WO2021147965A1 - Macrocyclic compound serving as kras inhibitor - Google Patents

Macrocyclic compound serving as kras inhibitor Download PDF

Info

Publication number
WO2021147965A1
WO2021147965A1 PCT/CN2021/073149 CN2021073149W WO2021147965A1 WO 2021147965 A1 WO2021147965 A1 WO 2021147965A1 CN 2021073149 W CN2021073149 W CN 2021073149W WO 2021147965 A1 WO2021147965 A1 WO 2021147965A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
present
pharmaceutically acceptable
acceptable salt
crude product
Prior art date
Application number
PCT/CN2021/073149
Other languages
French (fr)
Chinese (zh)
Inventor
李秋
王建非
胡国平
付志飞
陈健
孙继奎
张杨
黎健
陈曙辉
Original Assignee
南京明德新药研发有限公司
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 南京明德新药研发有限公司 filed Critical 南京明德新药研发有限公司
Publication of WO2021147965A1 publication Critical patent/WO2021147965A1/en

Links

Images

Classifications

    • 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
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • 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
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the present invention relates to a series of macrocyclic compounds and their application in the preparation of drugs for diseases related to KRAS inhibitors. Specifically, it relates to a compound represented by formula (I) and a pharmaceutically acceptable salt thereof.
  • the first RAS oncogene was found in rat sarcoma (rat sarcoma), hence the name.
  • the RAS protein is a product expressed by the RAS gene, which refers to a closely related monomer globulin consisting of 189 amino acids with a molecular weight of 21KDa.
  • RAS protein can be combined with guanine trinucleotide phosphate (GTP) or guanine dinucleotide phosphate (GDP).
  • GTP guanine trinucleotide phosphate
  • GDP guanine dinucleotide phosphate
  • the active state of RAS protein has an impact on cell growth, differentiation, cytoskeleton, protein transport and secretion, etc.
  • GTP or GDP When the RAS protein binds to GDP, it is in a dormant state, that is, "inactive"; when stimulated by a specific upstream cell growth factor, the RAS protein is induced Exchange GDP and combine with GTP, which is called “activated” state at this time.
  • the RAS protein bound to GTP can activate downstream proteins for signal transmission.
  • RAS protein itself has weak GTP hydrolysis activity and can hydrolyze GTP to GDP. In this way, the transformation from the activated state to the inactivated state can be realized. In this hydrolysis process, GAP (GTPase activating proteins) is also required to participate. It can interact with RAS protein and greatly promote its ability to hydrolyze GTP to GDP.
  • RAS protein will affect its interaction with GAP, which will also affect its ability to hydrolyze GTP to GDP, making it always in an activated state.
  • the activated RAS protein continuously gives growth signals to downstream proteins, which eventually leads to the continuous growth and differentiation of cells, and eventually tumors.
  • RAS gene family There are many members of the RAS gene family, among which the subfamilies closely related to various cancers are mainly Kirsten rat sarcoma virus oncogene homolog (KRAS), Harvey rat sarcoma virus carcinogenic homolog (HRAS) and nerve The blastoma rat sarcoma virus oncogene homolog (NRAS).
  • KRAS Kirsten rat sarcoma virus oncogene homolog
  • HRAS Harvey rat sarcoma virus carcinogenic homolog
  • NRAS blastoma rat sarcoma virus oncogene homolog
  • KRAS G12C mutation is a more common subtype of KRAS gene mutation, which refers to the mutation of glycine 12 to cysteine.
  • KRAS G12C mutation is the most common in lung cancer. According to data reported in the literature (Nat Rev Drug Discov 2014; 13:828-851), KRAS G12C mutation accounts for about 10% of all lung cancer patients.
  • KRAS G12C mutant protein is a cutting-edge target, and there are not many studies at present.
  • the pyridine and the mother core are inserted into the hydrophobic pocket with a dihedral angle of 86.8°, and the fluorophenol and the mother core are inserted into the hydrophobic pocket with a dihedral angle of 58.8°.
  • the phenolic hydroxyl group and Arg68 form a hydrogen bond. (See Figure 1 and Figure 2, Maestro 2017-2, Pymol 1.8.6).
  • the present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
  • R 1 is selected H, F, Cl, Br, I and CH 3, CH 3 optionally substituted by the two or three R a;
  • R 2 is selected from H, F, Cl, Br and I;
  • R 3 is selected from H, F, Cl, Br, and C 1-3 alkyl, said C 1-3 alkyl optionally substituted with 1, 2 or 3 R b;
  • R 4 is selected from H, F, Cl, Br and I;
  • R 5 is selected from C 1-6 alkyl and cyclopropyl, the C 1-6 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c ;
  • n is selected from 1, 2, 3, 4, 5 and 6;
  • n is selected from 0 and 1;
  • p is selected from 1 and 2;
  • R 6 is selected from H and CH 3 ;
  • R a, R b and R c are each independently selected from H, F, Cl, Br and I;
  • R d is selected from H, F, Cl, Br, I and CH 3 ;
  • the carbon atom with "*" is a chiral carbon atom and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.
  • the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
  • R 1 , R 2 , R 3 , R 4 , R 5 and L 1 are as defined in the present invention.
  • R 1 is selected from H, and other variables are as defined in the present invention.
  • R 2 is selected from H, and other variables are as defined in the present invention.
  • R 3 is selected from CH 3 , and other variables are as defined in the present invention.
  • R 4 is selected from F, and other variables are as defined in the present invention.
  • R 5 is selected from C 1-4 alkyl and cyclopropyl, the C 1-4 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c , and others
  • the variables are as defined in the present invention.
  • R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , The CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , Optionally substituted by 1, 2 or 3 R c , and other variables are as defined in the present invention.
  • R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , Other variables are as defined in the present invention.
  • the aforementioned m is selected from 1, 2, 3, 4, and 5, and other variables are as defined in the present invention.
  • the present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
  • R 1 is selected H, F, Cl, Br, I and CH 3, CH 3 optionally substituted by the two or three R a;
  • R 2 is selected from H, F, Cl, Br and I;
  • R 3 is selected from H, F, Cl, Br, and C 1-3 alkyl, said C 1-3 alkyl optionally substituted with 1, 2 or 3 R b;
  • R 4 is selected from H, F, Cl, Br and I;
  • R 5 is selected from C 1-6 alkyl and cyclopropyl, the C 1-6 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c ;
  • n is selected from 1, 2, 3, 4, 5 and 6;
  • n is selected from 0 and 1;
  • p is selected from 1 and 2;
  • R 6 is selected from H and CH 3 ;
  • R a, R b, R c and R d are each independently selected from H, F, Cl, Br and I;
  • the carbon atom with "*" is a chiral carbon atom and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.
  • the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
  • R 1 , R 2 , R 3 , R 4 , R 5 and L 1 are as defined in the present invention.
  • R 1 is selected from H, and other variables are as defined in the present invention.
  • R 2 is selected from H, and other variables are as defined in the present invention.
  • R 3 is selected from CH 3 , and other variables are as defined in the present invention.
  • R 4 is selected from F, and other variables are as defined in the present invention.
  • R 5 is selected from C 1-4 alkyl and cyclopropyl, the C 1-4 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c , and others
  • the variables are as defined in the present invention.
  • R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , The CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , Optionally substituted by 1, 2 or 3 R c , and other variables are as defined in the present invention.
  • R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , Other variables are as defined in the present invention.
  • the aforementioned m is selected from 1, 2, 3, and 4, and other variables are as defined in the present invention.
  • the present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
  • R 1 is selected H, F, Cl, Br, I and CH 3, CH 3 optionally substituted by the two or three R a;
  • R 2 is selected from H, F, Cl, Br and I;
  • R 3 is selected from H, F, Cl, Br, and C 1-3 alkyl, said C 1-3 alkyl optionally substituted with 1, 2 or 3 R b;
  • R 4 is selected from H, F, Cl, Br and I;
  • R 5 is selected from C 1-6 alkyl and cyclopropyl, the C 1-6 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c ;
  • L 1 is selected from -(CH 2 ) m -, -(CH 2 ) m -NH-, -(CH 2 ) m -O-, -NH-(CH 2 ) m -NH-, -NH-(CH 2 ) m -O- and -O-(CH 2 ) m -O-, the -(CH 2 ) m -, -(CH 2 ) m -NH-, -(CH 2 ) m -O-, -NH -(CH 2 ) m -NH-, -NH-(CH 2 ) m -O- and -O-(CH 2 ) m -O- are optionally substituted by 1, 2 or 3 R d ;
  • n is selected from 2, 3, 4, 5 and 6;
  • R a, R b, R c and R d are each independently selected from H, F, Cl, Br and I;
  • the carbon atom with "*" is a chiral carbon atom and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.
  • the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
  • R 1 , R 2 , R 3 , R 4 , R 5 and L 1 are as defined in the present invention.
  • R 1 is selected from H, and other variables are as defined in the present invention.
  • R 2 is selected from H, and other variables are as defined in the present invention.
  • R 3 is selected from CH 3 , and other variables are as defined in the present invention.
  • R 4 is selected from F, and other variables are as defined in the present invention.
  • R 5 is selected from C 1-4 alkyl and cyclopropyl, the C 1-4 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c , and others
  • the variables are as defined in the present invention.
  • R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , The CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , Optionally substituted with 1, 2 or 3 R c .
  • R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 , Other variables are as defined in the present invention.
  • the above m is selected from 2, 3 and 4, and other variables are as defined in the present invention.
  • the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-,- NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O- and -NH(CH 2 ) 3 O-, the -(CH 2 ) 4 -, -( CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O- and -NH(CH 2 ) 3 O- are optionally substituted with 1, 2 or 3 Rd , and other variables are as defined in the present invention.
  • the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-,- NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O- and -NH(CH 2 ) 3 O-, and other variables are as defined in the present invention.
  • the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
  • R 1 , R 2 , R 3 , R 4 , R 5 and R d are as defined in the present invention.
  • n1 is selected from 3, 4 and 5;
  • n2 is selected from 1, 2 and 3.
  • the present invention also provides the following compounds or their pharmaceutically acceptable salts,
  • the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
  • the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
  • the present invention also provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of medicines for KRAS-related diseases.
  • the compounds of the present invention can be synthesized by the following methods:
  • the lowest energy barrier for the binding of the compound of the present invention in the KRAS G12C protein structure is smaller than the energy barrier of the reference compound AMG510 in the active conformation of the protein structure. Therefore, the compound of the present invention is easier to bind to the protein. In actual binding with the protein, it is possible to exhibit binding activity similar to or better than that of the reference compound AMG510.
  • the compound of the present invention exhibits a good inhibitory activity on the proliferation of Mia PaCa-2 cells, and has a good inhibitory effect on the tumor growth of Mia PaCa-2 xenograft and NCI-H358 xenograft, and the compound of the present invention has excellent pharmacokinetics. Kinetic properties.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues. , Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salt refers to a salt of the compound of the present invention, which is prepared from a compound with specific substituents discovered in the present invention and a relatively non-toxic acid or base.
  • a base addition salt can be obtained by contacting the compound with a sufficient amount of base in a pure solution or a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salt or similar salts.
  • the acid addition salt can be obtained by contacting the compound with a sufficient amount of acid in a pure solution or a suitable inert solvent.
  • Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, the organic acid includes, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid; also include salts of amino acids (such as arginine, etc.) , And salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and
  • the pharmaceutically acceptable salt of the present invention can be synthesized from the parent compound containing acid or base by conventional chemical methods. In general, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of appropriate base or acid in water or organic solvent or a mixture of both.
  • the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers Isomers, (D)-isomers, (L)-isomers, and their racemic mixtures and other mixtures, such as enantiomers or diastereomer-enriched mixtures, all of these mixtures belong to this Within the scope of the invention.
  • Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All these isomers and their mixtures are included in the scope of the present invention.
  • the compound of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms constituting the compound.
  • compounds can be labeled with radioisotopes, such as tritium ( 3 H), iodine-125 ( 125 I), or C-14 ( 14 C).
  • deuterium can be substituted for hydrogen to form deuterated drugs.
  • the bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon.
  • deuterated drugs can reduce toxic side effects and increase drug stability. , Enhance the efficacy, extend the biological half-life of drugs and other advantages. All changes in the isotopic composition of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.
  • substituted means that any one or more hydrogen atoms on a specific atom are replaced by a substituent.
  • the substituent may include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and the compound after substitution Is stable.
  • Oxygen substitution does not occur on aromatic groups.
  • optionally substituted means that it can be substituted or unsubstituted. Unless otherwise specified, the type and number of substituents can be arbitrary on the basis that they can be chemically realized.
  • any variable such as R
  • its definition in each case is independent.
  • the group can optionally be substituted with up to two Rs, and R has independent options in each case.
  • combinations of substituents and/or variants thereof are only permitted if such combinations result in stable compounds.
  • linking group When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
  • the middle linking group L is -MW-, at this time -MW- can be formed by connecting ring A and ring B in the same direction as the reading order from left to right It can also be formed by connecting ring A and ring B in the opposite direction to the reading order from left to right
  • Combinations of the linking groups, substituents, and/or variants thereof are only permitted if such combinations result in stable compounds.
  • any one or more sites of the group can be connected to other groups through chemical bonds.
  • the connection method of the chemical bond is not positioned, and there is a H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will correspondingly decrease with the number of chemical bonds connected to become the corresponding valence number ⁇ The group.
  • the chemical bond between the site and other groups can be a straight solid bond Straight dashed key Or wavy line Express.
  • the straight solid bond in -OCH 3 means that it is connected to other groups through the oxygen atom in the group;
  • the straight dashed bond in indicates that the two ends of the nitrogen atom in the group are connected to other groups;
  • the wavy line in indicates that the phenyl group is connected to other groups through the 1 and 2 carbon atoms;
  • C 1-6 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 6 carbon atoms.
  • the C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 and C 5 alkyl groups, etc.; it may Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine).
  • C 1-6 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl) , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.
  • C 1-4 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 4 carbon atoms.
  • the C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent ( Such as methine).
  • Examples of C 1-4 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl) , S-butyl and t-butyl) and so on.
  • C 1-3 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 3 carbon atoms.
  • the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine) .
  • Examples of C 1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.
  • C n-n+m or C n -C n+m includes any specific case of n to n+m carbons, for example, C 1-12 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , including any range from n to n+m, for example, C 1- 12 includes C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12, etc.; similarly, from n to n +m member means that the number of atoms in the ring is from n to n+m, for example, 3-12 membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those well known to those skilled in the art Equivalent alternatives, preferred implementations include but are not limited to the embodiments of the present invention.
  • the three-dimensional configuration of the compound of the present invention can be determined by test methods such as single crystal diffraction, optical rotation, and CD.
  • the structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the field.
  • SXRD single crystal X-ray diffraction
  • the cultured single crystal is collected with the Bruker D8 venture diffractometer to collect the diffraction intensity data
  • the light source is CuK ⁇ radiation
  • the scanning method After scanning and collecting relevant data, the direct method (Shelxs97) is further used to analyze the crystal structure to confirm the absolute configuration.
  • KOAc stands for potassium acetate
  • Pd(dppf)Cl 2 .DCM stands for [1,1'-bis(diphenylphosphine)ferrocene] dichloropalladium dichloride dichloromethane complex
  • Dioxane stands for 1,4-dioxane
  • DIPEA stands for N,N-diisopropylethylamine
  • DCM stands for dichloromethane
  • Cbz stands for benzyloxycarbonyl
  • iPr stands for isopropyl
  • KHMDS stands for bis(trimethyl) Silyl) potassium amide
  • POCl 3 stands for phosphorus oxychloride
  • CH 3 CN stands for acetonitrile
  • TBS stands for tert-butyldimethylsilyl
  • THF stands for tetrahydrofuran.
  • Figure 1 shows the binding mode of AMG510 and KRAS G12C protein (active conformation: 6IOM).
  • Figure 2 is a 2D diagram of the combination mode of AMG510 and KARS G12C.
  • Figure 3 is a low-energy conformation diagram of AMG510.
  • Figure 4 is a diagram of the rotational dihedral angle and the corresponding energy barrier change of AMG510 (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
  • Figure 5 is a superimposition diagram of the low-energy conformation of WX001 (ie compound 003 of Example 2) and the active conformation of AMG510.
  • Figure 6 shows the dihedral angle of rotation of WX001 (compound 003 of Example 2) and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is isopropyl methylpyridine and Dihedral angle of mother core)
  • Figure 7 is a superimposed view of the low-energy conformation of WX002 and the active conformation of AMG510.
  • Figure 8 shows the rotation dihedral angle of WX002 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
  • Figure 9 is a superimposition of the low-energy conformation of WX003 and the active conformation of AMG510.
  • Figure 10 shows the rotation dihedral angle of WX003 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
  • Figure 11 is a superimposition diagram of the low-energy conformation of WX004 (ie, compound 011 of Example 5) and the active conformation of AMG510.
  • Figure 12 is the rotation dihedral angle of WX004 (ie compound 011 in Example 5) and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is isopropyl methylpyridine and The dihedral angle of the mother core).
  • Figure 13 is a superimposition of the low-energy conformation of WX005 and the active conformation of AMG510.
  • Figure 14 is a diagram of the rotation dihedral angle and the corresponding energy barrier change of WX005 (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
  • Figure 15 is a superimposition diagram of the low-energy conformation of WX006 (ie compound 005 of Example 3) and the active conformation of AMG510.
  • Figure 16 is the rotation dihedral angle of WX006 (ie compound 005 of Example 3) and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is isopropyl methylpyridine and The dihedral angle of the mother core).
  • Figure 17 is a superimposed view of the low-energy conformation of WX007 and the active conformation of AMG510.
  • Figure 18 is a diagram of the rotating dihedral angle and the corresponding energy barrier change of WX007 (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
  • Figure 19 is a superimposed view of the low-energy conformation of WX008 and the active conformation of AMG510.
  • Figure 20 shows the rotation dihedral angle of WX008 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
  • Figure 21 is a superimposed view of the low-energy conformation of WX009 and the active conformation of AMG510.
  • Figure 22 shows the rotation dihedral angle of WX009 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
  • Figure 23 is a superimposed view of the low-energy conformation of WX010 and the low-energy conformation of WX006.
  • Figure 24 is a superimposed view of the low-energy conformation of WX011 and the low-energy conformation of WX006.
  • Figure 25 is a superimposed view of the low-energy conformation of WX012 and the low-energy conformation of WX006.
  • Figure 26 is a superimposed view of the low-energy conformation of WX013 and the low-energy conformation of WX006.
  • Figure 27 is a superimposed view of the low-energy conformation of WX014 and the low-energy conformation of WX006.
  • Figure 28 is a graph showing the tumor growth curve of the Mia PaCa-2 xenograft tumor model tumor-bearing mice after administration of the compound.
  • Figure 29 is a graph showing the body weight change of the Mia PaCa-2 xenograft tumor model tumor-bearing mice during the administration process.
  • Figure 30 is a graph showing the tumor growth curve of NCI-H358 xenograft tumor model tumor-bearing mice after compound administration.
  • Figure 31 is a graph showing the body weight changes of the NCI-H358 xenograft tumor model tumor-bearing mice during the administration process.
  • coordinate1 represents coordinate axis 1
  • coordinate2 represents coordinate axis 2.
  • the dihedral angle (dehidal 1) of fluorophenol and the pyridopyrimidinone core (hereinafter referred to as the core) is 50.6°
  • the dihedral angle (dehidal 2) of isopropyl methylpyridine and the core is 88.3 ° (see Figure 3).
  • AMG510 from its low energy conformation to its protein binding mode active conformation (fluorophenol rotates from 50.6° to 58.8°, isopropyl methylpyridine from 88.3° to 86.8°) needs to overcome the energy barrier of 0.185kcal/mol (see figure) 4).
  • Pyridine fragments are macrocyclic molecules connected by chains, and explored the energy barrier difference between the lowest energy barrier conformation of these macrocyclic molecules and the active conformation in the AMG510 and KRAS G12C protein co-crystals; at the same time, we also used five As an example of meta-linker macrocyclic molecules, we further explored the changes in the lowest energy barrier conformation of these macrocyclic molecules after replacing isopropyl groups with different substituents (methyl, ethyl, propyl, cyclopropyl, tert-butyl, etc.).
  • Dihedral 1 is the dihedral angle of fluorobenzene and fluoropyridine
  • Dihedral 2 is the dihedral angle of isopropylpyridine and pyrimidinone
  • ⁇ E is the active conformation of the protein binding mode from the low energy conformation to AMG510 (Dihedral 1 is 58.8° , Dihedral 2 is 86.8°) the energy barrier that needs to be consumed
  • WX001 is the compound 003 of Example 2
  • WX006 is the compound 005 of Example 3.
  • the low-energy conformation of WX001 ⁇ WX009 overlaps well with the active conformation of AMG510.
  • the lowest energy barrier for the binding of the compound of the present invention in the KRAS G12C protein structure is smaller than the energy barrier of the reference compound AMG510 in the active conformation of the protein structure. Therefore, the compound of the present invention is easier to bind to the protein. In actual binding with the protein, it is possible to exhibit binding activity similar to or better than that of the reference compound AMG510.
  • Dihedral 1 is the dihedral angle of fluorobenzene and fluoropyridine
  • Dihedral 2 is the dihedral angle of picoline and pyrimidinone (WX010) or the dihedral angle of ethylpyridine and pyrimidinone (WX011 ⁇ WX014).
  • reaction system was kept at 100°C and the reaction was stirred for 12 hours.
  • the reaction system is directly concentrated to dryness.
  • compound 001-5 (5.0 g, 23.92 mmol, 1.00 eq) and tetrahydrofuran (75 mL) were added to a pre-dried 250 mL single-necked flask. After the sample was dissolved, the temperature of the system was cooled to 0°C, and a solution of oxalyl chloride (12.15g, 95.69mmol, 8.38mL, 4.00eq) in tetrahydrofuran (15mL) was slowly added dropwise to the system. After the addition, the system was heated to 75°C for 2 hours.
  • compound 001-6 (7.6 g, 18.48 mmol, 1 eq) and tetrahydrofuran (230 mL) were added to a pre-dried 100 mL three-necked flask. After the solution became clear, the system was cooled to -70 ⁇ -60°C, and potassium hexamethyldisilazide (1M tetrahydrofuran solution, 42.50mL, 2.3eq) was slowly added dropwise to the system. After the dripping, the system was warmed to 25°C and stirred at this temperature for 2 hours. Saturated ammonium chloride solution (10 mL) was added to the reaction system to quench the reaction.
  • tetrahydrofuran 40mL was added to the 80mL long tube containing compound 001-8 (1.05g, 2.67mmol, 1.00eq), cooled to 0°C, and N,N'-diisopropylethyl was added to the system Amine (5.18g, 40.05mmol, 6.98mL, 15eq). After the addition, the temperature was controlled to 0°C, and a solution of compound 001-9 (802.14 mg, 4.01 mmol, 1.5 eq) in tetrahydrofuran (20 mL) was slowly added to the system. After the addition, the system was stirred at 25°C for 1 hour.
  • the reaction system was slowly poured into 10 mL of ice water, and 15 mL of ethyl acetate was added to dilute. The system is allowed to stand for liquid separation. After liquid separation, the organic phase was collected, and the aqueous phase was extracted with ethyl acetate (10 mL*1). The combined organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue.
  • Step 8 Synthesis of compound 001-12
  • the aqueous phase was extracted with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to obtain compound 001-13. The crude product is used directly in the next step without purification.
  • Step 11 Synthesis of trifluoroacetate salt of compound 001-15
  • Step 12 Synthesis of compounds 001 and 002
  • Step 3 Synthesis of compounds 003 and 004
  • the product is separated by SFC (Separation method: column: REGIS(s, s) WHELK-O1 (250mm*50mm, 10 ⁇ m); mobile phase: [0.1% ammonia-ethanol]; B (ethanol)%: 55%-55%, 6min), the fractions were concentrated, and deionized water was added to freeze-dry to obtain 003 (ie, compound WX001 of calculation example 1) and 004.
  • SFC Separatation method: column: REGIS(s, s) WHELK-O1 (250mm*50mm, 10 ⁇ m); mobile phase: [0.1% ammonia-ethanol]; B (ethanol)%: 55%-55%, 6min), the fractions were concentrated, and deionized water was added to freeze-dry to obtain 003 (ie, compound WX001 of calculation example 1) and 004.
  • compound 001-12 (0.37g, 584.80 ⁇ mol, 1eq) and compound 4-bromo-1-butene (789.49mg, 5.85mmol, 593.60 ⁇ L, 10eq) were dissolved in acetonitrile (20mL), and potassium carbonate was added (181.86mg, 1.32mmol, 2.25eq) and potassium iodide (189.30mg, 1.14mmol, 1.95eq) were reacted at 80°C for 5 hours.
  • Compound 4-bromo-1-butene (394.74mg, 2.92mmol, 296.80 ⁇ L, 5eq) was added and reacted at 80°C for 5 hours.
  • compound 005-3 (0.24 g, 371.10 ⁇ mol, 1 eq) was dissolved in dichloromethane (24 mL), trifluoroacetic acid (2.12 g, 18.56 mmol, 1.37 mL, 50 eq) was added, and the reaction was carried out at 20° C. for 3 hours.
  • the reaction system was directly concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 005-4.
  • the reaction system was not purified and was directly used in the next reaction.
  • Step 5 Synthesis of compounds 005 and 006
  • compound 005-4 (0.45g, 334.63 ⁇ mol, 1eq, equivalent to 7 trifluoroacetate) was dissolved in dichloromethane (30mL), and N,N'-diisopropylethylamine (518.97) was added. mg, 4.02mmol, 699.42 ⁇ L, 12eq), then add acryloyl chloride (45.43mg, 501.94 ⁇ mol, 40.93 ⁇ L, 1.5eq), and react at -60°C for 0.5 hours. Saturated sodium bicarbonate solution (20 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (20 mL*3) and separated.
  • Step 1 Synthesis of the trifluoroacetate salt of compound 007-1
  • Step 2 Synthesis of compounds 007 and 008 and 009 and 010
  • compound 007-1 (0.19g, 208.91 ⁇ mol, 1eq, converted to 3.2 trifluoroacetate) was dissolved in dichloromethane (20mL), and N,N'-diisopropylethylamine (270.01 mg, 2.09mmol, 363.89 ⁇ L, 10eq), then add acryloyl chloride (28.36mg, 313.37 ⁇ mol, 25.55 ⁇ L, 1.5eq), and react at -60°C for 0.5 hours. A saturated sodium bicarbonate solution (20 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (20 mL*3) and separated.
  • Configuration 1 passed SFC (column: REGIS(s,s) WHELK-O1 (250mm*50mm, 10 ⁇ m); mobile phase: [0.1% ammonia-methanol]; B (methanol)%: 62.5%-62.5%, 3min) After separation and purification, compound 007 (peak position of chiral column: 2.008 min) and compound 008 (peak position of chiral column: 2.516 min) were obtained respectively.
  • Configuration 2 passes SFC (column: REGIS(s, s) WHELK-O1 (250mm*50mm, 10 ⁇ m); mobile phase: [0.1% ammonia-methanol]; B (methanol)%: 50%-50%, 3min) After separation and purification, compound 009 (peak position of chiral column: 1.963 min) and compound 010 (peak position of chiral column: 2.151 min) were obtained respectively.
  • compound 011-1 (8g, 41.45mmol, 1eq) and compound 001-2 (6.58g, 37.31mmol, 0.9eq) were dissolved in N,N-dimethylformamide (120mL), and cesium carbonate ( 27.01g, 82.91mmol, 2eq), react at 60°C for 12 hours.
  • Water 150 mL was added to the reaction system, and the aqueous phase was extracted with methyl tertiary ether (300 mL*2) and separated.
  • the organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product.
  • compound 011-9 (1.1g, 1.59mmol, 1eq) and compound 001-11 (322.53mg, 2.07mmol, 1.3eq) were dissolved in 1,4-dioxane (22mL) and water (5.5mL ), add potassium phosphate (675.52mg, 3.18mmol, 2eq) and 1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (103.71mg, 159.12 ⁇ mol, 0.1eq), React at 80°C for 4 hours.
  • compound 011-10 (1.3 g, 1.70 mmol, 1 eq) was dissolved in tetrahydrofuran (60 mL), tetramethyl ammonium fluoride (789.40 mg, 8.48 mmol, 5 eq) was added, and reacted at 65° C. for 5 hours.
  • the reaction system was directly concentrated under reduced pressure, and then water (20 mL) was added.
  • the aqueous phase was extracted with ethyl acetate (20 mL*4), separated, and the organic phases were combined, washed with saturated brine (20 mL), and dried with anhydrous sodium sulfate. Filter and concentrate under reduced pressure to obtain compound 011-11.
  • the reaction system does not need to be purified and can be used directly in the next reaction.
  • compound 011-12 (1.05 g, 1.65 mmol, 1 eq) was dissolved in dichloromethane (40 mL), trifluoroacetic acid (9.43 g, 82.72 mmol, 6.12 mL, 50 eq) was added, and the reaction was carried out at 30° C. for 2 hours.
  • the reaction system was directly concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 011-13.
  • the reaction system does not need to be purified and can be used directly in the next reaction.
  • compound 011-13 (2g, 1.55mmol, 1eq, equivalent to 6.6 trifluoroacetate) was dissolved in dichloromethane (60mL), and N,N-diisopropylethylamine (3.21g, 24.86mmol) , 4.33mL, 16eq), then add acryloyl chloride (281.28mg, 3.11mmol, 253.40 ⁇ L, 2eq), and react at -60°C for 10 minutes. A saturated sodium bicarbonate solution (50 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (50 mL*3) and separated.
  • the pure product was separated by SFC (column: DAICEL CHIRALCEL OD (250mm*30mm, 10 ⁇ m); mobile phase: [0.1% ammonia-ethanol]; ethanol%: 50%-50%, 15min) and purified to obtain compounds 011 and 012, respectively.
  • the aqueous phase was extracted with ethyl acetate (200 mL*4), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was mixed with the filter cake, and concentrated under reduced pressure to obtain a crude product.
  • Methanol 60 mL was added to the crude product to be slurried for 1 hour, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 013-2.
  • compound 013-5 (1.6g, 2.95mmol, 1eq) and compound 001-11 (689.11mg, 4.42mmol, 1.5eq) were dissolved in dioxane (48mL) and water (12mL), and potassium phosphate was added (1.25g, 5.89mmol, 2eq) and 1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (192.03mg, 294.64 ⁇ mol, 0.1eq), react at 80°C for 5 hours . Water (50 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (60 mL*3) and separated.
  • compound 013-6 0.5g, 808.18 ⁇ mol, 1eq
  • compound 013-7 (2.65g, 16.16mmol, 166.73 ⁇ L, 20eq) were dissolved in acetonitrile (20mL), and potassium iodide (402.47mg, 2.42mmol, 3eq) and potassium carbonate (335.10mg, 2.42mmol, 3eq) at 80°C for 15 hours. Since the reaction was not complete, compound 013-7 (1.33g, 8.08mmol, 10eq), potassium iodide (268.32mg, 1.62mmol, 2eq) and potassium carbonate (223.39mg, 1.62mmol, 2eq) were added and reacted at 80°C for 15 hours.
  • compound 013-8 (0.46g, 669.78 ⁇ mol, 1eq) was dissolved in dichloromethane (70mL), and 1,3-bis(2,4,6-trimethylphenyl)-2-(imidazole Alkylene)(dichlorobenzylidene)(tricyclohexylphosphine)ruthenium (113.73mg, 133.96 ⁇ mol, 0.2eq), reacted at 45°C for 3 hours.
  • the reaction system was directly concentrated under reduced pressure to obtain a crude product.
  • compound 013-10 (0.22g, 332.95 ⁇ mol, 1eq) was dissolved in dichloromethane (9mL), trifluoroacetic acid (1.90g, 16.65mmol, 1.23mL, 50eq) was added and reacted at 20°C for 1 hour.
  • the reaction system was directly concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 013-11.
  • the reaction system does not need to be purified and can be used directly in the next reaction.
  • compound 013-11 (0.3g, 295.06 ⁇ mol, 1eq, equivalent to 4 trifluoroacetate) was dissolved in dichloromethane (15mL), and diisopropylethylamine (610.15mg, 4.72mmol, 822.30) ⁇ L, 16eq), then add acryloyl chloride (53.41mg, 590.13 ⁇ mol, 48.12 ⁇ L, 2eq), and react at -60°C for 10 minutes. Saturated sodium bicarbonate solution (20 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (50 mL*3) and separated.
  • the pure product was separated by SFC (column: DAICEL CHIRALPAK IG (250mm*30mm, 10 ⁇ m); mobile phase: [0.1% ammonia-ethanol]; ethanol%: 55%-55%, min) and purified to obtain compound 013 and compound 015, respectively , Compound 016 and Mixture A.
  • Mixture A was separated again by SFC (column: DAICEL CHIRALPAK IG (250mm*30mm, 10 ⁇ m); mobile phase: [0.1% ammonia-ethanol]; ethanol%: 45%-45%, min), and purified to obtain compound 014 and compound 013 .
  • DMEM medium fetal bovine serum was purchased from Biosera, and horse serum was purchased from Gibco.
  • CellTiter-Glo cell viability chemiluminescence detection reagent
  • MIA-PA-CA-2 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd. EnVision multi-label analyzer (PerkinElmer).
  • MIA-PA-CA-2 cells were planted in a white 96-well plate, 80 ⁇ L of cell suspension per well, which contained 1000 MIA-PA-CA-2 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture.
  • the compound to be tested is diluted 5-fold to the 8th concentration with a discharge gun, that is, diluted from 2mM to 26nM, and a double-well experiment is set up.
  • the concentration of the compound transferred to the cell plate ranges from 10 ⁇ M to 0.13 nM.
  • the cell plate was placed in a carbon dioxide incubator for 3 days. Another cell plate is prepared, and the signal value is read as the maximum value (Max value in the following equation) on the day of drug addition to participate in data analysis.
  • Add 50 ⁇ L of cell viability chemiluminescence detection reagent to each well of this cell plate, and incubate for 10 minutes at room temperature to stabilize the luminescence signal. Use multi-marker analyzer to read.
  • IC 50 can be obtained by curve fitting with four parameters ("log(" in GraphPad Prism software) Inhibitor)/Response-Variable Scope" model derived).
  • Table 3 provides the inhibitory activity of the compounds of the present invention on the proliferation of MIA-PA-CA-2 cells.
  • the compound of the present invention exhibits a good inhibitory activity on the proliferation of MIA-PA-CA-2 cells, and the compound 005 (ie WX006 of Calculation Example 1) has an IC 50 of inhibitory activity on MIA-PA-CA-2 cells. nM, significantly better than the in vitro activity of AMG510.
  • This result is consistent with the conclusion drawn by computational chemistry method in Calculation Example 1 (the actual binding of WX006 , a compound of the same structure with a lower ⁇ E energy barrier, to KRAS G12C protein may exhibit similar or better results than the reference compound AMG510.
  • the binding activity is highly consistent, which fully demonstrates the accuracy of the computational chemistry method in the present invention. Therefore, compounds with a lower ⁇ E energy barrier predicted by the computational chemistry method of the present invention will exhibit similar or better inhibitory activity in cell proliferation than the reference compound AMG510.
  • Test Example 2 In vivo pharmacokinetic study
  • the test compound was mixed with a 10% dimethyl sulfoxide/60% polyethylene glycol 400/30% aqueous solution, vortexed and sonicated to prepare a 1 mg/mL clear solution, which was filtered with a microporous membrane for use.
  • Select male SD mice aged 7 to 10 weeks, two in each group.
  • the candidate compound solution was administered intravenously, the dose of AMG510 was 3 mg/kg, and the dose of compound 003 was 2 mg/kg.
  • the candidate compound solution was administered orally at a dose of 10 mg/kg. Collect whole blood for a certain period of time, prepare plasma, analyze drug concentration by LC-MS/MS method, and use Phoenix WinNonlin software (Pharsight, USA) to calculate pharmacokinetic parameters.
  • Table 4 The experimental results are shown in Table 4:
  • Cell culture Human pancreatic cancer Mia PaCa-2 cells (ATCC-CRL-1420) are cultured in a monolayer in vitro, and the culture conditions are DMEM/F12 medium with 20% fetal bovine serum, 1% double antibody, 37°C and 5% carbon dioxide incubation Box culture. Use pancreatin-EDTA for routine digestion and passage twice a week. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, and resuspended in an appropriate amount of PBS. Matrigel is added 1:1 to obtain a cell suspension with a cell density of 25 ⁇ 10 6 cells/mL .
  • Mia PaCa-2 cells (with matrigel, volume ratio 1:1) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached 190mm At 3 o'clock, random grouping was performed according to tumor volume, and the administration was started according to the schedule in Table 5.
  • PO oral administration
  • QD once a day
  • the tumor diameter was measured with vernier calipers twice a week.
  • the anti-tumor efficacy of the compound was evaluated by TGI (%) or the relative tumor growth rate T/C (%).
  • Relative tumor proliferation rate T/C(%) TRTV/CRTV ⁇ 100% (TRTV: treatment group RTV; CRTV: negative control group RTV).
  • the relative tumor volume (RTV) is calculated according to the results of tumor measurement.
  • the average tumor volume, TRTV and CRTV data on the same day.
  • TGI (%) reflects the tumor growth inhibition rate.
  • TGI(%) [(1-(Average tumor volume at the end of a certain treatment group-average tumor volume at the start of the treatment group))/(Average tumor volume at the end of treatment in the solvent control group-start treatment in the solvent control group Average tumor volume at time)] ⁇ 100%.
  • the tumor volume of the tumor-bearing mice in the solvent control group reached 1572 mm 3 after administration for 21 days, and the tumor volume of the test substances compound 003 (5 mg/kg), compound 003 (10 mg/kg) and compound 003 (30 mg/kg)
  • the average values are 46mm 3 , 111mm 3 and 14mm 3 ; T/C are 2.79%, 8.27% and 0.90% respectively; TGI are 110.37%, 105.70% and 112.71%, respectively. All three concentrations can significantly inhibit tumor growth. (P values are all less than 0.001).
  • the average tumor volume of the test substances AMG510 (5mg/kg) and AMG510 (10mg/kg) were 229mm 3 and 86mm 3 , respectively, the T/C was 14.90% and 5.81%, the TGI was 97.17% and 107.51%, and the p values were all ⁇ 0.001, also has a significant anti-tumor effect, and the weight of each dose group of mice is stable, and there is no obvious intolerance.
  • Cell culture Human non-small cell lung cancer NCI-H358 is cultured in a monolayer in vitro.
  • the culture conditions are DMEM/F12 medium plus 20% fetal bovine serum, 1% double antibody, 37°C and 5% carbon dioxide incubator.
  • the cells are collected, counted, and resuspended in an appropriate amount of PBS.
  • Matrigel is added 1:1 to obtain a cell suspension with a cell density of 25 ⁇ 10 6 cells/mL .
  • NCI-H358 cells (with matrigel, volume ratio 1:1) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached 100- At 150 mm 3 , random grouping was performed according to the tumor volume, and the administration was started according to the schedule in Table 6.
  • PO oral administration
  • QD once a day
  • the tumor diameter was measured with vernier calipers twice a week.
  • the anti-tumor efficacy of the compound was evaluated by TGI (%) or the relative tumor growth rate T/C (%).
  • Relative tumor proliferation rate T/C(%) TRTV/CRTV ⁇ 100% (TRTV: treatment group RTV; CRTV: negative control group RTV).
  • the relative tumor volume (RTV) is calculated according to the results of tumor measurement.
  • the average tumor volume, TRTV and CRTV data on the same day.
  • TGI (%) reflects the tumor growth inhibition rate.
  • TGI(%) [(1-(Average tumor volume at the end of a certain treatment group-average tumor volume at the start of the treatment group))/(Average tumor volume at the end of treatment in the solvent control group-start treatment in the solvent control group Average tumor volume at time)] ⁇ 100%.
  • the volume averages are 307mm 3 , 138mm 3 and 50mm 3 respectively; T/C are 55%, 24% and 9% respectively; TGI are 54%, 91% and 111%, respectively. All three concentrations can significantly inhibit tumors Growth (all p values are less than 0.001).
  • the average tumor volume of the test substance AMG510 (5mg/kg) is 248mm 3 , the T/C is 44%, the TGI is 68%, and the p values are all ⁇ 0.001. It also has a significant tumor suppressor effect.
  • the weight is stable and there is no obvious intolerance.

