WO2016000581A1 - 螺环芳基磷氧化物和芳基磷硫化物 - Google Patents

螺环芳基磷氧化物和芳基磷硫化物 Download PDF

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WO2016000581A1
WO2016000581A1 PCT/CN2015/082605 CN2015082605W WO2016000581A1 WO 2016000581 A1 WO2016000581 A1 WO 2016000581A1 CN 2015082605 W CN2015082605 W CN 2015082605W WO 2016000581 A1 WO2016000581 A1 WO 2016000581A1
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Prior art keywords
amino
mmol
phenyl
methoxy
group
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PCT/CN2015/082605
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English (en)
French (fr)
Inventor
丁照中
张明会
陈曙辉
刘希乐
朱屹东
范传文
赵保平
张龙
杨莹莹
郑庆梅
郑善松
万海文
胡金清
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南京明德新药研发股份有限公司
齐鲁制药有限公司
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Priority claimed from CN201410317076.7A external-priority patent/CN105330698B/zh
Application filed by 南京明德新药研发股份有限公司, 齐鲁制药有限公司 filed Critical 南京明德新药研发股份有限公司
Priority to EP15814466.7A priority Critical patent/EP3165530B1/en
Priority to PL15814466T priority patent/PL3165530T3/pl
Priority to ES15814466T priority patent/ES2714576T3/es
Priority to US15/322,854 priority patent/US10053477B2/en
Priority to JP2017501154A priority patent/JP6487527B2/ja
Publication of WO2016000581A1 publication Critical patent/WO2016000581A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system
    • C07F9/65583—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system each of the hetero rings containing nitrogen as ring hetero atom
    • 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
    • A61K31/66—Phosphorus compounds
    • 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
    • A61K31/66—Phosphorus compounds
    • A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
    • C07F9/65615—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings containing a spiro condensed ring system of the formula where at least one of the atoms X or Y is a hetero atom, e.g. S

Definitions

  • the present invention relates to a spirocyclic aryl phosphorus oxide or sulfide as an ALK inhibitor.
  • the invention relates to a compound of formula (I), or a pharmaceutically acceptable salt thereof, as an ALK inhibitor.
  • Protein kinases play a dominant regulatory role in almost all types of cellular biological activities. They include proliferation, apoptosis, cytoskeletal rearrangement, differentiation, development, immune response, nervous system function and conduction. In addition, many diseases and/or disorders are associated with abnormalities, abnormalities or disorders of one or more kinases.
  • Anaplastic lymphoma kinase is part of a family of tyrosine kinase (RTKs) receptor proteins.
  • the ALK gene provides instructions for the tyrosine kinase receptor protein to transmit signals from the cell surface to the cell through a process of signal transduction. This process begins when the cell surface kinase is stimulated and then the kinase dimerizes. After dimerization, the kinase is labeled with a phosphate group, a process known as phosphorylation. This process activates the kinase. The activated kinase is capable of transferring the phosphate group to another protein in the cell and continues to deliver phosphorylation to a downstream series of proteins. This signaling pathway is important for many cellular processes, such as cell growth and division (proliferation) or maturation (differentiation).
  • anaplastic lymphoma kinase Although the specific function of anaplastic lymphoma kinase is still unclear, it is generally believed that it can help regulate the proliferation of nerve cells in the early development of nerve cells.
  • Mutation of the anaplastic lymphoma kinase ALK gene is a fundamental unit of the protein - a change in amino acids. In some neuroblastomas, as well as a tumor patient composed of immature neural cells (neuroblasts), at least 16 mutated ALK genes have been identified. Neuroblastoma and other cancers occur due to genetic mutations in key genes that control cell proliferation and differentiation, making cell growth and division uncontrollable and forming tumors.
  • the most common mutation in neuroblastoma is at position 1275, where arginine is replaced by glutamate (written as Arg1275Gln or R1275Q).
  • the Arg1275Gln mutation is found in both hereditary and dissociative neuroblastomas and is the only common ALK gene mutation found in both cases.
  • anaplastic lymphoma kinases no longer require phosphorylation from extracellular stimuli. Thus, the kinase and downstream signaling pathways are uninterrupted (constitutive activation).
  • the constitutive activation of anaplastic lymphoma kinase increases the proliferation of immature neural cells and ultimately leads to neuroblastoma.
  • the rearrangement of the ALK gene on chromosome 2 increases the risk of suffering from other cancers. These rearrangements are somatic mutations, that is, mutations in somatic cells are present in a person's life and only appear when the cells become cancerous.
  • translocation is the exchange of genetic material between chromosome 2 and other chromosomes.
  • ALK anaplastic large cell lymphoma
  • ALK anaplastic large cell lymphoma
  • T cells immune cells
  • t(2,5) The most common translocation in ALCL occurs between chromosomes 2 and 5, called t(2,5).
  • This translocation fused the ALK gene to the NPM gene and formed a fusion protein of NPM-ALK.
  • IMT inflammatory myofibroblastoma
  • IMT is a rare form of cancer characterized by solid tumors composed of inflammatory cells and a myofibroblast that is important for wound healing. About half of the population of IMT is involved in the translocation of the ALK gene.
  • inversion Another type of rearrangement is called inversion, and inversion occurs when chromosome 2 is broken into two parts, and the DNA of the resulting fragment is inverted. And reinserted into the chromosome.
  • Non-small cell lung cancer is one of the most common forms of lung cancer.
  • chromosome 2 is reversed. This reversal fused the ALK gene to another gene called EML4 to form an EML4-ALK fusion protein.
  • EML4 another gene called EML4 to form an EML4-ALK fusion protein.
  • the fusion proteins produced by these rearranged genes have the dual function of anaplastic lymphoma kinase and chaperone proteins.
  • Crizotinib is the first FDA-approved ALK inhibitor for the treatment of ALK-positive lung cancer. Although the initial response to Crizotinib was very effective, most patients relapsed in the first year of treatment due to drug resistance. On April 29, 2014, the FDA approved Ceritinib for the treatment of anaplastic lymphoma kinase (ALK)-positive metastatic non-small cell lung cancer (NSCLC), including patients who are effective and resistant to crizotinib.
  • ALK anaplastic lymphoma kinase
  • NSCLC metastatic non-small cell lung cancer
  • Still other compounds are being used in clinical research to treat cancer, such as alexinib, AP-26113 and the like.
  • Some heterocyclic compounds have also been disclosed for the treatment of various cancers.
  • the patents include WO2014033136, WO2014025128, WO2014006554, WO2014002922, WO2013192512, WO2013177092, WO2013148857, WO2013138210, WO2012139499, WO2012140114.
  • the present invention relates to a series of novel spirocyclic arylphosphine oxides and sulfides which are inhibitors of ALK and mutants thereof and which are useful in the treatment of cancer and other diseases.
  • the novel spirocyclic arylphosphorus oxides and sulfides of the present invention unexpectedly exhibit inhibitory activity against AL26, ALK mutants and EGFR mutant enzymes superior to AP26113; and these compounds are obtained in ALK positive patients with NSCLC cell lines.
  • Both the PDX model and the Crizotinib-resistant PDX model also exhibited better in vivo efficacy than the reference compound AP26113. Therefore, these compounds may provide more effective treatment for diseases caused by abnormal ALK enzymes.
  • T 1 is selected from N or C (R 01 );
  • R 01 and R 02 on T 2 are bonded to the same N to form a 3 to 6 membered ring having 1, 2 or 3 heteroatoms, said hetero atom being selected from the group consisting of O, S and N;
  • D is selected from -N(R 01 )-, -O-, and -S-;
  • R 3 is selected from the group consisting of R 03 , OR 03 and SR 03 ;
  • R 03 is selected from the group consisting of C 1-4 alkyl, C 1-4 haloalkyl and C 3-5 cycloalkyl-(CH 2 ) 0-3 -;
  • Z is selected from N and C(R 4 );
  • X 1 is selected from C(R x1 ) and N;
  • X 2 is selected from C(R x2 ) and N;
  • R x1 , R x2 , R 1 , R 2 and R 4 are each independently selected from H, F, Cl, Br, I, CN, OH, SH, NH 2 , or optionally 1, 2 or 3 halogens, Substituted by hydroxy and/or cyano: C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl-(CH 2 ) 0-3 - and C 3-6 heterocycloalkyl-( CH 2 ) 0-3 - wherein said "hetero" represents 1, 2, or 3 heteroatoms selected from O, S and N;
  • R x1 and R x2 are bonded to the same atom to form a 5- to 6-membered ring having 1, 2 or 3 heteroatoms selected from O, S and N;
  • R p1 and R p2 are each independently selected from the group consisting of H, C 1-4 alkyl and C 1-4 haloalkyl;
  • R p1 and R p2 are bonded to the same P to form a 5- to 6-membered ring having 1, 2 or 3 heteroatoms selected from O, S, N and P; and
  • Z and The locations are interchangeable.
  • the above R 03 is selected from the group consisting of CH 3 , CD 3 , CF 3 , CHF 2 , CH 2 CH 3 , CH(CH 3 ) 2 , —CH 2 CF 3 , —CH 2 CH 2 CF 3 , -CH 2 CH 2 F and
  • R p1 and R p2 are each independently selected from the group consisting of H, CH 3 , CD 3 , CF 3 , CHF 2 , CH 2 CH 3 , CH(CH 3 ) 2 , -CH 2 CF 3 , -CH 2 CH 2 CF 3 and -CH 2 CH 2 F.
  • NR 01 R 02 on the T 2 is selected from the group consisting of NHCH 3 , N(CH 3 ) 2 , N(CH 2 CH 3 ) 2 ,
  • the above D, D 1-4 or T 2 are independently selected from -NH-, -NMe- and -O-; D 1-4 and T 2 may also be selected from -CH(NCH 3 CH 3 )-.
  • the compound or a pharmaceutically acceptable salt thereof has the structure represented by the formula (II):
  • the compound or a pharmaceutically acceptable salt thereof has the structure represented by the formula (III):
  • the above compound or a pharmaceutically acceptable salt thereof which is selected from the group consisting of:
  • Another object of the present invention is to provide a process for the preparation of the above compounds, the preparation route of which is shown in Scheme A or C:
  • Another object of the present invention is to provide a non-small cell lung cancer and other cancers caused by the above compounds in the treatment of ALK and/or EGFR and their mutations, and ROS1, BRAF, c-MET, HER2, KRAS/MEK, PIK3CA, Use of a combination of an inhibitor such as FDFR, DDR2 and/or VEGFR, and a drug for cancer treated in combination with a cytotoxin such as docetaxel or carboplatin.
  • C 1-6 is selected from the group consisting of C 1 , C 2 , C 3 , C 4 , C 5 and C 6 , the number represents a group of carbon atoms;
  • C 3-6 is selected from C 3 , a group consisting of C 4 , C 5 , and C 6 .
  • pharmaceutically acceptable as used herein is intended to mean that those compounds, materials, compositions and/or dosage forms are within the scope of sound medical judgment and are suitable for use in contact with human and animal tissues. Without excessive toxicity, irritation, allergies sexual reactions or other problems or complications are commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salt refers to a salt of a compound of the invention prepared from a compound having a particular substituent found in the present invention and a relatively non-toxic acid or base.
  • a base addition salt can be obtained by contacting a neutral amount of such a compound with a sufficient amount of a base in a neat solution or a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts or similar salts.
  • an acid addition salt can be obtained by contacting a neutral form of such a compound with a sufficient amount of an acid in a neat solution or a suitable inert solvent.
  • pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogencarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and an organic acid salt, such as 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 me
  • the salt is contacted with a base or acid in a conventional manner, and the parent compound is separated, thereby regenerating the neutral form of the compound.
  • the parent form of the compound differs from the form of its various salts by certain physical properties, such as differences in solubility in polar solvents.
  • a "pharmaceutically acceptable salt” is a derivative of a compound of the invention wherein the parent compound is modified by salt formation with an acid or with a base.
  • pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of bases such as amines, alkali metal or organic salts of acid groups such as carboxylic acids, and the like.
  • Pharmaceutically acceptable salts include the conventional non-toxic salts or quaternary ammonium salts of the parent compound, for example salts formed from non-toxic inorganic or organic acids.
  • non-toxic salts include, but are not limited to, those derived from inorganic acids and organic acids selected from the group consisting of 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, Benzenesulfonic acid, benzoic acid, hydrogencarbonate, carbonic acid, citric acid, edetic acid, ethane disulfonic acid, ethanesulfonic acid, fumaric acid, glucoheptose, gluconic acid, glutamic acid, glycolic acid, Hydrobromic acid, hydrochloric acid, hydroiodide, hydroxyl, hydroxynaphthalene, isethionethane, lactic acid, lactose, dodecylsulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, nitric acid, oxalic acid, Pamoic acid, pantothenic acid, phenylacetic acid, phen
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing an acid group or a base by conventional chemical methods.
  • such salts are prepared by reacting these compounds in water or an organic solvent or a mixture of the two via a free acid or base form with a stoichiometric amount of a suitable base or acid.
  • a nonaqueous medium such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is preferred.
  • the compounds provided herein also exist in the form of prodrugs.
  • Prodrugs of the compounds described herein are readily chemically altered under physiological conditions to convert to the compounds of the invention.
  • prodrugs can be converted to the compounds of the invention by chemical or biochemical methods in an in vivo setting.
  • Certain compounds of the invention may exist in unsolvated or solvated forms, including hydrated forms. In general, the solvated forms are equivalent to the unsolvated forms and are included within the scope of the invention. Certain compounds of the invention may exist in polycrystalline or amorphous form.
  • Certain compounds of the invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereomers, geometric isomers and individual isomers are included within the scope of the invention.
  • the compounds of the invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including the cis and trans isomers, the (-)- and (+)-p-enantiomers, the (R)- and (S)-enantiomers, and the diastereomeric a conformation, a (D)-isomer, a (L)-isomer, and a racemic mixture thereof, and other mixtures, such as enantiomerically or diastereomeric enriched mixtures, all of which belong to It is within the scope of the invention.
  • Additional asymmetric carbon atoms may be present in the substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the invention.
  • optically active (R)- and (S)-isomers as well as the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary wherein the resulting mixture of diastereomers is separated and the auxiliary group cleaved to provide pure The desired enantiomer.
  • a salt of a diastereomer is formed with a suitable optically active acid or base, followed by stepping as is known in the art.
  • the diastereomeric resolution is carried out by crystallization or chromatography, and then the pure enantiomer is recovered.
  • the separation of enantiomers and diastereomers is generally accomplished by the use of chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (eg, formation of an amino group from an amine). Formate).
  • the compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound.
  • radiolabeled compounds can be used, such as tritium (3 H), iodine -125 (125 I) or C-14 (14 C). Alterations of all isotopic compositions of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.
  • pharmaceutically acceptable carrier refers to any formulation or carrier medium that is capable of delivering an effective amount of an active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic side effects to the host or patient, including water, oil, Vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, tackifiers, transdermal enhancers and the like. Their formulations are well known to those skilled in the cosmetic or topical pharmaceutical arts. For additional information on vectors, reference is made to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are hereby incorporated by reference.
  • excipient generally refers to the carrier, diluent and/or vehicle required to formulate an effective pharmaceutical composition.
  • an "effective amount” or “therapeutically effective amount” with respect to a pharmaceutical or pharmacologically active agent refers to a sufficient amount of a drug or agent that is non-toxic but that achieves the desired effect.
  • an "effective amount” of an active substance in a composition refers to the amount required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount will vary from person to person, depending on the age and general condition of the recipient, and also on the particular active substance, and a suitable effective amount in a case can be determined by one skilled in the art based on routine experimentation.
  • active ingredient refers to a chemical entity that is effective in treating a target disorder, disease or condition.
  • substituted means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, including variants of heavy hydrogen and hydrogen, as long as the valence of the particular atom is normal and the substituted compound is stable.
  • it means that two hydrogen atoms are substituted.
  • Ketone substitution does not occur on the aryl group.
  • optionally substituted means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemically achievable.
  • any variable eg, R
  • its definition in each case is independent.
  • the group may optionally be substituted with at most two R, and each case has an independent option.
  • combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
  • substituents When a bond of a substituent can be cross-linked to two atoms on a ring, the substituent can be bonded to any atom on the ring.
  • substituents do not indicate which atom is attached to a compound included in the chemical structural formula including but not specifically mentioned, such a substituent may be bonded through any atomic phase thereof. Combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
  • alkyl and heteroalkyl radicals (including what are commonly referred to as alkylene, alkenyl, heteroalkyl, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl and heterocycloalkenyl)
  • R', R", R"', R"" and R""' are each independently preferably hydrogen, a substituted or unsubstituted heteroalkyl group, a substituted or unsubstituted aryl group (for example, an aryl group substituted by 1 to 3 halogens), a substituted or unsubstituted alkyl group, an alkoxy group, or a thioalkyl group Oxyl a group or an aralkyl group.
  • each R group is independently selected, as when more than one R', R", R"' Each of these groups, R"" and R""' groups.
  • R' and R" When R' and R" are attached to the same nitrogen atom, they can form a 5-, 6- or 7-member with the nitrogen atom. ring.
  • -NR'R is intended to include, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl.
  • alkyl is intended to include carbon.
  • a group bonded to a non-hydrogen group such as a haloalkyl group (e.g., -CF 3 , -CH 2 CF 3 ) and an acyl group (e.g., -C(O)CH 3 , -C(O)CF 3 ,- C(O)CH 2 OCH 3 , etc..
  • a non-hydrogen group such as a haloalkyl group (e.g., -CF 3 , -CH 2 CF 3 ) and an acyl group (e.g., -C(O)CH 3 , -C(O)CF 3 ,- C(O)CH 2 OCH 3 , etc.
  • Two substituents on adjacent atoms of the aryl or heteroaryl ring may be optionally substituted with a substituent of the formula -TC(O)-(CRR')qU-, wherein T and U are independently selected From -NR-, -O-, CRR'- or a single bond, q is an integer from 0 to 3.
  • two substituents on adjacent atoms of the aryl or heteroaryl ring may be optionally substituted with a substituent of the formula -A(CH2)r B-, wherein A and B are independently selected From -CRR'-, -O-, -NR-, -S-, -S(O)-, S(O) 2 -, -S(O) 2 NR'- or a single bond, r is 1 to 4 The integer.
  • a single bond on the new ring thus formed can be replaced with a double bond.
  • two substituents on adjacent atoms of the aryl or heteroaryl ring may be optionally substituted with a substituent of the formula -A(CH2)r B-, wherein s and d are each independently An integer selected from 0 to 3, X is -O-, -NR', -S-, -S(O)-, -S(O) 2 - or -S(O) 2 NR'-.
  • the substituents R, R', R" and R"' are each independently preferably selected from hydrogen and substituted or unsubstituted (C 1 -C 6 )alkyl.
  • halo or halogen
  • haloalkyl is intended to include both monohaloalkyl and polyhaloalkyl.
  • halo(C 1 -C 4 )alkyl is intended to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like. Wait.
  • haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl.
  • Alkoxy represents the above alkyl group having a specified number of carbon atoms attached through an oxygen bridge.
  • the C 1-6 alkoxy group includes a C 1 , C 2 , C 3 , C 4 , C 5 and C 6 alkoxy groups.
  • alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentyloxy and S- Pentyloxy.
  • Cycloalkyl includes saturated cyclic groups such as cyclopropyl, cyclobutyl or cyclopentyl.
  • the 3-7 cycloalkyl group includes C 3 , C 4 , C 5 , C 6 and C 7 cycloalkyl groups.
  • Alkenyl includes hydrocarbon chains in a straight or branched configuration wherein one or more carbon-carbon double bonds, such as vinyl and propylene groups, are present at any stable site on the chain.
  • halo or halogen refers to fluoro, chloro, bromo and iodo.
  • heteroatom as used herein includes atoms other than carbon (C) and hydrogen (H), including, for example, oxygen (O), nitrogen (N), sulfur (S), silicon (Si), germanium (Ge), aluminum ( Al) and boron (B) and the like.
  • hetero a radical containing a hetero atom
  • C carbon
  • hydrogen hydrogen
  • Ring means a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocycloalkyl group, a substituted or unsubstituted aryl group or a substituted or unsubstituted heteroaryl group.
  • the so-called ring includes a fused ring.
  • the number of atoms on the ring is usually defined as the number of elements of the ring.
  • “5 to 7-membered ring” means 5 to 7 atoms arranged in a circle. Unless otherwise specified, the ring optionally contains from 1 to 3 heteroatoms.
  • 5- to 7-membered ring includes, for example, phenylpyridine and piperidinyl; on the other hand, the term “5- to 7-membered heterocycloalkyl ring” includes pyridyl and piperidinyl, but does not include phenyl.
  • ring also includes ring systems containing at least one ring, each of which "ring” independently conforms to the above definition.
  • heterocycle or “heterocyclyl” means a stable monocyclic or bicyclic or bicyclic heterocycle which may be saturated, partially unsaturated or unsaturated (aromatic) which contain a carbon atom and 2, 3 or 4 ring heteroatoms independently selected from N, O and S, wherein any of the above heterocyclic rings may be fused to a phenyl ring to form a bicyclic ring.
  • the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p).
  • the nitrogen atom can be substituted or unsubstituted (i.e., N or NR, wherein R is H or other substituents as already defined herein).
  • the heterocyclic ring can be attached to the side groups of any hetero atom or carbon atom to form a stable structure. If the resulting compound is stable, the heterocycles described herein can undergo substitutions at the carbon or nitrogen sites.
  • the nitrogen atom in the heterocycle is optionally quaternized.
  • a preferred embodiment is that when the total number of S and O atoms in the heterocycle exceeds 1, these heteroatoms are not adjacent to each other. Another preferred embodiment is that the total number of S and O atoms in the heterocycle does not exceed one.
  • aromatic heterocyclic group or "heteroaryl” as used herein means a stable 5, 6, or 7 membered monocyclic or bicyclic or aromatic ring of a 7, 8, 9 or 10 membered bicyclic heterocyclic group, It contains carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O and S.
  • the nitrogen atom can be substituted or unsubstituted (i.e., N or NR, wherein R is H or other substituents as already defined herein).
  • the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p). It is worth noting that the total number of S and O atoms on the aromatic heterocycle does not exceed one.
  • Bridged rings are also included in the definition of heterocycles.
  • a bridged ring is formed when one or more atoms (ie, C, O, N, or S) join two non-adjacent carbon or nitrogen atoms.
  • Preferred bridged rings include, but are not limited to, one carbon atom, two carbon atoms, one nitrogen atom, two nitrogen atoms, and one carbon-nitrogen group. worth it Note that a bridge always converts a single ring into a three ring. In the bridged ring, a substituent on the ring can also be present on the bridge.
  • heterocyclic compounds include, but are not limited to, acridinyl, octanoyl, benzimidazolyl, benzofuranyl, benzofuranylfuranyl, benzindenylphenyl, benzoxazolyl, benzimidin Oxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl, oxazolyl, 4aH-carbazolyl, Porphyrin, chroman, chromene, porphyrin-decahydroquinolinyl, 2H, 6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b] Tetrahydrofuranyl, furyl, furfuryl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-carbazolyl, nonenyl,
  • hydrocarbyl or its subordinate concept (such as alkyl, alkenyl, alkynyl, phenyl, etc.) by itself or as part of another substituent means straight-chain, branched or cyclic
  • the hydrocarbon radical or a combination thereof may be fully saturated, mono- or polyunsaturated, may be monosubstituted, disubstituted or polysubstituted, and may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (methine), may include a divalent or polyvalent radical having the specified number of carbon atoms (e.g., C 1 -C 10 represents 1 to 10 carbons).
  • Hydrocarbyl includes, but is not limited to, aliphatic hydrocarbyl groups including chain and cyclic, including but not limited to alkyl, alkenyl, alkynyl groups including, but not limited to, 6-12 members.
  • An aromatic hydrocarbon group such as benzene, naphthalene or the like.
  • alkyl refers to a straight or branched chain of atoms or a combination thereof, which may be fully saturated, unitary or polyunsaturated, and may include divalent and multivalent radicals.
  • saturated hydrocarbon radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, isobutyl, cyclohexyl, (cyclohexyl).
  • a homolog or isomer of a methyl group, a cyclopropylmethyl group, and an atomic group such as n-pentyl, n-hexyl, n-heptyl, n-octyl.
  • the unsaturated alkyl group has one or more double or triple bonds, and examples thereof include, but are not limited to, a vinyl group, a 2-propenyl group, a butenyl group, a crotyl group, a 2-isopentenyl group, and a 2-(butadienyl group). ), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and Structure.
  • heterohydrocarbyl or its subordinate concept (such as heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, etc.), by itself or in combination with another term, means a stable straight chain, branched chain. Or a cyclic hydrocarbon radical or a combination thereof having a number of carbon atoms and at least one heteroatom.
  • heteroalkyl by itself or in conjunction with another term refers to a stable straight chain, branched hydrocarbon radical or combination thereof, having a number of carbon atoms and at least one heteroatom.
  • the heteroatoms are selected from the group consisting of B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized.
  • the heteroatoms B, O, N and S may be located at any internal position of the heterohydrocarbyl group (except where the hydrocarbyl group is attached to the rest of the molecule).
  • Up to two heteroatoms may be consecutive, for example, -CH 2 -NH-OCH 3.
  • alkoxy alkylamino and “alkylthio” (or thioalkoxy) are customary expressions and refer to those alkane which are attached to the remainder of the molecule through an oxygen atom, an amino group or a sulfur atom, respectively.
  • Base group alkoxy
  • cycloalkyl Unless otherwise specified, the terms “cycloalkyl”, “heterocycloalkyl”, “cyclohetero” or subordinates thereof (such as aryl, heteroaryl, aryl, cycloalkyl, heterocycloalkyl, ring) Alkyl, cycloalkenyl, heterocycloalkenyl, cycloalkenyl, cycloalkynyl, heterocycloalkynyl, cycloalkynyl, and the like, by themselves or in combination with other terms, respectively denote a cyclized “hydrocarbyl group", Heterohydrocarbyl” or “hydrocarbyl”.
  • a hetero atom may occupy a position at which the hetero ring is attached to the rest of the molecule.
  • cycloalkyl groups include, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like.
  • heterocyclic groups include 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl and 2-piperazinyl.
  • aryl denotes a polyunsaturated, aromatic hydrocarbon substituent which may be monosubstituted, disubstituted or polysubstituted, which may be monocyclic or polycyclic (preferably 1 to 3 rings), They are fused together or covalently linked.
  • heteroaryl refers to an aryl (or ring) containing one to four heteroatoms. In an illustrative example, the heteroatoms are selected from the group consisting of B, N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a heteroatom.
  • Non-limiting examples of aryl or heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyridyl Azyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxan Azyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thiophene , 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, indolyl, 2-benzimidazolyl, 5-indenyl
  • aryl groups when used in conjunction with other terms (eg, aryloxy, arylthio, aralkyl), include aryl and heteroaryl rings as defined above.
  • aralkyl is intended to include those radicals to which an aryl group is attached to an alkyl group (eg, benzyl, phenethyl, pyridylmethyl, and the like), including wherein the carbon atom (eg, methylene) has been, for example, oxygen.
  • alkyl groups substituted by an atom such as phenoxymethyl, 2-pyridyloxymethyl 3-(1-naphthyloxy)propyl and the like.
  • leaving group refers to a functional group or atom which may be substituted by another functional group or atom by a substitution reaction (for example, an affinity substitution reaction).
  • substituent groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonic acid Esters and the like; acyloxy groups such as acetoxy, trifluoroacetoxy and the like.
  • protecting group includes, but is not limited to, "amino protecting group", “hydroxy protecting group” or “thiol protecting group”.
  • amino protecting group refers to a protecting group suitable for preventing side reactions at the amino nitrogen position.
  • Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, e.g., tert-butoxycarbonyl (Boc) Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1, 1-di -(4'-methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS) and tert-
  • hydroxy protecting group refers to a protecting group suitable for use in preventing hydroxy side reactions.
  • Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and t-butyl groups; acyl groups such as alkanoyl groups (e.g., acetyl); arylmethyl groups such as benzyl (Bn), Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) )and many more.
  • alkyl groups such as methyl, ethyl and t-butyl groups
  • acyl groups such as alkanoyl groups (e.g., acetyl)
  • arylmethyl groups such as benzyl (Bn), Oxybenzyl (PMB), 9-flu
  • some of the compounds of formula (I) can be prepared according to the synthetic methods described in Scheme A, wherein T 1 represents N and T 2 represents NH.
  • PG is an amino protecting group, preferably from BOC, Bn and Cbz.
  • the phosphonate (A1) is reacted with a Grignard reagent, for example, with methylmagnesium bromide in a solvent of tetrahydrofuran or diethyl ether to give phosphorus oxide (A2).
  • a Grignard reagent for example, with methylmagnesium bromide in a solvent of tetrahydrofuran or diethyl ether to give phosphorus oxide (A2).
  • the coupling reaction is carried out under palladium catalysis in a solvent such as DMF, and in the presence of a base such as potassium carbonate to give an arylphosphorus compound A3; A3 can be converted into a phosphorus-sulfur compound by the action of a Lawson reagent.
  • the arylphosphorus oxide or sulfide is reacted with a dichloropyrimidine derivative or other dichloroheterocyclic compound to give compound A4 which can be coupled with an arylamine A7 under acidic conditions, for example in the presence of methanesulfonic acid.
  • the tert-butanol is reacted as a solvent to give the compound A8 of the present invention, or may be further reacted, such as reductive amination, to give a compound of the formula (I).
  • Spirocyclic amino compound A5 can be purchased from a chemical supplier, such as Aldrich Chemical, or can be prepared by Scheme B below.
  • the spirocyclic amino compound A5 is subjected to a substitution reaction in a solvent such as DMF to obtain a compound A6, which is further reduced by hydrogenation to give an arylamine A7.
  • Scheme B is a general synthesis method of spirocyclic amine compound B5 (A5).
  • the ketone B1 is reacted with cyanoacetate to obtain a spiro compound B2, which is hydrolyzed to obtain B3, then B4, and a reducing agent such as lithium aluminum hydride is used to reduce B4.
  • B5 is obtained; B5 can be coupled with the aryl chloride A4 of Scheme A to give A6.
  • Scheme C is a general synthetic procedure for the preparation of compounds of formula (I) wherein T1 is CH.
  • the C1 compound aldehyde is reacted with methyl vinyl ketone under the action of a base such as KOH to obtain ketone C2, hydrogenated to obtain C3, trifluoromethanesulfonate to form C4; and then coupled with aminoaryl or nitroaromatic acid to obtain C5.
  • the invention is now further described by way of examples.
  • the examples given below are for illustrative purposes only and are not intended to be limited to the scope of the invention.
  • Wai. The compounds of the present invention can be prepared by a number of methods known in the art of organic synthesis. Embodiments of the present invention can be synthesized using the methods described below, as well as synthetic methods known in the art of organic synthetic chemistry, or by improved methods based thereon. Preferred methods include, but are not limited to, the methods described below.
  • the present invention employs the following abbreviations: aq for water; DCM for methylene chloride; PE for petroleum ether; DMF for N,N-dimethylformamide; DMSO for dimethyl sulfoxide; EtOAc for ethyl acetate; EtOH stands for ethanol; MeOH stands for methanol; Cbz stands for benzyloxycarbonyl, an amine protecting group; Boc stands for t-butyloxycarbonyl, an amine protecting group; HOAc stands for acetic acid; and NaBH(OAc) 3 stands for triacetoxyborohydride Sodium; rt stands for room temperature; THF stands for tetrahydrofuran; Boc 2 O stands for di-tert-butyldicarbonate; TFA stands for trifluoroacetic acid; DIPEA stands for diisopropylethylamine; Pd(dppf)Cl 2 stands for [1,1'- Bis(diphenylphosphino)ferrocen
  • High performance liquid chromatography was performed using a Shimadzu LC20AB system equipped with a Shimadzu SIL-20A autosampler and a Shimadzu DAD: SPD-M20A detector using a Xtimate C18 (3 m packing, size 2.1 x 300 mm) column.
  • 0-60AB_6 min method Apply a linear gradient, start elution with 100% A (A is 0.0675% TFA in water), and end the elution with 60% B (B is 0.0625% TFA in MeCN solution). The whole process is 4.2 minutes, then eluted with 60% B for 1 minute. The column was equilibrated for 0.8 minutes to reach 100:0 with a total run time of 6 minutes.
  • 10-80AB_6 min method Apply a linear gradient, start elution with 90% A (A is 0.0675% TFA in water), and end the elution with 80% B (B in 0.0625% TFA in acetonitrile). 4.2 minutes, then eluted with 80% B for 1 minute.
  • the column was equilibrated for 0.8 minutes to 90:10 with a total run time of 6 minutes.
  • the column temperature was 50 ° C and the flow rate was 0.8 mL/min.
  • the diode array detector has a scanning wavelength of 200-400 nm.
  • TLC Thin layer chromatography
  • a common solvent for flash column chromatography or thin layer chromatography is a mixture of dichloromethane/methanol, ethyl acetate/methanol and hexane/ethyl acetate.
  • Example 1A A mixture of Example 1A (3.45 g, 9.33 mmol) and a copper-zinc reagent (3.18 g, 64.3 mmol) in anhydrous THF (25.0 mL) was warmed to reflux, and then phosphorus oxychloride (7.78 g, A solution of 50.7 mmol) and 2,2,2-trichloroacetyl chloride (8.75 g, 48.1 mmol) in dry THF (25.0 mL). The reaction mixture was stirred under reflux for 12 hours. TLC (PE) showed that the reaction was completed, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give a brown oily compound (6.0 g, crude product).
  • phosphorus oxychloride 7.78 g, A solution of 50.7 mmol
  • 2,2,2-trichloroacetyl chloride 8.75 g, 48.1 mmol
  • Example 1D To a mixture of Example 1D (0.99 g, 4.27 mmol) in dry THF (25.0 mL), nitromethane (521 mg, 8.54 mmol) and TBAF (1.67 g, 6.41 mmol). The reaction mixture was heated to 70 ° C and stirred for 16 hours. The mixture was cooled to 18 ° C and concentrated to dryness crystals crystals crystals crystals Oily.
  • Example 1E To a solution of Example 1E (1.15 g, 3.92 mmol) in methanol (15.0 mL), EtOAc (EtOAc) The reaction mixture was heated to 50 ° C and stirred in hydrogen (pressure: 50 Psi) for 16 hours. LCMS showed the reaction was completed. The reaction mixture was filtered with EtOAc EtOAcjjjjj LCMS (ESI) (10-80CD): m.
  • Example 1G 240 mg, 1.0 mmol
  • acetic anhydride 5.0 mL
  • concentrated nitric acid 1.0 mL
  • LCMS showed the reaction was complete.
  • the mixture was filtered, and the title crystalljjjjjjj LCMS (ESI) (5-95AB): m/z: 2921.
  • Example 1K To a mixture of Example 1K (2.50 g, 14.8 mmol) and 2,4,5-trichloropyrimidine (2.85 g, 15.5 mmol) in DMF (20 mL), DIPEA (3.82 g, 29.6) Millimoles). The reaction mixture was then heated to 70 ° C and stirred for 16 hours. TLC showed the reaction was complete. The reaction mixture was diluted with water (50 mL) andEtOAc. The combined organic phases were washed with EtOAc EtOAc m.
  • Example 1I 120 mg, 0.49 mmol
  • Example 1L embodiment 154 mg, 0.49 mmol
  • MeSO 3 H 141 mg, 1.47 mmol
  • the reaction mixture was heated to 90 ° C under nitrogen for 12 hours.
  • the title compound (52.2 mg, yield 20%) was obtained as a brown oil.
  • 1 H NMR 400MHz, CDCl 3 ): 3.69-3.56 (m, 4H), 3.47-3.37 (m, 4H), 1.60-1.51 (m, 4H), 1.50-1.43 (m, 9H).
  • Example 5C To a mixture of Example 5C (10.50 g, 40.12 mmol) and methylamine hydrochloride (10.51 g, 155.65 mmol) in methanol (100 mL) was added sodium cyanoborohydride (14.50 g, 230.7). Millimol), then the mixture was stirred at 100 ° C for 16 hours. TLC showed the reaction was complete. The mixture was concentrated to dryness EtOAc EtOAc m.
  • Zinc powder was added to a mixture of Example 5F (1.20 g, 4.12 mmol) and ammonium chloride (1.40 g, 26.17 mmol) in methanol (20 mL) and dichloromethane (2 mL). 2.00 g, 30.59 mmol), then the reaction mixture was stirred at 35 ° C for 10 minutes. TLC showed the reaction was complete. The mixture was filtered and the filtrate was concentrated. The concentrated residue was dissolved in aq.
  • This example was prepared according to the method of Example 4A, replacing the methyl ester of tert-butyl ester 2,7-diazaspiro[3.5]nonane-2-carboxylate with the tert-butyl ester 2,6-diazaspiro[3.4 ] tert-butyl octane-2-carboxylate.
  • the title compound was obtained as a colorless oily compound (yield: 92%).
  • Example 8A A mixture of Example 8A (1.00 g, 3.10 mmol, 1.00 eq.) of sulfuric acid (88%, 4 mL) was stirred at 60 ° C for 4 hr. Water (1.4 mL) was then added to the reaction mixture, and the mixture was heated to 100 ° C and stirred for 1 hour. Water (5 mL) was added to the reaction mixture, which was cooled to 10 ° C and stirred at 10 ° C for 30 min then filtered. The filter cake was washed with EtOAc (EtOAc)EtOAc.
  • EtOAc EtOAc
  • Example 8B To a flask of Example 8B (1.10 g, 3.22 mmol, 1.00 equiv) was added NaOH aqueous solution (5 N, 5 mL) at 15 ° C, then the mixture was warmed to 70 ° C and stirred for 4 hours. The mixture was cooled to 45 ° C, then concentrated hydrochloric acid (12 N, ⁇ 1.5 mL) was slowly added until the pH of the solution was around 7. The mixture was then heated to 70-75 ° C and concentrated HCl (12 N, ⁇ 1 mL) was added dropwise to control the rate of carbon dioxide release until the pH was adjusted to about 3-4. The mixture was heated to 70-75 ° C and further reacted for 1 hour.
  • Example 8D To a mixture of Example 8D (500 mg, 2.05 mmol) and Boc 2 O (450 mg, 2.06 mmol) in methanol was added triethylamine (311 mg, 3.08 mmol), then the mixture was at 20-30 ° C Stir under 16 hours. The reaction mixture was concentrated, EtOAc EtOAc m. The crude intermediate was dissolved in ethanol (15 mL) and acetic acid (2 mL), then palladium hydroxide/carbon (0.1 g) was added, and the mixture was reacted under hydrogen (50 Psi) for 20 hours. The mixture was filtered, and the filtrate was evaporated to ethylamine
  • This example was prepared according to the method of Example 4A, replacing methyl t-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate with tert-butyl 3,9-diazaspiro[5.5 Tert-butyl undecyl-3-carboxylate.
  • the title compound was obtained as a yellow oil (yield: 62%).
  • This example was prepared according to the method of Example 7, (2-((5-chloro-2-(2,6-diazaspiro[3.4]oct-6-yl) Phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide was replaced by (2-((5-chloro-2-((2-methoxy-4-(3,9-) Diazaspiro[5.5]undecano-3-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide.
  • the title compound was obtained as a yellow solid, yield 57%.
  • Example 8C (100 mg, 0.18 mmol), sodium borohydride (114 mg, 0.54 mmol), acetic acid (21.6 mg, 0.36 mmol) and acetone (20.9 mg, 0.36 mmol) of tetrahydrofuran (5.0 mL) The mixture was stirred at 18 ° C for 16 hours. LCMS showed the reaction was complete. The solvent was evaporated to dryness crystals crystals crystals crystals LCMS (ESI) (10-80.): m/z:
  • This example was prepared according to the method of Example 4A, replacing the tert-butyl ester 2,7-diazaspiro[3.5]decane-2-carboxylic acid methyl ester with tert-butyl ester 3,9-diazaspiro[5.5 Tert-butane-3-carboxylic acid tert-butyl ester, and 4-fluoro-2-methoxy-1-nitrobenzene replaced with 2-(difluoromethoxy)-4-fluoro-1-nitrobenzene .
  • the title compound was obtained as a yellow oil, yield 77%.
  • This example was prepared according to the method of Example 7, (2-((5-chloro-2-(2,6-diazaspiro[3.4]oct-6-yl) Phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide was replaced by (2-((5-chloro-2-((2-fluorodimethoxy))-4-) 3,9-Diazaspiro[5.5]undecano-3-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide.
  • the title compound was obtained as a colorless oil, yield 22%.
  • Example 8A A mixed solution of Example 8A (0.7 g, 1.73 mmol) in trifluoroacetic acid (4 mL) and dichloromethane (4 mL) was stirred at 16 ° C for one hour. LCMS showed the reaction was completed. Aqueous sodium carbonate (50 mL) was added to the mixture and extracted with dichloromethane (50 mL ⁇ 2). The organic layer was dried with EtOAcjjjjjjjj LCMS (ESI) (0-60): m/z:
  • This example was prepared according to the method of Example 1M, replacing 2-methoxy-5-(6-methyl-6-azaspiro[3.4]oct-2-yl)aniline with 2-methoxy-4. -(9-Methyl-3,9-diazaspiro[5.5]undec-3-yl)aniline, and (2-((2,5-dichloropyrimidin-4-yl)amino)benzene
  • the dimethyl phosphine oxide is replaced by (2-((2,5-dichloropyrimidin-4-yl)amino)phenyl) dimethyl sulfide.
  • the title compound was obtained as a brown solid, yield 36%.
  • Example 13A 400.00 mg, 1.49 mmol
  • EtOAc 10 mL
  • TLC TLC showed the reaction was complete.
  • the mixture was concentrated under reduced pressure to give crystallite crystallite
  • Example 13B (251.00 mg, 1.49 mmol) and 4-fluoro-2-methoxy-1-nitrobenzene (305.98 mg, 1.79 mmol) in dimethyl sulfoxide (10 mL). Potassium carbonate (617.80 mg, 4.47 mmol) was added to the solution, and the mixture was stirred at 90 ° C for 12 hours. TLC showed the reaction was complete. The mixture was poured into water (25 mL) and extracted with dichloromethane (50 mL ⁇ 2).
  • Example 13C 310 mg, 0.97 mmol
  • MeOH 10 mL
  • TLC TLC showed complete consumption of the starting material.
  • the reaction mixture was filtered and evaporated to crystal crystal crystal crystal crystal crystal crystal crystal
  • Example 1M This example was prepared according to the method of Example 1M, replacing 2-methoxy-5-(6-methyl-6-azaspiro[3.4]oct-2-yl)aniline with 7-(4-amino- 3-methoxyphenyl)-N,N-dimethyl-7-azaspiro[3.5]decane-2-amine.
  • the title compound was obtained as a white solid in 16% yield.
  • This example was prepared according to the method of Example 13A, replacing ethyl t-butyl 2-oxo-7-azaspiro[3.5]decane-7-carboxylate with tert-butyl-9-oxo-3- Azaspiro[5.5]undecane-3-carboxylic acid methyl ester.
  • the title compound was obtained as a yellow oil (yield: 79.37%).
  • This example was prepared according to the method of Example 13B, replacing ethyl t-butyl 2-(dimethylamino)-7-azaspiro[3.5]decane-7-carboxylate with tert-butyl 10-( Methylamino)-3-azaspiro[5.5]undecane-3-carboxylic acid methyl ester.
  • the title compound was obtained as a yellow oil (yield).
  • Example 16B (50 mg, 0.08 mmol) was dissolved in TFA (3.0 mL). The solution was taken up in EtOAc (EtOAc)EtOAc.
  • Example 1K 300 mg, 1.77 mmol in DMF (5.
  • ⁇ RTI ID 0.0> ⁇ /RTI> ⁇ RTIgt; ⁇ /RTI> ⁇ RTIgt; ⁇ /RTI> ⁇ RTIgt; (2-(Trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (677 mg, 2.13 mmol)
  • the mixture was diluted with EtOAc (20 mL) andEtOAc.
  • the organic layer was dried and concentrated.
  • the crude product was purified by EtOAcjjjjjjjjj LCMS (ESI) (5-95AB): m/z: 4521.
  • the reaction suspension was diluted with ethyl acetate (20 mL) and filtered. The organic layer was washed with water (50 mL) and dried and evaporated.
  • the crude product was purified by EtOAcjjjjjjjjj LCMS (ESI) (5-95AB): m/z:
  • Example 1L to embodiment 100 mg, 0.32 mmol
  • K 2 CO 3 87.44 mg, 0.63 mmol
  • MeI 0.1mL, 1.58 mmol
  • EtOAc EtOAc
  • Example 12E (60 mg, 0.2 mmol), (2-((2,5-dichloropyrimidin-4-yl)oxy)phenyl)dimethyloxyphosphine (66 mg, 0.2 mmol), Xantphos (12 mg, 0.02 mmol), a mixture of palladium acetate (5 mg, 0.02 mmol), cesium carbonate (205 mg, 0.6 mmol) in dioxane (10 mL), heated to 100 ° C under N 2 and Stir for 12 hours. LCMS showed the reaction was completed. The reaction mixture was concentrated and purified EtOAcjjjjjjj LCMS (ESI) (5-95AB): m/z: 570.2 [M+1].
  • This example was prepared according to the method of Example 4A, replacing the methyl ester of tert-butyl ester 2,7-diazaspiro[3.5]nonane-2-carboxylate with 3,9-diazaspiro[5.5] eleven Tert-butyl-3-carboxylic acid, and 4-fluoro-2-methoxy-1-nitrobenzene were replaced by 1-chloro-2-fluoro-4-methoxy-5-nitrobenzene.
  • the title compound was obtained as a yellow oil.
  • This example was prepared according to the method of Example 7, (2-((5-chloro-2-(2,6-diazaspiro[3.4]oct-6-yl) Phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide was replaced by (2-((5-chloro-2-((5-chloro-2-methoxy-4-) 3,9-Diazaspiro[5.5]undecano-3-yl)phenyl)amino)pyridin-4-yl)amino)phenyl)dimethylphosphine oxide.
  • the title compound was obtained as a white solid in 40% yield.
  • Example 23B (70 mg, 0.42 mmol), 2-chloro-4-methoxy-5-nitropyridine (94 mg, 0.5 mmol), Pd 2 (dba) 3 (38)
  • a solution of Xantphos (24 mg, 0.04 mmol) and cesium carbonate (271 mg, 0.83 mmol) in dioxane (5 mL) was stirred at 90 ° C for 12 hours.
  • LCMS showed the reaction was complete.
  • the reaction mixture was diluted with water (40 mL) and evaporated with dichloromethane. The organic layer was washed with brine (30 mL EtOAc) The obtained residue was purified to mjjjjjjjjj
  • This example was prepared according to the method of Example 13D, 7-(3-methoxy-4-nitrophenyl)-N,N-dimethyl-7-azaspiro[3.5]decane-2-
  • the amine was replaced by 3-(4-methoxy-5-nitropyridin-2-yl)-9-methyl-3,9-diazaspiro[5.5]undecane.
  • the title compound was obtained as a brown solid in 88% yield.
  • This example was prepared according to the method of Example 1M, replacing 2-methoxy-5-(6-methyl-6-azaspiro[3.4]oct-2-yl)aniline with 2-methoxy-4. -(9-Methyl-3,9-diazaspiro[5.5]undec-3-yl)aniline, and (2-((2,5-dichloropyrimidin-4-yl)amino)benzene
  • the dimethylphosphine oxide is replaced by (2-((2,5-dichloropyrimidin-4-yl)amino)-3-fluorophenyl)dimethylphosphine oxide.
  • the title compound was obtained as a brown solid in 17% yield.
  • But-3-en-2-one (0.658 g, 9.39 mmol) was added to a solution of Example 27A (2 g, 9.39 mmol) in THF (100 mL).
  • the reaction mixture was cooled to -5 ° C, and a solution of potassium hydroxide-ethanol (3 mol / liter, 1.57 mL, 4.7 mmol) was added dropwise to the reaction mixture over 5 minutes.
  • the reaction mixture was warmed to 15 ° C and stirred for 16 hours. Petroleum ether (10 mL) was added to the reaction mixture and the mixture was washed with brine (100 mL). The organic layer was concentrated to give EtOAc (EtOAc:EtOAc. .
  • Example 27B To a solution of Example 27B (5.00 g, 18.84 mmol) in MeOH (100 mL). The suspension was evacuated and replaced several times with hydrogen. The reaction mixture was maintained at a temperature of 10-25 ° C under hydrogen (18 psi) and stirred for 5 hours. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by m ⁇ jjjjjjj 1 H NMR (400MHz, CDCl 3 ): ⁇ , 3.47-3.44 (m, 4H), 2.38-2.35 (m, 4H), 1.81-1.77 (m, 4H), 1.58-1.56 (m, 4H), 1.49 ( s, 9H).
  • Example 27C To a solution of Example 27C (2 g, 7.5 mmol) in tetrahydrofuran (20 mL) was added dropwise lithium diisopropylamide (2.5 M, 1.22 mL, 9 mmol). After that, the reaction mixture was stirred for 2 hours. Then, 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (2.67 g, 7.48 mmol, dissolved in 5 mL of THF) was added dropwise to the reaction. The solution was stirred at -78 ° C for 1.5 hours. The mixture was warmed to 10 <0>C and stirred for 2.5 h.
  • Example embodiments 27F (187 mg, 0.75 mmol) was added a solution of Pd (dppf) Cl 2 (15 mg, 0.075 mmol) and Potassium carbonate (204 mg, 1.5 mmol) was stirred at 110 ° C for 16 h. The reaction mixture was filtered through EtOAc EtOAc (EtOAc)EtOAc. Yellow oil. LCMS (ESI) (5-95AB): m.
  • This example was prepared according to the method of Example 7, (2-((5-chloro-2-(2,6-diazaspiro[3.4]oct-6-yl) Phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide was replaced by (2-((5-chloro-2-((2-methoxy-4-(3-aza)) Spiro[5.5]undecyl-9-yl)phenyl)amino)pyridin-4-yl)amino)phenyl)dimethylphosphine oxide.
  • the title compound was obtained as a yellow solid, yield 27%.
  • Example 11A This example was prepared according to the procedure of Example 11A, which was taken from 4-fluoro-2-methoxy-1-nitrobenzene to 4-chloro-2-methoxy-1-nitrobenzene. The title compound was obtained as a yellow oil.
  • Example 27D To a mixture of Example 27D (948 mg, 2.37 mmol), Example 28C (500 mg, 1.58 mmol) in dioxane (4 mL) and water (2 mL) Cl 2 (116 mg, 0.16 mmol) and sodium carbonate (503 mg, 4.75 mmol), and the mixture was stirred at 110 ° C for 16 hours. The reaction mixture was filtered through EtOAc EtOAc (EtOAc)EtOAc. Yellow solid.
  • This example was prepared according to the method of Example 1M, replacing 2-methoxy-5-(6-methyl-6-azaspiro[3.4]oct-2-yl)aniline with 9-(4-amino- Tert-butyl 3-(difluoromethoxy)phenyl)-3-azaspiro[5.5]undecane-3-carboxylate.
  • the title compound was obtained as a brown oil (37.2%).
  • 2,2,2-trichloroacetyl chloride was added dropwise to a mixture of Example 29A (6.00 g, 25.94 mmol) and zinc-copper (30.10 g, 233.46 mmol) in tetrahydrofuran (300 mL) at 15-20 °C (23.58 g, 129.70 mmol), after 0.5 hour dropwise, the reaction mixture was stirred at 20-30 ° C for 16 hours. The reaction solution was slowly quenched with ice and then extracted with ethyl acetate (250 mL ⁇ 3). The combined organic phases were washed with EtOAc EtOAc m.
  • Triethylsilicohydrogen (221.24 mg, 1.90 mmol) was added to a mixture of methylene chloride (10 mL). The reaction mixture was stirred at 18 ° C for 1 hour. The reaction mixture was poured into water (20 mL) and stirred for 20 min. The aqueous phase was extracted with ethyl acetate (30 mL ⁇ 3). The combined organic phases were washed with EtOAc EtOAc m. The residue was purified by EtOAc EtOAcjjjjjjj LCMS (ESI) (5-95AB): m.
  • Nitric acid (268.28 mg, 4.17 mmol) was added in one portion to a mixture of EtOAc (4 mL). The reaction mixture was stirred at 18 ° C for 2 hours. The reaction mixture was poured into ice water (20 mL) and stirred for 20 min. The aqueous phase was extracted with ethyl acetate (40 mL ⁇ 3). The combined organic phases were washed with brine (20 mL). Dry over anhydrous sodium sulfate, filtered and concentrated in vacuo.
  • Example 29F To a mixture of Example 29F (148.00 mg, 345.43 micromoles) in tetrahydrofuran (10 mL) was added palladium/carbon (40 mg, 10%). The reaction mixture was evacuated and replaced with H 2, the reaction mixture was stirred under hydrogen (16 psi) in 10-25 °C 3 hours. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by mjjjjlililililililililili LCMS (ESI) (5-95AB): m/z: 265.2 [M+1].
  • This example was prepared according to the method of Example 7, (2-((5-chloro-2-(2,6-diazaspiro[3.4]oct-6-yl) Phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide was replaced by (2-((5-chloro-2-(4-(2-fluoro-2-methoxy-4-) 7-Azaspiro[3.5]decane-2-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide.
  • the title compound was obtained as a yellow solid.
  • ALK wild type, ALK C1156Y and ALK L1196M were purchased from Carna Biosciences (Japan) and EGFR T790M/L858R was purchased from Life technology (Madison, WI).
  • HTRF kit purchased from Cis-Bio International containing Eu-labeled TK1 antibody, XL665 and biotin-tagged TK1 polypeptide substrate.
  • test compound was diluted 3-fold to obtain a final concentration of 11 doses from 1 uM to 0.017 nM.
  • 10 ul wild type ALK enzyme reaction mixture system 0.5 nM wild type ALK, 1 uM biotin-TK1 peptide, 30 uM ATP.
  • Reaction buffer 50 mM Hepes (pH 7.5), 10 mM MgCl2, 0.01 mM NaV3VO4.
  • the reaction plate was a white Proxiplate 384-Plus plate (PerkinElmer), and reacted at room temperature for 90 minutes.
  • 10 ul ALK C1156Y enzyme reaction mixture system 0.15 nM ALK C1156Y, 1 uM biotin-TK1 peptide, 30 uM ATP.
  • Reaction buffer 50 mM Hepes (pH 7.5), 10 mM MgCl2, 0.01 mM NaV3VO4.
  • the reaction plate was a white Proxiplate 384-Plus plate (PerkinElmer). The reaction was carried out for 60 minutes at room temperature.
  • reaction mixture system 0.15 nM ALK L1196M, 1 uM biotin-TK1 peptide, 30 uM ATP.
  • Reaction buffer 50 mM Hepes (pH 7.5), 10 mM MgCl 2 , 0.01 mM NaV 3 VO 4 .
  • the reaction plate was a white Proxiplate 384-Plus plate (PerkinElmer), and reacted at room temperature for 60 minutes.
  • 10 ul EGFR T790M/L858R enzyme reaction mixture system 0.08 nM EGFR T790M/L858R, 1 uM biotin-TK1 peptide, 20 uM ATP.
  • Reaction buffer 50 mM Hepes (pH 7.5), 10 mM MgCl2, 0.01 mM NaV3VO4.
  • the reaction plate was a white Proxiplate 384-Plus plate (PerkinElmer). The reaction was carried out for 60 minutes at room temperature.
  • Detection reaction 10 ul of detection reagent was added to the reaction plate, the final concentration of the Antibody was 2 nM, and the XL665 was 62.5 nM. Incubate for 60 minutes at room temperature. Envision reads the board.
