WO2019076343A1 - Pd-1/pd-l1小分子抑制剂及其制备方法和用途 - Google Patents

Pd-1/pd-l1小分子抑制剂及其制备方法和用途 Download PDF

Info

Publication number
WO2019076343A1
WO2019076343A1 PCT/CN2018/110869 CN2018110869W WO2019076343A1 WO 2019076343 A1 WO2019076343 A1 WO 2019076343A1 CN 2018110869 W CN2018110869 W CN 2018110869W WO 2019076343 A1 WO2019076343 A1 WO 2019076343A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
group
formula
ring
substituted
Prior art date
Application number
PCT/CN2018/110869
Other languages
English (en)
French (fr)
Inventor
王喆
范国钦
曾志宏
王晓光
江荣珍
Original Assignee
上海长森药业有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 上海长森药业有限公司 filed Critical 上海长森药业有限公司
Publication of WO2019076343A1 publication Critical patent/WO2019076343A1/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/12—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A61K31/343—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—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 ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4365—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 ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system having sulfur as a ring hetero atom, e.g. ticlopidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/498—Pyrazines or piperazines ortho- and peri-condensed with carbocyclic ring systems, e.g. quinoxaline, phenazine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4985—Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/58—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems with hetero atoms directly attached to the ring nitrogen atom
    • C07D215/60—N-oxides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
    • C07D239/72—Quinazolines; Hydrogenated quinazolines
    • C07D239/74—Quinazolines; Hydrogenated quinazolines with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached to ring carbon atoms of the hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/36—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems
    • C07D241/38—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems with only hydrogen or carbon atoms directly attached to the ring nitrogen atoms
    • C07D241/40—Benzopyrazines
    • C07D241/42—Benzopyrazines with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
    • C07D307/79—Benzo [b] furans; Hydrogenated benzo [b] furans with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/76—Benzo[c]pyrans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D319/00—Heterocyclic compounds containing six-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D319/10—1,4-Dioxanes; Hydrogenated 1,4-dioxanes
    • C07D319/14—1,4-Dioxanes; Hydrogenated 1,4-dioxanes condensed with carbocyclic rings or ring systems
    • C07D319/16—1,4-Dioxanes; Hydrogenated 1,4-dioxanes condensed with carbocyclic rings or ring systems condensed with one six-membered ring
    • C07D319/20—1,4-Dioxanes; Hydrogenated 1,4-dioxanes condensed with carbocyclic rings or ring systems condensed with one six-membered ring with substituents attached to the hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
    • C07D333/52—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes
    • C07D333/54—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • the present invention belongs to the field of pharmaceutical synthesis, and in particular, the present invention relates to an inhibitor of PD-1/PD-L1, a preparation method thereof and use thereof.
  • Programmed death-1 PD-1 is a 50-55KD type I transmembrane glycoprotein belonging to the immunoglobulin superfamily. Its remarkable feature is that the cytoplasmic region contains N-terminus and C-, respectively. Two tyrosine residues at the end. The former is involved in the formation of an immunoreceptor tyrosine inhibition motif (ITIM), which is involved in the formation of an immunoreceptor tyrosine transfer motif (ITSM). After PD-1 binds to the ligand, the tyrosine in the ITSM region is phosphorylated, and the protein tyrosine phosphatase molecule is recruited, which dephosphorylates the downstream effector molecule and transduces the negative signal to play a negative regulatory inhibition. Proliferation of T cells and production of cytokines.
  • ITIM immunoreceptor tyrosine inhibition motif
  • ITSM immunoreceptor tyrosine transfer motif
  • PD-1 is present on the cell surface in monomeric form, and is the first to express double negative cells in the thymus. It can also be expressed in activated T cells, B cells, natural killer cells, dendritic cells, and activated monocytes. PD-1 usually forms a signaling pathway together with its ligand, and its ligands are PD-L1 and PD-L2, which have 40% identical amino acid sequences. The extracellular region has IgC and IgV-type domains, which are similar in structure but different in distribution. The PD-L1 distribution is widely expressed in murine T cells, B cells, dendritic cells, macrophages and stem cell cultured bone marrow cells.
  • PD-1 Although the nucleotide sequence of human and murine PD-1 has 70% homology, both encode a protein consisting of 288 amino acid residues and 60% homology at the amino acid level, but human PD-L1 The expression is lower than the mouse source.
  • the relative limitation of PD-L2 distribution is mainly expressed in activated monocyte macrophages and dendritic cells.
  • PD-1 and its ligand (mainly PD-L1) have a negative immunomodulatory effect.
  • the PD-1/PD-L1 signaling pathway can induce and maintain immune tolerance in peripheral tissues, and has a positive effect on preventing excessive inflammatory reactions of tissues and the occurrence of autoimmune diseases.
  • this signaling pathway also plays an important regulatory role in antiviral or antitumor.
  • PD-L1 is expressed in tumor cells as well as in immune cells, and binding of PD-1 to it reduces the production of T cells in cancer patients, inhibits anti-tumor immune responses and leads to depletion of T cells.
  • viruses use similar mechanisms to disrupt effective immune recognition.
  • the current PD-1/PD-L1 series of drugs are primarily monoclonal antibodies and have shown impressive clinical conditions, including melanoma and lung cancer.
  • monoclonal antibodies to both PD-1 inhibitors are marketed under FDA approval, including merck's pembrolizumab and BMS nivolumab.
  • Roche's PD-L1 inhibitor Atezolizumab was also approved by the FDA for use as a second-line drug for the treatment of advanced bladder cancer.
  • monoclonal antibodies also exhibit significant disadvantages such as very limited tissue and tumor penetration, long half-life, lack of oral bioavailability, and difficult and expensive production.
  • current targeting of PD-1/PD-L1 monoclonal antibodies is only responsive in a small number of cases and tumor types.
  • the compounds described herein are capable of inhibiting the biological activity of PD-1/PD-L1 and have therapeutic and research uses.
  • ring A is formed by ring B and ring C, as shown by formula L-1 and formula L-2:
  • Ring B and Ring C are each independently a 5-6 membered aromatic ring, a 5-6 membered aromatic heterocyclic ring, a 5-6 membered saturated aliphatic ring or a saturated or 5-6 membered aliphatic heterocyclic ring; and said ring B and ring C constitutes a parallel structure;
  • X is absent (ie, R 5 is directly attached to ring C or ring B), 5-10 membered aryl or heteroaryl;
  • R 5 is an unsubstituted or substituted group selected from the group consisting of a 5-10 membered aryl group, a 5-10 membered heteroaryl group, a 5-10 membered ring group, and a 5-10 membered heterocyclic group; Substituted by one or more substituents selected from the group consisting of C1-C6 alkyl, halo C1-C6 alkyl, carboxy (-COOH), ester (C1-C6 alkyl O(CO) -);
  • heterocyclic group, heteroaryl group, aromatic heterocyclic ring or saturated aliphatic heterocyclic ring contains 1-3 (1, 2 or 3) hetero atoms selected from N, S and O;
  • substitution means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6.
  • R 5 -X- is a structure selected from the group consisting of:
  • R 10 , R 11 , R 12 , R m , R n , R L are each independently selected from the group consisting of H, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkane Oxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo, -CN, hydroxy, amino, carboxy, C6-C10 aryl, halogenated a C6-C10 aryl group, a 5-10 membered heteroaryl group having 1-3 hetero atoms selected from N, S and O, a halogenated one having 1-3 hetero atoms selected from N, S and O 5-10 yuan heteroaryl.
  • the compound of formula L is selected from the group consisting of
  • the compound of formula L is selected from the group consisting of
  • each group has the same definition as before.
  • the compound is selected from the group consisting of:
  • the compound is a compound of formula I-5, the method comprising the steps of:
  • the compound is a compound of formula V-7, the method comprising the steps of:
  • the compound is a compound of the formula VI-8, the method comprising the steps of:
  • the compound is a compound of the formula VII-4, the method comprising the steps of:
  • the compound is a compound of formula VIII-8, the method comprising the steps of:
  • the compound is a compound of formula X-6, the method comprising the steps of:
  • the compound of (11) is a compound of formula XI-7, and the method comprises the steps of:
  • the compound of (12) is a compound of the formula XII-7, the method comprising the steps of:
  • each group has the same definition as before.
  • an intermediate represented by the following formula, a stereoisomer, a cis-trans isomer or a tautomer thereof,
  • R 1 , R 2 , R 3 , R 4 and R 5 have the same meanings as defined above.
  • a pharmaceutical composition comprising (1) a compound according to the first aspect of the invention, a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof , hydrate or solvate; (2) a pharmaceutically acceptable carrier.
  • a compound, a stereoisomer thereof or a tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutically acceptable salt thereof Use of the pharmaceutical composition of the third aspect of the invention, characterized in that it is used for the preparation of a PD-1/PD-L1 inhibitor.
  • a compound according to the first aspect of the invention a stereoisomer thereof or a tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or The use of the pharmaceutical composition of the third invention thereof, characterized in that it is used for the preparation of a medicament for treating cancer; preferably, the cancer is selected from the group consisting of melanoma, lung cancer, bladder cancer.
  • a compound according to the first aspect of the invention a stereoisomer thereof or a tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or The use of the pharmaceutical composition of the third invention thereof, characterized in that it is used for the preparation of a pharmaceutical composition for treating or preventing a disease associated with PD-1/PD-L1 activity or expression.
  • Another aspect of the invention provides a method of treatment comprising the steps of administering a compound according to the first aspect of the invention, a stereoisomer thereof or a tautomer thereof, or a pharmaceutical thereof, to a patient in need thereof
  • An acceptable salt, hydrate or solvate or a pharmaceutical composition according to the third aspect of the invention is an acceptable salt, hydrate or solvate or a pharmaceutical composition according to the third aspect of the invention.
  • the term “about” means that the value can vary by no more than 1% from the recited value.
  • the expression “about 100” includes all values between 99 and 101 and (eg, 99.1, 99.2, 99.3, 99.4, etc.).
  • the terms "containing” or “including” may be open, semi-closed, and closed. In other words, the terms also include “consisting essentially of,” or “consisting of.”
  • alkyl as used herein includes a straight or branched alkyl group.
  • C 1 -C 8 alkyl represents a straight or branched alkyl group having 1-8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl. Wait.
  • alkenyl as used herein, includes a straight or branched alkenyl group.
  • C 2 -C 6 alkenyl refers to a straight or branched alkenyl group having 2 to 6 carbon atoms, such as ethenyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2 a butenyl group, or a similar group.
  • alkynyl includes a straight or branched alkynyl group.
  • C 2 -C 6 alkynyl refers to a straight or branched alkynyl group having 2 to 6 carbon atoms, such as an ethynyl group, a propynyl group, a butynyl group, or the like.
  • C 3 -C 8 cycloalkyl means a cycloalkyl group having 3-8 carbon atoms. It may be a monocyclic ring such as a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, or the like. It may also be in the form of a double loop, such as a bridged or spiro ring.
  • C 1 -C 8 alkoxy refers to a straight or branched alkoxy group having from 1 to 8 carbon atoms; for example, methoxy, ethoxy, propoxy, iso Propyloxy, butoxy, isobutoxy, tert-butoxy and the like.
  • the term "3-10 membered heterocycloalkyl having 1-3 heteroatoms selected from the group consisting of N, S and O" means having from 3 to 10 atoms and wherein 1-3 of the atoms are A saturated or partially saturated cyclic group selected from the group consisting of heteroatoms of N, S and O. It may be a single ring or a double ring form, such as a bridge ring or a spiro ring. Specific examples may be oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl, pyrrolidinyl and the like.
  • C 6 -C 10 aryl refers to an aryl group having 6 to 10 carbon atoms, for example, a phenyl or naphthyl group or the like.
  • the term "5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S and O" refers to having 5-10 atoms and wherein 1-3 atoms are selected from A cyclic aromatic group of the following group of heteroatoms of N, S and O. It may be a single ring or a fused ring.
  • Specific examples may be pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2, 4)-Triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl and the like.
  • the groups of the present invention may be substituted with a substituent selected from the group consisting of halogen, nitrile, nitro, hydroxy, amino, C 1 -C 6 alkyl-amino, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkyl, halogenated C 2 -C 6 alkenyl, halogen C 2 -C 6 alkynyl, halo C 1 -C 6 alkoxy, allyl, benzyl, C 6 -C 12 aryl, C 1 -C 6 alkoxy-C 1 -C 6 alkane , C 1 -C 6 alkoxy-carbonyl, phenoxycarbonyl, C 2 -C 6 alkynyl-carbonyl, C 2 -C 6 alkenyl-carbonyl, C 3 -C
  • halogen or halogen atom refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from the group consisting of F, Cl and Br. "Halo” means substituted with an atom selected from the group consisting of F, Cl, Br, and I.
  • the structural formulae described herein are intended to include all isomeric forms (such as enantiomeric, diastereomeric, and geometric isomers (or conformational isomers): for example, containing asymmetry The central R, S configuration, the (Z) and (E) isomers of the double bond.
  • a single stereochemical isomer of a compound of the invention, or a mixture of enantiomers, diastereomers or geometric isomers (or conformational isomers) thereof, is within the scope of the invention.
  • tautomer means that structural isomers having different energies can exceed the low energy barrier and thereby transform each other.
  • proton tautomers ie, proton shifts
  • proton transfer such as 1H-carbazole and 2H-carbazole.
  • Valence tautomers include interconversion through some bonding electron recombination.
  • solvate refers to a complex of a compound of the invention that is coordinated to a solvent molecule to form a particular ratio.
  • hydrate refers to a complex formed by the coordination of a compound of the invention with water.
  • a compound of the invention refers to a compound of formula (L), and also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula (L).
  • pharmaceutically acceptable salt refers to a salt of the compound of the invention formed with an acid or base suitable for use as a medicament.
  • Pharmaceutically acceptable salts include inorganic and organic salts.
  • a preferred class of salts are the salts of the compounds of the invention with acids.
  • Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, Organic acids such as maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzoic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.
  • mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid,
  • Organic acids such as maleic acid, lactic acid, malic acid, tartaric acid,
  • the compound of the present invention has excellent inhibitory activity against PD-1, the compound of the present invention and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the present invention are mainly active.
  • the pharmaceutical composition of the composition can be used to prevent and/or treat diseases associated with the PD-1/PD-L1 signaling pathway (eg, cancer).
  • compositions of the present invention comprise a safe and effective amount of a compound of the invention and a pharmaceutically acceptable excipient or carrier.
  • safe and effective amount it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects.
  • the pharmaceutical compositions contain from 1 to 2000 mg of the compound of the invention per agent, more preferably from 10 to 200 mg of the compound of the invention per agent.
  • the "one dose” is a capsule or tablet.
  • “Pharmaceutically acceptable carrier” means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By “compatibility” it is meant herein that the components of the composition are capable of intermixing with the compounds of the invention and between them without significantly reducing the potency of the compound.
  • pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid).
  • magnesium stearate magnesium stearate
  • calcium sulfate vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as Tween ), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.
  • vegetable oil such as soybean oil, sesame oil, peanut oil, olive oil, etc.
  • polyol such as propylene glycol, glycerin, mannitol, sorbitol, etc.
  • emulsifier such as Tween
  • a wetting agent such as sodium lauryl sulfate
  • a coloring agent such as a flavoring agent, a stabilizer, an antioxidant, a preservative
  • the mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, parenteral (intravenous, intramuscular or subcutaneous).
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and
  • Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs.
  • the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.
  • inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethyl
  • compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
  • the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • suspending agents for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.
  • compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion.
  • Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.
  • the compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds.
  • the pharmaceutical composition When administered in combination, the pharmaceutical composition also includes one or more other pharmaceutically acceptable compounds.
  • One or more of the other pharmaceutically acceptable compounds can be administered simultaneously, separately or sequentially with the compounds of the invention.
  • a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight
  • the dose to be administered is usually from 1 to 2000 mg, preferably from 20 to 500 mg.
  • specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.
  • the compounds of the present invention are novel in structure and have excellent PD-1/PD-L1 inhibition.
  • the nuclear magnetic resonance spectrum was analyzed by a Bruker AV-400 (400 MHz) nuclear magnetic instrument. Chemical shifts tetramethylsilane as an internal standard to record, expressed in ppm (CDC1 3: ⁇ 7.26ppm). The recorded data information is as follows: chemical shift and its split and coupling constant (s: singlet; d: doublet; t: triplet; q: quartet; br: broad; m: multiplet). Mass spectrometry data was analyzed using a liquid-mass spectrometer from Finnigan LCQ Advantage.
  • the raw materials and reagents according to the present invention can be purchased commercially or custom-made, unless otherwise specified.
  • the compounds of the invention may contain one or more asymmetric centers, such that the series of compounds may be in racemic or single enantiomeric form.
  • the compound prepared by the present invention (Formula IV) is a heterocyclic compound having a purity higher than 95%, and the structural characterization of each final product is determined by MS or/and hydrogen spectrum nuclear magnetic resonance ( 1 H NMR) analysis, respectively.
  • MS or/and hydrogen spectrum nuclear magnetic resonance ( 1 H NMR) analysis MS or/and hydrogen spectrum nuclear magnetic resonance ( 1 H NMR) analysis, respectively.
  • Step 1-2
  • Step 2-1
  • Step 2-2
  • the substrate (1 g) was dissolved in 30 mL of chloroform and heated to 45 ° C, and liquid bromine (0.83 g) was slowly added dropwise. After the addition was completed, TLC showed the reaction was complete. After cooling, the sodium thiosulfate solution was washed, and the organic phase was washed with water and a saturated sodium chloride solution and dried over anhydrous sodium sulfate. After concentration, column chromatography gave 0.9 g of pale yellow solid.
  • Step 3-2
  • Triphenylphosphine (3.0 eq.) was added to a solution of CBr 4 (1.5 eq.) in dichloromethane at 0 ° C, and the reaction was stirred for 15 min.
  • the methylene chloride was concentrated, filtered, concentrated and purified by column chromatography to give a dibromo compound.
  • n-Butyllithium (2.5 M, hexane) was added dropwise to a solution of the dibromo compound in THF at 0 ° C. After the addition was completed, the temperature was maintained and the mixture was stirred for 0.5 h. The temperature was then raised to room temperature for 1.5 h, and TLC detection showed the reaction was complete. The reaction was quenched by dropwise addition of water.
  • Step 5-2
  • the substrate ketone (0.68 g) was dissolved in THF, NaH (26 mg) and dichloropiperidine (0.48 g) were added, and the mixture was heated to reflux for 5 h under argon atmosphere, and refluxed without addition of 0.2 g of NaH. After cooling, the reaction was quenched by water, extracted with ethyl acetate and dried over anhydrous sodium sulfate. After concentrating, the crude product was dissolved in DMF, and then added with potassium carbonate (0.85 g). The mixture was heated to 100 ° C for 16 h. After cooling, DMF was concentrated under reduced pressure, and water and ethyl acetate were evaporated. Chromatography gave 440 mg of solid.
  • Step 6-2
  • the white solid 54 obtained in the previous reaction was dissolved in dichloromethane, and phosphorus oxychloride and a catalytic amount of DMF were added under argon atmosphere. After the reaction overnight, TLC showed that the substrate was completely reacted. After quenching the reaction with water, the organic phase was dried over anhydrous sodium sulfate. After concentration, column chromatography gave 60 mg of solid.
  • Step 6-3
  • Step 6-4
  • the intermediate (5 g) was dissolved in 50 mL of chlorobenzene, 5 g of polyphosphoric acid was added, and the mixture was heated to 110 ° C overnight, and TLC showed the reaction was completed. After the temperature was lowered, a 10% sodium hydroxide solution was added dropwise to adjust the pH to 7, and water and ethyl acetate were added, and the organic phase was washed with water and brine, and dried over anhydrous sodium sulfate. Column chromatography gave 2.43 g of a white solid.
  • the crude product was dissolved in methanol, and sodium borohydride (temperature less than 20 deg.) was added portionwise in an ice water bath. After the addition, the mixture was stirred for 1/2 h. Excess sodium borohydride was quenched by the addition of acetone. Then tetrahydrofuran and 6N hydrochloric acid solution were added until pH ⁇ 1. The reaction solution was heated to 40 ° C for 0.5 h, and the intermediate was substantially completely converted. Quenched to neutrality by adding sodium hydroxide solution at low temperature. After concentrating the methanol, the ethyl acetate was extracted, and the organic phase was dried over anhydrous sodium sulfate.
  • sodium borohydride temperature less than 20 deg.
  • 3-(3-bromophenyl)toluene (0.247 g) was dissolved in 3 mL of tetrahydrofuran, cooled to -78 ° C, butyl lithium (0.44 mL) was added dropwise to the reaction system, stirred for 30 min, then compound 94 ( 0.278g) of tetrahydrofuran solution was added dropwise to the reaction system, and reacted at -78 ° C for 1 h, and reacted at room temperature for 1 h. TLC detection of the raw material reaction was completely new, the saturated sodium chloride aqueous solution was quenched, ethyl acetate was extracted, and anhydrous sulfuric acid was obtained.
  • the crude compound 105 (60 mg) was dissolved in 5 mL of dichloromethane, and then, to the mixture of 2,6-dimethylpyridine, trifluoromethanesulfonic acid anhydride (2.0 eq.) was added dropwise. After 10 min of reaction, TLC showed the reaction was complete and quenched by water. After extraction with dichloromethane, it was dried and purified by column chromatography to yield 30 mg of white solid.
  • Example 15 of the preparation of the compound LW1005015 this example was hydrolyzed with the compound LW1005016, and purified to give the compound LW1005017.
  • MS-ESI [M+H] + 394.
  • Example 15 of the preparation of the compound LW1005015 this example was hydrolyzed with the compound LW1005018, and purified to give the compound LW1005019.
  • MS-ESI [M+H] + 394.
  • the m-bromo salicylaldehyde was dissolved in 10 mL of tetrahydrofuran, and 2 g of triethylamine and MOMCl (0.76 mL) were added under ice water. After 1 h, TLC showed the reaction was complete and quenched by water. The organic layer was washed with water and a saturated sodium chloride solution and dried over anhydrous sodium sulfate. The concentrated oil is directly subjected to the next reaction.
  • Test Example 1 Inhibitory effect of test compounds on mutual binding of PD-1/PD-L1 protein
  • HTRF homogeneous time-resolved fluorescence analysis
  • PD1/PDL1 binding assay kit (PD1/PDL1 binding assay kit-indirect format 1000tests, Cat. No.: 64CUS000C-1, Cisbio)
  • the compound was diluted in a gradient, and 2 ⁇ L of the compound or the dilution buffer was added to 4 ⁇ L of Tag1-PD-L1 protein (final concentration of 2 nM), and the mixture was preincubated for 15 minutes under normal temperature. Then 4 ⁇ L of Tag2-PD1 protein (final concentration 10 nM) was added and further incubated for 15 minutes. Subsequently, 5 ⁇ L of anti-Tag1-Eu 3+ antibody and 5 ⁇ L of anti-Tag2-XL665 antibody or 10 ⁇ L of a pre-mix of anti-Tag1-Eu 3+ antibody and anti-Tag2-XL665 antibody were added. The plate was sealed, and the reaction solution was reacted at room temperature for 2 hours, and the 665 nm and 620 nm fluorescent signals were detected on an EnVision plate reader.
  • the HTRF ratio was calculated from the fluorescence emission values of 665 nm and 620 nm.
  • IR inhibition rate
  • LW1005012 22.7 5 LW1005013 27.4 6 LW1005014 22.8 7 LW1005015 15.3 8 LW1005016 19.4 9 LW1005017 20.1 10 LW1005018 19.4 11 LW1005019 24.2 12 LW1005020 29.5 13 LW1005021 18.0 14 LW1005022 20.6