Abstract

Provided are a series of macrocyclic compounds and application thereof in preparation of drugs for KRAS inhibitor-related diseases. Specifically provided are a compound represented by formula (I) and a pharmaceutically acceptable salt thereof.

Description

作为KRAS抑制剂的大环类化合物Macrocyclic compounds as KRAS inhibitors
本发明主张如下优先权：The present invention claims the following priority:
CN202010071528.3，申请日：2020年01月21日；CN202010071528.3, application date: January 21, 2020;
CN202010514548.3，申请日：2020年06月08日；CN202010514548.3, application date: June 08, 2020;
CN202010986192.3，申请日：2020年09月18日。CN202010986192.3, application date: September 18, 2020.
技术领域Technical field
本发明涉及一系列大环类化合物，及其在制备与KRAS抑制剂相关疾病的药物中的应用。具体涉及式(I)所示化合物及其药学上可接受的盐。The present invention relates to a series of macrocyclic compounds and their application in the preparation of drugs for diseases related to KRAS inhibitors. Specifically, it relates to a compound represented by formula (I) and a pharmaceutically acceptable salt thereof.
背景技术Background technique
第一个RAS癌基因发现自大鼠肉瘤(rat sarcoma)，因此得名。RAS蛋白是由RAS基因表达的产物，指一类紧密相关的，由189个氨基酸组成的单体球蛋白，其分子量为21KDa。RAS蛋白可以与鸟嘌呤三核苷酸磷酸(GTP)或鸟嘌呤二核苷酸磷酸(GDP)结合，RAS蛋白的活性状态对细胞的生长、分化、细胞骨架、蛋白质运输和分泌等都具有影响，其活性是通过与GTP或GDP的结合进行调节：当RAS蛋白与GDP结合时，它处于休眠状态，也就是“失活”状态；当有上游特定的细胞生长因子刺激时，RAS蛋白被诱导交换GDP，与GTP结合，此时称为“活化”状态。与GTP结合的RAS蛋白能够活化下游的蛋白，进行信号传递。RAS蛋白自身具有弱的水解GTP水解活性，能够水解GTP到GDP。这样就可以实现从活化状态到失活状态的转化。在这个水解过程中，还需要GAP(GTPase activating proteins,GTP水解酶活化蛋白)参与。它能与RAS蛋白作用，大大促进其水解GTP到GDP的能力。RAS蛋白的突变将影响其与GAP的作用，也就影响了其水解GTP到GDP的能力，使其一直处于活化状态。活化的RAS蛋白持续的给予下游蛋白生长信号，最终导致细胞不停的生长和分化，最终产生肿瘤。RAS基因家族成员众多，其中与各种癌症密切相关的亚家族主要有克尔斯滕大鼠肉瘤病毒致癌基因同源物(KRAS)、哈维大鼠肉瘤病毒致癌同源物(HRAS)和神经母细胞瘤大鼠肉瘤病毒致癌基因同源物(NRAS)。人们发现大约30％的人类肿瘤中都携带某些突变的RAS基因，其中以KRAS突变最为显著，占到所有RAS突变中的86％。对于KRAS突变，最为常见的突变出现在12号甘氨酸(G12)，13号甘氨酸(G13)和61号谷氨酰胺(Q61)残基上，其中G12突变占到83％。The first RAS oncogene was found in rat sarcoma (rat sarcoma), hence the name. The RAS protein is a product expressed by the RAS gene, which refers to a closely related monomer globulin consisting of 189 amino acids with a molecular weight of 21KDa. RAS protein can be combined with guanine trinucleotide phosphate (GTP) or guanine dinucleotide phosphate (GDP). The active state of RAS protein has an impact on cell growth, differentiation, cytoskeleton, protein transport and secretion, etc. Its activity is regulated by binding to GTP or GDP: when the RAS protein binds to GDP, it is in a dormant state, that is, "inactive"; when stimulated by a specific upstream cell growth factor, the RAS protein is induced Exchange GDP and combine with GTP, which is called "activated" state at this time. The RAS protein bound to GTP can activate downstream proteins for signal transmission. RAS protein itself has weak GTP hydrolysis activity and can hydrolyze GTP to GDP. In this way, the transformation from the activated state to the inactivated state can be realized. In this hydrolysis process, GAP (GTPase activating proteins) is also required to participate. It can interact with RAS protein and greatly promote its ability to hydrolyze GTP to GDP. The mutation of RAS protein will affect its interaction with GAP, which will also affect its ability to hydrolyze GTP to GDP, making it always in an activated state. The activated RAS protein continuously gives growth signals to downstream proteins, which eventually leads to the continuous growth and differentiation of cells, and eventually tumors. There are many members of the RAS gene family, among which the subfamilies closely related to various cancers are mainly Kirsten rat sarcoma virus oncogene homolog (KRAS), Harvey rat sarcoma virus carcinogenic homolog (HRAS) and nerve The blastoma rat sarcoma virus oncogene homolog (NRAS). It has been found that about 30% of human tumors carry certain mutated RAS genes, of which KRAS mutations are the most significant, accounting for 86% of all RAS mutations. For KRAS mutations, the most common mutations appear on glycine 12 (G12), glycine 13 (G13) and glutamine (Q61) residues, of which G12 mutation accounts for 83%.
G12C突变是KRAS基因突变中比较常见的一个亚型，它是指12号甘氨酸突变为半胱氨酸。KRAS G12C突变在肺癌中最为常见，根据文献(Nat Rev Drug Discov 2014；13:828-851)报道的数据推算，KRAS G12C突变占到所有肺癌患者的10％左右。 G12C mutation is a more common subtype of KRAS gene mutation, which refers to the mutation of glycine 12 to cysteine. KRAS G12C mutation is the most common in lung cancer. According to data reported in the literature (Nat Rev Drug Discov 2014; 13:828-851), KRAS G12C mutation accounts for about 10% of all lung cancer patients.
KRAS G12C突变蛋白作为一个前沿靶点，目前的研究还不是很多。 KRAS G12C mutant protein is a cutting-edge target, and there are not many studies at present.
文献J Med Chem.2020 Jan 9；6 3(1):52-65报道了AMG510(结构如下)与KRAS G12C蛋白的共晶结构(ID:6IOM)。根据文献报道，AMG510结合在KRAS G12C蛋白的SwichⅡ口袋中，丙烯基和Cys12加 成形成共价键，羰基与Lys16形成氢键，母核嘧啶酮并吡啶分别和Tyr96形成pi-pi作用，异丙基甲基吡啶和母核之间以86.8°的二面角***疏水口袋，氟苯酚和母核以58.8°的二面角***疏水口袋，同时酚羟基和Arg68形成氢键作用。(见图1和图2，Maestro 2017-2，Pymol 1.8.6)。 The document J Med Chem. 2020 Jan 9; 63(1): 52-65 reported the co-crystal structure (ID: 6IOM) of AMG510 (structure as follows) and KRAS G12C protein. According to reports in the literature, AMG510 binds to the SwichⅡ pocket of KRAS G12C protein. The propylene group and Cys12 form a covalent bond. The carbonyl group forms a hydrogen bond with Lys16. The parent pyrimidinone and pyridine form a pi-pi interaction with Tyr96, respectively. The pyridine and the mother core are inserted into the hydrophobic pocket with a dihedral angle of 86.8°, and the fluorophenol and the mother core are inserted into the hydrophobic pocket with a dihedral angle of 58.8°. At the same time, the phenolic hydroxyl group and Arg68 form a hydrogen bond. (See Figure 1 and Figure 2, Maestro 2017-2, Pymol 1.8.6).
Figure PCTCN2021073149-appb-000001
Figure PCTCN2021073149-appb-000001
发明内容Summary of the invention
本发明提供了式(I)所示化合物或其药学上可接受的盐，The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
Figure PCTCN2021073149-appb-000002
Figure PCTCN2021073149-appb-000002
其中，in,
R 1选自H、F、Cl、Br、I和CH 3，所述CH 3任选被1、2或3个R a取代； R 1 is selected H, F, Cl, Br, I and CH 3, CH 3 optionally substituted by the two or three R a;
R 2选自H、F、Cl、Br和I； R 2 is selected from H, F, Cl, Br and I;
R 3选自H、F、Cl、Br、I和C 1-3烷基，所述C 1-3烷基任选被1、2或3个R b取代； R 3 is selected from H, F, Cl, Br, and C 1-3 alkyl, said C 1-3 alkyl optionally substituted with 1, 2 or 3 R b;
R 4选自H、F、Cl、Br和I； R 4 is selected from H, F, Cl, Br and I;
R 5选自C 1-6烷基和环丙基，所述C 1-6烷基和环丙基任选被1、2或3个R c取代； R 5 is selected from C 1-6 alkyl and cyclopropyl, the C 1-6 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c ;
L 1选自-(CH 2) m-、-(CH 2) m-NR 6-、-(CH 2) m-O-、-NR 6-(CH 2) m-NR 6-、-NR 6-(CH 2) m-O-、-O-(CH 2) m-O-和-(CH 2) n-(CH＝CH) p-(CH 2) m-O-，所述-(CH 2) m-、-(CH 2) m-NR 6-、-(CH 2) m-O-、-NR 6-(CH 2) m-NR 6-、-NR 6-(CH 2) m-O-、-O-(CH 2) m-O-和-(CH 2) n-(CH＝CH) p-(CH 2) m-O-任选被1、2或3个R d取代； L 1 is selected from -(CH 2 ) m -, -(CH 2 ) m -NR 6 -, -(CH 2 ) m -O-, -NR 6 -(CH 2 ) m -NR 6 -, -NR 6 -(CH 2 ) m -O-, -O-(CH 2 ) m -O- and -(CH 2 ) n -(CH=CH) p -(CH 2 ) m -O-, the -(CH 2 ) m -, -(CH 2 ) m -NR 6 -, -(CH 2 ) m -O-, -NR 6 -(CH 2 ) m -NR 6 -, -NR 6 -(CH 2 ) m- O-, -O-(CH 2 ) m -O- and -(CH 2 ) n -(CH=CH) p -(CH 2 ) m -O- are optionally substituted by 1, 2 or 3 R d ;
m选自1、2、3、4、5和6；m is selected from 1, 2, 3, 4, 5 and 6;
n选自0和1；n is selected from 0 and 1;
p选自1和2；p is selected from 1 and 2;
R 6选自H和CH 3； R 6 is selected from H and CH 3 ;
各R a、R b和R c分别独立地选自H、F、Cl、Br和I； Each R a, R b and R c are each independently selected from H, F, Cl, Br and I;
R d选自H、F、Cl、Br、I和CH 3； R d is selected from H, F, Cl, Br, I and CH 3 ;
带“*”碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在。The carbon atom with "*" is a chiral carbon atom and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其选自In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
Figure PCTCN2021073149-appb-000003
Figure PCTCN2021073149-appb-000003
其中，R 1、R 2、R 3、R 4、R 5和L 1如本发明所定义。 Wherein, R 1 , R 2 , R 3 , R 4 , R 5 and L 1 are as defined in the present invention.
在本发明的一些方案中，上述R 1选自H，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 1 is selected from H, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 2选自H，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 2 is selected from H, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 3选自CH 3，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 3 is selected from CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 4选自F，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 4 is selected from F, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自C 1-4烷基和环丙基，所述C 1-4烷基和环丙基任选被1、2或3个R c取代，其他变量如本发明所定义。 In some embodiments of the present invention, the above-mentioned R 5 is selected from C 1-4 alkyl and cyclopropyl, the C 1-4 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c , and others The variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000004
所述CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000005
任选被1、2或3个R c取代，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000004
The CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000005
Optionally substituted by 1, 2 or 3 R c , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000006
其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000006
Other variables are as defined in the present invention.
在本发明的一些方案中，上述m选自1、2、3、4和5，其他变量如本发明所定义。In some aspects of the present invention, the aforementioned m is selected from 1, 2, 3, 4, and 5, and other variables are as defined in the present invention.
在本发明的一些方案中，上述L 1选自-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-、-(CH＝CH)-(CH 2) 2-O-和-(CH＝CH)-(CH 2) 3-O-，所述-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-、-(CH＝CH)-(CH 2) 2-O-和-(CH＝CH)-(CH 2) 3-O-任选被1、2或3个R d取代，其他变量如本发明所定义。 In some aspects of the present invention, the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O (CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH =CH)-CH 2 -O-, -(CH=CH)-(CH 2 ) 2 -O- and -(CH=CH)-(CH 2 ) 3 -O-, the -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-, -NH(CH 2 ) 2 O-,- (CH 2 ) 4 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH=CH)-CH 2 -O-, -(CH=CH)-(CH 2 ) 2 -O- and -(CH=CH)-(CH 2 ) 3 -O- are optionally substituted by 1, 2 or 3 Rd , and other variables are as defined in the present invention.
在本发明的一些方案中，上述L 1选自-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-、-(CH＝CH)-(CH 2) 2-O-和-(CH 2) 3CH(CH 3)O-，其他变量如本发明所定义。 In some aspects of the present invention, the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O (CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH =CH)-CH 2 -O-, -(CH=CH)-(CH 2 ) 2 -O- and -(CH 2 ) 3 CH(CH 3 )O-, and other variables are as defined in the present invention.
本发明还提供了式(I)所示化合物或其药学上可接受的盐，The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
Figure PCTCN2021073149-appb-000007
Figure PCTCN2021073149-appb-000007
其中，in,
R 1选自H、F、Cl、Br、I和CH 3，所述CH 3任选被1、2或3个R a取代； R 1 is selected H, F, Cl, Br, I and CH 3, CH 3 optionally substituted by the two or three R a;
R 2选自H、F、Cl、Br和I； R 2 is selected from H, F, Cl, Br and I;
R 3选自H、F、Cl、Br、I和C 1-3烷基，所述C 1-3烷基任选被1、2或3个R b取代； R 3 is selected from H, F, Cl, Br, and C 1-3 alkyl, said C 1-3 alkyl optionally substituted with 1, 2 or 3 R b;
R 4选自H、F、Cl、Br和I； R 4 is selected from H, F, Cl, Br and I;
R 5选自C 1-6烷基和环丙基，所述C 1-6烷基和环丙基任选被1、2或3个R c取代； R 5 is selected from C 1-6 alkyl and cyclopropyl, the C 1-6 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c ;
L 1选自-(CH 2) m-、-(CH 2) m-NR 6-、-(CH 2) m-O-、-NR 6-(CH 2) m-NR 6-、-NR 6-(CH 2) m-O-、-O-(CH 2) m-O-和-(CH 2) n-(CH＝CH) p-(CH 2) m-O-，所述-(CH 2) m-、-(CH 2) m-NR 6-、-(CH 2) m-O-、-NR 6-(CH 2) m-NR 6-、-NR 6-(CH 2) m-O-、-O-(CH 2) m-O-和-(CH 2) n-(CH＝CH) p-(CH 2) m-O-任选被1、2或3个R d取代； L 1 is selected from -(CH 2 ) m -, -(CH 2 ) m -NR 6 -, -(CH 2 ) m -O-, -NR 6 -(CH 2 ) m -NR 6 -, -NR 6 -(CH 2 ) m -O-, -O-(CH 2 ) m -O- and -(CH 2 ) n -(CH=CH) p -(CH 2 ) m -O-, the -(CH 2 ) m -, -(CH 2 ) m -NR 6 -, -(CH 2 ) m -O-, -NR 6 -(CH 2 ) m -NR 6 -, -NR 6 -(CH 2 ) m- O-, -O-(CH 2 ) m -O- and -(CH 2 ) n -(CH=CH) p -(CH 2 ) m -O- are optionally substituted by 1, 2 or 3 R d ;
m选自1、2、3、4、5和6；m is selected from 1, 2, 3, 4, 5 and 6;
n选自0和1；n is selected from 0 and 1;
p选自1和2；p is selected from 1 and 2;
R 6选自H和CH 3； R 6 is selected from H and CH 3 ;
各R a、R b、R c和R d分别独立地选自H、F、Cl、Br和I； Each R a, R b, R c and R d are each independently selected from H, F, Cl, Br and I;
带“*”碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在。The carbon atom with "*" is a chiral carbon atom and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其选自In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
Figure PCTCN2021073149-appb-000008
Figure PCTCN2021073149-appb-000008
其中，R 1、R 2、R 3、R 4、R 5和L 1如本发明所定义。 Wherein, R 1 , R 2 , R 3 , R 4 , R 5 and L 1 are as defined in the present invention.
在本发明的一些方案中，上述R 1选自H，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 1 is selected from H, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 2选自H，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 2 is selected from H, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 3选自CH 3，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 3 is selected from CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 4选自F，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 4 is selected from F, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自C 1-4烷基和环丙基，所述C 1-4烷基和环丙基任选被1、2或3个R c取代，其他变量如本发明所定义。 In some embodiments of the present invention, the above-mentioned R 5 is selected from C 1-4 alkyl and cyclopropyl, the C 1-4 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c , and others The variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000009
所述CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000010
任选被1、2或3个R c取代，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000009
The CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000010
Optionally substituted by 1, 2 or 3 R c , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000011
其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000011
Other variables are as defined in the present invention.
在本发明的一些方案中，上述m选自1、2、3和4，其他变量如本发明所定义。In some aspects of the present invention, the aforementioned m is selected from 1, 2, 3, and 4, and other variables are as defined in the present invention.
在本发明的一些方案中，上述L 1选自-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-、-(CH＝CH)-(CH 2) 2-O-和-(CH＝CH)-(CH 2) 3-O-，所述-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-(CH 2) 2-O-和任选被1、2或3个R d取代，其他变量如本发明所定义。 In some aspects of the present invention, the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O (CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH =CH)-CH 2 -O-, -(CH=CH)-(CH 2 ) 2 -O- and -(CH=CH)-(CH 2 ) 3 -O-, the -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-, -NH(CH 2 ) 2 O-,- (CH 2 ) 4 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH=CH)-(CH 2 ) 2 -O- and optionally by 1, 2 Or 3 R d substitutions, and other variables are as defined in the present invention.
在本发明的一些方案中，上述L 1选自-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-和-(CH＝CH)-(CH 2) 2-O-，其他变量如本发明所定义。 In some aspects of the present invention, the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O (CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH =CH)-CH 2 -O- and -(CH=CH)-(CH 2 ) 2 -O-, other variables are as defined in the present invention.
本发明提供了式(I)所示化合物或其药学上可接受的盐，The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
Figure PCTCN2021073149-appb-000012
Figure PCTCN2021073149-appb-000012
其中，in,
R 1选自H、F、Cl、Br、I和CH 3，所述CH 3任选被1、2或3个R a取代； R 1 is selected H, F, Cl, Br, I and CH 3, CH 3 optionally substituted by the two or three R a;
R 2选自H、F、Cl、Br和I； R 2 is selected from H, F, Cl, Br and I;
R 3选自H、F、Cl、Br、I和C 1-3烷基，所述C 1-3烷基任选被1、2或3个R b取代； R 3 is selected from H, F, Cl, Br, and C 1-3 alkyl, said C 1-3 alkyl optionally substituted with 1, 2 or 3 R b;
R 4选自H、F、Cl、Br和I； R 4 is selected from H, F, Cl, Br and I;
R 5选自C 1-6烷基和环丙基，所述C 1-6烷基和环丙基任选被1、2或3个R c取代； R 5 is selected from C 1-6 alkyl and cyclopropyl, the C 1-6 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c ;
L 1选自-(CH 2) m-、-(CH 2) m-NH-、-(CH 2) m-O-、-NH-(CH 2) m-NH-、-NH-(CH 2) m-O-和-O-(CH 2) m-O-，所述-(CH 2) m-、-(CH 2) m-NH-、-(CH 2) m-O-、-NH-(CH 2) m-NH-、-NH-(CH 2) m-O-和-O-(CH 2) m-O-任选被1、2或3个R d取代； L 1 is selected from -(CH 2 ) m -, -(CH 2 ) m -NH-, -(CH 2 ) m -O-, -NH-(CH 2 ) m -NH-, -NH-(CH 2 ) m -O- and -O-(CH 2 ) m -O-, the -(CH 2 ) m -, -(CH 2 ) m -NH-, -(CH 2 ) m -O-, -NH -(CH 2 ) m -NH-, -NH-(CH 2 ) m -O- and -O-(CH 2 ) m -O- are optionally substituted by 1, 2 or 3 R d ;
m选自2、3、4、5和6；m is selected from 2, 3, 4, 5 and 6;
各R a、R b、R c和R d分别独立地选自H、F、Cl、Br和I； Each R a, R b, R c and R d are each independently selected from H, F, Cl, Br and I;
带“*”碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在。The carbon atom with "*" is a chiral carbon atom and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其选自In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
Figure PCTCN2021073149-appb-000013
Figure PCTCN2021073149-appb-000013
其中，R 1、R 2、R 3、R 4、R 5和L 1如本发明所定义。 Wherein, R 1 , R 2 , R 3 , R 4 , R 5 and L 1 are as defined in the present invention.
在本发明的一些方案中，上述R 1选自H，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 1 is selected from H, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 2选自H，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 2 is selected from H, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 3选自CH 3，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 3 is selected from CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 4选自F，其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 4 is selected from F, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自C 1-4烷基和环丙基，所述C 1-4烷基和环丙基任选被1、2或3个R c取代，其他变量如本发明所定义。 In some embodiments of the present invention, the above-mentioned R 5 is selected from C 1-4 alkyl and cyclopropyl, the C 1-4 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c , and others The variables are as defined in the present invention.
在本发明的一些方案中，上述R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000014
所述CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000015
任选被1、2或3个R c取代。 In some aspects of the present invention, the above-mentioned R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000014
The CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000015
Optionally substituted with 1, 2 or 3 R c .
在本发明的一些方案中，上述R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
Figure PCTCN2021073149-appb-000016
其他变量如本发明所定义。 In some aspects of the present invention, the above-mentioned R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
Figure PCTCN2021073149-appb-000016
Other variables are as defined in the present invention.
在本发明的一些方案中，上述m选自2、3和4，其他变量如本发明所定义。In some aspects of the present invention, the above m is selected from 2, 3 and 4, and other variables are as defined in the present invention.
在本发明的一些方案中，上述L 1选自-(CH 2) 4-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-和-NH(CH 2) 3O-，所述-(CH 2) 4-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-和-NH(CH 2) 3O-任选被1、2或3个R d取代，其他变量如本发明所定义。 In some aspects of the present invention, the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-,- NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O- and -NH(CH 2 ) 3 O-, the -(CH 2 ) 4 -, -( CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O- and -NH(CH 2 ) 3 O- are optionally substituted with 1, 2 or 3 Rd , and other variables are as defined in the present invention.
在本发明的一些方案中，上述L 1选自-(CH 2) 4-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-(CH 2) 4O-、-O(CH 2) 3O-和-NH(CH 2) 3O-，其他变量如本发明所定义。 In some aspects of the present invention, the aforementioned L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-,- NH(CH 2 ) 2 O-, -(CH 2 ) 4 O-, -O(CH 2 ) 3 O- and -NH(CH 2 ) 3 O-, and other variables are as defined in the present invention.
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其选自In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
Figure PCTCN2021073149-appb-000017
Figure PCTCN2021073149-appb-000017
Figure PCTCN2021073149-appb-000018
Figure PCTCN2021073149-appb-000018
其中，in,
R 1、R 2、R 3、R 4、R 5和R d如本发明所定义； R 1 , R 2 , R 3 , R 4 , R 5 and R d are as defined in the present invention;
m1选自3、4和5；m1 is selected from 3, 4 and 5;
m2选自1、2和3。m2 is selected from 1, 2 and 3.
本发明还有一些方案是由上述变量任意组合而来。There are also some schemes of the present invention that come from any combination of the above variables.
本发明还提供了下列所示化合物或其药学上可接受的盐，The present invention also provides the following compounds or their pharmaceutically acceptable salts,
Figure PCTCN2021073149-appb-000019
Figure PCTCN2021073149-appb-000019
Figure PCTCN2021073149-appb-000020
Figure PCTCN2021073149-appb-000020
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其选自In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
Figure PCTCN2021073149-appb-000021
Figure PCTCN2021073149-appb-000021
Figure PCTCN2021073149-appb-000022
Figure PCTCN2021073149-appb-000022
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其选自In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from
Figure PCTCN2021073149-appb-000023
Figure PCTCN2021073149-appb-000023
本发明还提供了上述化合物或其药学上可接受的盐在制备与KRAS相关疾病的药物中的应用。The present invention also provides the use of the above-mentioned compound or its pharmaceutically acceptable salt in the preparation of medicines for KRAS-related diseases.
本发明化合物可以通过下述方法合成：The compounds of the present invention can be synthesized by the following methods:
方法1：method 1:
Figure PCTCN2021073149-appb-000024
Figure PCTCN2021073149-appb-000024
方法2：Method 2:
Figure PCTCN2021073149-appb-000025
Figure PCTCN2021073149-appb-000025
方法3：Method 3:
Figure PCTCN2021073149-appb-000026
Figure PCTCN2021073149-appb-000026
方法4：Method 4:
Figure PCTCN2021073149-appb-000027
Figure PCTCN2021073149-appb-000027
技术效果Technical effect
本发明化合物在KRAS G12C蛋白结构中的结合最低能垒，与参照化合物AMG510在该蛋白结构中的活性构象下的能垒，具有较小的能量差异，因此本发明化合物较容易与该蛋白结合，在与该蛋白的实际结合中有可能展现与参照化合物AMG510相似的或更优的结合活性。本发明化合物对Mia PaCa-2细胞增殖展现出较好的抑制活性，对Mia PaCa-2异种移植和NCI-H358异种移植的肿瘤生长有较好的抑制作用，且本发明化合物具有优异的药代动力学性质。 The lowest energy barrier for the binding of the compound of the present invention in the KRAS G12C protein structure is smaller than the energy barrier of the reference compound AMG510 in the active conformation of the protein structure. Therefore, the compound of the present invention is easier to bind to the protein. In actual binding with the protein, it is possible to exhibit binding activity similar to or better than that of the reference compound AMG510. The compound of the present invention exhibits a good inhibitory activity on the proliferation of Mia PaCa-2 cells, and has a good inhibitory effect on the tumor growth of Mia PaCa-2 xenograft and NCI-H358 xenograft, and the compound of the present invention has excellent pharmacokinetics. Kinetic properties.
相关定义Related definitions
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered uncertain or unclear without a special definition, but should be understood in its ordinary meaning. When a trade name appears in this article, it is meant to refer to its corresponding commodity or its active ingredient.
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and/or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues. , Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机胺或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，所述无机酸包括例如盐酸、氢溴酸、硝酸、碳酸，碳酸氢根，磷酸、磷酸一氢根、磷酸二氢根、硫酸、硫酸氢根、氢碘酸、亚磷酸等；以及有机酸盐，所述有机酸包括如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸和甲磺酸等类似的酸；还包括氨基酸(如精氨酸等)的盐，以及如葡糖醛酸等有机酸的盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。The term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention, which is prepared from a compound with specific substituents discovered in the present invention and a relatively non-toxic acid or base. When the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salt or similar salts. When the compound of the present invention contains a relatively basic functional group, the acid addition salt can be obtained by contacting the compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, the organic acid includes, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid; also include salts of amino acids (such as arginine, etc.) , And salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups, which can be converted into any base or acid addition salt.
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。The pharmaceutically acceptable salt of the present invention can be synthesized from the parent compound containing acid or base by conventional chemical methods. In general, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of appropriate base or acid in water or organic solvent or a mixture of both.
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers Isomers, (D)-isomers, (L)-isomers, and their racemic mixtures and other mixtures, such as enantiomers or diastereomer-enriched mixtures, all of these mixtures belong to this Within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All these isomers and their mixtures are included in the scope of the present invention.
本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚( 3H)，碘-125( 125I)或C-14( 14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。 The compound of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms constituting the compound. For example, compounds can be labeled with radioisotopes, such as tritium ( 3 H), iodine-125 ( 125 I), or C-14 ( 14 C). For another example, deuterium can be substituted for hydrogen to form deuterated drugs. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , Enhance the efficacy, extend the biological half-life of drugs and other advantages. All changes in the isotopic composition of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.
术语“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。The term "optional" or "optionally" refers to the event or condition described later that may but not necessarily occur, and the description includes a situation in which the event or condition occurs and a situation in which the event or condition does not occur .
术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，取代基可以包括重氢和氢的变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧(即＝O)时，意 味着两个氢原子被取代。氧取代不会发生在芳香基上。术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent. The substituent may include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and the compound after substitution Is stable. When the substituent is oxygen (i.e. =0), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it can be substituted or unsubstituted. Unless otherwise specified, the type and number of substituents can be arbitrary on the basis that they can be chemically realized.
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。When any variable (such as R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group can optionally be substituted with up to two Rs, and R has independent options in each case. In addition, combinations of substituents and/or variants thereof are only permitted if such combinations result in stable compounds.
当一个连接基团的数量为0时，比如-(CRR) 0-，表示该连接基团为单键。 When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
当其中一个变量选自单键时，表示其连接的两个基团直接相连，比如A-L-Z中L代表单键时表示该结构实际上是A-Z。When one of the variables is selected from a single bond, it means that the two connected groups are directly connected. For example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.
当所列举的连接基团没有指明其连接方向，其连接方向是任意的，例如，
Figure PCTCN2021073149-appb-000028
中连接基团L为-M-W-，此时-M-W-既可以按与从左往右的读取顺序相同的方向连接环A和环B构成
Figure PCTCN2021073149-appb-000029
也可以按照与从左往右的读取顺序相反的方向连接环A和环B构成
Figure PCTCN2021073149-appb-000030
所述连接基团、取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。 When the listed linking group does not indicate its linking direction, its linking direction is arbitrary, for example,
Figure PCTCN2021073149-appb-000028
The middle linking group L is -MW-, at this time -MW- can be formed by connecting ring A and ring B in the same direction as the reading order from left to right
Figure PCTCN2021073149-appb-000029
It can also be formed by connecting ring A and ring B in the opposite direction to the reading order from left to right
Figure PCTCN2021073149-appb-000030
Combinations of the linking groups, substituents, and/or variants thereof are only permitted if such combinations result in stable compounds.
除非另有规定，当某一基团具有一个或多个可连接位点时，该基团的任意一个或多个位点可以通过化学键与其他基团相连。当该化学键的连接方式是不定位的，且可连接位点存在H原子时，则连接化学键时，该位点的H原子的个数会随所连接化学键的个数而对应减少变成相应价数的基团。所述位点与其他基团连接的化学键可以用直形实线键
Figure PCTCN2021073149-appb-000031
直形虚线键
Figure PCTCN2021073149-appb-000032
或波浪线
Figure PCTCN2021073149-appb-000033
表示。例如-OCH 3中的直形实线键表示通过该基团中的氧原子与其他基团相连；
Figure PCTCN2021073149-appb-000034
中的直形虚线键表示通过该基团中的氮原子的两端与其他基团相连；
Figure PCTCN2021073149-appb-000035
中的波浪线表示通过该苯基基团中的1和2位碳原子与其他基团相连；
Figure PCTCN2021073149-appb-000036
表示该哌啶基上的任意可连接位点可以通过1个化学键与其他基团相连，至少包括
Figure PCTCN2021073149-appb-000037
这4种连接方式，即使-N-上画出 了H原子，但是
Figure PCTCN2021073149-appb-000038
仍包括
Figure PCTCN2021073149-appb-000039
这种连接方式的基团，只是在连接1个化学键时，该位点的的H会对应减少1个变成相应的一价哌啶基。 Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the connection method of the chemical bond is not positioned, and there is a H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will correspondingly decrease with the number of chemical bonds connected to become the corresponding valence number的组。 The group. The chemical bond between the site and other groups can be a straight solid bond
Figure PCTCN2021073149-appb-000031
Straight dashed key
Figure PCTCN2021073149-appb-000032
Or wavy line