  • the reading was converted to inhibition rate (%) by the following formula (Min-Ratio) / (Max-Min) * 100%.
  • 4 parameter curve fitting (Model 205in XLFIT5, iDBS) measured IC50 data.
  • RPMI 1640 fetal calf serum, penicillin/streptomycin solution, were purchased from Life Technology (Madison, WI).
  • Cell Titer-Glo luminescent cell viability reagents were purchased from Promega (Madison, WI).
  • Karpas299cell line is available from the European Collection of Cell Cultures (ECACC). Plate reading instrument: Envision (PerkinElmer).
  • 384-well plates 2500 Karpas-299 cells per well, 45 ul volume. Incubate overnight at 37 ° C in a CO 2 incubator. The test compound was diluted 3-fold to obtain 10 dose concentrations from 2.5 mM to 0.127 uM, two duplicate wells. The intermediate plate was added with 49 ul of medium per well. Transfer 1 ul of the compound from the gradient-diluted compound plate to the intermediate plate and mix well. Transfer 5 ul of liquid from the middle plate to the cell plate. The cells were further cultured for 72 hours in a CO2 incubator. After 72 hours, 25 ul of detection reagent was added. Incubate for 10 minutes at room temperature and Envision reads the plate. data analysis
  • the reading is converted to inhibition rate (%) by the following formula (Max-Sample) / (Max-Min) * 100% .
  • 4 parameter curve fitting (Model 205in XLFIT5, iDBS) measured IC 50 data.
  • the ALK enzyme of the compounds of the invention inhibits the IC50
  • the ALK L1196M enzyme inhibits the IC50
  • the ALK C1156Y enzyme inhibits the IC50
  • the EGFR T790M/L858R enzyme inhibits the IC50
  • the ALK IC50 of Karpas-299 cells is shown in the table below.
  • Compounds with IC 50 between 1-100 nM are indicated by +++; compounds with IC 50 between 101-1000 nm are represented by ++, and compounds with IC50 greater than 1000 nm are indicated by +.
  • the following in vivo pharmacodynamic data indicate that the compounds of the present invention are in a wild-type LU-01-0015 lung cancer patient-derived xenograft (PDX) model (BALB/c nude mice) and LU-01-0319 Crizotinib resistance model (BALB/c naked) Both models exhibited unexpected anti-tumor activity and reduced tumor volume compared to the reference compound AP26113.
  • PDX lung cancer patient-derived xenograft
  • BALB/c naked Both models exhibited unexpected anti-tumor activity and reduced tumor volume compared to the reference compound AP26113.
  • the tumor volume was reduced from approximately 277 mm 3 to 30- at the beginning. 45mm 3 , while AP26133 is only reduced to 119mm 3 .
  • mice Female, 6-8 weeks, weighing approximately 18-22 grams, maintained the mice in a special pathogen free environment and in a single ventilated cage (5 mice per cage). All cages, bedding and water are disinfected prior to use. All animals are free to access a standard certified commercial laboratory diet. A total of 80 mice purchased from Shanghai BK Laboratory Animal Co., LTD were used for the study. Tumor tissue (20-30 mm 3 ) was implanted subcutaneously into the right flank of each mouse for tumor growth. The experiment was started when the average tumor volume reached approximately 250-300 cubic millimeters. The test compound was orally administered daily, 25 mg/kg.
  • the anti-tumor efficacy is determined by dividing the average tumor-increased volume of the treated animals by the average tumor-increased volume of the untreated animals.
  • the LU-01-0319 xenograft model originally obtained from a surgically removed clinical sample, was implanted into nude mice, defined as batch P0 (LU-01-0319-P0).
  • the next batch of tumor implants from P0 was defined as batch P1 (LU-01-0319-P1).
  • FP3 was re-acquired from P2 and the next batch was defined as FP4 from FP3 tumor implantation.
  • tumor-bearing mice were treated with Crizotinib at a tumor size of about 300 mm3.
  • a continuously increasing tumor was defined as the LU-01-0319 anti-tumor model.
  • mice Female, 6-8 weeks, weighing about 18-22 grams, a total of 75 mice were used for research, purchased from Shanghai BK Laboratory Animal Co., LTD.
  • Tumor sections (about 30 mm 3 ) of LU-01-0319R FP6 were implanted subcutaneously into the right wing of each mouse for tumor growth.
  • Mice with tumors were treated with Crizotinib (10/25/50/75 mg/kg) approximately 2-3 weeks after tumor implantation and tumor size reached approximately 300 mm3 .
  • the dose of Crizotinib can be appropriately changed depending on the tumor size.
  • Experimental studies were initiated when the average tumor volume reached approximately ⁇ 500 mm3 .
  • the test compound is administered orally once a day.
  • the anti-tumor efficacy is determined by dividing the average tumor-increased volume of the treated animals by the average tumor-increased volume of the untreated animals.
  • mice Female, 6-8 weeks, weighing approximately 18-22 grams, maintained the mice in a special pathogen free environment and in a single ventilated cage (5 mice per cage). All cages, bedding and water are disinfected prior to use. All animals are free to access a standard certified commercial laboratory diet. A total of 80 mice purchased from Shanghai BK Laboratory Animal Co., LTD were used for the study. Tumor tissue (20-30 mm 3 ) was implanted subcutaneously into the right flank of each mouse for tumor growth. The experiment was started when the average tumor volume reached approximately 250-300 cubic millimeters. The test compound was orally administered daily, 10 mg/kg.
  • the anti-tumor efficacy is determined by dividing the average tumor-increased volume of the treated animals by the average tumor-increased volume of the untreated animals.
  • the ALK inhibitor of the present invention can be used for treating various cancers including anaplastic large cell lymphoma, non-small cell lung cancer, diffuse large B-cell lymphoma, inflammatory myofibroblastic tumor, neuroblastoma, thyroid undifferentiated Cancer and rhabdomyosarcoma.
  • ALK inhibitors can be used alone or in combination with other chemotherapeutic agents.

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Abstract

本发明公开了一种螺环芳基磷氧化物或硫化物作为ALK抑制剂，具体公开了作为ALK抑制剂的式（I）所示化合物或其药学上可接受的盐。

Description

螺环芳基磷氧化物和芳基磷硫化物 发明领域
本发明涉及一种螺环芳基磷氧化物或硫化物作为ALK抑制剂。具体地讲，本发明涉及作为ALK抑制剂的式(I)的化合物或其药学上可接受的盐。
发明背景
蛋白激酶几乎在所有类型的细胞生物活动中起主导调控作用。它们包括增殖，凋亡，细胞骨架重排，分化，发育，免疫反应，神经系统功能和传导。此外，许多疾病和(或)机能紊乱与一种或多种激酶的活性失常，异常或失调相关。
间变性淋巴瘤激酶(ALK)是酪氨酸激酶(RTKs)受体蛋白家族的一部分。ALK基因提供了指令使得酪氨酸激酶受体的蛋白，通过一个信号转导的过程，把信号从细胞表面传输到细胞内。这一过程始于当所述细胞表面激酶受到刺激，然后激酶发生二聚化。二聚化后，激酶被磷酸基标记，这一过程称为磷酸化。这一过程使激酶激活。激活的激酶能够将磷酸基团转移到细胞内的另一个蛋白上，并且继续将磷酸化传递到下游一系列蛋白。这种信号传导途径对许多细胞过程非常重要，如细胞生长和分裂(增殖)或成熟(分化)。
虽然对间变性淋巴瘤激酶的特异功能还不清楚，但一般认为在神经细胞早期发展中，它能够帮助调节神经细胞的增殖。
间变性淋巴瘤激酶ALK基因的突变是蛋白质的基本单元-氨基酸发生变化。在一些神经母细胞瘤，以及一种由未成熟的神经细胞(成神经细胞)组成的肿瘤病人中，已经鉴定出至少16个突变的ALK基因。神经母细胞瘤和其他癌症发生是由于一些关键基因(这些关键基因控制着细胞的增殖及分化)发生基因突变，使细胞生长和分裂不可控制而形成肿瘤。
神经母细胞瘤最常见的突变是在位置1275上，精氨酸被谷氨酸取代(写为Arg1275Gln或R1275Q)。Arg1275Gln突变在遗传性和分散性的神经母细胞瘤上都有发现，并且也是在这两种情况下发现的唯一共同的ALK基因突变。
突变或过度表达的间变性淋巴瘤激酶不再需要从细胞外刺激而磷酸化。因此，激酶和下游信号传导通路是不间断接通的(组成性激活)。间变性淋巴瘤激酶的组成性活化可提高未成熟神经细胞的增殖，最后导致神经母细胞瘤。ALK基因在2号染色体上的重排，增加了患有其它癌症的风险。这些重排是体细胞的突变，也就是体细胞的突变存在人的一生中，并且只在细胞发生癌变时才显现出来。
一种类型的重排称为易位，是在2号染色体和其他染色体之间的交换遗传物质。在患有间变性大细胞淋巴瘤(ALCL)的人群中，已鉴定出至少15种涉及ALK基因的易位。间变性大细胞淋巴瘤是发生在称为T细胞的免疫细胞中，是一种罕见的癌症形式。ALCL中最常见的易位发生染色体2和5号染色体之间，称为t(2，5)。这种易位使ALK基因与NPM基因融合，并形成NPM-ALK的融合蛋白。此外，在炎性肌纤维母细胞瘤(IMT)中已经鉴定了至少七种ALK基因易位。IMT是一种罕见的癌症，其特征是实体瘤由炎性细胞和一种对伤口愈合非常重要的肌纤维母细胞的组成。大约有一半IMT的人群涉及到ALK基因易位。
另一种类型的重排称为反转，反转发生在当2号染色体断裂成两部分，所得片断的DNA被倒置， 并重新插入到染色体中。非小细胞肺癌是一种最常见肺癌，在一小部分患者中，是2号染色体发生了反转。这种反转使ALK基因与另一个称为EML4的基因融合，形成EML4-ALK融合蛋白。由这些重排的基因产生的融合蛋白具有间变性淋巴瘤激酶和伴侣蛋白的双重功能。
近期ALK一直是抗肿瘤研发热门靶标，Mesatros总结了在这一领域的进展情况(Expert Opin.Ther Patents 2014,24(4),1)。Crizotinib是FDA批准的第一个ALK抑制剂，用于治疗ALK阳性肺癌。尽管经Crizotinib的最初响应非常有效，但多数患者由于产生耐药性而在治疗的第一年发生复发。2014年4月29日，FDA批准Ceritinib用于治疗间变性淋巴瘤激酶(ALK)阳性的转移性非小细胞肺癌(NSCLC)，包括对crizotinib有效以及耐药的患者。还有一些化合物正在临床研究中用于治疗癌症，如alectinib，AP-26113等。一些杂环化合物也已披露，用于各种癌症的治疗。专利包括WO2014033136，WO2014025128，WO2014006554，WO2014002922，WO2013192512，WO2013177092，WO2013148857，WO2013138210，WO2012139499，WO2012140114。
然而，尽管有一半以上的NSCLC患者对Crizotinib疗效良好，耐药总是会随用药时间而产生，从而药物失去其有效性。虽然近几年来，国内外都在大力开发用于治疗非小细胞肺癌的ALK抑制剂，但其疗效并不令人满意。因此非常迫切需要开发新型的，更加有效安全的ALK抑制剂。
本发明涉及一系列新型的螺环芳基磷氧化物和硫化物，该系列化合物是ALK和其突变体的抑制剂，可用于治疗癌症和其他疾病。本发明的新颖螺环芳基磷氧化物和硫化物出乎意料对ALK，ALK突变体与EGFR的突变酶展现出优于AP26113的抑制活性；并且这些化合物在ALK阳性病人中获得的NSCLC细胞系PDX模型及Crizotinib耐药PDX模型上也都展现出了比参考化合物AP26113更好的体内药效。因此，这些化合物对ALK酶异常所引起的疾病，可能会提供更加有效的治疗。
发明内容
本发明的目的在于提供式(I)所示化合物或其药学上可接受的盐，
Figure PCTCN2015082605-appb-000001
其中，
T1选自N或C(R01)；
T2选自-N(R01)-、O、S(＝O)2或-CH(NR01R02)-；
R01或R02分别独立地选自H、或任选被1、2或3个卤素、羟基和/或氰基所取代的：C1-6烷基、C1-6杂 烷基、C3-6环烷基-(CH2)0-3-和C3-6杂环烃基-(CH2)0-3-，其中所述“杂”代表1、2、或3个选自O、S、N、S(＝O)2和/或S(＝O)；
任选地，T2上的R01和R02相互连接到同一个N上形成1个3～6元环，所述的环上含有1、2或3个杂原子，所述杂原子选自O、S和N；
D1～D4分别独立地选自-(CR1R2)1-3-、O、S、C(＝O)、S(＝O)2和S(＝O)；
D选自-N(R01)-、-O-和-S-；
W选自＝O、＝S、＝N(CN)和＝N(OMe)；
R3选自R03、OR03和SR03；
R03选自C1-4烷基、C1-4卤代烷基和C3-5环烷基-(CH2)0-3-；
Z选自N和C(R4)；
X1选自C(Rx1)和N；
X2选自C(Rx2)和N；
Rx1、Rx2、R1、R2和R4分别独立地选自H、F、Cl、Br、I、CN、OH、SH、NH2、或任选被1、2或3个卤素、羟基和/或氰基所取代的：C1-6烷基、C1-6杂烷基、C3-6环烃基-(CH2)0-3-和C3-6杂环烃基-(CH2)0-3-，其中所述“杂”代表1、2、或3个杂原子，所述杂原子选自O、S和N；
任选地，Rx1和Rx2相互连接到同一个原子上形成1个5～6元环，所述的环上含有1、2或3个杂原子，所述杂原子选自O、S和N；
Rp1和Rp2分别独立地选自H、C1-4烷基和C1-4卤代烷基；
任选地，Rp1和Rp2相互连接到同一个P上形成1个5～6元环，所述的环上含有1、2或3个杂原子，所述杂原子选自O、S、N和P；和
任选地，Z和
Figure PCTCN2015082605-appb-000002
的位置可以互换。
本发明的一个方案中，上述R01和R02分别独立地选自H、CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3、-CH2CH2F、-CH2CH2S(＝O)2CH3、-CH2CH2CN、
Figure PCTCN2015082605-appb-000003
Figure PCTCN2015082605-appb-000004
-CH2CH(OH)(CH3)2、-CH2CH(F)(CH3)2和-CH2CH2F。
本发明的一个方案中，上述R03选自CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3、-CH2CH2F和
Figure PCTCN2015082605-appb-000005
本发明的一个方案中，上述Rp1和Rp2分别独立地选自H、CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3和-CH2CH2F。
本发明的一个方案中，上述Rx1、Rx2、R1、R2和R4分别独立地选自H、F、Cl、Br、I、CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3、-CH2CH2F、-CH2CH2S(＝O)2CH3、-CH2CH2CN、
Figure PCTCN2015082605-appb-000006
-CH2CH(OH)(CH3)2,-CH2CH(F)(CH3)2和-CH2CH2F。
本发明的一个方案中，上述T2上的NR01R02选自NHCH3、N(CH3)2、N(CH2CH3)2、
Figure PCTCN2015082605-appb-000007
Figure PCTCN2015082605-appb-000008
本发明的一个方案中，上述D、D1-4或T2分别独立地选自–NH-、-NMe-和-O-；D1-4和T2还可以选自-CH(NCH3CH3)-。
本发明的一个方案中，上述螺环结构单元
Figure PCTCN2015082605-appb-000009
选自
Figure PCTCN2015082605-appb-000010
Figure PCTCN2015082605-appb-000011
本发明的一个方案中，上述化合物或其药学上可接受的盐，其具有式(Ⅱ)所示结构：
Figure PCTCN2015082605-appb-000012
其中各变量如上述定义。
本发明的一个方案中，上述化合物或其药学上可接受的盐，其具有式(Ⅲ)所示结构：
Figure PCTCN2015082605-appb-000013
其中各变量如上述定义。
本发明的一个方案中，上述化合物或其药学上可接受的盐，其选自：
Figure PCTCN2015082605-appb-000014
Figure PCTCN2015082605-appb-000015
本发明的另一目的在于提供制备上述化合物的方法，其制备路线如方案A或C所示：
方案A
Figure PCTCN2015082605-appb-000016
其中A5的制备方法如方案B所示：
方案B
Figure PCTCN2015082605-appb-000017
本发明的的另一目的在于提供上述化合物在制备治疗ALK和/或EGFR以及它们的突变引起的非小细胞肺癌及其它癌症、与ROS1、BRAF、c-MET、HER2、KRAS/MEK、PIK3CA、FDFR、DDR2和/或VEGFR等抑制剂联合治疗的癌症、以及与细胞毒素如多西他赛或卡铂等联合治疗的癌症的药物中的应用。
定义和说明：
C1-6选自于由C1，C2，C3，C4，C5和C6组成的基团，数字表示碳原子数的基团；C3-6选自于由C3，C4，C5，和C6组成的基团。
C1-6烷基，C1-6杂烷基，C3-6环烷基，C3-6杂环烷基，C1-6烷基被C3-6环烷基或C3-6杂环烷基取代，和C1-6杂烷基被C3-6环烷基或C3-6杂环烷基取代，包括但不限于：甲基，乙基，正丙基，异丙基，-CH2C(CH3)(CH3)(OH)，环丙基，环丁基，丙基亚甲基，环丙基酰基，苄氧基，环丙基烯基，三氟甲基，氨基甲基，羟甲基，甲氧基，甲基酰基，甲氧酰基，甲基磺酰基，甲基亚磺酰基，乙氧基，乙酰基，乙基磺酰基，乙氧酰基，二甲基氨基，二乙基氨基，二甲基氨基，和二乙基氨基；N(CH3)2,NH(CH3),-CH2CF3,-CH2CH2CF3,-CH2CH2F,-CH2CH2S(＝O)2CH3,-CH2CH2CN,
Figure PCTCN2015082605-appb-000018
Figure PCTCN2015082605-appb-000019
-CH2CH(OH)(CH3)2,-CH2CH(F)(CH3)2,-CH2CH2F,-CH2CF3,-CH2CH2CF3,-CH2CH2NH2,-CH2CH2OH,-CH2CH2OCH3,-CH2CH2CH2OCH3,-CH2CH2N(CH3)2,-S(＝O)2CH3,-CH2CH2S(＝O)2CH3,
Figure PCTCN2015082605-appb-000020
Figure PCTCN2015082605-appb-000021
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏 性反应或其它问题或并发症，与合理的利益/风险比相称。
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物的中性形式接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机氨或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物的中性形式接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，所述无机酸包括例如盐酸、氢溴酸、硝酸、碳酸，碳酸氢根，磷酸、磷酸一氢根、磷酸二氢根、硫酸、硫酸氢根、氢碘酸、亚磷酸等；以及有机酸盐，所述有机酸包括如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸和甲磺酸等类似的酸；还包括氨基酸(如精氨酸等)的盐，以及如葡糖醛酸等有机酸的盐(参见Berge et al.,"Pharmaceutical Salts",Journal of Pharmaceutical Science 66:1-19(1977))。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。
优选地，以常规方式使盐与碱或酸接触，再分离母体化合物，由此再生化合物的中性形式。化合物的母体形式与其各种盐的形式的不同之处在于某些物理性质，例如在极性溶剂中的溶解度不同。
本文所用的“药学上可接受的盐”属于本发明化合物的衍生物，其中，通过与酸成盐或与碱成盐的方式修饰所述母体化合物。药学上可接受的盐的实例包括但不限于：碱基比如胺的无机酸或有机酸盐、酸根比如羧酸的碱金属或有机盐等等。药学上可接受的盐包括常规的无毒性的盐或母体化合物的季铵盐，例如无毒的无机酸或有机酸所形成的盐。常规的无毒性的盐包括但不限于那些衍生自无机酸和有机酸的盐，所述的无机酸或有机酸选自2-乙酰氧基苯甲酸、2-羟基乙磺酸、乙酸、抗坏血酸、苯磺酸、苯甲酸、碳酸氢根、碳酸、柠檬酸、依地酸、乙烷二磺酸、乙烷磺酸、富马酸、葡庚糖、葡糖酸、谷氨酸、乙醇酸、氢溴酸、盐酸、氢碘酸盐、羟基、羟萘、羟乙磺酸、乳酸、乳糖、十二烷基磺酸、马来酸、苹果酸、扁桃酸、甲烷磺酸、硝酸、草酸、双羟萘酸、泛酸、苯乙酸、磷酸、多聚半乳糖醛、丙酸、水杨酸、硬脂酸、亚乙酸、琥珀酸、氨基磺酸、对氨基苯磺酸、硫酸、单宁、酒石酸和对甲苯磺酸。
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。一般地，优选醚、乙酸乙酯、乙醇、异丙醇或乙腈等非水介质。
除了盐的形式，本发明所提供的化合物还存在前药形式。本文所描述的化合物的前药容易地在生理条件下发生化学变化从而转化成本发明的化合物。此外，前体药物可以在体内环境中通过化学或生化方法被转换到本发明的化合物。
本发明的某些化合物可以以非溶剂化形式或者溶剂化形式存在，包括水合物形式。一般而言，溶剂化形式与非溶剂化的形式相当，都包含在本发明的范围之内。本发明的某些化合物可以以多晶或无定形形式存在。
本发明的某些化合物可以具有不对称碳原子(光学中心)或双键。外消旋体、非对映异构体、几何异构体和单个的异构体都包括在本发明的范围之内。
本文中消旋体、ambiscalemic and scalemic或者对映体纯的化合物的图示法来自Maehr,J.Chem.Ed.1985,62:114-120。1985年，62：114-120。除非另有说明，用楔形键和虚线键表示一个立体中心的绝对构型。当本文所述化合物含有烯属双键或其它几何不对称中心，除非另有规定，它们包括E、Z几何异构体。同样地，所有的互变异构形式均包括在本发明的范围之内。
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。
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的分步结晶法或色谱法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。