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明公开了一种PD-1/PD-L1小分子抑制剂及其制备方法和用途。具体地，本发明公开了结构如式L所示的化合物，其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物，具体定义详见说明书。还公开了该类化合物在抑制PD-1/PD-L1方面具有优异效果。

Description

PD-1/PD-L1小分子抑制剂及其制备方法和用途 技术领域
本发明属于药物合成领域，具体的说是本发明涉及PD-1/PD-L1的抑制剂及其制备方法和用途。
背景技术
程序性细胞死亡(programmed death-1 PD-1)是一种50-55KD的I型跨膜糖蛋白属于免疫球蛋白超家族成员，其显著的特点就是胞浆区分别含有N-端和C-端两个酪氨酸残基。前者参与构成一个免疫受体酪氨酸抑制基序(ITIM)，后者则参与构成一个免疫受体酪氨酸转换基序(ITSM)。PD-1与配体结合后，ITSM区的酪氨酸发生磷酸化，蛋白酪氨酸磷酸酶分子则被招募，使下游的效应分子去磷酸化转导负性信号从而发挥负性调节作用抑制T细胞的增殖和细胞因子的产生。
PD-1以单体形式存在于细胞表面，最早表达于胸腺中的双阴性细胞，也可表达于活化的T细胞、B细胞、自然杀伤细胞、树突状细胞以及活化的单核细胞。PD-1通常与其配体共同组成信号通路，其配体有PD-L1和PD-L2，这两种配体有40％的氨基酸序列相同。细胞外区有IgC和IgV型结构域，它们结构相似，但是分布不同。PD-L1分布广泛表达于鼠源T细胞，B细胞，树突状细胞，巨噬细胞和干细胞培养的骨髓细胞。虽然人和鼠PD-1的核苷酸序列具有70％的同源性，都编码一个288个氨基酸残基组成的蛋白质，并且氨基酸水平上有60％的同源性，但人源PD-L1的表达较鼠源的低。PD-L2分布相对比较局限主要表达在活化的单核巨噬细胞和树突状细胞。PD-1与其配体(主要是PD-L1)具有负性免疫调节作用。
在正常情况下，PD-1/PD-L1信号通路可以诱导和维持外周组织的免疫耐受，对防止组织的过度炎症反应以及自身免疫性疾病的发生具有积极作用。而在不正常的情况下，此信号通路在抗病毒或者抗肿瘤时也发挥着重要的调节作用。例如，PD-L1在肿瘤细胞以及免疫细胞中表达，PD-1与其结合减少了T细胞在癌症患者体内生成，抑制抗肿瘤免疫应答并导致T细胞耗尽。在病毒性疾病中，病毒使用类似的机制破坏有效的免疫识别。当前PD-1/PD-L1系列的药物主要是单克隆抗体，并且已经显示令人印象深刻的临床，适应症包括黑色素瘤和肺癌。
目前，两种PD-1抑制剂的单克隆抗体通过FDA批准上市，包括merck的pembrolizumab以及BMS的nivolumab。另外2016年，罗氏的PD-L1抑制剂Atezolizumab也通过FDA批准上市用于作为二线药物治疗晚期膀胱癌。然而，单克隆抗体同样表现出明显的缺点，例如非常有限的组织和肿瘤穿透，半衰期长，缺乏口服生物利用度以及困难和昂贵的生产。此外，当前靶向PD-1/PD-L1单克隆抗体仅在一小部分病例和肿瘤类型中有响应。因此，寻找非单克隆抗体，包括小分子，肽，环肽和大环化合物是很有必要的。截至目前，PD-1/PD-L1小分子抑制剂的报道仅有Bristol-Meyers Squibb的专利WO2015160641，发展新型的PD-1/PD-L1的小分子抑制剂是很有前景的。
发明内容
本发明的目的是提供一种新型的抑制PD-1/PD-L1的小分子以及其制备方法和用途。本文所述的化合物是能够抑制PD-1/PD-L1生物活性，具有治疗以及研究用途。
本发明的第一方面，提供了一种如式L所示的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物：
Figure PCTCN2018110869-appb-000001
其中，环A由环B和环C并环形成，如式L-1和式L-2所示：
Figure PCTCN2018110869-appb-000002
其中，
R 1、R 2、R 3、R 4各自独立地为氢、氧代(O或＝O)、卤素、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、-CN、羟基、氨基、羧基、取代或未取代的C6-C10芳基、取代或未取代的C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、卤代的具有1-3个选自N、S和O的杂原子的5-10元杂芳基，或-(CH 2) n-Ar；其中，n为0、1、2、3或4；Ar为未取代的或取代的C6-C10芳基、未取代的或取代的5-10元杂芳基、未取代的或取代的5-10元杂芳基；所述取代的是指被选自下组的一个或多个取代基所取代：羧基(-COOH)、酯基(C1-C6烷基O(CO)-)；或为无；
环B与环C各自独立地为5-6元芳香环、5-6元芳香杂环、5-6元饱和脂肪环或饱或5-6元脂肪杂环；且所述的环B和环C组成并环结构；
X为无(即R 5和环C或环B直接相连)、5-10元的芳基或杂芳基；
R 5为未取代的或取代的选自下组的基团：5-10元芳基、5-10元杂芳基、5-10元环基、5-10元杂环基；所述取代的是指被选自下组的一个或多个取代基所取代：C1-C6烷基、卤代C1-C6烷基、羧基(-COOH)、酯基(C1-C6烷基O(CO)-)；
所述杂环基、杂芳基、芳香杂环或饱和脂肪杂环包含1-3个(1个，2个或3个)选自N、S和O的杂原子；
除非特别说明，所述的取代指基团上的一个或多个氢原子被选自下组的取代基所取代：卤素、C1-C6烷基、卤代的C1-C6烷基、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、氧代、-CN、羟基、氨基、羧基、C6-C10芳基、卤代的C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、卤代的具有1-3 个选自N、S和O的杂原子的5-10元杂芳基。
在另一优选例中，R 5-X-为选自下组的结构：
Figure PCTCN2018110869-appb-000003
其中，R 10、R 11、R 12、R m、R n、R L各自独立地选自下组：H、卤素、C1-C6烷基、卤代的C1-C6烷基、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、氧代、-CN、羟基、氨基、羧基、C6-C10芳基、卤代的C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、卤代的具有1-3个选自N、S和O的杂原子的5-10元杂芳基。
在另一优选例中，式L所示的化合物选自下组：
Figure PCTCN2018110869-appb-000004
在另一优选例中，式L所示的化合物选自下组：
Figure PCTCN2018110869-appb-000005
Figure PCTCN2018110869-appb-000006
各式中，各基团的定义同前。
在另一优选例中，所述的化合物选自下组：
Figure PCTCN2018110869-appb-000007
本发明的第二方面，提供了如本发明第一方面所述化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物的制备方法：
(1)所述化合物为式I-5化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000008
或(2)所述化合物为式II-5化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000009
或(3)所述化合物为式III-5化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000010
或(4)所述化合物为式IV-7化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000011
或(5)所述化合物为式V-7化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000012
或(6)所述化合物为式VI-8化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000013
或(7)所述化合物为式VII-4化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000014
或(8)所述化合物为式VIII-8化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000015
或(9)所述化合物为式IX-7化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000016
或(10)所述化合物为式X-6化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000017
或(11)所述化合物为式XI-7化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000018
或(12)所述化合物为式XII-7化合物，所述方法包括步骤：
Figure PCTCN2018110869-appb-000019
各式中，各个基团的定义同前。
本发明的第三方面，提供了一种如下各式所示的中间体，其立体异构体、顺反异构体或互变异构体，
Figure PCTCN2018110869-appb-000020
Figure PCTCN2018110869-appb-000021
Figure PCTCN2018110869-appb-000022
各式中，R 1、R 2、R 3、R 4和R 5的定义同前。
本发明的第四方面，提供了一种药物组合物，包含(1)如本发明第一方面所述的化合物、其立体异构体或互变异构体、或其药学上可接受的盐、水合物或溶剂化物；(2)药学上可接受的载体。
本发明的第四方面，提供了如本发明第一方面所述的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物或如本发明第三方面所述药物组合物的用途，其特征在于，用于制备PD-1/PD-L1抑制剂。
本发明的第五方面，提供了如本发明第一方面所述的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物或如本发明第三他们所述药物组合物的用途，其特征在于，用于制备治疗癌症的药物；优选地，所述癌症选自下组：黑色素瘤、肺癌、膀胱癌。
本发明的第六方面，提供了如本发明第一方面所述的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物或如本发明第三他们所述药物组合物的用途，其特征在于，用于制备治疗或预防与PD-1/PD-L1活性或表达量相关的疾病的药物组合物。
本发明另一方面提供了一种治疗方法，所述方法包括步骤：给有需要的患者施用如本发明第一方面所述化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、 水合物或溶剂化物或如本发明第三方面所述药物组合物。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
本发明人通过广泛而深入的研究，首次意外地发现一种具有很好的生物活性的新型PD-1小分子抑制剂。在此基础上完成了本发明。
术语
除非另外定义，否则本文中所用的全部技术与科学术语均具有如本发明所属领域的普通技术人员通常理解的相同含义。
如本文所用，在提到具体列举的数值中使用时，术语“约”意指该值可以从列举的值变动不多于1％。例如，如本文所用，表述“约100”包括99和101和之间的全部值(例如，99.1、99.2、99.3、99.4等)。
如本文所用，术语“含有”或“包括(包含)”可以是开放式、半封闭式和封闭式的。换言之，所述术语也包括“基本上由…构成”、或“由…构成”。
定义
如本文所用，术语“烷基”包括直链或支链的烷基。例如C 1-C 8烷基表示具有1-8个碳原子的直链或支链的烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基等。
如本文所用，术语“烯基”包括直链或支链的烯基。例如C 2-C 6烯基指具有2-6个碳原子的直链或支链的烯基，例如乙烯基、烯丙基、1-丙烯基、异丙烯基、1-丁烯基、2-丁烯基、或类似基团。
如本文所用，术语“炔基”包括直链或支链的炔基。例如C 2-C 6炔基是指具有2-6个碳原子的直链或支链的炔基，例如乙炔基、丙炔基、丁炔基、或类似基团。
如本文所用，术语“C 3-C 8环烷基”指具有3-8个碳原子的环烷基。其可以是单环，例如环丙基、环丁基、环戊基、环己基、或类似基团。也可以是双环形式，例如桥环或螺环形式。
如本文所用，术语“C 1-C 8烷氧基”是指具有1-8个碳原子的直链或支链的烷氧基；例如，甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、异丁氧基、叔丁氧基等。
如本文所用，术语“具有1-3个选自下组N、S和O的杂原子的3-10元杂环烷基”是指具有3-10个原子的且其中1-3个原子为选自下组N、S和O的杂原子的饱和或部分饱和的环状基团。其可以是单环，也可以是双环形式，例如桥环或螺环形式。具体的实例可以为氧杂环丁烷、氮杂环丁烷、四氢-2H-吡喃基、哌啶基、四氢呋喃基、吗啉基和吡咯烷基等。
如本文所用，术语“C 6-C 10芳基”是指具有6-10个碳原子的芳基，例如，苯基或萘基 等类似基团。
如本文所用，术语“具有1-3个选自下组N、S和O的杂原子的5-10元杂芳基”指具有5-10个原子的且其中1-3个原子为选自下组N、S和O的杂原子的环状芳香基团。其可以是单环，也可以是稠环形式。具体的实例可以为吡啶基、哒嗪基、嘧啶基、吡嗪基、三嗪基、吡咯基、吡唑基、咪唑基、(1,2,3)-三唑基以及(1,2,4)-三唑基、四唑基、呋喃基、噻吩基、异恶唑基、噻唑基、恶唑基等。
除非特别说明，否则本发明的基团均可被选自下组的取代基所取代：卤素、腈基、硝基、羟基、氨基、C 1-C 6烷基-胺基、C 1-C 6烷基、C 2-C 6烯基、C 2-C 6炔基、C 1-C 6烷氧基、卤代C 1-C 6烷基、卤代C 2-C 6烯基、卤代C 2-C 6炔基、卤代C 1-C 6烷氧基、烯丙基、苄基、C 6-C 12芳基、C 1-C 6烷氧基-C 1-C 6烷基、C 1-C 6烷氧基-羰基、苯氧羰基、C 2-C 6炔基-羰基、C 2-C 6烯基-羰基、C 3-C 6环烷基-羰基、C 1-C 6烷基-磺酰基等。
如本文所用，“卤素”或“卤原子”指F、Cl、Br、和I。更佳地，卤素或卤原子选自F、Cl和Br。“卤代的”是指被选自F、Cl、Br、和I的原子所取代。
除非特别说明，本发明所描述的结构式意在包括所有的同分异构形式(如对映异构，非对映异构和几何异构体(或构象异构体))：例如含有不对称中心的R、S构型，双键的(Z)、(E)异构体等。因此，本发明化合物的单个立体化学异构体或其对映异构体、非对映异构体或几何异构体(或构象异构体)的混合物都属于本发明的范围。
如本文所用，术语“互变异构体”表示具有不同能量的结构同分异构体可以超过低能垒，从而互相转化。比如，质子互变异构体(即质子移变)包括通过质子迁移进行互变，如1H-吲唑与2H-吲唑。化合价互变异构体包括通过一些成键电子重组而进行互变。
如本文所用，术语“溶剂合物”是指本发明化合物与溶剂分子配位形成特定比例的配合物。
如本文所用，术语“水合物”是指本发明化合物与水进行配位形成的配合物。
活性成分
如本文所用，“本发明化合物”指式(L)所示的化合物，并且还包括及式(L)化合物的各种晶型形式、药学上可接受的盐、水合物或溶剂合物。
如本文所用，“药学上可接受的盐”指本发明化合物与酸或碱所形成的适合用作药物的盐。药学上可接受的盐包括无机盐和有机盐。一类优选的盐是本发明化合物与酸形成的盐。适合形成盐的酸包括但并不限于：盐酸、氢溴酸、氢氟酸、硫酸、硝酸、磷酸等无机酸，甲酸、乙酸、丙酸、草酸、丙二酸、琥珀酸、富马酸、马来酸、乳酸、苹果酸、酒石酸、柠檬酸、苦味酸、甲磺酸、苯甲磺酸，苯磺酸等有机酸；以及天冬氨酸、谷氨酸等酸性氨基酸。
本发明所制备得到的新型PD-1抑制剂，即式L化合物如下表1所示：
表1 本发明化合物列表
Figure PCTCN2018110869-appb-000023
Figure PCTCN2018110869-appb-000024
Figure PCTCN2018110869-appb-000025
Figure PCTCN2018110869-appb-000026
药物组合物和施用方法
由于本发明化合物具有优异的PD-1的抑制活性，因此本发明化合物及其各种晶型，药学上可接受的无机或有机盐，水合物或溶剂合物，以及含有本发明化合物为主要活性成分的药物组合物可用于预防和/或治疗与PD-1/PD-L1信号通路相关的疾病(例如，癌症)。
本发明的药物组合物包含安全有效量范围内的本发明化合物及药学上可以接受的赋形剂或载体。其中“安全有效量”指的是：化合物的量足以明显改善病情，而不至于产生严重的副作用。通常，药物组合物含有1-2000mg本发明化合物/剂，更佳地，含有10-200mg本发明化合物/剂。较佳地，所述的“一剂”为一个胶囊或药片。
“药学上可接受的载体”指的是：一种或多种相容性固体或液体填料或凝胶物质，它们适合于人使用，而且必须有足够的纯度和足够低的毒性。“相容性”在此指的是组合物中各组份 能和本发明化合物以及它们之间相互掺和，而不明显降低化合物的药效。药学上可以接受的载体部分例子有纤维素及其衍生物(如羧甲基纤维素钠、乙基纤维素钠、纤维素乙酸酯等)、明胶、滑石、固体润滑剂(如硬脂酸、硬脂酸镁)、硫酸钙、植物油(如豆油、芝麻油、花生油、橄榄油等)、多元醇(如丙二醇、甘油、甘露醇、山梨醇等)、乳化剂(如吐温
Figure PCTCN2018110869-appb-000027
)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。
本发明化合物或药物组合物的施用方式没有特别限制，代表性的施用方式包括(但并不限于)：口服、肠胃外(静脉内、肌肉内或皮下)。
用于口服给药的固体剂型包括胶囊剂、片剂、丸剂、散剂和颗粒剂。在这些固体剂型中，活性化合物与至少一种常规惰性赋形剂(或载体)混合，如柠檬酸钠或磷酸二钙，或与下述成分混合：(a)填料或增容剂，例如，淀粉、乳糖、蔗糖、葡萄糖、甘露醇和硅酸；(b)粘合剂，例如，羟甲基纤维素、藻酸盐、明胶、聚乙烯基吡咯烷酮、蔗糖和阿拉伯胶；(c)保湿剂，例如，甘油；(d)崩解剂，例如，琼脂、碳酸钙、马铃薯淀粉或木薯淀粉、藻酸、某些复合硅酸盐、和碳酸钠；(e)缓溶剂，例如石蜡；(f)吸收加速剂，例如，季胺化合物；(g)润湿剂，例如鲸蜡醇和单硬脂酸甘油酯；(h)吸附剂，例如，高岭土；和(i)润滑剂，例如，滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、十二烷基硫酸钠，或其混合物。胶囊剂、片剂和丸剂中，剂型也可包含缓冲剂。
固体剂型如片剂、糖丸、胶囊剂、丸剂和颗粒剂可采用包衣和壳材制备，如肠衣和其它本领域公知的材料。它们可包含不透明剂，并且，这种组合物中活性化合物或化合物的释放可以延迟的方式在消化道内的某一部分中释放。可采用的包埋组分的实例是聚合物质和蜡类物质。必要时，活性化合物也可与上述赋形剂中的一种或多种形成微胶囊形式。
用于口服给药的液体剂型包括药学上可接受的乳液、溶液、悬浮液、糖浆或酊剂。除了活性化合物外，液体剂型可包含本领域中常规采用的惰性稀释剂，如水或其它溶剂，增溶剂和乳化剂，例知，乙醇、异丙醇、碳酸乙酯、乙酸乙酯、丙二醇、1,3-丁二醇、二甲基甲酰胺以及油，特别是棉籽油、花生油、玉米胚油、橄榄油、蓖麻油和芝麻油或这些物质的混合物等。
除了这些惰性稀释剂外，组合物也可包含助剂，如润湿剂、乳化剂和悬浮剂、甜味剂、矫味剂和香料。
除了活性化合物外，悬浮液可包含悬浮剂，例如，乙氧基化异十八烷醇、聚氧乙烯山梨醇和脱水山梨醇酯、微晶纤维素、甲醇铝和琼脂或这些物质的混合物等。
用于肠胃外注射的组合物可包含生理上可接受的无菌含水或无水溶液、分散液、悬浮液或乳液，和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的含水和非水载体、稀释剂、溶剂或赋形剂包括水、乙醇、多元醇及其适宜的混合物。
本发明化合物可以单独给药，或者与其他药学上可接受的化合物联合给药。
联合给药时，所述药物组合物还包括与一种或多种其他药学上可接受的化合物。该其他药学上可接受的化合物中的一种或多种可与本发明的化合物同时、分开或顺序地给药。
使用药物组合物时，是将安全有效量的本发明化合物适用于需要治疗的哺乳动物(如人)，其中施用时剂量为药学上认为的有效给药剂量，对于60kg体重的人而言，日给药剂量通常为1～2000mg，优选20～500mg。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。