Figure PCTCN2021073149-appb-000033
Express. For example , the straight solid bond in -OCH 3 means that it is connected to other groups through the oxygen atom in the group;
Figure PCTCN2021073149-appb-000034
The straight dashed bond in indicates that the two ends of the nitrogen atom in the group are connected to other groups;
Figure PCTCN2021073149-appb-000035
The wavy line in indicates that the phenyl group is connected to other groups through the 1 and 2 carbon atoms;
Figure PCTCN2021073149-appb-000036
Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least
Figure PCTCN2021073149-appb-000037
These four connection methods, even though the H atom is drawn on -N-, but
Figure PCTCN2021073149-appb-000038
Still include
Figure PCTCN2021073149-appb-000039
The group in this connection mode, only when one chemical bond is connected, the H at this position will decrease by one and become the corresponding monovalent piperidinyl group.
除非另有规定，术语“C 1-6烷基”用于表示直链或支链的由1至6个碳原子组成的饱和碳氢基团。所述C 1-6烷基包括C 1-5、C 1-4、C 1-3、C 1-2、C 2-6、C 2-4、C 6和C 5烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-6烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)、戊基(包括n-戊基，异戊基和新戊基)、己基等。 Unless otherwise specified, the term "C 1-6 alkyl" is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 6 carbon atoms. The C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 and C 5 alkyl groups, etc.; it may Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine). Examples of C 1-6 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl) , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.
除非另有规定，术语“C 1-4烷基”用于表示直链或支链的由1至4个碳原子组成的饱和碳氢基团。所述C 1-4烷基包括C 1-2、C 1-3和C 2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-4烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)等。 Unless otherwise specified, the term "C 1-4 alkyl" is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 4 carbon atoms. The C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent ( Such as methine). Examples of C 1-4 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl) , S-butyl and t-butyl) and so on.
除非另有规定，术语“C 1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C 1-3烷基包括C 1-2和C 2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-3烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)等。 Unless otherwise specified, the term "C 1-3 alkyl" is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 3 carbon atoms. The C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine) . Examples of C 1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.
除非另有规定，C n-n+m或C n-C n+m包括n至n+m个碳的任何一种具体情况，例如C 1-12包括C 1、C 2、C 3、C 4、C 5、C 6、C 7、C 8、C 9、C 10、C 11、和C 12，也包括n至n+m中的任何一个范围，例如C 1- 12包括C 1-3、C 1-6、C 1-9、C 3-6、C 3-9、C 3-12、C 6-9、C 6-12、和C 9-12等；同理，n元至n+m元表示环上原子数为n至n+m个，例如3-12元环包括3元环、4元环、5元环、6元环、7元环、8元环、9元环、10元环、11元环、和12元环，也包括n至n+m中的任何一个范围，例如3-12元环包括3-6元环、3-9元环、5-6元环、5-7元环、6-7元环、6-8元环、和6-10元环等。 Unless otherwise specified, C n-n+m or C n -C n+m includes any specific case of n to n+m carbons, for example, C 1-12 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , including any range from n to n+m, for example, C 1- 12 includes C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12, etc.; similarly, from n to n +m member means that the number of atoms in the ring is from n to n+m, for example, 3-12 membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring , 10-membered ring, 11-membered ring, and 12-membered ring, including any range from n to n+m, for example, 3-12 membered ring includes 3-6 membered ring, 3-9 membered ring, 5-6 membered ring Ring, 5-7 membered ring, 6-7 membered ring, 6-8 membered ring, 6-10 membered ring, etc.
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those well known to those skilled in the art Equivalent alternatives, preferred implementations include but are not limited to the embodiments of the present invention.
本发明的化合物存在立体构型的，可以通过单晶衍射、旋光、CD等试验方法确定。The three-dimensional configuration of the compound of the present invention can be determined by test methods such as single crystal diffraction, optical rotation, and CD.
本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：
Figure PCTCN2021073149-appb-000040
扫描，收集相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。 The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the field. For example, single crystal X-ray diffraction (SXRD), the cultured single crystal is collected with the Bruker D8 venture diffractometer to collect the diffraction intensity data, the light source is CuKα radiation, and the scanning method:
Figure PCTCN2021073149-appb-000040
After scanning and collecting relevant data, the direct method (Shelxs97) is further used to analyze the crystal structure to confirm the absolute configuration.
本发明所使用的溶剂可经市售获得。本发明采用下述缩略词：KOAc代表醋酸钾；Pd(dppf)Cl 2.DCM代表[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物；Dioxane代表1,4-二氧六环；DIPEA代表N,N-二异丙基乙胺；DCM代表二氯甲烷；Cbz代表苄氧羰基；iPr代表异丙基；KHMDS代表双(三甲基硅烷基)氨基钾；POCl 3代表三氯氧磷；CH 3CN代表乙腈；TBS代表叔丁基二甲基甲硅烷基；THF 代表四氢呋喃。 The solvent used in the present invention is commercially available. The present invention uses the following abbreviations: KOAc stands for potassium acetate; Pd(dppf)Cl 2 .DCM stands for [1,1'-bis(diphenylphosphine)ferrocene] dichloropalladium dichloride dichloromethane complex ; Dioxane stands for 1,4-dioxane; DIPEA stands for N,N-diisopropylethylamine; DCM stands for dichloromethane; Cbz stands for benzyloxycarbonyl; iPr stands for isopropyl; KHMDS stands for bis(trimethyl) Silyl) potassium amide; POCl 3 stands for phosphorus oxychloride; CH 3 CN stands for acetonitrile; TBS stands for tert-butyldimethylsilyl; THF stands for tetrahydrofuran.
化合物依据本领域常规命名原则或者使用
Figure PCTCN2021073149-appb-000041
软件命名，市售化合物采用供应商目录名称。 Compounds are based on conventional naming principles in the field or use
Figure PCTCN2021073149-appb-000041
The software is named, and the commercially available compounds use the supplier catalog name.
附图说明Description of the drawings
图1为AMG510和KRAS G12C蛋白的结合模式(活性构象：6IOM)。 Figure 1 shows the binding mode of AMG510 and KRAS G12C protein (active conformation: 6IOM).
图2为AMG510和KARS G12C的结合模式2D图。 Figure 2 is a 2D diagram of the combination mode of AMG510 and KARS G12C.
图3为AMG510的低能构象图。Figure 3 is a low-energy conformation diagram of AMG510.
图4为AMG510的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 4 is a diagram of the rotational dihedral angle and the corresponding energy barrier change of AMG510 (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
图5为WX001(即实施例2的化合物003)的低能构象和AMG510活性构象叠合图。Figure 5 is a superimposition diagram of the low-energy conformation of WX001 (ie compound 003 of Example 2) and the active conformation of AMG510.
图6为WX001(即实施例2的化合物003)的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)Figure 6 shows the dihedral angle of rotation of WX001 (compound 003 of Example 2) and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is isopropyl methylpyridine and Dihedral angle of mother core)
图7为WX002的低能构象和AMG510活性构象叠合图。Figure 7 is a superimposed view of the low-energy conformation of WX002 and the active conformation of AMG510.
图8为WX002的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 8 shows the rotation dihedral angle of WX002 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
图9为WX003的低能构象和AMG510活性构象叠合图。Figure 9 is a superimposition of the low-energy conformation of WX003 and the active conformation of AMG510.
图10为WX003的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 10 shows the rotation dihedral angle of WX003 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
图11为WX004(即实施例5的化合物011)的低能构象和AMG510活性构象叠合图。Figure 11 is a superimposition diagram of the low-energy conformation of WX004 (ie, compound 011 of Example 5) and the active conformation of AMG510.
图12为WX004(即实施例5的化合物011)的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 12 is the rotation dihedral angle of WX004 (ie compound 011 in Example 5) and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is isopropyl methylpyridine and The dihedral angle of the mother core).
图13为WX005的低能构象和AMG510活性构象叠合图。Figure 13 is a superimposition of the low-energy conformation of WX005 and the active conformation of AMG510.
图14为WX005的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 14 is a diagram of the rotation dihedral angle and the corresponding energy barrier change of WX005 (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
图15为WX006(即实施例3的化合物005)的低能构象和AMG510活性构象叠合图。Figure 15 is a superimposition diagram of the low-energy conformation of WX006 (ie compound 005 of Example 3) and the active conformation of AMG510.
图16为WX006(即实施例3的化合物005)的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 16 is the rotation dihedral angle of WX006 (ie compound 005 of Example 3) and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is isopropyl methylpyridine and The dihedral angle of the mother core).
图17为WX007的低能构象和AMG510活性构象叠合图。Figure 17 is a superimposed view of the low-energy conformation of WX007 and the active conformation of AMG510.
图18为WX007的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 18 is a diagram of the rotating dihedral angle and the corresponding energy barrier change of WX007 (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
图19为WX008的低能构象和AMG510活性构象叠合图。Figure 19 is a superimposed view of the low-energy conformation of WX008 and the active conformation of AMG510.
图20为WX008的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 20 shows the rotation dihedral angle of WX008 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
图21为WX009的低能构象和AMG510活性构象叠合图。Figure 21 is a superimposed view of the low-energy conformation of WX009 and the active conformation of AMG510.
图22为WX009的旋转二面角以及相对应的能垒变化图(坐标1为氟苯酚与母核的二面角，坐标2为异丙基甲基吡啶与母核的二面角)。Figure 22 shows the rotation dihedral angle of WX009 and the corresponding energy barrier change diagram (coordinate 1 is the dihedral angle between fluorophenol and the core, and coordinate 2 is the dihedral angle between isopropyl methylpyridine and the core).
图23为WX010的低能构象和WX006低能构象叠合图。Figure 23 is a superimposed view of the low-energy conformation of WX010 and the low-energy conformation of WX006.
图24为WX011的低能构象和WX006低能构象叠合图。Figure 24 is a superimposed view of the low-energy conformation of WX011 and the low-energy conformation of WX006.
图25为WX012的低能构象和WX006低能构象叠合图。Figure 25 is a superimposed view of the low-energy conformation of WX012 and the low-energy conformation of WX006.
图26为WX013的低能构象和WX006低能构象叠合图。Figure 26 is a superimposed view of the low-energy conformation of WX013 and the low-energy conformation of WX006.
图27为WX014的低能构象和WX006低能构象叠合图。Figure 27 is a superimposed view of the low-energy conformation of WX014 and the low-energy conformation of WX006.
图28为Mia PaCa-2异种移植瘤模型荷瘤鼠在给予化合物后的肿瘤生长曲线图。Figure 28 is a graph showing the tumor growth curve of the Mia PaCa-2 xenograft tumor model tumor-bearing mice after administration of the compound.
图29为Mia PaCa-2异种移植瘤模型荷瘤鼠在给药过程中的体重变化图。Figure 29 is a graph showing the body weight change of the Mia PaCa-2 xenograft tumor model tumor-bearing mice during the administration process.
图30为NCI-H358异种移植瘤模型荷瘤鼠在给予化合物后的肿瘤生长曲线图。Figure 30 is a graph showing the tumor growth curve of NCI-H358 xenograft tumor model tumor-bearing mice after compound administration.
图31为NCI-H358异种移植瘤模型荷瘤鼠在给药过程中的体重变化图。Figure 31 is a graph showing the body weight changes of the NCI-H358 xenograft tumor model tumor-bearing mice during the administration process.
注：coordinate1代表坐标轴1；coordinate2代表坐标轴2。Note: coordinate1 represents coordinate axis 1; coordinate2 represents coordinate axis 2.
具体实施方式Detailed ways
下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。The present invention will be described in detail through the following examples, but it is not meant to impose any disadvantageous restriction on the present invention. The present invention has been described in detail herein, and its specific embodiments are also disclosed. For those skilled in the art, various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. Will be obvious.
参考例5Reference example 5
Figure PCTCN2021073149-appb-000042
Figure PCTCN2021073149-appb-000042
通过薛定谔公司Maestro软件的Macromodel模块计算AMG510的低能构象。在低能构象中氟苯酚与吡啶并嘧啶酮母核(以下简称为母核)的二面角(dehidal 1)为50.6°，异丙基甲基吡啶与母核二面角(dehidal 2)为88.3°(见图3)。AMG510从其低能构象变换到其蛋白结合模式活性构象(氟苯酚从50.6°旋转到58.8°，异丙基甲基吡啶从88.3°旋转到86.8°)需要克服0.185kcal/mol的能垒(见图4)。Calculate the low-energy conformation of AMG510 through the Macromodel module of Schrodinger Maestro software. In the low-energy conformation, the dihedral angle (dehidal 1) of fluorophenol and the pyridopyrimidinone core (hereinafter referred to as the core) is 50.6°, and the dihedral angle (dehidal 2) of isopropyl methylpyridine and the core is 88.3 ° (see Figure 3). The conversion of AMG510 from its low energy conformation to its protein binding mode active conformation (fluorophenol rotates from 50.6° to 58.8°, isopropyl methylpyridine from 88.3° to 86.8°) needs to overcome the energy barrier of 0.185kcal/mol (see figure) 4).
计算例1Calculation example 1
Figure PCTCN2021073149-appb-000043
Figure PCTCN2021073149-appb-000043
通过观察AMG510和KRAS G12C蛋白的活性构象结合模式，我们发现氟苯酚片段和异丙基甲基吡 啶片段在自由旋转的同时，与吡啶并嘧啶酮母核间形成的最低能垒构象(低能构象)，与AMG510和KRAS G12C蛋白共晶中的蛋白结合模式构象(活性构象)较为吻合，这解释了AMG510和KRAS G12C蛋白的高结合活性。一般地，小分子的最低能垒构象(低能构象)与其在该蛋白中的结合模式构象(活性构象)间的能垒差越小，意味着该小分子从低能构象变换到与该蛋白的结合活性构象时损失的能量就越小，化合物越容易与蛋白相结合，其结合活性将较高。 By observing the active conformational binding mode of AMG510 and KRAS G12C protein, we found that the fluorophenol fragment and isopropyl picoline fragment form the lowest energy barrier conformation (low energy conformation) with the pyridopyrimidinone nucleus while rotating freely. , Which is more consistent with the protein binding mode conformation (active conformation) in the co-crystal of AMG510 and KRAS G12C protein, which explains the high binding activity of AMG510 and KRAS G12C protein. Generally, the smaller the energy barrier between the lowest energy barrier conformation (low energy conformation) of a small molecule and the binding mode conformation (active conformation) of the protein, the smaller the energy barrier difference, which means that the small molecule changes from the low energy conformation to the binding of the protein. The less energy is lost in the active conformation, the easier it is for the compound to bind to the protein, and its binding activity will be higher.
为了锁定AMG510的活性构象，进一步降低其旋转能垒，我们通过不同长度的连接链将AMG510的氟苯酚片段和异丙基甲基吡啶片段环合，得到了一系列不同的氟苯酚片段和异丙基甲基吡啶片段以链相连接的大环分子，并探索了这些大环分子的最低能垒构象与AMG510和KRAS G12C蛋白共晶中的活性构象间的能垒差别；同时，我们也以五元linker大环分子为例，进一步探索了不同取代基(甲基、乙基、丙基、环丙基、叔丁基等)替代异丙基后，这些大环分子最低能垒构象的变化。 In order to lock the active conformation of AMG510 and further reduce its rotational energy barrier, we cyclized the fluorophenol fragment and isopropylmethylpyridine fragment of AMG510 through different length linking chains to obtain a series of different fluorophenol fragments and isopropylmethylpyridine fragments. Pyridine fragments are macrocyclic molecules connected by chains, and explored the energy barrier difference between the lowest energy barrier conformation of these macrocyclic molecules and the active conformation in the AMG510 and KRAS G12C protein co-crystals; at the same time, we also used five As an example of meta-linker macrocyclic molecules, we further explored the changes in the lowest energy barrier conformation of these macrocyclic molecules after replacing isopropyl groups with different substituents (methyl, ethyl, propyl, cyclopropyl, tert-butyl, etc.).
经Macromodel模块计算WX001～WX009的低能构象旋转二面角和旋转能垒，结果如表1所示。The low-energy conformational rotation dihedral angle and rotation energy barrier of WX001～WX009 were calculated by Macromodel module, and the results are shown in Table 1.
表1.本发明化合物低能构象旋转二面角和旋转能垒Table 1. Low-energy conformational rotation dihedral angle and rotation energy barrier of the compounds of the present invention
化合物Compound Dihedral1(度)Dihedral1 (degrees) Dihedral2(度)Dihedral2 (degrees) ΔE(Kcal/mol)ΔE(Kcal/mol)
AMG510(活性构象)AMG510 (active conformation) 58.858.8 86.886.8 //
AMG510(低能构象)AMG510 (low energy conformation) 50.650.6 88.388.3 0.1850.185
WX001(低能构象)WX001 (low energy conformation) 54.754.7 98.898.8 0.7690.769
WX002(低能构象)WX002 (low energy conformation) 60.660.6 96.496.4 0.4340.434
WX003(低能构象)WX003 (low energy conformation) 52.452.4 97.497.4 0.7270.727
WX004(低能构象)WX004 (low energy conformation) 56.256.2 102.3102.3 1.1391.139
WX005(低能构象)WX005 (low energy conformation) 56.256.2 103.2103.2 1.3981.398
WX006(低能构象)WX006 (low energy conformation) 59.259.2 90.890.8 0.0030.003
WX007(低能构象)WX007 (low energy conformation) 6969 92.692.6 0.2560.256
WX008(低能构象)WX008 (low energy conformation) 67.867.8 9595 0.5310.531
WX009(低能构象)WX009 (low energy conformation) 66.366.3 88.388.3 0.2090.209
注：Dihedral 1为氟苯和氟吡啶的二面角，Dihedral 2为异丙基吡啶和嘧啶酮的二面角，ΔE为从低能构象变换到AMG510的蛋白结合模式活性构象(Dihedral 1为58.8°，Dihedral 2为86.8°)需要消耗的能垒；WX001即实施例2的化合物003，WX006即实施例3的化合物005。Note: Dihedral 1 is the dihedral angle of fluorobenzene and fluoropyridine, Dihedral 2 is the dihedral angle of isopropylpyridine and pyrimidinone, and ΔE is the active conformation of the protein binding mode from the low energy conformation to AMG510 (Dihedral 1 is 58.8° , Dihedral 2 is 86.8°) the energy barrier that needs to be consumed; WX001 is the compound 003 of Example 2 and WX006 is the compound 005 of Example 3.
结论：WX001～WX009的低能构象与AMG510的活性构象叠合好。本发明化合物在KRAS G12C蛋白结构中的结合最低能垒，与参照化合物AMG510在该蛋白结构中的活性构象下的能垒，具有较小 的能量差异，因此本发明化合物较容易与该蛋白结合，在与该蛋白的实际结合中有可能展现与参照化合物AMG510相似的或更优的结合活性。 Conclusion: The low-energy conformation of WX001～WX009 overlaps well with the active conformation of AMG510. The lowest energy barrier for the binding of the compound of the present invention in the KRAS G12C protein structure is smaller than the energy barrier of the reference compound AMG510 in the active conformation of the protein structure. Therefore, the compound of the present invention is easier to bind to the protein. In actual binding with the protein, it is possible to exhibit binding activity similar to or better than that of the reference compound AMG510.
经Macromodel模块计算WX010～WX014的低能构象旋转二面角，结果如表2所示。The low-energy conformational rotation dihedral angles of WX010～WX014 were calculated by the Macromodel module, and the results are shown in Table 2.
表2.本发明化合物低能构象旋转二面角Table 2. Low-energy conformational rotation dihedral angles of the compounds of this invention
化合物Compound Dihedral1(度)Dihedral1 (degrees) Dihedral2(度)Dihedral2 (degrees)
WX010(低能构象)WX010 (low energy conformation) 59.659.6 91.191.1
WX011(低能构象)WX011 (low energy conformation) 59.559.5 91.691.6
WX012(低能构象)WX012 (low energy conformation) 59.859.8 87.887.8
WX013(低能构象)WX013 (low energy conformation) 60.060.0 90.890.8
WX014(低能构象)WX014 (low energy conformation) 61.761.7 89.689.6
注：Dihedral 1为氟苯和氟吡啶的二面角，Dihedral 2为甲基吡啶和嘧啶酮的二面角(WX010)或乙基吡啶和嘧啶酮的二面角(WX011～WX014)。Note: Dihedral 1 is the dihedral angle of fluorobenzene and fluoropyridine, and Dihedral 2 is the dihedral angle of picoline and pyrimidinone (WX010) or the dihedral angle of ethylpyridine and pyrimidinone (WX011～WX014).
结论：WX010～WX014的低能构象与WX006的低能构象基本一致，在与KRAS G12C蛋白的实际结合中有可能展现与参照化合物AMG510相似的或更优的结合活性。 Conclusion: The low-energy conformation of WX010～WX014 is basically the same as that of WX006 . In actual binding with KRAS G12C protein, it is possible to exhibit binding activity similar to or better than that of the reference compound AMG510.
实施例1Example 1
Figure PCTCN2021073149-appb-000044
Figure PCTCN2021073149-appb-000044
合成路线：synthetic route:
Figure PCTCN2021073149-appb-000045
Figure PCTCN2021073149-appb-000045
步骤1：化合物001-2的合成Step 1: Synthesis of compound 001-2
在预先干燥过的500mL三口瓶中加入化合物001-1(20g,122.70mmol,1.00eq)，异丙烯基三氟硼酸钾(19.06g,128.83mmol,1.05eq)，碳酸钾(25.44g,184.04mmol,1.5eq),1,4-二氧六环(200mL)和水(20mL)。加完后，置换氮气三次。然后体系中加入1,1'-双(二苯基膦基)二茂铁]二氯化钯(1.09g,1.49mmol,1.22e-2eq)。加完后，反应体系在100℃，反应搅拌12小时。反应体系直接浓缩干。粗品用硅胶柱层析(石油醚：乙酸乙酯＝8:1至5:1)，得到化合物001-2。Add compound 001-1 (20g, 122.70mmol, 1.00eq), potassium isopropenyl trifluoroborate (19.06g, 128.83mmol, 1.05eq), potassium carbonate (25.44g, 184.04mmol) into a pre-dried 500mL three-necked flask. , 1.5eq), 1,4-dioxane (200mL) and water (20mL). After the addition, replace with nitrogen three times. Then 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.09g, 1.49mmol, 1.22e-2eq) was added to the system. After the addition, the reaction system was kept at 100°C and the reaction was stirred for 12 hours. The reaction system is directly concentrated to dryness. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 8:1 to 5:1) to obtain compound 001-2.
1H NMR(400MHz,CDCl 3)δ＝7.82(d,J＝5.2Hz,1H),7.01(d,J＝5.1Hz,1H),5.43(s,1H),5.25(s,1H),4.28(br s,2H),2.09(s,3H)。 1 H NMR(400MHz,CDCl 3 )δ=7.82(d,J=5.2Hz,1H), 7.01(d,J=5.1Hz,1H), 5.43(s,1H), 5.25(s,1H), 4.28 (br s, 2H), 2.09 (s, 3H).
步骤2：化合物001-3的合成Step 2: Synthesis of compound 001-3
向预先用氮气置换过的500mL单口瓶加入三(三苯基膦)氯化铑(3.10g,3.35mmol,3.65e-2eq)。加完后，将甲醇(300mL)和化合物001-2(15.5g,91.92mmol,1eq)加入到体系中。加料完毕，将体系用氢气置换3次。体系在25℃搅拌1.0小时。将反应体系过滤，滤液浓缩。粗产品通过快速柱层析分离(石油醚:乙酸乙酯＝5:1至3:1)，纯化得到化合物001-3。Tris(triphenylphosphine) rhodium chloride (3.10g, 3.35mmol, 3.65e-2eq) was added to a 500mL single-necked flask which had been replaced with nitrogen in advance. After the addition, methanol (300 mL) and compound 001-2 (15.5 g, 91.92 mmol, 1 eq) were added to the system. After the feeding was completed, the system was replaced with hydrogen three times. The system was stirred at 25°C for 1.0 hour. The reaction system was filtered, and the filtrate was concentrated. The crude product was separated by flash column chromatography (petroleum ether: ethyl acetate = 5:1 to 3:1), and purified to obtain compound 001-3.
LCMS:MS m/z:171.2[M+1] +。 LCMS: MS m/z: 171.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝7.82(d,J＝5.2Hz,1H),6.96(d,J＝5.2Hz,1H),4.03(br s,2H),3.01-2.88(m,1H),1.22(d,J＝6.8Hz,6H)。 1 H NMR (400MHz, CDCl 3 ) δ = 7.82 (d, J = 5.2 Hz, 1H), 6.96 (d, J = 5.2 Hz, 1H), 4.03 (br s, 2H), 3.01-2.88 (m, 1H) ), 1.22 (d, J=6.8 Hz, 6H).
步骤3：化合物001-4的合成Step 3: Synthesis of compound 001-4
在预先干燥过的250mL三口瓶中加入化合物001-3(10g,58.60mmol,1.00eq)和N,N-二甲基甲酰胺(100mL)。加完后，置换氮气三次。然后体系中加入化合物烯丙基三丁基锡(58.93g,177.98mmol,54.57mL,3.04eq)和1,1'-双(二苯基膦基)二茂铁]二氯化钯(4.29g,5.86mmol,0.10eq)。加完后，置换氮气三次。反应体系在135℃，反应搅拌12小时。将反应体系降温到室温后，再向体系中加入预先配置好的饱和氟化钾溶液(250mL)淬灭反应。再向体系中加入水(150mL)和乙酸乙酯萃取(150mL*2)。合并有机相，无水硫酸钠干燥，过滤，滤液浓缩。粗产品通过快速柱层析分离(石油醚:乙酸乙酯＝10:1至8:1)，纯化得到化合物001-4。Compound 001-3 (10 g, 58.60 mmol, 1.00 eq) and N,N-dimethylformamide (100 mL) were added to a pre-dried 250 mL three-necked flask. After the addition, replace with nitrogen three times. Then the compound allyl tributyl tin (58.93g, 177.98mmol, 54.57mL, 3.04eq) and 1,1'-bis(diphenylphosphino)ferrocene) palladium dichloride (4.29g, 5.86) were added to the system mmol, 0.10eq). After the addition, replace with nitrogen three times. The reaction system was kept at 135°C and the reaction was stirred for 12 hours. After the reaction system was cooled to room temperature, the pre-prepared saturated potassium fluoride solution (250 mL) was added to the system to quench the reaction. Water (150 mL) and ethyl acetate were added to the system for extraction (150 mL*2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was separated by flash column chromatography (petroleum ether: ethyl acetate=10:1 to 8:1), and purified to obtain compound 001-4.
LCMS:MS(ESI)m/z:177.1[M+1] +。 LCMS: MS (ESI) m/z: 177.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝7.97(d,J＝4.8Hz,1H),6.81(d,J＝4.8Hz,1H),5.97-5.83(m,1H),5.19-5.04(m,2H),3.66(br s,2H),3.26(d,J＝6.4Hz,2H),3.08-2.94(m,1H),1.28(d,J＝6.8Hz,6H)。 1 H NMR(400MHz, CDCl 3 )δ=7.97(d,J=4.8Hz,1H), 6.81(d,J=4.8Hz,1H), 5.97-5.83(m,1H), 5.19-5.04(m, 2H), 3.66 (br s, 2H), 3.26 (d, J=6.4 Hz, 2H), 3.08-2.94 (m, 1H), 1.28 (d, J=6.8 Hz, 6H).
步骤4：化合物001-6的合成Step 4: Synthesis of compound 001-6
氮气保护，向预先干燥好的250mL单口瓶中加入化合物001-5(5.0g,23.92mmol,1.00eq)和四氢呋喃(75mL)。待样品溶解后，将体系温度冷却到0℃，向体系中缓慢滴加草酰氯(12.15g,95.69mmol,8.38mL,4.00eq)四氢呋喃(15mL)溶液。加完后，将体系升温至75℃反应2小时。随后将反应体系直接减压浓缩，0℃下向反应体系中加入四氢呋喃(75mL)。然后0℃下，缓慢滴加化合物001-4(4.09g,23.21mmol,0.97eq)的四氢呋喃(15mL)溶液，0℃下反应2小时。向反应体系中加入饱和氯化铵溶液(25mL)淬灭反应。接着使用乙酸乙酯(50mL*3)萃取，分液。合并有机相，使用无水硫酸钠干燥，过滤，减压浓缩得到化合物001-6。粗产品未纯化直接用于下一步。Under nitrogen protection, compound 001-5 (5.0 g, 23.92 mmol, 1.00 eq) and tetrahydrofuran (75 mL) were added to a pre-dried 250 mL single-necked flask. After the sample was dissolved, the temperature of the system was cooled to 0°C, and a solution of oxalyl chloride (12.15g, 95.69mmol, 8.38mL, 4.00eq) in tetrahydrofuran (15mL) was slowly added dropwise to the system. After the addition, the system was heated to 75°C for 2 hours. Subsequently, the reaction system was directly concentrated under reduced pressure, and tetrahydrofuran (75 mL) was added to the reaction system at 0°C. Then, at 0°C, a solution of compound 001-4 (4.09g, 23.21mmol, 0.97eq) in tetrahydrofuran (15mL) was slowly added dropwise, and reacted at 0°C for 2 hours. Saturated ammonium chloride solution (25 mL) was added to the reaction system to quench the reaction. Then, it was extracted with ethyl acetate (50 mL*3) and separated. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 001-6. The crude product was used directly in the next step without purification.
LCMS:MS(ESI)m/z:411.0[M+1] +。 LCMS: MS (ESI) m/z: 411.0 [M+1] + .
步骤5：化合物001-7的合成Step 5: Synthesis of compound 001-7
氮气保护，向预先干燥好的100mL三口瓶中加入化合物001-6(7.6g,18.48mmol,1eq)和四氢呋喃(230mL)。等溶液变澄清后，将体系冷却到-70～-60℃下，向体系中缓慢滴加六甲基二硅基胺基钾(1M的四氢呋喃溶液,42.50mL,2.3eq)。滴加完后，体系回温到25℃并在此温度下搅拌2小时。向反应体系中加入饱和氯化铵溶液(10mL)淬灭反应。接着使用乙酸乙酯(10mL*3)萃取，分液。合并有 机相，使用饱和食盐水(10mL)清洗，分液，有机相使用无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过柱层析(洗脱液：石油醚：乙酸乙酯＝20:1到8:1)分离纯化得到化合物001-7。Under nitrogen protection, compound 001-6 (7.6 g, 18.48 mmol, 1 eq) and tetrahydrofuran (230 mL) were added to a pre-dried 100 mL three-necked flask. After the solution became clear, the system was cooled to -70～-60°C, and potassium hexamethyldisilazide (1M tetrahydrofuran solution, 42.50mL, 2.3eq) was slowly added dropwise to the system. After the dripping, the system was warmed to 25°C and stirred at this temperature for 2 hours. Saturated ammonium chloride solution (10 mL) was added to the reaction system to quench the reaction. Then, it was extracted with ethyl acetate (10 mL*3) and separated. The organic phases were combined, washed with saturated brine (10 mL), and separated. The organic phase was dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography (eluent: petroleum ether: ethyl acetate = 20:1 to 8:1) to obtain compound 001-7.
LCMS:MS(ESI)m/z:375.0[M+1] +。 LCMS: MS (ESI) m/z: 375.0 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.91(br s,1H),8.58(d,J＝5.2Hz,1H),8.19(d,J＝6.4Hz,1H),7.29(d,J＝4.8Hz,1H),6.47-6.29(m,1H),5.90(dd,J＝1.2,15.6Hz,1H),2.69-2.56(m,1H),1.74(dd,J＝1.6,6.4Hz,3H),1.15(d,J＝6.8Hz,3H),1.06(d,J＝6.8Hz,3H)。 1 H NMR (400MHz, CDCl 3 )δ = 8.91 (br s, 1H), 8.58 (d, J = 5.2 Hz, 1H), 8.19 (d, J = 6.4 Hz, 1H), 7.29 (d, J = 4.8 Hz, 1H), 6.47-6.29 (m, 1H), 5.90 (dd, J = 1.2, 15.6 Hz, 1H), 2.69-2.56 (m, 1H), 1.74 (dd, J = 1.6, 6.4 Hz, 3H) , 1.15 (d, J = 6.8 Hz, 3H), 1.06 (d, J = 6.8 Hz, 3H).
步骤6：化合物001-8的合成Step 6: Synthesis of compound 001-8
氮气保护下，向预先干燥好的80mL长管中加入化合物001-7(1.00g,2.67mmol,1.00eq)和N,N'-二异丙基乙胺(1.72g,13.34mmol,2.32mL,5.00eq)。加完后，控温25℃，向体系中缓慢加入三氯氧磷(2.05g,13.34mmol,1.24mL,5.00eq)。加完后，体系在40℃搅拌2小时，得到化合物001-8。反应体系可直接用于下一步。Under the protection of nitrogen, add compound 001-7 (1.00g, 2.67mmol, 1.00eq) and N,N'-diisopropylethylamine (1.72g, 13.34mmol, 2.32mL, 5.00eq). After the addition, the temperature was controlled at 25°C, and phosphorus oxychloride (2.05g, 13.34mmol, 1.24mL, 5.00eq) was slowly added to the system. After the addition, the system was stirred at 40°C for 2 hours to obtain compound 001-8. The reaction system can be used directly in the next step.
LCMS:MS(ESI)m/z:393.0[M+1] +。 LCMS: MS (ESI) m/z: 393.0 [M+1] + .
步骤7：化合物001-10的合成Step 7: Synthesis of compound 001-10
氮气保护下，向含有化合物001-8(1.05g,2.67mmol,1.00eq)的80mL长管中加入四氢呋喃(40mL)，冷却到0℃，向体系中加入N,N'-二异丙基乙胺(5.18g,40.05mmol,6.98mL,15eq)。加完后，控温0℃，向体系中缓慢加入化合物001-9(802.14mg,4.01mmol,1.5eq)的四氢呋喃(20mL)溶液。加完后，体系在25℃搅拌1小时。将反应体系缓慢倒入10mL冰水中，加入15mL乙酸乙酯稀释。体系静置分液。分液后收集有机相，水相用乙酸乙酯萃取(10mL*1)。合并有机相用无水硫酸钠干燥，减压浓缩得到残余物。粗产品通过快速柱层析分离(石油醚:乙酸乙酯＝1:1至1:4)，纯化得到化合物001-10。Under the protection of nitrogen, tetrahydrofuran (40mL) was added to the 80mL long tube containing compound 001-8 (1.05g, 2.67mmol, 1.00eq), cooled to 0℃, and N,N'-diisopropylethyl was added to the system Amine (5.18g, 40.05mmol, 6.98mL, 15eq). After the addition, the temperature was controlled to 0°C, and a solution of compound 001-9 (802.14 mg, 4.01 mmol, 1.5 eq) in tetrahydrofuran (20 mL) was slowly added to the system. After the addition, the system was stirred at 25°C for 1 hour. The reaction system was slowly poured into 10 mL of ice water, and 15 mL of ethyl acetate was added to dilute. The system is allowed to stand for liquid separation. After liquid separation, the organic phase was collected, and the aqueous phase was extracted with ethyl acetate (10 mL*1). The combined organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The crude product was separated by flash column chromatography (petroleum ether: ethyl acetate=1:1 to 1:4) and purified to obtain compound 001-10.
