合物、其制备方法及其在在制备用于治疗或预防与该信号通路相关的疾病的药物中的用途。
近年来，以PD-1/PD-L1抗体为代表的免疫检查点抑制剂在肿瘤免疫治疗(Cancer immunotherapy)领域取得了重大的临床研究进展。与传统基于细胞毒作用的化疗不同，肿瘤免疫治疗通过激发人体的免疫系统，增强自身的抗肿瘤免疫力，从而达到持久抑制或杀死肿瘤细胞的目的，这为抗击癌症提供了新型武器(Sharma P.,Allison J.P.,2015,Science,348:56-61)。
程序性死亡受体1(Programmed death receptor 1，PD-1)主要表达于激活的T细胞、B细胞及骨髓细胞中，属于CD28家族。与CD28、CD134(OX40)、糖皮质激素诱导的TNFR相关蛋白(Glucocorticoid-induced TNFR-related protein,GITR)、CD137、CD27、HVEM等传递激活信号的活化性受体不同，PD-1与配体结合后通过活化胞内段的免疫受体酪氨酸抑制性基序(Immunoreceptor tyrosine-based inhibitory motif,ITIM)后激活蛋白磷酸水解酶(SH-2domain containing protein tyrosine phosphatase 1/2,SHP-1/2)等向胞内传递免疫抑制性信号，从而阻断T细胞的持续活化(Cheng X.,Veverka V.,Radhakrishnan A.,et al.2013,J.Biol.Chem.,288:11771-11785)。
PD-1的配体包括程序性死亡配体1(Programmed death protein ligand 1,PD-L1)和程序性死亡配体2(Programmed death protein ligand 2,PD-L2)两种，其中PD-L1可持续性表达于多种肿瘤细胞、T细胞、APC及多种非造血细胞；PD-L2只限制性地表达于树突状细胞和巨噬细胞中。在正常生理条件下，PD-1与PD-L2的相互作用会抑制T细胞激活，这对于维持正常机体的免疫耐受至关重要；另一方面可避免免疫反应过度激活导致组织损伤。然而，在肿瘤细胞中和病毒感染时，PD-L1的表达上调，同时T细胞上的PD-1被诱导性高表达， 导致PD-1信号通路持续激活而抑制T细胞增殖，造成肿瘤细胞和病原体的免疫逃逸(Fuller M.J.,Callendret B.,Zhu B.,et al.2013,Proc.Natl.Acad.Sci.USA.,110:15001-15006；Dolan D.E.,Gupta S.,2014,CancerControl,21:231-237；Chen L.,Han X.,2015,J.Clin.Invest.,125:3384-3391；Postow M.A.,Callahan M.K.,Wolchok J.D.,2015,J.Clin.Oncol.,33:1974-1982)。因此，需要通过激活共刺激信号(踩“油门”)并抑制共抑制信号(松“刹车”)而重新激活T细胞攻击肿瘤细胞，进而实现肿瘤免疫治疗。临床研究表明，免疫检查点阻断是T细胞激活的关键策略之一，而几个抗体药物的成功上市和巨大的市场潜力无疑使免疫检查点的研究成为了各大制药公司的聚焦点(Pardoll D.M.,2012,Nat.Rev.Cancer.,12:252-264)。
目前，针对阻断这一信号通路已有5个抗体药物获批上市，分别是MSD的Pembrolizumab(PD-1，2014)，BMS的Nivolumab(PD-1，2014)，罗氏的Atezolizumab(PD-L1，2016)，新基的Durvalumab(PD-L1，2017)和辉瑞的Avelumab(PD-L1，2017)。多项临床研究证实这些抗体药物在多种肿瘤中有效，如黑色素瘤、非小细胞肺癌、肾细胞癌、尿路上皮癌、宫颈癌、胃癌、头颈癌、及霍奇金淋巴瘤等。
与基于大分子的抗体的PD-1/PD-L1抑制剂相比，小分子的PD-1/PD-L1抑制剂目前还未得到人们的充分重视，还处于前期研发阶段。Curis公司多肽类的PD-L1小分子抑制剂CA-170(WO2015033299)已进入临床I期，BMS公开了多篇以苄基苯基醚类化合物为骨架的小分子PD-1/PD-L1抑制剂专利(WO2015034820，WO2017066227，WO2018009505)，广州再极也公开了一篇芳香乙烯类小分子PD-1/PD-L1抑制剂专利(WO2018006795)。目前小分子PD-1/PD-L1抑制剂在效果、安全性、以及可选择化合物等方面还远未能满足日益增加临床的需要。
相较于生物大分子，小分子化合物具有可口服、给药便利和明显更低的价格等多种优势，因此寻找小分子PD-1/PD-L1抑制剂具有重大的理论意义和应用价值。
发明内容
为了克服现有技术中的缺陷，本发明人进行了大量研究，通过大量筛选试验后意外发现，具有以下式I的化合物具有出乎意料地好的PD-1和/或PD-L1抑制作用：
本发明所述的式I化合物，可以选择性地抑制PD-1和/或PD-L1，可用于治疗和/或预防PD-1/PD-L1信号通路介导的疾病。这样的疾病包括但不限于癌症、自身免疫性疾病、感染性疾病和败血症。
基于上述发现，一方面，本发明提供了具有PD-1和/或PD-L1抑制活性的式I所示的联苯类化合物。本发明所述的式I化合物包括其可药用盐。
在另一方面，本发明提供了制备本发明所述式I化合物的方法。
在另一方面，本发明还提供了本发明式I化合物在制备用作PD-1和/或PD-L1抑制剂的药物中的应用。在又一方面，本发明还提供了本发明式I化合物在制备用于治疗或预防癌症、自身免疫性疾病、感染性疾病等疾病的药物中的应用。
在又一方面，本发明还提供了一种药物组合物，其包含本发明所述的式I化合物(包括其可药用盐)以及任选的可药用赋形剂或载体。
本发明所说的化合物，其化学结构如式I所示：
包括其可药用盐，
其中，
R
1选自氢、C
1-C
3烷基、CN、卤素、C
1-C
3卤代烷基；
R
2选自氢、C
1-C
3烷基、CN、卤素、C
1-C
3烷氧基；
L为任选取代的C
3-C
6烯基，所述C
3-C
6烯基中的一个CH
2任选地被-C(O)-、-S(O)-或-S(O)
2-所替代；
B为氢，或NR
6R
7，或任选取代的具有1到3个独立地选自氮、氧或硫的 杂原子的4元到7元饱和或部分不饱和杂环；其中
R
6和R
7独立地为氢、C
1-C
3烷基、C
1-C
3卤代烷基、羟基C
1-C
3烷基或羧基C
1-C
3烷基；
Z为氢、-CH
3、-CH＝CHAr或-OR
8；其中
R
8选自氢、C
3-C
6烯基、卤代C
1-C
4烷基、羟基C
1-C
4烷基、-(CH
2)
nX及-(CH
2)
nAr；
n为1、2、3或4；
X选自氢、-CH
3、-CF
3、C
1-C
4烷氧基、-N(CH
3)
2、任选经一个或两个卤素取代的C
3-C
6环烷基、-CN、-CO
2Rg、-C(O)NH
2、-C(O)N(CH
3)
2、
吗啉基、四氢吡喃基、任选经羟基取代的吡咯烷酮基及任选经一个或两个独立选自以下的基团取代的哌啶基：C
1-C
4烷基、羧基、羟基及C
1-C
4烷氧基羰基；
Rg选自氢及C
1-C
4烷基；及
Ar选自苯并二噁烷基、吲唑基、异喹啉基、异噁唑基、萘基、噁二唑基、苯基、吡啶基、嘧啶基及喹啉基；其中各环任选经1、2、3或4个独立选自以下的取代基取代：C
1-C
4烷氧基、C
1-C
4烷氧基羰基、C
1-C
4烷氧基羰基氨基、C
1-C
4烷基、C
1-C
4烷基羰基、C
1-C
4烷基磺酰基、甲酰氨基、甲酰氨基C
1-C
4烷基、-(CH
2)
qCO
2-(C
1-C
4烷基)、-(CH
2)
qOH、羧基、氰基、甲酰基、卤素、卤代C
1-C
4烷基、卤代C
1-C
4烷氧基、硝基、任选经一个氰基取代的苯基、任选经一个卤素取代的苯基氧基、苯基羰基、吡咯、四氢吡喃及
其中q为0、1、2、3或4且其中t、z及R
z如下定义：
t为0、1、2、3或4；
z为1、2或3；
各R
z独立选自C
1-C
4烷氧基、C
1-C
4烷氧基羰基、C
1-C
4烷氧基羰基C
1-C
4 烷基、C
1-C
4烷基、C
1-C
4烷基酰氨基、C
1-C
4烷基氨基、C
1-C
4烷基羰基、酰氨基、羧基、羧基C
1-C
4烷基、二(C
1-C
4烷基)酰氨基、二(C
1-C
4烷基)氨基、卤素、卤代C
1-C
4烷氧基、卤代C
1-C
4烷基、羟基、羟基C
1-C
4烷基、吗啉基、-NR
cR
d、(NR
cR
d)C
1-C
4烷基、-NR
eR
f、(NR
eR
f)C
1-C
4烷基、氧代、苯基及苯基C
1-C
4烷基，其中该苯基及该苯基C
1-C
4烷基的苯基部分任选经一个、两个或三个独立选自C
1-C
3烷基及卤素的基团取代；
R
c及R
d独立选自氢、C
2-C
4烯基羰基、C
1-C
4烷氧基羰基、C
1-C
6烷基、C
1-C
4烷基羰基、酰氨基C
1-C
4烷基、氨基C
1-C
4烷基、芳基C
1-C
4烷基、C
3-C
10环烷基、(C
3-C
10环烷基)C
1-C
4烷基、卤代C
1-C
4烷基羰基、杂环基C
1-C
4烷基、杂环基C
1-C
4烷基羰基、羟基C
1-C
6烷基及羟基C
1-C
4烷基羰基，其中该酰氨基C
1-C
4烷基、该氨基C
1-C
4烷基、该芳基C
1-C
4烷基、该(C
3-C
10环烷基)C
1-C
4烷基、该杂环基C
1-C
4烷基及该杂环基C
1-C
4烷基羰基的烷基部分任选经一个或两个独立选自羧基及羟基的基团取代；其中该羟基C
1-C
4烷基及该羟基C
1-C
4烷基羰基的烷基部分任选经一个或两个独立选自羧基及羟基的基团取代；且其中该芳基C
1-C
4烷基的芳基部分、该C
3-C
10环烷基、该(C
3-C
10环烷基)C
1-C
4烷基的环烷基部分及该杂环基C
1-C
4烷基及该杂环基C
1-C
4烷基羰基的杂环基部分各任选经一个、两个或三个独立选自C
1-C
4烷氧基羰基、C
1-C
4烷基及卤素的基团取代；
R
e及R
f与其所连接的原子一起形成选自吗啉及
的环；
R
3独立选自C
2-C
4烯基、C
1-C
4烷氧基、C
1-C
4烷基、氰基、卤素及卤代C
1-C
4烷基；
m为0、1或2；
R
4选自氢、C
1-C
4烷基、苄基、(C
3-C
6环烷基)C
1-C
3烷基、卤代C
1-C
4烷基、任选经第二个羟基取代的羟基C1-C6烷基及任选经氰基取代的吡啶基(C
1-C
3烷基)；及
R
5选自氢、C
1-C
4烷基、-(CH
2)
nN(CH
3)
2、羧基C
2-C
6烯基、羧基C
1-C
6烷基及羟基C
1-C
6烷基，其中该羧基C
1-C
6烷基及该羟基C
1-C
6烷基的烷基部分任选经一个羟基或苯基取代，其中该苯基进一步任选经羟基取代；或者，R
5选自以下基团：
其中
R
w为-CONH
2；
R
10选自氢、苄基及甲基；
各R
10’独立选自氢及C
1-C
3烷基；
R
11选自氢、C
1-C
3烷基及苄基；
R
12选自C
2-C
4烯基及C
1-C
4烷基；及
R
50选自氢、C
1-C
6烷基及C
1-C
6烷氧基羰基；
或
R
4及R
5与其所连接的氮原子一起形成选自以下的环：
s为0、1或2；
z为1、2或3；
p为0，1或2；
R
13及R
13’独立选自氢、羧基、羟基C
1-C
4烷基、氧代及-C(O)NHSO
2R
16；
R
14独立选自氢、羟基C
1-C
4烷基、氧代及羧基；
各R
15独立选自C
1-C
4烷氧基羰基、C
1-C
6烷基、羧基、卤素、羟基、羟基C
1-C
4烷基、-NR
c’R
d’及苯基氧基羰基，其中该苯基任选经硝基取代，其中R
c’及R
d’独立选自氢、C
1-C
4烷氧基羰基及C
1-C
4烷基羰基；及
R
16选自三氟甲基、环丙基、C
1-C
4烷基、二甲基氨基及经甲基取代的咪唑基。
对于上述式I所示化合物，优选地，
R
1和R
2独立地选自氢和C
1-C
3烷基；
L为-CH
2-CH＝CH-或-CH
2-CH
2-CH＝CH-，其中一个CH
2任选地被-C(O)-、-S(O)-或-S(O)
2-所替代，上述L定义中基团左侧与式I结构中B基团相连；
Z为甲基、-CH＝CHAr或-O(CH
2)
nAr；其中
n为1、2、3或4；
Ar选自苯并二噁烷基、吲唑基、异喹啉基、异噁唑基、萘基、噁二唑基、苯基、吡啶基、嘧啶基及喹啉基；其中各环任选经1、2、3或4个独立选自以下的取代基取代：C
1-C
4烷氧基、C
1-C
4烷氧基羰基、C
1-C
4烷氧基羰基氨基、C
1-C
4烷基、C
1-C
4烷基羰基、C
1-C
4烷基磺酰基、酰氨基、酰氨基C
1-C
4烷基、-(CH
2)
qCO
2C
1-C
4烷基、-(CH
2)
qOH、羧基、氰基、甲酰基、卤素、卤代C
1-C
4烷基、卤代C
1-C
4烷氧基、硝基、任选经一个氰基取代的苯基、任选经一个卤素取代的苯基氧基、苯基羰基、吡咯、四氢吡喃，其中q为0、1、2、3或4；
R
3独立选自C
1-C
4烷氧基和卤素。
对于上述式I所示化合物，较优选地，
R
1和R
2独立选自甲基及卤素；
Z为甲基、-CH＝CHAr或-OCH
2Ar；其中
Ar为任选经一个或两个独立选自以下的基团取代的吡啶基：C
1-C
4烷基、C
1-C
4烷基磺酰基、甲酰氨基、氰基及卤素；
R
3为卤素；且
m为1。
本发明所述的“卤素”为氟、氯、溴或碘。
本发明所述的“烷基”，是指具有1至约20个碳原子、通常为1至12个碳原子、优选为1至8个、1至6个甚至更优选为1至4个碳原子的直链或支链或环状的烷基。直链烷基的实例包括具有1至8个碳原子的那些，比如甲基、乙基、正丙基、正丁基、正戊基、正己基、正庚基和正辛基；支链烷基的实例包括但不限于异丙基、异丁基、仲丁基、叔丁基、新戊基、异戊基、异己基和2,2-二甲基丙基；环烷基的实例包括但不限于环丙基、环丁基、环戊基、环己 基、环庚基和环辛基。
本发明所述的“卤代烷基”包括单卤代烷基和多卤代烷基(其中所有卤原子可以相同或不同)。部分卤代的烷基是本文含义内的“卤代烷基”。卤代烷基的实例包括三氟甲基、1,1-二氯乙基、1,2-二氯乙基、1,3-二溴-3,3-二氟丙基等。