本发明的主要优点在于：
本发明的化合物结构新颖且具有优异的PD-1/PD-L1抑制作用。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数是重量百分比和重量份数。
通用材料及测试方法：
实施例中涉及到的仪器及原料说明如下：
核磁共振氢谱是Bruker AV-400(400MHz)核磁仪分析得到。化学位移以四甲基硅烷为内标来记录，以ppm为单位来表示(CDC1 3：δ7.26ppm)。记录的数据信息如下:化学位移及其裂分和偶合常数(s:单重峰；d:双重峰；t：三重峰；q：四重峰；br：宽峰；m：多重峰)。质谱数据采用菲尼根高级LCQ公司(Finnigan LCQAdvantage)的液质联用仪进行分析。
所有反应都在干燥氩气保护的无水无氧条件下进行操作。固体金属有机化合物在氩气保护干燥箱中进行储藏。四氢呋喃和乙醚是经过蒸馏得到，蒸馏时在其中加入金属钠和二苯甲酮。二氯甲烷，戊烷和己烷是用氢化钙来处理。
本发明所涉及的原料和试剂除特殊说明外均可市售或订制加工购买得到。
本发明中化合物可含有一个或多个不对称中心，因此该系列化合物可为消旋或者单一对映体形式。本发明所制备的化合物(通式IV)是纯度高于95％的杂环化合物，每个最终产物的结构表征分别由MS或/和氢谱核磁共振( 1HNMR)分析确定。以下通过实施例说明本发明各类化合物和中间体的合成。
实施例1：化合物LW1005001合成
Figure PCTCN2018110869-appb-000028
步骤:1-1：
Figure PCTCN2018110869-appb-000029
2-氨基-4-溴苯甲醛(200mg)，3-(3-甲基)苯乙酮(210mg)，氢氧化钾(112mg)依次加入到10mL无水乙醇中，氩气保护下回流2h。TLC显示原料基本反应完全，浓缩后加入 水以及乙酸乙酯萃取。有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到无色油状物277mg。 1H NMR(CDCl 3,400M):δ2.46(s,3H),7.21(d,J＝7.6Hz,1H),7.38(t,J＝7.6Hz,1H),7.49-7.52(m,2H),7.57-7.63(m,2H),7.69-7.71(m,2H),7.95(d,J＝8.8Hz,1H),8.11(d,J＝7.6Hz,1H),8.20(d,J＝8.4Hz,1H),8.38(d,J＝9.6Hz,1H).
步骤1-2：
Figure PCTCN2018110869-appb-000030
化合物3(270mg)，邻硼酸苯甲酸甲酯(194mg)，Pd(PPh 3) 4(42mg)，碳酸钾(199mg)加入到10mLDMF中，抽换氩气三次后加热到100℃反应5h。TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到黄色油状物化合物4 220mg。 1H NMR(CDCl 3,400M):δ2.45(s,3H),3.64(s,3H),7.20(d,J＝7.6Hz,1H),7.37(t,J＝7.2Hz,1H),7.46-7.53(m,5H),7.57-7.62(m,2H),7.69(d,J＝7.6Hz,1H),7.84(d,J＝8.8Hz,1H),7.94-7.97(m,2H),8.14(d,J＝8.0Hz,1H),8.17(s,1H),8.27(d,J＝8.4Hz,1H),8.39(s,1H).
步骤1-3：
Figure PCTCN2018110869-appb-000031
化合物4(60mg)溶于MeOH/THF(3mL/2mL)，加入氢氧化钠溶液(2mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到淡黄色固体25mg。 1H NMR(CDCl 3,400M):δ2.42(s,3H),6.35(br,1H),7.18(d,J＝7.6Hz,1H),7.33(t,J＝7.2Hz,1H),7.39-7.47(m,5H),7.50-7.57(m,2H),7.64(d,J＝7.2Hz,1H),7.70(d,J＝8.4Hz,1H),7.77(d,J＝8.0Hz,1H),7.92-7.97(m,2H),8.14(d,J＝8.8Hz,1H),8.23(d,J＝8.0Hz,2H).MS-ESI:[M-H] +＝416.
实施例2：化合物LW1005002合成
Figure PCTCN2018110869-appb-000032
步骤2-1：
Figure PCTCN2018110869-appb-000033
化合物11(50mg)溶于5mL二氯甲烷中，加入间氯过氧苯甲酸40mg(2.0eq.)，室温 反应20h，饱和碳酸氢钠溶液淬灭反应。二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥。浓缩后柱层析得到棕色固体25mg。 1H NMR(CDCl 3,400M):δ2.43(s,3H),3.67(s,3H),7.18(d,J＝7.2Hz,1H),7.34(t,J＝7.6Hz,1H),7.45-7.57(m,4H),7.58-7.61(m,4H),7.69(d,J＝7.6Hz,1H),7.80(d,J＝8.4Hz,1H),7.88(d,J＝8.8Hz,1H),7.94-7.97(m,2H),8.20(s,1H),8.85(s,1H).
步骤2-2：
Figure PCTCN2018110869-appb-000034
化合物12(20mg)溶于MeOH/THF(3mL/2mL)，加入氢氧化钠溶液(2mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到黄色固体8mg， 1H NMR(d6-DMSO,400M):δ2.40(s,3H),3.67(s,3H),7.22(d,J＝7.6Hz,1H),7.39(t,J＝8.0Hz,1H),7.53-7.59(m,4H),7.61-7.72(m,4H),7.79(d,J＝7.6Hz,1H),7.87-7.89(m,2H),8.00(d,J＝7.6Hz,1H),8.00(d,J＝8.4Hz,1H),8.15(d,J＝8.0Hz,1H),8.27(s,1H),8.57(s,1H).MS-ESI:[M-H] +＝432.
实施例3：化合物LW1005003的合成
Figure PCTCN2018110869-appb-000035
步骤3-1：
Figure PCTCN2018110869-appb-000036
底物(1g)溶于30mL氯仿中加热到45℃，缓慢滴加液溴(0.83g)。滴加完毕后TLC显示反应完全。降温后硫代硫酸钠溶液洗涤，有机相以水，饱和氯化钠溶液洗涤后，无水硫酸钠干燥。浓缩后柱层析得到淡黄色色固体0.9g。 1H NMR(CDCl 3,400M):δ2.37(s,3H),4.43(s,2H),7.15(d,J＝7.2Hz,1H),7.28-7.35(m,3H),7.49(t,J＝7.2Hz,1H),7.75-7.77(m,1H),7.87-7.89(m,1H),8.12(t,J＝2.0Hz,1H).
步骤3-2：
Figure PCTCN2018110869-appb-000037
化合物22(500mg)加入到10mL DMSO中，随后加入碳酸氢钠175mg以及4-溴苯二胺390mg，氩气保护下加热到120℃，反应4h。TLC显示反应完全，生成两个产物。降温后加入水洗涤，乙酸乙酯萃取。有机相以饱和食盐水洗涤，无水硫酸钠干燥。浓缩得到化合物23(155mg)。： 1H NMR(CDCl 3,400M):δ2.47(s,3H),7.23(d,J＝7.6Hz,1H),7.39(t,J＝8.0Hz,1H),7.49-7.51(m,2H),7.64(t,J＝8.0Hz,1H),7.74-7.77(m,1H),7.87(dd,J＝8.8,2.4Hz,1H),8.05(d,J＝8.8Hz,1H),8.14(td,J＝7.6,1.2Hz,1H),8.32(d,J＝2.8Hz,1H),8.40(t,J＝1.6Hz,1H),9.38(s,1H).
步骤3-3：
Figure PCTCN2018110869-appb-000038
化合物23(150mg),邻硼酸苯甲酸甲酯(108mg),Pd(PPh 3) 4(24mg),碳酸钾(110mg)加入到5mLDMF中，抽换氩气三次后加热到100℃反应16h。TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到白色固体100mg。 1H NMR(CDCl 3,400M):δ2.47(s,3H),3.66(s,3H),7.23(d,J＝7.6Hz,1H),7.40(t,J＝7.6Hz,1H),7.50-7.53(m,4H),7.61-7.67(m,2H),7.73-7.77(m,2H),7.98(d,J＝7.6Hz,1H),8.09(s,1H),8.16-8.20(m,2H),8.43(s,1H),9.41(s,1H).
步骤3-4：
Figure PCTCN2018110869-appb-000039
化合物24(100mg)溶于MeOH/THF(6mL/4mL),加入氢氧化钠溶液(4mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体70mg。 1H NMR(CDCl 3,400M):δ2.45(s,3H),7.21(d,J＝7.2Hz,1H),7.37(t,J＝7.6Hz,1H),7.48-7.53(m,4H),7.59-7.66(m,2H),7.73(d,J＝8.0Hz,2H),7.85(dd,J＝8.8,1.6Hz,1H),8.02-8.04(m,1H),8.12(d,J＝8.0Hz,1H),8.19-8.21(m,2H),8.37(s,1H),9.33(s,1H).MS-ESI:[M-H] +＝417.
实施例4：化合物LW1005004的合成
Figure PCTCN2018110869-appb-000040
步骤4-1：
Figure PCTCN2018110869-appb-000041
化合物31(500mg)加入到10mL DMSO中，随后加入碳酸氢钠175mg以及4-溴苯二胺390mg，氩气保护下加热到120℃，反应4h。TLC显示反应完全，生成两个产物。降温后加入水洗涤，乙酸乙酯萃取。有机相以饱和食盐水洗涤，无水硫酸钠干燥。浓缩得到化合物(260mg) 1H NMR(CDCl 3,400M):δ2.47(s,3H),7.23(d,J＝8.0Hz,1H),7.39(t,J＝7.6Hz,1H),7.49-7.51(m,2H),7.64(t,J＝8.0Hz,1H),7.75-7.78(m,1H),7.83(dd,J＝8.8,2.4Hz,1H),8.01(d,J＝8.8Hz,1H),8.13-8.16(m,1H),8.32(d,J＝2.8Hz,1H),8.38-8.42(m,1H),9.39(s,1H).
步骤4-2：
Figure PCTCN2018110869-appb-000042
化合物32(100mg),邻硼酸苯甲酸甲酯(72mg),Pd(PPh 3) 4(16mg),碳酸钾(74mg)加入到5mLDMF中，抽换氩气三次后加热到100℃反应16h。TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到白色固体70mg。 1H NMR(CDCl 3,400M):δ2.46(s,3H),3.66(s,3H),7.22(d,J＝8.0Hz,1H),7.39(t,J＝7.6Hz,1H),7.51-7.53(m,4H),7.61-7.64(m,2H),7.70(dd,J＝8.4,2.0Hz,2H),7.75(d.J＝8.0Hz,1H),7.98-8.00(m,1H),8.13-8.17(m,2H),8.43(s,1H),9.40(s,1H).
步骤4-3：
Figure PCTCN2018110869-appb-000043
化合物33(70mg)溶于MeOH/THF(6mL/4mL),加入氢氧化钠溶液(4mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体50mg。 1H NMR(CDCl 3,400M):δ2.46(s,3H),7.22(d,J＝8.4Hz,1H),7.38(t,J＝7.6Hz,1H),7.49-7.54(m,4H), 7.63(q,J＝7.6Hz,2H),7.70-7.75(m,2H),8.05-8.07(m,2H),8.14(d,J＝7.6Hz,1H),8.17(d,J＝1.2Hz,2H),8.41(s,1H),9.38(s,1H).MS-ESI:[M-H] +＝417.
实施例5：化合物LW1005005合成
Figure PCTCN2018110869-appb-000044
步骤5-1：
Figure PCTCN2018110869-appb-000045
0℃下往CBr 4(1.5eq.)的二氯甲烷溶液中加入三苯基膦(3.0eq.),搅拌反应15min，加入醛的二氯甲烷溶液，随后继续反应0.5h。将二氯甲烷浓缩后过滤，浓缩后柱层析，得到二溴化合物。0℃下向二溴化合物的THF溶液中滴加正丁基锂(2.5M，己烷)，滴加完毕后维持该温度继续搅拌0.5h。随后升温至室温反应1.5h，TLC检测显示反应完全。滴加水淬灭反应。乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到淡黄色油状物43。 1H NMR(CDCl 3,400M):δ2.42(s,3H),3.10(s,1H),7.19(d,J＝7.2Hz,1H),7.32-7.40(m,4H),7.46(dd,J＝7.6,1.2Hz,1H),7.55-7.58(m,1H),7.72(d,J＝1.2Hz,1H).
步骤5-2：
Figure PCTCN2018110869-appb-000046
化合物43(0.76g),2,6-二氯-3-溴吡啶(1g)以及Pd(PPh 3) 2Cl 2(0.18g)，CuI(0.05g),三乙胺(1.34g)溶于10mL DMF中，氮气保护下，60℃反应的尝试5h，原料反应完全。加水20mL，乙酸乙酯萃取，无水硫酸钠干燥，将有机相浓缩后柱层析得黄色油状物(0.15g,收率:10％)。 1HNMR(CDCl 3,400MHz):δ2.43(s,2H),7.19(d,J＝7.6Hz,1H),7.28(d,J＝8.0Hz,1H),7.33-7.46(m,4H),7.53(d,J＝7.6Hz,1H),7.61(d,J＝8.0Hz,1H),7.77-7.93(m,1H)。
步骤5-3：
Figure PCTCN2018110869-appb-000047
化合物44(0.15g)溶于10mL DMF中，然后将硫化钠(0.11g)加入反应体系，130℃反应的尝试3.5h，原料反应完全，有新点生成。加水10mL，乙酸乙酯萃取，无水硫酸钠干燥，将有机相浓缩后柱层析得黄色固体(0.08g，收率:53％)。 1HNMR(CDCl 3,400MHz):δ2.45(s,3H),7.22(d,J＝7.2Hz,1H),7.31(d,J＝8.4Hz,1H),7.35-7.39(m,1H),7.43-7.49(m,2H),7.50-7.53(m,2H),7.59(d,J＝7.6Hz,1H),7.66(d,J＝7.6Hz,1H),7.88(t,J＝1.6Hz,1H),7.96(d,J＝8.4Hz,1H)。
步骤5-4：
Figure PCTCN2018110869-appb-000048
化合物45(0.04g),邻甲酸苯硼酸(0.03g),四三苯基膦钯(14mg)，碳酸铯(77mg)溶于1.5mL DMF中，然后氮气保护下，100℃反应的尝试5h，原料反应完全，有新点生成。加水10mL，乙酸乙酯萃取，无水硫酸钠干燥，将有机浓缩后柱层析得黄色固体(0.012g，收率:24％)。 1HNMR(CDCl 3,400MHz):δ2.41(s,3H),7.24(d,J＝7.2Hz,1H),7.38-7.42(m,1H),7.54-7.79(m,10H),8.06(d,J＝5.2Hz,2H),8.28(d,J＝8.4Hz,1H).
实施例6：化合物LW1005006的合成
Figure PCTCN2018110869-appb-000049
步骤6-1：
Figure PCTCN2018110869-appb-000050
底物酮(0.68g)溶于THF中，加入NaH(26mg)以及二氯哌啶(0.48g)，氩气保护下加热回流5h，不加0.2gNaH回流过夜。降温后加入水淬灭反应，乙酸乙酯萃取，有机相无水硫酸钠干燥。浓缩后将粗品溶解到DMF中，加入碳酸钾0.85g，加热到100℃反应16h，降温后将DMF减压浓缩，加入水以及乙酸乙酯萃取，有机相以无水硫酸钠干燥，浓缩后逐层析，得到固体440mg。 1H NMR(CDCl 3,400M):δ2.38(s,3H),7.16(d,J＝7.6Hz,1H),7.23(s,1H),7.18(t,J＝7.2Hz,1H),7.39-7.42(m,2H),7.1(t,J＝8.0Hz,1H),7.61-7.64(m,1H),7.8-7.87(m,1H),8.11-8.12(m,1H),8.17(d,J＝2.4Hz,1H).MS-ESI:[M+H] +＝287.
步骤6-2：
Figure PCTCN2018110869-appb-000051
化合物53(150mg)溶于5mL乙酸中，加入钨酸钠以及双氧水，加热到45℃反应过夜。硫代硫酸钠溶液淬灭反应，乙酸乙酯萃取，有机相无水硫酸钠干燥。浓缩后柱层析得到白色固体100mg。
上步反应得到的白色固体54溶解到二氯甲烷中，氩气保护下加入三氯氧磷以及催化量的DMF，反应过夜后，TLC显示底物反应完全。加入水淬灭反应，二氯甲烷萃取后，有机相以无水硫酸钠干燥。浓缩后柱层析得到固体60mg。 1H NMR(CDCl 3,400M):δ2.47(s,3H),7.22-7.2(m,2H),7.39(t,J＝7.2Hz,1H),7.4-7.47(m,2H),7.9(t,J＝8.0Hz,1H),7.71(d,J＝8.0Hz,1H),7.91(d,J＝7.2Hz,1H),8.16(s,1H),8.23(s,1h).
步骤6-3：
Figure PCTCN2018110869-appb-000052
化合物55(44mg),邻硼酸苯甲酸甲酯(20mg),Pd(PPh 3) 4(5mg)，碳酸钾(45mg)加入到5mL DMF中，抽换氩气三次后加热到100℃反应5h。TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到淡黄色固体40mg。 1H NMR(CDCl 3,400M):δ2.40(s,3H),3.63(s,3H),7.13-7.18(m,2H),7.23(s,1H),7.31-7.7(m,7H),7.-7.7(m,2H),7.63(d,J＝8.0Hz,1H),7.87(t,J＝.6Hz,1H),8.13(s,1H),8.30(s,1H).
步骤6-4：
Figure PCTCN2018110869-appb-000053
化合物56(20mg)溶于MeOH/THF(5mL/2mL),加入氢氧化钠溶液(2mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到淡黄色固体9mg。 1H NMR(d6-DMSO,400M):δ2.43(s,3H),3.33(br,1H),7.25(d,J＝7.6Hz,1H),7.41(t,J＝8.0Hz,1H),7.59-7.70(m,6H),7.82(d,J＝8.0Hz,1H),7.88(d,J＝7.6Hz,1H),7.98(s,1H),8.06(d,J＝8.0Hz,1H),8.32(s,1H),8.49(s,1H).MS-ESI:[M+H] +＝407.
实施例7化合物LW1005007的合成
Figure PCTCN2018110869-appb-000054
步骤7-1：
Figure PCTCN2018110869-appb-000055
氩气保护下，3-甲氧基苯硫酚(6g)，以及2-溴-1,1-二乙氧基乙烷(12.75g)溶解到50mL丙酮中加入11.9g碳酸钾，加热回流4h，TLC显示苯硫酚反应完全，浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到无色油状物6g。
中间体(5g)溶解到50mL氯苯中，加入多聚磷酸5g，加热到110℃反应过夜，TLC显示反应完全。降温后滴加10％氢氧化钠溶液调节pH至7，加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体2.43g。 1H NMR(CDCl 3,400M):δ3.87(s,3H),6.98-7.01(m,1H),7.23-7.26(m,2H),7.34(d,J＝1.6Hz,1H),7.69(d,J＝9.2Hz,1H).
步骤7-2：
Figure PCTCN2018110869-appb-000056