本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚(3H)，碘-125(125I)或C-14(14C)。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。
术语“药学上可接受的载体”是指能够递送本发明有效量活性物质、不干扰活性物质的生物活性并且对宿主或者患者无毒副作用的任何制剂或载体介质代表性的载体包括水、油、蔬菜和矿物质、膏基、洗剂基质、软膏基质等。这些基质包括悬浮剂、增粘剂、透皮促进剂等。它们的制剂为化妆品领域或局部药物领域的技术人员所周知。关于载体的其他信息，可以参考Remington:The Science and Practice of Pharmacy,21st Ed.,Lippincott,Williams&Wilkins(2005)，该文献的内容通过引用的方式并入本文。
术语“赋形剂”通常是指配制有效的药物组合物所需要载体、稀释剂和/或介质。
针对药物或药理学活性剂而言，术语“有效量”或“治疗有效量”是指无毒的但能达到预期效果的药物或药剂的足够用量。对于本发明中的口服剂型，组合物中一种活性物质的“有效量”是指与该组合物中另一种活性物质联用时为了达到预期效果所需要的用量。有效量的确定因人而异，取决于受体的年龄和一般情况，也取决于具体的活性物质，个案中合适的有效量可以由本领域技术人员根据常规试验确定。
术语“活性成分”、“治疗剂”、“活性物质”或“活性剂”是指一种化学实体，它可以有效地治疗目标紊乱、疾病或病症。
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，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。当取代基为酮基(即＝O)时，意味着两个氢原子被取代。酮取代不会发生在芳香基上。术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。
当一个取代基的键可以交叉连接到一个环上的两个原子时，这种取代基可以与这个环上的任意原子相键合。当所列举的取代基中没有指明其通过哪一个原子连接到化学结构通式中包括但未具体提及的化合物时，这种取代基可以通过其任何原子相键合。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。
烷基和杂烷基原子团(包括通常被称为亚烷基、链烯基、亚杂烷基、杂烯基、炔基、环烷基、杂环烷基、环烯基和杂环烯基的那些基团)的取代基一般被称为“烷基取代基”，它们可以选自但不限于下列基团中的一个或多个：-R’、-OR’、＝O、＝NR’、＝N-OR’、-NR’R”、-SR’、卤素、-SiR’R”R”’、OC(O)R’、-C(O)R’、-CO2R’、-CONR’R”、-OC(O)NR’R”、-NR”C(O)R’、NR’C(O)NR”R”’、-NR”C(O)2R’、-NR””’-C(NR’R”R”')＝NR””、NR””C(NR’R”)＝NR”'、-S(O)R’、-S(O)2R’、-S(O)2NR’R”、NR”SO2R’、-CN、–NO2、-N3、-CH(Ph)2和氟代(C1-C4)烷基,取代基的数目为0～(2m’+1),其中m’是这类原子团中碳原子的总数。R'、R”、R”'、R””和R””’各自独立地优选氢、被取代或未被取代的杂烷基、被取代或未被取代的芳基(例如被1～3个卤素取代芳基)、被取代或未被取代的烷基、烷氧基、硫代烷氧基基团或芳烷基。当本发明的化合物包括一个以上的R基团时，例如，每一个R基团是独立地加以选择的，如同当存在一个以上的R'、R”、R”'、R””和R””’基团时的每个这些基团。当R'和R”附着于同一个氮原子时，它们可与该氮原子结合形成5-，6-或7-元环。例如，-NR'R“意在包括但不仅限于1-吡咯烷基和4-吗啉基。根据上述关于取代基的讨论中，本领域技术人员可以理解，术语“烷基”意在包括碳原子键合于非氢基团所构成的基团，如卤代烷基(例如-CF3、-CH2CF3)和酰基(例如-C(O)CH3、-C(O)CF3、-C(O)CH2OCH3等)。与烷基原子团所述取代基相似，芳基和杂芳基取代基一般统称为“芳基取代基”，选自例如-R’、-OR’、-NR’R”、-SR’、-卤素,-SiR’R”R”’、OC(O)R’、-C(O)R’、-CO2R’、-CONR’R”、-OC(O)NR’R”、-NR”C(O)R’、NR’C(O)NR”R”’、-NR”C(O)2R’、-NR””’-C(NR’R”R”')＝NR””、NR””C(NR’R”)＝NR”'、-S(O)R’、-S(O)2R’、-S(O)2NR’R”、NR”SO2R’、-CN、–NO2、-N3、-CH(Ph)2、氟(C1-C4)烷氧基和氟(C1-C4)烷基等，取代基的数量为0到芳香环上开放化合价的总数之间；其中R’、R”、R”’、R””和R””’独立地优选自氢、被取代或未被取代的烷基、被取代或未被取代的杂烷基、被取代或未被取代的芳基和被取代或未被取代的杂芳基。当本发明的化合物包括一个以上的R基团时，例如，每个R基团是独立地加以选择的，如同当存在一个以上R’、R”、R”’、R””和R””’基团时的每个这些基团。
芳基或杂芳基环的相邻原子上的两个取代基可以任选地被通式为–T-C(O)-(CRR’)q-U-的取代基所取代，其中T和U独立地选自-NR-、-O-、CRR'-或单键，q是0到3的整数。作为替代选择，芳基或杂芳基环的相邻原子上的两个取代基可以任选地被通式为–A(CH2)r B-的取代基所取代，其中A和B独立的选自–CRR’-、-O-、-NR-、-S-、-S(O)-、S(O)2-、-S(O)2NR’-或单键，r是1～4的整数。任选地，由此形成的新环上的一个单键可以替换为双键。作为替代选择，芳基或杂芳基环的相邻原子上的两个取代基可以任选地被通式为–A(CH2)r B-的取代基所取代，其中s和d分别独立的选自0～3的整数，X是–O-、 -NR’、-S-、-S(O)-、-S(O)2-或–S(O)2NR’-。取代基R、R’、R”和R”’分别独立地优选自氢和被取代或未被取代的(C1-C6)烷基。
除非另有规定，术语“卤代素”或“卤素”本身或作为另一取代基的一部分表示氟、氯、溴或碘原子。此外，术语“卤代烷基”意在包括单卤代烷基和多卤代烷基。例如，术语“卤代(C1-C4)烷基”意在包括但不仅限于三氟甲基、2，2，2-三氟乙基、4-氯丁基和3-溴丙基等等。
卤代烷基的实例包括但不仅限于：三氟甲基、三氯甲基、五氟乙基，和五氯乙基。“烷氧基”代表通过氧桥连接的具有特定数目碳原子的上述烷基。C1-6烷氧基包括C1、C2、C3、C4、C5和C6的烷氧基。烷氧基的例子包括但不限于：甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、仲丁氧基、叔丁氧基、正戊氧基和S-戊氧基。“环烷基”包括饱和环基，如环丙基、环丁基或环戊基。3-7环烷基包括C3、C4、C5、C6和C7环烷基。“链烯基”包括直链或支链构型的烃链，其中该链上任何的稳定位点上存在一个或多个碳-碳双键，例如乙烯基和丙烯基。
术语“卤”或“卤素”是指氟、氯、溴和碘。
本文所用术语“杂原子”包括碳(C)和氢(H)以外的原子，例如包括氧(O)、氮(N)、硫(S)、硅(Si)、锗(Ge)、铝(Al)和硼(B)等。
除非另有说明，术语“杂”，“杂原子”或“杂自由基”(即自由基含杂原子)，包括除碳(C)和氢(H)外的原子，也包含上述杂原子的自由基。例如包括氧(O)、氮(N)、硫(S)、硅(Si)、锗(Ge)、铝(Al)和硼(B)等，还包括任意取代的-C(＝O)N(H)-、-N(H)-、-C(＝NH)-、-S(＝O)2N(H)-,or-S(＝O)N(H)-。
“环”表示被取代或未被取代的环烷基、被取代或未被取代的杂环烷基、被取代或未被取代的芳基或被取代或未被取代的杂芳基。所谓的环包括稠环。环上原子的数目通常被定义为环的元数，例如，“5～7元环”是指环绕排列5～7个原子。除非另有规定，该环任选地包含1～3个杂原子。因此，“5～7元环”包括例如苯基吡啶和哌啶基；另一方面，术语“5～7元杂环烷基环”包括吡啶基和哌啶基，但不包括苯基。术语“环”还包括含有至少一个环的环系，其中的每一个“环”均独立地符合上述定义。
术语“杂环”或“杂环基”意指稳定的单环或双环或双环杂环，它们可以是饱和的、部分不饱和的或不饱和的(芳族的)，它们包含碳原子和1、2、3或4个独立地选自N、O和S的环杂原子，其中上述任意杂环可以稠合到一个苯环上形成双环。氮和硫杂原子可任选被氧化(即NO和S(O)p)。氮原子可以是被取代的或未取代的(即N或NR，其中R是H或本文已经定义过的其他取代基)。该杂环可以附着到任何杂原子或碳原子的侧基上从而形成稳定的结构。如果产生的化合物是稳定的，本文所述的杂环可以发生碳位或氮位上的取代。杂环中的氮原子任选地被季铵化。一个优选方案是，当杂环中S及O原子的总数超过1时，这些杂原子彼此不相邻。另一个优选方案是，杂环中S及O原子的总数不超过1。如本文所用，术语“芳族杂环基团”或“杂芳基”意指稳定的5、6、7元单环或双环或7、8、9或10元双环杂环基的芳香环，它包含碳原子和1、2、3或4个独立地选自N、O和S的环杂原子。氮原子可以是被取代的或未取代的(即N或NR，其中R是H或本文已经定义过的其他取代基)。氮和硫杂原子可任选被氧化(即NO和S(O)p)。值得注意的是，芳香杂环上S和O原子的总数不超过1。桥环也包含在杂环的定义中。当一个或多个原子(即C、O、N或S)连接两个不相邻的碳原子或氮原子时形成桥环。优选的桥环包括但不限于：一个碳原子、两个碳原子、一个氮原子、两个氮原子和一个碳-氮基。值得 注意的是，一个桥总是将单环转换成三环。桥环中，环上的取代基也可以出现在桥上。
杂环化合物的实例包括但不限于：吖啶基、吖辛因基、苯并咪唑基、苯并呋喃基、苯并巯基呋喃基、苯并巯基苯基、苯并恶唑基、苯并恶唑啉基、苯并噻唑基、苯并三唑基、苯并四唑基、苯并异恶唑基、苯并异噻唑基、苯并咪唑啉基、咔唑基、4aH-咔唑基、咔啉基、苯并二氢吡喃基、色烯、噌啉基十氢喹啉基、2H,6H-1,5,2-二噻嗪基、二氢呋喃并[2,3-b]四氢呋喃基、呋喃基、呋咱基、咪唑烷基、咪唑啉基、咪唑基、1H-吲唑基、吲哚烯基、二氢吲哚基、中氮茚基、吲哚基、3H-吲哚基、isatino基、异苯并呋喃基、异吲哚基、异二氢吲哚基、异喹啉基、异噻唑基、异恶唑基、亚甲二氧基苯基、吗啉基、萘啶基，八氢异喹啉基、恶二唑基、1,2,3-恶二唑基、1,2,4-恶二唑基、1,2,5-恶二唑基、1,3,4-恶二唑基、恶唑烷基、恶唑基、羟吲哚基、嘧啶基、菲啶基、菲咯啉基、吩嗪、吩噻嗪、苯并黄嘌呤基、酚恶嗪基、酞嗪基、哌嗪基、哌啶基、哌啶酮基、4-哌啶酮基、胡椒基、蝶啶基、嘌呤基、吡喃基、吡嗪基、吡唑烷基、吡唑啉基、吡唑基、哒嗪基、吡啶并恶唑、吡啶并咪唑、吡啶并噻唑、吡啶基、吡咯烷基、吡咯啉基、2H-吡咯基、吡咯基、喹唑啉基、喹啉基、4H-喹嗪基、喹喔啉基、奎宁环基、四氢呋喃基、四氢异喹啉基、四氢喹啉基、四唑基，6H-1,2,5-噻二嗪基、1,2,3-噻二唑基、1,2,4-噻二唑基、1,2,5-噻二唑基、1,3,4-噻二唑基、噻蒽基、噻唑基、异噻唑基噻吩基、噻吩并恶唑基、噻吩并噻唑基、噻吩并咪唑基、噻吩基、三嗪基、1,2,3-三唑基、1,2,4-三唑基、1,2,5-三唑基、1,3,4-三唑基和呫吨基。还包括稠环和螺环化合物。
除非另有规定，术语“烃基”或者其下位概念(比如烷基、烯基、炔基、苯基等等)本身或者作为另一取代基的一部分表示直链的、支链的或环状的烃原子团或其组合，可以是完全饱和的、单元或多元不饱和的，可以是单取代、二取代或多取代的，可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)，可以包括二价或多价原子团，具有指定数量的碳原子(如C1-C10表示1至10个碳)。“烃基”包括但不限于脂肪烃基和芳香烃基，所述脂肪烃基包括链状和环状，具体包括但不限于烷基、烯基、炔基，所述芳香烃基包括但不限于6-12元的芳香烃基，例如苯、萘等。在一些实施例中，术语“烷基”表示直链的或支链的原子团或它们的组合，可以是完全饱和的、单元或多元不饱和的，可以包括二价和多价原子团。饱和烃原子团的实例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、叔丁基、异丁基、仲丁基、异丁基、环己基、(环己基)甲基、环丙基甲基，以及正戊基、正己基、正庚基、正辛基等原子团的同系物或异构体。不饱和烷基具有一个或多个双键或三键，其实例包括但不限于乙烯基、2-丙烯基、丁烯基、巴豆基、2-异戊烯基、2-(丁二烯基)、2，4-戊二烯基、3-(1，4-戊二烯基)、乙炔基、1-和3-丙炔基，3-丁炔基，以及更高级的同系物和异构体。
除非另有规定，术语“杂烃基”或者其下位概念(比如杂烷基、杂烯基、杂炔基、杂芳基等等)本身或者与另一术语联合表示稳定的直链的、支链的或环状的烃原子团或其组合，有一定数目的碳原子和至少一个杂原子组成。在一些实施例中，术语“杂烷基”本身或者与另一术语联合表示稳定的直链的、支链的烃原子团或其组合物，有一定数目的碳原子和至少一个杂原子组成。在一个典型实施例中，杂原子选自B、O、N和S，其中氮和硫原子任选地被氧化，氮杂原子任选地被季铵化。杂原子B、O、N和S可以位于杂烃基的任何内部位置(除该烃基基附着于分子其余部分的位置之外)。实例包括但不限于-CH2-CH2-O-CH3、-CH2-CH2-NH-CH3、-CH2-CH2-N(CH3)-CH3、-CH2-S-CH2-CH3、-CH2-CH2、-S(O)-CH3、 -CH2-CH2-S(O)2-CH3、-CH＝CH-O-CH3、-CH2-CH＝N-OCH3和–CH＝CH-N(CH3)-CH3。至多两个杂原子可以是连续的，例如-CH2-NH-OCH3。
术语“烷氧基”、“烷氨基”和“烷硫基”(或硫代烷氧基)属于惯用表达，是指分别通过一个氧原子、氨基或硫原子连接到分子的其余部分的那些烷基基团。
除非另有规定，术语“环烃基”、“杂环烃基”、“环烃杂基”或者其下位概念(比如芳基、杂芳基、芳杂基、环烷基、杂环烷基、环烷杂基、环烯基、杂环烯基、环烯杂基、环炔基、杂环炔基、环炔杂基等等)本身或与其他术语联合分别表示环化的“烃基”、“杂烃基”或“烃杂基”。此外，就杂烃基或杂环烃基(比如杂烷基、杂环烷基)而言，杂原子可以占据该杂环附着于分子其余部分的位置。环烷基的实例包括但不限于环戊基、环己基、1-环己烯基、3-环己烯基、环庚基等。杂环基的非限制性实例包括1-(1,2,5,6-四氢吡啶基)、1-哌啶基、2-哌啶基，3-哌啶基、4-吗啉基、3-吗啉基、四氢呋喃-2-基、四氢呋喃吲哚-3-基、四氢噻吩-2-基、四氢噻吩-3-基，1-哌嗪基和2-哌嗪基。
除非另有规定，术语“芳基”表示多不饱和的芳族烃取代基，可以是单取代、二取代或多取代的，它可以是单环或多环(优选1至3个环)，它们稠合在一起或共价连接。术语“杂芳基”是指含有一至四个杂原子的芳基(或环)。在一个示范性实例中，杂原子选自B、N、O和S，其中氮和硫原子任选地被氧化，氮原子任选地被季铵化。杂芳基可通过杂原子连接到分子的其余部分。芳基或杂芳基的非限制性实施例包括苯基、1-萘基、2-萘基、4-联苯基、1-吡咯基、2-吡咯基、3-吡咯基、3-吡唑基、2-咪唑基、4-咪唑基、吡嗪基、2-恶唑基、4-恶唑基、2-苯基-4-恶唑基、5-恶唑基、3-异恶唑基、4-异恶唑基、5-异恶唑基、2-噻唑基、4-噻唑基、5-噻唑基、2-呋喃基、3-呋喃基、2-噻吩基、3-噻吩基、2-吡啶基、3-吡啶基、4-吡啶基、2-嘧啶基、4-嘧啶基、5-苯并噻唑基、嘌呤基、2-苯并咪唑基、5-吲哚基、1-异喹啉基、5-异喹啉基、2-喹喔啉基、5-喹喔啉基、3-喹啉基和6-喹啉基。上述任意一个芳基和杂芳基环系的取代基选自下文所述的可接受的取代基。
为简便起见，芳基在与其他术语联合使用时(例如芳氧基、芳硫基、芳烷基)包括如上定义的芳基和杂芳基环。因此，术语“芳烷基”意在包括芳基附着于烷基的那些原子团(例如苄基、苯乙基、吡啶基甲基等)，包括其中碳原子(如亚甲基)已经被例如氧原子代替的那些烷基，例如苯氧基甲基、2-吡啶氧甲基3-(1-萘氧基)丙基等。
术语“离去基团”是指可以被另一种官能团或原子通过取代反应(例如亲和取代反应)所取代的官能团或原子。例如，代表性的离去基团包括三氟甲磺酸酯；氯、溴、碘；磺酸酯基，如甲磺酸酯、甲苯磺酸酯、对溴苯磺酸酯、对甲苯磺酸酯等；酰氧基，如乙酰氧基、三氟乙酰氧基等等。
术语“保护基”包括但不限于“氨基保护基”、“羟基保护基”或“巯基保护基”。术语“氨基保护基”是指适合用于阻止氨基氮位上副反应的保护基团。代表性的氨基保护基包括但不限于：甲酰基；酰基，例如链烷酰基(如乙酰基、三氯乙酰基或三氟乙酰基)；烷氧基羰基，如叔丁氧基羰基(Boc)；芳基甲氧羰基，如苄氧羰基(Cbz)和9-芴甲氧羰基(Fmoc)；芳基甲基，如苄基(Bn)、三苯甲基(Tr)、1,1-二-(4'-甲氧基苯基)甲基；甲硅烷基，如三甲基甲硅烷基(TMS)和叔丁基二甲基甲硅烷基(TBS)等等。术语“羟基保护基”是指适合用于阻止羟基副反应的保护基。代表性羟基保护基包括但不限于：烷基，如甲基、乙基和叔丁基；酰基，例如链烷酰基(如乙酰基)；芳基甲基，如苄基(Bn)，对甲氧基苄基(PMB)、 9-芴基甲基(Fm)和二苯基甲基(二苯甲基，DPM)；甲硅烷基，如三甲基甲硅烷基(TMS)和叔丁基二甲基甲硅烷基(TBS)等等。
通式制备例：
在一些实施例中，式(I)的中的部分化合物可以按照方案A中所述的合成方法制备，其中T1代表N，T2代表NH。
方案A
Figure PCTCN2015082605-appb-000022
其中PG是氨基保护基，优选自BOC、Bn和Cbz。
膦酸酯(A1)与格氏试剂反应，例如与甲基溴化镁在溶剂四氢呋喃或乙醚中反应，得到氧化磷(A2)。偶合反应是在钯催化下在溶剂中如DMF，以及碱存在下如碳酸钾，得到芳基磷氧化合物A3；A3可以通过劳森试剂的作用被转化为磷硫化合物。芳基磷氧化物或硫化物与二氯嘧啶衍生物或其它二氯杂环化合物反应，得到化合物A4，它可以与芳基胺A7在酸性条件下偶联，例如在甲烷磺酸存在下，以叔丁醇为溶剂反应得到本发明化合物A8，或可以进一步反应，如还原胺化，得到式(I)的化合物。螺环胺基化合物A5可以从化学供应商处购买，例如Aldrich Chemical，或者可以通过以下的方案B来制备。螺环氨基化合物A5通过在溶剂(如DMF)中发生取代反应，得到化合物A6，通过进一步加氢还原得到芳基胺A7。
方案B
Figure PCTCN2015082605-appb-000023
方案B为螺环胺类化合物B5(A5)的一般合成方法，酮B1与氰基乙酸酯反应，得到螺环化合物B2，水解得到B3，然后B4，用还原剂如氢化铝锂，还原B4得到B5；B5可以与方案A中的芳基氯A4偶联得到A6。
方案C
Figure PCTCN2015082605-appb-000024
方案C是制备式(I)化合物的一般合成方法，其中T1是CH。C1化合物醛与甲基乙烯基酮，在碱如KOH作用下反应，得到酮C2，氢化得到C3，三氟甲磺酸酯化生成C4；然后与氨基芳基或硝基芳硼酸偶联得到C5，还原得到化合物C6；然后与化合物A4偶联，得到本发明化合物C7，可以进一步反应，如还原胺化，得到式(I)的化合物。
本发明现在进一步通过实施例描述。下面给出的实施例仅用于说明目的，而不是仅限于此发明的范 围。本发明的化合物可以用有机合成领域中许多已知的方法来制备。本发明的实施例可以使用下面描述的方法来合成，以及有机合成化学领域中已知的合成方法，或在其基础上通过改进的方法。优选的方法包括，但不限于以下描述方法。
本发明所使用的所有溶剂是市售的，无需进一步纯化即可使用。反应一般是在惰性氮气下、无水溶剂中进行的。质子核磁共振数据记录在Bruker Avance III 400(400MHz)分光仪上，化学位移以四甲基硅烷低场处的(ppm)表示。质谱是在安捷伦1200系列加6110(&1956A)上测定。LC/MS或Shimadzu MS包含一个DAD：SPD-M20A(LC)和Shimadzu Micromass 2020检测器。质谱仪配备有一个正或负模式下操作的电喷雾离子源(ESI)。
本发明采用下述缩略词：aq代表水；DCM代表二氯甲烷；PE代表石油醚；DMF代表N，N-二甲基甲酰胺；DMSO代表二甲基亚砜；EtOAc代表乙酸乙酯；EtOH代表乙醇；MeOH代表甲醇；Cbz代表苄氧羰基，一种胺保护基；Boc代表叔丁基氧羰基，一种胺保护基；HOAc代表乙酸；NaBH(OAc)3代表三乙酰氧基硼氢化钠；r.t代表室温；THF代表四氢呋喃；Boc2O代表二叔丁基二碳酸酯；TFA代表三氟乙酸；DIPEA代表二异丙基乙胺；Pd(dppf)Cl2代表[1,1'-双(二苯基膦基)二茂铁]二氯化钯(II)；POCl3代表三氯氧磷；NaH代表氢化钠；LAH代表氢化铝锂；Pd(OAc)2代表钯(II)乙酸盐；Pd2(dba)3代表三(二亚苄基丙酮)二钯；Pd(PPh3)4代表四(三苯基膦)钯；Et3SiH代表三乙基硅烷；PPh3代表三苯基膦；Xantphos代表4,5-双(二苯基膦基)-9,9-二甲基；MeSO3H代表甲磺酸；Xphos代表2-二环己基膦基-2'，4'，6'-三异丙基联苯；劳森试剂代表2,4-二(4-甲氧基苯基)-1,3-二硫杂-2,4-二磷烷-2,4-二硫化物；NBS代表N-溴代丁二酰亚胺；t-BuOK代表叔丁醇钾。
化合物经手工或者
Figure PCTCN2015082605-appb-000025
软件命名，市售化合物采用供应商目录名称。
用配有Shimadzu SIL-20A自动进样器和日本岛津DAD：SPD-M20A探测器的岛津LC20AB系统进行高效液相色谱分析，采用Xtimate C18(3m填料，规格为2.1x 300mm)色谱柱。0-60AB_6分钟的方法：应用线性梯度，以100％A(A为0.0675％TFA的水溶液)开始洗脱，并以60％B(B为0.0625％TFA的MeCN溶液)结束洗脱，整个过程为4.2分钟，然后以60％B洗脱1分钟。将色谱柱再平衡0.8分钟达到100:0，总运行时间为6分钟。10-80AB_6分钟的方法：应用线性梯度，以90％A(A为0.0675％TFA的水溶液)开始洗脱，并以80％B(B为0.0625％TFA的乙腈溶液)结束洗脱，整个过程为4.2分钟，然后以80％B洗脱1分钟。将色谱柱再平衡0.8分钟达到90:10，总运行时间为6分钟。柱温为50℃，流速为0.8mL/min。二极管阵列检测器扫描波长为200-400nm。
在Sanpont-group的硅胶GF254上进行薄层色谱分析(TLC)，常用紫外光灯照射检出斑点，在某些情况下也采用其他方法检视斑点，在这些情况下，用碘(10g硅胶中加入约1g碘并彻底混合而成)、香草醛(溶解大约1g香草醛于100mL 10％H2SO4中制得)、茚三酮(从Aldrich购得)或特殊显色剂(彻底混合(NH4)6Mo7O24·4H2O、5g(NH4)2Ce(IV)(NO3)6、450mL H2O和50mL浓H2SO4而制得)展开薄层板，检视化合物。采用Still,W.C.；Kahn,M.；and Mitra,M.Journal of Organic Chemistry,1978,43,2923-2925.中所公开技术的类似方法，在Silicycle的40-63μm(230-400目)硅胶上进行快速柱色谱。快速柱色谱或薄层色谱的常用溶剂是二氯甲烷/甲醇、乙酸乙酯/甲醇和己烷/乙酸乙酯的混合物。
在Gilson-281Prep LC 322系统上采用吉尔森UV/VIS-156探测器进行制备色谱分析,所采用的色谱 柱是Agella Venusil ASB Prep C18，5 m、、150x 21.2mm；Phenomenex Gemini C18、5m、150x 30mm；Boston Symmetrix C18，5 m、150x 30mm；或者Phenomenex Synergi C18、4m、150x 30mm。在流速约为25mL/min时，用低梯度的乙腈/水洗脱化合物，其中水中含有0.05％HCl、0.25％HCOOH或0.5％NH3·H2O，总运行时间为8-15分钟。
具体实施方式
为了更详细地说明本发明，给出下列实例，但本发明的范围并非限定于此。
流程A
Figure PCTCN2015082605-appb-000026
实施例1
(2-((5-氯-2-((2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000027
实施例1A
1-甲氧基-4-乙烯基苯
Figure PCTCN2015082605-appb-000028
1-溴-4-甲氧基苯(8.15克,43.6毫摩尔)，乙烯基三氟硼酸钾(7.08克,52.8毫摩尔)，Pd(dppf)Cl2(1.6克,2.20毫摩尔)和碳酸铯(28.7克,88.1毫摩尔)在1,4-二氧六环(120mL)和水(25mL)的混合物加热到110℃，并搅拌16小时。TLC显示反应完成，将反应混合物过滤，滤液浓缩至干，得到粗产物；粗产品通过通过柱色谱(PE)分离纯化，得到标题化合物(黄色油状，3.45克，产率59％)。
实施例1B
2,2-二氯-3-(4-甲氧基苯基)环丁酮
Figure PCTCN2015082605-appb-000029
实施例1A(3.45克，9.33毫摩尔)和铜-锌试剂(3.18克，64.3毫摩尔)在无水THF(25.0mL)的混合物加热到回流，然后依次滴加三氯氧磷(7.78克，50.7毫摩尔)和2,2,2-三氯乙酰氯(8.75克，48.1毫摩尔)的无水THF(25.0mL)溶液，滴加1小时后。将反应混合物回流下搅拌12小时。TLC(PE)显示反应完成，将混合物过滤，滤液减压浓缩，得到棕色油状化合物(6.0克，粗产物)，直接用于下一步反应。
实施例1C
3-(4-甲氧基苯基)环丁酮
Figure PCTCN2015082605-appb-000030
在搅拌下，向Zn粉(6.4克，98.4毫摩尔)的HOAc(25.0mL)的混合物中，滴加实施例1B(6.0克，24.6毫摩尔)的HOAc(25.0mL)溶液。反应混合物加热到70℃，并搅拌4小时。将反应混合物过滤，浓缩至干，得到粗产物，通过制备HPLC分离纯化，得到浅黄色油状物化合物(1.02克，产率24％)。1H NMR(400MHz,CDCl3):δ,7.23(dd,J＝6.8,1.6Hz,2H),6.91(dd,J＝9.6,2.8Hz,2H),3.82(s,3H),3.70-3.55(m,1H),3.54-3.40(m,2H),3.70-3.55(m,2H).
实施例1D
甲基2-(3-(4-甲氧基苯基)环丁基)乙
Figure PCTCN2015082605-appb-000031
在0℃下，向氢化钠(278毫克，6.96毫摩尔，60％)在无水THF(20.0mL)的悬浮液中，滴加甲基2-(二乙氧基磷酰基)乙酸乙酯(1.46克，6.96毫摩尔)，反应混合物在0℃搅拌0.5小时。然后实施例1C(1.02克，5.80毫摩尔)溶于无水THF(10mL)中，然后逐滴加入到上述反应混合物。将混合物升温至23℃，在23℃下搅拌16h。TLC(PE：乙酸乙酯＝10：1)显示反应完成。反应混合物冷却到0℃，慢慢加入水(20mL)淬灭反应，并将该混合物用EtOAc(20mL)萃取。有机层分别用水(10mL)和盐水(10mL)洗涤，然后用无水硫酸钠干燥，并浓缩至干，得到粗产物，将其通过柱色谱纯化(PE：乙酸乙酯＝10：1)，得到标题化合物(0.99克，73％)为无色油状物。1H NMR(400MHz,CDCl3):δ,7.21(d,J＝8.8Hz,2H),6.90(d,J＝8.8Hz,2H),5.80–5.68(m,1H),3.83(s,3H),3.73(s,3H),3.58-3.48(m,2H),3.33-3.16(m,2H),2.98-2.90(m,1H).
实施例1E
甲基2-(3-(4-甲氧基苯基)-1-(硝基甲基)环丁基)乙
Figure PCTCN2015082605-appb-000032
向实施例1D(0.99克，4.27毫摩尔)在无水THF(25.0mL)的混合物中加入硝基甲烷(521毫克，8.54毫摩尔)和TBAF(1.67克，6.41毫摩尔)。将反应混合物加热到70℃，并搅拌16小时。将混合物冷却至18℃，并浓缩至干，得到粗产物，将其通过柱色谱(PE：乙酸乙酯＝9:1)纯化，得到标题化合物(1.15g，产率92％)，为浅黄色油状物。1H NMR(400MHz,CDCl3):δ,7.18-7.08(m,2H),6.94-6.88(m,2H),4.89(s,1H),4.71(s,1H),3.82(s,3H),3.75(s,1.5H),3.71(s,1.5H),3.65-3.47(m,1H),2.90(s,1H),2.75(s,1H),2.65-2.57(m,1H),2.52-2.44(m,1H),2.32-2.26(m,1H),2.20-2.10(m,1H).
实施例1F
2-(4-甲氧基苯基)-6-氮杂螺[3.4]辛-7-酮
Figure PCTCN2015082605-appb-000033
在18℃氮气保护下，向实施例1E(1.15克，3.92毫摩尔)的甲醇(15.0mL)溶液中，加入Raney-Ni(0.20g)。反应混合物加热到50℃，在氢气中(压力：50Psi)搅拌16小时。LCMS显示反应完成。将反应混合物通过硅藻土过滤，滤液减压浓缩，得到标题化合物(0.77克，粗产品)为白色固体。LCMS(ESI)(10-80CD):m/z:232.2[M+1].
实施例1G
2-(4-甲氧基苯基)-6-甲基-6-氮杂螺[3.4]辛-7-酮
Figure PCTCN2015082605-appb-000034
在0℃下，向NaH(60毫克，1.5毫摩尔，60％)的无水DMF(5.0mL)混合物中，加入实施例1F(230毫克，1.0毫摩尔)；将反应混合物在0℃下搅拌1小时。然后加入MeI(213毫克，1.5毫摩尔，溶解于干燥的5mL无水THF中)逐滴加入到上述混合物中。将混合物升温至16℃并搅拌16小时。LCMS显示反应完全，向反应液中加入水(20mL)淬灭反应，并用乙酸乙酯(20mL)萃取。有机层分别用水(10mL)和盐水(10mL)洗涤，用无水硫酸钠干燥，浓缩至干，得到标题化合物(0.24克，粗品)为黄色油状物。LCMS(ESI)(5-95AB):m/z:246.3[M+1].
实施例1H
2-(4-甲氧基-3-硝基苯基)-6-甲基-6-氮杂螺[3.4]辛-7-酮
Figure PCTCN2015082605-appb-000035
将实施例1G(240毫克，1.0毫摩尔)的乙酸酐(5.0mL)溶液冷却到0℃，浓硝酸(1.0mL)慢慢加入到反应混合物中；将反应混合物升温至18℃，并搅拌2h。LCMS显示反应完全。将混合物过滤，滤液在减压条件下浓缩，得到标题化合物(202毫克，粗品)，为黄色油状物。LCMS(ESI)(5-95AB):m/z:291.2[M+1].