LCMS:MS(ESI)m/z:557.2[M+1] +。 LCMS: MS (ESI) m/z: 557.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.53(d,J＝5.2Hz,1H),7.72(d,J＝7.6Hz,1H),7.28(dd,J＝2.8,5.2Hz,1H),6.42-6.27(m,1H),5.93-5.79(m,1H),4.90-4.55(m,1H),4.36-4.07(m,2H),4.01-3.77(m,1H),3.70-3.44(m,1H),3.36-2.90(m,2H),2.61-2.42(m,1H),1.68(td,J＝1.6,6.8Hz,4H),1.46-1.38(m,12H),1.14(dd,J＝4.4,6.8Hz,3H),1.03(t,J＝7.2Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.53 (d, J = 5.2 Hz, 1H), 7.72 (d, J = 7.6 Hz, 1H), 7.28 (dd, J = 2.8, 5.2 Hz, 1H), 6.42 -6.27(m,1H),5.93-5.79(m,1H),4.90-4.55(m,1H),4.36-4.07(m,2H),4.01-3.77(m,1H),3.70-3.44(m, 1H),3.36-2.90(m,2H),2.61-2.42(m,1H),1.68(td,J=1.6,6.8Hz,4H),1.46-1.38(m,12H),1.14(dd,J= 4.4, 6.8 Hz, 3H), 1.03 (t, J=7.2 Hz, 3H).
步骤8：化合物001-12的合成Step 8: Synthesis of compound 001-12
在预先干燥过的100mL单口瓶中加入化合物001-10(927mg,1.66mmol,1.00eq)，化合物001-11(272.44mg,1.75mmol,1.05eq)，磷酸钾(706.48mg,3.33mmol,2.0eq)，四氢呋喃(8mL)和H 2O(2mL)。加完后，体系氮气置换3次，向体系中加1,1'-双(二叔丁基膦)二茂铁二氯合钯(II)(108.46mg,166.41μmol,0.1eq)。加完后，体系氮气置换3次。将体系加热到80℃，反应在80℃搅拌12小时。体系直接浓缩。粗产品通过快速柱层析分离(石油醚:乙酸乙酯:甲醇＝2:1:0.5至4:1:0.5)纯化得到化合物001-12。 Add compound 001-10 (927mg, 1.66mmol, 1.00eq), compound 001-11 (272.44mg, 1.75mmol, 1.05eq), potassium phosphate (706.48mg, 3.33mmol, 2.0eq) into a pre-dried 100mL single-mouth flask ), tetrahydrofuran (8 mL) and H 2 O (2 mL). After the addition, the system was replaced with nitrogen for 3 times, and 1,1'-bis(di-tert-butylphosphine)ferrocene dichloropalladium(II) (108.46 mg, 166.41 μmol, 0.1 eq) was added to the system. After the addition, the system was replaced with nitrogen 3 times. The system was heated to 80°C, and the reaction was stirred at 80°C for 12 hours. The system is directly concentrated. The crude product was separated and purified by flash column chromatography (petroleum ether: ethyl acetate: methanol = 2:1:0.5 to 4:1:0.5) to obtain compound 001-12.
LCMS:MS(ESI)m/z:633.4[M+1] +。 LCMS: MS (ESI) m/z: 633.4 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝9.12(d,J＝6.0Hz,1H),8.57(d,J＝5.2Hz,1H),7.84(dd,J＝7.2,9.2 Hz,1H),7.35(dd,J＝1.2,5.2Hz,1H),7.24-7.16(m,1H),6.65-6.54(m,2H),6.47-6.34(m,1H),5.88(t,J＝16.4Hz,1H),5.12-4.67(m,1H),4.55-4.12(m,2H),4.01-3.80(m,1H),3.77-3.43(m,1H),3.36-2.94(m,2H),2.82-2.57(m,1H),1.71-1.63(m,3H),1.48-1.40(m,12H),1.24-1.13(m,6H)。 1 H NMR (400MHz, CDCl 3 )δ = 9.12 (d, J = 6.0 Hz, 1H), 8.57 (d, J = 5.2 Hz, 1H), 7.84 (dd, J = 7.2, 9.2 Hz, 1H), 7.35 (dd,J=1.2,5.2Hz,1H),7.24-7.16(m,1H),6.65-6.54(m,2H),6.47-6.34(m,1H),5.88(t,J=16.4Hz,1H ),5.12-4.67(m,1H),4.55-4.12(m,2H),4.01-3.80(m,1H),3.77-3.43(m,1H),3.36-2.94(m,2H),2.82-2.57 (m, 1H), 1.71-1.63 (m, 3H), 1.48-1.40 (m, 12H), 1.24-1.13 (m, 6H).
步骤9：化合物001-13的合成Step 9: Synthesis of compound 001-13
在预先干燥过的40mL单口瓶中加入化合物001-12(0.35g,553.19μmol,1eq)，烯丙基溴(80.31mg,663.82μmol,1.2eq)，碳酸钾(114.68mg,829.78μmol,1.5eq)，碘化钾(119.38mg,719.14μmol,1.3eq)和乙腈(20mL)。加完后，反应在25℃搅拌12小时。向体系中加入10mL水和10mL乙酸乙酯。体系静置分液。水相用乙酸乙酯萃取(10mL)。合并有机相，用饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤。滤液浓缩得到化合物001-13。粗品无需纯化直接用于下一步。Add compound 001-12 (0.35g, 553.19μmol, 1eq), allyl bromide (80.31mg, 663.82μmol, 1.2eq), potassium carbonate (114.68mg, 829.78μmol, 1.5eq) into a pre-dried 40mL single-mouth bottle ), potassium iodide (119.38 mg, 719.14 μmol, 1.3 eq) and acetonitrile (20 mL). After the addition, the reaction was stirred at 25°C for 12 hours. Add 10 mL of water and 10 mL of ethyl acetate to the system. The system is allowed to stand for liquid separation. The aqueous phase was extracted with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to obtain compound 001-13. The crude product is used directly in the next step without purification.
LCMS:MS(ESI)m/z:673.2[M+1] +。 LCMS: MS (ESI) m/z: 673.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.42(d,J＝4.8Hz,1H),7.73(d,J＝8.0Hz,1H),7.25-7.14(m,2H),6.69-6.52(m,2H),6.34-6.15(m,1H),5.90(t,J＝16.0Hz,1H),5.78-5.54(m,1H),5.14-4.95(m,2H),4.93-4.68(m,1H),4.51-4.14(m,3H),4.13-3.78(m,2H),3.74-3.42(m,1H),3.36-2.90(m,2H),2.75-2.47(m,1H),1.73-1.62(m,3H),1.45(s,12H),1.17-1.09(m,3H),1.02-0.85(m,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.42 (d, J = 4.8 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.25-7.14 (m, 2H), 6.69-6.52 (m, 2H), 6.34-6.15 (m, 1H), 5.90 (t, J=16.0Hz, 1H), 5.78-5.54 (m, 1H), 5.14-4.95 (m, 2H), 4.93-4.68 (m, 1H) ,4.51-4.14(m,3H),4.13-3.78(m,2H),3.74-3.42(m,1H),3.36-2.90(m,2H),2.75-2.47(m,1H),1.73-1.62( m, 3H), 1.45 (s, 12H), 1.17-1.09 (m, 3H), 1.02-0.85 (m, 3H).
步骤10：化合物001-14的合成Step 10: Synthesis of compound 001-14
在预先干燥过的100mL单口瓶中加入化合物001-13(377mg,560.38μmol,1eq),(1,3-双(2,4,6-三甲基苯基)-2-咪唑烷亚基)二氯(邻异丙氧基苯亚甲基)合钌(134.64mg,158.59μmol,0.283eq)和二氯甲烷(45mL)。加完后，反应在45℃搅拌1.0小时。反应体系直接浓缩。粗产品通过快速柱层析分离(石油醚:乙酸乙酯:甲醇＝1:1:0.02至6:1:0.02)，纯化得到化合物001-14。Add compound 001-13 (377mg, 560.38μmol, 1eq), (1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene) into a pre-dried 100mL single-mouth bottle Dichloro(o-isopropoxybenzylidene)ruthenium (134.64mg, 158.59μmol, 0.283eq) and dichloromethane (45mL). After the addition, the reaction was stirred at 45°C for 1.0 hour. The reaction system is directly concentrated. The crude product was separated by flash column chromatography (petroleum ether: ethyl acetate: methanol=1:1:0.02 to 6:1:0.02) and purified to obtain compound 001-14.
LCMS:MS(ESI)m/z:631.3[M+1] +。 LCMS: MS (ESI) m/z: 631.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.53(d,J＝4.8Hz,1H),7.76(dd,J＝8.4,12.8Hz,1H),7.39-7.26(m,1H),7.00-6.82(m,2H),6.51(t,J＝10.8Hz,1H),6.04-5.68(m,1H),4.81-4.56(m,1H),4.40-4.13(m,2H),4.06-3.86(m,1H),3.84-3.45(m,1H),3.42-2.96(m,2H),2.94-2.68(m,1H),2.37-2.14(m,1H),1.96-1.66(m,2H),1.68-1.54(m,3H),1.50(s,9H),1.31-1.17(m,6H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.53 (d, J = 4.8 Hz, 1H), 7.76 (dd, J = 8.4, 12.8 Hz, 1H), 7.39-7.26 (m, 1H), 7.00-6.82 ( m, 2H), 6.51 (t, J = 10.8 Hz, 1H), 6.04-5.68 (m, 1H), 4.81-4.56 (m, 1H), 4.40-4.13 (m, 2H), 4.06-3.86 (m, 1H), 3.84-3.45 (m, 1H), 3.42-2.96 (m, 2H), 2.94-2.68 (m, 1H), 2.37-2.14 (m, 1H), 1.96-1.66 (m, 2H), 1.68- 1.54 (m, 3H), 1.50 (s, 9H), 1.31-1.17 (m, 6H).
步骤11：化合物001-15的三氟乙酸盐的合成Step 11: Synthesis of trifluoroacetate salt of compound 001-15
氮气保护下，向预先干燥好的100mL三口瓶中加入化合物001-14(0.15g,237.84μmol,1eq)和二氯甲烷(9mL)。加完后，用注射器向体系中滴加三氟乙酸(271.19mg,2.38mmol,176.10μL,10eq)。加完后，体系在25℃，搅拌3小时。将体系减压浓缩得到化合物001-15的三氟乙酸盐。粗品无需纯化可直接用于下一步反应。Under the protection of nitrogen, compound 001-14 (0.15 g, 237.84 μmol, 1 eq) and dichloromethane (9 mL) were added to a pre-dried 100 mL three-necked flask. After the addition, trifluoroacetic acid (271.19mg, 2.38mmol, 176.10μL, 10eq) was added dropwise to the system with a syringe. After the addition, the system was stirred at 25°C for 3 hours. The system was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 001-15. The crude product can be used directly in the next reaction without purification.
LCMS:MS(ESI)m/z:531.3[M+1] +。 LCMS: MS (ESI) m/z: 531.3 [M+1] + .
步骤12：化合物001和002的合成Step 12: Synthesis of compounds 001 and 002
在预先干燥过的100mL单口瓶中加入化合物001-15(0.26g,229.10μmol,1.0eq,5.3TFA)，N,N'-二异丙基乙胺(296.09mg,2.29mmol,399.04μL,10eq)和二氯甲烷(10mL)。加完后，体系冷却到-60℃。用注射器向体系中滴加丙烯酰氯(20.74mg,229.10μmol,1.0eq)。加完后反应在-60℃搅拌2分钟。 检测。将体系缓慢加水(10mL)，静置分液。分液后收集有机相，水相用二氯甲烷萃取(10mL*1)。合并有机相，用无水硫酸钠干燥，减压浓缩得到粗品。粗品经制备分离(分离方法：柱子:Phenomenex Gemini-NX 80*40mm*3μm；流动相:[水(10mM碳酸氢铵)-乙腈]；B(乙腈)％:25％-55％,8min)纯化。馏分冻干。产品经SFC拆分(分离方法：柱子:DAICEL CHIRALPAK AD(250mm*30mm,10μm)；流动相:[0.1％氨水-乙醇]；B(乙醇)％:45％-45％,12min)得到化合物001和002。Add compound 001-15 (0.26g, 229.10μmol, 1.0eq, 5.3TFA), N,N'-diisopropylethylamine (296.09mg, 2.29mmol, 399.04μL, 10eq ) And dichloromethane (10 mL). After the addition, the system was cooled to -60°C. Add acryloyl chloride (20.74mg, 229.10μmol, 1.0eq) into the system dropwise with a syringe. After the addition, the reaction was stirred at -60°C for 2 minutes. Detection. Slowly add water (10 mL) to the system and let stand for liquid separation. After liquid separation, the organic phase was collected, and the aqueous phase was extracted with dichloromethane (10 mL*1). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was prepared and separated (separation method: column: Phenomenex Gemini-NX 80*40mm*3μm; mobile phase: [water (10mM ammonium bicarbonate)-acetonitrile]; B (acetonitrile)%: 25%-55%, 8min) purification . The fraction is lyophilized. The product was resolved by SFC (Separation method: column: DAICEL CHIRALPAK AD (250mm*30mm, 10μm); mobile phase: [0.1% ammonia-ethanol]; B (ethanol)%: 45%-45%, 12min) to obtain compound 001 And 002.
化合物001：手性柱出峰位置：1.16minCompound 001: Peak position of chiral column: 1.16min
LCMS:MS(ESI)m/z:585.2[M+1] +。 LCMS: MS (ESI) m/z: 585.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(d,J＝4.8Hz,1H),7.70(d,J＝8.4Hz,1H),7.31(dd,J＝8.4,15.2Hz,1H),6.90(d,J＝8.4Hz,1H),6.86(t,J＝8.4Hz,1H),6.80(d,J＝4.8Hz,1H),6.66-6.48(m,1H),6.44(d,J＝10.4Hz,1H),6.35(d,J＝16.8Hz,1H),5.75(d,J＝10.4Hz,1H),5.73-5.63(m,1H),4.90-4.62(m,2H),4.58(dd,J＝2.4,9.6Hz,1H),4.42(d,J＝12.0Hz,1H),4.15(t,J＝10.4Hz,1H),4.03-3.76(m,1H),3.71-3.32(m,2H),3.21-3.04(m,1H),2.87-2.73(m,1H),1.61-1.53(m,3H),1.23(d,J＝6.8Hz,3H),0.92(d,J＝6.4Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.49 (d, J = 4.8 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 7.31 (dd, J = 8.4, 15.2 Hz, 1H), 6.90 (d,J=8.4Hz,1H), 6.86(t,J=8.4Hz,1H), 6.80(d,J=4.8Hz,1H),6.66-6.48(m,1H),6.44(d,J= 10.4Hz, 1H), 6.35 (d, J = 16.8 Hz, 1H), 5.75 (d, J = 10.4 Hz, 1H), 5.73-5.63 (m, 1H), 4.90-4.62 (m, 2H), 4.58 ( dd, J = 2.4, 9.6 Hz, 1H), 4.42 (d, J = 12.0 Hz, 1H), 4.15 (t, J = 10.4 Hz, 1H), 4.03-3.76 (m, 1H), 3.71-3.32 (m ,2H),3.21-3.04(m,1H),2.87-2.73(m,1H),1.61-1.53(m,3H),1.23(d,J=6.8Hz,3H),0.92(d,J=6.4 Hz, 3H).
化合物002：手性柱出峰位置：1.37minCompound 002: Peak position of chiral column: 1.37min
LCMS:MS(ESI)m/z:585.2[M+1] +。 LCMS: MS (ESI) m/z: 585.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(d,J＝4.8Hz,1H),7.73(d,J＝8.0Hz,1H),7.31(dd,J＝8.0,15.2Hz,1H),6.91(d,J＝8.0Hz,1H),6.86(t,J＝8.4Hz,1H),6.81(d,J＝4.8Hz,1H),6.65-6.42(m,2H),6.40-6.29(m,1H),5.79-5.65(m,2H),5.31-4.41(m,3H),4.30-3.96(m,2H),3.91-3.16(m,3H),2.98-2.72(m,1H),1.38-1.31(m,3H),1.23(d,J＝6.8Hz,3H),0.98-0.84(m,3H)。 1 H NMR (400MHz, CDCl 3 )δ = 8.49 (d, J = 4.8 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.31 (dd, J = 8.0, 15.2 Hz, 1H), 6.91 (d,J=8.0Hz,1H), 6.86(t,J=8.4Hz,1H), 6.81(d,J=4.8Hz,1H),6.65-6.42(m,2H),6.40-6.29(m, 1H), 5.79-5.65 (m, 2H), 5.31-4.41 (m, 3H), 4.30-3.96 (m, 2H), 3.91-3.16 (m, 3H), 2.98-2.72 (m, 1H), 1.38- 1.31 (m, 3H), 1.23 (d, J=6.8 Hz, 3H), 0.98-0.84 (m, 3H).
实施例2Example 2
Figure PCTCN2021073149-appb-000046
Figure PCTCN2021073149-appb-000046
合成路线：synthetic route:
Figure PCTCN2021073149-appb-000047
Figure PCTCN2021073149-appb-000047
步骤1：化合物003-1的合成Step 1: Synthesis of compound 003-1
用氩气置换过的35mL氢化瓶加入钯碳(0.2g,10％纯度)。加完后，将甲醇(20mL)和001-14(0.24g,380.54μmol,1eq)加入到体系中。加料完毕，将体系用氢气置换3次。体系氢气氛围中在25℃(20psi)搅拌12小时。体系滤过一层硅藻土。浓缩得到化合物003-1。粗品无需纯化可直接用于下一步反应。A 35 mL hydrogenation flask replaced with argon was added with palladium on carbon (0.2 g, 10% purity). After the addition, methanol (20mL) and 001-14 (0.24g, 380.54μmol, 1eq) were added to the system. After the feeding was completed, the system was replaced with hydrogen three times. The system was stirred for 12 hours at 25°C (20 psi) in a hydrogen atmosphere. The system was filtered through a layer of diatomaceous earth. Concentrate to obtain compound 003-1. The crude product can be used directly in the next reaction without purification.
LCMS:MS(ESI)m/z:633.3[M+1] +。 LCMS: MS (ESI) m/z: 633.3 [M+1] + .
步骤2：化合物003-2的三氟乙酸盐的合成Step 2: Synthesis of the trifluoroacetate salt of compound 003-2
氮气保护下，向预先干燥好的100mL单口瓶中加入化合物003-1(0.24g,379.33μmol,1eq)和二氯甲烷(5mL)。加完后，用注射器向体系中滴加三氟乙酸(432.51mg,3.79mmol,280.85μL,10eq)。加完后，体系在25℃，搅拌1.0小时。将体系减压浓缩得到化合物003-2的三氟乙酸盐。粗品无需纯化可直接用于下一步反应。Under the protection of nitrogen, compound 003-1 (0.24 g, 379.33 μmol, 1 eq) and dichloromethane (5 mL) were added to a pre-dried 100 mL single-neck flask. After the addition, trifluoroacetic acid (432.51mg, 3.79mmol, 280.85μL, 10eq) was added dropwise to the system with a syringe. After the addition, the system was stirred at 25°C for 1.0 hour. The system was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 003-2. The crude product can be used directly in the next reaction without purification.
LCMS:MS(ESI)m/z:533.3[M+1] +。 LCMS: MS (ESI) m/z: 533.3 [M+1] + .
步骤3：化合物003和004的合成Step 3: Synthesis of compounds 003 and 004
在预先干燥过的100mL单口瓶中加入化合物003-2(0.420g,376.60μmol,1.0eq,折算5.11个三氟乙酸盐)，N,N'-二异丙基乙胺(486.72mg,3.77mmol,655.95μL,10eq)和二氯甲烷(10mL)。加完后，体系冷却到-60℃。用注射器向体系中滴加丙烯酰氯(34.09mg,376.60μmol,1.0eq)。加完后反应在-60℃搅拌2分钟。将体系缓慢加水(10mL)，静置分液。分液后收集有机相，水相用二氯甲烷萃取(10mL)。合并有机相，用无水硫酸钠干燥，减压浓缩得到粗品。粗品经制备分离(分离方法：柱子:Welch Xtimate C18 150*25mm*5μm；流动相:[水(0.04％盐酸)-乙腈]；B(乙腈)％:20％-35％,8min)纯化，馏分冻干。 产品用SFC分离(分离方法：柱子:REGIS(s,s)WHELK-O1(250mm*50mm,10μm)；流动相:[0.1％氨水-乙醇]；B(乙醇)％:55％-55％,6min)，馏分浓缩，加去离子水冻干得到003(即计算例1的化合物WX001)和004。Add compound 003-2 (0.420g, 376.60μmol, 1.0eq, equivalent to 5.11 trifluoroacetate), N,N'-diisopropylethylamine (486.72mg, 3.77 mmol, 655.95 μL, 10 eq) and dichloromethane (10 mL). After the addition, the system was cooled to -60°C. Use a syringe to add acryloyl chloride (34.09 mg, 376.60 μmol, 1.0 eq) to the system dropwise. After the addition, the reaction was stirred at -60°C for 2 minutes. Slowly add water (10 mL) to the system and let stand for liquid separation. After liquid separation, the organic phase was collected, and the aqueous phase was extracted with dichloromethane (10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product is prepared and separated (separation method: column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.04% hydrochloric acid)-acetonitrile]; B (acetonitrile)%: 20%-35%, 8min) purification, fractions Freeze-dried. The product is separated by SFC (Separation method: column: REGIS(s, s) WHELK-O1 (250mm*50mm, 10μm); mobile phase: [0.1% ammonia-ethanol]; B (ethanol)%: 55%-55%, 6min), the fractions were concentrated, and deionized water was added to freeze-dry to obtain 003 (ie, compound WX001 of calculation example 1) and 004.
化合物003：手性柱出峰位置：3.157minCompound 003: Peak position of chiral column: 3.157min
LCMS:MS(ESI)m/z:587.2[M+1] +。 LCMS: MS (ESI) m/z: 587.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(d,J＝5.2Hz,1H),7.72(d,J＝8.4Hz,1H),7.29(td,J＝6.8,8.0Hz,1H),7.04(d,J＝5.2Hz,1H),6.83(d,J＝8.4Hz,1H),6.76(t,J＝8.8Hz,1H),6.65-6.39(m,1H),6.35(dd,J＝1.6,16.8Hz,1H),5.75(d,J＝1.2,10.0Hz,1H),4.88-4.36(m,3H),4.33-4.26(m,1H),4.01-3.74(m,1H),3.72-3.30(m,3H),3.24-3.03(m,1H),2.91-2.78(m,1H),2.55-2.45(m,1H),2.43-2.31(m,1H),2.25-2.01(m,2H),1.31-1.10(m,6H),0.97(d,J＝6.8Hz,3H)。 1 H NMR (400MHz, CDCl 3 )δ = 8.49 (d, J = 5.2 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.29 (td, J = 6.8, 8.0 Hz, 1H), 7.04 (d,J=5.2Hz,1H), 6.83(d,J=8.4Hz,1H), 6.76(t,J=8.8Hz,1H),6.65-6.39(m,1H), 6.35(dd,J= 1.6, 16.8 Hz, 1H), 5.75 (d, J = 1.2, 10.0 Hz, 1H), 4.88-4.36 (m, 3H), 4.33-4.26 (m, 1H), 4.01-3.74 (m, 1H), 3.72 -3.30(m,3H),3.24-3.03(m,1H),2.91-2.78(m,1H),2.55-2.45(m,1H),2.43-2.31(m,1H),2.25-2.01(m, 2H), 1.31-1.10 (m, 6H), 0.97 (d, J=6.8 Hz, 3H).
化合物004：手性柱出峰位置：3.836minCompound 004: Peak position of chiral column: 3.836min
LCMS:MS(ESI)m/z:587.2[M+1] +. LCMS:MS(ESI)m/z:587.2[M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(d,J＝5.2Hz,1H),7.76(br d,J＝8.4Hz,1H),7.34-7.24(m,1H),7.04(d,J＝4.8Hz,1H),6.83(d,J＝8.0Hz,1H),6.75(t,J＝8.8Hz,1H),6.63-6.45(m,1H),6.35(dd,J＝1.6,16.8Hz,1H),5.75(dd,J＝1.6,10.4Hz,1H),5.42-4.95(m,1H),4.80-4.45(m,1H),4.34-4.25(m,1H),4.17-4.07(m,1H),3.94-3.79(m,1H),3.78-3.64(m,1H),3.63-3.52(m,1H),3.50-3.20(m,1H),2.92-2.82(m,1H),2.56-2.45(m,1H),2.44-2.30(m,1H),2.25-2.00(m,2H),1.30(d,J＝6.4Hz,3H),1.24(d,J＝6.4Hz,3H),1.18(s,1H),0.96(br d,J＝5.2Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.49 (d, J = 5.2 Hz, 1H), 7.76 (br d, J = 8.4 Hz, 1H), 7.34-7.24 (m, 1H), 7.04 (d, J ＝4.8Hz,1H), 6.83(d,J＝8.0Hz,1H), 6.75(t,J＝8.8Hz,1H),6.63-6.45(m,1H),6.35(dd,J＝1.6,16.8Hz ,1H), 5.75(dd,J=1.6,10.4Hz,1H),5.42-4.95(m,1H),4.80-4.45(m,1H),4.34-4.25(m,1H),4.17-4.07(m ,1H),3.94-3.79(m,1H),3.78-3.64(m,1H),3.63-3.52(m,1H),3.50-3.20(m,1H),2.92-2.82(m,1H),2.56 -2.45(m,1H),2.44-2.30(m,1H),2.25-2.00(m,2H),1.30(d,J=6.4Hz,3H), 1.24(d,J=6.4Hz,3H), 1.18 (s, 1H), 0.96 (br d, J=5.2 Hz, 3H).
实施例3Example 3
Figure PCTCN2021073149-appb-000048
Figure PCTCN2021073149-appb-000048
合成路线：synthetic route:
Figure PCTCN2021073149-appb-000049
Figure PCTCN2021073149-appb-000049
步骤1：化合物005-1的合成Step 1: Synthesis of compound 005-1
氮气保护下，将化合物001-12(0.37g,584.80μmol,1eq)和化合物4-溴-1-丁烯(789.49mg,5.85mmol,593.60μL,10eq)溶于乙腈(20mL)，加入碳酸钾(181.86mg,1.32mmol,2.25eq)和碘化钾(189.30mg,1.14mmol,1.95eq)，80℃反应5小时。补加化合物4-溴-1-丁烯(394.74mg,2.92mmol,296.80μL,5eq)，80℃反应5小时。向反应体系中加入水(20mL)，水相使用乙酸乙酯(20mL*2)萃取，分液。合并有机相，使用饱和食盐水(30mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过快速柱层析分离(流动相：甲醇/二氯甲烷＝0.2％～0.8％)分离，纯化得到化合物005-1。Under nitrogen protection, compound 001-12 (0.37g, 584.80μmol, 1eq) and compound 4-bromo-1-butene (789.49mg, 5.85mmol, 593.60μL, 10eq) were dissolved in acetonitrile (20mL), and potassium carbonate was added (181.86mg, 1.32mmol, 2.25eq) and potassium iodide (189.30mg, 1.14mmol, 1.95eq) were reacted at 80°C for 5 hours. Compound 4-bromo-1-butene (394.74mg, 2.92mmol, 296.80μL, 5eq) was added and reacted at 80°C for 5 hours. Water (20 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (20 mL*2) and separated. The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by flash column chromatography (mobile phase: methanol/dichloromethane = 0.2% to 0.8%) and purified to obtain compound 005-1.
LCMS:MS(ESI)m/z:687.3[M+1] +。 LCMS: MS (ESI) m/z: 687.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.51(d,J＝5.2Hz,1H),7.79(t,J＝8.8Hz,1H),7.34-7.28(m,2H),6.70(d,J＝8.4Hz,2H),6.40-6.31(m,1H),5.98(t,J＝16.8Hz,1H),5.65(s,1H),4.99-4.96(m,3H),4.47-4.24(m,2H),4.14-4.06(m,1H),3.98-3.89(m,2H),3.71-3.60(m,1H),3.35-3.15(m,2H),2.73-2.69(m,1H),2.32-2.27(m,2H),1.76(t,J＝6.8Hz,3H),1.60(s,6H),1.53(s,9H),1.05-0.97(m,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.51 (d, J = 5.2 Hz, 1H), 7.79 (t, J = 8.8 Hz, 1H), 7.34-7.28 (m, 2H), 6.70 (d, J = 8.4Hz, 2H), 6.40-6.31 (m, 1H), 5.98 (t, J = 16.8 Hz, 1H), 5.65 (s, 1H), 4.99-4.96 (m, 3H), 4.47-4.24 (m, 2H) ),4.14-4.06(m,1H),3.98-3.89(m,2H),3.71-3.60(m,1H),3.35-3.15(m,2H),2.73-2.69(m,1H),2.32-2.27 (m, 2H), 1.76 (t, J=6.8 Hz, 3H), 1.60 (s, 6H), 1.53 (s, 9H), 1.05-0.97 (m, 3H).
步骤2：化合物005-2的合成Step 2: Synthesis of compound 005-2
氮气保护，将化合物005-1(0.29g,422.25μmol,1eq)溶于二氯甲烷(45mL)，加入(1,3-双(2,4,6-三甲基苯基)-2-咪唑烷亚基)二氯(邻异丙氧基苯亚甲基)合钌(71.70mg,84.45μmol,0.2eq)，45℃反应1.5小时。因未反应完全，45℃反应1.5小时。将反应体系直接减压浓缩得到粗产品。粗产品通过快速柱层析分离(流动相：甲醇/二氯甲烷＝0.2％～1.5％)分离，纯化得到化合物005-2。Under nitrogen protection, compound 005-1 (0.29g, 422.25μmol, 1eq) was dissolved in dichloromethane (45mL), and (1,3-bis(2,4,6-trimethylphenyl)-2-imidazole Alkylene) dichloro(o-isopropoxybenzylidene)ruthenium (71.70mg, 84.45μmol, 0.2eq), reacted at 45°C for 1.5 hours. The reaction was not completed at 45°C for 1.5 hours. The reaction system was directly concentrated under reduced pressure to obtain a crude product. The crude product was separated by flash column chromatography (mobile phase: methanol/dichloromethane = 0.2% to 1.5%) and purified to obtain compound 005-2.
LCMS:MS(ESI)m/z:645.3[M+1] +。 LCMS: MS (ESI) m/z: 645.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.54(d,J＝5.2Hz,1H),7.77(dd,J＝19.6,8.0Hz,1H),7.38-7.32(m,2H),6.74(t,J＝8.8Hz,1H),6.66(d,J＝8.4Hz,1H),6.12-6.05(m,1H),6.02-5.97(m,1H),4.70-4.64(m,1H),4.34-4.27(m,1H),4.19(d,J＝8.8Hz,1H),4.12-4.08(m,1H),4.01-3.96(m,1H),3.91(t,J＝9.2Hz,1H),3.85-3.74(m,1H),2.95-2.91(m,1H),2.49-2.35(m,2H),1.91-1.75(m,2H),1.59(s,6H),1.52(s,9H),1.34(d,J＝6.4Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.54 (d, J = 5.2 Hz, 1H), 7.77 (dd, J = 19.6, 8.0 Hz, 1H), 7.38-7.32 (m, 2H), 6.74 (t, J=8.8Hz,1H),6.66(d,J=8.4Hz,1H),6.12-6.05(m,1H),6.02-5.97(m,1H),4.70-4.64(m,1H),4.34-4.27 (m,1H), 4.19(d,J=8.8Hz,1H),4.12-4.08(m,1H),4.01-3.96(m,1H),3.91(t,J=9.2Hz,1H),3.85- 3.74(m, 1H), 2.95-2.91(m, 1H), 2.49-2.35(m, 2H), 1.91-1.75(m, 2H), 1.59(s, 6H), 1.52(s, 9H), 1.34( d, J=6.4 Hz, 3H).
步骤3：化合物005-3的合成Step 3: Synthesis of compound 005-3
将化合物005-2(280.00mg,434.30μmol,1eq)溶于甲醇(30mL)，加入钯碳(0.6g,10％纯度)，氢气置换三次，然后氢气氛围下(压力：50Psi)，50℃反应15小时。LCMS检测未反应完全，50℃继续反应5小时。将反应体系直接过滤，滤液减压浓缩得到化合物005-3。粗品无需纯化可直接用于下一步反应。Compound 005-2 (280.00mg, 434.30μmol, 1eq) was dissolved in methanol (30mL), palladium on carbon (0.6g, 10% purity) was added, hydrogen was replaced three times, and then reacted at 50℃ under hydrogen atmosphere (pressure: 50Psi) 15 hours. LCMS detected that the reaction was not complete, and the reaction was continued at 50°C for 5 hours. The reaction system was directly filtered, and the filtrate was concentrated under reduced pressure to obtain compound 005-3. The crude product can be used directly in the next reaction without purification.
LCMS:MS(ESI)m/z:647.2[M+1] +. LCMS:MS(ESI)m/z:647.2[M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.48(d,J＝4.8Hz,1H),7.85-7.82(m,1H),7.35(q,J＝8.0Hz,1H),7.02(d,J＝5.2Hz,1H),6.84(d,J＝8.4Hz,1H),6.80(t,J＝8.8Hz,1H),5.01-4.84(m,1H),4.31-4.26(m,1H),3.94-3.89(m,3H),3.74-3.61(m,1H),3.39-3.13(m,3H),2.97-2.87(m,1H),2.69-2.57(m,1H),2.49-2.42(m,1H),1.58(s,6H),1.53(s,9H),1.20(dd,J＝6.8,3.6Hz,4H),0.96(t,J＝6.8Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.48 (d, J = 4.8 Hz, 1H), 7.85-7.82 (m, 1H), 7.35 (q, J = 8.0 Hz, 1H), 7.02 (d, J = 5.2Hz, 1H), 6.84 (d, J = 8.4 Hz, 1H), 6.80 (t, J = 8.8 Hz, 1H), 5.01-4.84 (m, 1H), 4.31-4.26 (m, 1H), 3.94 3.89 (m, 3H), 3.74-3.61 (m, 1H), 3.39-3.13 (m, 3H), 2.97-2.87 (m, 1H), 2.69-2.57 (m, 1H), 2.49-2.42 (m, 1H) ), 1.58 (s, 6H), 1.53 (s, 9H), 1.20 (dd, J = 6.8, 3.6 Hz, 4H), 0.96 (t, J = 6.8 Hz, 3H).
步骤4：化合物005-4的三氟乙酸盐的合成Step 4: Synthesis of the trifluoroacetate salt of compound 005-4
氮气保护下，将化合物005-3(0.24g,371.10μmol,1eq)溶于二氯甲烷(24mL)，加入三氟乙酸(2.12g,18.56mmol,1.37mL,50eq)，20℃反应3小时。将反应体系直接减压浓缩，得到化合物005-4的三氟乙酸盐。该反应体系无纯化，直接用于下一步反应。Under nitrogen protection, compound 005-3 (0.24 g, 371.10 μmol, 1 eq) was dissolved in dichloromethane (24 mL), trifluoroacetic acid (2.12 g, 18.56 mmol, 1.37 mL, 50 eq) was added, and the reaction was carried out at 20° C. for 3 hours. The reaction system was directly concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 005-4. The reaction system was not purified and was directly used in the next reaction.
LCMS:MS(ESI)m/z:547.2[M+1] +. LCMS:MS(ESI)m/z:547.2[M+1] + .
步骤5：化合物005和006的合成Step 5: Synthesis of compounds 005 and 006
氮气保护下，将化合物005-4(0.45g,334.63μmol,1eq,折算7个三氟乙酸盐)溶于二氯甲烷(30mL)，加入N,N'-二异丙基乙胺(518.97mg,4.02mmol,699.42μL,12eq)，然后加入丙烯酰氯(45.43mg,501.94μmol,40.93μL,1.5eq)，-60℃反应0.5小时。向反应体系中加入饱和碳酸氢钠溶液(20mL)，分液。水相使用二氯甲烷(20mL*3)萃取，分液。合并有机相，使用饱和食盐水(30mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过HPLC(柱子:Phenomenex luna C18 80*40mm*3μm；流动相:[水(0.04％盐酸)-乙腈]；B(乙腈)％:18％-36％,7min)分离，然后进一步通过SFC(柱子:DAICEL CHIRALPAK IG(250mm*30mm,10μm)；流动相:[0.1％氨水-乙醇]；B(乙醇)％:60％-60％,3min)分离，纯化分别得到化合物005(即计算例1的化合物WX006)和化合物006。Under the protection of nitrogen, compound 005-4 (0.45g, 334.63μmol, 1eq, equivalent to 7 trifluoroacetate) was dissolved in dichloromethane (30mL), and N,N'-diisopropylethylamine (518.97) was added. mg, 4.02mmol, 699.42μL, 12eq), then add acryloyl chloride (45.43mg, 501.94μmol, 40.93μL, 1.5eq), and react at -60°C for 0.5 hours. Saturated sodium bicarbonate solution (20 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (20 mL*3) and separated. The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by HPLC (column: Phenomenex luna C18 80*40mm*3μm; mobile phase: [water (0.04% hydrochloric acid)-acetonitrile]; B (acetonitrile)%: 18%-36%, 7min), and then further passed by SFC (Column: DAICEL CHIRALPAK IG (250mm*30mm, 10μm); mobile phase: [0.1% ammonia-ethanol]; B (ethanol)%: 60%-60%, 3min) separation and purification to obtain compound 005 (ie calculation example) 1 compound WX006) and compound 006.
化合物005：手性柱出峰位置：1.839minCompound 005: Peak position of chiral column: 1.839min
LCMS:MS(ESI)m/z:601.3[M+1] +。 LCMS: MS (ESI) m/z: 601.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(d,J＝4.8Hz,1H),7.84(d,J＝8.4Hz,1H),7.35(q,J＝8.4Hz,1H),7.02(d,J＝5.2Hz,1H),6.84(d,J＝8.4Hz,1H),6.80(t,J＝8.8Hz,1H),6.69-6.56(m,1H),6.42(dd,J＝16.8,1.2Hz,1H),5.82(dd,J＝10.4,1.6Hz,1H),5.02-4.66(m,2H),4.54-4.41(m,1H),4.06-3.88(m, 3H),3.78-3.56(m,2H),3.27-3.11(m,1H),2.92-2.84(m,1H),2.68-2.58(m,1H),2.50-2.43(m,1H),1.53(d,J＝6.4Hz,3H),1.48-1.35(m,4H),1.20(d,J＝6.8Hz,3H),0.96(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.49 (d, J = 4.8 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.35 (q, J = 8.4 Hz, 1H), 7.02 (d ,J=5.2Hz,1H), 6.84(d,J=8.4Hz,1H), 6.80(t,J=8.8Hz,1H), 6.69-6.56(m,1H), 6.42(dd,J=16.8, 1.2Hz, 1H), 5.82 (dd, J = 10.4, 1.6 Hz, 1H), 5.02-4.66 (m, 2H), 4.54-4.41 (m, 1H), 4.06-3.88 (m, 3H), 3.78-3.56 (m,2H), 3.27-3.11(m,1H), 2.92-2.84(m,1H), 2.68-2.58(m,1H), 2.50-2.43(m,1H), 1.53(d,J=6.4Hz , 3H), 1.48-1.35 (m, 4H), 1.20 (d, J = 6.8 Hz, 3H), 0.96 (s, 3H).
化合物006：手性柱出峰位置：2.180minCompound 006: Peak position of chiral column: 2.180min
LCMS:MS(ESI)m/z:601.3[M+1] +。 LCMS: MS (ESI) m/z: 601.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.51(d,J＝5.2Hz,1H),7.86(d,J＝8.0Hz,1H),7.36(q,J＝6.8Hz,1H),7.07(d,J＝4.8Hz,1H),6.84(d,J＝8.4Hz,1H),6.80(t,J＝8.8Hz,1H),6.67-6.57(m,1H),6.42(dd,J＝16.4,0.8Hz,1H),5.83(dd,J＝10.4,1.6Hz,1H),4.91-4.77(m,1H),4.55-4.45(m,1H),4.32-4.07(m,1H),3.97-3.87(m,3H),3.76-3.62(m,2H),3.34-3.04(m,1H),2.92(d,J＝11.6Hz,1H),2.67-2.59(m,1H),2.52-2.45(m,1H),1.47(s,7H),1.22(d,J＝6.4Hz,3H),0.97(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.51 (d, J = 5.2 Hz, 1H), 7.86 (d, J = 8.0 Hz, 1H), 7.36 (q, J = 6.8 Hz, 1H), 7.07 (d ,J=4.8Hz,1H), 6.84(d,J=8.4Hz,1H), 6.80(t,J=8.8Hz,1H), 6.67-6.57(m,1H), 6.42(dd,J=16.4, 0.8Hz, 1H), 5.83 (dd, J = 10.4, 1.6 Hz, 1H), 4.91-4.77 (m, 1H), 4.55-4.45 (m, 1H), 4.32-4.07 (m, 1H), 3.97-3.87 (m,3H),3.76-3.62(m,2H),3.34-3.04(m,1H),2.92(d,J=11.6Hz,1H),2.67-2.59(m,1H),2.52-2.45(m , 1H), 1.47 (s, 7H), 1.22 (d, J = 6.4 Hz, 3H), 0.97 (s, 3H).
实施例4Example 4
Figure PCTCN2021073149-appb-000050
Figure PCTCN2021073149-appb-000050
合成路线：synthetic route:
Figure PCTCN2021073149-appb-000051
Figure PCTCN2021073149-appb-000051
步骤1：化合物007-1的三氟乙酸盐的合成Step 1: Synthesis of the trifluoroacetate salt of compound 007-1
氮气保护，将化合物005-2(160.00mg,248.17μmol,1eq)溶于二氯甲烷(16mL)，加入三氟乙酸(1.41g,12.41mmol,918.75μL,50eq)，20℃反应3小时。将反应体系直接减压浓缩得到粗产品化合物007-1的三氟乙酸盐，无需纯化可直接用于下一步反应。Under nitrogen protection, compound 005-2 (160.00 mg, 248.17 μmol, 1 eq) was dissolved in dichloromethane (16 mL), trifluoroacetic acid (1.41 g, 12.41 mmol, 918.75 μL, 50 eq) was added, and the reaction was carried out at 20° C. for 3 hours. The reaction system is directly concentrated under reduced pressure to obtain the crude product compound 007-1 trifluoroacetate, which can be directly used in the next reaction without purification.
LCMS:MS(ESI)m/z:545.2[M+1] +。 LCMS: MS (ESI) m/z: 545.2 [M+1] + .
步骤2：化合物007和008和009和010的合成Step 2: Synthesis of compounds 007 and 008 and 009 and 010
氮气保护，将化合物007-1(0.19g,208.91μmol,1eq,折算为3.2个三氟乙酸盐)溶于二氯甲烷(20mL)，加入N,N'-二异丙基乙胺(270.01mg,2.09mmol,363.89μL,10eq)，然后加入丙烯酰氯(28.36mg,313.37μmol,25.55μL,1.5eq)，-60℃反应0.5小时。向反应体系中加入饱和碳酸氢钠溶液(20mL)，分液。水相使用二氯甲烷(20mL*3)萃取，分液。合并有机相，使用和饱和食盐水(30mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过HPLC(柱子:Phenomenex luna C18 80*40mm*3μm；流动相:[水(0.04％盐酸)-乙腈]；B(乙腈)％:25％-35％,7min)分离，纯化得到构型1(保留时间t＝5min)和构型2(保留时间t＝6.5min)两个纯品。构型1通过SFC(柱子:REGIS(s,s)WHELK-O1(250mm*50mm,10μm)；流动相:[0.1％氨水-甲醇]；B(甲醇)％:62.5％-62.5％,3min)分离，纯化分别得到化合物007(手性柱出峰位置：2.008min)和化合物008(手性柱出峰位置：2.516min)。构型2通过SFC(柱子:REGIS(s,s)WHELK-O1(250mm*50mm,10μm)；流动相:[0.1％氨水-甲醇]；B(甲醇)％:50％-50％,3min)分离，纯化分别得到化合物009(手性柱出峰位置：1.963min)和化合物010(手性柱出峰位置：2.151min)。Under nitrogen protection, compound 007-1 (0.19g, 208.91μmol, 1eq, converted to 3.2 trifluoroacetate) was dissolved in dichloromethane (20mL), and N,N'-diisopropylethylamine (270.01 mg, 2.09mmol, 363.89μL, 10eq), then add acryloyl chloride (28.36mg, 313.37μmol, 25.55μL, 1.5eq), and react at -60°C for 0.5 hours. A saturated sodium bicarbonate solution (20 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (20 mL*3) and separated. The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by HPLC (column: Phenomenex luna C18 80*40mm*3μm; mobile phase: [water (0.04% hydrochloric acid)-acetonitrile]; B (acetonitrile)%: 25%-35%, 7min), and purified to obtain the configuration 1 (retention time t=5min) and configuration 2 (retention time t=6.5min) two pure products. Configuration 1 passed SFC (column: REGIS(s,s) WHELK-O1 (250mm*50mm, 10μm); mobile phase: [0.1% ammonia-methanol]; B (methanol)%: 62.5%-62.5%, 3min) After separation and purification, compound 007 (peak position of chiral column: 2.008 min) and compound 008 (peak position of chiral column: 2.516 min) were obtained respectively. Configuration 2 passes SFC (column: REGIS(s, s) WHELK-O1 (250mm*50mm, 10μm); mobile phase: [0.1% ammonia-methanol]; B (methanol)%: 50%-50%, 3min) After separation and purification, compound 009 (peak position of chiral column: 1.963 min) and compound 010 (peak position of chiral column: 2.151 min) were obtained respectively.
化合物007：手性柱出峰位置：2.008minCompound 007: Chiral column peak position: 2.008min
LCMS:MS(ESI)m/z:599.3[M+1] +。 LCMS: MS (ESI) m/z: 599.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.54(d,J＝5.2Hz,1H),7.76(d,J＝8.0Hz,1H),7.38-7.32(m,2H),6.74(t,J＝8.8Hz,1H),6.68-6.55(m,2H),6.42(d,J＝16.8Hz,1H),6.11-6.04(m,1H),6.01-5.97(m,1H),5.83(d,J＝10.8Hz,1H),4.85-4.69(m,2H),4.59-4.49(m,1H),4.19(d,J＝9.2Hz,1H),4.04-3.89(m,2H),3.74-3.47(m,2H),3.24-3.15(m,1H),2.94-2.88(m,1H),2.48-2.35(m,2H),1.66(s,3H),1.33(d,J＝6.8Hz,3H),1.02(d,J＝6.8Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.54 (d, J = 5.2 Hz, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.38-7.32 (m, 2H), 6.74 (t, J = 8.8Hz, 1H), 6.68-6.55 (m, 2H), 6.42 (d, J = 16.8Hz, 1H), 6.11-6.04 (m, 1H), 6.01-5.97 (m, 1H), 5.83 (d, J = 10.8Hz, 1H), 4.85-4.69 (m, 2H), 4.59-4.49 (m, 1H), 4.19 (d, J = 9.2Hz, 1H), 4.04-3.89 (m, 2H), 3.74-3.47 ( m,2H),3.24-3.15(m,1H),2.94-2.88(m,1H),2.48-2.35(m,2H),1.66(s,3H),1.33(d,J=6.8Hz,3H) , 1.02 (d, J = 6.8 Hz, 3H).