干燥的两口瓶中加入化合物64(260mg)，THF(10mL)，降温至-70℃后滴加丁基锂(2mL，3eq),反应液呈红色。该温度下搅拌0.5h后滴加片哪醇硼酸甲酯(1.26g,5eq),将温度升至室温搅拌1h。加水淬灭反应，乙酸乙酯萃取，干燥后浓缩，粗品直接进行下步反应。MS-ESI:[M+H] +＝291.11
步骤7-3：
Figure PCTCN2018110869-appb-000057
化合物65(粗品)以及溴代物(394mg)溶解到无水DMF中，加入Pd(PPh 3) 4(92mg)以及碳酸钾(441mg)。抽换氩气后100℃反应16h，TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到白色固体180mg。 1H NMR(CDCl 3,400M):δ2.45(s,3H),3.89(s,3H),6.99(dd,J＝8.8,2.4Hz,1H),7.21(d,J＝7.2Hz,1H),7.32(d,J＝2.4Hz,1H),7.37(t,J＝7.2Hz,1H),7.44-7.49(m,3H),7.52-7.54(m,2H),7.63-7.68(m,2H),7.87(t,J＝2.0Hz,1H).
步骤7-4：
Figure PCTCN2018110869-appb-000058
化合物67(180mg)溶解到10mL二氯甲烷中，降温至0℃，滴加三溴化硼(409mg)，回复室温反应2h，TLC显示反应完全，加入水淬灭反应，乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到黄绿色固体150mg。 1H NMR(CDCl 3,400M):δ2.45(s,3H),6.91(d,J＝8.4Hz,1H),7.21(d,J＝7.6Hz,1H),7.28(s,1H),7.37(t,J＝7.2Hz,1H),7.44-7.53(m,3H),7.64(d,J＝8.0Hz,2H),7.86(d,J＝1.6Hz,1H).
步骤7-5：
Figure PCTCN2018110869-appb-000059
化合物67(150mg)溶解到10mL二氯甲烷中，加入三乙胺(142mg)后滴加三氟甲磺酸酐(268mg，2.0eq.)。室温反应2h后TLC显示反应完全，加入水淬灭反应。二氯甲烷萃取后干燥，柱层析得到白色固体170mg。 1H NMR(CDCl 3,400M):δ2.46(s,3H),7.22(d,J ＝7.6Hz,1H),7.26-7.30(m,1H),7.38(t,J＝7.2Hz,1H),7.44-7.46(m,2H),7.49-7.53(m,1H),7.59-7.62(m,2H),7.66(d,J＝7.6Hz,1H),7.77(s,1H),7.82(d,J＝8.8Hz,1H),7.89(s,1H).
步骤7-6：
Figure PCTCN2018110869-appb-000060
化合物68(170mg),邻硼酸苯甲酸甲酯(82mg),Pd(PPh 3) 4(22mg),碳酸钾(105mg)加入到10mLDMF中，抽换氩气三次后加热到100℃反应16h。TLC显示底物有少量剩余，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到黄色油状物54mg。 1H NMR(CDCl 3,400M):δ2.46(s,3H),3.65(s,3H),7.22(d,J＝7.6Hz,1H),7.31(d,J＝8.4Hz,1H),7.38(t,J＝7.2Hz,1H),7.42-7.47(m,4H),7.51(d,J＝8.0Hz,1H),7.70(d,J＝7.6Hz,1H),7.79-7.80(m,2H),7.86(d,J＝8.0Hz,1H),7.92(s,1H).
步骤7-7：
Figure PCTCN2018110869-appb-000061
化合物5-6(50mg)溶于MeOH/THF(6mL/4mL),加入氢氧化钠溶液(4mL,10％)，60℃反应2h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体26mg。 1H NMR(CDCl 3,400M):δ2.45(s,3H),7.21(d,J＝7.6Hz,1H),7.32(d,J＝8.0Hz,1H),7.37(t,J＝6.8Hz,1H),7.41-7.49(m,5H),7.54-7.60(m,3H),7.67(d,J＝7.6Hz,1H),7.76(d,J＝8.0Hz,1H),7.80(s,1H),7.90(s,1H),7.96(d,J＝7.6Hz,1H).
实施例8化合物LW1005008的合成
Figure PCTCN2018110869-appb-000062
步骤8-1：
Figure PCTCN2018110869-appb-000063
氩气保护下，化合物LW1005007(145mg)加入到10mL乙腈中，加入双氧水(4mL，30％水溶液)以及三氟乙酸酐(1.5mL)，室温搅拌16h。TLC显示反应完全，缓慢滴加饱和碳酸氢钠溶液淬灭反应。乙酸乙酯萃取，有机相以无水硫酸钠干燥。浓缩后柱层析得到黄色固体。(化合物溶解性差) 1H NMR(CDCl 3,400M):δ2.45(s,3H),7.21(d,J＝7.2Hz,1H),7.34-7.39(m,3H),7.41-7.46(m,3H),7.51-7.55(m,3H),7.60-7.76(m,2H),7.75(s,1H),7.82(d,J＝8.0Hz,1H),8.00(s,1H),8.06(d,J＝8.0Hz,1H).MS-ESI:[M+Na] +＝475.
步骤8-2：
Figure PCTCN2018110869-appb-000064
化合物72(75mg)加入到EtOH(10mL)中，随后加入THF(5mL)至底物溶解，加入Pd/C(10mg)，抽换氢气三次后反应过夜。过滤后浓缩，柱层析得到白色固体30mg。 1H NMR(CDCl 3,400M):δ2.42(s,3H),3.71(s,J＝7.6H,2H),4.79(t,J＝8.8H,1H),7.18(d,J＝7.6Hz,1H),7.31-7.41(m,4H),7.44-7.51(m,4H),7.59(d,J＝8.0Hz,1H),7.63-7.65(m,1H),7.69(s,1H),7.77(s,1H),8.02(d,J＝8.0Hz,1H).
实施例9：化合物LW1005009的合成
Figure PCTCN2018110869-appb-000065
步骤9-1：
Figure PCTCN2018110869-appb-000066
加入邻苯二酚(20g)至盐酸/1,4-二氧六环溶液(约9M)，搅拌溶清，再加入除水后的氯化锌，搅拌，TLC(取少量反应液，加水，加热到80度，冷却，加甲醇溶清，PE/EA＝2)显示反应完全(碘显色)；滴加水1200ml，加完后析出很多黄色固体，加热反应液到80度，保温搅拌约1h(TLC显示中间体完全转化)，冷却反应液到40度左右，将反应液倒入到冷水中(6L)，搅拌约2h，过滤出固体，甲苯带水后干燥得到黄色固体16g。 1H  NMR(d 6-DMSO,400M):δ5.01(s,2H),6.32(s,1H),6.38(dd,J＝8.8,2.0Hz,1H),7.72(d,J＝8.8Hz,1H),10.68(s,1H),11.65(s,1H).
步骤9-2：
Figure PCTCN2018110869-appb-000067
化合物81溶于甲醇中；冰浴下分批加入固体甲醇钠，控制温度低于20度，加完后TLC(取少量反应液加盐酸到酸性PE/EA＝2)显示反应完全，冰水浴，滴加2N HCl(3.5L)，温度低于20度，冷却搅拌至0～5度，搅拌1/2h，过滤出固体，甲苯带水后干燥得到红色固体10g。 1H NMR(d 6-DMSO,400M):δ4.69(s,2H),6.58(s,1H),6.63(dd,J＝8.0,2.0Hz,1H),7.43(d,J＝8.0Hz,1H),11.27(s,1H).
步骤9-3：
Figure PCTCN2018110869-appb-000068
化合物82加入到二氯甲烷中，不能完全溶解，冰水浴下加入咪唑。随后分批加TBSCl，温度20度以内，5min后TLC显示完全，加入水淬灭反应后二氯甲烷萃取后干燥。浓缩后的产物直接进行下步反应。
将粗品溶于甲醇，冰水浴下分批加入硼氢化钠(温度低于20度)，加完后搅拌1/2h，TLC显示反应完全。加入丙酮淬灭多余的硼氢化钠。随后加入四氢呋喃以及6N盐酸溶液至pH<1。将反应液加热到40℃反应0.5h，中间体基本完全转化。低温下加入氢氧化钠溶液淬灭至中性。将甲醇浓缩后乙酸乙酯萃取，有机相以无水硫酸钠干燥，柱层析得白色固体化合物84。 1H NMR(CDCl 3,400M):δ5.14(s,1H),6.69(s,1H),6.79(dd,J＝8.0,2.0Hz,1H),7.00(s,1H),7.42(d,J＝8.0Hz,1H),7.52(s,1H).
步骤9-4：
Figure PCTCN2018110869-appb-000069
化合物84(1g)溶于吡啶中加入三苯基氯甲烷(2.0eq)以及DMAP，加热到80℃反应3h，原料少量剩余。补加三苯基氯甲烷后原料仍然有剩余，停止反应，浓缩后加入水洗涤，乙酸乙酯，无水硫酸钠干燥。柱层析得到白色固体1g。 1H NMR(CDCl 3,400M):δ6.56(dd,J＝6.0,0.8Hz,1H),6.70(dd,J＝8.4,0.8Hz,1H),6.78(d,J＝1.6Hz,1H),7.17-7.28(m,10H),7.40(d,J＝2.0Hz,1H),7.46-7.49(m,6H).
步骤9-5：
Figure PCTCN2018110869-appb-000070
干燥的两口瓶中加入化合物85(200mg),THF(5mL)，降温至-70℃后滴加丁基锂(3eq),反应液呈红色。该温度下搅拌0.5h后滴加片哪醇硼酸甲酯(4eq),将温度升至室温搅拌0.5h。加水淬灭反应，乙酸乙酯萃取，干燥后浓缩，粗品直接进行下步反应。
步骤9-6：
Figure PCTCN2018110869-appb-000071
3-(3-溴苯基)甲苯,化合物86，Pd(PPh 3) 4,K 2CO 3加入到10mL DMF中，抽换氩气三次后加热到100℃反应16h。TLC显示反应完全，回复室温。将溶剂浓缩后，加入水以及乙酸乙酯萃取，有机相浓缩。浓缩液加入10mL甲醇进行溶解，滴入氯化氢/二氧六环溶液，10min后，调节pH＝5-6。将溶剂浓缩，加入水以及乙酸乙酯萃取，有机相浓缩后柱层析,得到黄色固体。 1H NMR(CDCl 3,400M):δ2.39(s,3H),6.72(dd,J＝8.0,2.0Hz,1H),6.94(d,J＝0.8Hz,1H),6.96(d,J＝0.8Hz,1H),7.09-7.15(m,3H),7.30-7.46(m,4H),7.71(m,1H),7.9(t,J＝1.2Hz,1H).
步骤9-7：
Figure PCTCN2018110869-appb-000072
化合物88溶解二氯甲烷中，加入三乙胺。氩气保护下滴加三氟甲磺酸酐1.5equiv，0.5h后，TLC显示反应完全。加入水淬灭反应，二氯甲烷萃取，浓缩后柱层析得到黄色固体。
步骤9-8：
Figure PCTCN2018110869-appb-000073
化合物89,邻硼酸苯甲酸甲酯,Pd(PPh 3) 4,碳酸钾加入到5mL DMF中，抽换氩气三次后加热到100℃反应5h。TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到淡黄色固体。 1H NMR(CDCl 3,400M):δ2.46(s,3H),3.65(s,3H),7.1(d,J＝1.2Hz,1H),7.19-7.21(m,2H),7.36-7.61(m,10H),7.83-7.86(m,2H),8.09(t,J＝1.6Hz,1H).
步骤9-9：
Figure PCTCN2018110869-appb-000074
化合物90(20mg)溶于MeOH/THF(5mL/2mL),加入氢氧化钠溶液(2mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体14mg。 1H NMR(CDCl 3,400M):δ2.45(s,3H),7.08(s,1H),7.17-7.23(m,2H),7.35-7.58(m,10H),7.81-7.84(m,1H),7.93(d,J＝7.6Hz,1H),8.07(t,J＝1.2Hz,1H).MS-ESI:[M+Na] +＝427.
实施例10：化合物LW1005010合成
Figure PCTCN2018110869-appb-000075
步骤10-1：
Figure PCTCN2018110869-appb-000076
将底物2,4-二溴苯乙酸甲酯(1.54g)，硼氢化钠(0.76g)，氯化钙(0.278g)溶于15mL THF中，然后加入5mL甲醇，常温下反应6h，TLC检测原料反应完全有新点生成，用0.1N的稀盐酸溶液淬灭反应，乙酸乙酯萃取，无水硫酸钠干燥，将有机相浓缩后得无色油状物(1.3g，收率：95％) 1HNMR(CDCl 3,400MHz):δ2.85-3.00(m,2H),3.85-3.89(m,3H),7.02-7.17(m,1H),7.31-7.39(m,2H),7.70(d,J＝6.0Hz,1H)。
步骤10-2：
Figure PCTCN2018110869-appb-000077
将化合物93(0.28g)，Dess-Martin氧化剂(0.63g)溶于6mL乙腈中，然后加热至80℃，TLC检测原料反应完全有新产物生成，用DCM稀释后，用饱和碳酸氢钠和硫代硫酸钠淬灭反应，DCM萃取，无水硫酸钠干燥，将有机相浓缩后柱层析得黄色粘稠状物(0.2g，收率：87％) 1HNMR(CDCl 3,400MHz):δ3.85-3.87(m,2H),7.11-7.45(m,1H), 7.45-7.49(m,1H),7.79-7.81(m,1H),9.76(s,1H)。
步骤10-3：
Figure PCTCN2018110869-appb-000078
3-(3-溴苯基)甲苯(0.247g)溶于3mL四氢呋喃中，降温至-78℃，将丁基锂(0.44mL)滴加入反应体系，滴加完毕搅拌30min，然后将化合物94(0.278g)的四氢呋喃溶液滴加入反应体系，-78℃下反应1h，常温下反应1h，TLC检测原料反应完全有新点生成，饱和氯化钠水溶液淬灭反应，乙酸乙酯萃取，无水硫酸钠干燥，将有机相浓缩后得无色油状物(0.1g，收率：22.4％) 1HNMR(CDCl 3,400MHz):δ2.44(s,3H),3.10-3.21(m,2H),5.04-5.07(m,1H),7.07(d,J＝8.0Hz,1H),7.18-7.19(m,1H),7.34-7.44(m,6H),7.51-7.55(m,2H),7.75(s,1H)。
步骤10-4：
Figure PCTCN2018110869-appb-000079
将化合物95(0.24g)溶于4mL甲苯中，然后将NaH(0.032g)，CuCl(2.5mg)加入反应体系，加热回流8h，TLC检测原料反应完全有新点生成，饱和氯化钠水溶液淬灭反应，乙酸乙酯萃取，无水硫酸钠干燥，将有机相浓缩后得到无色油状物(0.15g，收率：76％)。
步骤10-5：
Figure PCTCN2018110869-appb-000080
化合物13-4，邻羧基苯硼酸，碳酸铯，四(三苯基磷)钯溶于DMF中，氮气保护下100℃反应8h，TLC检测原料反应完全有新点生成，将水(20mL)加入反应体系，乙酸乙酯萃取，无水硫酸钠干燥，将有机相浓缩后柱层析得到化合物(5.0mg)。 1HNMR(CDCl 3,400MHz):δ2.44(s,3H),3.10-3.21(m,2H),5.04-5.07(m,1H),7.07(d,J＝8.0Hz,1H),7.18-7.19(m,1H),7.34-7.44(m,6H),7.51-7.55(m,2H),7.75(s,1H)。
实施例11：化合物LW1005011的合成
Figure PCTCN2018110869-appb-000081
步骤11-1：
Figure PCTCN2018110869-appb-000082
250mL单口烧瓶中加入硫酸溶液(46g，浓度80％)，剧烈搅拌下缓慢加入2,5-二甲基胺(30g),碘化钾(0.18g)以及甘油9.2g。混合液加热到外温145℃剧烈回流6h。反应后将反应液在冰浴下滴加氢氧化钠溶液调节pH>7。加入二氯甲烷萃取，合并有机相后以无水硫酸钠干燥。浓缩后减压蒸馏(或者柱层析)得到无色液体25g。 1H NMR(CDCl 3,400M):δ2.47(s,3H),2.63(s,3H),7.16(d,J＝6.8Hz,1H),7.34(d,J＝6.8Hz,1H),7.39-7.43(m,1H),8.22(dd,J＝8.4,1.6Hz,1H),8.86(d,J＝4.4Hz,1H).
步骤11-2：
Figure PCTCN2018110869-appb-000083
化合物102(10g)溶解到250mL乙腈中，碘单质(32g)溶解到反应液中，随后加入过氧化氢叔丁基(24g)。反应回流48h，其中补加两次过氧化氢叔丁基(16g)。底物仍然有剩余，降温后加入硫代硫酸钠溶液洗涤。乙酸乙酯萃取，有机相干燥后浓缩，快速柱层析得到103(800mg)。 1H NMR(CDCl 3,400M):δ2.52(s,3H),2.65(s,3H),7.18(d,J＝6.8Hz,1H),7.37(d,J＝6.8Hz,1H),8.56(s,1H),8.96(s,1H).
步骤11-3
Figure PCTCN2018110869-appb-000084
化合物103以及化合物103-1溶解到无水二氧六环中，加入Pd(PPh 3) 2Cl 2以及碳酸铯。抽换氩气后40℃反应16h，TLC显示底物完全转化。直接柱层析得到黄色固体80mg。 1H NMR(CDCl 3,400M):δ2.63(s,3H),2.69(s,3H),6.87(dd,J＝8.4,2.0Hz,1H),6.81(s,1H),6.98(s,1H),7.15-7.25(m,11H),7.35(d,J＝7.6Hz,1H),7.44-7.46(m,6H),8.49(s,1H),9.17(s,1H).MS-ESI[M+H] +＝532。
步骤11-4
Figure PCTCN2018110869-appb-000085
化合物104(80mg)溶于5mL甲醇中，滴加HCl/二氧六环溶液至pH<1,搅拌5min后TLC显示反应完全，滴加稀氨水溶液中和反应至pH＝5-6。乙酸乙酯萃取，浓缩后得到黄色固体60mg,直接进行下步反应。
步骤11-5
Figure PCTCN2018110869-appb-000086
化合物105粗品(60mg)溶解到5mL二氯甲烷中，加入2,6-二甲基吡啶后滴加三氟甲磺酸酐(2.0eq.)。反应10min后TLC显示反应完全，加入水淬灭反应。二氯甲烷萃取后干燥，柱层析得到白色固体30mg。 1H NMR(CDCl 3,400M):δ2.69(s,3H),2.73(s,3H),7.16(dd,J＝8.4,2.0HZ,1H),7.21(s,1H),7.27(d,J＝6.8Hz,1H),7.42(d,J＝6.8Hz,1H),7.51(s,1H),7.61(d,J＝8.4Hz,1H),8.68(d,J＝2.0Hz,1H),9.29(d,J＝2.0Hz,1H).
步骤11-6
Figure PCTCN2018110869-appb-000087
化合物106(30mg)，对硼酸苯甲酸甲酯(24mg)，四(三苯基膦)钯(4.4mg),碳酸钾(21mg)加入到10mLDMF中，抽换氩气三次，100℃反应16h。浓缩后乙酸乙酯萃取反应，有机相以无水硫酸钠干燥。浓缩后柱层析得到黄色固体30mg。 1H NMR(CDCl 3,400M):δ2.70(s,3H),2.74(s,3H),3.89(s,3H),7.23-7.27(m,2H),7.40-7.42(m,2H),7.50-7.52(m, 1H),7.63-7.70(m,2H),7.81(s,1H),8.08(d,J＝8.0Hz,1H),8.69(d,J＝2.0Hz,1H),8.69(d,J＝2.0Hz,1H).
步骤11-7
Figure PCTCN2018110869-appb-000088
化合物108(10mg)溶解到5mL四氢呋喃中，加入5mL甲醇以及10％氢氧化钠溶液5mL，60℃反应3h。TLC显示反应完全，降温后滴加醋酸至pH＝5-6。浓缩后加入乙酸乙酯萃取反应。有机相以水洗涤，无水硫酸钠干燥。浓缩后得到黄色固体7mg。 1H NMR(d6-DMSO,400M):δ2.71(s,3H),2.73(s,3H),3.36(br,1H),7.40(d,J＝6.4Hz,1H),7.54(d,J＝6.8Hz,1H),7.71(d,J＝8.4Hz,1H),7.83-7.84(m,2H),7.92-7.93(m,2H),8.03-8.06(m,2H),8.13(s,1H),9.51(s,1H).
实施例12：化合物LW1005012的合成
Figure PCTCN2018110869-appb-000089