本发明所述的“烷氧基”，是指上述烷基与氧原子相连所形成的基团，其中，氧原子具有自由成键能力，如甲氧基、乙氧基、丙氧基、丁氧基、戊氧基、异丙氧基、特丁氧基、环丙氧基、环己基氧基等。
本发明所述的“药学上可接受的盐”或“可药用盐”是指药学上可接受的酸和碱加成盐和溶剂化物。这类药学上可接受的盐包括与酸形成的盐，所述酸包括盐酸、磷酸、氢溴酸、硫酸、亚磺酸、甲酸、对甲苯磺酸、甲磺酸、硝酸、苯甲酸、柠檬酸、酒石酸、马来酸、氢碘酸、链状羧酸如乙酸、HOOC-(CH
2)
n-COOH(n＝0-4)等；也包括与碱形成的盐，这些盐的阳离子包括钠、钾、钙、铵离子等。
本发明中，“取代的”指本文中所定义的有机基团(其中包含有与氢原子键合的一个或多个键)被一个或多个与非氢原子或原子团键合的键取代，所述非氢原子或原子团即为取代基。
本发明化合物的取代基包括例如卤素、烷基(优选C
1-8烷基、C
1-6烷基或C
1-4烷基)、烷氧基(优选C
1-8烷氧基、C
1-6烷氧基或C
1-4烷氧基)、卤代烷基(优选C
1-8卤代烷基、C
1-6卤代烷基或C
1-4卤代烷基)、卤代烷氧基(优选C
1-8卤代烷氧基、C
1-6卤代烷氧基或C
1-4卤代烷氧基)、羟基、羟基烷基(优选羟基C
1-8烷基、羟基C
1-6烷基或羟基C
1-4烷基)。
在另一方面，本发明提供了制备本发明式I化合物的方法：
包括使式A化合物
与式F化合物
在有机溶剂和还原剂的作用下经还原胺化形成式I化合物，其中R
1、R
2、R
3、R
4、R
5、L、B、Z和m如前面所定义。式F化合物可以为氨基酸或氨基醇或氨基酸的酯。当式F化合物为氨基酸的酯时，式A与式F化合物经还原胺化得到式I化合物的酯，经水解得到式I化合物。
上述还原胺化反应所用的还原剂可为硼氢化钠(NaBH
4)、氰基硼氢化钠(NaBH
3CN)及醋酸硼氢化钠(NaBH
3OAc)等；所用酸性催化剂可为醋酸、盐酸、三氟醋酸；所用的碱性催化剂可为TEA和DIPEA等；所用的干燥剂可为无水硫酸镁(MgSO
4)、无水硫酸钠(Na
2SO
4)及分子筛等；所用的溶剂可为CH
3OH、CH
2Cl
2及1,2-二氯乙烷等。
特别地，本发明还提供了所述式A化合物作为用于制备本发明所述式I化合物的中间体的用途。
式A化合物的制备
此外，本发明进一步提供了制备上述式A化合物的方法，包括两种方案：方案一包括以下步骤(a)-(c)：
(a)在碱的作用下，化合物1与化合物2进行亲核取代反应得到化合物3；
(b)在钯催化作用下，化合物3与化合物4进行铃木交叉偶联得到化合物5；
(c)化合物5在碱性条件下转变为化合物6，接下去与化合物7在碱性条件下发生酚的O-烃化反应得到化合物A；
上述亲核取代反应(a)以NaOH、Na2CO
3、K
2CO
3等碱为去酸剂，反应温度为25～140℃，反应时间为2～72小时。反应所用溶剂为常用溶剂，如水、甲醇、乙醇、乙腈、苯、二甲苯、丙酮、N,N'-二甲基甲酰胺、DMSO等。
上述铃木交叉偶联反应(b)以钯为催化剂，如Pd(PPh
3)
4、Pd(dffp)Cl
2、Pd(OAc)
2、Pd(dba)
3/PCy
3、PdCl
2、Pd(PPh
3)
4Cl
2等。或者以Ni为催化剂，如NiCl
2(dffp)、NiCl
2(dffp)/Zn、NiCl
2(dffp)/BuLi、NiCl
2(PPh
3)
2/PPh
3、NiCl
2(NEt
3)
2、NiCl
2(NEt
3)
2、NiCl(bipy)、Ni(TPPS)
3、Ni(COD)
2、NiCl
2(PPh
3)
2/n-BuLi、Ni{P(OMe)
3}
2Cl
2、NiCl
2(PCy
3)
2等。反应优选在碱存在下进行，所述碱例如为KOAc、K
3PO
4、K
2CO
3、NaOH、Ba(OH)
2、Na
2CO
3、CsF或NaHCO
3等。反应温度为25到140℃，反应时间为4～72小时。反应所用溶剂为常用溶剂，如乙醇、THF、异丙醇、DMSO、DMF、二氧六环、甲苯、水和DME等。
上述反应(c)第一步将羟基转变为好的离去基团，如-OMs、-OTs或溴， 以三乙胺、DIPEA有机碱为去酸剂，反应温度为0～90℃，反应时间为2～72小时。反应所用溶剂为常用溶剂，如水、甲醇、乙醇、异丙醇、四氢呋喃、二氯甲烷、丙酮等；第二步酚的O-烃化反应以NaOH、K
2CO
3、KOH等碱为去酸剂，反应温度为0～90℃，反应时间为2～72小时。反应所用溶剂为常用溶剂，如水、甲醇、乙醇、异丙醇、丙酮、DMF、DMSO、苯或二甲苯等；
式B化合物可由市场上直接购得。
方案二包括以下步骤(d)-(e)：
(d)在PPh
3和DIAD作用下，化合物4与化合物7通过Mitsunobu反应转化为化合物8；
(e)在钯催化作用下，化合物8与化合物3进行铃木交叉偶联得到化合物A；
上述Mitsunobu反应(d)可以以nBu
3P、PPh
3、DIAD或DEAD作为活化试剂，反应温度为-78℃到室温，反应时间为2～72小时。反应所用溶剂可以为常用低极性溶剂，如THF、乙醚、二氯甲烷、甲苯、乙酸乙酯、乙腈、DMF等。
上述铃木交叉偶联反应(e)可以以钯为催化剂，如Pd(PPh
3)
4、Pd(dffp)Cl
2、Pd(OAc)
2、Pd(dba)
3/PCy
3、PdCl
2、Pd(PPh
3)
4Cl
2等，或者以Ni为催化剂，如NiCl
2(dffp)、NiCl
2(dffp)/Zn、NiCl
2(dffp)/BuLi、NiCl
2(PPh
3)
2/PPh
3、NiCl
2(NEt
3)
2、NiCl
2(NEt
3)
2、NiCl(bipy)、Ni(TPPS)
3、Ni(COD)
2、NiCl
2(PPh
3)
2/n-BuLi、Ni{P(OMe)
3}
2Cl
2、NiCl
2(PCy
3)
2等。反应可以在碱存在下进行，所述碱例如KOAc、K
3PO
4、K
2CO
3、NaOH、Ba(OH)
2、Na
2CO
3、CsF或NaHCO
3等，反应 温度为25到140℃，反应时间为4～72小时。反应所用溶剂可以为常用溶剂，如乙醇、THF、异丙醇、DMSO、DMF、二氧六环、甲苯、水和DME等。
式I化合物可以采用常见的分离方法进行纯化，如萃取、重结晶、柱层析等。
按照上述说明方法，本发明制备了示例性的式I化合物(见表1)。
表1 本发明所述的代表性化合物
下面结合具体实例进一步阐述本发明内容，但本发明的保护范围并不仅仅局限于这些实例。本发明所述的百分比除特别注明外，均为重量百分比。说明书中所描述的数值范围，如计量单位、反应条件、化合物物理状态或百分比，均是为了提供明白无误的书面参考。本领域技术人员在实施本发明时，使用在此范围之外或有别于单个数值的温度、浓度、数量、碳原子数等，仍然有可能得到预期结果。
实施例中所使用的试剂缩写所代表的完整名称如下：
PdCl
2(dppf) 1,1'-双二苯基膦二茂铁二氯化钯
PdCl
2(dppf)CH
2Cl 1,1'-双二苯基膦二茂铁二氯化钯二氯甲烷络合物
DIAD 偶氮二甲酸二异丙酯
Ph
3P 三苯基膦
B
2(pin)
2 双联频哪醇硼酸酯
DIBAL-H 二异丁基氢化铝
TEA 三乙胺
MsCl 甲基磺酰氯
DMF N,N-二甲基甲酰胺
THF 四氢呋喃
EA 乙酸乙酯
PE 石油醚
DCM 二氯甲烷
实施例1
(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯的制备
将环丙基溴化镁(91mL,45.45mmol,0.5M/L in THF)在氮气保护0℃条件下缓慢滴加进3-溴-2-甲基苯甲醛(3g,15.15mmol)的THF溶液(70mL)中。在室温搅拌2h后，加入磷酸二乙酯(2.51g,18.18mmol)。混合物继续在室温下搅拌24h。然后用1N的HCl溶液(150mL)淬灭反应液。用乙酸乙酯(100mL)萃取三次，然后用无水Na
2SO
4干燥有机相。去除溶剂，残留物用硅胶柱(PE/EA＝100/1)纯化，得到浅黄色油状物(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(1.585g,5.21mmol,35％收率)。
1H NMR(400MHz,CDCl
3):δ7.45(d,J＝8.0Hz,1H),7.31(d,J＝8.0Hz,1H),7.00(t,J＝7.6Hz,1H),6.70(d,J＝15.6Hz,1H),5.98(dt,J＝15.6Hz,6.8Hz,1H),3.49(t,J＝6.8Hz,2H),2.83-2.76(m,2H),2.42(s,3H).
实施例2
(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-3,3-二氟吡咯烷的制备
将(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(1.48g,4.92mmol),3,3-二氟吡咯烷盐酸盐(1.41g,9.84mmol)和三乙胺(1.49g,14.76mmol)溶于1,4-二氧六环(20mL)，加热至100℃，反应18h。将反应液浓缩得到粗品，用硅胶柱层析纯化(PE/EA＝100/1to 20/1)，得到黄褐色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-3,3-二氟吡咯烷(810mg,2.46mmol,50％收率)。
1H NMR(400MHz,CDCl
3):δ7.42(d,J＝8.0Hz,1H),7.29(d,J＝7.6Hz,1H),6.98(t,J＝8.0Hz,1H),6.70(d,J＝15.6Hz,1H),6.01(dt,J＝15.2Hz,6.8Hz,1H),2.93(t,J＝13.6Hz,2H),2.76(t,J＝7.2Hz,2H),2.61(t,J＝7.2Hz,2H),2.45-2.37(m,5H),2.33-2.21(m,2H)；
MS Calcd:329；MS Found:329.9([M+H]
+).
实施例3
(E)-(3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲醇的制备
将(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-3,3-二氟吡咯烷(762mg,2.32mmol),(2-甲基-3-硼酸频那醇酯苯基)甲醇(660mg,2.67mmol)，PdCl
2(dppf)(189mg,0.23mmol)和K
2CO
3(640mg,4.64mmol)于氮气保护下溶于1,4-二氧六环(9mL)和水(3mL)的混合溶剂，80℃下搅拌4h。将反应液浓缩得到粗品，用硅胶柱层析纯化(PE/EA＝20/1to 10/1)，得到黄色油状物(E)-(3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲醇(1.05g,粗品)。
1H NMR(400MHz,CDCl
3):δ7.43-7.35(m,2H),7.25-7.15(m,2H),7.06(d,J＝6.8Hz,1H),6.98(d,J＝6.4Hz,1H),6.70(d,J＝15.6Hz,1H),6.10(dt,J＝15.6Hz,6.8Hz,1H),4.76(d,J＝2.8Hz,2H),2.95(t,J＝13.2Hz,2H),2.78(t,J＝6.8Hz,2H),2.63(t,J＝7.2Hz,2H),2.47-2.40(m,2H),2.33-2.22(m,2H),2.03(s,3H),1.99(s,3H)；
MS Calcd:371；MS Found:371.9([M+H]
+).
实施例4
(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
将甲磺酰氯(357mg,3.12mmol)在0℃下加入(E)-(3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲醇(580mg,1.56mmol)和三乙胺(473mg,4.68mmol)溶于10ml二氯甲烷的溶液中。反应液在室温下搅拌4h，然后加入50mL饱和氯化铵水溶液。该溶液用二氯甲烷(50mL)萃取三遍，合并的有机相用无水Na
2SO
4干燥，然后过滤。滤液减压浓缩得到粗品(E)-(3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲磺酸甲酯，无 需纯化，直接用于下一步。
(E)-(3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲磺酸甲酯(700mg,1.56mmol)，5-氯-4-羟基-2-甲基苯甲醛(240mg,1.42mmol)和K
2CO
3(392mg,2.84mmol)溶于DMF(5mL)的混合溶液在60℃下搅拌16h。加入50mL饱和氯化铵水溶液。该溶液用EA(50mL)萃取三遍，合并的有机相用无水Na
2SO
4干燥，然后过滤。滤液减压浓缩，硅胶柱层析纯化(PE/EA＝15/1to 10/1)得到黄色固体(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(480mg,0.91mmol,两步收率59％)。
1H NMR(400MHz,CDCl
3):δ10.10(s,1H),7.84(s,1H),7.49(d,J＝7.6Hz,1H),7.42(d,J＝7.2Hz,1H),7.30-7.26(m,1H),7.22-7.12(m,2H),7.02(d,J＝7.6Hz,1H),6.88(s,1H),6.72(d,J＝15.6Hz,1H),6.11(dt,J＝15.6Hz,6.8Hz,1H),5.23(s,2H),2.95(t,J＝13.2Hz,2H),2.78(t,J＝7.2Hz,2H),2.67-2.61(m,5H),2.48-2.41(m,2H),2.34-2.22(m,2H),2.07(s,3H),2.01(s,3H)；MS Calcd:323；MS Found:323.8([M+H]
+).