实施例1I
2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺
Figure PCTCN2015082605-appb-000036
在0℃下，向实施例1H(202毫克，0.70毫摩尔)的THF(10.0mL)溶液中，加入LAH(106毫克，2.80毫摩尔)；将反应混合物加热至70℃并搅拌12小时。LCMS显示反应完全。将反应混合物冷却至0℃，滴加乙酸乙酯(20.0mL)淬灭反应，随后加入水(2.0mL)；将混合物过滤，滤液在减压下浓缩，得到标题化合物(120mg，粗产物)为棕色油状物。LCMS(ESI)(10-80CD):m/z:247.2[M+1].
实施例1J
二甲基氧膦
Figure PCTCN2015082605-appb-000037
MeMgBr(1.30摩尔，434.46mL)的THF(800mL)溶液冷却至0℃，在N2保护下，二乙基膦酸酯(60.0克，434.46毫摩尔，溶解于40mL THF中)逐滴加入到反应混合物中，滴加2h并保持内部温度不高于0℃。加完后，将反应混合物升温至20℃并搅拌14小时。碳酸钾溶液(177克，溶解在250mL水中)加入到反应混合物中淬灭反应。有白色固体出现，将混合物过滤，滤饼用乙醇(100mL)洗涤，将滤液浓缩，并过滤浓缩过程中出现的固体。甲苯(200mL)加入到滤液中，并浓缩至干除掉多余水分，得到标题化合物(30.11克，385.78毫摩尔，88.79％产率)为无色粘稠的油状物。1HNMR(400MHz,CDCl3):δ,7.74-7.67(m,0.5H),6.57-6.52(m,0.5H),1.56(d,J＝3.6Hz,3H),1.53(d,J＝3.6Hz,3H).
实施例1K
(2-氨基苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000038
将2-碘苯胺溶液(12.50克，57.07毫摩尔)和实施例1J(5.35克，68.49毫摩尔)，K3PO4(14.54克，68.49毫摩尔)，Xantphos(660.44毫克，1.14毫摩尔)和醋酸钯(256.26毫克，1.14毫摩尔)置于DMF(80mL)中，反应混合物在氮气保护下，加热至100℃并搅拌16小时。LCMS(DCM：甲醇＝10：1)显示反应完成。将混合物过滤并浓缩，将所得残余物用HCl水溶液(1N，80mL)稀释，调节pH值至大约2，将所得混合物过滤。滤液用DCM(100mL×2)萃取，将水层分离出来，并且用碳酸氢钠水溶液将pH值调节至约9，然后用DCM(200mL×2)萃取。有机层用无水硫酸钠干燥并浓缩至干。将粗产物通过重结晶(PE：乙酸乙酯＝5:1)纯化，得到标题化合物(6.00克，35.47毫摩尔，62.15％产率) 为白色固体。1H NMR(400MHz,CDCl3):δ,7.20(t,J＝7.6Hz,1H),7.04(dd,J＝13.6,7.6Hz,1H),6.69-6.58(m,2H),5.35(br s,2H),1.75(s,3H),1.71(s,3H).LCMS(ESI)(10-80CD):m/z:170.1[M+1].
实施例1L
(2-((2,5-二氯嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000039
在16℃下，向实施例1K(2.50克，14.8毫摩尔)和2，4，5-三氯嘧啶(2.85克，15.5毫摩尔)的DMF(20mL)混合物中，加入DIPEA(3.82克，29.6毫摩尔)。然后将反应混合物加热至70℃并搅拌16小时。TLC显示反应完成。将反应混合物用水(50mL)稀释，并用EtOAc(40mL×3)萃取。将合并的有机相用饱和盐水(20mL×2)洗涤，无水硫酸钠干燥，过滤并真空浓缩。粗产物在乙醇中重结晶，得到标题化合物(3.20克，10.1毫摩尔，68.4％产率)为白色固体。1H NMR(400MHz,CD3OD):δ,8.50(dd,J＝8.0,4.0Hz,1H),8.35-8.28(m,1H),7.69-7.59(m,2H),7.36-7.28(m,1H),1.91(s,3H),1.88(s,3H).LCMS(ESI)(5-95AB):m/z:315.9[M+1].
实施例1M
(2-((5-氯-2-((2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000040
向实施例1I(120毫克，0.49毫摩尔)，实施例1L(154毫克，0.49毫摩尔)在叔丁醇(5.0mL)的混合物中，加入MeSO3H(141毫克，1.47毫摩尔)；将反应混合物在氮气保护下加热至90℃反应12小时。LCMS显示反应完全，减压浓缩后，得到粗产物，将其通过制备HPLC分离纯化，得到标题化合物(52.2毫克，收率20％)为棕色油状物。1H NMR(400MHz,CD3OD):δ,8.45(s,1H),8.31-8.27(m,1H),8.10(s,1H),7.82(s,1H),7.70-7.64(m,1H),7.57-7.51(m,1H),7.34-7.30(m,1H),6.95-6.89(m,2H),3.86(s,3H),3.53(s,1H),3.44(s,1H),3.35-3.32(m,1.5H),3.28-3.22(m,1.5H),2.96(s,1.5H),2.88(s,1.5H),2.44-2.39(m,1H),2.36-2.27(m,2H),2.21-2.14(m,2H),2.12-2.08(m,1H),1.87(s,3H),1.84(s,3H).LCMS(ESI)(5-95AB):m/z:526.2[M+1].
实施例2
2-(3-((5-氯-4-((2-(二甲基膦酰基)苯基)氨基)嘧啶-2-基)氨基)-4-甲氧基苯基)-6-甲基-6-氮杂螺[3.4]辛烷7-酮
Figure PCTCN2015082605-appb-000041
实施例2A
2-(3-氨基-4-甲氧基苯基)-6-甲基-6-氮杂螺[3.4]辛烷-7-酮
Figure PCTCN2015082605-appb-000042
在氩气保护下，向实施例1H(701毫克，2点42毫摩尔)的MeOH(5.0mL)溶液中，加入Pd/C(150毫克)；反应混合物在H2(压力：15psi)下于22℃反应16小时。LCMS显示反应的完成，反应混合物通过硅藻土过滤，滤液减压浓缩，得到标题化合物(596毫克，粗品)为白色固体。LCMS(ESI)(10-80CD):m/z:261.2[M+1].
实施例2B
2-(3-((5-氯-4-((2-(二甲基膦酰基)苯基)氨基)嘧啶-2-基)氨基)-4-甲氧基苯基)-6-甲基-6-氮杂螺[3.4]辛烷7-酮
Figure PCTCN2015082605-appb-000043
本实施例制备方法与实施例1M中描述的方法一致，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛烷-2-基)苯胺用2-(3-氨基-4-甲氧基苯基)-6-甲基-6-氮杂螺[3.4]辛烷-7-酮代替。得到的标题化合物为棕色油状物(32％产率)。LCMS(ESI)(5-95AB):m/z:540.1[M+1].
流程B
Figure PCTCN2015082605-appb-000044
实施例3
(2-((5-氯-2-((2-甲氧基-4-(2,8-二氮杂螺[4.5]癸烷-8-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000045
实施例3A
叔丁基4-(2-乙氧基-2-氧代亚乙基)哌啶-1-甲酸叔丁酯
Figure PCTCN2015082605-appb-000046
在0℃下，向乙基2-(二乙氧基磷酰基)乙酸乙酯(6.18克，27.6毫摩尔)的无水THF(100mL)溶液中，慢慢分批加入NaH(1.2克，30.1毫摩尔，60％)，该混合物在0℃下搅拌1小时，然后叔丁基-4-氧代哌啶-1-甲酸叔丁酯(5克，25.1毫摩尔)缓慢加入到反应混合物中。反应混合物在0℃下搅拌12小时。TLC(石油醚：乙酸乙酯＝3：1)显示反应完成。向反应液中加入水(50mL)淬灭反应，用EtOAc(100mL)萃取，无水硫酸钠干燥有机层，并浓缩至干，得到标题化合物(4.95克，产率73％) 为白色固体。1H NMR(400MHz,CDCl3):δ,5.73(s,1H),4.23-4.14(m,2H),3.53-3.47(m,4H),2.96-2.93(m,2H),2.31-2.28(m,2H),1.48(s,9H),1.28-1.32(m,3H).
实施例3B
叔丁基4-(2-乙氧基-2-氧代乙基)-4-(硝基甲基)哌啶-1-甲酸叔丁酯
Figure PCTCN2015082605-appb-000047
在30℃下，向实施例3A(5克，18.6毫摩尔)的MeCN(100mL)反应混合物中，加入DBU(5.62克，37.0毫摩尔)和硝基甲烷(2.3克，37毫摩尔)，反应混合物加热至80℃并搅拌12小时，TLC(PE：乙酸乙酯＝3：1)显示反应完成。将反应混合物浓缩以除去溶剂；得到的残留物用EtOAc(50mL)稀释，并依次用水(30mL)，饱和NaHCO 3(20mL)和盐水(20mL)洗涤；无水硫酸钠干燥有机层，并浓缩得到棕色油状物，用柱色谱(PE：乙酸乙酯＝10:1，8:1)分离纯化，得到标题化合物(3.2克，产率52.4％)为无色油状物。1H NMR(400MHz,CDCl3):δ,4.76(s,2H),4.24-4.16(q,J＝7.2Hz,2H),3.57-3.52(m,2H),3.44-3.39(m,2H),2.64(s,2H),1.68-1.63(m,4H),1.48(s,9H),1.30(t,J＝7.2Hz,3H).LCMS(ESI)(5-95AB):m/z:353.1[M+Na+].
实施例3C
叔-丁基-3-氧代-2,8-二氮杂螺[4.5]癸烷-8-羧酸乙酯
Figure PCTCN2015082605-appb-000048
实施例3B(1.5克，4.54毫摩尔)的甲醇(50mL)溶液搅拌10分钟，然后在氩气下保护下，加入Raney Ni(150毫克，10％)。反应混合物在氢气(压力：50psi)下加热到50℃搅拌12小时。TLC(PE：乙酸乙酯＝10：1)显示反应完成。反应混合物过滤并浓缩，得到标题化合物(920毫克，收率80％)为白色固体。LCMS(ESI)(5-95AB):m/z:509.4[2M+1].
实施例3D
2,8-二氮杂螺[4.5]癸烷-3-酮
Figure PCTCN2015082605-appb-000049
实施例3C(920毫克，3.62毫摩尔)的DCM/TFA(5mL/5mL)混合物，在30℃下搅拌12小时，TLC(PE：乙酸乙酯＝1:1)显示完成。反应混合物在减压下浓缩得到粗产物，该粗产物溶解在水(30mL)中，用氢氧化钠溶液(1N)将pH调至10；该混合物用二氯甲烷萃取(15mL×4)。合并的有机层用无水硫酸钠干燥并浓缩，得到标题化合物(550mg，产率98％)为黄色油状物。
实施例3E
8-(3-甲氧基-4-硝基苯基)-2,8-二氮杂螺[4.5]癸烷-3-酮
Figure PCTCN2015082605-appb-000050
向实施例3D(550毫克，3.57毫摩尔)的DMF(15mL)溶液中，加入碳酸钾(1.5克，10.7毫摩尔)和4-氟-2-甲氧基-1-硝基苯(855毫克，5.0毫摩尔；然后，反应混合物加热至90℃并搅拌12小时。TLC(DCM：甲醇＝20：1)显示反应完成。反应混合物用EtOAc(50mL)稀释，依次用水(20mL)，饱和NaHCO3(20mL)，盐水(20mL)洗涤，有机相用无水Na2SO4干燥，然后过滤并浓缩得到棕色油状物，粗品通过柱色谱(DCM：甲醇＝20:1)分离纯化，得到标题化合物(1.02克，产率95％)为黄色固体。1H NMR(400MHz,DMSO-d6):δ,7.89(d,J＝9.6Hz,1H),7.59(s,1H),6.60(dd,J＝2.4Hz,9.6Hz,1H),6.51(d,J＝2.4Hz,1H),3.90(s,3H),3.59-3.56(m,2H),3.44-3.42(m,2H),3.17(d,J＝5.2Hz,,2H),3.10(s,2H),1.64-1.61(t,J＝5.6Hz,4H).LCMS(ESI)(5-95AB):m/z:306.3[M+1].
实施例3F
8-(4-氨基-3-甲氧基苯基)-2,8-二氮杂螺[4.5]癸烷-3-酮
在30℃下，向实施例3E(500毫克，1.64毫摩尔)的EtOH(10mL)和水(10mL)的混合物中，加入铁粉(367毫克，6.56毫摩尔)与氯化铵(260毫克，4.92毫摩尔)。该反应混合物加热至80℃搅拌5小时，TLC(DCM：甲醇＝20：1)显示反应完成。反应混合物过滤，滤液在真空下浓缩，得到标题化合物(450毫克，产率99％)为黑色油状物。
实施例3G
2-甲氧基-4-(2,8-二氮杂螺[4.5]癸烷-8-基)苯胺
Figure PCTCN2015082605-appb-000052
实施例3F(400毫克，1.45毫摩尔)的无水THF(10mL)溶液在0℃下搅拌0.5小时，LiAlH4(58毫克1.53毫摩尔)分批次加入到上述混合物中。加完后，将混合物加热至80℃反应12小时。TLC(DCM：甲醇＝6:1)显示反应完成。向反应混合物中加入水(10mL)淬灭，并用DCM萃取(10mL×3)，有机相分离，干燥并浓缩，得到粗产物为黑色油状物，粗产品通过制备HPLC分离纯化，得到标题化合物(80mg，产率20％)为白色固体。
实施例3H
(2-((5-氯-2-((2-甲氧基-4-(2,8-二氮杂螺[4.5]癸烷-8-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000053
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为2-甲氧基-4-(2,8-二氮杂螺[4.5]癸烷-8-基)苯胺。得到标题化合物为白色固体(产率20％)。LCMS(ESI)(5-95AB):m/z:541.1[M+1].
实施例4
(2-((5-氯-2-((2-甲氧基-4-(2,7-二氮杂螺[3.5]壬烷-7-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000054
实施例4A
叔丁酯7-(4-甲氧基-3-硝基苯基)-2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯
Figure PCTCN2015082605-appb-000055
2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(100毫克，0.44毫摩尔)，4-氟-2-甲氧基-1-硝基苯(113毫克，0.66毫摩尔)和碳酸钾(152毫克，1.1毫摩尔)的DMSO(3mL)混合物，加热到90℃并搅拌12小时。LCMS显示反应完成，向反应混合物加入水(20mL)，并用DCM(25mL×2)萃取。将有机层用盐水洗涤，用无水硫酸钠干燥，过滤并浓缩，得到残余物，该粗品用制备TLC分离纯化(PE：乙酸乙酯＝1:1)得到标题化合物(150毫克，产率90％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:378.1[M+1].
实施例4B
叔丁酯7-(3-氨基-4-甲氧基苯基)-2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯
Figure PCTCN2015082605-appb-000056
本实施例根据实施例3F的方法制备，将8-(3-甲氧基-4-硝基苯基)-2,8-二氮杂螺[4.5]癸烷-3-酮替换为叔丁酯7-(4-甲氧基-3-硝基苯基)-2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯。得到标题化合 物，为绿色油状物(粗品)，直接用于下一步骤。LCMS(ESI)(5-95AB):m/z:348.2[M+1].
实施例4C
(2-((5-氯-2-((2-甲氧基-4-(2,7-二氮杂螺[3.5]壬烷-7-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000057
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为叔丁酯7-(3-氨基-4-甲氧基苯基)-2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯。得到标题化合物为无色油状化合物(收率16％)。LCMS(ESI)(5-95AB):m/z:527.1[M+1].
实施例5
(2-((5-氯-2-((2-甲氧基-4-(2-甲基-2,7-二氮杂螺[3.5]壬烷-7-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000058
实施例5A
1-叔丁基-4-甲基-4-(氯甲基)哌啶-1,4-二羧酸酯
Figure PCTCN2015082605-appb-000059
在N2保护，-78℃下，向1-叔丁基-4-甲基哌啶-1,4-二羧酸酯(4.60克，18.91毫摩尔，1.00当量)的四氢呋喃(100mL)溶液中，逐滴加入LDA(2M，18.91mL)溶液。滴加完成后，混合物在-78℃和氮气保护下搅拌2小时。然后在-78℃下，通过一个注射器将氯碘甲烷(10克，56.72毫摩尔)逐滴加入到反应混合溶液中。然后，将所得溶液慢慢升温至20℃并搅拌12小时。反应体系用氯化铵水溶液淬灭，并向反应体系中加入乙酸乙酯和水。将有机层分离并浓缩，粗品经柱色谱(石油醚：乙酸乙酯＝100：1到25：1)分离纯化，得到标题化合物(2.39克，8.19毫摩尔，43.32％产率)为黄色油状物。1H NMR(400MHz,CDCl3):3.86(br.s.,2H)3.76(s,3H)3.59(br.s.,2H)3.00(br.s.,2H),2.16(d,J＝13.2Hz,2H)1.45(m,9H).
实施例5B
叔丁基4-(氯甲基)-4-(羟甲基)哌啶-1-甲酸叔丁酯
Figure PCTCN2015082605-appb-000060
实施例5A(14.4克，49.35毫摩尔)的无水四氢呋喃溶液(150mL)冷却至0℃，LAH(2.25克，59.22毫摩尔)分批次缓慢加入到溶液中，并将该混合液在0℃下搅拌25分钟。TLC显示反应完全，混合物在0-10℃下，用水(2.25mL)淬灭，然后加入氢氧化钠溶液(1N，2.25mL)，混合物过滤；滤饼用乙酸乙酯洗涤(30mL×2)，然后滤液用水(150mL)和盐水(150mL)洗涤，无水硫酸钠干燥，浓缩，得到标题化合物(9.79克，37.12毫摩尔，75.21％产率)为淡黄色油状物。1H NMR(400MHz,CDCl3):3.69-3.56(m,4H),3.47-3.37(m,4H),1.60-1.51(m,4H),1.50-1.43(m,9H).
实施例5C
叔丁基4-(氯甲基)-4-甲酰基哌啶-1-甲酸叔丁酯
Figure PCTCN2015082605-appb-000061
在-65℃下，向草酰氯(5.34克，42.07毫摩尔)的二氯甲烷溶液(90mL)中加入二甲基亚砜(6.57克，84.15毫摩尔)的二氯甲烷(20mL)溶液。然后，实施例5B(9.79克，37.12毫摩尔)的二氯甲烷溶液(20mL)逐滴加入到上述混合物中，并保持内部温度低于-60℃。反应混合物在-65℃至-60℃下搅拌15分钟。然后在-60℃下，向反应混合物中加入三乙胺(18.77克，185.49毫摩尔)。待加完后，将反应混合物升温至室温(20℃)，用饱和碳酸氢钠水溶液(50mL)淬灭。将有机层分离，并用盐水(30mL)洗涤，用无水硫酸钠干燥，并通过柱色谱(二氯甲烷：甲醇＝50：1至10：1)分离纯化，得到标题化合物(10.50克，粗品)为浅黄色油。1H NMR(400MHz,CDCl3):9.59(s,1H),3.92-3.67(m,2H),3.62(s,2H),3.10(t,J＝10.4Hz,2H),2.08(dt,J＝13.6,4.0Hz,2H),1.60–1.52(m,2H),1.47(s,9H).
实施例5D
叔丁基-2-甲基-2,7-二氮杂螺[3.5]壬烷-7-羧酸乙酯
Figure PCTCN2015082605-appb-000062
在15℃下，向实施例5C(10.50克，40.12毫摩尔)和甲胺盐酸盐(10.51克，155.65毫摩尔)的甲醇(100mL)混合液中加入氰基硼氢化钠(14.50克，230.7毫摩尔)，然后，混合物在100℃下搅拌16小时。TLC显示反应完全。将混合物浓缩至干，得到的残余物用乙酸乙酯(100mL)稀释，用水(100mL)和盐水(100mL)洗涤，用无水硫酸钠干燥并浓缩。粗产物经柱色谱(二氯甲烷：甲醇＝100：1至10:1)分离纯化，得到标题化合物(4.20克，17.48毫摩尔，43.56％产率)为黄色油状物。1H NMR(400MHz,CDCl3):3.97(br.s.,4H),3.42-3.36(m,4H),2.99(s,3H),1.94-1.84(m,4H),1.45(s,9H).
实施例5E
2-甲基-2,7-二氮杂螺[3.5]壬烷
Figure PCTCN2015082605-appb-000063
在15℃下，实施例5D(4.20克，17.48毫摩尔)的盐酸/甲醇(30mL)溶液搅拌0.5小时。TLC显示反应完全。将混合物浓缩，得到标题化合物(3.62克，盐酸盐)为白色固体。1H NMR(400MHz,CD3OD):4.24(d,J＝15.6Hz,2H),4.00(d,J＝15.2Hz,2H),3.29-3.15(m,4H),2.99(s,3H),2.32-2.07(m,4H).
实施例5F
7-(3-甲氧基-4-硝基苯基)-2-甲基-2,7-二氮杂螺[3.5]壬烷
Figure PCTCN2015082605-appb-000064
本实施例根据实施例4A的方法制备，将叔丁酯2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯替换为2-甲基-2,7-二氮杂螺[3.5]壬烷。得到标题化合物为黄色固体，产率92％。LCMS(ESI)(10-80CD):m/z:292.2[M+1].
实施例5G
2-甲氧基-4-(2-甲基-2,7-二氮杂螺[3.5]壬-7-基)苯胺
Figure PCTCN2015082605-appb-000065
在15℃下，向实施例5F(1.20克，4.12毫摩尔)和氯化铵(1.40克，26.17毫摩尔)的甲醇(20mL)和二氯甲烷(2mL)的混合液中中加入锌粉(2.00克，30.59毫摩尔)，然后，反应混合液在35℃下搅拌10分钟。TLC显示反应完成。将混合物过滤，将滤液浓缩。将浓缩残留物溶解在饱和的碳酸钾溶液(20mL)中，并用二氯甲烷和甲醇的混合物萃取(二氯甲烷：甲醇＝20:1，20mL*2)。将合并的有机层用无水硫酸钠干燥并浓缩，得到标题化合物(920毫克，3.52毫摩尔，85.44％产率)为深绿色油状物。1H NMR(400MHz,CDCl3):6.64(d,J＝8.4Hz,1H),6.53(d,J＝2.4Hz,1H),6.42(dd,J＝8.4,2.4Hz,1H),3.85(s,3H),3.07(s,4H),2.97-2.92(m,4H),2.37(s,3H),1.95-1.85(m,4H).
实施例5H
(2-((5-氯-2-((2-甲氧基-4-(2-甲基-2,7-二氮杂螺[3.5]壬烷-7-基)苯基)氨基)嘧啶-4- 基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000066
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为2-甲氧基-4-(2-甲基-2,7-二氮杂螺[3.5]壬-7-基)苯胺。得到标题化合物为黄色固体，产率23％。1H NMR(400MHz,CD3OD):8.22(s,2H),7.87-7.61(m,3H),7.57-7.45(m,1H),7.38(br.s.,1H),7.12(d,J＝8.0Hz,1H),4.34(d,J＝10.4Hz,2H),4.06(d,J＝11.2Hz,2H),3.99(s,3H),3.74-3.49(m,4H),3.03(s,3H),2.51–2.30(m,4H),1.89(d,J＝13.6Hz,6H)；LCMS(ESI)(0-60AB):m/z:541.2[M+1].
实施例6
(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000067
实施例6A
叔丁基6-(4-甲氧基-3-硝基苯基)-2,6-二氮杂螺[3.4]辛烷-2-甲酸叔丁酯
Figure PCTCN2015082605-appb-000068
本实施例根据实施例4A的方法制备，将叔丁酯2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯替换为叔丁酯2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯。得到标题化合物为无色油状化合物，收率92％。LCMS(ESI)(5-95AB):m/z:364.1[M+1].
实施例6B
叔丁基6-(3-氨基-4-甲氧基苯基)-2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯
Figure PCTCN2015082605-appb-000069
本实施例根据实施例3F的方法制备，将8-(3-甲氧基-4-硝基苯基)-2,8-二氮杂螺[4.5]癸烷-3- 酮替换为叔丁基6-(4-甲氧基-3-硝基苯基)-2,6-二氮杂螺[3.4]辛烷-2-甲酸叔丁酯。得到标题化合物为绿色油状物(粗品)。LCMS(ESI)(5-95AB):m/z:334.2[M+1].
实施例6C
(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000070
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为叔丁基6-(3-氨基-4-甲氧基苯基)-2,6-二氮杂螺[3.4]辛烷-2-羧酸叔丁酯。得到标题化合物为无色油状物，产率19％。LCMS(ESI)(5-95AB):m/z:513.0[M+1].
实施例7
(2-((5-氯-2-((2-甲氧基-4-(2-甲基-2,6-二氮杂螺[3.4]辛烷-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000071
向实施例6C(50毫克，0.039毫摩尔)的四氢呋喃溶液(5mL)中加入甲醛水溶液(9.5毫克，37％，0.117毫摩尔)；反应混合物在16℃下搅拌30分钟，然后向反应混合物中加入醋酸硼氢化钠(25毫克，0.117毫摩尔)，并在16℃下搅拌12小时。LCMS显示反应完成。将反应混合物过滤，通过制备HPLC纯化，得到标题化合物(9.6毫克，产率46％)为白色固体。LCMS(ESI)(5-95AB):m/z:527.2[M+1].
实施例8
(2-((5-氯-2-((2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000072
实施例8A
9-苄基-2,4-二氧代-3,9-二氮杂螺[5.5]十一烷-1,5-二腈
Figure PCTCN2015082605-appb-000073
在5-8℃下，醋酸铵(2.04克，26.42毫摩尔，0.10当量)加入到氰基乙酸酯(90克，796毫摩尔，3.00当量)的甲醇溶液(100mL)中；然后1–苄基哌啶4-酮(50克，0.264摩尔)加入到上述反应混合物中；然后，在10℃以下，氨水(46.3克，370毫摩尔，1.40当量)加入到反应混合物中，将该混合物在0-5℃下搅拌1小时。然后反应混合物升温至20℃(室温)，并搅拌20小时。LCMS显示生成产物。水(100mL)加入到混合物中，并加热至55℃。加入浓盐酸(12M)将pH调至4，并保持温度不超过70℃。然后将反应液冷却至10℃，搅拌30分钟后过滤。滤饼用水洗涤，放置空气中干燥，得到标题化合物(66克，77％产率)为白色固体。LCMS(ESI)(0-30AB):m/z:323.0[M+1].
实施例8B
1'-苄基-3,7-二氮杂螺[二环[3.3.1]壬烷-9，4'-哌啶]-2,4,6,8-四酮
Figure PCTCN2015082605-appb-000074
实施例8A(1.00克，3.10毫摩尔，1.00当量)的硫酸(88％，4mL)混合液在60℃下搅拌4小时。然后向反应液中加入水(1.4mL)，并加热到100℃搅拌1小时。再向反应混合物中加入水(5mL)，并冷却至10℃，并在10℃下搅拌30分钟，然后过滤。滤饼用冷水(5mL)洗涤并干燥，得到标题化合物(1.11克，粗品)为白色固体。1H NMR(400MHz,CDCl3):11.87(s,2H),9.56(br.s.,1H),7.47(s,5H),4.34(d,J＝4.4Hz,2H),3.75(br.s.,1H),3.22(br.s.,4H),1.88(br.s.,4H).
实施例8C
9-苄基-3,9-二氮杂螺[5.5]十一烷-2,4-二酮
Figure PCTCN2015082605-appb-000075
在15℃下，向实施例8B(1.10克，3.22毫摩尔，1.00当量)的烧瓶中加入NaOH水溶液(5N，5mL)，然后，将混合物升温至70℃并搅拌4小时。将混合液冷却至45℃，然后将浓盐酸(12N，～1.5mL)慢慢地加入，直到溶液的pH值为7左右。再将混合物加热至70-75℃，然后将浓HCl(12N，～1mL)逐滴加入，以控制二氧化碳释放的速度，直到pH值调至3-4左右。将混合物加热到70-75℃并进一步反应1小时。将所得悬浮液冷却至10℃并搅拌0.5小时。将固体过滤并用水洗涤(25mL)中。将固体干燥，得到标题化合物(380毫克，1.40毫摩尔，2步产率45％)为白色固体。1H NMR(400MHz,CDCl3): 10.89(s,1H),10.60(br.s,1H),7.57(br.s.,2H),7.44(br.s.,3H),4.28(br.s.,2H),3.11(br.s.,4H),2.76(br.s.,2H),2.42(br.s.,2H),2.00-1.48(m,4H).