步骤12-1
Figure PCTCN2018110869-appb-000090
按照上述制备化合物LW1005011的实施例11步骤11-6，本实施例用原料邻苯硼酸甲酸甲酯代替对苯硼酸甲酸甲酯，纯化后得到化合物111。 1H NMR(CDCl3,400M):δ2.68(s,3H),2.72(s,3H),3.59(s,3H),7.14-7.25(m,4H),7.35-7.40(m,2H),7.48-7.51(m,2H),7.57(d,J＝8.0Hz,1H),7.79(d,J＝8.0Hz,1H),8.66(d,J＝2.0Hz,1H),9.31(d,J＝2.0Hz,1H).
步骤12-2：
Figure PCTCN2018110869-appb-000091
按照上述制备化合物LW1005011的实施例11步骤11-7，本实施例用化合物111进行水解，纯化后得到化合物LW1005012。 1H NMR(d6-DMSO,400M):δ2.70(s,3H),2.72(s,3H),3.37(br,1H),7.28(d,J＝8.0Hz,1H),7.39(d,J＝6.8Hz,1H),7.47-7.54(m,3H),7.61(t.J＝8.0Hz,1H),7.67(s,1H),7.74-7.77(m,2H),7.83(s,1H),8.77(d,J＝2.0Hz,1H),9.50(d,J＝2.0Hz,1H).
实施例13：化合物LW1005013的合成
Figure PCTCN2018110869-appb-000092
步骤13-1
Figure PCTCN2018110869-appb-000093
按照上述制备化合物LW1005011的实施例11步骤11-6，本实施例用原料间苯硼酸甲酸甲酯代替对苯硼酸甲酸甲酯，纯化后得到化合物112。 1H NMR(CDCl3,400M):δ2.69(s,3H),2.73(s,3H),3.91(s,3H),7.19-7.26(m,2H),7.40(d,J＝7.2Hz,1H),7.47-7.51(m,2H),7.64(d,J＝8.4Hz,1H),7.80-7.82(m,2H),7.98(d,J＝8.0Hz,1H),8.31(s,1H),8.68(d,J＝2.0Hz,1H),9.33(d,J＝2.4Hz,1H).
步骤13-2：
Figure PCTCN2018110869-appb-000094
按照上述制备化合物LW1005011的实施例11步骤11-7，本实施例用化合物112进行水解，纯化后得到化合物LW1005013。 1H NMR(d6-DMSO,400M):δ2.71(s,3H),2.73(s,3H),3.46(br,1H),7.40(d,J＝7.6Hz,1H),7.48(t,J＝8.0Hz,1H),7.53(d,J＝7.2Hz,1H),7.65(d,J＝8.4Hz,1H),7.80-7.83(m,3H),7.90(d,J＝7.2Hz,1H),8.02(s,1H),8.28(s,1H).
实施例14：化合物LW1005014的合成
Figure PCTCN2018110869-appb-000095
步骤14-1
Figure PCTCN2018110869-appb-000096
化合物121(5g)溶解于二氯甲烷中，冰水浴下加入间氯过氧苯甲酸6.16g，温度回复室温反应1h，加入饱和碳酸氢钠溶液淬灭反应。二氯甲烷萃取后，有机相以饱和食盐水洗涤，无水硫酸钠干燥。浓缩后直接进行下步反应。将中间体粗品溶于二氯甲烷中，氩气保护下加入无水DMF(11.6g)后，滴加三氯氧磷4.8g。室温反应过夜。饱和碳酸氢钠淬灭反应。二氯甲烷萃取后，有机相以饱和食盐水洗涤，无水硫酸钠干燥，柱层析得到黄色油状物1.2g。 1H NMR(CDCl 3,400M):δ2.55(s,3H),2.64(s,3H),7.17(d,J＝6.8Hz,1H),7.30(dd,J＝8.8,0.8Hz,1H),7.37(d,J＝7.6Hz,1H),8.14(dd,J＝8.4,0.8Hz,1H).
步骤14-2：
Figure PCTCN2018110869-appb-000097
化合物9-1(1.1g),KI(1.43g)以及乙酰氯(0.91g)加入到10mL乙腈中，氩气保护下加热回流过夜。降温后浓缩，柱层析得到黄色油状物1g。
步骤14-3
Figure PCTCN2018110869-appb-000098
化合物124(368mg),化合物6-5(1.3mmol),Pd(PPh 3) 2Cl 2(91mg),Cs 2CO 3(848mg)加入到10mL二氧六环中，抽换氩气三次后加热到40℃反应16h。TLC显示反应完全，回复室温。将溶剂浓缩后，加入水以及乙酸乙酯萃取，有机相浓缩。浓缩液加入10mL甲醇进行溶解，滴入氯化氢/二氧六环溶液，10min后，调节pH＝5-6。将溶剂浓缩，加入水以及乙酸乙酯萃取，有机相浓缩后柱层析，得到黄色固体210mg。 1H NMR(CDCl 3,400M):δ2.59(s,3H),2.77(s,3H),6.76(dd,J＝8.0,2.0Hz,1H),7.02(d,J＝1.6Hz,1H),7.16(d,J＝7.2Hz,1H),7.39(d,J＝7.2Hz,1H),7.44(d,J＝8.4Hz,1H),7.52(s,1H),7.93(d,J＝8.4Hz,1H),8.29(d,J＝8.8Hz,1H).
步骤14-4：
Figure PCTCN2018110869-appb-000099
化合物126溶解二氯甲烷中，加入2,6-二甲基吡啶。氩气保护下滴加三氟甲磺酸酐(1.5equiv)，0.5h后，TLC显示反应完全。加入水淬灭反应，二氯甲烷萃取，浓缩后柱层析得到黄色固体化合物127。
步骤14-5：
Figure PCTCN2018110869-appb-000100
化合物127(105mg)，双频哪醇硼酸酯(191mg)，二氯二(三苯基膦)钯(17.5mg),碳酸铯(326mg)加入到10mL二氧六环中，抽换氩气三次，100℃反应16h。浓缩后乙酸乙酯萃取反应，有机相以无水硫酸钠干燥。浓缩后柱层析得到黄色固体65mg。 1H NMR(CDCl 3,400M):δ1.31(s,12H),2.59(s,3H),2.76(s,3H),7.18(d,J＝8.4Hz,1H),7.39(d,J＝7.2Hz,1H),7.60-7.64(m,3H),7.99-8.02(m,3H),8.32(d,J＝8.8Hz,1H).MS-ESI[M+H] +＝400.
步骤14-6
Figure PCTCN2018110869-appb-000101
化合物129(30mg)，对碘苯甲酸甲酯(24mg)，四(三苯基膦)钯(4.4mg),碳酸钾(21mg)加入到10mL DMF中，抽换氩气三次，100℃反应16h。浓缩后乙酸乙酯萃取反应，有机相以无水硫酸钠干燥。浓缩后柱层析得到黄色固体30mg。
1H NMR(CDCl 3,400M):δ2.60(s,3H),2.79(s,3H),3.89(s,3H),7.18-7.19(m,1H),7.40(d,J＝7.2Hz,1H),7.50(d,J＝7.6Hz,1H),7.63(s,1H),7.68(d,J＝8.4Hz,3H),7.79(s,1H),8.01(d,J＝8.8Hz,1H),8.06-8.08(m,2H),8.33(d,J＝8.4Hz,1H).MS-ESI[M+H] +＝408.
实施例15：化合物LW1005015的合成
Figure PCTCN2018110869-appb-000102
化合物LW1005014(10mg)溶解到5mL四氢呋喃中，加入5mL甲醇以及10％氢氧化钠溶液5mL，60℃反应3h。TLC显示反应完全，降温后滴加醋酸至pH＝5-6。浓缩后加入乙酸乙酯萃取反应。有机相以水洗涤，无水硫酸钠干燥。浓缩后得到黄色固体7mg。 1H NMR(d6-DMSO,400M):δ2.63(s,3H),2.77(s,3H),3.38(br,1H),7.34(d,J＝7.2Hz,1H),7.54(d,J＝6.8Hz,1H),7.72(dd,J＝8.0,1.2Hz,1H),7.85-7.88(m,2H),7.92-7.94(m,2H),8.05(d,J＝8.0Hz,2H),8.12-8.15(m,2H),8.55(d,J＝8.4Hz,1H).MS-ESI[M+H] +＝ 394.
实施例16：化合物LW1005016的合成
Figure PCTCN2018110869-appb-000103
按照上述制备化合物LW1005014的实施例14的步骤14-6，本实施例用化合物129与间碘酸甲酸甲酯进行偶联，纯化后得到化合物LW1005016。 1H NMR(CDCl 3,400M):δ2.60(s,3H),2.79(s,3H),3.90(s,3H),7.17-7.19(m,2H),7.40(d,J＝7.6Hz,1H),7.49-7.51(m,2H),7.63(s,1H),7.67-7.69(d,J＝8.4Hz,1H),7.79-7.82(m,2H),7.96-8.02(m,1H),8.31-8.34(m,2H).MS-ESI[M+H] +＝408.
实施例17：化合物LW1005017的合成
Figure PCTCN2018110869-appb-000104
按照上述制备化合物LW1005015的实施例15，本实施例用化合物LW1005016进行水解，纯化后得到化合物LW1005017。 1H NMR(d6-DMSO,400M):δ2.64(s,3H),2.78(s,3H),7.34(d,J＝7.2Hz,1H),7.54-7.61(m,2H),7.68(dd,J＝8.0,1.2Hz,1H),7.86-7.88(m,2H),7.94-7.99(m,2H),8.07(s,1H),8.14-8.16(d,J＝8.8Hz,1H),8.30(s,3H),8.57(d,J＝8.8Hz,1H).MS-ESI[M+H] +＝394.
实施例18：化合物LW1005018的合成
Figure PCTCN2018110869-appb-000105
按照上述制备化合物LW1005014的实施例14的步骤14-6，本实施例用化合物129与邻碘酸甲酸甲酯进行偶联，纯化后得到化合物LW1005018。 1H NMR(CDCl 3,400M):δ2.60(s,3H),2.79(s,3H),3.57(s,3H),7.15-7.17(m,1H),7.38-7.41(m,3H),7.48-7.51(m,1H),7.61-7.63(m,2H),7.79(d,J＝7.2Hz,1H),8.01(d,J＝8.8Hz,1H),8.32(d,J＝8.8Hz,1H).MS-ESI[M+H] +＝408.
实施例19：化合物LW1005019的合成
Figure PCTCN2018110869-appb-000106
按照上述制备化合物LW1005015的实施例15，本实施例用化合物LW1005018进行水解，纯化后得到化合物LW1005019。 1H NMR(d6-DMSO,400M):δ2.64(s,3H),2.78(s,3H),3.38(br,1H)，7.30-7.35(m,3H),7.46-7.50(m,2H),7.54-7.57(m,2H),7.70-7.72(m,2H),7.78(d,J＝8.4Hz,1H),7.84(s,1H),8.15(d,J＝8.8Hz,1H),8.56(d,J＝9.2Hz,1H).MS-ESI[M+H] +＝394.
实施例20：化合物LW1005020的合成
Figure PCTCN2018110869-appb-000107
步骤20-1：
Figure PCTCN2018110869-appb-000108
间溴苯酚(2g)，DMAP(160mg),三乙胺(2.4g)溶解到二氯甲烷中，滴加醋酸酐(1.8g)，1h后TLC显示反应完全。加入水淬灭反应，二氯甲烷萃取。浓缩后产物直接进行下步反应。
剧烈搅拌下，乙酰化的中间体以及三氯化铝(2.3g)加入到50mL单口瓶中氩气保护下加热到160℃反应2h，冷却至室温，加入3M盐酸溶液以及乙酸乙酯萃取。有机相以水，饱和氯化钠溶液洗涤后，无水硫酸钠干燥。浓缩后柱层析得到淡黄色固体1g。 1H NMR(CDCl 3,400M):δ2.54(s,3H),6.97(dd,J＝8.4,1.6Hz,1H),7.11(d,J＝1.6Hz,1H),7.51(d,J＝8.4Hz,1H),12.28(s,1H).
步骤20-2：
Figure PCTCN2018110869-appb-000109
化合物133(0.3g)以及化合物134(0.36g)溶于10mL乙醇中，加入KOH(0.25g)，40℃反应2h，TLC显示反应完全。浓缩后水以及乙酸乙酯萃取，有机相以水，饱和氯化钠溶液洗涤后，无水硫酸钠干燥。浓缩后柱层析得到黄色固体0.4g。 1H NMR(CDCl 3,400M):δ2.38(s,3H),7.02(dd,J＝8.4,1.6Hz,1H),7.14(m,2H),7.31-7.36(m,3H),7.44(t,J＝8.0Hz,1H),7.54-7.60(m,3H),7.71-7.73(m,1H),7.77(s,1H),7.93(d,J＝15.6Hz,1H),12.90(s,1H).
步骤20-3：
Figure PCTCN2018110869-appb-000110
化合物135(0.2g)溶于10mL DMSO中，加入I 2(130mg)，加热到160℃反应4h，TLC显示反应完全。加入水搅拌后乙酸乙酯萃取。有机相以水，饱和氯化钠溶液洗涤后，无水硫酸钠干燥。浓缩后柱层析得到白色固体154mg。 1H NMR(CDCl 3,400M):δ2.40(s,3H),6.83(s,1H),7.16-7.19(m,1H),7.33-7.38(m,3H),7.49(dd,J＝8.4,1.6Hz,1H),7.54(d,J＝8.0Hz,1H),7.69-7.71(m,1H),7.75(d,J＝2.0Hz,1H),7.80(d,J＝8.0Hz,1H),8.02-8.05(m,2H).
步骤20-4：
Figure PCTCN2018110869-appb-000111
氩气保护下，化合物136(0.14g)溶于无水四氢呋喃(5mL)。0℃下缓慢滴加四氢铝锂溶液(1.1mL，1.0M，THF)。0.5h后TLC显示反应完全。缓慢滴加乙酸乙酯至无气泡冒出，加入水洗涤，有机相以无水硫酸钠干燥。浓缩后快速柱层析得到白色固体58mg。 1H NMR(d6-DMSO,400M):δ2.40(s,3H),5.54(d,J＝4.0Hz,2H)，5.94(t,J＝4.0Hz,1H)，7.15(d,J＝8.4Hz,1H),7.21(d,J＝8.0Hz,1H),7.25(dd,J＝8.4,2.0Hz,1H)，7.37(t,J＝7.6Hz,1H),7.41(d,J＝2.0Hz,1H),7.51(t,J＝8.0Hz,2H),7.55(s,1H),7.65(d,J＝8.0Hz,1H),7.70(d,J＝8.0Hz,1H),7.85(s,1H).
步骤20-5：
Figure PCTCN2018110869-appb-000112
化合物137(30mg)，邻硼酸苯甲酸甲酯(24mg)，四(三苯基膦)钯(4.4mg),碳酸钾(21mg)加入到10mL DMF中，抽换氩气三次，100℃反应16h。浓缩后乙酸乙酯萃取反应，有机相以无水硫酸钠干燥。浓缩后柱层析得到黄色固体30mg。 1H NMR(d6-DMSO,400M):δ2.39(s,3H),3.62-3.64(m,5H),5.93(t,J＝4.0Hz,1H),6.94(d,J＝7.6Hz,1H),7.07(s,1H),7.21(d,J＝7.2Hz,1H),7.37(t,J＝8.0Hz,1H),7.47-7.55(m,5H),7.61-7.65(m,2H),7.71-7.76(m,2H),7.96(s,1H).
步骤20-6：
Figure PCTCN2018110869-appb-000113
化合物138(90mg)溶于10mL乙酸乙酯中，加入Pd/C(10mg),抽换氩气三次。氢气下室温反应24h，TLC显示反应完全。过滤后浓缩，柱层析得到无色油状物55mg。 1H NMR(CDCl 3,400M):δ1.98-2.25(m,2H),2.35(s,3H),2.77-2.83(m,1H),2.92-3.04(m,1H),3.64(s,3H),5.09(dd,J＝10.0,2.4Hz,1H),6.76(dd,J＝8.0,2.0Hz,1H),6.87(d,J＝1.6Hz,1H),7.04(d,J＝7.6Hz,1H),7.10(d,J＝7.2Hz,1H),7.24-7.48(m,8H),7.59(s,1H),7.72(d,J＝8.4Hz,1H).MS-ESI[M+Na] +＝457.
步骤20-7：
Figure PCTCN2018110869-appb-000114
化合物139(50mg)溶于MeOH/THF(5mL/2mL),加入氢氧化钠溶液(3mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体29mg。 1H NMR(CDCl 3,400M):δ2.42(s,3H),4.13(dd,J＝11.6,9.2Hz,1H),4.43(dd,J＝11.2,2.4Hz,1H)，5.23(dd,J＝5.2,2.4Hz,1H),6.87(dd,J＝8.0,2.0Hz,1H),6.96(d,J＝8.4Hz,1H),7.04(d,J＝1.6Hz,1H),7.17(d,J＝6.8Hz,1H),7.33(t,J＝7.6Hz,1H),7.37-7.41(m,5H),7.49(t,J＝7.6Hz,1H),7.52-7.55(m,1H)，7.58-7.60(m,1H),7.64(s,1H),7.92(d,J＝8.0Hz,1H).
实施例21：化合物LW1005021的合成
Figure PCTCN2018110869-appb-000115
步骤21-1：
Figure PCTCN2018110869-appb-000116
间溴水杨醛溶于10mL四氢呋喃中，冰水浴下加入三乙胺2g，以及MOMCl(0.76mL)。1h后TLC显示反应完全，加入水淬灭反应。乙酸乙酯萃取，有机相以水，饱和氯化钠溶液洗涤后，无水硫酸钠干燥。浓缩后的油状物直接进行下一步反应。
中间体粗品溶解到二氯甲烷20mL中，加入间氯过氧苯甲酸1.73g。室温反应过夜，加入10％氢氧化钠(20mL)搅拌0.5h，醋酸调节pH至5-6。二氯甲烷萃取，干燥浓缩后柱层析得到油状物630mg。 1H NMR(CDCl 3,400M):δ3.44(s,3H),5.10(s,2H),5.93(br,1H),6.87-6.88(m,2H),7.02(d,J＝2.0Hz,1H).
步骤21-2：
Figure PCTCN2018110869-appb-000117
化合物143(339mg)以及化合物144(300mg)溶解到10mL丙酮中。加入碳酸钾330mg，回流3h。TLC显示反应完全。将丙酮浓缩后加入水以及乙酸乙酯萃取。有机相以水，饱和氯化钠溶液洗涤后，无水硫酸钠干燥。浓缩后柱层析,得到黄色油状物420mg。 1H NMR(CDCl 3,400M):δ2.37(s,3H),3.41(s,3H),5.13(s,2H),5.32(s,2H),6.91(s,J＝0.8Hz,1H),6.98-6.99(m,2H),7.15(d,J＝6.8Hz,1H),7.29(t,J＝7.2Hz,1H),7.35(d,J＝8.8Hz,2H),7.50(d,J＝8.0Hz,1H),7.77(d,J＝8.0Hz,1H),7.88(d,J＝8.0Hz,1H),8.12(t,J＝2.0Hz,1H).
步骤21-3：
Figure PCTCN2018110869-appb-000118
化合物145(300mg)溶于10mL甲醇中，加入硼氢化钠38mg，20min后TLC显示反应完全。冰浴下缓慢滴加氯化氢/二氧六环溶液(2mL),滴加完毕后继续搅拌15min，TLC显示反应完全。加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到泡沫状固体222mg。 1H NMR(CDCl 3,400M):δ2.35(s,3H),4.04-4.11(m,4H),5.12-5.13(m,1H),6.76-6.79(m,1H),6.92-6.97(m,2H),7.12(s,1H),7.19(s,1H),7.26-7.39(m,4H),7.50(s,1H),7.56(s,1H)。
步骤21-4：
Figure PCTCN2018110869-appb-000119
化合物146(100mg)溶于10mL四氢呋喃中。氩气保护下加入三苯基膦(102mg)。冰水浴下滴加DIAD(79mg)。保持该温度下反应0.5h，TLC显示反应完全。加入水淬灭反应，乙酸乙酯萃取。有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到无色油状物40mg。 1H NMR(CDCl 3,400M):δ2.36(s,3H),3.99(dd,J＝11.6,9.2Hz,1H),4.33(dd,J＝11.6,2.4Hz,1H),5.08(dd,J＝8.8,2.0Hz,1H),6.81(d,J＝8.4,2.1Hz,1H),6.92(dd,J＝8.8,2.0Hz,1H),7.12(d,J＝7.6Hz,1H),7.25-7.27(m,2H),7.29-7.33(m,3H),7.41(t,J＝8.0Hz,1H),7.54(s,1H).