实施例5
(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
将(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(393mg,0.75mmol)和(S)-哌啶-2-羧酸(194mg,1.50mmol)溶于二氯甲烷(3mL)中于室温下搅拌4h。然后在0℃下加入甲醇(5mL)和氰基硼氢化钠(94mg,1.50mmol)。反应液在室温下搅拌16h。将1N盐酸水溶液缓慢加入反应液，调节反应液pH值到大约5，然后用二氯甲烷稀释(40mL)。混合液用水(40mL)洗一次，有机相用无水Na
2SO
4干燥，然后过滤。滤液减压浓缩得到残留物，用C18柱(with 10％-95％ACN in water)纯化得到白色固体(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(167.9mg,0.26mmol,35％收率)
1H NMR(400MHz,CDCl
3):δ7.49-7.38(m,3H),7.24-7.14(m,2H),7.08(d,J＝7.6Hz,1H),6.98(d,J＝7.2Hz,1H),6.76(s,1H),6.70(d,J＝16.0Hz,1H),6.10(dt,J＝15.2Hz,6.8Hz,1H),5.02(s,2H),4.36-4.20(m,1H),4.05-3.84(m,1H),3.74-3.59(m,1H),3.30-3.15(m,2H),2.94(t,J＝13.2Hz,2H),2.78(t,J＝6.8Hz,2H),2.63(t,J＝7.2Hz,2H),2.47-2.391(m,2H),2.36-2.22(m,5H),2.11-1.85(m,8H),1.80-1.56(m,3H),1.40-1.28(m,1H)；MS Calcd:636；MS Found:637.0([M+H]
+).
实施例6
(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶的制备
将(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(300mg,0.993mmol),哌啶(255mg,3.000mmol)溶于DMF(10mL)中，加入K
2CO
3(556mg,4.000mmol)，在室温下搅拌18h。反应用水淬灭，用乙酸乙酯萃取3次，合并有机相，用饱和食盐水洗3次，无水Na
2SO4干燥，浓缩。粗品用硅胶柱层析纯化(先用EA，再用DCM/MeOH＝10/1洗脱)得黄色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶(167mg,54％收率)。
LC-MS(m/z):308(M+1)。
实施例7
2-甲基-3-(羟甲基)苯硼酸频哪醇酯的制备
将3-溴-2-甲基苯甲醇(5.9g,29.500mmol),双联频哪醇硼酸酯(9.74g,38.346mmol),Pd(dppf)Cl
2CH
2Cl
2(1.68g,2.059mmol),KOAc(8.67g,88.469mmol)和1,4-二氧六环(70mL)的混合物在氮气保护下加热到80℃并搅拌5h。将反应液浓缩得到粗品，用硅胶柱层析纯化(PE/EA＝10/1to 5/1)，再用石油醚打浆洗1次得到白色固体2-甲基-3-(羟甲基)苯硼酸频哪醇酯(5.08g,20.484mmol,69％收率)。LC-MS(m/z):231(M-OH)。
实施例8
5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛的制备
将3-溴-2-甲基苯甲醇(10.450g,42.137mmol)，5-氯-4-羟基-2-甲基-苯甲醛(7.163g,42.137mmol)，PPh3(13.248g,50.564mmol)溶于超干四氢呋喃(120mL)中，在氮气保护下用冰/盐浴冷却至-7℃，缓慢滴加DIAD(10.214g,50.564mmol)。所得黄色溶液在室温下搅拌3h，TLC(PE/EA＝4/1)监控反应。反应用水淬灭，用乙酸乙酯萃取3次，合并有机相，浓缩，粗品用硅胶柱层析纯化(PE/EA＝10/1to 5/1)，再用石油醚打浆洗1次得到白色固体5-氯-2-甲基-4-[2-甲基-3-硼酸频那醇酯苄氧基]-苯甲醛(16.38g,40.950mmol,97％)。
LC-MS(m/z):401(M+1)。
实施例9
(E)-5-氯-4-((3’-(4-(哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
将(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶(167mg,0.5440mmol)，5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛(261mg,0.653mmol)，Pd(PPh
3)
4(94.3mg,0.0816mmol)，K
2CO
3(227mg,1.632mmol)，1,4-二氧六环(10.0mL)和H
2O(2.0mL)的混合液在氮气保护下加热至100℃并搅拌3h，TLC(EA)监控反应。将反应液浓缩得到粗品，用硅胶柱层析纯化(EA)得类白色固体(E)-5-氯-4-((3’-(4-(哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(262mg,0.523mmol,96％)。
LC-MS(m/z):502(M+1)。
实施例10
(S,E)-1-(5-氯-4-((2,2’-二甲基-3’-(4-(哌啶-1-基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
将(E)-5-氯-4-((3’-(4-(哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(262mg,0.523mmol)，L-哌啶-2-甲酸(236.1mg,1.831mmol)溶于DCM/MeOH(6mL/6mL)中，加入HOAc(10滴)，在室温下搅拌18h，加入NaBH3CN(197.7mg,3.138mmol)，继续搅拌2h。浓缩反应液。粗品先用硅胶柱(DCM/MeOH＝6/1,3/1)，再用制备薄层色谱版(DCM/MeOH＝4/1)纯化得白色固体(S,E)-1-(5-氯-4-((3’-(4-(哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(21.0mg,0.0342mmol,6.5％)。
1H NMR(400MHz,DMSO-d
6)δ7.51(d,J＝7.6Hz,1H),7.47(d,J＝7.6Hz,1H),7.33(s,1H),7.29(t,J＝7.6Hz,1H),7.24(t,J＝7.6Hz,1H),7.15(s,1H),7.07(d,J＝7.6Hz,1H),7.00(d,J＝7.6Hz,1H),6.82(d,J＝16.0Hz,1H),6.12(dt,J＝16.0,6.8Hz,1H),5.21(s,2H),3.75(d,J＝13.2Hz,1H),3.40(d,J＝13.2Hz,1H),3.15–2.92(m,4H),2.86–2.79(m,2H),2.59(d,J＝7.6Hz,2H),2.31(s,3H),2.29–2.19(m,1H),2.01(s,3H),1.97(s,3H),1.72–1.69(m,7H),1.47–1.39(m,7H).
LC-MS(m/z):615(M+1)。
实施例11
(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)吡咯烷的制备
黄色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)吡咯(172mg,0.587mmol,59％)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(300mg,0.993mmol)和四氢吡咯烷(255mg,3.000mmol)按照实施例6中的类似步骤制备而成。
LC-MS(m/z):294(M+1)。
实施例12
(E)-5-氯-4-((2,2’-二甲基-3’-(4-(吡咯-1-基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
浅黄色固体(E)-5-氯-4-((3’-(4-(吡咯-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(227mg,0.466mmol,79％)是由5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛(235mg,0.587mmol)按照实施例9中的类似步骤制备而成。
LC-MS(m/z):488(M+1)。
实施例13
(S,E)-1-(5-氯-4-((2,2’-二甲基-3’-(4-(吡咯-1-基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
(S,E)-1-(5-氯-4-((3’-(4-(吡咯-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(15.2mg,0.0253mmol,5.4％)是由(E)-5-氯-4-((3’-(4-(吡咯-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(227mg,0.466mmol)和L-哌啶-2-甲酸(210.4mg,1.631mmol)按照实施例10中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d6)δ7.50(dd,J＝13.5,7.7Hz,2H),7.33(s,1H),7.29(t,J＝7.5Hz,1H),7.23(t,J＝7.6Hz,1H),7.15(s,1H),7.07(d,J＝7.5Hz,1H),6.98(d,J＝7.4Hz,1H),6.77(d,J＝15.6Hz,1H),6.18(dt,J＝14.9,6.8Hz,1H),5.21(s,2H),3.75(d,J＝13.5Hz,1H),3.40(d,J＝13.4Hz,1H),3.11(t,J＝5.7Hz,1H),2.96(s,br,4H),2.87–2.79(m,1H),2.63-2.53(m,2H),2.32(s,3H),2.28-2.18(m,1H),2.01(s,3H),1.96(s,3H),1.91-1.80(m,4H),1.79-1.70(m,2H),1.57–1.33(m,4H),1.32-1.17(m,1H),0.90–0.79(m,1H).
LC-MS(m/z):601(M+1)。
实施例14
(E)-4-(3-溴-2-甲基苯基)-N,N-二甲基丁-3-烯-1-胺的制备
黄色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)二甲胺(670mg,2.509mmol,76％)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(1.0g,3.311mmol)和二甲胺盐酸盐(536.4mg,6.622mmol)按照实施例6中的类似步骤制备而成。
LC-MS(m/z):268(M+1)。
实施例15
(E)-5-氯-4-((3’-(4-(二甲胺)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
浅黄色固体(E)-5-氯-4-((3’-(4-(二甲胺-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(1.12g,2.430mmol,97％)是由(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)二甲胺(670mg,2.509mmol)和5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛(1.004g,2.509mmol)按照实施例9中的类似步骤制备而成。
LC-MS(m/z):462(M+1)。
实施例16
(S,E)-1-(5-氯-4-((3’-(4-(二甲胺)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
白色固体(S,E)-1-(5-氯-4-((3’-(4-(二甲胺-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(185.0mg,0.322mmol,13％yield)是由(E)-5-氯-4-((3’-(4-(二甲胺-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(1.12g,2.430mmol)和L-哌啶-2-甲酸 (1.097g,8.505mmol)按照实施例10中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d6)δ7.50(d,J＝7.6Hz,1H),7.44(d,J＝7.8Hz,1H),7.32(s,1H),7.27(t,J＝7.6Hz,1H),7.20(t,J＝7.6Hz,1H),7.14(s,1H),7.10–7.03(m,1H),6.99–6.92(m,1H),6.72(d,J＝15.7Hz,1H),6.15(dt,J＝15.6,6.8Hz,1H),5.20(s,2H),3.74(d,J＝13.4Hz,1H),3.09(t,J＝5.8Hz,1H),2.83(dd,J＝11.3,5.4Hz,1H),2.55–2.44(m,3H),2.44–2.34(m,2H),2.31(s,3H),2.28–2.13(m,6H),2.00(s,3H),1.95(s,3H),1.79–1.70(m,2H),1.58–1.29(m,5H).
LC-MS(m/z):575(M+1)。
实施例17
(E)-4-(3-溴-2-甲基苯基)-N,N-二乙基丁-3-烯-1-胺的制备
黄色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)二乙胺(685.0mg,2.322mmol,70％)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(1.0g,3.311mmol)和二乙胺(483.4mg,6.622mmol)按照实施例6中的类似步骤制备而成。
LC-MS(m/z):296(M+1)。
实施例18
(E)-5-氯-4-((3’-(4-(二乙胺)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
浅黄色固体(E)-5-氯-4-((3’-(4-(二乙胺-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(807mg,1.650mmol,71％)是由(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)二乙胺(685.0mg,2.322mmol)和5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛(928.8mg,2.322mmol)按照实施例9中的类似步骤制备而成。
LC-MS(m/z):490(M+1)。
实施例19
(S,E)-1-(5-氯-4-((3’-(4-(二乙胺)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
白色固体(S,E)-1-(5-氯-4-((3’-(4-(二乙胺-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(178.0mg,0.296mmol,36％yield)是由(E)-5-氯-4-((3’-(4-(二乙胺-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(400mg,0.818mmol)和L-哌啶-2-甲酸(369.3mg,2.863mmol)按照实施例10中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d
6)δ7.49(dd,J＝18.2,7.7Hz,2H),7.33(s,1H),7.29(t,J＝7.6Hz,1H),7.24(t,J＝7.6Hz,1H),7.15(s,1H),7.07(d,J＝7.5Hz,1H),6.99(d,J＝7.4Hz,1H),6.82(d,J＝15.7Hz,1H),6.15(dt,J＝14.9,6.9Hz,1H),5.21(s,2H),3.75(d,J＝13.4Hz,1H),3.40(d,J＝13.4Hz,1H),3.12(t,J＝5.9Hz,1H),2.87–2.78(m,1H),2.59-2.52(m,4H),2.31(s,3H),2.27–2.19(m,1H),2.01(s,3H),1.97(s,3H),1.82-1.67(m,3H),1.56-1.33(m,6H),1.24(s,1H),1.16(t,J＝7.2Hz,6H).
LC-MS(m/z):603(M+1)。
实施例20
(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)吗啉的制备
黄色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)吗啉(736mg,2.382mmol,72％)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(1.0g,3.311mmol)和吗啉(576.1mg,6.622mmol)按照实施例6中的类似步骤制备而成。
LC-MS(m/z):310(M+1)。
实施例21
(E)-5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
浅黄色固体(E)-5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(1.114g,2.215mmol,93％)是由(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)吗啉(736mg,2.382mmol)和5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛(952.8mg,2.382mmol)按照实施例9中的类似步骤制备而成。
LC-MS(m/z):504(M+1)。
实施例22
(S,E)-1-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
白色固体(S,E)-1-(5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(91.0mg,0.148mmol,6.7％yield)是由(E)-5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(1.114g,2.215mmol)和L-哌啶-2-甲酸(1.0g,7.752mmol)按照实施例10中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d
6)δ7.51(d,J＝7.6Hz,1H),7.43(d,J＝7.7Hz,1H),7.32(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.8Hz,1H),7.15(s,1H),7.07(d,J＝7.4Hz,1H),6.96(d,J＝7.6Hz,1H),6.72(d,J＝15.7Hz,1H),6.16(dt,J＝15.5,6.6Hz,1H),5.20(s,2H),3.88–3.81(m,1H),3.73(d,J＝13.4Hz,1H),3.58(t,J＝4.6Hz,3H),3.38(d,J＝13.4Hz,1H),3.10(t,J＝5.7Hz,1H),3.05(t,J＝4.8Hz,1H),2.86–2.78(m,1H),2.48–2.35(m,7H),2.31(s,3H),2.27–2.14(m,1H),2.01(s,3H),1.95(s,3H),1.81–1.66(m,3H),1.55–1.32(m,5H).
LC-MS(m/z):617(M+1)。
实施例23
(E)-((5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)氮亚烷基)二甲醇的制备
将(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(105.0mg,0.201mmol)，丝氨醇(64.0mg,0.703mmol)，二氯甲烷(10.0mL)，甲醇(2.0mL)和三乙胺(1.0mL)的混合溶液于室温下搅拌18h。然后加入NaBH
3CN(76.0mg,1.206mmol)，继续搅拌2h。用水淬灭反应，用二氯甲烷萃取2次，合并有机相，无水Na
2SO4干燥，浓缩。粗品用制备薄层色谱板纯化(DCM/MeOH＝12/1)得白色固体(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)丝氨醇(22.0mg,0.0368mmol,18％收率)。
1H NMR(400MHz,DMSO-d
6)δ7.50(d,J＝7.6Hz,1H),7.43(d,J＝7.6Hz,1H),7.39(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.15(s,1H),7.07(d,J＝7.6Hz,1H),6.96(d,J＝7.6Hz,1H),6.73(d,J＝15.6Hz,1H),6.15(dt,J＝15.6,6.8Hz,1H),5.20(s,2H),4.47(s,2H),3.69(s,2H),3.46–3.36(m,4H),2.91(t,J＝13.6Hz,2H),2.73(t,J＝6.8Hz,2H),2.59(t,J＝7.2Hz,2H),2.39(q,J＝7.2Hz,2H),2.30(s,3H),2.27–2.17(m,2H),2.01(s,3H),1.95(s,3H).