化合物008：手性柱出峰位置：2.516minCompound 008: Chiral column peak position: 2.516min
LCMS:MS(ESI)m/z:599.3[M+1] +。 LCMS: MS (ESI) m/z: 599.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.55(d,J＝5.2Hz,1H),7.80(d,J＝8.0Hz,1H),7.38-7.32(m,2H),6.74(t,J＝8.4Hz,1H),6.68-6.58(m,2H),6.42(dd,J＝18.0,1.2Hz,1H),6.12-6.05(m,1H),6.02-5.98(m,1H),5.83(dd,J＝10.4,1.6Hz,1H),5.35-5.04(m,1H),4.82-4.58(m,1H),4.19(d,J＝8.8Hz,2H),3.94-3.89(m,2H),3.78(d,J＝14.0Hz,1H),3.48-3.38(m,1H),2.92(s,2H),2.48-2.39(m,2H),1.39(d,J＝6.8Hz,3H),1.34(d,J＝6.8Hz,3H),1.01(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.55 (d, J = 5.2 Hz, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.38-7.32 (m, 2H), 6.74 (t, J = 8.4Hz,1H),6.68-6.58(m,2H),6.42(dd,J=18.0,1.2Hz,1H),6.12-6.05(m,1H),6.02-5.98(m,1H),5.83(dd ,J=10.4,1.6Hz,1H),5.35-5.04(m,1H),4.82-4.58(m,1H),4.19(d,J=8.8Hz,2H),3.94-3.89(m,2H), 3.78 (d, J = 14.0Hz, 1H), 3.48-3.38 (m, 1H), 2.92 (s, 2H), 2.48-2.39 (m, 2H), 1.39 (d, J = 6.8 Hz, 3H), 1.34 (d, J=6.8 Hz, 3H), 1.01 (s, 3H).
化合物009：手性柱出峰位置：1.963minCompound 009: Chiral column peak position: 1.963min
LCMS:MS(ESI)m/z:599.3[M+1] +。 LCMS: MS (ESI) m/z: 599.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.58(d,J＝4.8Hz,1H),7.76(d,J＝8.8Hz,1H),7.35(q,J＝6.8Hz,1H),7.05(d,J＝4.8Hz,1H),6.82(d,J＝8.4Hz,1H),6.75(t,J＝8.4Hz,1H),6.70-6.56(m,1H),6.42(d,J＝16.8Hz,1H),6.20(s,1H),5.83(d,J＝10.4Hz,1H),5.62(t,J＝9.6Hz,1H),4.94-4.76(m,2H),4.53-4.42(m,2H),4.07-3.88(m,2H),3.72-3.51(m,2H),3.25-3.15(m,1H),2.95-2.88(m,1H),2.85-2.78(m,1H),2.23(d,J＝14.4Hz,1H),1.31(d,J＝6.8Hz,3H),1.26(s,3H),1.02(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.58 (d, J = 4.8 Hz, 1H), 7.76 (d, J = 8.8 Hz, 1H), 7.35 (q, J = 6.8 Hz, 1H), 7.05 (d ,J=4.8Hz,1H), 6.82(d,J=8.4Hz,1H), 6.75(t,J=8.4Hz,1H), 6.70-6.56(m,1H), 6.42(d,J=16.8Hz ,1H), 6.20 (s, 1H), 5.83 (d, J = 10.4 Hz, 1H), 5.62 (t, J = 9.6 Hz, 1H), 4.94-4.76 (m, 2H), 4.53-4.42 (m, 2H),4.07-3.88(m,2H),3.72-3.51(m,2H),3.25-3.15(m,1H),2.95-2.88(m,1H),2.85-2.78(m,1H),2.23( d, J = 14.4 Hz, 1H), 1.31 (d, J = 6.8 Hz, 3H), 1.26 (s, 3H), 1.02 (s, 3H).
化合物010：手性柱出峰位置：2.151minCompound 010: Peak position of chiral column: 2.151min
LCMS:MS(ESI)m/z:599.3[M+1] +。 LCMS: MS (ESI) m/z: 599.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.60(d,J＝4.8Hz,1H),7.78(d,J＝8.4Hz,1H),7.36(q,J＝7.2Hz,1H),7.07(d,J＝4.4Hz,1H),6.82(d,J＝8.4Hz,1H),6.75(t,J＝8.8Hz,1H),6.68-6.57(m,1H),6.42(d,J＝16.0Hz,1H),6.22(d,J＝10.4Hz,1H),5.83(dd,J＝10.8,1.6Hz,1H),5.65(t,J＝11.2Hz,1H),4.98-4.71(m,1H),4.44(d,J＝11.6Hz,1H),4.35-4.21(m,1H),4.06-3.87(m,3H),3.76-3.50(m,2H),3.05-2.82(m,3H),2.24(d,J＝16.0Hz,1H),1.33(d,J＝6.8Hz,3H),1.26(s,3H),1.03(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.60 (d, J = 4.8 Hz, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.36 (q, J = 7.2 Hz, 1H), 7.07 (d ,J=4.4Hz,1H), 6.82(d,J=8.4Hz,1H), 6.75(t,J=8.8Hz,1H), 6.68-6.57(m,1H), 6.42(d,J=16.0Hz ,1H), 6.22 (d, J = 10.4Hz, 1H), 5.83 (dd, J = 10.8, 1.6 Hz, 1H), 5.65 (t, J = 11.2Hz, 1H), 4.98-4.71 (m, 1H) ,4.44(d,J=11.6Hz,1H),4.35-4.21(m,1H),4.06-3.87(m,3H),3.76-3.50(m,2H),3.05-2.82(m,3H),2.24 (d, J = 16.0 Hz, 1H), 1.33 (d, J = 6.8 Hz, 3H), 1.26 (s, 3H), 1.03 (s, 3H).
实施例5Example 5
Figure PCTCN2021073149-appb-000052
Figure PCTCN2021073149-appb-000052
合成路线：synthetic route:
Figure PCTCN2021073149-appb-000053
Figure PCTCN2021073149-appb-000053
步骤1：化合物011-3的合成Step 1: Synthesis of compound 011-3
氮气保护，将化合物011-1(8g,41.45mmol,1eq)和化合物001-2(6.58g,37.31mmol,0.9eq)溶于N,N-二甲基甲酰胺(120mL)，加入碳酸铯(27.01g,82.91mmol,2eq)，60℃反应12小时。向反应体系中加入水(150mL)，水相使用甲叔醚(300mL*2)萃取，分液。合并有机相，使用饱和食盐水(200mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝3.0％～20.0％)分离得到化合物011-3。Under nitrogen protection, compound 011-1 (8g, 41.45mmol, 1eq) and compound 001-2 (6.58g, 37.31mmol, 0.9eq) were dissolved in N,N-dimethylformamide (120mL), and cesium carbonate ( 27.01g, 82.91mmol, 2eq), react at 60°C for 12 hours. Water (150 mL) was added to the reaction system, and the aqueous phase was extracted with methyl tertiary ether (300 mL*2) and separated. The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by a column passing machine (mobile phase: ethyl acetate/petroleum ether=3.0%-20.0%) to obtain compound 011-3.
LCMS:MS(ESI)m/z:333.1[M+1] +。 LCMS: MS (ESI) m/z: 333.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.33(d,J＝6.0Hz,1H),7.06(d,J＝5.6Hz,1H),4.27(t,J＝4.4Hz,2H),3.97(t,J＝4.8Hz,2H),0.87(s,9H),0.06(s,6H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.33 (d, J = 6.0 Hz, 1H), 7.06 (d, J = 5.6 Hz, 1H), 4.27 (t, J = 4.4 Hz, 2H), 3.97 (t , J=4.8Hz, 2H), 0.87(s, 9H), 0.06(s, 6H).
步骤2：化合物011-4的合成Step 2: Synthesis of compound 011-4
氮气保护，将化合物011-3(10.2g,30.64mmol,1eq)和异丙烯基三氟硼酸钾(4.76g,32.18mmol,1.05eq)溶于1,4-二氧六环(100mL)和水(10mL)，加入碳酸钾(6.35g,45.97mmol,1.5eq)和1,1'-双(二苯基膦基)二茂铁]二氯化钯(448.45mg,612.88μmol,0.02eq)，100℃反应12小时。将两反应体系合并，向反应体系中加入水(150mL)，水相使用乙酸乙酯(200mL*3)萃取，分液。合并有机相，使用饱和食盐水(200mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝2.0％～20.0％)分离得到化合物011-4。Under nitrogen protection, dissolve compound 011-3 (10.2g, 30.64mmol, 1eq) and potassium isopropenyl trifluoroborate (4.76g, 32.18mmol, 1.05eq) in 1,4-dioxane (100mL) and water (10mL), add potassium carbonate (6.35g, 45.97mmol, 1.5eq) and 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (448.45mg, 612.88μmol, 0.02eq), React at 100°C for 12 hours. The two reaction systems were combined, water (150 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (200 mL*3) and separated. The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by a column passing machine (mobile phase: ethyl acetate/petroleum ether=2.0%-20.0%) to obtain compound 011-4.
LCMS:MS(ESI)m/z:339.1[M+1] +。 LCMS: MS (ESI) m/z: 339.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(d,J＝5.6Hz,1H),6.95(d,J＝5.6Hz,1H),5.34(s,1H),5.25(s,1H),4.23(t,J＝4.8Hz,2H),3.97(t,J＝4.8Hz,2H),2.17(s,3H),0.88(s,9H),0.07(s,6H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.49 (d, J = 5.6 Hz, 1H), 6.95 (d, J = 5.6 Hz, 1H), 5.34 (s, 1H), 5.25 (s, 1H), 4.23 (t, J = 4.8 Hz, 2H), 3.97 (t, J = 4.8 Hz, 2H), 2.17 (s, 3H), 0.88 (s, 9H), 0.07 (s, 6H).
步骤3：化合物011-5的合成Step 3: Synthesis of compound 011-5
将化合物011-4(10.1g,29.84mmol,1eq)溶于乙醇(150mL)，加入钯/碳(5g，10％纯度，氢气(60.28mg,29.84mmol,1eq)置换三次，然后氢气氛围下(50Psi)，30℃反应20小时。将反应体系直接通过硅藻土过滤，滤饼使用乙醇(400mL)冲洗，然后滤液减压浓缩得到化合物011-5，该反应体系无纯化，直接用于下一步反应。Compound 011-4 (10.1g, 29.84mmol, 1eq) was dissolved in ethanol (150mL), palladium/carbon (5g, 10% purity, hydrogen (60.28mg, 29.84mmol, 1eq) was replaced three times, and then under a hydrogen atmosphere ( 50Psi) and react for 20 hours at 30°C. The reaction system was directly filtered through diatomaceous earth, the filter cake was washed with ethanol (400 mL), and then the filtrate was concentrated under reduced pressure to obtain compound 011-5. The reaction system was used directly in the next step without purification. reaction.
LCMS:MS(ESI)m/z:311.1[M+1] +。 LCMS: MS (ESI) m/z: 311.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝7.98(d,J＝5.2Hz,1H),6.62(d,J＝5.6Hz,1H),4.12(t,J＝4.8Hz,2H),4.00(t,J＝5.2Hz,2H),3.74(br s,2H),3.09-3.02(m,1H),1.32(s,3H),1.30(s,3H),0.92(s,9H),0.11(s,6H)。 1 H NMR(400MHz, CDCl 3 )δ=7.98(d,J=5.2Hz,1H), 6.62(d,J=5.6Hz,1H), 4.12(t,J=4.8Hz,2H), 4.00(t ,J=5.2Hz,2H),3.74(br s,2H),3.09-3.02(m,1H),1.32(s,3H),1.30(s,3H),0.92(s,9H),0.11(s ,6H).
步骤4：化合物011-6的合成Step 4: Synthesis of compound 011-6
氮气保护，将化合物001-5(3g,14.35mmol,1eq)溶于四氢呋喃(30mL)，0℃加入草酰氯(7.29g,57.41mmol,5.03mL,4eq)的四氢呋喃(6mL)溶液，升温至75℃反应2小时。将反应体系直接减压浓缩，然后加入四氢呋喃(30mL)，0℃下，缓慢加入化合物011-5(4.06g,13.06mmol,0.91eq)的四氢呋喃(1mL)溶液，0℃反应2小时。向反应体系中缓慢加入饱和氯化铵溶液(30mL)和饱和食盐水溶液(30mL)淬灭该反应，水相使用乙酸乙酯(100mL*3)萃取，分液。合并有机相，使用饱和食盐水(50mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。向粗产品中加入石油醚：乙酸乙酯＝3:1的有机相(60mL)，搅拌1小时，过滤，滤饼减压干燥得到化合物011-6。Under nitrogen protection, dissolve compound 001-5 (3g, 14.35mmol, 1eq) in tetrahydrofuran (30mL), add a solution of oxalyl chloride (7.29g, 57.41mmol, 5.03mL, 4eq) in tetrahydrofuran (6mL) at 0°C, and raise the temperature to 75 React at °C for 2 hours. The reaction system was directly concentrated under reduced pressure, and then tetrahydrofuran (30 mL) was added. At 0° C., a solution of compound 011-5 (4.06 g, 13.06 mmol, 0.91 eq) in tetrahydrofuran (1 mL) was slowly added, and the reaction was carried out at 0° C. for 2 hours. The reaction was quenched by slowly adding saturated ammonium chloride solution (30 mL) and saturated saline solution (30 mL) to the reaction system, and the aqueous phase was extracted with ethyl acetate (100 mL*3) and separated. The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was added with petroleum ether: ethyl acetate = 3:1 organic phase (60 mL), stirred for 1 hour, filtered, and the filter cake was dried under reduced pressure to obtain compound 011-6.
LCMS:MS(ESI)m/z:545.1[M+1] +。 LCMS: MS (ESI) m/z: 545.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝9.69(s,1H),8.60(d,J＝5.6Hz,1H),8.06-8.00(m,1H),7.03(s,1H),4.28(t,J＝4.4Hz,2H),4.00(t,J＝4.8Hz,2H),3.40-3.33(m,1H),1.41(d,J＝6.0Hz,6H),0.86(s,9H),0.05(s,6H)。 1 H NMR (400MHz, CDCl 3 ) δ = 9.69 (s, 1H), 8.60 (d, J = 5.6 Hz, 1H), 8.06-8.00 (m, 1H), 7.03 (s, 1H), 4.28 (t, J = 4.4 Hz, 2H), 4.00 (t, J = 4.8 Hz, 2H), 3.40-3.33 (m, 1H), 1.41 (d, J = 6.0 Hz, 6H), 0.86 (s, 9H), 0.05 ( s,6H).
步骤5：化合物011-7的合成Step 5: Synthesis of compound 011-7
氮气保护，将化合物011-6(4.2g,7.70mmol,1eq)溶于四氢呋喃(63mL)，0℃缓慢加入六甲基二硅基胺基钾(1M四氢呋喃溶液,18.48mL,2.4eq)，然后20℃反应2小时。因未反应完全，0℃下，补加六甲基二硅基胺基钾(1M四氢呋喃溶液,2.31mL,0.3eq)，20℃反应2小时。向反应体系中加入饱和氯化铵溶液(50mL)淬灭反应，水相使用乙酸乙酯(3*100mL)萃取，分液。合并有机相，使用饱和食盐水(100mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝5.0％～75.0％)分离纯化得到化合物011-7。Under nitrogen protection, dissolve compound 011-6 (4.2g, 7.70mmol, 1eq) in tetrahydrofuran (63mL), slowly add potassium hexamethyldisilazide (1M tetrahydrofuran solution, 18.48mL, 2.4eq) at 0°C, then React at 20°C for 2 hours. Since the reaction was not complete, potassium hexamethyldisilazide (1M tetrahydrofuran solution, 2.31 mL, 0.3 eq) was added at 0°C, and the reaction was carried out at 20°C for 2 hours. Saturated ammonium chloride solution (50 mL) was added to the reaction system to quench the reaction, and the aqueous phase was extracted with ethyl acetate (3*100 mL) for liquid separation. The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product is separated and purified by a column machine (mobile phase: ethyl acetate/petroleum ether=5.0%-75.0%) to obtain compound 011-7.
LCMS:MS(ESI)m/z:509.1[M+1] +。 LCMS: MS (ESI) m/z: 509.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.62(d,J＝6.0Hz,1H),8.42(br s,1H),8.21(d,J＝6.8Hz,1H),6.86(d,J＝5.6Hz,1H),4.12-4.05(m,2H),3.82-3.74(m,2H),2.82-2.75(m,1H),1.24(d,J＝6.8Hz,3H),1.18(d,J＝6.8Hz,3H),0.77(s,9H),-0.07(s,3H),-0.10(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.62 (d, J = 6.0 Hz, 1H), 8.42 (br s, 1H), 8.21 (d, J = 6.8 Hz, 1H), 6.86 (d, J = 5.6 Hz,1H),4.12-4.05(m,2H),3.82-3.74(m,2H),2.82-2.75(m,1H),1.24(d,J=6.8Hz,3H),1.18(d,J= 6.8Hz, 3H), 0.77 (s, 9H), -0.07 (s, 3H), -0.10 (s, 3H).
步骤6：化合物011-8的合成Step 6: Synthesis of compound 011-8
氮气保护，将化合物011-7(1.2g,2.36mmol,1eq)加入到三氯氧磷(2.05g,13.37mmol,1.24mL,5.67eq)中，然后加入N,N-二异丙基乙胺(1.52g,11.79mmol,2.05mL,5eq)，40℃反应3小时。得到化合物011-8，该反应体系无纯化，直接用于下一步反应。Under nitrogen protection, add compound 011-7 (1.2g, 2.36mmol, 1eq) to phosphorus oxychloride (2.05g, 13.37mmol, 1.24mL, 5.67eq), and then add N,N-diisopropylethylamine (1.52g, 11.79mmol, 2.05mL, 5eq), react at 40°C for 3 hours. Compound 011-8 was obtained, and the reaction system was directly used in the next reaction without purification.
步骤7：化合物011-9的合成Step 7: Synthesis of compound 011-9
氮气保护，0℃下，将化合物011-8(1.2g,2.27mmol,1eq)溶于四氢呋喃(18mL)，加入N,N-二异丙基乙胺(8.82g,68.25mmol,11.89mL,30eq)，然后加入化合物001-9(1.37g,6.82mmol,3eq)，缓慢升温至25℃反应1小时。将反应体系加入到冰水(30mL)中，水相使用乙酸乙酯(50mL*3)萃取，分液。合并有机相，使用饱和食盐水(60mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝30.0％～100.0％)分离，纯化得到化合物011-9。Under nitrogen protection, at 0℃, dissolve compound 011-8 (1.2g, 2.27mmol, 1eq) in tetrahydrofuran (18mL), add N,N-diisopropylethylamine (8.82g, 68.25mmol, 11.89mL, 30eq) ), then compound 001-9 (1.37g, 6.82mmol, 3eq) was added, and the temperature was slowly raised to 25°C to react for 1 hour. The reaction system was added to ice water (30 mL), and the aqueous phase was extracted with ethyl acetate (50 mL*3) and separated. The organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by a column passing machine (mobile phase: ethyl acetate/petroleum ether = 30.0%-100.0%) and purified to obtain compound 011-9.
LCMS:MS(ESI)m/z:691.2[M+1] +。 LCMS: MS (ESI) m/z: 691.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.59(d,J＝5.6Hz,1H),7.74(dd,J＝7.6,3.2Hz,1H),6.85(d,J＝5.6Hz,1H),4.86-4.77(m,1H),4.29-4.20(m,1H),4.10-3.90(m,4H),3.79-3.54(m,3H),3.32-3.07(m,2H),2.78-2.66(m,1H),1.52(s,9H),1.48-1.45(m,3H),1.27-1.23(m,3H),1.15(t,J＝6.8Hz,3H),0.75(d,J＝2.0Hz,9H),-0.10(d,J＝3.2Hz,3H),-0.15(d,J＝9.2Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.59 (d, J = 5.6 Hz, 1H), 7.74 (dd, J = 7.6, 3.2 Hz, 1H), 6.85 (d, J = 5.6 Hz, 1H), 4.86 -4.77 (m, 1H), 4.29-4.20 (m, 1H), 4.10-3.90 (m, 4H), 3.79-3.54 (m, 3H), 3.32-3.07 (m, 2H), 2.78-2.66 (m, 1H),1.52(s,9H),1.48-1.45(m,3H),1.27-1.23(m,3H),1.15(t,J=6.8Hz,3H),0.75(d,J=2.0Hz,9H ), -0.10 (d, J = 3.2 Hz, 3H), -0.15 (d, J = 9.2 Hz, 3H).
步骤8：化合物011-10的合成Step 8: Synthesis of compound 011-10
氮气保护，将化合物011-9(1.1g,1.59mmol,1eq)和化合物001-11(322.53mg,2.07mmol,1.3eq)溶于1,4-二氧六环(22mL)和水(5.5mL)，加入磷酸钾(675.52mg,3.18mmol,2eq)和1,1'-双(二叔丁基膦)二茂铁]二氯化钯(II)(103.71mg,159.12μmol,0.1eq)，80℃反应4小时。向反应体系中加入水(50mL)，水相使用乙酸乙酯(60mL*3)萃取，分液。合并有机相，使用饱和食盐水(60mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：甲醇/二氯甲烷＝0.5％～3.0％)分离，纯化得到化合物011-10。Under nitrogen protection, compound 011-9 (1.1g, 1.59mmol, 1eq) and compound 001-11 (322.53mg, 2.07mmol, 1.3eq) were dissolved in 1,4-dioxane (22mL) and water (5.5mL ), add potassium phosphate (675.52mg, 3.18mmol, 2eq) and 1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (103.71mg, 159.12μmol, 0.1eq), React at 80°C for 4 hours. Water (50 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (60 mL*3) and separated. The organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column passing machine (mobile phase: methanol/dichloromethane=0.5%-3.0%) and purified to obtain compound 011-10.
LCMS:MS(ESI)m/z:767.2[M+1] +。 LCMS: MS (ESI) m/z: 767.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝9.39(d,J＝5.6Hz,1H),8.61(d,J＝6.0Hz,1H),7.86(dd,J＝11.6,9.6 Hz,1H),7.32-7.28(m,1H),7.14(q,J＝7.6Hz,1H),6.90(d,J＝6.0Hz,1H),5.17-4.92(m,1H),4.75(s,1H),4.57-4.47(m,1H),4.34-4.11(m,1H),4.09-3.89(m,4H),3.70-3.66(m,2H),3.39-3.20(m,2H),2.95-2.85(m,1H),1.53(s,9H),1.45(d,J＝6.8Hz,2H),1.30-1.26(m,4H),1.09(dd,J＝6.8,5.2Hz,3H),0.71(d,J＝2.0Hz,9H),-0.16(d,J＝3.6Hz,3H),-0.20(d,J＝9.2Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 9.39 (d, J = 5.6 Hz, 1H), 8.61 (d, J = 6.0 Hz, 1H), 7.86 (dd, J = 11.6, 9.6 Hz, 1H), 7.32 -7.28 (m, 1H), 7.14 (q, J = 7.6 Hz, 1H), 6.90 (d, J = 6.0 Hz, 1H), 5.17-4.92 (m, 1H), 4.75 (s, 1H), 4.57- 4.47(m,1H),4.34-4.11(m,1H),4.09-3.89(m,4H),3.70-3.66(m,2H),3.39-3.20(m,2H),2.95-2.85(m,1H) ), 1.53 (s, 9H), 1.45 (d, J = 6.8 Hz, 2H), 1.30-1.26 (m, 4H), 1.09 (dd, J = 6.8, 5.2 Hz, 3H), 0.71 (d, J = 2.0 Hz, 9H), -0.16 (d, J = 3.6 Hz, 3H), -0.20 (d, J = 9.2 Hz, 3H).
步骤9：化合物011-11的合成Step 9: Synthesis of compound 011-11
氮气保护，将化合物011-10(1.3g,1.70mmol,1eq)溶于四氢呋喃(60mL)，加入四甲基氟化铵(789.40mg,8.48mmol,5eq)，65℃反应5小时。将反应体系直接减压浓缩，然后加入水(20mL)，水相使用乙酸乙酯(20mL*4)萃取，分液，合并有机相，使用饱和食盐水(20mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到化合物011-11。该反应体系无需纯化，可直接用于下一步反应。Under nitrogen protection, compound 011-10 (1.3 g, 1.70 mmol, 1 eq) was dissolved in tetrahydrofuran (60 mL), tetramethyl ammonium fluoride (789.40 mg, 8.48 mmol, 5 eq) was added, and reacted at 65° C. for 5 hours. The reaction system was directly concentrated under reduced pressure, and then water (20 mL) was added. The aqueous phase was extracted with ethyl acetate (20 mL*4), separated, and the organic phases were combined, washed with saturated brine (20 mL), and dried with anhydrous sodium sulfate. Filter and concentrate under reduced pressure to obtain compound 011-11. The reaction system does not need to be purified and can be used directly in the next reaction.
LCMS:MS(ESI)m/z:653.1[M+1] +。 LCMS: MS (ESI) m/z: 653.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.59(d,J＝5.2Hz,1H),7.84(dd,J＝12.0,9.2Hz,1H),7.25-7.22(m,1H),6.93(d,J＝4.8Hz,1H),6.69(d,J＝8.4Hz,1H),6.63(t,J＝8.8Hz,1H),4.71-4.47(m,1H),4.27-4.23(m,2H),4.17-3.98(m,4H),3.84-3.73(m,2H),3.65(s,1H),3.52-3.25(m,2H),2.94-2.93(m,1H),1.52(s,9H),1.31-1.20(m,6H),1.10(dd,J＝6.8,4.0Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.59 (d, J = 5.2 Hz, 1H), 7.84 (dd, J = 12.0, 9.2 Hz, 1H), 7.25-7.22 (m, 1H), 6.93 (d, J = 4.8Hz, 1H), 6.69 (d, J = 8.4Hz, 1H), 6.63 (t, J = 8.8Hz, 1H), 4.71-4.47 (m, 1H), 4.27-4.23 (m, 2H), 4.17-3.98(m,4H),3.84-3.73(m,2H),3.65(s,1H),3.52-3.25(m,2H),2.94-2.93(m,1H),1.52(s,9H), 1.31-1.20 (m, 6H), 1.10 (dd, J=6.8, 4.0 Hz, 3H).
步骤10：化合物011-12的合成Step 10: Synthesis of compound 011-12
氮气保护，将化合物011-11(0.94g,1.44mmol,1eq)溶于四氢呋喃(75mL)，加入三苯基膦(1.51g,5.76mmol,4eq)，20℃加入偶氮二甲酸二乙酯(1.00g,5.76mmol,1.05mL,4eq)，20℃反应15小时。向反应体系中加入水(50mL)，水相使用乙酸乙酯(100mL*3)萃取，分液。合并有机相，使用饱和食盐水(50mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝30.0％～100.0％，甲醇/二氯甲烷＝2.0％～10.0％)分离，纯化得到化合物011-12。Under nitrogen protection, dissolve compound 011-11 (0.94g, 1.44mmol, 1eq) in tetrahydrofuran (75mL), add triphenylphosphine (1.51g, 5.76mmol, 4eq), add diethyl azodicarboxylate ( 1.00g, 5.76mmol, 1.05mL, 4eq), react at 20°C for 15 hours. Water (50 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (100 mL*3) and separated. The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column passing machine (mobile phase: ethyl acetate/petroleum ether=30.0%-100.0%, methanol/dichloromethane=2.0%-10.0%), and compound 011-12 was obtained by purification.
LCMS:MS(ESI)m/z:635.2[M+1] +。 LCMS: MS (ESI) m/z: 635.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(dd,J＝5.6,0.8Hz,1H),7.77(dd,J＝16.8,8.4Hz,1H),7.41-7.36(m,1H),6.90-6.86(m,2H),6.69(dd,J＝6.0,2.0Hz,1H),4.69-4.66(m,1H),4.55(d,J＝12.8Hz,1H),4.40-4.30(m,2H),4.23-4.17(m,2H),4.08-3.97(m,2H),3.47-3.14(m,3H),3.07-3.00(m,1H),1.52(s,9H),1.36-1.34(m,3H),1.29-1.27(m,3H),1.06(dd,J＝6.8,5.6Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.49 (dd, J = 5.6, 0.8 Hz, 1H), 7.77 (dd, J = 16.8, 8.4 Hz, 1H), 7.41-7.36 (m, 1H), 6.90- 6.86 (m, 2H), 6.69 (dd, J = 6.0, 2.0 Hz, 1H), 4.69-4.66 (m, 1H), 4.55 (d, J = 12.8 Hz, 1H), 4.40-4.30 (m, 2H) ,4.23-4.17(m,2H),4.08-3.97(m,2H),3.47-3.14(m,3H),3.07-3.00(m,1H),1.52(s,9H),1.36-1.34(m, 3H), 1.29-1.27 (m, 3H), 1.06 (dd, J=6.8, 5.6 Hz, 3H).
步骤11：化合物011-13的三氟乙酸盐的合成Step 11: Synthesis of trifluoroacetate salt of compound 011-13
氮气保护，将化合物011-12(1.05g,1.65mmol,1eq)溶于二氯甲烷(40mL)，加入三氟乙酸(9.43g,82.72mmol,6.12mL,50eq)，30℃反应2小时。将反应体系直接减压浓缩，得到化合物011-13的三氟乙酸盐。该反应体系无需纯化，可直接用于下一步反应。Under nitrogen protection, compound 011-12 (1.05 g, 1.65 mmol, 1 eq) was dissolved in dichloromethane (40 mL), trifluoroacetic acid (9.43 g, 82.72 mmol, 6.12 mL, 50 eq) was added, and the reaction was carried out at 30° C. for 2 hours. The reaction system was directly concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 011-13. The reaction system does not need to be purified and can be used directly in the next reaction.
LCMS:MS(ESI)m/z:535.2[M+1] +。 LCMS: MS (ESI) m/z: 535.2 [M+1] + .
步骤12：化合物011和012的合成Step 12: Synthesis of compounds 011 and 012
氮气保护，将化合物011-13(2g,1.55mmol,1eq,折合6.6三氟乙酸盐)溶于二氯甲烷(60mL)，加入N,N-二异丙基乙胺(3.21g,24.86mmol,4.33mL,16eq)，然后加入丙烯酰氯(281.28mg,3.11mmol,253.40μL,2eq)，-60℃反应10分钟。向反应体系中加入饱和碳酸氢钠溶液(50mL)，分液。水相使用 二氯甲烷(50mL*3)萃取，分液。合并有机相，使用饱和食盐水(50mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过HPLC(柱子:Phenomenex luna c18 250mm*100mm*10μm；流动相:[水(0.05％盐酸)-乙腈]；乙腈％:15％-45％,20min)分离，纯化得到产物纯品。纯品通过SFC(柱子:DAICEL CHIRALCEL OD(250mm*30mm,10μm)；流动相:[0.1％氨水-乙醇]；乙醇％:50％-50％,15min)分离，纯化分别得到化合物011和012。Under nitrogen protection, compound 011-13 (2g, 1.55mmol, 1eq, equivalent to 6.6 trifluoroacetate) was dissolved in dichloromethane (60mL), and N,N-diisopropylethylamine (3.21g, 24.86mmol) , 4.33mL, 16eq), then add acryloyl chloride (281.28mg, 3.11mmol, 253.40μL, 2eq), and react at -60°C for 10 minutes. A saturated sodium bicarbonate solution (50 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (50 mL*3) and separated. The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by HPLC (column: Phenomenex luna c18 250mm*100mm*10μm; mobile phase: [water (0.05% hydrochloric acid)-acetonitrile]; acetonitrile%: 15%-45%, 20min) and purified to obtain the pure product. The pure product was separated by SFC (column: DAICEL CHIRALCEL OD (250mm*30mm, 10μm); mobile phase: [0.1% ammonia-ethanol]; ethanol%: 50%-50%, 15min) and purified to obtain compounds 011 and 012, respectively.
化合物011：手性柱出峰位置：1.420minCompound 011: Peak position of chiral column: 1.420min
LCMS:MS(ESI)m/z:589.3[M+1] +。 LCMS: MS (ESI) m/z: 589.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.54(d,J＝5.2Hz,1H),7.78(d,J＝8.4Hz,1H),7.43-7.37(m,1H),6.91-6.87(m,2H),6.80(d,J＝4.8Hz,1H),6.69-6.59(m,1H),6.41(dd,J＝16.8,1.6Hz,1H),5.82(dd,J＝10.4,1.2Hz,1H),4.87-4.69(m,2H),4.60(d,J＝13.6Hz,2H),4.49-4.34(m,2H),4.16(t,J＝10.4Hz,1H),4.03-3.87(m,1H),3.76-3.45(m,2H),3.28-3.17(m,1H),3.08-3.01(m,1H),1.67-1.58(m,3H),1.39(d,J＝6.4Hz,3H),1.11(d,J＝6.8Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.54 (d, J = 5.2 Hz, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.43-7.37 (m, 1H), 6.91-6.87 (m, 2H), 6.80 (d, J = 4.8 Hz, 1H), 6.69-6.59 (m, 1H), 6.41 (dd, J = 16.8, 1.6 Hz, 1H), 5.82 (dd, J = 10.4, 1.2 Hz, 1H) ), 4.87-4.69 (m, 2H), 4.60 (d, J = 13.6Hz, 2H), 4.49-4.34 (m, 2H), 4.16 (t, J = 10.4 Hz, 1H), 4.03-3.87 (m, 1H),3.76-3.45(m,2H),3.28-3.17(m,1H),3.08-3.01(m,1H),1.67-1.58(m,3H),1.39(d,J=6.4Hz,3H) , 1.11 (d, J = 6.8 Hz, 3H).
化合物012：手性柱出峰位置：1.719minCompound 012: Chiral column peak position: 1.719min
LCMS:MS(ESI)m/z:589.3[M+1] +。 LCMS: MS (ESI) m/z: 589.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.55(d,J＝5.2Hz,1H),7.82(d,J＝8.0Hz,1H),7.43-7.37(m,1H),6.91-6.87(m,2H),6.82(d,J＝5.6Hz,1H),6.67-6.57(m,1H),6.42(dd,J＝16.8,1.6Hz,1H),5.82(dd,J＝10.4,1.6Hz,1H),5.38-5.10(m,1H),4.78-4.60(m,2H),4.45-4.34(m,2H),4.19-4.09(m,2H),3.99-3.75(m,2H),3.46-3.38(m,1H),3.06-2.92(m,2H),1.40(d,J＝6.8Hz,3H),1.37(d,J＝6.8Hz,3H),1.11(d,J＝5.2Hz,3H)。 1 H NMR (400MHz, CDCl 3 )δ = 8.55 (d, J = 5.2 Hz, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.43-7.37 (m, 1H), 6.91-6.87 (m, 2H), 6.82 (d, J = 5.6 Hz, 1H), 6.67-6.57 (m, 1H), 6.42 (dd, J = 16.8, 1.6 Hz, 1H), 5.82 (dd, J = 10.4, 1.6 Hz, 1H ), 5.38-5.10 (m, 1H), 4.78-4.60 (m, 2H), 4.45-4.34 (m, 2H), 4.19-4.09 (m, 2H), 3.99-3.75 (m, 2H), 3.46-3.38 (m,1H),3.06-2.92(m,2H), 1.40(d,J=6.8Hz,3H), 1.37(d,J=6.8Hz,3H), 1.11(d,J=5.2Hz,3H) .
实施例6Example 6
Figure PCTCN2021073149-appb-000054
Figure PCTCN2021073149-appb-000054
合成路线：synthetic route:
Figure PCTCN2021073149-appb-000055
Figure PCTCN2021073149-appb-000055
步骤1：化合物013-1的合成Step 1: Synthesis of compound 013-1
氮气保护，将化合物001-3(5g,29.30mmol,1eq)和乙烯三氟硼酸钾(5.89g,43.95mmol,1.5eq)溶于二氧六环(108mL)和水(12mL)，加入碳酸铯(28.64g,87.90mmol,3eq)和(2-二环己基膦基-2’,4’,6’-三异丙基-1,1’-联苯基)[2-(2’-氨基-1,1’-联苯)]氯化钯(II)(1.15g,1.47mmol,0.05eq)，90℃反应6小时。向反应体系中加入水(50mL)，水相使用乙酸乙酯(100mL*3)萃取，分液。合并有机相，使用饱和食盐水(100mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝2.0％～10.0％)分离，纯化得到化合物013-1。Under nitrogen protection, dissolve compound 001-3 (5g, 29.30mmol, 1eq) and potassium ethylene trifluoroborate (5.89g, 43.95mmol, 1.5eq) in dioxane (108mL) and water (12mL), add cesium carbonate (28.64g, 87.90mmol, 3eq) and (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl) (2-(2'-amino -1,1'-biphenyl)] palladium(II) chloride (1.15g, 1.47mmol, 0.05eq), reacted at 90°C for 6 hours. Water (50 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (100 mL*3) and separated. The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column passing machine (mobile phase: ethyl acetate/petroleum ether=2.0%-10.0%) and purified to obtain compound 013-1.
LCMS:MS(ESI)m/z:163.1[M+1] +。 LCMS: MS (ESI) m/z: 163.1 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.03(d,J＝4.8Hz,1H),7.01(d,J＝4.8Hz,1H),6.76(dd,J＝17.2,10.8Hz,1H),5.77(dd,J＝17.2,1.2Hz,1H),5.50(dd,J＝11.2,1.2Hz,1H),3.77(br s,2H),3.10-3.01(m,1H),1.33(s,3H),1.32(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.03 (d, J = 4.8 Hz, 1H), 7.01 (d, J = 4.8 Hz, 1H), 6.76 (dd, J = 17.2, 10.8 Hz, 1H), 5.77 (dd,J=17.2,1.2Hz,1H), 5.50(dd,J=11.2,1.2Hz,1H), 3.77(br s,2H), 3.10-3.01(m,1H),1.33(s,3H) ,1.32(s,3H).
步骤2：化合物013-2的合成Step 2: Synthesis of compound 013-2
氮气保护，将化合物001-5(4g,19.14mmol,1eq)溶于四氢呋喃(65mL)，0℃加入草酰氯(9.72g,76.55mmol,6.70mL,4eq)的四氢呋喃(13mL)溶液，升温至75℃反应2小时。将反应体系直接减压浓缩，然后加入四氢呋喃(65mL)，0℃下，缓慢加入化合物013-1(2.30g,14.16mmol,0.74eq)的四氢呋喃(13mL)溶液，0℃反应2小时。向反应体系中缓慢加入饱和氯化铵溶液(50mL)和饱和食盐水溶液(50mL)淬灭该反应，然后加入乙酸乙酯(200mL)，过滤，得到滤饼。滤液分液后，水相使用乙酸乙酯(200mL*4)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液与滤饼混合，减压浓缩得到粗产品。向粗产品中加入甲醇(60mL)打浆搅拌1小时，过滤，滤液减压浓缩得到化合物013-2。Under nitrogen protection, dissolve compound 001-5 (4g, 19.14mmol, 1eq) in tetrahydrofuran (65mL), add a solution of oxalyl chloride (9.72g, 76.55mmol, 6.70mL, 4eq) in tetrahydrofuran (13mL) at 0°C, and raise the temperature to 75 React at °C for 2 hours. The reaction system was directly concentrated under reduced pressure, and then tetrahydrofuran (65 mL) was added. At 0° C., a solution of compound 013-1 (2.30 g, 14.16 mmol, 0.74 eq) in tetrahydrofuran (13 mL) was slowly added, and the reaction was carried out at 0° C. for 2 hours. Slowly add saturated ammonium chloride solution (50 mL) and saturated salt solution (50 mL) to the reaction system to quench the reaction, then add ethyl acetate (200 mL), and filter to obtain a filter cake. After the filtrate was separated, the aqueous phase was extracted with ethyl acetate (200 mL*4), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was mixed with the filter cake, and concentrated under reduced pressure to obtain a crude product. Methanol (60 mL) was added to the crude product to be slurried for 1 hour, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 013-2.
LCMS:MS(ESI)m/z:397.0[M+1] +。 LCMS: MS (ESI) m/z: 397.0 [M+1] + .
1H NMR(400MHz,CD 3OD)δ＝8.64(d,J＝6.4Hz,1H),8.23(d,J＝6.4Hz,1H),8.18(d,J＝7.6Hz,1H),7.12(dd,J＝17.2,10.8Hz,1H),6.50(d,J＝17.6Hz,1H),6.04(d,J＝11.2Hz,1H),3.69-3.62(m,1H),1.46(s,3H),1.44(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.64 (d, J = 6.4 Hz, 1H), 8.23 (d, J = 6.4 Hz, 1H), 8.18 (d, J = 7.6 Hz, 1H), 7.12 ( dd, J = 17.2, 10.8 Hz, 1H), 6.50 (d, J = 17.6 Hz, 1H), 6.04 (d, J = 11.2 Hz, 1H), 3.69-3.62 (m, 1H), 1.46 (s, 3H) ), 1.44(s, 3H).
步骤3：化合物013-3的合成Step 3: Synthesis of compound 013-3
氮气保护，将化合物013-2(4.5g,11.33mmol,1eq)溶于四氢呋喃(60mL)，0℃缓慢加入六甲基二硅基胺基钾(1M四氢呋喃溶液,26.06mL,2.3eq)，然后20℃反应3.5小时。向反应体系中加入饱和氯化铵溶液(200mL)淬灭反应，然后加入乙酸乙酯(200mL)，通过硅藻土过滤，然后分液。水相使用乙酸乙酯(3*200mL)萃取，分液。合并有机相，使用饱和食盐水(200mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝5.0％～50.0％)分离，纯化得到化合物013-3。Under nitrogen protection, dissolve compound 013-2 (4.5g, 11.33mmol, 1eq) in tetrahydrofuran (60mL), slowly add potassium hexamethyldisilazide (1M tetrahydrofuran solution, 26.06mL, 2.3eq) at 0°C, and then React at 20°C for 3.5 hours. A saturated ammonium chloride solution (200 mL) was added to the reaction system to quench the reaction, and then ethyl acetate (200 mL) was added, filtered through Celite, and then separated. The aqueous phase was extracted with ethyl acetate (3*200 mL) and separated. The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column passing machine (mobile phase: ethyl acetate/petroleum ether=5.0%-50.0%) and purified to obtain compound 013-3.
LCMS:MS(ESI)m/z:361.0[M+1] +。 LCMS: MS (ESI) m/z: 361.0 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.72(d,J＝5.2Hz,1H),8.68(br s,1H),8.25(d,J＝6.8Hz,1H),7.43(d,J＝5.2Hz,1H),6.37(dd,J＝17.6,11.2Hz,1H),5.91(d,J＝17.6Hz,1H),5.46(d,J＝11.2Hz,1H),2.79-2.69(m,1H),1.24(d,J＝6.8Hz,3H),1.15(d,J＝6.4Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.72 (d, J = 5.2 Hz, 1H), 8.68 (br s, 1H), 8.25 (d, J = 6.8 Hz, 1H), 7.43 (d, J = 5.2 Hz, 1H), 6.37 (dd, J = 17.6, 11.2 Hz, 1H), 5.91 (d, J = 17.6 Hz, 1H), 5.46 (d, J = 11.2 Hz, 1H), 2.79-2.69 (m, 1H) ), 1.24 (d, J = 6.8 Hz, 3H), 1.15 (d, J = 6.4 Hz, 3H).
步骤4：化合物013-4的合成Step 4: Synthesis of compound 013-4
氮气保护，将三氯氧磷(6.46g,42.13mmol,3.92mL,5.07eq)加入到化合物013-3(3g,8.32mmol,1eq)中，然后加入二异丙基乙胺(5.37g,41.58mmol,7.24mL,5eq)，40℃反应3小时。减压浓缩后得到化合物013-4，无需纯化，可直接用于下一步反应。Under nitrogen protection, phosphorus oxychloride (6.46g, 42.13mmol, 3.92mL, 5.07eq) was added to compound 013-3 (3g, 8.32mmol, 1eq), and then diisopropylethylamine (5.37g, 41.58 mmol, 7.24 mL, 5 eq), react at 40°C for 3 hours. After concentration under reduced pressure, compound 013-4 is obtained, which can be directly used in the next reaction without purification.
LCMS:MS(ESI)m/z:379.0[M+1] +。 LCMS: MS (ESI) m/z: 379.0 [M+1] + .
步骤5：化合物013-5的合成Step 5: Synthesis of compound 013-5
氮气保护，0℃下，将化合物013-4(3.1g,8.17mmol,1eq)溶于四氢呋喃(50mL)，加入二异丙基乙胺(15.85g,122.62mmol,21.36mL,15eq)，然后加入化合物001-9(2.46g,12.26mmol,1.5eq)，缓 慢升温至25℃反应1小时。因未反应完全，25℃反应14小时。将反应体系加入到冰水(50mL)中，水相使用乙酸乙酯(50mL*3)萃取，分液。合并有机相，使用饱和食盐水(100mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：乙酸乙酯/石油醚＝5.0％～60.0％)分离，纯化得到化合物013-5。Under nitrogen protection, at 0°C, compound 013-4 (3.1g, 8.17mmol, 1eq) was dissolved in tetrahydrofuran (50mL), diisopropylethylamine (15.85g, 122.62mmol, 21.36mL, 15eq) was added, and then added Compound 001-9 (2.46g, 12.26mmol, 1.5eq) was slowly heated to 25°C and reacted for 1 hour. Because the reaction was not complete, the reaction was carried out at 25°C for 14 hours. The reaction system was added to ice water (50 mL), and the aqueous phase was extracted with ethyl acetate (50 mL*3) and separated. The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column passing machine (mobile phase: ethyl acetate/petroleum ether=5.0%-60.0%) and purified to obtain compound 013-5.