实施例8D
3-苄基-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000076
在0-10℃下，向实施例8C(10.6克，38.92毫摩尔，1.00当量)的四氢呋喃溶液(120mL)中加入氢化铝锂(5.17克，136.22毫摩尔，3.50当量)，然后将混合液在65℃下搅拌下3小时。TLC显示反应完成。将混合液冷却至10℃，并加入水(5.2mL)淬灭反应，接着加入氢氧化钠水溶液(1N，5.2mL)。将混合物过滤，滤液浓缩，得到标题化合物(7.40克，30.28毫摩尔，77.81％产率)为淡黄色油状物。1H NMR(400MHz,CDCl3):7.34-7.27(m,5H),3.52(s,2H),2.88-2.73(m,4H),2.46-2.35(m,4H),1.58-1.51(m,4H),1.48-1.39(m,4H).
实施例8E
叔丁酯3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000077
向实施例8D(500毫克，2.05毫摩尔)和Boc2O(450毫克，2.06毫摩尔)的甲醇混合液中加入三乙胺(311毫克，3.08毫摩尔)，然后将混合物在20-30℃下搅拌16小时。将反应混合物浓缩，残余物用乙酸乙酯(20mL)稀释，并用水(15mL×2)和盐水(20mL)洗涤，将有机层用无水硫酸钠干燥，浓缩。将粗中间体溶于乙醇(15mL)和醋酸(2mL)中，然后加入氢氧化钯/碳(0.1克)，并将该混合物在氢气(50Psi)下反应20小时。将混合物过滤，滤液浓缩，得到标题化合物的乙酸盐(320毫克，1.26毫摩尔，61.37％产率)。
实施例8F
叔丁酯9-(3-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000078
本实施例根据实施例4A的方法制备，将叔丁基2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯替换为叔丁基3,9-二氮杂螺[5.5]十一烷-3-甲酸叔丁酯。得到标题化合物为黄色油状物，产率62％。1H NMR(400MHz,CD3OD):δ,7.95(d,J＝9.2Hz,1H),6.55(dd,J＝9.6,2.4Hz,1H),6.48(d,J＝2.4Hz,1H),3.95(s,3H),3.56-3.41(m,8H),1.72-1.65(m,4H),1.58-1.50(m,4H),1.48(s,9H).
实施例8G
叔丁酯9-(4-氨基-3-甲氧基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000079
本实施例根据实施例3F的方法制备，将8-(3-甲氧基-4-硝基苯基-2,8-二氮杂螺[4.5]癸烷-3-酮替换为叔丁酯9-(3-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸乙酯。得到标题化合物为棕色油，收率36％。LCMS(ESI)(5-95AB):m/z:376.2[M+1].
实施例8H
(2-((5-氯-2-((2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000080
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为叔丁酯9-(4-氨基-3-甲氧基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯。得到标题化合物为黄色固体，产率21％。LCMS(ESI)(5-95AB):m/z:555.2[M+1].
实施例9
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000081
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((5-氯-2-((2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦。得到标题化合物为黄色固体，产率57％。1H NMR(400MHz,CD3OD):δ,8.28(s,1H),8.12(br.s.,1H),7.81-7.68(m,3H),7.65(d,J＝2.0Hz,1H),7.56-7.49(m,1H),7.37(d,J＝8.8Hz,1H),4.02(s,3H),3.74(br.s.,4H),3.47(d,J＝12.8Hz,2H),3.24(t,J＝12.8Hz,2H),2.93(s,3H),2.42-1.97(m,5H),1.93-1.75(m,9H).LCMS(ESI)(0-60AB):m/z:569.2[M+1].
实施例10
(2-((5-氯-2-((4-(9-异丙基-3,9-二氮杂螺[5.5]十一烷-3-基)-2-甲氧基苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000082
实施例8C(100毫克，0.18毫摩尔)，醋酸硼氢化钠(114毫克，0.54毫摩尔)，乙酸(21.6毫克，0.36毫摩尔)和丙酮(20.9毫克，0.36毫摩尔)的四氢呋喃(5.0mL)混合物，在18℃下搅拌16小时。LCMS显示反应完全。减压浓缩除去溶剂，得到粗产物，将其通过制备HPLC纯化，得到标题化合物(50mg，产率47％)为棕色固体。LCMS(ESI)(10-80AB):m/z:597.3[M+1].
实施例11
(2-((5-氯-2-((2-(二氟甲氧基)-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000083
实施例11A
5-氟-2-硝基苯酚
Figure PCTCN2015082605-appb-000084
在0℃下，向搅拌的4-氟-2-甲氧基硝基苯(3克，17.53毫摩尔)的二氯甲烷(30mL)的溶液中滴加三溴化硼。反应液在0℃下搅拌1.5小时。TLC(石油醚：乙酸乙酯＝10：1)显示4-氟-2-甲氧基硝基苯消失。将溶液缓慢加入到冰水(100mL)中，并用二氯甲烷(50mL×3)萃取。将有机相干燥并浓缩，得到标题化合物(2.5g，收率90.8％)为黄色油状物。1H NMR(400MHz,CDCl3):δ,10.80(s,1H),8.16(dd,J＝9.6,5.6Hz,1H),6.84(dd,J＝9.6,2.4Hz,1H),6.77-6.67(m,1H).
实施例11B
2-(二氟甲氧基)-4-氟-1-硝基苯
Figure PCTCN2015082605-appb-000085
在不断搅拌下，向实施例11A(2.0克，12.73毫摩尔)的N,N-二甲基甲酰胺(20mL)溶液中加入ClCF2COONa(6.9克，44.56毫摩尔)和碳酸钠(1.62克，15.28毫摩尔)。将反应混合液加热至90℃并搅拌16小时。TLC(石油醚：乙酸乙酯＝10：1)显示5-氟-2-硝基苯酚消失。将反应液用乙酸乙酯稀释(100mL)，并用水(20mL×2)洗涤。有机层干燥并浓缩，得到粗产物，将其通过硅胶柱纯化(石油醚：乙酸乙酯＝10：1)，得到标题化合物(1.4g，收率53.1％)为黄色油状物。1H NMR(400MHz,CDCl3):δ,8.07-7.98(m,1H),7.17-7.05(m,2H),6.65(t,J＝72.0Hz,1H).
实施例11C
叔丁酯9-(3-(二氟甲氧基)-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000086
本实施例根据实施例4A的方法制备，将叔丁酯2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯替换为叔丁酯3,9-二氮杂螺[5.5]十一烷-3-甲酸叔丁酯，以及将4-氟-2-甲氧基-1-硝基苯替换为2-(二氟甲氧基)-4-氟-1-硝基苯。得到标题化合物为黄色油状物，产率77％。
实施例11D
叔丁酯9-(4-氨基-3-(二氟甲氧基)苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000087
本实施例根据实施例3F的方法制备，将8-(3-甲氧基-4-硝基苯基-2,8-二氮杂螺[4.5]癸烷-3-酮替换为叔丁酯9-(3-(二氟甲氧基)基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸乙酯。得到标题化合物为绿色固体，产率95％。LCMS(ESI)(5-95AB):m/z:412.2[M+1].
实施例11E
(2-((5-氯-2-((2-(二氟甲氧基)-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000088
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺 替换为叔丁酯9-(4-氨基-3-(二氟甲氧基)苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯。标题化合物为白色固体，产率22％。LCMS(ESI)(0-60AB):m/z:591.2[M+1].
实施例11F
(2-((5-氯-2-((2-(二氟甲氧基)-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000089
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((5-氯-2-((2-(二氟甲氧基)-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦。得到标题化合物为无色油，收率22％。1H NMR(400MHz,CD3OD):δ,8.31(s,1H),8.12(m,1H),7.86-7.84(m,1H),7.79-7.65(m,3H),7.62-7.55(m,1H),7.54-7.48(m,1H),7.08(t,J＝72.0Hz,1H),3.75-3.62(m,4H),3.52-3.45(m,2H),3.29-3.21(m,2H),2.94(s,3H),2.34-2.13(m,4H),2.11-1.98(m,2H),1.89(d,J＝13.6Hz,6H),1.88-1.79(m,2H).LCMS(ESI)(5-95AB):m/z:605.2[M+1].
实施例12
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基硫醚
Figure PCTCN2015082605-appb-000090
实施例12A
(2-氨基苯基)二甲基硫醚
Figure PCTCN2015082605-appb-000091
(2-氨基苯基)二甲基氧膦(1.00克，5.91毫摩尔)和劳森试剂(4.78克，11.82毫摩尔)置于甲苯(60mL)中，反应混合物加热到110℃并搅拌4小时。TLC(石油醚：乙酸乙酯＝3：1)显示反应完成。将混合物过滤并浓缩，粗产物通过制备HPLC(碱性)纯化，得到标题化合物(600毫克，3.24 毫摩尔，产率54.81％)为无色油状物。1H NMR(400MHz,CDCl3):δ,7.31-7.27(m,1H),7.22-7.15(m,1H),6.81-6.75(m,1H),6.68-6.60(m,1H),2.08(s,3H),2.05(s,3H)
实施例12B
(2-((2,5-二氯嘧啶-4-基)氨基)苯基)二甲基硫醚
Figure PCTCN2015082605-appb-000092
实施例12A(50.0毫克，269.94微摩尔)，2,4,5-三氯嘧啶(148.54毫克，809.82微摩尔)和碳酸钾(111.93毫克，809.82微摩尔)置于N,N-二甲基甲酰胺(3mL)中，反应混合物在60℃下搅拌12小时。TLC(石油醚：乙酸乙酯＝3:1)显示产品生成。向该反应混合物中加入水(10mL)，并用乙酸乙酯(10mL×3)萃取。将合并的有机层干燥并浓缩，得到黄色油状物。该油状物通过制备TLC(石油醚：乙酸乙酯＝3:1)分离纯化，得到标题化合物(25.00毫克，75.26微摩尔，27.88％产率)为黄色固体。1H NMR(400MHz,CDCl3):δ,10.12(br.s.,1H),8.28(s,1H),8.10(dd,J＝8.0,4.8Hz,1H),7.61(dd,J＝8.0,8.0Hz,1H),7.46(dd,J＝13.8,7.6Hz,1H),7.33-7.28(m,1H),2.07(s,3H),2.04(s,3H).
实施例12C
3-(3-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000093
实施例8A(0.7克，1.73毫摩尔)在三氟乙酸(4mL)和二氯甲烷(4mL)的混合溶液，在16℃下搅拌1小时。LCMS显示反应完成。向混合物中加入碳酸钠水溶液(50mL)，并用二氯甲烷(50mL×2)萃取。有机层用无水硫酸钠干燥，过滤并浓缩，得到标题化合物(0.7克，粗品)为黄色固体。LCMS(ESI)(0-60AB):m/z:306.0[M+1].
实施例12D
3-(3-甲氧基-4-硝基苯基)-9-甲基-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000094
在16℃下，向实例12C(0.7克，2.3毫摩尔)的四氢呋喃(10mL)溶液中加入甲醛水溶液(207毫克，6.9毫摩尔，37％)，并在16℃下搅拌0.5小时。然后加入醋酸硼氢化钠(1.5克，6.9毫摩尔)，并在16℃下搅拌12小时。LCMS显示反应完成。将混合物用二氯甲烷(60mL)稀释，过滤并浓缩，得到标题化合物(0.6克，产率82％)为黄色油状物。LCMS(ESI)(0-60AB):m/z:320.2[M+1].
实施例12E
2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺
Figure PCTCN2015082605-appb-000095
向实施例12D(0.6克，1.9毫摩尔)的EtOH/H2O(12mL)混合液中加入铁粉(1.1克，18.8毫摩尔)和氯化铵(1.1克，18.8毫摩尔)。将混合物在80℃下搅拌2小时。TLC(DCM：甲醇＝6:1)显示反应已完成。将反应混合物过滤并浓缩。将得到的残余物通过制备TLC纯化(DCM：甲醇＝6:1)，得到标题化合物(400毫克，产率74％)为绿色固体。1H NMR(400MHz,CD3OD):δ,6.72(d,J＝8.4Hz,1H),6.64(d,J＝2.0Hz,1H),6.51(d,J＝8.4Hz,1H),3.86(s,3H),3.08-2.96(m,4H),2.58-2.45(m,4H),2.32(s,3H),1.75-1.52(m,8H).
实施例12F
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基硫醚
Figure PCTCN2015082605-appb-000096
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺，以及将(2-((2,5-二氯嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((2,5-二氯嘧啶-4-基)氨基)苯基)二甲基硫醚。得到标题化合物为棕色固体，产率36％。1H NMR(400MHz,CD3OD):δ,8.35(s,1H),7.89(dd,J＝13.6,8.0Hz,1H),7.75-7.80(m,2H),7.71(d,J＝8.8Hz,1H),7.57-7.66(m,2H),7.22(d,J＝8.4Hz,1H),4.02(s,3H),3.69(br.s.,4H),3.47(d,J＝12.8Hz,2H),3.18-3.29(m,2H),2.92(s,3H),2.19-2.38(m,2H),2.15-1.94(m,10H),1.75-1.91(m,2H).LCMS(ESI)(0-60AB):m/z:585.2[M+1].
流程C
Figure PCTCN2015082605-appb-000097
实施例13
(2-((5-氯-2-((4-(2-(二甲基氨基)-7-氮杂螺[3.5]壬-7-基)-2-甲氧基苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000098
实施例13A
叔丁基2-(二甲基氨基)-7-氮杂螺[3.5]壬烷-7-羧酸乙酯
Figure PCTCN2015082605-appb-000099
在室温下，N2保护下，向叔丁基-2-氧代-7-氮杂螺[3.5]壬烷-7-羧酸乙酯(200毫克，0.835毫摩尔)的MeOH(10mL)混合物中加入二甲胺盐酸盐(340.73毫克，4.18毫摩尔)和三乙胺(507.41毫克，5.01毫摩尔)。将反应混合物在室温下搅拌30分钟，然后加入醋酸硼氢化钠(531.38毫克，2.51毫摩尔)。将反应混合物在30℃下搅拌6小时。LCMS显示反应完成。将混合物减压浓缩，得到残余物，加水稀释(30mL)中并用DCM(50mL×2)萃取。将有机相用饱和盐水(50mL)洗涤，无水硫酸钠干燥，过滤并真空浓缩，得到标题化合物(200mg，产率89.16％)为白色固体。LCMS(ESI)(5-95AB):m/z:269.3[M+1].
实施例13B
N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺
Figure PCTCN2015082605-appb-000100
实施例13A(400.00毫克，1.49毫摩尔)的TFA(10mL)混合物，在30℃下搅拌12小时。TLC显示反应完成。将混合物减压浓缩，得到标题化合物(251毫克，粗品)为黄色油状物。
实施例13C
7-(3-甲氧基-4-硝基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺
Figure PCTCN2015082605-appb-000101
在氮气保护下，向实施例13B(251.00毫克，1.49毫摩尔)和4-氟-2-甲氧基-1-硝基苯(305.98毫克，1.79毫摩尔)的二甲基亚砜(10mL)的溶液中加入碳酸钾(617.80毫克，4.47毫摩尔)，将混合物在90℃下搅拌12小时。TLC显示反应完成。将混合物倒入水(25mL)中并用二氯甲烷(50mL×2)萃取。有机相用饱和盐水(50ml×2)洗涤，用无水硫酸钠干燥，过滤并浓缩，得到残余物，通过硅胶色谱纯化(二氯甲烷：甲醇＝80：1，60：1)，得到标题化合物(310毫克，产率65.14％)为黄色固体。LCMS(ESI)(5-95AB):m/z:320.3[M+1].
实施例13D
7-(4-氨基-3-甲氧基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺
Figure PCTCN2015082605-appb-000102
向实施例13C(310毫克，0.97毫摩尔)的甲醇(10mL)溶液中加入锌粉(317.33毫克，4.85毫摩尔)和氯化铵(207.66毫克，3.88毫摩尔)。将此悬浮液在30℃下搅拌0.5小时。TLC显示起始原料完全消耗。将反应混合物过滤，浓缩，得到标题化合物(185毫克，产率65.86％)为白色固体。
实施例13E
(2-((5-氯-2-((4-(2-(二甲基氨基)-7-氮杂螺[3.5]壬-7-基)-2-甲氧基苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000103
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为7-(4-氨基-3-甲氧基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺。得到标题化合物为白色固体，产率16％。1H NMR(400MHz,CD3OD):δ,8.29(s,1H),8.14(br.s,1H),7.68-7.82(m,3H),7.60(d,J＝2.0Hz,1H),7.54(t,J＝6.8Hz,1H),7.31(dd,J＝8.8,2.0Hz,1H),4.04(s,3H),3.85(t,J＝ 8.4Hz,1H),3.65-3.77(m,4H),2.86(s,6H),2.60(br.s.,2H),2.22-2.34(m,6H),1.85-1.92(m,6H).LCMS(ESI)(5-95AB):m/z:569.2[M+1].
实施例14
(2-((5-氯-2-((4-(9-(二甲基氨基)-3-氮杂螺[5.5]十一烷-3-基)-2-甲氧基苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000104
实施例14A
叔丁基10-(二甲基氨基)-3-氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000105
本实施例根据实施例13A的方法制备，将叔丁基2-氧代-7-氮杂螺[3.5]壬烷-7-羧酸乙酯替换为叔丁基-9-氧代-3-氮杂螺[5.5]十一烷-3-羧酸甲酯。得到标题化合物为黄色油状物，产率79.37％。
实施例14B
N，N-二甲基-3-氮杂螺[5.5]十一碳-9-胺
Figure PCTCN2015082605-appb-000106
本实施例根据实施例13B的方法制备，将叔丁基2-(二甲基氨基)-7-氮杂螺[3.5]壬烷-7-羧酸乙酯替换为叔丁基10-(二甲基氨基)-3-氮杂螺[5.5]十一烷-3-羧酸甲酯。得到标题化合物，为黄色油状物(粗品)，无需进一步纯化直接用于下一步骤。
实施例14C
3-(3-甲氧基-4-硝基苯基)-N，N-二甲基-3-氮杂螺[5.5]十一碳-9-胺
Figure PCTCN2015082605-appb-000107
本实施例根据实施例13C的方法制备，将N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺替换为N，N-二甲基-3-氮杂螺[5.5]十一碳-9-胺。得到标题化合物为深色固体，产率55％。LCMS(ESI)(5-95AB):m/z:348.1[M+1].
实施例14D
3-(4-氨基-3-甲氧基苯基)-N，N-二甲基-3-氮杂螺[5.5]十一碳-9-胺
Figure PCTCN2015082605-appb-000108
本实施例根据实施例13D的方法制备，将7-(3-甲氧基-4-硝基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺替换为3-(3-甲氧基-4-硝基苯基)-N，N-二甲基-3-氮杂螺[5.5]十一碳-9-胺。得到标题化合物为深色固体，产率82％。LCMS(ESI)(5-95AB):m/z:318.2[M+1].
实施例14E
(2-((5-氯-2-((4-(9-(二甲基氨基)-3-氮杂螺[5.5]十一烷-3-基)-2-甲氧基苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000109
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为3-(4-氨基-3-甲氧基苯基)-N，N-二甲基-3-氮杂螺[5.5]十一碳-9-胺。得到标题化合物为黄色固体，产率11％。1H NMR(400MHz,CD3OD):δ,8.29(s,1H),8.11-8.20(m,1H),7.70-7.80(m,3H),7.64(d,J＝2.8Hz,1H),7.52-7.56(m,1H),7.36(dd,J＝8.8,2.0Hz,1H),4.04(s,3H),3.58-3.86(m,4H),3.26-3.29(m,1H),2.91(s,6H),1.97-2.22(m 8H),1.91(s,3H),1.88(s,3H),1.73-1.82(m,2H),1.44-1.56(m,2H).LCMS(ESI)(0-60AB):m/z:597.3[M+1].
流程D
Figure PCTCN2015082605-appb-000110
实施例15
(2-((2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-5-(三氟甲基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000111
实施例15A
(2-((2-氯-5-(三氟甲基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000112
本实施例根据实施例1L的方法制备，将2,4,5-三氯嘧啶替换为2,4-二氯-5-(三氟甲基)嘧啶。得到标题化合物为白色固体，产率23％。LCMS(ESI)(5-95AB):m/z:350.0[M+1].
实施例15B
2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺与2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺
Figure PCTCN2015082605-appb-000113
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为2-甲氧基-4-(9-甲基-3-1,9-二氮杂螺[5.5]十一烷-3-基)苯胺，以及(2-((2,5-二氯 嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((2-氯-5-(三氟甲基)嘧啶-4-基)氨基)苯基)二甲基氧化膦。得到标题化合物为无色油收率20％。LCMS(ESI)(5-95AB):m/z:603.3[M+1].
实施例16
(2-((4-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-1,3,5-三嗪-2-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000114
实施例16A
(2-((4-氯-1,3,5-三嗪-2-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000115
向2,4-二氯-1,3,5-三嗪(50毫克，0.33毫摩尔)的MeCN(3.0mL)溶液中，加入DIPEA(52毫克，0.40毫摩尔)和(2-氨基苯基)二甲基氧膦(85毫克，0.50毫摩尔)。将反应混合物在20℃下搅拌2小时。TLC(DCM：甲醇＝20：1)显示反应完成。将混合物浓缩，得到残余物，用水(10mL)稀释，用HCl水溶液(1N)将溶液酸化以调节pH值至7，并用DCM(20mL×2)萃取。将有机层用无水硫酸钠干燥并浓缩，得到标题化合物(50mg，产率53.05％)为黄色油状物。LCMS(ESI)(0-30AB):m/z:282.9[M+1].
实施例16B
叔丁酯9-(4-((4-((2-(二甲基膦酰基)苯基)氨基)-1,3,5-三嗪-2-基)氨基)-3-甲氧基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000116
向实施例16A(40毫克，0.14毫摩尔)的MeCN(3.0mL)溶液中加入DIPEA(27毫克，0.21毫摩尔)和实施例8B(53毫克，0.14毫摩尔)。将该混合物在20℃下搅拌2小时。TLC(DCM：甲醇＝20：1)显示反应完成。将混合物浓缩，得到残余物，将其加水(10mL)稀释，将混合物用HCl水溶 液(1N)调节pH值至7，并用DCM(20mL×2)萃取。将有机层干燥并浓缩，得到标题化合物(50mg，产率56.8％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:622.3[M+1].
实施例16C
(2-((4-((2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-1,3,5-三嗪-2-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000117
实施例16B(50毫克，0.08毫摩尔)溶解在TFA(3.0mL)中，反应混合物在20℃下搅拌2小时。将溶液用二氯甲烷(5mL)中并浓缩至干，得到标题化合物(40mg，粗品)为黄色油状物。
实施例16D
(2-((4-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-1,3,5-三嗪-2-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000118
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((4-((2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-1,3,5-三嗪-2-基)氨基)苯基)二甲基氧膦。得到标题化合物为白色固体，产率15％。LCMS(ESI)(0-60AB):m/z:536.3[M+1].
实施例17
(2-((2-(氨基)-7H-吡咯并[2,3-D](2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)嘧啶唑-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000119
实施例17A
(2-((2-氯-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000120
在0℃下，向实施例1K(300毫克，1.77毫摩尔)的DMF(5.0mL)溶液中加入NaH(106毫克，2.66毫摩尔)，然后分批加入2,4-二氯-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶(677毫克，2.13毫摩尔)到反应混合物中，该反应混合物在20℃搅拌5小时。TLC(甲醇：DCM＝20：1)显示反应完成。将混合物用EtOAc(20mL)稀释，并用水洗涤(10mL)。将有机层干燥并浓缩。将粗产物通过制备TLC(DCM：甲醇＝10：1)纯化，得到标题化合物(270mg，产率33.8％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:451.2[M+1].
实施例17B
(2-((2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-7-((2-(三甲基甲硅烷基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000121
向实施例17A(187毫克，0.41毫摩尔)的二氧六环(5.0mL)溶液中加入实施例12E(100毫克，0.34毫摩尔)，Xphos(32毫克，0.07毫摩尔)，Pd2(dba)3(32毫克，0.03毫摩尔)和碳酸钾(95毫克，0.69毫摩尔)。将反应混合物在90℃搅拌16小时。TLC(PE：乙酸乙酯＝3:1)显示反应已完成。将反应悬浮液用乙酸乙酯(20mL)稀释并过滤。将有机层用水(50mL)洗涤并干燥，然后浓缩。将粗产物 通过制备TLC(DCM：甲醇＝10：1)纯化，得到标题化合物(100毫克，产率41.1％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:704.2[M+1].
实施例17C
(2-((2-(氨基)-7H-吡咯并[2,3-D](2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)嘧啶唑-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000122
实施例17B(100毫克，0.14毫摩尔)溶解在TFA(3.0mL)，反应混合物在20℃下搅拌30分钟。TLC(DCM：甲醇＝10：1)表明，起始原料消失。将反应混合物用DCM(10mL)稀释中并浓缩，得到粗品中间体(59毫克，粗品)；将该粗品中间体溶于甲醇(5.0mL)，然后加入NaOH调节pH值到12，并将该混合物在20℃搅拌30分钟。LCMS显示得到产品。将反应混合物用HCl水溶液(1N)中和，并通过制备HPLC纯化，得到标题化合物(10.58毫克，12.98％)为白色固体。LCMS(ESI)(0-60AB):m/z:574.3[M+1].
流程E
Figure PCTCN2015082605-appb-000123
实施例18
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)(甲基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000124
实施例18A
(2-((2,5-二氯嘧啶-4-基)(甲基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000125
向实施例1L(100毫克，0.32毫摩尔)，K2CO3(87.44毫克，0.63毫摩尔)的MeCN(2mL)溶液中加入MeI(0.1mL，1.58毫摩尔)。将反应混合物在70℃下搅拌8小时。TLC(DCM：甲醇＝20：1)显示反应完成。向该混合物中加入乙酸乙酯(50mL)并用盐水(30mL)洗涤，干燥并浓缩，得到黄色油状物。将该残余物通过制备TLC(DCM：甲醇＝20:1)纯化，得到标题化合物(50毫克，产率47.88％)为黄色油状物。1H NMR(400MHz,CDCl3):δ,8.05(s,1H),7.90-7.80(m,1H),7.60-7.48(m,2H),7.14-7.20(m,1H),3.55(s,3H),1.79(dd,J＝19.2,12.8Hz,6H).
实施例18B
(2-((5-氯-2-((2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)(甲基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000126
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为叔丁基3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯，以及将(2-((2,5-二氯嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((2,5-二氯嘧啶-4-基)(甲基)氨基)苯基)二甲基氧化膦。得到标题化合物为黄色油状物，产率的33％。LCMS(ESI)(0-60AB):m/z:569.2[M+1].
实施例18C
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)(甲基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000127
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4] 辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((5-氯-2-((2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)(甲基)氨基)苯基)二甲基氧膦。得到标题化合物为白色固体，产率32％。LCMS(ESI)(0-60AB):m/z:583.2[M+1].
实施例19
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氧)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000128
实施例12E(60毫克，0.2毫摩尔)，(2-((2,5-二氯嘧啶-4-基)氧基)苯基)二甲基氧膦(66毫克，0.2毫摩尔)，Xantphos(12毫克，0.02毫摩尔)，醋酸钯(5毫克，0.02毫摩尔)，碳酸铯(205毫克，0.6毫摩尔)的二氧六环(10mL)混合物，在N2保护下加热至100℃并搅拌12小时。LCMS显示反应完成。将反应混合物浓缩，并通过制备HPLC纯化，得到标题化合物(10.78毫克，收率9％)为白色固体。LCMS(ESI)(5-95AB):m/z:570.2[M+1].