LC-MS(m/z):585(M+1)。
实施例24
(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸甲酯的制备
浅黄色固体(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸甲酯(166mg,0.278mmol, 69％yield)是由(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(210.0mg,0.402mmol)和甘氨酸甲酯盐酸盐(175mg,1.400mmol)按照实施例23中的类似步骤制备而成。
LC-MS(m/z):597(M+1)。
实施例25
(E)-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸的制备
将(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸甲酯(166mg,0.278mmol)溶于四氢呋喃(10.0mL)和水(2.0mL)中，加入LiOH
.H
2O(200mg,4.762mmol)，在室温下搅拌18h，TLC(EA)监控反应。浓缩反应液，加水(15.0mL)稀释，用1N稀盐酸调pH至4-5，过滤收集固体，烘干得白色固体(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸(85.0mg,0.146mmol,52％收率)。
1H NMR(400MHz,DMSO-d
6)δ7.63(s,1H),7.51(d,J＝7.6Hz,1H),7.48(d,J＝7.6Hz,1H),7.32–7.20(m,3H),7.08(d,J＝7.6Hz,1H),6.98(d,J＝7.6Hz,1H),6.78(d,J＝15.6Hz,1H),6.16(dt,J＝15.6,6.8Hz,1H),5.26(s,2H),4.11(s,2H),3.90(s,2H),3.46–3.22(m,4H),3.03(s,2H),2.59–2.54(m,2H),2.46–2.42(m,2H),2.41(s,3H),2.01(s,3H),1.96(s,3H).
LC-MS(m/z):583(M+1)。
实施例26
(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)L-丝氨酸苄酯的制备
浅黄色固体(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)L-丝氨酸苄酯(103mg,0.147mmol,70％yield)是由(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(110.0mg,0.210mmol)和L-丝氨酸苄酯盐酸盐(84.5mg,0.735mmol)按照实施例23中的类似步骤制备而成。
LC-MS(m/z):703(M+1)。
实施例27
(E)-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-丝氨酸的制备
白色固体(E)-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-丝氨酸(58mg,0.0948mmol,64％yield)是由(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)L-丝氨酸苄酯(103mg,0.147mmol)和LiOH H2O(200mg,4.762mmol)按照实施例25中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d
6)δ7.53-7.50(m,2H),7.43(d,J＝7.6Hz,1H),7.28(t,J＝7.6Hz,1H),7.23-7.16(m,2H),7.07(d,J＝7.6Hz,1H),6.96(d,J＝7.6Hz,1H),6.73(d,J＝15.6Hz,1H),6.15(dt,J＝15.6,6.8Hz,1H),5.22(s,2H),3.94–3.79(m,2H),3.74–3.62(m,2H),3.24(s,J＝5.6Hz,1H),2.91(t,J＝13.6Hz,2H),2.73(t,J＝7.2Hz,2H),2.59(t,J＝7.2Hz,2H),2.39(dd,J＝13.6,7.2Hz,2H),2.33(s,3H),2.28–2.17(m,2H),2.01(s,3H),1.95(s,3H).
LC-MS(m/z):613(M+1)。
实施例28
(5-氯-4-((3’-((E)-4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-苏氨酸苄酯的制备
浅黄色固体(5-氯-4-((3’-((E)-4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-苏氨酸苄酯(57.0mg,0.0796mmol,42％yield)是由(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(100.0mg,0.191mmol)和L-苏氨酸苄酯半草酸盐(149mg,0.668mmol)按照实施例23中的类似步骤制备而成。
LC-MS(m/z):717(M+1)。
实施例29
(5-氯-4-((3’-((E)-4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-苏氨酸的制备
白色固体(5-氯-4-((3’-((E)-4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-苏氨酸(43mg,0.0687mmol,86％yield)是由(5-氯-4-((3’-((E)-4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-苏氨酸苄酯(57.0mg,0.0796mmol)和LiOH H
2O(200mg,4.762mmol)按照实施例25中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d
6)δ7.51(dd,J＝7.6,1.6Hz,1H),7.45–7.40(m,2H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.18(s,1H),7.07(dd,J＝7.6,1.6Hz,1H),6.96(dd,J＝7.6,1.6Hz,1H),6.73(d,J＝15.6Hz,1H),6.15(dt,J＝15.6, 6.8Hz,1H),5.22(s,2H),3.93–3.80(m,2H),3.65(d,J＝13.6Hz,2H),2.97(s,1H),2.91(t,J＝13.6Hz,2H),2.73(t,J＝7.2Hz,2H),2.59(t,J＝7.2Hz,2H),2.39(q,J＝7.2Hz,2H),2.32(s,3H),2.29–2.15(m,2H),2.01(s,3H),1.95(s,3H),1.13(d,J＝6.4Hz,3H).
LC-MS(m/z):627(M+1)。
实施例30
(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)氮杂环丁烷-2-羧酸的制备
将(E)-5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(100.0mg,0.191mmol)，S-氮杂环丁烷-2-羧酸(67.5mg,0.668mmol)，DMF(2.0mL)，MeOH(2.0mL)，HOAc(3滴)的混合液在室温下搅拌18h，加入NaBH3CN(72.2mg,1.146mmol)，继续搅拌2h。浓缩反应液除掉甲醇，加水(15.0mL)稀释，过滤收集固体，烘干。粗品用制备薄层色谱板(DCM/MeOH＝4/1)纯化得白色固体(S,E)-1-(5-氯-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)氮杂环丁烷-2-羧酸(41.0mg,0.0675mmol,35％收率)。
1H NMR(400MHz,DMSO-d
6)δ7.50(d,J＝7.6Hz,1H),7.43(d,J＝7.6Hz,1H),7.35(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.17(s,1H),7.07(dd,J＝7.6,1.4Hz,1H),6.96(d,J＝7.6Hz,1H),6.73(d,J＝15.6Hz,1H),6.15(dt,J＝15.6,6.8Hz,1H),5.20(s,2H),3.91–3.81(m,2H),3.55(d,J＝13.2Hz,1H),3.21(s,1H),3.08(q,J＝8.0Hz,1H),2.91(t,J＝13.2Hz,2H),2.73(t,J＝7.2Hz,2H),2.59(t,J＝7.2Hz,2H),2.39(q,J＝7.2Hz,2H),2.34(s,3H),2.29–2.09(m,4H),2.01(s,3H),1.95(s,3H).
LC-MS(m/z):609(M+1)。
实施例31
(E)-4-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)硫代吗啉-1,1-二氧化物的制备
黄色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)硫代吗啉-1,1-二氧化物(815.0mg,2.283mmol,69％)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(1.0g,3.311mmol)和硫代吗啉-1,1-二氧化物(894.0mg,6.622mmol)按照实施例6中的类似步骤制备而成。
LC-MS(m/z):358(M+1)。
实施例32
(E)-5-氯-4-((3’-(4-(1,1-二氧化硫代吗啉)丁-1-烯-1基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
浅黄色固体(E)-5-氯-4-((3’-(4-(1,1-二氧化硫代吗啉)丁-1-烯-1基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(1.120g,2.033mmol,89％)是由(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)硫代吗啉-1,1二氧化物(815.0mg,2.283mmol)和5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛(913.2mg,2.283mmol)按照实施例9中的类似步骤制备而成。
LC-MS(m/z):552(M+1)。
实施例33
(S,E)-1-(5-氯-4-((3’-(4-(1,1-二氧化硫代吗啉)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
将(E)-5-氯-4-((3’-(4-(1,1-二氧化硫代吗啉)丁-1-烯-1基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(160.0mg,0.290mmol)，L-哌啶-2-甲酸 (131.0mg,1.016mmol)，DMF(2.0mL)，MeOH(2.0mL)，HOAc(3滴)的混合液在室温下搅拌18h，然后用冰/盐浴冷却至-5℃，加入NaBH3CN(109.6mg,1.740mmol)，再将反应液升至室温并继续搅拌2h。浓缩反应液除掉甲醇，加水(15.0mL)稀释，过滤收集固体，烘干。粗品用制备薄层色谱板(DCM/MeOH＝6/1)纯化得白色固体(S,E)-1-(5-氯-4-((3’-(4-(硫代吗啉-1,1二氧化物-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(32.2mg,0.0485mmol,16.7)。
1H NMR(400MHz,DMSO-d
6)δ7.51(d,J＝7.6Hz,1H),7.44(d,J＝7.6Hz,1H),7.32(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.15(s,1H),7.07(d,J＝7.6Hz,1H),6.96(d,J＝7.6Hz,1H),6.73(d,J＝15.6Hz,1H),6.16(dt,J＝15.6,6.8Hz,1H),5.20(s,2H),3.73(d,J＝13.6Hz,1H),3.38(d,J＝13.6Hz,1H),3.12–3.06(m,5H),2.96–2.94(m,3H),2.89–2.77(m,2H),2.66(t,J＝7.2Hz,2H),2.40(dd,J＝14.4,7.2Hz,2H),2.31(s,3H),2.23–1.96(m,1H),2.01(s,3H),1.95(s,3H),1.76–1.73(m,2H),1.48–1.38(m,4H).
LC-MS(m/z):665(M+1)。
实施例34
(R,E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶-3-醇的制备
白色固体(R,E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶-3-醇(475.0mg,1.471mmol,89％)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(500.0mg,1.656mmol)和(R)-3羟基哌啶盐酸盐(400.0mg,2.907mmol)按照实施例6中的类似步骤制备而成。
LC-MS(m/z):324(M+1)。
实施例35
(R,E)-5-氯-4-((3’-(4-(3-羟基哌啶-1-基)丁-1-烯-1基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
浅黄色固体(R,E)-5-氯-4-((3’-(4-(3-羟基哌啶-1-基)丁-1-烯-1基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(300mg,0.580mmol,39％)是由(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-(R)-3-羟基哌啶(475.0mg,1.471mmol)和5-氯-2-甲基-4-((2-甲基-3-硼酸频那醇酯苄基)氧)苯甲醛(588.4mg,1.471mmol)按照实施例9中的类似步骤制备而成。
LC-MS(m/z):518(M+1)。
实施例36
(S)-1-(5-氯-4-((3’-((E)-4-((R)-3-羟基哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
白色固体(S)-1-(5-氯-4-((3’-((E)-4-((R)-3-羟基哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(30.0mg,0.0476mmol,16.4％)是由(R,E)-5-氯-4-((3’-(4-(3-羟基哌啶-1-基)丁-1-烯-1基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(150mg,0.290mmol)和L-哌啶-2-甲酸(131.0mg,1.015mmol)按照实施例10中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d6)δ7.51(d,J＝7.6Hz,1H),7.47(d,J＝7.6Hz,1H),7.33(s,1H),7.29(t,J＝7.6Hz,1H),7.24(t,J＝7.6Hz,2H),7.15(s,1H),7.07(d,J＝7.6Hz,1H),7.00(d,J＝7.6Hz,1H),6.81(d,J＝15.6Hz,1H),6.11(dt,J＝15.6,6.8Hz,1H),5.21(s,2H),3.75(d,J＝13.5Hz,1H),3.41(d,J＝13.5Hz,1H),3.17–3.11(m,5H),2.87–2.79(m,2H),2.61(s,2H),2.31(s,3H),2.27–2.18(m,1H),2.01(s,3H),1.97(s,3H),1.81–1.69(m,4H),1.65-1.56(m,2H),1.54–1.34(m,5H).
LC-MS(m/z):631(M+1)。
实施例37
4-((3-溴-2-甲基苄基)氧基)-5-氯-2-羟基苯甲醛的制备
0℃下将Ph
3P(62.3g,237.8mmol)和DIAD(48.0g,237.8mmol)加入(3-溴-2-甲基苯基)甲醇(31.7g,158.5mmol)和5-氯-2,4-二羟基苯甲醛(30g,174.4mmol)的四氢呋喃溶液(200mL)中。反应液在氮气保护下室温搅拌过夜。然后将反应液浓缩，残余物用硅胶层析柱纯化(PE/EA＝30/1)。得到白色固体4-((3-溴-2-甲基苄基)氧基)-5-氯-2-羟基苯甲醛(12.6g,22％yield)。
1H NMR(400MHz,CDCl
3):δ11.42(s,1H),9.70(s,1H),7.58(d,J＝8.0Hz,1H),7.54(s,1H),7.41(d,J＝4Hz,1H),7.10(t,J＝6.8Hz,1H),6.57(s,1H),5.15(s,2H),2.44(s,3H).。
LC-MS(m/z):631.4(M+1)。
实施例38
5-((5-((3-溴-2-甲基苄基)氧基)-4-氯-2-甲酸基苯氧基)甲基)烟腈的制备
将甲磺酰氯(8.07g,70.8mmol)在0℃下加入5-(羟基甲基)烟腈(7.16g,53mmol)和三乙胺(10.7g,106mmol)的二氯甲烷溶液中。混合溶液在室温下搅拌4小时，然后加入300mL饱和氯化铵水溶液。将该溶液用二氯甲烷(200mL)萃取三遍，合并有机相，无水Na
2SO
4干燥并过滤。滤液减压浓缩得到粗品，无需纯化直接用于下一步。将该粗品、化合物4-((3-溴-2-甲基苄基)氧基)-5-氯-2-羟基苯甲醛(12.6g,35.4mmol)和碳酸钾(14.6g,105.9mmol)溶于DMF(100ml)中，在60℃下加热16个小时。然后加入50ml饱和氯化铵水溶液。该混合溶液用乙酸乙酯(300ml)萃取三次。合并有机相，无水Na
2SO
4干燥并过滤。滤液减压浓缩，得 到的粗品用硅胶层析柱纯化(PE/EA＝2/1)。得到白色固体5-((5-((3-溴-2-甲基苄基)氧基)-4-氯-2-甲酸基苯氧基)甲基)烟腈(7.5g,两步产率45％yield)。
1H NMR(400MHz,CDCl
3):δ10.27(s,1H),8.92-8.89(m,2H),8.08(s,1H),7.92(s,1H),7.60(d,J＝7.2Hz,1H),7.35(d,J＝7.6Hz,1H),7.10(t,J＝7.6Hz,1H),6.56(s,1H),5.30-5.13(m,4H),2.43(s,3H).
实施例39
(E)-3,3-二氟-1-(4-(2-甲基-3-(-4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)苯基)丁-3-烯-1-基)吡咯烷的制备
室温下在化合物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-3,3-二氟吡咯烷(800mg,2.43mmol)的1,4-二氧六烷(10ml)溶液中中加入B
2(pin)
2(679mg,2.67mmol),KOAc(715mg,7.29mmol)and PdCl
2(dppf)(178mg,0.24mmol)。该反应液在100℃下搅拌过夜。反应结束后将反应液浓缩，残留物用硅胶层析柱纯化(PE/EA＝50/1)。得到黄色油状物(E)-3,3-二氟-1-(4-(2-甲基-3-(-4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)苯基)丁-3-烯-1-基)吡咯烷(812mg,88％yield)。
MS Calcd:377.2；MS Found:378.2([M+H]
+).