LCMS:MS(ESI)m/z:543.3[M+1] +。 LCMS: MS (ESI) m/z: 543.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.67(d,J＝5.2Hz,1H),7.78(d,J＝8.0Hz,1H),7.41(dd,J＝5.2,2.8Hz,1H),6.37-6.26(m,1H),5.87(dd,J＝17.6,7.6Hz,1H),5.36(dd,J＝10.8,6.8Hz,1H),4.89-4.80(m,1H),4.26(s,2H),3.97(s,1H),3.66(br s,1H),3.31-3.09(m,2H),2.68-2.57(m,1H),1.52(s,9H),1.49-1.47(m,3H),1.24(dd,J＝6.4,4.0Hz,3H),1.12(t,J＝6.4Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.67 (d, J = 5.2 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.41 (dd, J = 5.2, 2.8 Hz, 1H), 6.37 -6.26(m,1H), 5.87(dd,J=17.6,7.6Hz,1H), 5.36(dd,J=10.8,6.8Hz,1H), 4.89-4.80(m,1H), 4.26(s,2H ), 3.97 (s, 1H), 3.66 (br s, 1H), 3.31-3.09 (m, 2H), 2.68-2.57 (m, 1H), 1.52 (s, 9H), 1.49-1.47 (m, 3H) , 1.24 (dd, J = 6.4, 4.0 Hz, 3H), 1.12 (t, J = 6.4 Hz, 3H).
步骤6：化合物013-6的合成Step 6: Synthesis of compound 013-6
氮气保护，将化合物013-5(1.6g,2.95mmol,1eq)和化合物001-11(689.11mg,4.42mmol,1.5eq)溶于二氧六环(48mL)和水(12mL)，加入磷酸钾(1.25g,5.89mmol,2eq)和1,1'-双(二叔丁基膦)二茂铁]二氯化钯(II)(192.03mg,294.64μmol,0.1eq)，80℃反应5小时。向反应体系中加入水(50mL)，水相使用乙酸乙酯(60mL*3)萃取，分液。合并有机相，使用饱和食盐水(60mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：甲醇/二氯甲烷＝0.5％～3.0％)分离，纯化得到化合物013-6。Under nitrogen protection, compound 013-5 (1.6g, 2.95mmol, 1eq) and compound 001-11 (689.11mg, 4.42mmol, 1.5eq) were dissolved in dioxane (48mL) and water (12mL), and potassium phosphate was added (1.25g, 5.89mmol, 2eq) and 1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (192.03mg, 294.64μmol, 0.1eq), react at 80℃ for 5 hours . Water (50 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (60 mL*3) and separated. The organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column passing machine (mobile phase: methanol/dichloromethane=0.5%-3.0%) and purified to obtain compound 013-6.
LCMS:MS(ESI)m/z:619.2[M+1] +。 LCMS: MS (ESI) m/z: 619.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝9.18(d,J＝7.2Hz,1H),8.73(d,J＝5.2Hz,1H),7.91(dd,J＝9.6,6.8Hz,1H),7.48(dd,J＝4.8,1.6Hz,1H),7.31-7.25(m,1H),6.71-6.65(m,2H),6.40-6.28(m,1H),5.92(dd,J＝17.2,7.6Hz,1H),5.38(dd,J＝10.8,8.8Hz,1H),5.04-4.84(m,1H),4.48(s,1H),4.30(d,J＝12.4Hz,1H),4.02(s,2H),3.81-3.65(m,2H),3.50-3.01(m,3H),2.88-2.79(m,1H),1.53(s,9H),1.27(dd,J＝6.8,3.6Hz,3H),1.07(t,J＝6.0Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 9.18 (d, J = 7.2 Hz, 1H), 8.73 (d, J = 5.2 Hz, 1H), 7.91 (dd, J = 9.6, 6.8 Hz, 1H), 7.48 (dd,J=4.8,1.6Hz,1H),7.31-7.25(m,1H),6.71-6.65(m,2H),6.40-6.28(m,1H),5.92(dd,J=17.2,7.6Hz ,1H),5.38(dd,J=10.8,8.8Hz,1H),5.04-4.84(m,1H),4.48(s,1H), 4.30(d,J=12.4Hz,1H),4.02(s, 2H),3.81-3.65(m,2H),3.50-3.01(m,3H),2.88-2.79(m,1H),1.53(s,9H),1.27(dd,J=6.8,3.6Hz,3H) , 1.07 (t, J = 6.0 Hz, 3H).
步骤7：化合物013-8的合成Step 7: Synthesis of compound 013-8
氮气保护，将化合物013-6(0.5g,808.18μmol,1eq)和化合物013-7(2.65g,16.16mmol,166.73μL,20eq)溶于乙腈(20mL)，加入碘化钾(402.47mg,2.42mmol,3eq)和碳酸钾(335.10mg,2.42mmol,3eq)，80℃反应15小时。因未反应完全，补加化合物013-7(1.33g,8.08mmol,10eq)、碘化钾(268.32mg,1.62mmol,2eq)和碳酸钾(223.39mg,1.62mmol,2eq)，80℃反应15小时。因未反应完全，80℃反应25小时。向反应体系中加入水(50mL)，水相使用乙酸乙酯(100mL*2)萃取，分液。合并有机相，使用饱和食盐水(50mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过过柱机(流动相：甲醇/二氯甲烷＝0.5％～2.0％)分离，纯化得到化合物013-8。Under nitrogen protection, compound 013-6 (0.5g, 808.18μmol, 1eq) and compound 013-7 (2.65g, 16.16mmol, 166.73μL, 20eq) were dissolved in acetonitrile (20mL), and potassium iodide (402.47mg, 2.42mmol, 3eq) and potassium carbonate (335.10mg, 2.42mmol, 3eq) at 80°C for 15 hours. Since the reaction was not complete, compound 013-7 (1.33g, 8.08mmol, 10eq), potassium iodide (268.32mg, 1.62mmol, 2eq) and potassium carbonate (223.39mg, 1.62mmol, 2eq) were added and reacted at 80°C for 15 hours. Because the reaction was not complete, the reaction was conducted at 80°C for 25 hours. Water (50 mL) was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (100 mL*2) and separated. The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by column passing machine (mobile phase: methanol/dichloromethane=0.5% to 2.0%) and purified to obtain compound 013-8.
LCMS:MS(ESI)m/z:687.3[M+1] +。 LCMS: MS (ESI) m/z: 687.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.57(d,J＝5.2Hz,1H),7.81-7.75(m,1H),7.35-7.30(m,2H),6.69(d,J＝8.4Hz,2H),6.40-6.30(m,1H),5.84-5.77(m,1H),5.41-5.31(m,2H),5.06-4.90(m,2H),4.41- 4.27(m,2H),4.18-3.93(m,3H),3.77-3.60(m,1H),3.40-3.12(m,2H),2.76(s,1H),2.29-2.09(m,2H),1.53(s,9H),1.26-1.24(m,6H),1.11-1.05(m,3H),1.00-0.93(m,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.57 (d, J = 5.2 Hz, 1H), 7.81-7.75 (m, 1H), 7.35-7.30 (m, 2H), 6.69 (d, J = 8.4 Hz, 2H), 6.40-6.30 (m, 1H), 5.84-5.77 (m, 1H), 5.41-5.31 (m, 2H), 5.06-4.90 (m, 2H), 4.41- 4.27 (m, 2H), 4.18- 3.93 (m, 3H), 3.77-3.60 (m, 1H), 3.40-3.12 (m, 2H), 2.76 (s, 1H), 2.29-2.09 (m, 2H), 1.53 (s, 9H), 1.26- 1.24 (m, 6H), 1.11-1.05 (m, 3H), 1.00-0.93 (m, 3H).
步骤8：化合物013-9的合成Step 8: Synthesis of compound 013-9
氮气保护，将化合物013-8(0.46g,669.78μmol,1eq)溶于二氯甲烷(70mL)，加入1,3-双(2,4,6-三甲基苯基)-2-(咪唑烷亚基)(二氯苯亚甲基)(三环己基膦)钌(113.73mg,133.96μmol,0.2eq)，45℃反应3小时。将反应体系直接减压浓缩得到粗产品。粗产品通过过柱机(流动相：甲醇/二氯甲烷＝0.2％～2.0％)分离，纯化得到化合物013-9。Under nitrogen protection, compound 013-8 (0.46g, 669.78μmol, 1eq) was dissolved in dichloromethane (70mL), and 1,3-bis(2,4,6-trimethylphenyl)-2-(imidazole Alkylene)(dichlorobenzylidene)(tricyclohexylphosphine)ruthenium (113.73mg, 133.96μmol, 0.2eq), reacted at 45°C for 3 hours. The reaction system was directly concentrated under reduced pressure to obtain a crude product. The crude product was separated through a column machine (mobile phase: methanol/dichloromethane = 0.2% to 2.0%) and purified to obtain compound 013-9.
LCMS:MS(ESI)m/z:659.3[M+1] +。 LCMS: MS (ESI) m/z: 659.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.58-8.52(m,1H),7.78(dd,J＝17.2,8.0Hz,1H),7.38-7.29(m,3H),6.71-6.65(m,2H),6.02-5.83(m,1H),5.55-4.89(m,1H),4.72-4.47(m,2H),4.40-4.24(m,2H),4.09-3.64(m,3H),3.51-2.92(m,3H),1.53(s,9H),1.40-1.34(m,6H),1.09-1.06(m,3H),1.03-1.00(m,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.58-8.52 (m, 1H), 7.78 (dd, J = 17.2, 8.0 Hz, 1H), 7.38-7.29 (m, 3H), 6.71-6.65 (m, 2H) ), 6.02-5.83 (m, 1H), 5.55-4.89 (m, 1H), 4.72-4.47 (m, 2H), 4.40-4.24 (m, 2H), 4.09-3.64 (m, 3H), 3.51-2.92 (m, 3H), 1.53 (s, 9H), 1.40-1.34 (m, 6H), 1.09-1.06 (m, 3H), 1.03-1.00 (m, 3H).
步骤9：化合物013-10的合成Step 9: Synthesis of compound 013-10
将化合物013-9(0.31g,470.60μmol,1eq)溶于甲醇(60mL)，加入钯/碳(0.2g,纯度：10％p，氢气(950.61μg,470.60μmol,1eq)置换三次，然后氢气氛围下(50Psi)，50℃反应20小时。将反应体系直接通过硅藻土过滤，滤液减压浓缩得到粗产品。粗产品通过HPLC(柱子:Phenomenex luna C18250*50mm*10μm；流动相:[水(0.04％盐酸)-乙腈]；乙腈％:40％-60％,10min)分离，纯化得到化合物013-10。Compound 013-9 (0.31g, 470.60μmol, 1eq) was dissolved in methanol (60mL), palladium/carbon (0.2g, purity: 10% p, hydrogen (950.61μg, 470.60μmol, 1eq) was replaced three times, and then hydrogen Atmosphere (50Psi), react at 50°C for 20 hours. The reaction system was directly filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was passed HPLC (column: Phenomenex luna C18250*50mm*10μm; mobile phase: [water] (0.04% hydrochloric acid)-acetonitrile]; acetonitrile%: 40%-60%, 10 min) separation and purification to obtain compound 013-10.
LCMS:MS(ESI)m/z:661.2[M+1] +。 LCMS: MS (ESI) m/z: 661.2 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.55-8.48(m,1H),7.85-7.78(m,1H),7.35-7.29(m,1H),7.00(d,J＝5.6Hz,1H),6.81-6.73(m,2H),4.83-4.51(m,1H),4.36-3.97(m,4H),3.76-3.55(m,2H),3.40-2.93(m,2H),2.78-2.66(m,1H),2.47-2.20(m,1H),1.53(s,9H),1.47(d,J＝6.8Hz,3H),1.23-1.20(m,6H),1.15(d,J＝6.0Hz,3H),1.08-1.05(m,2H),0.99-0.96(m,2H)。 1 H NMR(400MHz, CDCl 3 )δ=8.55-8.48(m,1H), 7.85-7.78(m,1H), 7.35-7.29(m,1H), 7.00(d,J=5.6Hz,1H), 6.81-6.73(m,2H),4.83-4.51(m,1H),4.36-3.97(m,4H),3.76-3.55(m,2H),3.40-2.93(m,2H),2.78-2.66(m ,1H),2.47-2.20(m,1H),1.53(s,9H),1.47(d,J=6.8Hz,3H),1.23-1.20(m,6H),1.15(d,J=6.0Hz, 3H), 1.08-1.05 (m, 2H), 0.99-0.96 (m, 2H).
步骤10：化合物013-11的三氟乙酸盐的合成Step 10: Synthesis of the trifluoroacetate salt of compound 013-11
氮气保护，将化合物013-10(0.22g,332.95μmol,1eq)溶于二氯甲烷(9mL)，加入三氟乙酸(1.90g,16.65mmol,1.23mL,50eq)，20℃反应1小时。将反应体系直接减压浓缩，得到化合物013-11的三氟乙酸盐。该反应体系无需纯化，可直接用于下一步反应。Under nitrogen protection, compound 013-10 (0.22g, 332.95μmol, 1eq) was dissolved in dichloromethane (9mL), trifluoroacetic acid (1.90g, 16.65mmol, 1.23mL, 50eq) was added and reacted at 20°C for 1 hour. The reaction system was directly concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 013-11. The reaction system does not need to be purified and can be used directly in the next reaction.
LCMS:MS(ESI)m/z:561.2[M+1] +。 LCMS: MS (ESI) m/z: 561.2 [M+1] + .
步骤11：化合物013,014,015和016的合成Step 11: Synthesis of compounds 013, 014, 015 and 016
氮气保护，将化合物013-11(0.3g,295.06μmol,1eq,折合4个三氟乙酸盐)溶于二氯甲烷(15mL)，加入二异丙基乙胺(610.15mg,4.72mmol,822.30μL,16eq)，然后加入丙烯酰氯(53.41mg,590.13μmol,48.12μL,2eq)，-60℃反应10分钟。向反应体系中加入饱和碳酸氢钠溶液(20mL)，分液。水相使用二氯甲烷(50mL*3)萃取，分液。合并有机相，使用和饱和食盐水(50mL)清洗，无水硫酸钠干燥，过滤，减压浓缩得到粗产品。粗产品通过HPLC(柱子:Welch Xtimate C18 100*25mm*3μm；流动相:[水(0.05％盐酸)-乙腈]；乙腈％:15％-35％,8min)分离，纯化得到产物纯品。纯品通过SFC(柱子:DAICEL  CHIRALPAK IG(250mm*30mm,10μm)；流动相:[0.1％氨水-乙醇]；乙醇％:55％-55％,min)分离，纯化分别得到化合物013、化合物015、化合物016和混合物A。混合物A再次通过SFC(柱子:DAICEL CHIRALPAK IG(250mm*30mm,10μm)；流动相:[0.1％氨水-乙醇]；乙醇％:45％-45％,min)分离，纯化得到化合物014和化合物013。Under nitrogen protection, compound 013-11 (0.3g, 295.06μmol, 1eq, equivalent to 4 trifluoroacetate) was dissolved in dichloromethane (15mL), and diisopropylethylamine (610.15mg, 4.72mmol, 822.30) μL, 16eq), then add acryloyl chloride (53.41mg, 590.13μmol, 48.12μL, 2eq), and react at -60°C for 10 minutes. Saturated sodium bicarbonate solution (20 mL) was added to the reaction system, and the layers were separated. The aqueous phase was extracted with dichloromethane (50 mL*3) and separated. The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by HPLC (column: Welch Xtimate C18 100*25mm*3μm; mobile phase: [water (0.05% hydrochloric acid)-acetonitrile]; acetonitrile%: 15%-35%, 8min) and purified to obtain the pure product. The pure product was separated by SFC (column: DAICEL CHIRALPAK IG (250mm*30mm, 10μm); mobile phase: [0.1% ammonia-ethanol]; ethanol%: 55%-55%, min) and purified to obtain compound 013 and compound 015, respectively , Compound 016 and Mixture A. Mixture A was separated again by SFC (column: DAICEL CHIRALPAK IG (250mm*30mm, 10μm); mobile phase: [0.1% ammonia-ethanol]; ethanol%: 45%-45%, min), and purified to obtain compound 014 and compound 013 .
013:手性柱出峰位置：1.682min013: Peak position of chiral column: 1.682min
LCMS:MS(ESI)m/z:615.3[M+1] +。 LCMS: MS (ESI) m/z: 615.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.55(d,J＝5.2Hz,1H),7.80(d,J＝8.4Hz,1H),7.32-7.28(m,1H),7.15(d,J＝5.2Hz,1H),6.82-6.77(m,2H),6.63(s,1H),6.42(dd,J＝16.4,1.6Hz,1H),5.83(dd,J＝10.4,1.6Hz,1H),4.87-4.71(m,2H),4.62-4.50(m,1H),4.07-3.89(m,1H),3.78-3.45(m,3H),3.29-3.18(m,1H),2.95-2.88(m,1H),2.51-2.44(m,1H),2.22-2.16(m,1H),1.87-1.80(m,1H),1.66(s,3H),1.44-1.31(m,2H),1.28(d,J＝6.4Hz,4H),1.15(d,J＝6.4Hz,3H),1.07(d,J＝6.4Hz,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.55 (d, J = 5.2 Hz, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.32-7.28 (m, 1H), 7.15 (d, J = 5.2Hz, 1H), 6.82-6.77 (m, 2H), 6.63 (s, 1H), 6.42 (dd, J = 16.4, 1.6 Hz, 1H), 5.83 (dd, J = 10.4, 1.6 Hz, 1H), 4.87-4.71(m,2H),4.62-4.50(m,1H),4.07-3.89(m,1H),3.78-3.45(m,3H),3.29-3.18(m,1H),2.95-2.88(m ,1H),2.51-2.44(m,1H),2.22-2.16(m,1H),1.87-1.80(m,1H),1.66(s,3H),1.44-1.31(m,2H),1.28(d , J = 6.4 Hz, 4H), 1.15 (d, J = 6.4 Hz, 3H), 1.07 (d, J = 6.4 Hz, 3H).
014:手性柱出峰位置：1.796min014: Peak position of chiral column: 1.796min
LCMS:MS(ESI)m/z:615.3[M+1] +。 LCMS: MS (ESI) m/z: 615.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.49(d,J＝4.8Hz,1H),7.83(d,J＝8.0Hz,1H),7.36-7.30(m,1H),7.00(d,J＝4.8Hz,1H),6.78-6.74(m,2H),6.70-6.56(m,1H),6.42(dd,J＝16.8,1.6Hz,1H),5.83(dd,J＝10.4,1.6Hz,1H),5.01-4.79(m,1H),4.76-4.67(m,1H),4.55-4.43(m,1H),4.37-4.31(m,1H),4.07-3.87(m,1H),3.74-3.51(m,2H),3.27-3.13(m,1H),2.71-2.65(m,2H),2.48-2.40(m,1H),1.58-1.55(m,4H),1.26-1.25(m,1H),1.22(s,3H),1.21(s,3H),1.18-1.13(m,2H),0.97(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.49 (d, J = 4.8 Hz, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.36-7.30 (m, 1H), 7.00 (d, J = 4.8Hz, 1H), 6.78-6.74 (m, 2H), 6.70-6.56 (m, 1H), 6.42 (dd, J = 16.8, 1.6 Hz, 1H), 5.83 (dd, J = 10.4, 1.6 Hz, 1H ),5.01-4.79(m,1H),4.76-4.67(m,1H),4.55-4.43(m,1H), 4.37-4.31(m,1H),4.07-3.87(m,1H),3.74-3.51 (m, 2H), 3.27-3.13 (m, 1H), 2.71-2.65 (m, 2H), 2.48-2.40 (m, 1H), 1.58-1.55 (m, 4H), 1.26-1.25 (m, 1H) , 1.22 (s, 3H), 1.21 (s, 3H), 1.18-1.13 (m, 2H), 0.97 (s, 3H).
015:手性柱出峰位置：2.049min015: Peak position of chiral column: 2.049min
LCMS:MS(ESI)m/z:615.3[M+1] +。 LCMS: MS (ESI) m/z: 615.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.58(d,J＝5.2Hz,1H),7.84(d,J＝8.0Hz,1H),7.33-7.29(m,1H),7.21(d,J＝4.4Hz,1H),6.82-6.78(m,2H),6.69-6.57(m,1H),6.43(dd,J＝16.8,1.6Hz,1H),5.83(dd,J＝10.4,1.6Hz,1H),5.41-5.10(m,1H),4.81-4.56(m,1H),4.23-4.09(m,2H),3.99-3.89(m,1H),3.82-3.75(m,2H),3.55-3.36(m,1H),3.02-2.88(m,1H),2.55-2.47(m,1H),2.28-2.21(m,1H),1.90-1.81(m,1H),1.44(s,1H),1.41(d,J＝6.8Hz,3H),1.30(d,J＝6.8Hz,3H),1.26(s,2H),1.16(d,J＝6.4Hz,3H),1.09(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.58 (d, J = 5.2 Hz, 1H), 7.84 (d, J = 8.0 Hz, 1H), 7.33-7.29 (m, 1H), 7.21 (d, J = 4.4Hz, 1H), 6.82-6.78 (m, 2H), 6.69-6.57 (m, 1H), 6.43 (dd, J = 16.8, 1.6 Hz, 1H), 5.83 (dd, J = 10.4, 1.6 Hz, 1H ),5.41-5.10(m,1H),4.81-4.56(m,1H),4.23-4.09(m,2H),3.99-3.89(m,1H),3.82-3.75(m,2H),3.55-3.36 (m,1H),3.02-2.88(m,1H),2.55-2.47(m,1H),2.28-2.21(m,1H),1.90-1.81(m,1H),1.44(s,1H),1.41 (d, J = 6.8 Hz, 3H), 1.30 (d, J = 6.8 Hz, 3H), 1.26 (s, 2H), 1.16 (d, J = 6.4 Hz, 3H), 1.09 (s, 3H).
016:手性柱出峰位置：2.342min016: Peak position of chiral column: 2.342min
LCMS:MS(ESI)m/z:615.3[M+1] +。 LCMS: MS (ESI) m/z: 615.3 [M+1] + .
1H NMR(400MHz,CDCl 3)δ＝8.71(d,J＝5.6Hz,1H),7.92(d,J＝8.0Hz,1H),7.40(d,J＝4.4Hz,1H),7.38-7.34(m,1H),6.79-6.75(m,2H),6.62(s,1H),6.42(dd,J＝16.8,1.6Hz,1H),5.83(dd,J＝10.4,2.0Hz,1H),5.23-4.97(m,1H),4.79-4.76(m,1H),4.53-4.50(m,1H),4.44-4.32(m,2H),4.09-3.91(m,1H),3.76-3.62(m,2H),3.51-3.34(m,1H),3.10-3.00(m,1H),2.66-2.58(m,1H),1.65-1.57(m,2H),1.51-1.46(m,3H),1.34(d,J＝6.8Hz,3H),1.29-1.21(m,5H),1.08(s,3H)。 1 H NMR (400MHz, CDCl 3 ) δ = 8.71 (d, J = 5.6 Hz, 1H), 7.92 (d, J = 8.0 Hz, 1H), 7.40 (d, J = 4.4 Hz, 1H), 7.38-7.34 (m, 1H), 6.79-6.75 (m, 2H), 6.62 (s, 1H), 6.42 (dd, J = 16.8, 1.6 Hz, 1H), 5.83 (dd, J = 10.4, 2.0 Hz, 1H), 5.23-4.97(m,1H),4.79-4.76(m,1H),4.53-4.50(m,1H),4.44-4.32(m,2H),4.09-3.91(m,1H),3.76-3.62(m ,2H),3.51-3.34(m,1H),3.10-3.00(m,1H),2.66-2.58(m,1H),1.65-1.57(m,2H),1.51-1.46(m,3H),1.34 (d, J=6.8 Hz, 3H), 1.29-1.21 (m, 5H), 1.08 (s, 3H).
生物测试数据：Biological test data:
实验例1：MIA-PA-CA-2细胞实验Experimental example 1: MIA-PA-CA-2 cell experiment
1.实验材料：1. Experimental materials:
DMEM培养基，胎牛血清购自Biosera，马血清购自Gibco。CellTiter-Glo(细胞活率化学发光检测试剂)试剂购自Promega。MIA-PA-CA-2细胞系购南京科佰生物科技有限公司。EnVision多标记分析仪(PerkinElmer)。DMEM medium, fetal bovine serum was purchased from Biosera, and horse serum was purchased from Gibco. CellTiter-Glo (cell viability chemiluminescence detection reagent) reagent was purchased from Promega. MIA-PA-CA-2 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd. EnVision multi-label analyzer (PerkinElmer).
2.实验方法：2. Experimental method:
将MIA-PA-CA-2细胞种于白色96孔板中，80μL细胞悬液每孔，其中包含1000个MIA-PA-CA-2细胞。细胞板置于二氧化碳培养箱中过夜培养。MIA-PA-CA-2 cells were planted in a white 96-well plate, 80 μL of cell suspension per well, which contained 1000 MIA-PA-CA-2 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture.
将待测化合物用排枪进行5倍稀释至第8个浓度，即从2mM稀释至26nM，设置双复孔实验。向中间板中加入78μL培养基，再按照对应位置，转移2μL每孔的梯度稀释化合物至中间板，混匀后转移20μL每孔到细胞板中。转移到细胞板中的化合物浓度范围是10μM至0.13nM。细胞板置于二氧化碳培养箱中培养3天。另准备一块细胞板，在加药当天读取信号值作为最大值(下面方程式中Max值)参与数据分析。向此细胞板每孔加入50μL细胞活率化学发光检测试剂，室温孵育10分钟使发光信号稳定。采用多标记分析仪读数。The compound to be tested is diluted 5-fold to the 8th concentration with a discharge gun, that is, diluted from 2mM to 26nM, and a double-well experiment is set up. Add 78 μL of culture medium to the middle plate, and then transfer 2 μL of each well of the serially diluted compound to the middle plate according to the corresponding position. After mixing, transfer 20 μL of each well to the cell plate. The concentration of the compound transferred to the cell plate ranges from 10 μM to 0.13 nM. The cell plate was placed in a carbon dioxide incubator for 3 days. Another cell plate is prepared, and the signal value is read as the maximum value (Max value in the following equation) on the day of drug addition to participate in data analysis. Add 50μL of cell viability chemiluminescence detection reagent to each well of this cell plate, and incubate for 10 minutes at room temperature to stabilize the luminescence signal. Use multi-marker analyzer to read.
3.数据分析：3. Data analysis:
利用方程式(样品-最小值)/(最大值-最小值)*100％将原始数据换算成抑制率，IC 50的值即可通过四参数进行曲线拟合得出(GraphPad Prism软件中"log(抑制剂)/响应--变量作用域"模式得出)。表3提供了本发明的化合物对MIA-PA-CA-2细胞增殖的抑制活性。 Using the equation (sample-minimum)/(maximum-minimum)*100% to convert the original data into inhibition rate, the value of IC 50 can be obtained by curve fitting with four parameters ("log(" in GraphPad Prism software) Inhibitor)/Response-Variable Scope" model derived). Table 3 provides the inhibitory activity of the compounds of the present invention on the proliferation of MIA-PA-CA-2 cells.
表3本发明化合物体外筛选试验结果Table 3 In vitro screening test results of the compounds of the present invention
Figure PCTCN2021073149-appb-000056
Figure PCTCN2021073149-appb-000056
结论：本发明化合物对MIA-PA-CA-2细胞增殖展现出较好的抑制活性，其中化合物005(即计算例1的WX006)对MIA-PA-CA-2的细胞抑制活性IC 50＝2.7nM，显著优于AMG510的体外活性。该结果与计算例1中通过计算化学方法得出的结论(具有较低ΔE能垒差的相同结构化合物WX006与KRAS G12C蛋白的实际结合中，有可能展现与参照化合物AMG510相似的或更优的结合活性)高度吻 合，充分说明了本发明中的计算化学方法的准确性。所以根据本发明中的计算化学方法预测出来的具有较低ΔE能垒差的化合物，将在细胞增殖中展现出与参照化合物AMG510相似的或更优的抑制活性。 Conclusion: The compound of the present invention exhibits a good inhibitory activity on the proliferation of MIA-PA-CA-2 cells, and the compound 005 (ie WX006 of Calculation Example 1) has an IC 50 of inhibitory activity on MIA-PA-CA-2 cells. nM, significantly better than the in vitro activity of AMG510. This result is consistent with the conclusion drawn by computational chemistry method in Calculation Example 1 ( the actual binding of WX006 , a compound of the same structure with a lower ΔE energy barrier, to KRAS G12C protein may exhibit similar or better results than the reference compound AMG510. The binding activity) is highly consistent, which fully demonstrates the accuracy of the computational chemistry method in the present invention. Therefore, compounds with a lower ΔE energy barrier predicted by the computational chemistry method of the present invention will exhibit similar or better inhibitory activity in cell proliferation than the reference compound AMG510.
试验例2：体内药代动力学研究Test Example 2: In vivo pharmacokinetic study
SD小鼠口服及静脉注射受试化合物的药代动力学研究Study on the pharmacokinetics of oral and intravenous injection of test compounds in SD mice
受试化合物与10％二甲基亚砜/60％聚乙二醇400/30％水溶液混合，涡旋并超声，制备得到1mg/mL澄清溶液，微孔滤膜过滤后备用。选取7至10周龄的雄性SD小鼠，每组两只。静脉注射给予候选化合物溶液，AMG510剂量为3mg/kg，化合物003剂量为2mg/kg。口服给予候选化合物溶液，剂量为10mg/kg。收集一定时间的全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件(美国Pharsight公司)计算药代参数。实验结果如表4所示：The test compound was mixed with a 10% dimethyl sulfoxide/60% polyethylene glycol 400/30% aqueous solution, vortexed and sonicated to prepare a 1 mg/mL clear solution, which was filtered with a microporous membrane for use. Select male SD mice aged 7 to 10 weeks, two in each group. The candidate compound solution was administered intravenously, the dose of AMG510 was 3 mg/kg, and the dose of compound 003 was 2 mg/kg. The candidate compound solution was administered orally at a dose of 10 mg/kg. Collect whole blood for a certain period of time, prepare plasma, analyze drug concentration by LC-MS/MS method, and use Phoenix WinNonlin software (Pharsight, USA) to calculate pharmacokinetic parameters. The experimental results are shown in Table 4:
表4受试化合物的药代动力学结果Table 4 Pharmacokinetic results of test compounds
Figure PCTCN2021073149-appb-000057
Figure PCTCN2021073149-appb-000057
结果结论：相同给药剂量下，003化合物口服给药后的***总暴露量、达峰浓度和生物利用度优于参照化合物AMG510，展现优异的药代动力学特性。Result and conclusion: Under the same dosage, the total system exposure, peak concentration and bioavailability of 003 compound after oral administration are better than the reference compound AMG510, showing excellent pharmacokinetic properties.
试验例3：体内药效学研究Test Example 3: In vivo pharmacodynamic study
人胰腺癌Mia PaCa-2细胞裸小鼠皮下移植肿瘤Balb/c Nude小鼠模型的体内药效学研究In vivo pharmacodynamic study of human pancreatic cancer Mia PaCa-2 cells transplanted subcutaneously in nude mice Balb/c Nude mouse model
1.细胞培养和肿瘤组织准备1. Cell culture and tumor tissue preparation
细胞培养：人胰腺癌Mia PaCa-2细胞(ATCC-CRL-1420)体外单层培养，培养条件为DMEM/F12培养基中加20％胎牛血清，1％双抗，37℃5％二氧化碳孵箱培养。一周两次用胰酶-EDTA进行常规消化处理传代。当细胞饱和度为80％-90％，数量到达要求时，收取细胞，计数，重悬于适量PBS中，1:1加入基质胶，获取细胞密度为25×10 6cells/mL的细胞悬液。 Cell culture: Human pancreatic cancer Mia PaCa-2 cells (ATCC-CRL-1420) are cultured in a monolayer in vitro, and the culture conditions are DMEM/F12 medium with 20% fetal bovine serum, 1% double antibody, 37°C and 5% carbon dioxide incubation Box culture. Use pancreatin-EDTA for routine digestion and passage twice a week. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, and resuspended in an appropriate amount of PBS. Matrigel is added 1:1 to obtain a cell suspension with a cell density of 25×10 6 cells/mL .
细胞接种：将0.2mL(5×10 6cells/mouse个)Mia PaCa-2细胞(加基质胶，体积比为1:1)皮下接种于每只小鼠的右后背，肿瘤平均体积达到190mm 3时，根据肿瘤体积进行随机分组，按照表5中的方案开始给药。 Cell inoculation: 0.2mL (5×10 6 cells/mouse) Mia PaCa-2 cells (with matrigel, volume ratio 1:1) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached 190mm At 3 o'clock, random grouping was performed according to tumor volume, and the administration was started according to the schedule in Table 5.
表5实验动物分组及给药方案Table 5 Experimental animal grouping and dosing schedule
Figure PCTCN2021073149-appb-000058
Figure PCTCN2021073149-appb-000058
注：PO代表口服；QD代表每日一次。Note: PO stands for oral administration; QD stands for once a day.
2.肿瘤测量和实验指标2. Tumor measurement and experimental indicators
每周两次用游标卡尺测量肿瘤直径。肿瘤体积的计算公式为：V＝0.5a×b 2，a和b分别表示肿瘤的长径和短径。 The tumor diameter was measured with vernier calipers twice a week. The calculation formula of the tumor volume is: V=0.5a×b 2 , a and b represent the long diameter and short diameter of the tumor, respectively.
化合物的抑瘤疗效用TGI(％)或相对肿瘤增殖率T/C(％)评价。相对肿瘤增殖率T/C(％)＝TRTV/CRTV×100％(TRTV：治疗组RTV；CRTV：阴性对照组RTV)。根据肿瘤测量的结果计算出相对肿瘤体积(relative tumor volume，RTV)，计算公式为RTV＝Vt/V0，其中V0是分组给药时(即D0)测量所得平均肿瘤体积，Vt为某一次测量时的平均肿瘤体积，TRTV与CRTV取同一天数据。The anti-tumor efficacy of the compound was evaluated by TGI (%) or the relative tumor growth rate T/C (%). Relative tumor proliferation rate T/C(%)=TRTV/CRTV×100% (TRTV: treatment group RTV; CRTV: negative control group RTV). The relative tumor volume (RTV) is calculated according to the results of tumor measurement. The calculation formula is RTV=Vt/V0, where V0 is the average tumor volume measured during group administration (that is, D0), and Vt is the time of a certain measurement. The average tumor volume, TRTV and CRTV data on the same day.
TGI(％)，反映肿瘤生长抑制率。TGI(％)＝[(1-(某处理组给药结束时平均瘤体积－该处理组开始给药时平均瘤体积))/(溶剂对照组治疗结束时平均瘤体积－溶剂对照组开始治疗时平均瘤体积)]×100％。TGI (%), reflects the tumor growth inhibition rate. TGI(%)=[(1-(Average tumor volume at the end of a certain treatment group-average tumor volume at the start of the treatment group))/(Average tumor volume at the end of treatment in the solvent control group-start treatment in the solvent control group Average tumor volume at time)]×100%.
3.实验结果3. Experimental results
实验结果如图28、29所示。The experimental results are shown in Figures 28 and 29.
结果表明：给药21天时，溶剂对照组荷瘤鼠的瘤体积达到1572mm 3，待测物化合物003(5mg/kg)、化合物003(10mg/kg)和化合物003(30mg/kg)的肿瘤体积均值分别为46mm 3、111mm 3和14mm 3；T/C分别为2.79％、8.27％和0.90％；TGI分别为110.37％、105.70％和112.71％，在三个浓度下均 能显著地抑制肿瘤生长(p值均小于0.001)。受试物AMG510(5mg/kg)和AMG510(10mg/kg)的肿瘤体积均值分别为229mm 3和86mm 3，T/C为14.90％和5.81％，TGI为97.17％和107.51％，p值均<0.001，同样具有显著的抑瘤作用，且小鼠各个剂量组的体重平稳，无明显不耐受现象。 The results showed that the tumor volume of the tumor-bearing mice in the solvent control group reached 1572 mm 3 after administration for 21 days, and the tumor volume of the test substances compound 003 (5 mg/kg), compound 003 (10 mg/kg) and compound 003 (30 mg/kg) The average values are 46mm 3 , 111mm 3 and 14mm 3 ; T/C are 2.79%, 8.27% and 0.90% respectively; TGI are 110.37%, 105.70% and 112.71%, respectively. All three concentrations can significantly inhibit tumor growth. (P values are all less than 0.001). The average tumor volume of the test substances AMG510 (5mg/kg) and AMG510 (10mg/kg) were 229mm 3 and 86mm 3 , respectively, the T/C was 14.90% and 5.81%, the TGI was 97.17% and 107.51%, and the p values were all < 0.001, also has a significant anti-tumor effect, and the weight of each dose group of mice is stable, and there is no obvious intolerance.
试验例4：体内药效学研究Test Example 4: In vivo pharmacodynamic study
人非小细胞肺癌NCI-H358细胞裸小鼠皮下移植肿瘤Balb/c Nude小鼠模型的体内药效学研究In vivo pharmacodynamic study of human non-small cell lung cancer NCI-H358 cells transplanted subcutaneously in nude mice Balb/c Nude mouse model
1.细胞培养和肿瘤组织准备1. Cell culture and tumor tissue preparation
细胞培养：人非小细胞肺癌NCI-H358体外单层培养，培养条件为DMEM/F12培养基中加20％胎牛血清，1％双抗，37℃5％二氧化碳孵箱培养。一周两次用胰酶-EDTA进行常规消化处理传代。当细胞饱和度为80％-90％，数量到达要求时，收取细胞，计数，重悬于适量PBS中，1:1加入基质胶，获取细胞密度为25×10 6cells/mL的细胞悬液。 Cell culture: Human non-small cell lung cancer NCI-H358 is cultured in a monolayer in vitro. The culture conditions are DMEM/F12 medium plus 20% fetal bovine serum, 1% double antibody, 37°C and 5% carbon dioxide incubator. Use pancreatin-EDTA for routine digestion and passage twice a week. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, and resuspended in an appropriate amount of PBS. Matrigel is added 1:1 to obtain a cell suspension with a cell density of 25×10 6 cells/mL .
细胞接种：将0.2mL(5×10 6cells/mouse个)NCI-H358细胞(加基质胶，体积比为1:1)皮下接种于每只小鼠的右后背，肿瘤平均体积达到100-150mm 3时，根据肿瘤体积进行随机分组，按照表6中的方案开始给药。 Cell inoculation: 0.2mL (5×10 6 cells/mouse) of NCI-H358 cells (with matrigel, volume ratio 1:1) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached 100- At 150 mm 3 , random grouping was performed according to the tumor volume, and the administration was started according to the schedule in Table 6.
表6实验动物分组及给药方案Table 6 Grouping of Experimental Animals and Dosing Scheme
Figure PCTCN2021073149-appb-000059
Figure PCTCN2021073149-appb-000059
注：PO代表口服；QD代表每日一次。Note: PO stands for oral administration; QD stands for once a day.
2.肿瘤测量和实验指标2. Tumor measurement and experimental indicators
每周两次用游标卡尺测量肿瘤直径。肿瘤体积的计算公式为：V＝0.5a×b 2，a和b分别表示肿瘤的长径和短径。 The tumor diameter was measured with vernier calipers twice a week. The calculation formula of the tumor volume is: V=0.5a×b 2 , a and b represent the long diameter and short diameter of the tumor, respectively.
化合物的抑瘤疗效用TGI(％)或相对肿瘤增殖率T/C(％)评价。相对肿瘤增殖率T/C(％)＝TRTV/CRTV×100％(TRTV：治疗组RTV；CRTV：阴性对照组RTV)。根据肿瘤测量的结果计算出相对肿瘤体积(relative tumor volume，RTV)，计算公式为RTV＝Vt/V0，其中V0是分组给药时(即D0)测量所得平均肿瘤体积，Vt为某一次测量时的平均肿瘤体积，TRTV与CRTV取同一天数据。The anti-tumor efficacy of the compound was evaluated by TGI (%) or the relative tumor growth rate T/C (%). Relative tumor proliferation rate T/C(%)=TRTV/CRTV×100% (TRTV: treatment group RTV; CRTV: negative control group RTV). The relative tumor volume (RTV) is calculated according to the results of tumor measurement. The calculation formula is RTV=Vt/V0, where V0 is the average tumor volume measured during group administration (that is, D0), and Vt is the time of a certain measurement. The average tumor volume, TRTV and CRTV data on the same day.
TGI(％)，反映肿瘤生长抑制率。TGI(％)＝[(1-(某处理组给药结束时平均瘤体积－该处理组开始给药时平均瘤体积))/(溶剂对照组治疗结束时平均瘤体积－溶剂对照组开始治疗时平均瘤体积)]×100％。TGI (%), reflects the tumor growth inhibition rate. TGI(%)=[(1-(Average tumor volume at the end of a certain treatment group-average tumor volume at the start of the treatment group))/(Average tumor volume at the end of treatment in the solvent control group-start treatment in the solvent control group Average tumor volume at time)]×100%.
3.实验结果3. Experimental results
实验结果如图30、31所示。The experimental results are shown in Figures 30 and 31.
结果表明：给药28天时，溶剂对照组荷瘤鼠的瘤体积达到559mm 3，待测物化合物003(1.5mg/kg)、化合物003(5mg/kg)和化合物003(15mg/kg)的肿瘤体积均值分别为307mm 3、138mm 3和50mm 3；T/C分别为55％、24％和9％；TGI分别为54％、91％和111％，在三个浓度下均能显著地抑制肿瘤生长(p值均小于0.001)。受试物AMG510(5mg/kg)的肿瘤体积均值为248mm 3，T/C为44％，TGI为68％，p值均<0.001，同样具有显著的抑瘤作用，且小鼠各个剂量组的体重平稳，无明显不耐受现象。 The results showed that the tumor volume of the tumor-bearing mice in the solvent control group reached 559 mm 3 after 28 days of administration, and the tumors of the test substances compound 003 (1.5 mg/kg), compound 003 (5 mg/kg) and compound 003 (15 mg/kg) The volume averages are 307mm 3 , 138mm 3 and 50mm 3 respectively; T/C are 55%, 24% and 9% respectively; TGI are 54%, 91% and 111%, respectively. All three concentrations can significantly inhibit tumors Growth (all p values are less than 0.001). The average tumor volume of the test substance AMG510 (5mg/kg) is 248mm 3 , the T/C is 44%, the TGI is 68%, and the p values are all <0.001. It also has a significant tumor suppressor effect. The weight is stable and there is no obvious intolerance.