步骤21-5：
Figure PCTCN2018110869-appb-000120
化合物147(35mg),碳酸钾25mg，2-苯甲酸甲酯硼酸(25mg),Pd(PPh 3) 4(11mg)加入到5mLDMF中，氩气保护下100℃反应16h。减压浓缩后，加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体17mg。 1H NMR(CDCl 3,400M):δ2.37(s,3H),2.64(s,3H),4.03-4.08(m,1H),4.36(d,J＝11.6Hz,1H),5.17(d,J＝8.8Hz,1H),6.77(d,J＝8.4Hz,1H),6.88(s,1H),6.95(d,J＝8.0Hz,1H),7.12(d,J＝6.4Hz,1H),7.26-7.35(m,6H),7.41-7.46(m,2H),7.54(d,J＝7.6Hz,1H),7.58(s,1H),7.71(d,J＝8.0Hz,1H).
步骤21-6：
Figure PCTCN2018110869-appb-000121
化合物148(12mg)溶于MeOH/THF(5mL/2mL),加入氢氧化钠溶液(3mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH＝5-6。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体10mg。 1H NMR(CDCl 3,400M):δ2.35(s,3H),4.01-4.07(m,1H),4.32(dd,J＝11.6,2.4Hz,1H),5.15(dd,J＝9.2,2.0Hz,1H),6.81(dd,J＝7.6,2.0Hz,1H),6.90(d,J＝2.0Hz,1H),6.96(d,J＝8.4Hz,1H),7.11(d,J＝7.2Hz,1H)，7.25-7.35(m,6H),7.40(t,J＝12.0Hz,1H),7.45-7.53(m,2H),7.57(s,1H),7.86(d,J＝7.6Hz,1H).MS-ESI[M+Na] +＝457.
实施例22：化合物LW1005022的合成
Figure PCTCN2018110869-appb-000122
步骤22-1：
Figure PCTCN2018110869-appb-000123
化合物151溶于DMF中，依次加入碳酸钾以及苄基溴，加热至120℃反应2h反应完全，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到无色油状物。 1H NMR(CDCl 3,400M):δ3.48(s,3H),5.09(s,2H),5.17(s,2H),7.00(d,J＝0.8Hz,1H),7.05(s,1H),7.31-7.43(m,5H).
步骤22-2：
Figure PCTCN2018110869-appb-000124
化合物152溶于四氢呋喃中，滴加HCl/二氧六环溶液，室温反应2h后浓缩柱层析得到黄色固体。 1H NMR(CDCl 3,400M):δ5.06(s,2H),6.81(d,J＝8.0Hz,1H),6.99(d,J＝ 8.4,2.0Hz,1H),7.05(d,J＝2.0Hz,1H),7.39-7.41(m,5H).
步骤22-3：
Figure PCTCN2018110869-appb-000125
按照上述制备实施例21的步骤21-2，本实施例用化合物153与化合物154进行反应，纯化后得到化合物155。 1H NMR(CDCl 3,400M):δ2.41(s,3H),5.09(s,2H),5.33(s,2H),6.78(d,J＝8.4Hz,1H),6.98(dd,J＝8.8,2.4Hz,1H),7.06(d,J＝2.0Hz,1H),7.19(d,J＝7.6Hz,1H),7.27-7.39(m,8H),7.49(t,J＝7.6Hz,1H),7.78-7.81(m,1H),7.90-7.92(m,1H),8.14(t,J＝2.0Hz,1H).
步骤22-4：
Figure PCTCN2018110869-appb-000126
按照上述制备实施例21的步骤21-5，本实施例用化合物156与邻苯硼酸甲酸甲酯进行反应，纯化后得到化合物156。 1H NMR(CDCl 3,400M):δ2.41(s,3H),3.55(s,3H),5.12(s,2H),5.40(s,2H)，6.82(dd,J＝6.4,2.0Hz,1H),6.92-6.94(m,2H),7.19(d,J＝6.4Hz,1H),7.25-7.40(m,10H),7.45-7.52(m,2H),7.73(dd,J＝8.0,1.2Hz,1H),7.78-7.81(m,1H),7.96-7.98(m,1H),8.19(t,J＝1.6Hz,1H).
步骤22-5：
Figure PCTCN2018110869-appb-000127
化合物12-4溶解到甲醇中，加入Pd/C(5％)，抽换氢气三次后室温反应过夜，过滤后浓缩得到目标化合物12-5。
步骤22-6：
Figure PCTCN2018110869-appb-000128
按照上述制备实施例21的步骤21-3，本实施例用化合物157进行反应，纯化后得到化合物158。 1H NMR(CDCl 3,400M):δ2.35(s,3H),3.60(s,3H),4.11(d,J＝9.2Hz,1H),4.19(dd,J＝10.0,2.8Hz,1H),5.16(dd,J＝8.6,3.2Hz,1H),6.59(br,1H),6.68(dd,J＝8.0,2.0Hz,1H),6.83(d,J＝8.0Hz,1H),6.88(d,J＝2.0Hz,1H),7.11(d,J＝7.2Hz,1H),7.24-7.44(m,8H),7.50(d,J＝7.6Hz,1H),7.59(s,1H),7.69(d,J＝7.6Hz,1H).
步骤22-7：
Figure PCTCN2018110869-appb-000129
按照上述制备实施例21的步骤21-4，本实施例用化合物158进行反应，纯化后得到化合物159。 1H NMR(CDCl 3,400M):δ2.36(s,3H),3.64(s,3H),4.07(d,J＝9.2Hz,1H),4.36(dd,J＝7.2,2.0Hz,1H),5.16(dd,J＝8.4,1.6Hz,1H),6.68(dd,J＝8.0,2.0Hz,1H),6.76(dd,J＝8.4,2.0Hz,1H),6.89(d,J＝8.0Hz,1H),6.95(d,J＝1.6Hz,1H),7.11(d,J＝7.6Hz,1H),7.25-7.35(m,6H),7.40-7.45(m,2H),7.53(d,J＝7.6Hz,1H),7.58(s,1H),7.70-7.72(m,1H).
步骤22-8：
Figure PCTCN2018110869-appb-000130
按照上述制备实施例21的步骤21-6，本实施例用化合物159进行水解反应，纯化后得到化合物LW1005022。 1H NMR(CDCl 3,400M):δ2.42(s,3H),4.13(dd,J＝11.6,9.2Hz,1H),4.43(dd,J＝11.2,2.4Hz,1H),5.23(dd,J＝9.2,2.4Hz,1H),6.87(dd,J＝8.0,2.0Hz,1H),6.96(d,J＝8.4Hz,1H),7.04(d,J＝1.6Hz,1H),7.17(d,J＝6.8Hz,1H),7.33(t,J＝7.6Hz,1H),7.37-7.41(m,5H),7.47(t,J＝8.0Hz,1H),7.54(td,J＝7.6,1.2Hz,1H),7.58-7.60(m,1H),7.64(s,1H),7.92(d,J＝8.0Hz,1H).
实施例23：化合物LW1005023的合成
Figure PCTCN2018110869-appb-000131
步骤23-1：
Figure PCTCN2018110869-appb-000132
5-溴-2-羟基苯甲醇(0.4g),化合物161(0.49g),以及碳酸钾(552mg)依次加入到20mL丙酮中，加热回流20h(可以缩短时间)。降温后减压浓缩，随后加入水以及乙酸乙酯萃取，有机相以饱和食盐水洗涤，无水硫酸钠干燥。浓缩后柱层析得到化合物350mg。 1H NMR(CDCl 3,400M):δ2.44(s,3H),3.50(br,1H),4.74(s,2H),5.43(s,2H),6.74(d,J＝11.6Hz,1H),7.22-7.26(m,2H),7.33-7.45(m,5H),7.58(t,J＝10.0Hz,1H),7.84-7.92(m,2H),8.14(s,1H).
步骤23-2：
Figure PCTCN2018110869-appb-000133
化合物163(0.3g),三苯基膦氢溴酸盐(254mg,1.0eq.)加入到10mL乙腈中，氩气保护下加热回流24h，TLC显示底物有部分剩余，生成极性很大的产物。浓缩后直接进行下一步。氩气保护下，将少量的钠块加入到10mL无水乙醇中，反应完毕后加入到上步反应的乙醇溶液中，室温反应2h后，TLC显示有明显产物生成。加入水淬灭反应。将乙醇浓缩，随后加入水以及乙酸乙酯萃取，有机相以饱和食盐水洗涤，无水硫酸钠干燥。浓缩后柱层析得到油状化合物140mg。 1H NMR(CDCl 3,400M):δ2.44(s,3H),5.21(s,2H),6.73-6.78(m,2H),7.20-7.22(m,3H),7.34-7.41(m,4H),7.46(t,J＝7.6Hz,1H),7.54-7.56(m,1H),7.60(d,J＝1.6Hz,1H).
步骤23-3：
Figure PCTCN2018110869-appb-000134
化合物165(140mg),邻硼酸苯甲酸甲酯(100mg),Pd(PPh 3) 4(21mg),碳酸钾(102mg)加入到10mLDMF中，抽换氩气三次后加热到100℃反应16h。TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到白色油状物80mg。 1H NMR(CDCl 3,400M):δ2.45(s,3H),3.70(s,3H)，5.27(s,2H),6.89(s,1H),7.08(s,1H),7.20(d,J＝7.6Hz,1H),7.25-7.26(m,1H),7.34-7.55(m,10H),7.63(d,J＝1.2Hz,1H),7.80(dd,J＝6.8,1.6Hz,1H).
步骤23-4:
Figure PCTCN2018110869-appb-000135
化合物166(20mg)溶于MeOH/THF(3mL/2mL),加入氢氧化钠溶液(2mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体15mg。 1H NMR(CDCl 3,400M):δ2.42(s,3H),3.06-3.21(m,2H),3.34-3.39(m,1H),3.14(t,J＝10.8Hz,1H),4.44 (d,J＝10.4Hz,1H),6.90(d,J＝8.0Hz,1H),7.12-7.14(m,2H),7.18(d,J＝6.8Hz,1H),7.23(d,J＝7.2Hz,1H),7.31-7.56(m,9H),7.92(d,J＝7.6Hz,1H).MS-ESI:[M-H] +＝419.
步骤23-5：
Figure PCTCN2018110869-appb-000136
化合物167(20mg)溶于MeOH/THF(3mL/2mL),加入氢氧化钠溶液(2mL,10％)，60℃反应5h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体15mg。 1H NMR(CDCl 3,400M):δ2.42(s,3H),3.06-3.21(m,2H),3.34-3.39(m,1H),3.14(t,J＝10.8Hz,1H),4.44(d,J＝10.4Hz,1H),6.90(d,J＝8.0Hz,1H),7.12-7.14(m,2H),7.18(d,J＝6.8Hz,1H),7.23(d,J＝7.2Hz,1H),7.31-7.56(m,9H),7.92(d,J＝7.6Hz,1H).MS-ESI:[M-H] +＝419.
实施例24：化合物LW1005024合成
Figure PCTCN2018110869-appb-000137
步骤24-1：
Figure PCTCN2018110869-appb-000138
50mL单口瓶中依次加入化合物171(0.4g)，Pd(PPh 3) 2Cl 2(15mg)，CuI(4mg)，THF(5mL)以及三乙胺(2.5mL)抽换氩气后加入5-溴-2-碘苯甲酸甲酯(710mg)。室温反应16h，TLC显示反应完全。加入少量水淬灭反应。浓缩后柱层析得到亮黄色油状物316mg。 1H NMR(CDCl 3,400M):δ2.43(s,3H),3.97(s,3H),7.19(d,J＝7.6Hz,1H),7.34(t,J＝7.6Hz,1H),7.40-7.45(m,3H),7.52-7.59(m,3H),7.63(dd,J＝8.8,2.4Hz,1H),7.78(s,1H),8.13(d,J＝2.0Hz,1H).MS-ESI:[M+H] +＝406.
步骤24-2：
Figure PCTCN2018110869-appb-000139
化合物172(300mg)溶于MeOH/THF(10mL/5mL),加入氢氧化钠溶液(5mL,10％)，60℃反应1h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体230mg。 1H NMR(CDCl 3,400M):δ2.42(s,3H),7.18(d,J＝7.2Hz,1H),7.31-7.41(m,4H),7.52(d,J＝8.0Hz,1H),7.56(d,J＝8.0Hz,1H),7.69(dd,J＝8.4,2.0Hz,1H),7.79(s,1H),8.25(d,J＝1.6Hz,1H).
步骤24-3：
Figure PCTCN2018110869-appb-000140
化合物173(100mg)加入到10mL甲苯中加入2滴三氟甲磺酸，加热到110℃反应2h，TLC显示反应完全，降温后加入水淬灭反应，乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到淡黄色固体83mg。 1H NMR(CDCl 3,400M):δ2.46(s,3H),7.00(s,1H),7.22(d,J＝7.2Hz,1H),7.37(d,J＝7.6Hz,1H),7.41(d,J＝8.8Hz,1H),7.44(d,J＝8.8Hz,1H),7.53(t,J＝7.6Hz,1H),7.65-7.68(m,1H),7.82-7.85(m,2H),8.07(t,J＝1.6Hz,1H),8.47(d,J＝2.0Hz,1H).
步骤24-4：
Figure PCTCN2018110869-appb-000141
干燥的50mL三口瓶中加入化合物174(80mg)加入到无水THF中，0℃下滴加LiAlH 4(0.4mL，1M in THF)。维持该温度反应2h，加入乙酸乙酯淬灭反应，随后加入水以及乙酸乙酯，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥，柱层析得到固体43mg。 1H NMR(CDCl 3,400M):δ2.46(s,3H),3.04(dd,J＝14.4,4.0Hz,1H),3.12(dd,J＝14.4,8.8Hz,1H),4.48(d,J＝12.4Hz,1H),4.74(d,J＝12.0Hz,1H),4.96(dd,J＝8.8,4.0Hz,1H)7.07(d,J＝8.0Hz,1H),7.32-7.46(m,6H),7.52-7.54(m,3H).
步骤24-5：
Figure PCTCN2018110869-appb-000142
化合物175(260mg)，三苯基膦(516mg)溶解到10mL无水THF中。降温至0℃，滴加偶氮二甲酸二异丙酯(398mg)。滴加完毕后，维持该温度反应1h，TLC显示反应完全，加入水淬灭反应。乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥，柱层析得到无色油状物65mg。 1H NMR(CDCl 3,400M):δ2.41(s,3H),2.94-3.08(m,2H),4.74(dd,J＝10.4,3.6Hz,1H),4.97(d,J＝2.8Hz,2H),7.02(d,J＝8.0Hz,1H),7.16(d,J＝7.6Hz,1H),7.22(s,1H),7.32(t,J＝8.0Hz,1H),7.37-7.46(m,4H),7.52-7.54(m,1H),7.64(s,1H).
步骤24-6：
Figure PCTCN2018110869-appb-000143
化合物176(60mg),邻硼酸苯甲酸甲酯(43mg),Pd(PPh 3) 4(9mg),碳酸钾(44mg)加入到5mLDMF中，抽换氩气三次后加热到100℃反应16h。TLC显示反应完全，降温后，减压浓缩DMF，加入水以及乙酸乙酯萃取，无水硫酸钠干燥。柱层析得到无色油状物35mg。 1H NMR(CDCl 3,400M):δ2.43(s,3H),3.05-3.21(m,2H),3.70(s,3H),4.84(dd,J＝7.6,2.8Hz,1H),5.07(s,2H),7.05(s,1H),7.16-7.19(m,3H),7.22(s,1H),7.34-7.56(m,9H),7.69(s,1H),7.83(d,J＝6.8Hz,1H).
步骤24-7：
Figure PCTCN2018110869-appb-000144
化合物178(35mg)溶于MeOH/THF(3mL/2mL),加入氢氧化钠溶液(4mL,10％)，60℃反应3h。降温后滴加10％硫酸溶液至pH<7。浓缩后加入水以及乙酸乙酯萃取，有机相以水以及饱和食盐水洗涤，无水硫酸钠干燥。柱层析得到白色固体28mg。 1H NMR(CDCl 3,400M):δ2.42(s,3H),3.03-3.20(m,2H),3.70(s,3H),4.83(dd,J＝7.6,3.6Hz,1H),5.06(s,2H),7.07(s,1H),7.15-7.20(m,3H),7.22(s,1H),7.31-7.46(m,7H),7.53-7.58(m,2H),7.68(s,1H),7.96(d,J＝8.0Hz,1H).MS-ESI:[M-H] +＝419.
测试例1:检测化合物对PD-1/PD-L1蛋白相互结合的抑制效应
通过均质时间分辨荧光分析(HTRF)法，检测化合物结合PD-L1以及抑制PD-1/PD-L1相互结合的能力。
试剂和耗材
PD1/PDL1结合检测试剂盒(PD1/PDL1 binding assay kit-indirect format 1000tests，Cat.No.:64CUS000C-1，Cisbio)
Figure PCTCN2018110869-appb-000145
Figure PCTCN2018110869-appb-000146
溶液配制
Figure PCTCN2018110869-appb-000147
实验方法
化合物按梯度稀释，在4μL Tag1-PD-L1蛋白(终浓度为2nM)中加入2μL的化合物或者稀释缓冲液，混合液在常温环境下预孵育15分钟。接着加入4μL Tag2-PD1蛋白(终浓度为10nM)，进一步孵育15分钟。随后加入5μL抗Tag1-Eu 3+抗体和5μL抗Tag2-XL665抗体或者10μL抗Tag1-Eu 3+抗体和抗Tag2-XL665抗体的预混合物。封板，反应液常温反应2小时，EnVision读板器上检测665nm和620nm荧光信号。
通过665nm和620nm的荧光发射值计算HTRF比率。
HTRF比率＝(665nm/620nm)x 10 4
用下列公式来计算检测化合物的抑制率(Inhibition rate，IR)：IR(％)＝((Ratio溶剂对照–Ratio化合物)/(Ratio溶剂对照–Ratio空白对照))*100％。
表2:本发明的化合物抑制PD-1的活性测试结果
NO. 化合物 抑制率％(@8nM)
1 LW1005006 14.6
2 LW1005009 21.7
3 LW1005011 23.0
4 LW1005012 22.7
5 LW1005013 27.4
6 LW1005014 22.8
7 LW1005015 15.3
8 LW1005016 19.4
9 LW1005017 20.1
10 LW1005018 19.4
11 LW1005019 24.2
12 LW1005020 29.5
13 LW1005021 18.0
14 LW1005022 20.6
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (10)