实施例40
(E)-5-((4-氯-5-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)-2-甲酸基苯氧基)甲基)烟腈的制备
将化合物5-((5-((3-溴-2-甲基苄基)氧基)-4-氯-2-甲酸基苯氧基)甲基)烟腈(550mg,1.16mmol)和PdCl
2(dppf)(85mg,0.13mmol)在室温下加入化合物(E)-3,3-二氟-1-(4-(2-甲基-3-(-4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)苯基)丁-3-烯-1-基)吡咯烷(480mg，3.48mmol)的1,4-二氧六环(0.5ml)和K
2CO
3(480mg，3.48mmol)的水(0.5ml)混合溶液中。该反应液在80℃氮气保护下加热反应过夜。反应结束后将反应液浓缩，残留物用硅胶层析柱纯化(PE/EA＝10/1)。得到黄色固体(E)-5-((4-氯-5-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)-2-甲酸基苯氧基)甲基)烟腈(417mg,54％yield)。
MS Calcd:642.1；MS Found:643.2([M+H]
+).
实施例41
(S,E)-1-(5-氯-2-((5-氰基吡啶-3-基)甲氧基)-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)卞基)哌啶-2-羧酸的制备
将化合物(E)-5-((4-氯-5-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)-2-甲酸基苯氧基)甲基)烟腈(360mg,0.56mmol)和(S)-哌啶-2-羧酸(145mg,1.12mmol)溶于二氯甲烷(3mL)中在室温下搅拌4小时。然后在0℃下加入甲醇(3mL)和氰基硼氢化钠(72mg,1.12mmol)。该混合液在室温搅拌16小时。反应结束后缓慢加入1N盐酸溶液将pH值调至5，然后用二氯甲烷(40mL)稀释。混合溶液用水(40mL)洗一次。有机相用无水Na
2SO
4干燥并过滤。滤液减压浓缩，得到的粗品用C18柱(with 10％-95％ACN in water)分离纯化得到白色固体(S,E)-1-(5-氯-2-((5-氰基吡啶-3-基)甲氧基)-4-((3’-(4-(3,3-二氟吡咯烷-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)卞基)哌啶-2-羧酸(130mg,31％yield)。
1H NMR(400MHz,CD
3OD):δ8.96(d,J＝11.2Hz,1H),8.41(s,1H),7.59(s,1H),7.50-7.47(m,2H),7.29-7.21(m,2H),7.11(s,1H),7.09(s,1H),7.25(d,J＝7.2Hz,1H),6.96(d,J＝15.6Hz,1H),6.12(t,J＝8.0Hz,1H),5.39(s,2H),5.33(s,2H),4.48-4.43(m,2H),3.97(t,J＝11.6Hz,3H),3.78(t,J＝7.2Hz,2H),3.51-3.43(m,3H),3.01(s,1H),2.76-2.66(m,4H),2.30(s,1H),2.08(s,3H),2.03(s,3H),1.85(s,3H),1.60-1.61(m,2H)；
MS Calcd:754；MS Found:755.4([M+H]
+).
实施例42
(E)-3-(3-溴-2-甲基苯基)丙烯酸乙酯的制备
0℃下将2-(二乙氧基磷酰基)乙酸乙酯(4.98g,22.2mmol)加入NaH(888mg,60％，22.2mmol)的DMF(50mL)溶液中。反应液在室温搅拌2小时。然后加入3-溴-2-甲基苯甲醛(4.0g,20.2mmol)。反应液继续在室温搅拌过夜。加水终止反应，用石油醚(50ml)萃取两次。无水Na
2SO
4干燥并过滤。滤液减压浓缩，得到的黄色油状物粗品(E)-3-(3-溴-2-甲基苯基)丙烯酸乙酯(5.83g,粗品)。粗品不经纯化直接用于下一步。
MS Calcd:268；MS Found:269.0([M+H]
+)
实施例43
(E)-3-(3-溴-2-甲基苯基)丙烯酸的制备
室温下将LiOH.H
2O(1.87g,44.61mmol)缓慢加入(E)-3-(3-溴-2-甲基苯基)丙烯酸乙酯(4.0g,14.87mmol)在乙醇(40mL)和水(10ml)的混合溶液中。反应液在氮气保护下室温搅拌6小时。反应结束后将反应液减压蒸馏除去溶剂，残留物溶于水(30mL)，用3N盐酸将溶液pH值调节至5-6，过滤水洗得到白色固体(E)-3-(3-溴-2-甲基苯基)丙烯酸(3.30g,93％yield)。
1H NMR(400MHz,CDCl
3):δ8.09(d,J＝16.0Hz,1H),7.60(d,J＝9.2Hz,1H),7.48(d,J＝3.6Hz,1H),7.09(t,J＝8.0Hz,1H),6.33(d,J＝15.6Hz,1H),2.52(s,3H).
实施例44
(E)-3-(3-溴-2-甲基苯基)-1-(3,3-二氟吡咯烷-1-基)丙-2-烯-1-酮的制备
室温下向(E)-3-(3-溴-2-甲基苯基)丙烯酸(2.0g,8.3mmol)和3,3-二氟吡咯烷(2.38g,16.6mmol)的二氯甲烷溶液(40mL)中加入HATU(4.73g,12.45mmol)和DIAD(2.14g,16.6mmol)。反应液在氮气保护下室温搅拌过夜。反应结束后将反应液浓缩，残留物通过硅胶柱层析分离得到黄色油状物(2.18g,75％yield)。MS Found:332.0([M+H]
+).
实施例45
(E)-1-(3,3-二氟吡咯烷-1-基)-3-(2-甲基-3-(-4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)苯基)丙-2-烯-1-酮的制备
室温下在(E)-3-(3-溴-2-甲基苯基)-1-(3,3-二氟吡咯烷-1-基)丙-2-烯-1-酮(1.62g,4.9mmol)和B
2(Pin)
2(1.36g,5.39mmol)的1,4-二氧六烷(20ml)溶液中中加入KOAc(1.44g,14.7mmol)和PdCl
2(dppf)(359mg,0.49mmol)。该反应液在氮气保护下80℃下搅拌过夜。反应结束后将反应液浓缩，残留物用硅胶层析柱纯化(PE/EA＝30/1)。得到黄色油状物(E)-1-(3,3-二氟吡咯烷-1-基)-3-(2-甲基-3-(-4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)苯基)丙-2-烯-1-酮(1.7g,92％yield)。
MS Calcd:377.2；MS Found:378.2([M+H]
+)
实施例46
(E)-5-((4-氯-5-((3’-(3-(3,3-二氟吡咯烷-1-基)-3-氧丙-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)-2-甲酸基苯氧基)甲基)烟腈的制备
将K
2CO
3(600mg,4.35mmol)的水溶液(1ml)和PdCl
2(dppf)(106mg,0.15mmol)在室温下加入化合物(E)-1-(3,3-二氟吡咯烷-1-基)-3-(2-甲基-3-(-4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)苯基)丙-2-烯-1-酮(600mg,1.59mmol)和化合物5-((5-((3-溴-2-甲基苄基)氧基)-4-氯-2-甲酸基苯氧基)甲基)烟腈(684mg,1.45mmol)的1,4-二氧六环溶液(8ml)中。该反应液在氮气保护下80℃加热反应过夜。反应结束后将反应液浓缩，残留物用硅胶层析柱纯化(PE/EA＝2/1)。得到黄色固体(E)-5-((4-氯-5-((3’-(3-(3,3-二氟吡咯烷-1-基)-3-氧丙-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)-2-甲酸基苯氧基)甲基)烟腈(430mg,46％yield)。
MS Calcd:641；MS Found:642.2([M+H]
+).
实施例47
(S,E)-1-(5-氯-2-((5-氰基吡啶-3-基)甲氧基)-4-((3’-(4-(3,3-二氟吡咯烷-1-基)-3-氧丙-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)卞基)哌啶-2-羧酸的制备5193
将化合物(E)-5-((4-氯-5-((3’-(3-(3,3-二氟吡咯烷-1-基)-3-氧丙-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)-2-甲酸基苯氧基)甲基)烟腈(430mg,0.67mmol)和(S)-哌啶-2-羧酸(173mg,1.34mmol)溶于二氯甲烷(3mL)中在室温下搅拌4小时。然后在0℃下加入氰基硼氢化钠(50mg,0.8mmol)。该混合液在室温搅拌16小时。反应结束后缓慢加入1N盐酸溶液将pH值调至5，然后用二氯甲烷(40mL)稀释。混合溶液用水(40mL)洗一次。有机相用无水Na
2SO
4干燥并过滤。滤液减压浓缩，得到的粗品用C18柱(with 10％-95％ACN in water)分离纯化得到白色固体(S,E)-1-(5-氯-2-((5-氰基吡啶-3-基)甲氧基)-4-((3’-(4-(3,3-二氟吡咯烷-1-基)-3-氧丙-1-烯-1-基)-2,2’-二甲基-[1,1’-联苯]-3-基)甲氧基)卞基)哌啶-2-羧酸(110mg,21％yield)。
1H NMR(400MHz,CD
3OD):δ8.85-8.23(m,2H),8.29(s,1H),7.95(d,J＝15.6Hz,1H),7.63(d,J＝7.6Hz,1H),7.54(s,1H),7.38(d,J＝7.2Hz,1H),7.22-7.15(m,2H),7.06-7.01(m,2H),6.74(dd,J＝15.2Hz,J＝39.2Hz,1H),5.26(s,2H),5.20(s,2H),4.31(d,J＝13.2Hz,1H),4.18-4.03(m,2H),3.91(t,J＝6.8Hz,1H),3.79(t,J＝13.2Hz,1H),3.69(t,J＝7.2Hz,1H),3.38(s,1H),2.75(s,1H),2.47-2.36(m,2H),2.00-1.83(m,7H),1.68-1.40(m,6H)；
MS Calcd:754；MS Found:755.4([M+H]
+).
实施例48
(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-(S)-3-羟基吡咯烷的制备
无色油状物(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-(S)-3-羟基吡咯烷(240mg,0.77mmol,47.1％)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(500mg,1.64mmol)和(S)-3-羟基吡咯烷(400mg,4.60mmol)按照实施例14中的类似步骤制备而成。
LC-MS(m/z):311(M+1)。
实施例49
(S，E)-5-溴-4-((3'-(4-(3-羟基吡咯烷-1-基)丁-1-烯-1-基)-2,2'-二甲基-[1，1,1'-联苯]-3-基)甲氧基)-2-甲基苯甲醛的制备
灰白色固体(S，E)-5-氯-4-((3'-(4-(3-羟基吡咯烷-1-基)丁-1-烯-1-基)-2,2'-二甲基-[1，1,1'-联苯]-3-基)甲氧基)-2-甲基苯甲醛(140mg,0.27mmol,35.9％)是由(E)-1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-(S)-3-羟基吡咯烷(240mg,0.77mmol,)和5-氯-2-甲基-4-[2-甲基-3-(4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)-苄氧基]-苯甲醛(310mg,0.77mmol)按照实施例17中的类似步骤制备而成。
LC-MS(m/z):505(M+1)。
实施例50
(S)-1-(5-氯-4-((3'-((E)-4-((S)-3-羟基吡咯烷-1-基)丁-1-烯-1-基)-2-，2'-二甲基-[1,1'-联苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
白色固体(S)-1-(5-氯-4-((3'-((E)-4-((S)-3-羟基吡咯烷-1-基)丁-1-烯-1-基)-2-，2'-二甲基-[1,1'-联苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(18mg,0.029mmol,14.7％yield)是由(S，E)-5-氯-4-((3'-(4-(3-羟基吡咯烷-1-基)丁-1-烯-1-基)-2,2'-二甲基-[1，1,1'-联苯]-3-基)甲氧基)-2-甲基苯甲醛(100mg,0.20mmol)和L-哌啶-2-甲酸(89mg,0.69mmol)按照实施例18中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d6)δ9.89(s,br,1H),7.50(dd,J＝13.3,7.7Hz,2H),7.33(s,1H),7.27(dt,J＝16.1,7.6Hz,2H),7.15(s,1H),7.07(d,J＝7.6Hz,1H),7.00(d,J＝7.4Hz,1H),6.83(d,J＝15.6Hz,1H),6.13(dt,J＝15.0,6.9Hz,1H),5.45(s,1H),5.21(s,2H),4.44(s,1H),3.77(d,J＝13.4Hz,1H),3.42(d,J＝13.4Hz,1H),3.36-3.07(m,5H),2.88–2.79(m,1H),2.62(q,J＝7.4Hz,2H),2.32(s,3H),2.29–2.22(m,1H),2.18-2.13(m,1H),2.01(s,3H),1.97(s,3H),1.93–1.82(m,1H),1.78-1.73(m,2H),1.59-1.15(m,5H).
LC-MS(m/z):618(M+1)。
实施例51
叔丁基-N-[2-(丙基-2-烯胺)乙基]氨基甲酸酯的制备
将N-叔丁氧基羰基-1,2-乙二胺(2.1g,13.125mmol)和TEA(3.977g,39.375mmol)溶于THF(30mL)中，用冰/盐浴冷却至0℃，滴加丙烯酰氯(2.376g,26.250mmol,2.0eq)，所得混合液在室温下搅拌2h。将生成的固体过滤掉，浓缩滤液得黄色油状叔丁基-N-[2-(丙基-2-烯胺)乙基]氨基甲酸酯粗产物(3.12g，13.125mmol,产率按100％计)，不经纯化直接用于下一步。
LC-MS(m/z):215(M+1)。
实施例52
N-(2-氨基乙基)丙烯酰胺的制备
将黄色油状叔丁基-N-[2-(丙基-2-烯胺)乙基]氨基甲酸酯粗产物(3.12g,13.125mmol)溶于DCM(30mL)中，加入TFA(30mL)，在室温下搅拌1h。浓缩得N-(2-氨基乙基)丙烯酰胺(4.17g，13.125mmol,产率按100％计)粗产物，不经纯化直接用于下一步。
LC-MS(m/z):115(M+1)。
实施例53
(E)-N-(2-{5-氯-4-[2,2’-二甲基-3’-(4-吗啉-4-基-丁基-1-烯基)-二苯基-3-基甲氧基]-2-甲基-苯并氨基}-乙基)-丙烯酰胺的制备
将(E)-5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(300mg,0.596mmol)，N-(2-氨基乙基)丙烯酰胺(1.5g粗品,4.721mmol)溶于DCM/MeOH(20mL/2mL)中，加入TEA(2mL)，在室温下搅拌18h，加入NaBH3CN(227.1mg,3.576mmol)，继续搅拌2h。反应用水淬灭，用乙酸乙酯萃取3次，合并有机相，无水Na
2SO
4干燥，浓缩。粗品用制备薄层色谱版(DCM/MeOH＝6/1)纯化一次，再用制备薄层色谱版(EA)纯化一次得黄色固体(E)-N-(2-{5-氯-4-[2,2’-二甲基-3’-(4-吗啉-4-基-丁基-1-烯基)-二苯基-3-基甲氧基]-2-甲基-苯并氨基}-乙基)-丙烯酰胺(37.0mg,0.0616mmol,10.3％yield)。
1H NMR(400MHz,DMSO-d
6)δ7.50(dd,J＝7.9,4.7Hz,2H),7.43(dd,J＝7.8,3.3Hz,2H),7.40–7.09(m,6H),7.07(dp,J＝12.0,4.3,3.6Hz,2H),6.96(dd,J＝7.6,4.8Hz,2H),6.72(dd,J＝15.7,4.6Hz,2H),6.29–6.01(m,4H),5.56(td,J＝10.2,2.3Hz,1H),5.23-5.13(m,4H),3.66–3.54(m,6H),3.46(d,J＝7.5Hz,1H), 3.31–3.09(m,3H),2.64(tq,J＝19.9,7.0Hz,3H),2.42(dd,J＝15.0,8.5Hz,19H),2.31(s,3H),2.01(s,3H),1.95(s,3H).