Claims (17)

  1. 式(I)所示化合物或其药学上可接受的盐，The compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
    Figure PCTCN2021073149-appb-100001
    Figure PCTCN2021073149-appb-100001
    其中，in,
    R 1选自H、F、Cl、Br、I和CH 3，所述CH 3任选被1、2或3个R a取代； R 1 is selected H, F, Cl, Br, I and CH 3, CH 3 optionally substituted by the two or three R a;
    R 2选自H、F、Cl、Br和I； R 2 is selected from H, F, Cl, Br and I;
    R 3选自H、F、Cl、Br、I和C 1-3烷基，所述C 1-3烷基任选被1、2或3个R b取代； R 3 is selected from H, F, Cl, Br, and C 1-3 alkyl, said C 1-3 alkyl optionally substituted with 1, 2 or 3 R b;
    R 4选自H、F、Cl、Br和I； R 4 is selected from H, F, Cl, Br and I;
    R 5选自C 1-6烷基和环丙基，所述C 1-6烷基和环丙基任选被1、2或3个R c取代； R 5 is selected from C 1-6 alkyl and cyclopropyl, the C 1-6 alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R c ;
    L 1选自-(CH 2) m-、-(CH 2) m-NR 6-、-(CH 2) m-O-、-NR 6-(CH 2) m-NR 6-、-NR 6-(CH 2) m-O-、-O-(CH 2) m-O-和-(CH 2) n-(CH＝CH) p-(CH 2) m-O-，所述-(CH 2) m-、-(CH 2) m-NR 6-、-(CH 2) m-O-、-NR 6-(CH 2) m-NR 6-、-NR 6-(CH 2) m-O-、-O-(CH 2) m-O-和-(CH 2) n-(CH＝CH) p-(CH 2) m-O-任选被1、2或3个R d取代； L 1 is selected from -(CH 2 ) m -, -(CH 2 ) m -NR 6 -, -(CH 2 ) m -O-, -NR 6 -(CH 2 ) m -NR 6 -, -NR 6 -(CH 2 ) m -O-, -O-(CH 2 ) m -O- and -(CH 2 ) n -(CH=CH) p -(CH 2 ) m -O-, the -(CH 2 ) m -, -(CH 2 ) m -NR 6 -, -(CH 2 ) m -O-, -NR 6 -(CH 2 ) m -NR 6 -, -NR 6 -(CH 2 ) m- O-, -O-(CH 2 ) m -O- and -(CH 2 ) n -(CH=CH) p -(CH 2 ) m -O- are optionally substituted by 1, 2 or 3 R d ;
    m选自1、2、3、4、5和6；m is selected from 1, 2, 3, 4, 5 and 6;
    n选自0和1；n is selected from 0 and 1;
    p选自1和2；p is selected from 1 and 2;
    R 6选自H和CH 3； R 6 is selected from H and CH 3 ;
    各R a、R b和R c分别独立地选自H、F、Cl、Br和I； Each R a, R b and R c are each independently selected from H, F, Cl, Br and I;
    R d选自H、F、Cl、Br、I和CH 3； R d is selected from H, F, Cl, Br, I and CH 3 ;
    带“*”碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在。The carbon atom with "*" is a chiral carbon atom and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.
  2. 根据权利要求1所述化合物或其药学上可接受的盐，其选自The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from
    Figure PCTCN2021073149-appb-100002
    Figure PCTCN2021073149-appb-100002
    其中，R 1、R 2、R 3、R 4、R 5和L 1如权利要求1所定义。 Wherein, R 1 , R 2 , R 3 , R 4 , R 5 and L 1 are as defined in claim 1.
  3. 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，R 1选自H。 The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 1 is selected from H.
  4. 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，R 2选自H。 The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 2 is selected from H.
  5. 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，R 3选自CH 3。 The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 3 is selected from CH 3 .
  6. 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，R 4选自F。 The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 4 is selected from F.
  7. 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，R 5选自C 1-4烷基和环丙基，所述C 1-4烷基和环丙基任选被1、2或3个R c取代。 The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 5 is selected from C 1-4 alkyl and cyclopropyl, and the C 1-4 alkyl and cyclopropyl are optionally selected by 1, 2 or 3 R c substitutions.
  8. 根据权利要求7所述化合物或其药学上可接受的盐，其中，R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
    Figure PCTCN2021073149-appb-100003
    所述CH 3、CH 2CH 3、CH 2CH 3CH 3、
    Figure PCTCN2021073149-appb-100004
    任选被1、2或3个R c取代。     The compound or a pharmaceutically acceptable salt thereof according to claim 7, wherein R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
    Figure PCTCN2021073149-appb-100003
    The CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
    Figure PCTCN2021073149-appb-100004
    Optionally substituted with 1, 2 or 3 R c .
  9. 根据权利要求8所述化合物或其药学上可接受的盐，其中，R 5选自CH 3、CH 2CH 3、CH 2CH 3CH 3、
    Figure PCTCN2021073149-appb-100005
    The compound or a pharmaceutically acceptable salt thereof according to claim 8, wherein R 5 is selected from CH 3 , CH 2 CH 3 , CH 2 CH 3 CH 3 ,
    Figure PCTCN2021073149-appb-100005
  10. 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，m选自1、2、3、4和5。The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein m is selected from 1, 2, 3, 4, and 5.
  11. 根据权利要求10所述化合物或其药学上可接受的盐，其中，L 1选自-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 4O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-、-(CH＝CH)-(CH 2) 2-O-和-(CH＝CH)-(CH 2) 3-O-，所述-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 4O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-、-(CH＝CH)-(CH 2) 2-O-和-(CH＝CH)-(CH 2) 3-O-任选被1、2或3个R d取代。 The compound or a pharmaceutically acceptable salt thereof according to claim 10, wherein L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 4 O-, -( CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH=CH)-CH 2 -O-, -(CH=CH)-(CH 2 ) 2 -O- and -(CH=CH)-(CH 2 ) 3 -O- , The -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 4 O-, -(CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O( CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH=CH)-CH 2 -O-, -(CH=CH)-(CH 2 ) 2 -O- and -(CH=CH)-(CH 2 ) 3 -O- are optionally substituted by 1, 2 or 3 Rd .
  12. 根据权利要求11所述化合物或其药学上可接受的盐，其中，L 1选自-(CH 2) 4-、-(CH 2) 5O-、-(CH 2) 4O-、-(CH 2) 3O-、-(CH 2) 3NH-、-O(CH 2) 2O-、-NH(CH 2) 2O-、-O(CH 2) 3O-、-NH(CH 2) 3O-、-(CH＝CH)-CH 2-O-、-(CH＝CH)-(CH 2) 2-O-和-(CH 2) 3CH(CH 3)O-。 The compound or a pharmaceutically acceptable salt thereof according to claim 11, wherein L 1 is selected from -(CH 2 ) 4 -, -(CH 2 ) 5 O-, -(CH 2 ) 4 O-, -( CH 2 ) 3 O-, -(CH 2 ) 3 NH-, -O(CH 2 ) 2 O-, -NH(CH 2 ) 2 O-, -O(CH 2 ) 3 O-, -NH(CH 2 ) 3 O-, -(CH=CH)-CH 2 -O-, -(CH=CH)-(CH 2 ) 2 -O- and -(CH 2 ) 3 CH(CH 3 )O-.
  13. 根据权利要求1～12任意一项所述化合物或其药学上可接受的盐，其选自The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-12, which is selected from
    Figure PCTCN2021073149-appb-100006
    Figure PCTCN2021073149-appb-100006
    其中，in,
    R 1、R 2、R 3、R 4和R 5如权利要求1所定义； R 1 , R 2 , R 3 , R 4 and R 5 are as defined in claim 1;
    m1选自3、4和5；m1 is selected from 3, 4 and 5;
    m2选自1、2和3；m2 is selected from 1, 2 and 3;
    R d如权利要求1所定义。 R d is as defined in claim 1.
  14. 下列所示化合物或其药学上可接受的盐，The following compounds or their pharmaceutically acceptable salts,
    Figure PCTCN2021073149-appb-100007
    Figure PCTCN2021073149-appb-100007
    Figure PCTCN2021073149-appb-100008
    Figure PCTCN2021073149-appb-100008
  15. 根据权利要求14所述化合物或其药学上可接受的盐，其选自The compound according to claim 14 or a pharmaceutically acceptable salt thereof, which is selected from
    Figure PCTCN2021073149-appb-100009
    Figure PCTCN2021073149-appb-100009
    Figure PCTCN2021073149-appb-100010
    Figure PCTCN2021073149-appb-100010
  16. 根据权利要求14所述化合物或其药学上可接受的盐，其选自The compound according to claim 14 or a pharmaceutically acceptable salt thereof, which is selected from
    Figure PCTCN2021073149-appb-100011
    Figure PCTCN2021073149-appb-100011
  17. 根据权利要求1～16任意一项所述的化合物或其药学上可接受的盐在制备与KRAS抑制剂相关疾病的药物中的应用。The use of the compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof in the preparation of drugs for diseases related to KRAS inhibitors.
PCT/CN2021/073149 2020-01-21 2021-01-21 Macrocyclic compound serving as kras inhibitor WO2021147965A1 (en)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
CN202010071528 2020-01-21
CN202010071528.3 2020-01-21
CN202010514548 2020-06-08
CN202010514548.3 2020-06-08
CN202010986192.3 2020-09-18
CN202010986192 2020-09-18