流程F
Figure PCTCN2015082605-appb-000129
实施例20
(2-((5-氯-2-((5-氟-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000130
实施例20A
1-溴-2-氟-5-甲氧基-4-硝基苯
Figure PCTCN2015082605-appb-000131
向1-溴-2,5-二氟-4-硝基苯(5克，21.01毫摩尔)的甲醇(100mL)溶液中，加入甲醇钠(4.56克，84.43毫摩尔)。将混合液在60℃下搅拌12小时。TLC(石油醚：乙酸乙酯＝6:1)检测显示反应已完成。将反应液冷却至室温，减压浓缩。残留物用水(30mL)稀释，并用乙酸乙酯(50mL×2)萃取。有机相用饱和盐水(20mL×2)洗涤，用无水硫酸钠干燥，过滤浓缩。残留物用硅胶柱(石油醚：乙酸乙酯＝50：1，30：1，20：1)纯化，得到标题化合物(3.86克，15.44毫摩尔，73.48％产率)为黄色固体。1H NMR(400MHz,CDCl3):δ,7.72(d,J＝7.6Hz,1H),7.31(d,J＝5.6Hz),3.96(s,3H).
实施例20B
叔丁酯9-(2-氟-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000132
向实施例20A(120毫克，0.48毫摩尔)在二氧六环(10mL)的溶液中，加入叔丁基-3,9-二氮杂螺[5.5]十一烷-3-甲酸叔丁酯(139.59毫克，0.48毫摩尔)，Pd2(dba)3(43.95毫克，0.048毫摩尔)，Xantphos(22.85毫克，0.048毫摩尔)和碳酸铯(469.14毫克，1.44毫摩尔)；将反应混合物在100℃下搅拌12小时。TLC(PE：乙酸乙酯＝2:1)显示反应已完成。将混合物用EtOAc(20mL)稀释，依次用水(20mL)，盐水(20mL)洗涤，无水Na2SO4干燥，过滤并浓缩，得到黑色油状物，将其通过柱色谱(PE：乙酸乙酯＝30：1，20：1)分离纯化，得到标题化合物(148毫克，产率72.82％)为黄色固体。LCMS(ESI)(5-95AB):m/z:424.3[M+1].
实施例20C
叔丁酯9-(2-氟-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯
Figure PCTCN2015082605-appb-000133
本实施例根据实施例12C的方法制备，将叔丁酯9-(3-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸替换为叔丁酯9-(2-氟-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧 酸乙酯。得到标题化合物为黄色油状物(粗品)，将其直接用于下一步反应。
实施例20D
3-(2-氟-5-甲氧基-4-硝基苯基)-9-甲基-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000134
本实施例根据实施例12D的方法制备，将3-(3-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷替换为叔丁酯9-(2-氟-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸甲酯。得到标题化合物为黄色固体(粗品)，将其用于下一无需进一步纯化。LCMS(ESI)(5-95AB):m/z:338.2[M+1].
实施例20E
5-氟-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺
Figure PCTCN2015082605-appb-000135
在N2保护下，向实施例20D(90毫克，0.267毫摩尔)的MeOH(10mL)溶液中加入锌粉(87.21毫克，1.33毫摩尔)，氯化铵(57.07毫克，1.07毫摩尔)。反应混合物在25℃下搅拌0.5小时。TLC(DCM：甲醇＝10:1)显示原料完全消耗。将反应混合物过滤，浓缩，得到标题化合物(80mg，粗品)为黄色固体。
实施例20F
(2-((5-氯-2-((5-氟-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000136
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为5-氟-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一碳-3-基)苯胺。得到标题化合物为白 色固体，产率20％。1H NMR(400MHz,CD3OD)δ,8.34(s,1H),8.10(br.s.,1H),7.69-7.86(m,3H)7.51-7.59(m,1H),7.42(d,J＝6.8Hz,1H),4.02(s,3H),3.58-3.69(m,4H)3.47(d,J＝12.8Hz,2H),3.24(t,J＝12.8Hz,2H),2.93(s,3H),2.14-2.23(m,4H)1.96(br.s.,2H),1.90(d,J＝13.2Hz,6H),1.76-1.86(m,2H).LCMS(ESI)(5-95AB):m/z:587.3[M+1].
实施例21
(2-((2-((5-溴-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-5-氯嘧啶-4-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000137
实施例21A
1-溴-2-氟-4-甲氧基-5-硝基苯
Figure PCTCN2015082605-appb-000138
在0℃下，向1-溴-2-氟-4-甲氧基苯(5.00克，24.39毫摩尔)的硫酸(20.0mL)溶液中，分批加入硝酸钾(2.47克，24.39毫摩尔)，反应混合物在0℃搅拌0.5小时。TLC(石油醚：乙酸乙酯＝3:1)显示反应已完全。将反应混合物加入到冰水(50.0mL)中淬灭，并用EtOAc(100mL×2)萃取。有机层用无水硫酸钠干燥并过滤，减压浓缩至干，得到标题化合物(5.55克，产率90％)为白色固体。1H NMR(400MHz,CDCl3):δ,8.20(d.J＝6.8Hz,1H),6.91(d,J＝10Hz,1H),3.99(s,3H).
实施例21B
9-(2-溴-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000139
本实施例根据实施例4A的方法制备，将叔丁酯2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯替换为叔丁酯3,9-二氮杂螺[5.5]十一烷-3-甲酸叔丁酯，以及将4-氟-2-甲氧基-1-硝基苯替换为1-溴-2-氟-4-甲氧基-5-硝基苯。得到标题化合物为黄色油状物，产量为92％。LCMS(ESI)(0-60AB):m/z:484.2[M+1].
实施例21C
3-(2-溴-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000140
本实施例根据实施例12C的方法制备，将9-(3-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸替换为叔丁酯9-(2-溴-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯。得到标题化合物为黄色固体(粗品)，直接用于下一步反应。LCMS(ESI)(0-60AB):m/z:384.1[M+1].
实施例21D
3-(2-溴-5-甲氧基-4-硝基苯基)-9-甲基-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000141
本实施例根据实施例12D的方法制备，将3-(3-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷替换为3-(2-溴-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷。得到标题化合物为黄色油状物，产率为48％。LCMS(ESI)(0-60AB):m/z:398.1[M+1].
实施例21E
5-溴-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺
Figure PCTCN2015082605-appb-000142
本实施例根据实施例13D的方法制备，将7-(3-甲氧基-4-硝基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺替换为3-(2-溴5-甲氧基-4-硝基苯基)-9-甲基-3,9-二氮杂螺[5.5]十一烷。得到标题化合物为绿色固体，产率为72％。LCMS(ESI)(10-80CD):m/z:368.2[M+1].
实施例21F
(2-((2-((5-溴-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)-5-氯嘧啶-4-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000143
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为替换为5-溴-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺。得到标题化合物为无色油状物，产率为12％。LCMS(ESI)(5-95AB):m/z:649.1[M+1]
实施例22
(2-((5-氯-2-((5-氯-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000144
实施例22A
1-氯-2-氟-4-甲氧基苯
Figure PCTCN2015082605-appb-000145
向4-氯-3-氟苯酚(500毫克，3.41毫摩尔)的丙酮(10mL)溶液中加入碳酸钾(942.59毫克，6.82毫摩尔)和碘甲烷(1.94克，13.64毫摩尔)。将反应混合物在60℃搅拌16小时。TLC(石油醚：乙酸乙酯＝20：1)显示反应完成。将混合物浓缩以除去溶剂，向残余物加入水(20mL)并用乙酸乙酯(20mL×3)萃取。将有机层干燥并浓缩，得到标题化合物(470毫克，2.93毫摩尔，85.84％产率)为无色油状物。1H NMR(400MHz,CDCl3):δ,7.24-7.31(m,2H),6.71(dd,J＝10.8,2.8Hz,1H),6.65(dt,J＝8.8,1.4Hz,1H),3.80(s,3H).
实施例22B
1-氯-2-氟-4-甲氧基-5-硝基苯
Figure PCTCN2015082605-appb-000146
本实施例根据实施例21A的方法制备，将1-溴-2-氟-4-甲氧基苯替换为1-氯-2-氟-4-甲氧基 苯。得到标题化合物为黄色油状物，产率为51％。1H NMR(400MHz,CDCl3):δ,8.06(d,J＝7.6Hz,1H),6.92(d,J＝10.4Hz,1H),3.98(s,3H).
实施例22C
9-(2-氯-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000147
本实施例根据实施例4A的方法制备，将叔丁酯2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯替换为3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯，以及将4-氟-2-甲氧基-1-硝基苯替换为1-氯-2-氟-4-甲氧基-5-硝基苯。得到标题化合物为黄色油状物，产率为93％。1H NMR(400MHz,CDCl3):δ,8.04(s,1H),6.57(s,1H),3.96(s,3H),3.38-3.47(m,4H),3.16(br.s.,4H),1.69-1.73(m,4H),1.53(br.s.,4H),1.47(s,9H).
实施例22D
9-(4-氨基-2-氯-5-甲氧基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000148
本实施例根据实施例13D的方法制备，将7-(3-甲氧基-4-硝基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺替换为9-(2-氯-5-甲氧基-4-硝基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯。得到标题化合物为棕色油状物，产率为60％。LCMS(ESI)(10-80CD):m/z:310.2[M-100+1].
实施例22E
(2-((5-氯-2-((5-氯-2-甲氧基-4-(3,9-二氮杂[5.5]十一烷-3-基)苯基)氨基)吡啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000149
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为9-(4-氨基-2-氯-5-甲氧基苯基)-3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯。得到标题化合 物为棕色油状物，产率为35％。LCMS(ESI)(5-95AB):m/z:589.2[M+1].
实施例22F
(2-((5-氯-2-((5-氯-2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000150
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((5-氯-2-((5-氯-2-甲氧基-4-(3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)吡啶-4-基)氨基)苯基)二甲基氧膦。得到标题化合物为白色固体，产率为40％。1H NMR(400MHz,CDCl3):δ,8.35(s,1H),8.06(dd,J＝7.6,3.6Hz,1H),7.97(s,1H),7.80(dd,J＝13.2,7.2Hz,1H),7.73(t,J＝8.0Hz,1H),7.58(s,1H),7.50-7.57(m,1H),4.06(s,3H),3.74-3.78(m,4H),3.44-3.47(m,2H),3.24(t,J＝12.4Hz,2H),2.92(s,3H),2.15-2.30(m,4H),2.01(br.s.,2H),1.90(s,3H),1.87(s,3H),1.77-1.86(m,2H).LCMS(ESI)(5-95AB):m/z:603.2[M+1].
实施例23
(2-((5-氯-2-((2-甲氧基-5-甲基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000151
实施例23A
9-甲基-3,9-二氮杂螺[5.5]十一烷-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000152
在0℃下，向3,9-二氮杂螺[5.5]十一烷-3-甲酸叔丁酯(120毫克，0.47毫摩尔)的甲醇(5mL)溶液中加入37％甲醛溶液(71毫克，2.36毫摩尔)，反应混合物在16℃下搅拌0.5小时。然后将三乙酰氧基硼氢化钠(299毫克，1.42毫摩尔)加入到反应混合物中，并在16℃下搅拌12小时。LCMS显示反应完成。将混合物浓缩以除去溶剂，得到的残余物用水(10mL)稀释后用二氯甲烷(60mL)萃取，将有机层用无水硫酸钠干燥后过滤并浓缩，得到标题化合物为无色油状物(100毫克，产率为79％)。
实施例23B
3-甲基-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000153
实例23A(100毫克，0.37毫摩尔)在三氟乙酸(2mL)和二氯甲烷(2mL)的混合物溶液在16℃下搅拌1小时。LCMS显示反应完成。用碳酸钠溶液(50mL)将混合物碱化并用二氯甲烷(50mL×2)萃取。有机层用硫酸钠干燥，过滤并浓缩，得到标题化合物为棕色油状物(70mg)。
实施例23C
3-(5-甲氧基-2-甲基-4-硝基苯基)-9-甲基-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000154
本实施例根据实施例4A的方法制备，将叔丁酯2,7-二氮杂螺[3.5]壬烷-2-羧酸甲酯替换为3-甲基-3,9-二氮杂螺[5.5]十一烷，以及将4-氟-2-甲氧基-1-硝基苯替换为1-氟-5-甲氧基-2-甲基-4-硝基苯。得到标题化合物为黄色油状物，产率为40％。LCMS(ESI)(0-60AB):m/z:334.2[M+1].
实施例23D
2-甲氧基-5-甲基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺
Figure PCTCN2015082605-appb-000155
本实施例根据实施例13D的方法制备，将7-(3-甲氧基-4-硝基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺替换为3-(5-甲氧基-2-甲基-4-硝基苯基)-9-甲基-3,9-二氮杂螺[5.5]十一烷。得到标题化合物为棕色固体，产率为69％。LCMS(ESI)(10-80CD):m/z:304.3[M+1].
实施例23E
(2-((5-氯-2-((2-甲氧基-5-甲基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000156
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为2-甲氧基-5-甲基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺。得到标题化合物为白色固体，产率为19％。1H NMR(400MHz,CD3OD):δ,8.28(s,1H),8.17(br.s,1H),7.75-7.84(m,1H),7.62-7.74(m,2H),7.48-7.57(m,1H),7.42(s,1H),4.04(s,3H),3.56-3.84(m,4H),3.42-3.54(m,2H), 3.18-3.27(m,2H),2.94(s,3H),2.21-2.44(m,6H),2.01-2.13(m,2H),1.76-1.95(m,9H).LCMS(ESI)(5-95AB):m/z:583.3[M+1].
实施例24
(2-((5-氯-2-((4-甲氧基-6-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)吡啶-3-基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000157
实施例24A
3-(4-甲氧基-5-硝基吡啶-2-基)-9-甲基-3,9-二氮杂螺[5.5]十一烷
Figure PCTCN2015082605-appb-000158
在氮气保护下，将实施例23B(70毫克，0.42毫摩尔)，2-氯-4-甲氧基-5-硝基吡啶(94毫克，0.5毫摩尔)，Pd2(dba)3(38毫克，0.04毫摩尔)，Xantphos(24毫克，0.04毫摩尔)和碳酸铯(271毫克，0.83毫摩尔)二氧六环(5mL)的溶液，在90℃下搅拌12小时。LCMS显示反应完全。反应混合物加水(40mL)稀释后用二氯甲烷(50mL×2)萃取。将有机层用盐水(30mL×2)洗涤，无水硫酸钠干燥，过滤并浓缩。将所得残余物过硅胶柱纯化(石油醚：乙酸乙酯＝4：1，3：1)，得到标题化合物(50mg，产率为37.5％)为黄色油状物。
实施例24B
4-甲氧基-6-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)吡啶-3-胺
Figure PCTCN2015082605-appb-000159
本实施例根据实施例13D的方法制备，将7-(3-甲氧基-4-硝基苯基)-N，N-二甲基-7-氮杂螺[3.5]壬烷-2-胺替换为3-(4-甲氧基-5-硝基吡啶-2-基)-9-甲基-3,9-二氮杂螺[5.5]十一烷。得到标题化合物为棕色固体，产率为88％。
实施例24C
(2-((5-氯-2-((4-甲氧基-6-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)吡啶-3-基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000160
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为4-甲氧基-6-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)吡啶-3-胺。得到标题化合物为白色固体，产率为15％。LCMS(ESI)(10-80CD):m/z:570.2[M+1].
流程G
Figure PCTCN2015082605-appb-000161
实施例25
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)基-3-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000162
实施例25A
(2-氨基-3-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000163
在15℃下，向2-溴-6-氟-苯胺(1.00克，5.26毫摩尔)和二甲基氧膦(451.84毫克，5.79毫摩尔)在水(20mL)中的混合物中加入碳酸钾(2.91克，21.05毫摩尔)和钯/碳(150毫克)，反应混合物在 160℃微波加热下搅拌3小时。TLC(石油醚：乙酸乙酯＝10：1)显示反应完成。将反应混合物用二氯甲烷(20mL×4)萃取。将合并的有机层过滤后，将滤液浓缩并通过柱层析(二氯甲烷：甲醇＝1：0到10:1)分离得到标题化合物(100mg，产率为10.16％)为黄色固体。1H NMR(400MHz,CDCl3):7.12-7.03(m,1H),6.86(dd,J＝8.0,13.2Hz,1H),6.68-6.58(m,1H),5.52(br.s.,2H),1.79(d,J＝13.2Hz,6H).
实施例25B
(2-((2,5-二氯嘧啶-4-基)氨基)-3-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000164
在0℃下，向实施例25A(100毫克，0.534毫摩尔)的N，N-二甲基甲酰胺(2mL)的混合物中加入氢化钠(53.60毫克，1.34毫摩尔，60％)，然后将反应混合物在0℃搅拌0.5小时后，将2,4,5-三氯嘧啶(196毫克，1.07毫摩尔)加入到反应混合物中，反应混合物在15℃下搅拌16小时。LCMS显示反应完全。向该反应混合物中加入水(30mL)后并用乙酸乙酯(15mL×3)萃取。将合并的有机层用盐水(20mL)洗涤后浓缩，残留物通过柱层析(二氯甲烷：甲醇＝1：0到10:1)分离纯化，得到标题化合物(150毫克，产率为84.02％)为棕色油状物。1H NMR(400MHz,CDCl3):8.28(s,1H),7.53-7.33(m,3H),7.28-7.17(m,1H),5.54(br.s.,2H),1.85(d,J＝13.2Hz,6H).LCMS(ESI)(10-80CD):m/z:334.0[M+1].
实施例25C
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)基-3-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000165
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺，以及将(2-((2,5-二氯嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((2,5-二氯嘧啶-4基)氨基)-3-氟苯基)二甲基氧化膦。得到标题化合物为棕色固体，产率为17％。1H NMR(400MHz,CDCl3)：8.36(s，1H)，7.84-7.73(m,2H),7.71-7.62(m,2H),7.55-7.49(m,2H),7.16–7.09(m,1H),4.02(s,3H),3.80–3.60(m,4H),3.54–3.50(m,2H),3.29–3.20(m,2H),2.94(s,3H),2.47-1.90(m,6H),1.83(d,J＝13.6Hz,8H).LCMS(ESI)(0-60AB):m/z:587.3[M+1].
实施例26
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)基-5-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000166
实施例26A
(2-氨基-5-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000167
本实施例根据实施例1K的方法制备，将2-碘苯胺替换为4-氟-2-碘苯胺。得到标题化合物为棕色固体，产率为47％。1H NMR(400MHz,CDCl3):δ,6.96(m,1H),6.77(m,1H),6.60(m,1H),5.21(brs,2H),1.72-1.81(d,J＝22.8Hz,6H).LCMS(ESI)(10-80CD):m/z:188.1[M+1].
实施例26B
(2-((2,5-二氯嘧啶-4-基)氨基)-5-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000168
本实施例根据实施例1L的方法制备，将(2-氨基苯基)二甲基氧化膦替换为(2-氨基-5-氟苯基)二甲基氧化膦。得到标题化合物为白色固体，产率为50％。1H NMR(400MHz,CDCl3):δ,11.34(br.s.,1H),8.66(m,1H),8.19-8.28(m,1H),7.35-7.28(m,1H),6.98(m,1H)1.89-1.83(d,J＝13.2HZ,6H).LCMS(ESI)(5-95AB):m/z:334.1[M+1].
实施例26C
(2-((5-氯-2-((2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯基)氨基)嘧啶-4-基)氨基)基-5-氟苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000169
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺 替换为2-甲氧基-4-(9-甲基-3,9-二氮杂螺[5.5]十一烷-3-基)苯胺，以及将(2-((2,5-二氯嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((2,5-二氯嘧啶-4基)氨基)-5-氟苯基)二甲基氧化膦。得到标题化合物为白色固体，产率为10％。LCMS(ESI)(5-95AB):m/z:587.2[M+1].
流程H
Figure PCTCN2015082605-appb-000170
实施例27
((2-((5-氯-2-((2-甲氧基-4-(3-甲基-3-氮杂螺[5.5]十一碳-9-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000171
实施例27A
4-甲酰基哌啶-1-甲酸叔丁酯
Figure PCTCN2015082605-appb-000172
在-70℃，氮气保护下，向二甲基亚砜(4.37克，46.52毫摩尔)的二氯甲烷(25mL)溶液中滴加草酰氯(5.9克，46.52毫摩尔)的二氯甲烷(75mL)溶液；然后在-70℃下将4-(羟甲基)哌啶-1-甲酸叔丁酯(5克，23.26毫摩尔)的二氯甲烷(40mL)溶液逐滴加入到上述混合物中。反应混合物在-70℃下搅拌15分钟，逐滴加入三乙胺(11.76克，116.3毫摩尔)到反应混合物中。加完后反应混合物在-70℃下搅拌1小时，并升温至15℃。将反应混合物倒入水中，并用二氯甲烷(200mL)萃取。将有机层用碳酸氢钠溶液(100mL)洗涤后，无水硫酸钠干燥，过滤并浓缩至干。将所得油状物通过柱层析纯化(石油醚：乙酸乙酯＝10：1至3:1)，得到标题化合物(2.4克，产率为48％)为黄色油状物。1H NMR(400MHz,CDCl3):δ,9.68(s,1H),3.98-3.92(m,2H),2.96-2.91(m,2H),2.45-2.42(m,1H),1.89-1.60(m,2H),1.60-1.54(m,2H),1.47(s,9H).
实施例27B
9-氧代-3-氮杂螺[5.5]十一碳-7-烯-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000173
丁-3-烯-2-酮(0.658克，9.39毫摩尔)加入到实施例27A(2克，9.39毫摩尔)的四氢呋喃(100mL)溶液中。将反应混合物冷却至-5℃，将氢氧化钾-乙醇(3摩尔/升，1.57mL，4.7毫摩尔)溶液在5分钟内逐滴加入到反应混合物中。反应混合物升温至15℃并搅拌16小时。将石油醚(10mL)加入到反应混合物中，并将该混合物用盐水(100mL)洗涤。有机层浓缩，得到粗产物，粗产品通过硅胶柱层析纯化(石油醚：乙酸乙酯＝10：1至2:1)，得到标题化合物(1.12克，产率为45％)为黄色油状物。1H NMR(400MHz,CDCl3):δ,6.82(d,J＝10Hz,1H),5.97(d,J＝10Hz,1H),3.57-3.56(m,2H),2.50-2.47(m,2H),2.01-1.97(m,2H),1.67-1.65(m,2H),1.61-1.59(m,2H),1.49(s,9H).
实施例27C
9-氧代-3-氮杂螺[5.5]十一烷-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000174
向实施例27B(5.00克，18.84毫摩尔)的甲醇(100mL)溶液中加入钯/碳(200毫克，1.88毫摩尔)。将此悬浮液抽真空并用氢气置换几次。反应混合物在氢气(18psi)下保持温度10-25℃并搅拌5小时。将反应混合物过滤，滤液浓缩。粗产物通过硅胶柱层析纯化(石油醚：乙酸乙酯＝2：1)，得到标题化合物(4.78克，17.88毫摩尔，产率为94.9％)为白色固体。1H NMR(400MHz,CDCl3):δ,3.47-3.44(m,4H),2.38-2.35(m,4H),1.81-1.77(m,4H),1.58-1.56(m,4H),1.49(s,9H).
实施例27D
9-(((三氟甲基)磺酰)氧基)-3-氮杂螺[5.5]十一碳-8-烯-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000175
在-78℃氮气保护下，向实施例27C(2克，7.5毫摩尔)的四氢呋喃(20mL)溶液中逐滴加入二异丙基氨基锂(2.5M，1.22mL，9毫摩尔)，加完后，将反应混合物搅拌2小时。然后逐滴加入1,1,1-三氟-N-苯基-N-((三氟甲基)磺酰基)甲磺酰胺(2.67克，7.48毫摩尔，溶解在5mL的THF中)到反应液中，并在-78℃下搅拌1.5小时。将混合物升温至10℃搅拌2.5小时，反应混合物用氯化铵溶液(30mL)淬灭，并用乙酸乙酯(50mL×2)萃取。将有机层用盐水(50mL)洗涤，用Na2SO4干燥并浓缩，得到残余物，将其通过硅胶柱层析纯化(石油醚：乙酸乙酯＝5:1)，得到标题化合物(2.3g，产率为77％) 为棕色油状物。1H NMR(400MHz,CDCl3):δ,5.71(t,J＝4.0Hz,1H),3.51-3.45(m,2H),3.39-3.32(m,2H),2.37-2.35(m,2H),2.16-2.09(m,2H),1.71-1.67(m,2H),1.48(s,9H).1.45-1.42(m,4H).
实施例27E
4-溴-2-甲氧基苯胺
Figure PCTCN2015082605-appb-000176
向2-甲氧基苯胺(3克，24.36毫摩尔)的乙腈(30mL)溶液中加入N-溴代丁二酰亚胺(4.34克，24.36毫摩尔)，将反应混合物在15℃下搅拌15分钟。向反应混合物中加入亚硫酸钠溶液(40mL)淬灭反应，将混合物用乙酸乙酯(50ml)萃取；有机层用硫酸钠干燥，浓缩，得到粗产物，将其通过硅胶柱层析纯化(石油醚：乙酸乙酯＝10：1)得到标题化合物(2.7克，产率55％)为黄色固体。1H NMR(400MHz,CDCl3):δ,6.93-6.91(m,2H),6.62-6.59(dd,J＝1.6,2.0Hz,1H),3.86(s,1H).
实施例27F
2-甲氧基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)苯胺
Figure PCTCN2015082605-appb-000177
向实施例27E(500毫克，2.47毫摩尔)的DMSO(5mL)溶液中加入双联频那醇硼酸酯(628.4毫克，2.47毫摩尔)，四三苯基膦钯(150毫克，0.13毫摩尔)和醋酸钾(485毫克，4.95毫摩尔)，将反应混合物在150℃微波下搅拌40分钟。将反应混合物用乙酸乙酯(50mL)和水(30mL)稀释。有机层分离，干燥并浓缩，得到粗品，将其通过硅胶柱层析纯化(石油醚：乙酸乙酯＝10：1)得到标题化合物(160毫克，产率为26％)为黄色油状物。1H NMR(400MHz,CDCl3):δ,7.31(d,J＝8.0Hz,1H),7.22(s,1H),6.71(d,J＝8.0Hz,1H),3.91(s,3H),1.35(s,12H).
实施例27G
9-(4-氨基-3-甲氧基苯基)-3-氮杂螺[5.5]十一碳-8-烯-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000178
向实施例27D(300毫克，0.75毫摩尔)，实施例27F(187毫克，0.75毫摩尔)的二氧六环(10mL)溶液中加入Pd(dppf)Cl2(15毫克，0.075毫摩尔)和碳酸钾(204毫克，1.5毫摩尔)，将反应混合物在110℃搅拌16小时。将反应混合物通过硅藻土过滤，滤液浓缩，得到粗产物，将其通过硅胶柱层析纯化(石油醚：乙酸乙酯＝3:1)，得到标题化合物(110mg，产率为39％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:373.2[M+1].
实施例27H
9-(4-氨基-3-甲氧基苯基)-3-氮杂螺[5.5]十一烷-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000179
在N2保护下，向实施例27G(110毫克，0.3毫摩尔)的甲醇(5mL)溶液中加入钯/碳(10毫克，10％)，将反应混合物在氢气(15PSI)下，于16℃搅拌5小时。将反应混合物通过硅藻土过滤并真空浓缩，得到粗品标题化合物(130毫克)为黄色油状物。LCMS(ESI)(5-95AB):m/z:319.1[M-56+1].
实施例27I
(2-((5-氯-2-((2-甲氧基-4-(3-氮杂螺[5.5]十一碳-9-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000180
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为9-(4-氨基-3-甲氧基苯基)-3-氮杂螺[5.5]十一烷-3-羧酸叔丁酯。得到标题化合物为无色油状物，产率为15％。LCMS(ESI)(5-95AB):m/z:554.2[M+1].