LC-MS(m/z):602.3(M+1)。
实施例54
3-(3-溴-2-甲基-苯基)-丙烯酸乙酯的制备
将NaH(60％,484mg,12.120mmol)悬浮于超干THF(30mL)中，用冰/盐浴冷却至0℃，加入膦酰基乙酸三乙酯(2.49g,11.116mmol)。在此温度下搅拌30min后滴加3-溴-2-甲基苯甲醛(2.0g,10.102mmol)的THF(5mL)溶液，并继续搅拌1h。反应用饱和NH
4Cl溶液淬灭，用乙酸乙酯萃取2次。合并有机相，饱和食盐水洗1次，无水Na
2SO
4干燥，浓缩得黄色油状3-(3-溴-2-甲基-苯基)-丙烯酸乙酯粗品(3.7g,10.102mmol,产率按100％计)，不经纯化直接用于下一步。
LC-MS(m/z):269(M+1)。
实施例55
3-(3-溴-2-甲基-苯基)-丙烯-2-1-醇的制备
将3-(3-溴-2-甲基-苯基)-丙烯酸乙酯(3.7g,10.102mmol,1.0eq)溶于超干DCM(30mL)中，氮气球置换3次氮气，用冰/盐浴冷却至0℃，滴加DIBAL-H(1mol/L的甲苯溶液，21.2mL,21.2 00mmol,2.1eq)，并继续在此温度下搅拌1h。反应用冰/水淬灭，用1N HCl(15mL)酸化。过滤除去固体，滤液用DCM萃取3次。合并有机相，浓缩。粗品用硅胶柱纯化(PE/EA＝4/1洗脱)得无色油状3-(3-溴-2-甲基-苯基)-丙烯-2-1-醇(2.1g,9.292mmol,92.0％yield)。
LC-MS(m/z):227(M+1)。
实施例56
甲磺酸3-(3-溴-2-甲基-苯基)-烯丙基酯的制备
将3-(3-溴-2-甲基-苯基)-丙烯-2-1-醇(452mg,2.000mmol,)溶于超干DCM(10mL)中，用冰/盐浴冷却至0℃，加入TEA(606mg,6.000mmol)，滴加MsCl(456mg,4.000mmol)。所得混合液在室温下搅拌4h。反应用DCM稀释，用水洗3次。有机相用无水Na
2SO
4干燥，浓缩得黄色油状甲磺酸3-(3-溴-2-甲基-苯基)-烯丙基酯粗品(626mg,2.000mmol,产率按100％计)，不经纯化直接用于下一步。
LC-MS(m/z):305(M+1)。
实施例57
4-[3-(3-溴-2-甲基-苯基)-烯基]吗啉的制备
将甲磺酸3-(3-溴-2-甲基-苯基)-烯丙基酯(626mg粗品,2.000mmol)溶于DMF(10mL)中，加入吗啉(348mg,4.000mmol)和K
2CO
3(834mg,6.000mmol)，在室温下搅拌18h。反应用水淬灭，用乙酸乙酯萃取3次。合并有机相，用饱和食盐水洗3次，浓缩。粗品用硅胶柱纯化(先用EA洗脱，再用DCM/MeOH＝6/1)得4-[3-(3-溴-2-甲基-苯基)-烯基]吗啉(530mg,1.797mmol,89.8％yield)。
LC-MS(m/z):296(M+1)。
实施例58
5-氯-2-羟基-4-[2-甲基-3-(4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)-苄氧基]-苯甲醛的制备
浅黄色固体5-氯-2-羟基-4-[2-甲基-3-(4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷 -2-基)-苄氧基]-苯甲醛(1.33g,3.308mmol,33％yield)是由2-甲基-3-(羟甲基)苯硼酸频哪醇酯(实施例15,2.48g,10.000mmol)和5-氯-2,4-二羟基-苯甲醛(1.72g,10.000mmol)按照实施例16中的类似步骤制备而成。
LC-MS(m/z):403(M+1)。
实施例59
5-氯-4-((3’-(4-(哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-羟基苯甲醛的制备
橙色固体5-氯-4-((3’-(4-(哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-羟基苯甲醛(422mg,0.859mmol,84.5％yield)是由5-氯-2-羟基-4-[2-甲基-3-(4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)-苄氧基]-苯甲醛(409mg,1.017mmol)和4-[3-(3-溴-2-甲基-苯基)-烯基]吗啉(300mg,1.017mmol)按照实施例17中的类似步骤制备而成。
LC-MS(m/z):492(M+1)。
实施例60
5-(氯甲基)烟腈的制备
将(5-氰基吡啶-3-基)-甲醇(288mg,2.149mmol)溶于DCM(10mL)中，加入SOCl
2(3.0mL)，在室温下搅拌2h。浓缩得白色固体5-(氯甲基)烟腈粗品(422mg,产率按100％计)，不经纯化直接用于下一步。
LC-MS(m/z):153(M+1)。
实施例61
5-{4-氯-5-[2,2’-二甲基-3’-(3-吗啉-4-基-丙基)-双苯-3-甲氧基]-2-甲基-苯氧甲基}-烟腈的制备
将5-(氯甲基)烟腈(422mg粗品,2.149mmol)溶于DMF(10mL)中，加入5-氯-4-((3’-(4-(哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-羟基苯甲醛(422mg,0.859mmol)和K
2CO
3(597mg,4.30mmol)，在室温下搅拌18h。反应用水淬灭，用乙酸乙酯萃取3次。合并有机相，用饱和食盐水洗3次，无水Na
2SO
4干燥，浓缩。粗品用硅胶柱纯化(DCM/MeOH＝10/1洗脱)得橙色固体5-{4-氯-5-[2,2’-二甲基-3’-(3-吗啉-4-基-丙基)-双苯-3-甲氧基]-2-甲基-苯氧甲基}-烟腈(503mg,0.829mmol,96.5％yield)。
LC-MS(m/z):608(M+1)。
实施例62
(S)-1-{5-氯-2-(5-氰基吡啶-3-甲氧基)-4-[2,2’-二甲基-3’-(3-吗啉-4-基-丙基)-二苯基-3-甲氧基]-苯基}-哌啶-2-羧酸的制备
浅黄色固体(S)-1-{5-氯-2-(5-氰基吡啶-3-甲氧基)-4-[2,2’-二甲基-3’-(3-吗啉-4-基-丙基)-二苯基-3-甲氧基]-苯基}-哌啶-2-羧酸(10.4mg,0.0144mmol,3.2％yield)是由5-{4-氯-5-[2,2’-二甲基-3’-(3-吗啉-4-基-丙基)-双苯-3-甲氧基]-2-甲基-苯氧甲基}-烟腈(260mg,0.428mmol)和L-哌啶-2-甲酸(221mg,0.713mmol)按照实施例18中的类似步骤制备而成
1H NMR(400MHz,CD
3OD):δ9.01(dd,J＝5.6,2.0Hz,2H),8.46(s,1H),7.50(d,J＝7.7Hz,2H),7.42(s,1H),7.25(dt,J＝19.0,7.5Hz,2H),7.12(s,1H),7.08(d,J＝7.5Hz,1H),7.00(d,J＝7.4Hz,1H),6.84(d,J＝15.7Hz,1H),6.17(dt,J＝15.6,6.6Hz,1H),5.39(s,2H),5.33(s,2H),5.27(s,2H),3.78(d,J＝13.7Hz,1H),3.63(s,1H),3.13(d,J＝6.5Hz,3H),2.88(d,J＝8.5Hz,1H),2.42(t,J＝4.5Hz,4H),2.28(m,1H),2.07-1.87(m,7H),1.82-1.77(m,2H),1.76-1.71(m,1H),1.51-1.46(m,4H),1.43-1.31(m,1H).
LC-MS(m/z):721.3(M+1)。
实施例63
(E)-2-((5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)氨基)丙烷-1,3-二醇的制备
白色固体(E)-2-((5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)氨基)丙烷-1,3-二醇(71mg，0.12mol，65.0％yield)由5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(95mg，0.19mmol)和2-氨基-1,3-丙二醇盐酸盐(127.5mg，1.0mmol)按照实施例6的类似步骤制备而成。
1H-NMR(400MHz,DMSO-d
6)δppm 7.50(d,J＝7.7Hz,1H),7.43(d,J＝8.0Hz,1H),7.38(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.14(s,1H),7.07(d,J＝7.3Hz,1H),6.96(d,J＝7.4Hz,1H),6.72(d,J＝15.7Hz,1H),6.15(dt,J＝15.5,6.5Hz,1H)，5.20(s,2H),4.44(s,2H),3.67(s,2H),3.58(t,J＝4.6Hz,4H),3.47-3.35(m,4H),2.56(p,J＝5.7Hz,1H),2.48-2.35(m,9H),2.29(s,3H),2.01(s,3H),1.95(s,3H)。
MS Calcd:578.3MS Found:579.4([M+H]
+).
实施例64
(E)-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸甲酯的制备
浅黄色固体(E)-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸甲酯(55.0mg,0.095mmol,55.0％yield)由5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(88mg，0.17mmol)和甘氨酸甲酯盐酸盐(90.0mg,0.7mmol)按照实施例6的类似步骤制备而成。
MS Calcd:576.2MS Found:577.4([M+H]
+).
实施例65
(E)-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸
(E)-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸(12.5mg,0.022mmol,23.4％yield)由(E)-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)甘氨酸甲酯(55.0mg，0.095mol)在LiOH/THF条件下按照实施例25中的类似步骤制备而成。
1H-NMR(400MHz,DMSO-d
6)δppm 7.54-7.40(m,3H),7.28(t,J＝7.6Hz,1H,),7.25-7.16(m,2H),7.07(d,J＝7.5Hz,1H),6.96(d,J＝7.5Hz,1H),6.72(d,J＝15.6Hz,1H),6.15(dt,J＝15.3,6.4Hz,1H),5.23(s,2H),3.84(s,2H),3.57(t,J＝4.6Hz,4H),3.12(s,2H),2.48-2.36(m,8H),2.34(s,3H),2.01(s,3H),1.95(s,3H)
MS Calcd:562.2MS Found:563.3([M+H]
+).
实施例66
(E)-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-脯氨酸的制备
类白色固体(E)-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)-L-脯氨酸(11.0mg,0.018mmol,9.0％yield)由5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(100mg，0.20mmol)和L-脯氨酸盐酸盐(151mg,1.0mmol)按照实施例18中的类似步骤制备而成。
1H-NMR(400MHz,DMSO-d
6)δppm 7.51(d,J＝7.6Hz,1H),7.43(d,J＝7.7Hz,1H),7.39(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.16(s,1H),7.07(d,J＝7.5Hz,1H),6.96(d,J＝7.4Hz,1H),6.72(d,J＝15.6Hz,1H),6.16(dt,J＝15.6,6.4Hz,1H),5.21(s,2H),3.97(d,J＝13.1Hz,1H),3.58(s,2H),3.61-3.49(m,5H)3.23(dd,J＝8.9,5.9Hz,1H),3.01-2.88(m,1H),2.48-2.37(m,8H),2.35(s,3H),2.17-2.03(m,1H),2.01(s,3H),1.95(s,3H),1.91-1.80(m,1H),1.80-1.64(m,2H).
MS Calcd:602.3MS Found:603.4([M+H]
+).
实施例67
(E)-4-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-2,6-二甲基吗啉的制备
中间体(E)-4-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-2,6-二甲基吗啉(420mg，1.24mmol，62.3％yield)由2,6-二甲基吗啉(345mg，3.0mmol)和(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(604.0mg，2.0mmol)按照实施例14中的类似步骤制备而成。
MS Calcd:337.1，339.1MS Found:338.0，340.2([M+H]
+).
实施例68
(E)-5-氯-4-((3’-(4-(2,6-二甲基吗啉)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
中间体(E)-5-氯-4-((3’-(4-(2,6-二甲基吗啉基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(530mg，1.0mmol，83.3％yield)由中间体(E)-4-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)-2,6-二甲基吗啉(420mg，1.24mmol)和5-氯-2-甲基-4-[2-甲基-3-(4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)-苄氧基]-苯甲醛(480mg，1.20mmol)按照实施例17中的类似步骤制备而成。
MS Calcd:513.2MS Found:514.4([M+H]
+)
实施例69
(2S)-1-(5-氯-4-((3’-((E)-4-(2,6-二甲基吗啉基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
浅黄色固体(2S)-1-(5-氯-4-((3’-((E)-4-(2,6-二甲基吗啉基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(14.0mg,0.021mmol,10.8％yield)由中间体(E)-5-氯-4-((3’-(4-(2,6-二甲基吗啉)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(105mg，0.20mmol)和L-哌啶-2-甲酸(118.0mg,0.92mmol)按照实施例18似步骤制备而成。
1H-NMR(400MHz,DMSO-d
6)δppm 7.50(d,J＝7.6Hz,1H),7.42(d,J＝7.7Hz,1H),7.37(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.09(s,1H),7.06(d,J＝7.6Hz,1H),6.96(d,J＝7.4Hz,1H),6.72(d,J＝15.6Hz,1H),6.14(dt,J＝12.8,6.0Hz,1H)，5.18(s,2H),3.81(d,J＝13.5Hz,1H,),3.59-3.49(m,2H),2.82-2.72(m,2H),2.46-2.36(m,4H),2.30(s,3H),2.00(s,3H),1.94(s,3H),1.78(s,3H),1.69-1.57(m,4H),1.55-1.50(m,2H),1.42-1.37(m,2H),1.29-1.20(m,2H),1.05(s,3H),1.04(s,3H).
MS Calcd:644.3MS Found:645.5([M+H]
+).
实施例70
3-羟基哌啶-2-甲酸甲酯的制备
将3-羟基吡啶-2-甲酸(695.0mg,5.0mmol)溶解于甲醇(25mL)，冰浴下滴加SOCl
2(3.6mL，50.0mmol)，滴毕，加热至70℃反应18h，反应完毕，旋干溶剂，得到3-羟基吡啶-2-甲酸甲酯粗品810mg；取粗品120mg溶解于甲醇(15mL),加入Pt
2O(100mg)氢气条件下催化加氢反应24h，抽滤，滤液加Pd/C(5％,100mg)，氢气条件下继续催化加氢反应16h，抽滤，旋干滤液，得到油状物110mg，直接用于下一步反应。
MS Calcd:159.0MS Found:160.0([M+H]
+).
实施例71
(E)-4-(4-(3’-((2-氯-4-(氯甲基)-5-甲基苯氧基)甲基)-2,2’-二甲基-[1,1’-二苯基]-3-基)丁-3-烯-1-基)吗啉的制备
浅黄色固体(E)-5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(200mg，0.4mol)溶解于甲醇(20mL)，加入NaBH
4(75.6mg，2.0mmol)，室温反应2h，反应完毕，加水(50mL)稀释，二氯甲烷萃取(60mL),旋干溶剂，得到粗品中间体醇，溶解于SOCl
2(7mL),加热至70℃,反应1h，反应完毕，旋干SOCl
2,得到浅黄固体(E)-4-(4-(3’-((2-氯-4-(氯甲基)-5-甲基苯氧基)甲基)-2,2’-二甲基-[1,1’-二苯基]-3-基)丁-3-烯-1-基)吗啉，130mg，直接用于下一步反应。
MS Calcd:523.2MS Found:524.1([M+H]
+).