Publications (1)

Publication Number Publication Date
WO2021147965A1 true WO2021147965A1 (en) 2021-07-29

Family

ID=76993171

Family Applications (2)

Application Number Title Priority Date Filing Date
PCT/CN2021/073149 WO2021147965A1 (en) 2020-01-21 2021-01-21 Macrocyclic compound serving as kras inhibitor
PCT/CN2021/073151 WO2021147967A1 (en) 2020-01-21 2021-01-21 Macrocyclic compound serving as kras inhibitor

Family Applications After (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/073151 WO2021147967A1 (en) 2020-01-21 2021-01-21 Macrocyclic compound serving as kras inhibitor

Country Status (2)

Country Link
CN (1) CN115003668A (en)
WO (2) WO2021147965A1 (en)

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022060836A1 (en) 2020-09-15 2022-03-24 Revolution Medicines, Inc. Indole derivatives as ras inhibitors in the treatment of cancer
WO2022060583A1 (en) 2020-09-03 2022-03-24 Revolution Medicines, Inc. Use of sos1 inhibitors to treat malignancies with shp2 mutations
WO2022134773A1 (en) * 2020-12-21 2022-06-30 上海和誉生物医药科技有限公司 Macrocyclic k-ras g12c inhibitor, preparation method therefor and use thereof
US11453683B1 (en) 2019-08-29 2022-09-27 Mirati Therapeutics, Inc. KRas G12D inhibitors
WO2022233316A1 (en) * 2021-05-06 2022-11-10 南京明德新药研发有限公司 Twelve-membered macrocyclic compound
WO2022235864A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors
WO2022235870A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors for the treatment of cancer
US11548888B2 (en) 2019-01-10 2023-01-10 Mirati Therapeutics, Inc. KRas G12C inhibitors
WO2023060253A1 (en) 2021-10-08 2023-04-13 Revolution Medicines, Inc. Ras inhibitors
WO2023114954A1 (en) 2021-12-17 2023-06-22 Genzyme Corporation Pyrazolopyrazine compounds as shp2 inhibitors
US11697657B2 (en) 2019-10-28 2023-07-11 Merck Sharp & Dohme Llc Small molecule inhibitors of KRAS G12C mutant
US11702418B2 (en) 2019-12-20 2023-07-18 Mirati Therapeutics, Inc. SOS1 inhibitors
EP4227307A1 (en) 2022-02-11 2023-08-16 Genzyme Corporation Pyrazolopyrazine compounds as shp2 inhibitors
WO2023172940A1 (en) 2022-03-08 2023-09-14 Revolution Medicines, Inc. Methods for treating immune refractory lung cancer
WO2023240263A1 (en) 2022-06-10 2023-12-14 Revolution Medicines, Inc. Macrocyclic ras inhibitors
US11890285B2 (en) 2019-09-24 2024-02-06 Mirati Therapeutics, Inc. Combination therapies
US11932633B2 (en) 2018-05-07 2024-03-19 Mirati Therapeutics, Inc. KRas G12C inhibitors
WO2024081674A1 (en) 2022-10-11 2024-04-18 Aadi Bioscience, Inc. Combination therapies for the treatment of cancer

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113248521B (en) * 2020-02-11 2023-07-18 上海和誉生物医药科技有限公司 K-RAS G12C inhibitor and preparation method and application thereof
US20230107642A1 (en) 2020-12-18 2023-04-06 Erasca, Inc. Tricyclic pyridones and pyrimidones

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018217651A1 (en) * 2017-05-22 2018-11-29 Amgen Inc. Kras g12c inhibitors and methods of using the same
WO2019213516A1 (en) * 2018-05-04 2019-11-07 Amgen Inc. Kras g12c inhibitors and methods of using the same
US20190374542A1 (en) * 2018-06-12 2019-12-12 Amgen Inc. Kras g12c inhibitors and methods of using the same
WO2020259513A1 (en) * 2019-06-24 2020-12-30 Guangdong Newopp Biopharmaceuticals Co., Ltd. Heterocyclic compounds as inhibitors of kras g12c

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SG11202001499WA (en) * 2017-09-08 2020-03-30 Amgen Inc Inhibitors of kras g12c and methods of using the same
AU2019284472A1 (en) * 2018-06-11 2020-11-26 Amgen Inc. KRAS G12C inhibitors for treating cancer
MX2022005053A (en) * 2019-10-28 2022-05-18 Merck Sharp & Dohme Llc Small molecule inhibitors of kras g12c mutant.
CN113248521B (en) * 2020-02-11 2023-07-18 上海和誉生物医药科技有限公司 K-RAS G12C inhibitor and preparation method and application thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018217651A1 (en) * 2017-05-22 2018-11-29 Amgen Inc. Kras g12c inhibitors and methods of using the same
WO2019213516A1 (en) * 2018-05-04 2019-11-07 Amgen Inc. Kras g12c inhibitors and methods of using the same
US20190374542A1 (en) * 2018-06-12 2019-12-12 Amgen Inc. Kras g12c inhibitors and methods of using the same
WO2020259513A1 (en) * 2019-06-24 2020-12-30 Guangdong Newopp Biopharmaceuticals Co., Ltd. Heterocyclic compounds as inhibitors of kras g12c

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11932633B2 (en) 2018-05-07 2024-03-19 Mirati Therapeutics, Inc. KRas G12C inhibitors
US11548888B2 (en) 2019-01-10 2023-01-10 Mirati Therapeutics, Inc. KRas G12C inhibitors
US11964989B2 (en) 2019-08-29 2024-04-23 Mirati Therapeutics, Inc. KRas G12D inhibitors
US11453683B1 (en) 2019-08-29 2022-09-27 Mirati Therapeutics, Inc. KRas G12D inhibitors
US11890285B2 (en) 2019-09-24 2024-02-06 Mirati Therapeutics, Inc. Combination therapies
US11697657B2 (en) 2019-10-28 2023-07-11 Merck Sharp & Dohme Llc Small molecule inhibitors of KRAS G12C mutant
US11702418B2 (en) 2019-12-20 2023-07-18 Mirati Therapeutics, Inc. SOS1 inhibitors
WO2022060583A1 (en) 2020-09-03 2022-03-24 Revolution Medicines, Inc. Use of sos1 inhibitors to treat malignancies with shp2 mutations
WO2022060836A1 (en) 2020-09-15 2022-03-24 Revolution Medicines, Inc. Indole derivatives as ras inhibitors in the treatment of cancer
WO2022134773A1 (en) * 2020-12-21 2022-06-30 上海和誉生物医药科技有限公司 Macrocyclic k-ras g12c inhibitor, preparation method therefor and use thereof
WO2022235864A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors
WO2022235870A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors for the treatment of cancer
WO2022233316A1 (en) * 2021-05-06 2022-11-10 南京明德新药研发有限公司 Twelve-membered macrocyclic compound
WO2023060253A1 (en) 2021-10-08 2023-04-13 Revolution Medicines, Inc. Ras inhibitors
WO2023114954A1 (en) 2021-12-17 2023-06-22 Genzyme Corporation Pyrazolopyrazine compounds as shp2 inhibitors
EP4227307A1 (en) 2022-02-11 2023-08-16 Genzyme Corporation Pyrazolopyrazine compounds as shp2 inhibitors
WO2023172940A1 (en) 2022-03-08 2023-09-14 Revolution Medicines, Inc. Methods for treating immune refractory lung cancer
WO2023240263A1 (en) 2022-06-10 2023-12-14 Revolution Medicines, Inc. Macrocyclic ras inhibitors
WO2024081674A1 (en) 2022-10-11 2024-04-18 Aadi Bioscience, Inc. Combination therapies for the treatment of cancer

Also Published As

Publication number Publication date
WO2021147967A1 (en) 2021-07-29
CN115003668A (en) 2022-09-02

Similar Documents

Publication Publication Date Title
WO2021147965A1 (en) Macrocyclic compound serving as kras inhibitor
EP3842433B1 (en) Pyridone-pyrimidine derivative acting as krasg12c mutein inhibitor
EP4105211A1 (en) Pyrimidoheterocyclic compounds and application thereof
CN115335372B (en) Octahydropyrazinodiazepine diketoning organisms
CN113993860B (en) Seven-membered heterocyclic derivatives as KRAS G12C mutein inhibitors
WO2021259331A1 (en) Eight-membered n-containing heterocyclic compound
WO2021110168A1 (en) Spiro compound serving as erk inhibitor, and application thereof
CN116888128A (en) Pyrimido aromatic ring compounds
WO2022247757A1 (en) Fluorine-substituted pyrimidopyridine compound and use thereof
WO2022063308A1 (en) Class of 1,7-naphthyridine compounds and application thereof
CN114728966B (en) Pyrrolotriazines as MNK inhibitors
CN114746417B (en) Biaryl compounds as Pan-RAF kinase inhibitors
CN113874379B (en) Tetraheterocyclic compounds as Cdc7 inhibitors
WO2022262782A1 (en) Glutarimide-substituted isoxazole fused ring compound, and application thereof
TWI811656B (en) Pyrimido heterocyclic compounds and application thereof
WO2022233316A1 (en) Twelve-membered macrocyclic compound
WO2022171013A1 (en) Tetrahydroquinazoline compound
WO2024012456A1 (en) Piperazine bridge-substituted heterocyclic pyrimidine compound
WO2022166991A1 (en) Indoline compound
WO2024066984A1 (en) Tricyclic derivative, pharmaceutical composition and use
WO2022247796A1 (en) Use of cyclin-dependent kinase 9 inhibitor
WO2020200314A1 (en) Nitrogen-containing spiro derivative as ret inhibitor

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: 21745199

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: 21745199

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 21745199

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 23/01/2023)

122 Ep: pct application non-entry in european phase

Ref document number: 21745199

Country of ref document: EP

Kind code of ref document: A1