实施例27J
((2-((5-氯-2-((2-甲氧基-4-(3-甲基-3-氮杂螺[5.5]十一碳-9-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000181
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((5-氯-2-((2-甲氧基-4-(3-氮 杂螺[5.5]十一烷-9-基)苯基)氨基)吡啶-4-基)氨基)苯基)二甲基氧膦。得到标题化合物为黄色固体，产率为27％。1H NMR(400MHz,CDCl3):δ,8.31-8.15(m,2H),7.75-7.70(m,1H),7.57(s,1H),7.45-7.37(m,2H),7.06(s,1H),6.91(s,1H),3.90(s,3H),3.44-3.36(m,2H),3.22-3.19(m,2H),2.91(s,3H),2.66-2.64(m 1H),2.44-1.93(m,2H),1.93(s,3H),1.92-1.88(d,J＝13.6Hz,6H),1.89(s,3H),1.84-1.60(m 9H),1.33-1.31(m,1H).LCMS(ESI)(5-95AB):m/z:568.2[M+1].
实施例28
(2-((5-氯-2-((2-(二氟甲氧基)-4-(3-甲基-3-氮杂螺[5.5]十一碳-9-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000182
实施例28A
5-氯-2-硝基-苯酚
Figure PCTCN2015082605-appb-000183
本实施例根据实施例11A的方法制备，将4-氟-2-甲氧基-1-硝基苯替换为4-氯-2-甲氧基-1-硝基苯。得到标题化合物为黄色油状物，产率为93％。1H NMR(400MHz,CDCl3):δ,10.67(s,2H),8.07(d,J＝8.8Hz,1H),7.20(d,J＝2.4Hz,1H),6.99(dd,J＝9.2,2.0Hz,1H).
实施例28B
4-氯-2-(二氟甲氧基)-1-硝基苯
Figure PCTCN2015082605-appb-000184
本实施例根据实施例11B的方法制备，将5-氟-2-硝基苯酚替换为5-氯-2-硝基苯酚，得到标题化合物为黄色油状物，产率为78％。1H NMR(400MHz,CDCl3):δ,7.92(d,J＝8.8Hz,1H),7.42-7.37(m,2H),6.64(t,J＝72.4Hz,1H).
实施例28C
2-(3-(二氟甲氧基)-4-硝基苯基)-4,4,5,5-四甲基-1,3,2-二氧硼戊环
Figure PCTCN2015082605-appb-000185
向实施例28C(223.56毫克，1.00毫摩尔)的二氧六环(10mL)溶液中加入双联频那醇硼酸酯(262毫克，1.00毫摩尔)，Pd(dppf)Cl2(73.17毫克，0.1毫摩尔)，三苯基膦(262.29毫克，1.00毫摩尔)和醋酸钾(196.28毫克，2.00毫摩尔)，将反应混合物换气并加热至100℃搅拌16小时。TLC(石油醚：乙酸乙酯＝10:1)显示原料完全消耗。将反应混合物倾入水(30mL)中并用乙酸乙酯(25mL×2)萃取。将有机相用盐水(30mL)洗涤，用无水硫酸镁干燥，真空浓缩，得到残余物，将其通过硅胶柱层析纯化(石油醚：乙酸乙酯＝30：1，20：1)，得到标题化合物(300毫克，产率为95.21％)为黄色固体。
实施例28D
9-(3-(二氟甲氧基)-4-硝基苯基)-3-氮杂螺[5.5]十一碳-8-烯-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000186
在氮气保护下，向实施例27D(948毫克，2.37毫摩尔)，实施例28C(500毫克，1.58毫摩尔)在二氧六环(4mL)和水(2mL)的混合液中加入Pd(dppf)Cl2(116毫克，0.16毫摩尔)和碳酸钠(503毫克，4.75毫摩尔)，将反应混合物在110℃下搅拌16小时。该反应混合物通过硅藻土过滤，浓缩，得到粗产品，将其通过硅胶柱层析纯化(石油醚：乙酸乙酯＝5:1)，得到标题化合物(602毫克，收率为58％)为黄色固体。1H NMR(400MHz,CDCl3):δ,7.92(d,J＝8.8Hz,1H),7.38-7.34(m,2H),6.82-6.28(m,2H),3.54-3.47(m,2H),3.38-3.33(m,2H),2.41(s,2H),2.18(d,J＝9.6Hz,1H),1.70-1.65(m,2H),1.46-1.42(m,5H).
实施例28E
9-(4-氨基-3-(二氟甲氧基)苯基)-3-氮杂螺[5.5]十一烷-3-羧酸叔丁酯
Figure PCTCN2015082605-appb-000187
向实施例28D(200毫克，0.46毫摩尔)的四氢呋喃(10mL)溶液中加入钯/碳(20毫克)；反应混合物在H2(50psi)于16℃搅拌5小时。将反应混合物通过硅藻土过滤并浓缩，得到标题化合物(150mg，产率为80％)为棕色固体。LCMS(ESI)(10-80CD):m/z:355.2[M+1-56].
实施例28F
(2-((5-氯-2-((2-(二氟甲氧基)-4-(3-氮杂螺[5.5]十一碳-9-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000188
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为9-(4-氨基-3-(二氟甲氧基)苯基)-3-氮杂螺[5.5]十一烷-3-甲酸叔丁酯。得到标题化合物为棕色油状物，产率为37.2％。
实施例28G
(2-((5-氯-2-((2-(二氟甲氧基)-4-(3-甲基-3-氮杂螺[5.5]十一碳-9-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000189
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4]辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((5-氯-2-((2-(二氟甲氧基)-4-(3-氮杂螺[5.5]十一碳-9-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦。得到标题化合物为白色固体，产率为24％。LCMS(ESI)(5-95AB):m/z:604.2[M+1].
实施例29
(2-((5-氯-2-((5-氟-2-甲氧基-4-(7-甲基-7-氮杂螺[3.5]壬烷-2-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦
Figure PCTCN2015082605-appb-000190
实施例29A
4-亚甲基哌啶-1-甲酸苄酯
Figure PCTCN2015082605-appb-000191
在室温N2保护下，向甲基(三苯基)溴化鏻(16.14克，45.17毫摩尔)和叔丁醇钾(5.58克，49.69毫摩尔)的四氢呋喃(350mL)混合物中一次性加入4-氧代哌啶-1-羧酸叔丁酯(9.00克，45.17毫摩尔)。反应混合物在16℃下搅拌3小时。TLC显示反应完成。将残余物倒入水中。水相用乙酸乙酯(200mL×3)萃取。将合并的有机相用饱和盐水(100mL)洗涤，无水硫酸钠干燥，过滤并真空浓缩。残余物通过硅胶柱层析纯化(石油醚：乙酸乙酯＝10：1，5：1)得到标题化合物(8.40克，产率为80.4％)为无色固体。1H NMR(400MHz,CDCl3):δ,7.41-7.34(m,5H),5.17(s 2H),4.78(s,2H),3.55-3.52(m,4H),2.24-2.21(m,4H).
实施例29B
1,1-二氯-2-氧代-7-氮杂螺[3.5]壬烷-7-羧酸苄酯
Figure PCTCN2015082605-appb-000192
在15-20℃下，向实施例29A(6.00克，25.94毫摩尔)和锌-铜(30.10克，233.46毫摩尔)的四氢呋喃(300mL)混合物中滴加2,2,2-三氯乙酰氯(23.58克，129.70毫摩尔)，滴加0.5小时后，将反应混合物在20-30℃下搅拌16小时。反应液缓慢的用冰淬灭，然后用乙酸乙酯(250mL×3)萃取。将合并的有机相用盐水(100mL×2)洗涤，无水硫酸钠干燥，过滤并真空浓缩。残余物通过硅胶柱层析纯化(石油醚：乙酸乙酯＝10：1，5：1)，得到标题化合物(3.81克，产率为42.81％)为白色固体。1H NMR(400MHz,CDCl3):δ,7.42-7.36(m,5H),5.17(s 2H),4.21-4.17(m,2H),3.13(s 2H),3.05-2.98(m,2H),2.02-1.96(m,2H),1.87-1.82(m,2H).
实施例29C
2-氧代-7-氮杂螺[3.5]壬烷-7-羧酸苄酯
Figure PCTCN2015082605-appb-000193
向实施例29B(3.00克，8.77毫摩尔)的甲醇(50mL)混合物中加入锌粉(1.72克，26.31毫摩尔)和氯化铵(4.69克，87.70毫摩尔)。反应混合物在10-20℃下搅拌0.5小时。将反应混合物倒入水(50mL)中，并搅拌20分钟。水相用乙酸乙酯(50mL×3)萃取。将合并的有机相用饱和食盐水(30mL)洗涤，无水硫酸钠干燥，过滤并真空浓缩。残余物通过硅胶柱层析纯化(石油醚：乙酸乙酯＝30：1，20：1)，得到标题化合物(2.12克，产率为88.44％)为黄色固体。1H NMR(400MHz,CDCl3):δ,7.40-7.35(m,5H),5.16(s 2H),3.54-3.51(m,4H),2.85(s,4H),1.76-1.74(m,4H).
实施例29D
2-(2-氟-5-甲氧基苯基)-2-羟基-7-氮杂螺[3.5]壬烷-7-羧酸苄酯
Figure PCTCN2015082605-appb-000194
在-60℃氮气保护下，向2-溴-1-氟-4-甲氧基-苯(400毫克，1.95毫摩尔)的四氢呋喃(15mL)混合物溶液中加入正丁基锂(249.83毫克，3.90毫摩尔)。将混合物在-60℃下搅拌1小时。然后在-60℃下将实施例29C(399.75毫克，1.46毫摩尔)用20分钟加入到反应混合物中。反应混合物在此温度下搅拌2小时，将反应混合物倒入水(50mL)中，并搅拌20分钟。水相用乙酸乙酯(50mL×3)萃取，合并的有机相用饱和盐水(50mL×2)洗涤，无水硫酸钠干燥，过滤并真空浓缩。残余物通过硅胶柱层析纯化(石油醚：乙酸乙酯＝10：1，1：1)，得到标题化合物(340毫克，产率为43.65％)为无色油状物。
实施例29E
2-(2-氟-5-甲氧基苯基)-7-氮杂螺[3.5]壬烷-7-羧酸苄酯
Figure PCTCN2015082605-appb-000195
在室温氮气保护下，向实施例29D(380毫克，0.951毫摩尔)的二氯甲烷(10mL)混合物中加入三乙基硅氢(221.24毫克，1.90毫摩尔)。反应混合物在18℃下搅拌1小时。将反应混合物倒入水(20mL)中，并搅拌20分钟。水相用乙酸乙酯(30mL×3)萃取。将合并的有机相用饱和食盐水(20mL)洗涤，无水硫酸钠干燥，过滤并真空浓缩。残余物通过硅胶柱层析纯化(石油醚：乙酸乙酯＝10：1，3：1)纯化，得到标题化合物(160毫克，产率为43.86％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:384.2[M+1].
实施例29F
2-(2-氟-5-甲氧基-4-硝基苯基)-7-氮杂螺[3.5]壬烷-7-羧酸苄酯
Figure PCTCN2015082605-appb-000196
在0-10℃下，向实施例29E(160毫克，0.417毫摩尔)的AcOH(4mL)混合物中，一次性加入硝酸(268.28毫克，4.17毫摩尔)。将反应混合物在18℃下搅拌2小时。将反应混合倒入冰水(20mL)中，并搅拌20分钟。水相用乙酸乙酯(40mL×3)萃取。将合并的有机相用饱和食盐水(20mL)洗涤， 无水硫酸钠干燥，过滤并真空浓缩。残余物通过硅胶柱层析纯化(石油醚：乙酸乙酯＝2：1)，得到标题化合物(145毫克，产率为81.10％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:451.1[M+Na].
实施例29G
5-氟-2-甲氧基-4-(7-氮杂螺[3.5]壬烷-2-基)苯胺
Figure PCTCN2015082605-appb-000197
在氮气保护下，向实施例29F(148.00毫克，345.43微摩尔)的四氢呋喃(10mL)混合物中加入钯/碳(40毫克，10％)。将反应液抽真空并用H2置换，将反应混合物在氢气(16psi)的条件下于10-25℃搅拌3小时。将反应混合物过滤，将滤液浓缩。粗产物通过硅胶柱层析纯化(二氯甲烷：甲醇＝20:1)得到标题化合物(50毫克，产率为54.76％)为黄色油状物。LCMS(ESI)(5-95AB):m/z:265.2[M+1].
实施例29H
(2-((5-氯-2-((5-氟-2-甲氧基-4-(7-氮杂螺[3.5]壬烷-2-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000198
本实施例根据实施例1M的方法制备，将2-甲氧基-5-(6-甲基-6-氮杂螺[3.4]辛-2-基)苯胺替换为5-氟-2-甲氧基-4-(7-氮杂螺[3.5]壬烷-2-基)苯胺。得到标题化合物为黄色油状物，产率为27.21％。LCMS(ESI)(5-95AB):m/z:544.2[M+1].
实施例29I
(2-((5-氯-2-((5-氟-2-甲氧基-4-(7甲基-7-氮杂螺[3.5]壬-2-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦
Figure PCTCN2015082605-appb-000199
本实施例根据实施例7的方法制备，将(2-((5-氯-2-((2-甲氧基-4-(2,6-二氮杂螺[3.4] 辛-6-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧化膦替换为(2-((5-氯-2-((5-氟-2-甲氧基-4-(7-氮杂螺[3.5]壬烷-2-基)苯基)氨基)嘧啶-4-基)氨基)苯基)二甲基氧膦。得到标题化合物为黄色固体，产率为42％。1H NMR(400MHz,CD3OD):δ,8.28(s,1H),8.25-8.19(m,1H),7.79-7.73(m,1H),7.63-7.61(m,1H),7.50-7.41(m,2H),6.98(d,J＝6.8,1H),3.93(s,3H),3.81-3.72(m,1H),3.52-3.39(m,2H),3.17-3.11(m,1H),3.01(m,1H),2.89(s,3H),2.56-2.51(m 1H),2.33-2.30(m,2H),2.19-2.06(m,2H),1.91-1.88(m,8H).LCMS(ESI)(5-95AB):m/z:558.2[M+1].
生化实验
实验材料
酶：ALK野生型，ALK C1156Y和ALK L1196M均购自Carna Biosciences(Japan)，EGFR T790M/L858R购自Life technology(Madison,WI)。
HTRF试剂盒：购自Cis-Bio International，内含Eu标记TK1抗体，XL665和biotin标记的TK1多肽底物。
检测仪器：Envision(PerkinElmer)。
实验方法
将测试化合物3倍梯度稀释，获得终浓度从1uM到0.017nM 11个剂量。
10ul野生型ALK酶反应混合物体系：0.5nM野生型ALK,1uM biotin-TK1peptide,30uM ATP。反应缓冲液：50mM Hepes(pH7.5),10mM MgCl2,0.01mM NaV3VO4.反应板为white Proxiplate 384-Plus plate(PerkinElmer)，室温反应90分钟。
10ul ALK C1156Y酶反应混合物体系:0.15nM ALK C1156Y,1uM biotin-TK1peptide,30uM ATP。反应缓冲液：50mM Hepes(pH7.5),10mM MgCl2,0.01mM NaV3VO4.反应板为white Proxiplate 384-Plus plate(PerkinElmer)。室温反应60分钟。
10ul ALK L1196M酶反应混合物体系:0.15nM ALK L1196M,1uM biotin-TK1peptide,30uM ATP。反应缓冲液：50mM Hepes(pH7.5),10mM MgCl2,0.01mM NaV3VO4.反应板为white Proxiplate 384-Plus plate(PerkinElmer)，室温反应60分钟。
10ul EGFR T790M/L858R酶反应混合物体系:0.08nM EGFR T790M/L858R,1uM biotin-TK1peptide,20uM ATP。反应缓冲液：50mM Hepes(pH7.5),10mM MgCl2,0.01mM NaV3VO4.反应板为white Proxiplate 384-Plus plate(PerkinElmer)。室温反应60分钟。
检测反应：加10ul检测试剂至反应板中，Antibody终浓度为2nM,XL665为62.5nM。室温孵育60分钟。Envision读板。
数据分析
通过下列公式将读数转化成抑制率(％)(Min-Ratio)/(Max-Min)*100％。4参数曲线拟合(Model 205in  XLFIT5,iDBS)测得IC50数据。
细胞实验
实验材料
RPMI1640，胎牛血清，青霉素/链霉素溶液，均购自Life Technology(Madison,WI)。Cell Titer-Glo luminescent cell viability reagents购自Promega(Madison,WI)。Karpas299cell line购自European Collection of Cell Cultures(ECACC)。读板仪器：Envision(PerkinElmer)。
实验方法
384孔板，每孔种2500个Karpas-299细胞，45ul体积。在CO 2培养箱中37℃过夜培养。待测化合物做3倍梯度稀释，获得浓度从2.5mM到0.127uM 10个剂量浓度，两复孔。中间板每孔加49ul培养基。从梯度稀释化合物板转移1ul化合物至中间板，混合充分。再从中间板取5ul液体转至细胞板。细胞继续在CO2培养箱中培养72小时。72小时后，加入25ul检测试剂。室温孵育10分钟，Envision读板。数据分析
通过下列公式将读数转化成抑制率(％)(Max-Sample)/(Max-Min)*100％。4参数曲线拟合(Model 205in XLFIT5,iDBS)测得IC 50数据。
本发明化合物的ALK酶抑制IC 50，ALK L1196M酶抑制IC 50，ALK C1156Y酶抑制IC 50，EGFR T790M/L858R酶抑制IC 50，以及Karpas-299细胞的ALK IC50的数据在以下表中展示。IC 50在1-100nM之间的化合物用+++表示；IC 50在101-1000nm之间的化合物用++表示，化合物的IC50大于1000nm化合物用+表示。
Figure PCTCN2015082605-appb-000200
Figure PCTCN2015082605-appb-000201
体内药效研究
以下体内药效数据表明，本发明的化合物在野生型LU-01-0015肺癌患者来源的异种移植(PDX)模型(BALB/c裸鼠)和LU-01-0319Crizotinib耐药模型(BALB/c裸鼠)这两种模型上都展示了比参考化合物AP26113意想不到的抗肿瘤活性以及减小肿瘤体积。举例来说，在LU-01-0015模型中，代表性化合物9，12，22，27等在给药(25毫克/公斤)23天后，肿瘤体积从最开始的约277mm3减小至30-45mm3，而AP26133只减少到119mm3。
1.在皮下植入LU-01-0015肺癌患者来源的异种移植(PDX)BALB/c裸小鼠上进行体内药效实验
BALB/c裸鼠，雌性，6-8周，体重约18-22克，将小鼠保持在一个特殊的无病原体的环境中，且在单个通风笼中(5只小鼠每笼)。所有的笼子，铺垫和水在使用前进行消毒。所有的动物都可以自由获取标准认证的商业实验室饮食。共有80只购于上海BK Laboratory Animal Co.,LTD的小鼠用于研究。每只小鼠在右胁腹皮下植入肿瘤组织(20-30立方毫米)，用于肿瘤的生长。当平均肿瘤体积达到约250-300立方毫米时开始实验。将试验化合物每日口服给药，25毫克/公斤。肿瘤体积每3天用二维卡尺测量，体积以立方毫米计量，通过以下公式计算：V＝V＝0.5a x b2，其中a和b分别是肿瘤的长径和短径。抗肿瘤药效是通过用化合物处理过的动物的平均肿瘤增加体积除以未处理过动物的平均肿瘤增加体积来确定。
Figure PCTCN2015082605-appb-000202
Figure PCTCN2015082605-appb-000203
2.在皮下移植LU-01-0319异种移植瘤，并对Crizotinib耐药BALB/c裸鼠上进行抗肿瘤体内药效实验
最初是从手术切除的临床样本中获得的LU-01-0319异种移植瘤模型，并植入裸鼠，此定义为批次P0(LU-01-0319-P0)。接下来的一批从P0的肿瘤植入定义为批次P1(LU-01-0319-P1)。FP3是从P2重新获得，接下来的批次从FP3肿瘤植入被定义为批次FP4。植入肿瘤后约2-3周，肿瘤大小达到约300mm3时，带有肿瘤小鼠用Crizotinib治疗。持续增加的肿瘤定义为LU-01-0319抗肿瘤模型。BALB/c裸鼠，雌性，6～8周，体重约18～22克，共75只小鼠用于研究，是从上海BK Laboratory Animal Co.,LTD购买。每只小鼠右翼皮下植入LU-01-0319R FP6的肿瘤切片(约30mm3)，用于肿瘤的生长。植入肿瘤后约2-3周，肿瘤大小达到约300mm3时，带有肿瘤的小鼠用Crizotinib(10/25/50/75毫克/千克)处理。根据肿瘤大小Crizotinib的剂量可以适当改变。当平均肿瘤体积达到约～500mm3时开始实验研究。试验化合物每天一次口服给药。肿瘤尺寸每周测量两次，用卡尺两个维度，并使用下面的公式计算体积：V＝0.5a x b2，其中a和b是分别肿瘤的长径和短径。抗肿瘤药效是通过用化合物处理过的动物的平均肿瘤增加体积除以未处理过动物的平均肿瘤增加体积来确定。
Figure PCTCN2015082605-appb-000204
*给药量3mg/Kg；**给药量10mg/Kg；***25mg/Kg
3.在皮下植入LU-01-0319肺癌患者来源的异种移植(PDX)BALB/c裸小鼠上进行体内药效实验
BALB/c裸鼠，雌性，6-8周，体重约18-22克，将小鼠保持在一个特殊的无病原体的环境中，且在单个通风笼中(5只小鼠每笼)。所有的笼子，铺垫和水在使用前进行消毒。所有的动物都可以自由获取标准认证的商业实验室饮食。共有80只购于上海BK Laboratory Animal Co.,LTD的小鼠用于研究。每只小鼠在右胁腹皮下植入肿瘤组织(20-30立方毫米)，用于肿瘤的生长。当平均肿瘤体积达到约250-300立方毫米时开始实验。将试验化合物每日口服给药，10毫克/公斤。肿瘤体积每3天用二维卡 尺测量，体积以立方毫米计量，通过以下公式计算：V＝V＝0.5a x b2，其中a和b分别是肿瘤的长径和短径。抗肿瘤药效是通过用化合物处理过的动物的平均肿瘤增加体积除以未处理过动物的平均肿瘤增加体积来确定。
Figure PCTCN2015082605-appb-000205
本发明的ALK抑制剂，可用于治疗各种癌症包括间变性大细胞淋巴瘤，非小细胞肺癌，扩散的大B-细胞淋巴瘤，炎性肌纤维母细胞肿瘤，成神经细胞瘤，甲状腺未分化癌和横纹肌肉瘤。ALK抑制剂，可以作为单独治疗或与其它化疗剂联合使用。

Claims (14)

  1. 式(I)所示化合物或其药学上可接受的盐，
    Figure PCTCN2015082605-appb-100001
    其中，
    T1选自N或C(R01)；
    T2选自-N(R01)-、O、S(＝O)2或-CH(NR01R02)-；
    R01或R02分别独立地选自H、或任选被1、2或3个卤素、羟基和/或氰基所取代的：C1-6烷基、C1-6杂烷基、C3-6环烷基-(CH2)0-3-和C3-6杂环烃基-(CH2)0-3-，其中所述“杂”代表1、2、或3个选自O、S、N、S(＝O)2和/或S(＝O)；
    任选地，T2上的R01和R02相互连接到同一个N上形成1个3～6元环，所述的环上含有1、2或3个杂原子，所述杂原子选自O、S和N；
    D1～D4分别独立地选自-(CR1R2)1-3-、O、S、C(＝O)、S(＝O)2和S(＝O)；
    D选自-N(R01)-、-O-和-S-；
    W选自＝O、＝S、＝N(CN)和＝N(OMe)；
    R3选自R03、OR03和SR03；
    R03选自C1-4烷基、C1-4卤代烷基和C3-5环烷基-(CH2)0-3-；
    Z选自N和C(R4)；
    X1选自C(Rx1)和N；
    X2选自C(Rx2)和N；
    Rx1、Rx2、R1、R2和R4分别独立地选自H、F、Cl、Br、I、CN、OH、SH、NH2、或任选被1、2或3个卤素、羟基和/或氰基所取代的：C1-6烷基、C1-6杂烷基、C3-6环烃基-(CH2)0-3-和C3-6杂环烃基-(CH2)0-3-，其中所述“杂”代表1、2、或3个杂原子，所述杂原子选自O、S和N；
    Rp1和Rp2分别独立地选自H、C1-4烷基和C1-4卤代烷基；
    任选地，Rp1和Rp2相互连接到同一个P上形成1个5～6元环，所述的环上含有1、2或3个杂原子，所述杂原子选自O、S、N和P；和
    任选地，Z和
    Figure PCTCN2015082605-appb-100002
    的位置可以互换。
  2. 根据权利要求1所述化合物或其药学上可接受的盐，其中所述R01和R02分别独立地选自H、CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3、-CH2CH2F、-CH2CH2S(＝O)2CH3、-CH2CH2CN、
    Figure PCTCN2015082605-appb-100003
    -CH2CH(OH)(CH3)2、-CH2CH(F)(CH3)2和-CH2CH2F。
  3. 根据权利要求1所述化合物或其药学上可接受的盐，其中所述R03选自CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3、-CH2CH2F和
    Figure PCTCN2015082605-appb-100004
  4. 根据权利要求1所述化合物或其药学上可接受的盐，其中所述Rp1和Rp2分别独立地选自H、CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3和-CH2CH2F。
  5. 根据权利要求1所述化合物或其药学上可接受的盐，其中所述Rx1、Rx2、R1、R2和R4分别独立地选自H、F、Cl、Br、I、CH3、CD3、CF3、CHF2、CH2CH3、CH(CH3)2、-CH2CF3、-CH2CH2CF3、-CH2CH2F、-CH2CH2S(＝O)2CH3、-CH2CH2CN、
    Figure PCTCN2015082605-appb-100005
    Figure PCTCN2015082605-appb-100006
    -CH2CH(OH)(CH3)2,-CH2CH(F)(CH3)2和-CH2CH2F。
  6. 根据权利要求1所述化合物或其药学上可接受的盐，其中T2上的NR01R02选自NHCH3、N(CH3)2、N(CH2CH3)2、
    Figure PCTCN2015082605-appb-100007
  7. 根据权利要求1所述化合物或其药学上可接受的盐，其中D、D1-4或T2分别独立地选自–NH-、-NMe-和-O-；D1-4和T2还可以选自-CH(NCH3CH3)-。
  8. 根据权利要求1所述化合物或其药学上可接受的盐，其中螺环结构单元
    Figure PCTCN2015082605-appb-100008
    选自
    Figure PCTCN2015082605-appb-100009
    Figure PCTCN2015082605-appb-100010
  9. 根据权利要求1所述化合物或其药学上可接受的盐，其具有式(Ⅱ)所示结构：
    Figure PCTCN2015082605-appb-100011
    其中各变量如权利要求1～8所定义。
  10. 根据权利要求1所述化合物或其药学上可接受的盐，其具有式(Ⅲ)所示结构：
    Figure PCTCN2015082605-appb-100012
    其中各变量如权利要求1～8所定义。
  11. 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：
    Figure PCTCN2015082605-appb-100013
    Figure PCTCN2015082605-appb-100014
  12. 根据权利要求1所述式(I)化合物的制备方法，其中T1代表N，T2代表NH，其制备路线如方案A或C所示：
    Figure PCTCN2015082605-appb-100015
    Figure PCTCN2015082605-appb-100016
    其中PG是氨基保护基，优选自BOC、Bn和Cbz，其他变量如上述权利要求所定义。
  13. 根据权利要求12所述化合物的制备方法，其中A5的制备方法如方案B所示：
    Figure PCTCN2015082605-appb-100017
  14. 根据权利要求1～11任意一项所述化合物或其药学上可接受的盐在制备治疗与ALK和/或EGFR以及它们的突变相关的癌症、与ROS1、BRAF、c-MET、HER2、KRAS/MEK、PIK3CA、FDFR、DDR2和/或VEGFR抑制剂联合治疗的癌症、或与细胞毒素联合治疗的癌症的药物中的应用。
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