实施例72
(E)-1-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)3-羟基哌啶-2-羧酸
以上浅黄固体(130mg)溶解于乙腈(15mL)，加入碳酸钾(415mg，3.0mmol)，3-羟基哌啶-2-甲酸甲酯(粗品100mg)，反应加热至80℃，搅拌5小时，反应完毕，二氯甲烷：甲醇(5:1，60mL)萃取，水洗，干燥旋干，刮大板纯化，展开剂(二氯甲烷：甲醇＝5:1)，得到中间体45mg，溶解于四氢呋喃(6mL)加入氢氧化锂溶液(1.0mol/L，3.0mL)，反应16h，反应完毕，旋去四氢呋喃，加水稀释(5mL)，调节pH至弱酸性，析出白色固体，抽滤，烘干，得到(E)-1-(5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)3-羟基哌啶-2-羧酸(35mg，收率22.2％)。
1H NMR(400MHz,CDCl
3):δ7.51(d,J＝7.6Hz,1H),7.41(d,J＝7.6Hz,1H),7.37(s,1H),7.26(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.15(s,1H),7.01(d,J＝7.6Hz,1H),6.95(d,J＝7.6Hz,1H),6.70(d,J＝16.0Hz,1H),6.10-6.17(m,1H),5.20(s,2H),3.72(d,J＝13.6Hz,1H),3.50-3.62(m,8H),3.23-3.29(m,2H),2.99(s,1H),2.95(m,1H),2.40-2.47(m,6H),2.39(s,3H),2.05(s,3H),1.95(s,3H),1.65-1.75(m,2H),1.23-1.41(m,2H).
MS Calcd:632.3MS Found:633.4([M+H]
+).
实施例73
(E)-1-溴-2-甲基-3-(4-(甲基磺酰基)丁-1-烯-1-基)苯的制备
将(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(602mg，2.0mmol),甲烷亚磺酸钠(408.0mg,4.0mmol)和KI(342.0mg,2.0mmol)溶于N,N-二甲基甲酰胺(20mL)，加热至120℃，反应6h。加水(60mL)稀释，乙酸乙酯(60mL)萃取，水洗两遍，再饱和食盐水洗，有机相干燥旋干，用硅胶柱层析纯化(PE/EA＝30/1至10/1)，得到浅黄色油状物即中间体(E)-1-溴-2-甲基-3-(4-(甲基磺酰基)丁-1-烯-1-基)苯 (450mg,1.49mmol,74.7％yield)。
MS Calcd:302.0，304.0MS Found:302.9，305.0([M+H]
+).
实施例74
(E)-5-氯-4-((2,2’-二甲基-3’-(4-(甲基磺酰基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛的制备
(E)-5-氯-4-((2,2’-二甲基-3’-(4-(甲基磺酰基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(310mg，0.62mmol，41.9％yield)由中间体(E)-1-溴-2-甲基-3-(4-(甲基磺酰基)丁-1-烯-1-基)苯(450mg，1.49mmol)和5-氯-2-甲基-4-[2-甲基-3-(4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)-苄氧基]-苯甲醛(800mg，2.0mmol)按照实施例17中的类似步骤制备而成。
MS Calcd:496.1MS Found:497.2([M+H]
+)
实施例75
(S,E)-1-(5-氯-4-((2,2’-二甲基-3’-((E)-4-(甲基磺酰基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸的制备
浅黄色固体(S,E)-1-(5-氯-4-((2,2’-二甲基-3’-((E)-4-(甲基磺酰基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)哌啶-2-羧酸(17.0mg,0.028mmol,13.9％yield)由中间体(E)-5-氯-4-((2,2’-二甲基-3’-(4-(甲基磺酰基)丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(100mg，0.20mmol)和L-哌啶-2-甲酸(118.0mg,0.92mmol)按照实施例18似步骤制备而成。
1H-NMR(400MHz,DMSO-d
6)δppm 7.51(d,J＝7.5Hz,1H),7.45(d,J＝7.8Hz,1H),7.34(s,1H),7.29(t,J＝7.5Hz,1H),7.23(t,J＝7.6Hz,1H),7.13(s,1H), 7.07(d,J＝7.6Hz,1H),6.99(d,J＝7.3Hz,1H),6.81(d,J＝15.7Hz,1H),6.19(dt,J＝14.8,6.8Hz,1H)，5.20(s,2H),3.75(d,J＝13.4Hz,1H,),3.50-3.45(m,2H),3.01(s,3H),2.86-2.78(m,2H),2.73-2.61(m,2H),2.31(s,3H),2.19-2.09(m,2H),2.01(s,3H),1.97(s,3H),1.75-1.69(m,2H),1.46-1.41(m,4H).
MS Calcd:609.2MS Found:610.4([M+H]
+).
实施例76
(E)-N-(1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶-4-基)乙酰胺的制备
白色固体中间体(E)-N-(1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶-4-基)乙酰胺(220mg,0.60mmol,60％yield)是由(E)-1-溴-3-(4-溴丁-1-烯-1-基)-2-甲基苯(302mg,1.0mmol)和N-哌啶-4-基-乙酰胺(284mg,2.0mmol)按照实施例14中的类似步骤制备而成。
MS Calcd:364.1，366.1MS Found:365.0，367.2([M+H]
+).
实施例77
(E)-N-(1-(4-(3’-((2-氯-4-甲酰基-5-甲基苯氧基)甲基)-2,2’-二甲基-[1,1’-二苯基]-3-基)丁-3-烯-1-基)哌啶-4-基)乙酰胺的制备
中间体(E)-N-(1-(4-(3’-((2-氯-4-甲酰基-5-甲基苯氧基)甲基)-2,2’-二甲基-[1,1’-二苯基]-3-基)丁-3-烯-1-基)哌啶-4-基)乙酰胺(235mg，0.42mmol，70.1％yield)由中间体(E)-N-(1-(4-(3-溴-2-甲基苯基)丁-3-烯-1-基)哌啶-4-基)乙酰胺(450mg，1.49mmol)和5-氯-2-甲基-4-[2-甲基-3-(4,4,5,5-四甲基-[1,3,2]二氧硼杂环戊烷-2-基)-苄氧基]-苯甲醛(400mg，1.0mmol)按照实施例17中的类似步骤制备而成。
MS Calcd:558.2MS Found:559.3([M+H]
+)
实施例78
(S,E)-1-(4-((3’-(4-(4-乙酰胺基哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-5-氯-2-甲基苄基)哌啶-2-羧酸的制备
白色固体(S,E)-1-(4-((3’-(4-(4-乙酰胺基哌啶-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-5-氯-2-甲基苄基)哌啶-2-羧酸(15.0mg，0.022mol，12.4％yield)由中间体(E)-N-(1-(4-(3’-((2-氯-4-甲酰基-5-甲基苯氧基)甲基)-2,2’-二甲基-[1,1’-二苯基]-3-基)丁-3-烯-1-基)哌啶-4-基)乙酰胺(100mg，0.18mmol)和L-哌啶-2-甲酸(118.0mg,0.92mmol)按照实施例18似步骤制备而成。
1H-NMR(400MHz,DMSO-d
6)δppm 7.73(d,J＝7.7Hz,1H),7.51(d,J＝7.6Hz,1H),7.43(d,J＝7.8Hz,1H),7.35(s,1H),7.28(t,J＝7.6Hz,1H),7.21(t,J＝7.6Hz,1H),7.12(s,1H),7.07(d,J＝7.5Hz,1H),6.96(d,J＝7.4Hz,1H),6.71(d,J＝15.7Hz,1H),6.14(dt,J＝14.6,6.4Hz,1H),5.19(s,2H),3.77(d,J＝13.5Hz,1H),3.58(s,2H),3.26(d,J＝13.2Hz,1H),2.97-2.71(m,3H),2.42(d,J＝6.2Hz,1H),2.41-2.34(m,2H)2.30(s,3H),2.01(s,3H),1.95(s,3H),1.86(s,2H),1.77(s,3H),1.74-1.64(m,4H),1.53-1.48(m,1H),1.44-1.39(m,3H),1.38-1.28(m,4H)。
MS Calcd:671.3MS Found:672.4([M+H]
+).
实施例79
(E)-2-((5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)氨基)乙基-1-醇的制备
白色固体(E)-2-((5-氯-4-((2,2’-二甲基-3’-(4-吗啉丁-1-烯-1-基)-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苄基)氨基)乙基-1-醇(24.0mg，0.043mol，23.0％yield)由5-氯-4-((3’-(4-(吗啉-1-基)丁-1-烯-1-基)-2,2’-二甲基-[1,1’-二苯基]-3-基)甲氧基)-2-甲基苯甲醛(95mg，0.19mmol)和乙醇胺(122.0mg，2.0mmol)按照实施例6的类似步骤制备而成。
1H-NMR(400MHz,DMSO-d
6)δppm 7.67(s,1H),7.51(d,J＝7.8Hz,2H),7.34-7.20(m,3H),7.08(d,J＝7.7Hz,1H),7.00(d,J＝7.4Hz,1H),6.82(d,J＝15.7Hz,1H),6.16(dt,J＝15.5,6.8Hz,1H),5.27(s,1H),5.26(s,2H),4.10(s,2H),3.96(d,J＝12.4Hz,2H),3.86(t,J＝12.1Hz,2H),3.73(t,J＝5.4Hz,2H),3.45(d,J＝12.5Hz,2H),3.27-3.22(m,2H),3.13-3.01(m,5H),2.79-2.68(m,2H),2.41(s,3H),2.01(s,3H),1.97(s,3H).
MS Calcd:548.2MS Found:549.3[M+H]
+).
体外生物学评价
本检测方法用于本发明所述化合物的体外活性评价，包括体外蛋白水平结合抑制活性评价方法和细胞水平生物学功能活性评价方法。
本检测的目的在于综合评价不同化合物对PD-1/PD-L1结合抑制活性特点和对细胞模型的生物学活性影响，包括对特定模型细胞的生长活性以及对原代T细胞的生物活性影响。
实施例A 体外PD-1/PD-L1结合抑制筛选方法
实验主要原理(HTRF)
均相时间分辨荧光法(HTRF)：PD-1蛋白带His标签，其配体PD-L1的Fc融合蛋白带hFc标签。分别用Eu元素的鳌合标记物标记的抗hFc抗体和XL665标记的抗His抗体与两个对应标签结合，激光激发(320nm)后，能量能够从供体Eu上，转移到受体XL665，使得XL665发光(665nm)。当加入小分子化合物抑制剂后，阻断PD-1与PD-L1的结合，使得Eu和XL665距离较远，能量不能转移，XL665不会发光。
实验材料与设备
带His标签的人重组PD-1蛋白(Cat#：80448-R08H-100)、人重组PD-L1-Fc融合蛋白(Cat#：90251-C02H-100)购自义翘神州(Sino Biological Inc.)公司， 抗hFc-Eu
3+、抗His-XL665抗体购自Cisbio公司，其它相关试剂如稀释缓冲液、检测缓冲液等均从Cisbio公司购买。荧光检测仪器Tecan(Spark 10M)从瑞士Tecan公司购买。
实验主要过程
实验过程按照检测试剂使用说明书要求的流程进行(Invitrogen)。流程如下：
(1)实验准备：用稀释缓冲液将测试化合物稀释成不同浓度梯度(在20μL最终反应体系中最高终浓度为10μM)，将His-PD-1蛋白稀释成800nM(在20μL最终反应体系中终浓度100nM)，PD-L1-Fc融合蛋白稀释成16nM(终浓度为2nM)；用检测稀释液分别按试剂要求将抗His-XL665抗体和抗hFc-Eu
3+抗体稀释20倍和100倍。
(2)先将5μL测试化合物、2.5μLPD-L1蛋白和2.5μL PD-1蛋白溶液混匀后，室温反应15min；随后向该体系加入5μL anti-His-XL665抗体和5μL anti-hFc-Eu
3+抗体，继续孵育3h后检测。
(3)检测反应同时设置有对照组，包括未添加测试化合物的0抑制阳性对照、未添加PD-1蛋白的阴性对照。所有检测采用复孔进行。
(4)使用荧光检测仪Tecan(Spark 10M)检测每孔的荧光信号，激发波长为320nm，检测的发射波长分别为620nm和665nm。PD-1/PD-L1相互结合的强度参照荧光信号比值Em665/Em620。
(5)测试化合物的结合抑制率计算公式：抑制率(％)＝[1–(检测孔荧光信号比值–阴性对照)/(0抑制阳性对照荧光信号比值–阴性对照)×100％。不同浓度梯度的测试化合物分别计算结合抑制率后，进一步计算50％抑制浓度(IC
50)。数据见下表2。
表2 本发明代表性化合物抑制体外PD-1/PD-L1结合的IC
50数据
| 化合物 | IC 50(nM) | 化合物 | IC 50(nM) | 化合物 | IC 50(nM) |
| 5 | 120 | 10 | 14 | 13 | 29 |
| 16 | 15 | 19 | 17 | 22 | 20 |
| 23 | 4 | 25 | 36 | 27 | 2 |
| 29 | 3 | 30 | 56 | 33 | 27 |
| 36 | 12 | 41 | 11 | 47 | 38 |
| 50 | 5 | 53 | 15 | 62 | 8 |
| 63 | 1 | 65 | 4 | 66 | 8 |
| 69 | 26 | 72 | 16 | 75 | 16 |
| 78 | 3 | 79 | 3 |
由上述结果可见，本发明化合物具有良好的体外PD-1/PD-L1结合抑制活性。
Claims (27)
- 式I化合物，
或其可药用盐，其中，R 1选自氢、C 1-C 3烷基、CN、卤素、C 1-C 3卤代烷基；R 2选自氢、C 1-C 3烷基、CN、卤素、C 1-C 3烷氧基；L为任选取代的C 3-C 6烯基，所述C 3-C 6烯基中的一个CH 2任选地被-C(O)-、-S(O)-或-S(O) 2-所替代；B为氢，或NR 6R 7，或任选取代的具有1到3个独立地选自氮、氧或硫的杂原子的4元到7元饱和或部分不饱和杂环；其中R 6和R 7独立地为氢、C 1-C 3烷基、C 1-C 3卤代烷基、羟基C 1-C 3烷基或羧基C 1-C 3烷基；Z为氢、-CH 3、-CH＝CHAr或-OR 8；其中R 8选自氢、C 3-C 6烯基、卤代C 1-C 4烷基、羟基C 1-C 4烷基、-(CH 2) nX及-(CH 2) nAr；n为1、2、3或4；X选自氢、-CH 3、-CF 3、C 1-C 4烷氧基、-N(CH 3) 2、任选经一个或两个卤素取代的C 3-C 6环烷基、-CN、-CO 2R g、-C(O)NH 2、-C(O)N(CH 3) 2、吗啉基、四氢吡喃基、任选经羟基取代的吡咯烷酮基及任选经一个或两个独立选自以下的基团取代的哌啶基：C 1-C 4烷基、羧基、羟基及C 1-C 4烷氧基羰基；