  1. 一种如式L所示的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物：
    Figure PCTCN2018110869-appb-100001
    其中，环A由环B和环C并环形成，如式L-1和式L-2所示：
    Figure PCTCN2018110869-appb-100002
    其中，
    R 1、R 2、R 3、R 4各自独立地为氢、氧代(O或＝O)、卤素、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、取代或未取代的C3-C8环烷基、-CN、羟基、氨基、羧基、取代或未取代的C6-C10芳基、取代或未取代的C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、卤代的具有1-3个选自N、S和O的杂原子的5-10元杂芳基，或-(CH 2) n-Ar；其中，n为0、1、2、3或4；Ar为未取代的或取代的C6-C10芳基、未取代的或取代的5-10元杂芳基、未取代的或取代的5-10元杂芳基；所述取代的是指被选自下组的一个或多个取代基所取代：羧基(-COOH)、酯基(C1-C6烷基O(CO)-)；或为无；
    环B与环C各自独立地为5-6元芳香环、5-6元芳香杂环、5-6元饱和脂肪环或饱或5-6元脂肪杂环；且所述的环B和环C组成并环结构；
    X为无(即R 5和环C或环B直接相连)、5-10元的芳基或杂芳基；
    R 5为未取代的或取代的选自下组的基团：5-10元芳基、5-10元杂芳基、5-10元环基、5-10元杂环基；所述取代的是指被选自下组的一个或多个取代基所取代：C1-C6烷基、卤代C1-C6烷基、羧基(-COOH)、酯基(C1-C6烷基O(CO)-)；
    所述杂环基、杂芳基、芳香杂环或饱和脂肪杂环包含1-3个(1个，2个或3个)选自N、S和O的杂原子；
    除非特别说明，所述的取代指基团上的一个或多个氢原子被选自下组的取代基所取代：卤素、C1-C6烷基、卤代的C1-C6烷基、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、氧代、-CN、羟基、氨基、羧基、C6-C10芳基、卤代的C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、卤代的具有1-3个选自N、S和O的杂原子的5-10元杂芳基。
  2. 如权利要求1所述的化合物、其立体异构体或其互变异构体，或其药学上可接受 的盐、水合物或溶剂化物，其特征在于，
    R 5-X-为选自下组的结构：
    Figure PCTCN2018110869-appb-100003
    其中，R 10、R 11、R 12、R m、R n、R L各自独立地选自下组：H、卤素、C1-C6烷基、卤代的C1-C6烷基、C1-C6烷氧基、卤代的C1-C6烷氧基、C3-C8环烷基、卤代的C3-C8环烷基、氧代、-CN、羟基、氨基、羧基、C6-C10芳基、卤代的C6-C10芳基、具有1-3个选自N、S和O的杂原子的5-10元杂芳基、卤代的具有1-3个选自N、S和O的杂原子的5-10元杂芳基。
  3. 如权利要求1所述的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物，其特征在于，式L所示的化合物选自下组：
    Figure PCTCN2018110869-appb-100004
  4. 如权利要求1所述的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物，其特征在于，式L所示的化合物选自下组：
    Figure PCTCN2018110869-appb-100005
    Figure PCTCN2018110869-appb-100006
    各式中，各基团的定义同前。
  5. 如权利要求1所述的化合物，其特征在于，所述的化合物选自下组：
    Figure PCTCN2018110869-appb-100007
  6. 权利要求1所述化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物的制备方法，其特征在于，
    (1)所述化合物为式I-5化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100008
    或(2)所述化合物为式II-5化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100009
    或(3)所述化合物为式III-5化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100010
    或(4)所述化合物为式IV-7化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100011
    或(5)所述化合物为式V-7化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100012
    或(6)所述化合物为式VI-8化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100013
    或(7)所述化合物为式VII-4化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100014
    或(8)所述化合物为式VIII-8化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100015
    或(9)所述化合物为式IX-7化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100016
    或(10)所述化合物为式X-6化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100017
    或(11)所述化合物为式XI-7化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100018
    或(12)所述化合物为式XII-7化合物，所述方法包括步骤：
    Figure PCTCN2018110869-appb-100019
    各式中，各个基团的定义同前。
  7. 如下各式所示的中间体，其立体异构体、顺反异构体或互变异构体，
    Figure PCTCN2018110869-appb-100020
    Figure PCTCN2018110869-appb-100021
    Figure PCTCN2018110869-appb-100022
    各式中，R 1、R 2、R 3、R 4和R 5的定义同前。
  8. 一种药物组合物，其特征在于，包含(1)如权利要求1所述的化合物、其立体异构体或互变异构体、或其药学上可接受的盐、水合物或溶剂化物；(2)药学上可接受的载体。
  9. 如权利要求1所述的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物或如权利要求7所述药物组合物的用途，其特征在于，用于制备PD-1/PD-L1抑制剂。
  10. 如权利要求1所述的化合物、其立体异构体或其互变异构体，或其药学上可接受的盐、水合物或溶剂化物或如权利要求7所述药物组合物的用途，其特征在于，用于制备治疗癌症的药物；优选地，所述癌症选自下组：黑色素瘤、肺癌、膀胱癌。
PCT/CN2018/110869 2017-10-19 2018-10-18 Pd-1/pd-l1小分子抑制剂及其制备方法和用途 WO2019076343A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201710980409.8A CN109678796B (zh) 2017-10-19 2017-10-19 Pd-1/pd-l1小分子抑制剂及其制备方法和用途
CN201710980409.8 2017-10-19