R g选自氢及C 1-C 4烷基；及Ar选自苯并二噁烷基、吲唑基、异喹啉基、异噁唑基、萘基、噁二唑基、苯基、吡啶基、嘧啶基及喹啉基；其中各环任选经1、2、3或4个独立选自以下的取代基取代：C 1-C 4烷氧基、C 1-C 4烷氧基羰基、C 1-C 4烷氧基羰基氨基、C 1-C 4烷基、C 1-C 4烷基羰基、C 1-C 4烷基磺酰基、甲酰氨基、甲酰氨基C 1-C 4烷基、-(CH 2) qCO 2-(C 1-C 4烷基)、-(CH 2) qOH、羧基、氰基、甲 酰基、卤素、卤代C 1-C 4烷基、卤代C 1-C 4烷氧基、硝基、任选经一个氰基取代的苯基、任选经一个卤素取代的苯基氧基、苯基羰基、吡咯、四氢吡喃及
其中q为0、1、2、3或4且其中t、z及R z如下定义：t为0、1、2、3或4；z为1、2或3；各R z独立选自C 1-C 4烷氧基、C 1-C 4烷氧基羰基、C 1-C 4烷氧基羰基C 1-C 4烷基、C 1-C 4烷基、C 1-C 4烷基酰氨基、C 1-C 4烷基氨基、C 1-C 4烷基羰基、酰氨基、羧基、羧基C 1-C 4烷基、二(C 1-C 4烷基)酰氨基、二(C 1-C 4烷基)氨基、卤素、卤代C 1-C 4烷氧基、卤代C 1-C 4烷基、羟基、羟基C 1-C 4烷基、吗啉基、-NR cR d、(NR cR d)C 1-C 4烷基、-NR eR f、(NR eR f)C 1-C 4烷基、氧代、苯基及苯基C 1-C 4烷基，其中该苯基及该苯基C 1-C 4烷基的苯基部分任选经一个、两个或三个独立选自C 1-C 3烷基及卤素的基团取代；R c及R d独立选自氢、C 2-C 4烯基羰基、C 1-C 4烷氧基羰基、C 1-C 6烷基、C 1-C 4烷基羰基、酰氨基C 1-C 4烷基、氨基C 1-C 4烷基、芳基C 1-C 4烷基、C 3-C 10环烷基、(C 3-C 10环烷基)C 1-C 4烷基、卤代C 1-C 4烷基羰基、杂环基C 1-C 4烷基、杂环基C 1-C 4烷基羰基、羟基C 1-C 6烷基及羟基C 1-C 4烷基羰基，其中该酰氨基C 1-C 4烷基、该氨基C 1-C 4烷基、该芳基C 1-C 4烷基、该(C 3-C 10环烷基)C 1-C 4烷基、该杂环基C 1-C 4烷基及该杂环基C 1-C 4烷基羰基的烷基部分任选经一个或两个独立选自羧基及羟基的基团取代；其中该羟基C 1-C 4烷基及该羟基C 1-C 4烷基羰基的烷基部分任选经一个或两个独立选自羧基及羟基的基团取代；且其中该芳基C 1-C 4烷基的芳基部分、该C 3-C 10环烷基、该(C 3-C 10环烷基)C 1-C 4烷基的环烷基部分及该杂环基C 1-C 4烷基及该杂环基C 1-C 4烷基羰基的杂环基部分各任选经一个、两个或三个独立选自C 1-C 4烷氧基羰基、C 1-C 4烷基及卤素的基团取代；R e及R f与其所连接的原子一起形成选自吗啉及的环；R 3独立选自C 2-C 4烯基、C 1-C 4烷氧基、C 1-C 4烷基、氰基、卤素及卤代C 1-C 4 烷基；
m为0、1或2；R 4选自氢、C 1-C 4烷基、苄基、(C 3-C 6环烷基)C 1-C 3烷基、卤代C 1-C 4烷基、任选经第二个羟基取代的羟基C1-C6烷基及任选经氰基取代的吡啶基(C 1-C 3烷基)；及R 5选自氢、C 1-C 4烷基、-(CH 2) nN(CH 3) 2、羧基C 2-C 6烯基、羧基C 1-C 6烷基及羟基C 1-C 6烷基，其中该羧基C 1-C 6烷基及该羟基C 1-C 6烷基的烷基部分任选经一个羟基或苯基取代，其中该苯基进一步任选经羟基取代；或者，R 5选自以下基团：
其中R w为-CONH 2；R 10选自氢、苄基及甲基；各R 10’独立选自氢及C 1-C 3烷基；R 11选自氢、C 1-C 3烷基及苄基；R 12选自C 2-C 4烯基及C 1-C 4烷基；及R 50选自氢、C 1-C 6烷基及C 1-C 6烷氧基羰基；或R 4及R 5与其所连接的氮原子一起形成选自以下的环：其中
s为0、1或2；z为1、2或3；p为0，1或2；R 13及R 13’独立选自氢、羧基、羟基C 1-C 4烷基、氧代及-C(O)NHSO 2R 16；R 14独立选自氢、羟基C 1-C 4烷基、氧代及羧基；各R 15独立选自C 1-C 4烷氧基羰基、C 1-C 6烷基、羧基、卤素、羟基、羟基 C 1-C 4烷基、-NR c’R d’及苯基氧基羰基，其中该苯基任选经硝基取代，其中R c’及R d’独立选自氢、C 1-C 4烷氧基羰基及C 1-C 4烷基羰基；及R 16选自三氟甲基、环丙基、C 1-C 4烷基、二甲基氨基及经甲基取代的咪唑基。 - 权利要求1所述的式I化合物或其可药用盐，其中，R 1和R 2独立地选自氢和C 1-C 3烷基；L为-CH 2-CH＝CH-或-CH 2-CH 2-CH＝CH-，其中一个CH 2任选地被-C(O)-、-S(O)-或-S(O) 2-所替代，上述L定义中基团左侧与式I结构中B基团相连；Z为甲基、-CH＝CHAr或-O(CH 2) nAr；其中n为1、2、3或4；Ar选自苯并二噁烷基、吲唑基、异喹啉基、异噁唑基、萘基、噁二唑基、苯基、吡啶基、嘧啶基及喹啉基；其中各环任选经1、2、3或4个独立选自以下的取代基取代：C 1-C 4烷氧基、C 1-C 4烷氧基羰基、C 1-C 4烷氧基羰基氨基、C 1-C 4烷基、C 1-C 4烷基羰基、C 1-C 4烷基磺酰基、酰氨基、酰氨基C 1-C 4烷基、-(CH 2) qCO 2C 1-C 4烷基、-(CH 2) qOH、羧基、氰基、甲酰基、卤素、卤代C 1-C 4烷基、卤代C 1-C 4烷氧基、硝基、任选经一个氰基取代的苯基、任选经一个卤素取代的苯基氧基、苯基羰基、吡咯、四氢吡喃，其中q为0、1、2、3或4；R 3独立选自C 1-C 4烷氧基和卤素。
- 权利要求2所述的式I化合物或其药用盐，其中R 1和R 2独立选自甲基及卤素；Z为甲基、-CH＝CHAr或-OCH 2Ar；其中Ar为任选经一个或两个独立选自以下的基团取代的吡啶基：C 1-C 4烷基、C 1-C 4烷基磺酰基、甲酰氨基、氰基及卤素；R 3为卤素；且m为1。
- 权利要求1所述的式I化合物，其选自
或其可药用盐。 - 制备如权利要求1至4中任一项所述的式I化合物的方法，
包括使式A化合物
与式F化合物
在有机溶剂和还原剂的作用下经还原胺化形成式I化合物，其中R 1、R 2、R 3、R 4、R 5、L、B、Z和m如权利要求1中所定义。 - 根据权利要求5所述的制备方法，其中所述还原剂选自NaBH 4、NaBH 3CN及NaBH 3OAc。
- 根据权利要求5或6所述的制备方法，其中所述还原胺化反应在碱性催化剂存在下进行，并且所述碱性催化剂选自TEA和DIPEA。
- 根据权利要求5至7中任一项所述的制备方法，其中所述还原胺化反应在酸性催化剂存在下进行，并且所述酸性催化剂选自醋酸、盐酸和三氟醋酸。
- 根据权利要求5至8中任一项所述的制备方法，其中所述制备方法在干燥剂存在下进行，并且所述干燥剂选自无水MgSO 4、无水Na 2SO 4和分子筛。
- 根据权利要求5至9中任一项所述的制备方法，其中所述还原胺化反应在溶剂中进行，并且所述溶剂选自CH 3OH、CH 2Cl 2和1,2-二氯乙烷。
- 药物组合物，其包含有效量的权利要求1至4中任一项所述的式I化合物或其可药用盐，以及任选的可药用赋形剂或载体。
- 权利要求1至4中任一项所述的化合物或其可药用盐在制备用于提高、刺 激、调节和/或增加免疫应答的药物中的用途。
- 权利要求1至4中任一项所述的化合物或其可药用盐在制备用于抑制癌细胞生长、增殖或转移的药物中的用途。
- 权利要求13的用途，其中所述癌症选自鳞状非小细胞肺癌(NSCLC)、非鳞状NSCLC。
- 权利要求1至4中任一项所述的化合物或其可药用盐在制备用于治疗或预防感染性疾病的药物中的用途。
- 权利要求15的用途，其中所述感染性疾病是由病毒引起的感染性疾病。
- 权利要求1至4中任一项所述的化合物或其可药用盐在制备用于治疗或预防自身免疫性疾病的药物中的用途。
- 权利要求1至4中任一项所述的化合物或其可药用盐在制备用于治疗或预防败血性休克的药物中的用途。
- 式A化合物
其中R 1、R 2、R 3、L、B、Z和m如权利要求1中所定义。 - 制备如权利要求19所述的式A化合物的方法，包括使式5化合物
在将羟基转变为好的离去基团，如-OMs、-OTs或溴以后与式7化合物
反应得到式A化合物。 - 根据权利要求20所述的制备方法，其中羟基已转变为好的离去基团之后的式5化合物与式7化合物的反应在去酸剂存在下进行，并且所述去酸剂选自NaOH、K 2CO 3和KOH。
- 根据权利要求20至21中任一项所述的制备方法，其中羟基已转变为好的离去基团之后的式5化合物与式7化合物的反应在碱性条件下发生酚的O-烃化反应，并且所述酚的O-烃化反应在选自水、甲醇、乙醇、异丙醇、丙酮、DMF、DMSO、苯和二甲苯的溶剂中进行。
- 制备如权利要求19所述的式A化合物的方法，包括使式3化合物
与式8化合物
在金属催化作用下进行铃木交叉偶联反应。 - 根据权利要求23所述的制备方法，其中所述金属催化剂选自钯催化剂如Pd(PPh 3) 4、Pd(dffp)Cl 2、Pd(OAc) 2、Pd(dba) 3/PCy 3、PdCl 2、Pd(PPh 3) 4Cl 2，或者Ni催化剂如NiCl 2(dffp)、NiCl 2(dffp)/Zn、NiCl 2(dffp)/BuLi、NiCl 2(PPh 3) 2/PPh 3、NiCl 2(NEt 3) 2、NiCl 2(NEt 3) 2、NiCl(bipy)、Ni(TPPS) 3、Ni(COD) 2、NiCl 2(PPh 3) 2/n-BuLi、Ni{P(OMe) 3} 2Cl 2、NiCl 2(PCy 3) 2。
- 根据权利要求23至24中任一项的制备方法，其中所述铃木交叉偶联反应在碱存在下进行，并且所述碱为KOAc、K 3PO 4、K 2CO 3、NaOH、Ba(OH) 2、Na 2CO 3、CsF或NaHCO 3。
- 根据权利要求23至25中任一项的制备方法，其中所述铃木交叉偶联反应在溶剂中进行，并且所述溶剂选自乙醇、THF、异丙醇、DMSO、DMF、二氧六环、甲苯、水和DME。
- 权利要求19所述的式A化合物用作制备权利要求1所述式I化合物的中间体的用途。
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018009505A1 (en) * | 2016-07-08 | 2018-01-11 | Bristol-Myers Squibb Company | 1,3-dihydroxy-phenyl derivatives useful as immunomodulators |
| WO2018045142A1 (en) * | 2016-09-02 | 2018-03-08 | Polaris Pharmaceuticals, Inc. | Immune checkpoint inhibitors, compositions and methods thereof |
| WO2018044963A1 (en) * | 2016-09-01 | 2018-03-08 | Bristol-Myers Squibb Company | Biaryl compounds useful as immunomodulators |
| CN108368090A (zh) * | 2015-10-15 | 2018-08-03 | 百时美施贵宝公司 | 作为免疫调节剂的化合物 |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BR112016004194A8 (pt) * | 2013-09-04 | 2020-02-11 | Bristol Myers Squibb Co | compostos úteis como imunomoduladores |
| CN109311792B (zh) * | 2016-05-23 | 2022-10-21 | 中国医学科学院药物研究所 | 苯醚类衍生物、及其制法和药物组合物与用途 |
| WO2018121560A1 (zh) * | 2016-12-29 | 2018-07-05 | 深圳微芯生物科技有限责任公司 | 脲类化合物、其制备方法及其应用 |
| CN108268090A (zh) * | 2016-12-30 | 2018-07-10 | 维沃移动通信有限公司 | 一种接口保护装置、移动终端和方法 |
| US11787766B2 (en) * | 2017-08-18 | 2023-10-17 | Shanghai Ennovabio Pharmaceuticals Co., Ltd. | Compound having PD-L1 inhibitory activity, preparation method therefor and use thereof |
| CN109305934A (zh) * | 2018-08-07 | 2019-02-05 | 成都海博锐药业有限公司 | 苯醚类衍生物及可药用盐、医药上的用途 |
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2019
- 2019-07-30 CN CN201910695833.7A patent/CN110872275A/zh active Pending
- 2019-08-28 TW TW108130887A patent/TWI712600B/zh active
- 2019-08-29 WO PCT/CN2019/103225 patent/WO2020043154A1/zh active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108368090A (zh) * | 2015-10-15 | 2018-08-03 | 百时美施贵宝公司 | 作为免疫调节剂的化合物 |
| WO2018009505A1 (en) * | 2016-07-08 | 2018-01-11 | Bristol-Myers Squibb Company | 1,3-dihydroxy-phenyl derivatives useful as immunomodulators |
| WO2018044963A1 (en) * | 2016-09-01 | 2018-03-08 | Bristol-Myers Squibb Company | Biaryl compounds useful as immunomodulators |
| WO2018045142A1 (en) * | 2016-09-02 | 2018-03-08 | Polaris Pharmaceuticals, Inc. | Immune checkpoint inhibitors, compositions and methods thereof |
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| TWI712600B (zh) | 2020-12-11 |
| TW202033514A (zh) | 2020-09-16 |
| CN110872275A (zh) | 2020-03-10 |
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