Publications (1)

Publication Number Publication Date
WO2019076343A1 true WO2019076343A1 (zh) 2019-04-25

Family

ID=66173536

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/110869 WO2019076343A1 (zh) 2017-10-19 2018-10-18 Pd-1/pd-l1小分子抑制剂及其制备方法和用途

Country Status (2)

Country Link
CN (1) CN109678796B (zh)
WO (1) WO2019076343A1 (zh)

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020023710A1 (en) 2018-07-27 2020-01-30 Arbutus Biopharma Corporation Substituted tetrahydrocyclopenta[c]pyrroles, substituted dihydropyrrolizines, analogues thereof, and methods using same
WO2020123674A1 (en) 2018-12-12 2020-06-18 Arbutus Biopharma Corporation Substituted arylmethylureas and heteroarylmethylureas, analogues thereof, and methods using same
US10710986B2 (en) 2018-02-13 2020-07-14 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
US10774071B2 (en) 2018-07-13 2020-09-15 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
US10899735B2 (en) 2018-04-19 2021-01-26 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
US11059834B2 (en) 2017-08-08 2021-07-13 Chemocentryx, Inc. Macrocyclic immunomodulators
US11130740B2 (en) 2017-04-25 2021-09-28 Arbutus Biopharma Corporation Substituted 2,3-dihydro-1H-indene analogs and methods using same
US11135210B2 (en) 2018-02-22 2021-10-05 Chemocentryx, Inc. Indane-amines as PD-L1 antagonists
US11236085B2 (en) 2018-10-24 2022-02-01 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
US11266643B2 (en) 2019-05-15 2022-03-08 Chemocentryx, Inc. Triaryl compounds for treatment of PD-L1 diseases
CN114746417A (zh) * 2019-12-06 2022-07-12 南京明德新药研发有限公司 Pan-RAF激酶抑制剂的联芳基化合物
US11426364B2 (en) 2016-06-27 2022-08-30 Chemocentryx, Inc. Immunomodulator compounds
US11485708B2 (en) 2019-06-20 2022-11-01 Chemocentryx, Inc. Compounds for treatment of PD-L1 diseases
US11708326B2 (en) 2017-07-28 2023-07-25 Chemocentryx, Inc. Immunomodulator compounds
US11713307B2 (en) 2019-10-16 2023-08-01 Chemocentryx, Inc. Heteroaryl-biphenyl amides for the treatment of PD-L1 diseases
US11866429B2 (en) 2019-10-16 2024-01-09 Chemocentryx, Inc. Heteroaryl-biphenyl amines for the treatment of PD-L1 diseases
US11872217B2 (en) 2019-07-10 2024-01-16 Chemocentryx, Inc. Indanes as PD-L1 inhibitors
US12083118B2 (en) 2018-03-29 2024-09-10 Arbutus Biopharma Corporation Substituted 1,1′-biphenyl compounds, analogues thereof, and methods using same

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110272417B (zh) * 2019-06-18 2021-07-13 五邑大学 2-甲基-1,8-萘啶类化合物及其制备方法与应用
CN112457331A (zh) * 2019-09-09 2021-03-09 上海长森药业有限公司 含氮杂环类化合物，及其制备方法、药物组合物和应用
CN112479988B (zh) * 2020-12-09 2024-05-03 药康众拓(江苏)医药科技有限公司 取代联苯类化合物及其制备方法、用途和药物组合物
CN114773309B (zh) * 2022-04-12 2024-06-28 乐威医药（江苏）股份有限公司 1-（7-卤苯并[d][1,3]二氧杂环-4-基）烷基酮类化合物的制备方法及应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104159911A (zh) * 2012-03-07 2014-11-19 奥瑞基尼探索技术有限公司 作为免疫调节剂的模拟肽化合物
WO2015160641A2 (en) * 2014-04-14 2015-10-22 Bristol-Myers Squibb Company Compounds useful as immunomodulators
CN105849092A (zh) * 2013-09-06 2016-08-10 奥瑞基尼探索技术有限公司 作为免疫调节剂的1，3，4-*二唑和1，3，4-噻二唑衍生物
WO2017066227A1 (en) * 2015-10-15 2017-04-20 Bristol-Myers Squibb Company Compounds useful as immunomodulators
WO2017106634A1 (en) * 2015-12-17 2017-06-22 Incyte Corporation N-phenyl-pyridine-2-carboxamide derivatives and their use as pd-1/pd-l1 protein/protein interaction modulators

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104159911A (zh) * 2012-03-07 2014-11-19 奥瑞基尼探索技术有限公司 作为免疫调节剂的模拟肽化合物
CN105849092A (zh) * 2013-09-06 2016-08-10 奥瑞基尼探索技术有限公司 作为免疫调节剂的1，3，4-*二唑和1，3，4-噻二唑衍生物
WO2015160641A2 (en) * 2014-04-14 2015-10-22 Bristol-Myers Squibb Company Compounds useful as immunomodulators
WO2017066227A1 (en) * 2015-10-15 2017-04-20 Bristol-Myers Squibb Company Compounds useful as immunomodulators
WO2017106634A1 (en) * 2015-12-17 2017-06-22 Incyte Corporation N-phenyl-pyridine-2-carboxamide derivatives and their use as pd-1/pd-l1 protein/protein interaction modulators

Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11793771B2 (en) 2016-06-27 2023-10-24 Chemocentryx, Inc. Immunomodulator compounds
US11426364B2 (en) 2016-06-27 2022-08-30 Chemocentryx, Inc. Immunomodulator compounds
US11130740B2 (en) 2017-04-25 2021-09-28 Arbutus Biopharma Corporation Substituted 2,3-dihydro-1H-indene analogs and methods using same
US11708326B2 (en) 2017-07-28 2023-07-25 Chemocentryx, Inc. Immunomodulator compounds
US11059834B2 (en) 2017-08-08 2021-07-13 Chemocentryx, Inc. Macrocyclic immunomodulators
US11691985B2 (en) 2017-08-08 2023-07-04 Chemocentryx, Inc. Macrocyclic immunomodulators
US11555029B2 (en) 2018-02-13 2023-01-17 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
US10710986B2 (en) 2018-02-13 2020-07-14 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
US11759458B2 (en) 2018-02-22 2023-09-19 Chemocentryx, Inc. Indane-amines as PD-L1 antagonists
US11135210B2 (en) 2018-02-22 2021-10-05 Chemocentryx, Inc. Indane-amines as PD-L1 antagonists
US12083118B2 (en) 2018-03-29 2024-09-10 Arbutus Biopharma Corporation Substituted 1,1′-biphenyl compounds, analogues thereof, and methods using same
US10899735B2 (en) 2018-04-19 2021-01-26 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
US10774071B2 (en) 2018-07-13 2020-09-15 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
WO2020023710A1 (en) 2018-07-27 2020-01-30 Arbutus Biopharma Corporation Substituted tetrahydrocyclopenta[c]pyrroles, substituted dihydropyrrolizines, analogues thereof, and methods using same
US11236085B2 (en) 2018-10-24 2022-02-01 Gilead Sciences, Inc. PD-1/PD-L1 inhibitors
WO2020123674A1 (en) 2018-12-12 2020-06-18 Arbutus Biopharma Corporation Substituted arylmethylureas and heteroarylmethylureas, analogues thereof, and methods using same
US11266643B2 (en) 2019-05-15 2022-03-08 Chemocentryx, Inc. Triaryl compounds for treatment of PD-L1 diseases
US11485708B2 (en) 2019-06-20 2022-11-01 Chemocentryx, Inc. Compounds for treatment of PD-L1 diseases
US11872217B2 (en) 2019-07-10 2024-01-16 Chemocentryx, Inc. Indanes as PD-L1 inhibitors
US11713307B2 (en) 2019-10-16 2023-08-01 Chemocentryx, Inc. Heteroaryl-biphenyl amides for the treatment of PD-L1 diseases
US11866429B2 (en) 2019-10-16 2024-01-09 Chemocentryx, Inc. Heteroaryl-biphenyl amines for the treatment of PD-L1 diseases
CN114746417B (zh) * 2019-12-06 2023-12-08 齐鲁制药有限公司 Pan-RAF激酶抑制剂的联芳基化合物
CN114746417A (zh) * 2019-12-06 2022-07-12 南京明德新药研发有限公司 Pan-RAF激酶抑制剂的联芳基化合物

Also Published As

Publication number Publication date
CN109678796A (zh) 2019-04-26
CN109678796B (zh) 2023-01-10

Similar Documents

Publication Publication Date Title
WO2019076343A1 (zh) Pd-1/pd-l1小分子抑制剂及其制备方法和用途
KR102585860B1 (ko) Ｒｈｏ-관련 단백질 키나아제 억제제, 이를 포함하는 약학 조성물, 및 이의 제조 방법 및 용도
AU2017268536B2 (en) Novel compounds and compositions for inhibition of FASN
AU2014262622B2 (en) Benzimidazole derivatives as bromodomain inhibitors
KR102563325B1 (ko) Ｒｈｏ-관련 단백질 키나아제 억제제, Ｒｈｏ-관련 단백질 키나아제 억제제를 함유하는 약학 조성물, 제조 방법 및 약학 조성물의 용도
CA2859604C (en) Compounds
US20200407357A1 (en) Heterocyclic compounds as immunomodulators
WO2021087018A1 (en) Pyridazinones as parp7 inhibitors
AU2020241258A1 (en) Novel small molecule inhibitors of tead transcription factors
SA516371448B1 (ar) مشتقات حلقية غير متجانسة ثنائية الحلقة كمثبطات نطاق برومو
CN110041316B (zh) Tam家族激酶/和csf1r激酶抑制剂及其用途
JP2005521698A (ja) 新規な三環式化合物
JP2016525075A (ja) ヘテロアリール置換ピラゾール類
JP2016536311A (ja) ヘテロアリール置換ピラゾール類
JP2016514718A (ja) ３−ヘテロアリール置換インダゾール類
EP2921480A1 (en) Nitrogen-containing heterocyclic compound
AU2015346223A1 (en) Methods and compositions for inhibition of bromodomain and extraterminal proteins
KR20220062053A (ko) 신규 트리시클릭 방향족 헤테로시클릭 화합물, 및 이의 제조 방법, 약학적 조성물과 응용
CA3191829A1 (en) Cd73 inhibitor and application thereof in medicine
TWI705965B (zh) 新穎三環化合物
EP3383877B1 (en) Heterocycle compounds and uses thereof
JPH03120276A (ja) モノ環式β―ラクタム抗生物質の複素環式ヒドラジド誘導体
EP3749671A1 (en) [1,2,4]triazolo[4,3-a]pyrazin-8-one derivatives
KR20190128671A (ko) 중수소화 벤즈이미다졸 화합물 및 그의 의약 용도
CN111065635B (zh) 作为mth1抑制剂的新型嘧啶衍生物

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18868202

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18868202

Country of ref document: EP

Kind code of ref document: A1