WO2019223777A1 - 一种含有芳胺基取代的吡咯并嘧啶类化合物、制备方法及其应用 - Google Patents

一种含有芳胺基取代的吡咯并嘧啶类化合物、制备方法及其应用 Download PDF

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WO2019223777A1
WO2019223777A1 PCT/CN2019/088316 CN2019088316W WO2019223777A1 WO 2019223777 A1 WO2019223777 A1 WO 2019223777A1 CN 2019088316 W CN2019088316 W CN 2019088316W WO 2019223777 A1 WO2019223777 A1 WO 2019223777A1
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methyl
amino
ethyl
alkyl
propyl
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PCT/CN2019/088316
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English (en)
French (fr)
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张强
王中祥
冯守业
张宏波
杨海龙
周利凯
徐占强
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北京赛特明强医药科技有限公司
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Priority to CN201980017096.3A priority Critical patent/CN111836819A/zh
Publication of WO2019223777A1 publication Critical patent/WO2019223777A1/zh

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    • 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
    • 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 System
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the invention belongs to the technical field of medicine and relates to a pyrrolopyrimidine compound containing an arylamino group, a preparation method and an application thereof.
  • Protein kinase is an important signal messenger of cell life activity, which can catalyze the transfer of the ⁇ -phosphate group at the ATP terminus to the hydroxyl receptor in the amino acid residues of the substrate (serine, threonine, tyrosine), thereby activating the target Protein (Johnson LN, Lewis RJ, Structural basis for control by phosphorylation, Cheminform, 2001, 101,2209.). Protein kinases are involved in numerous physiological processes, including cell proliferation, survival, apoptosis, metabolism, transcription, and differentiation (Adams J.A., Kinetic and Catalytic Mechanisms of Protein Kinases, Chemical reviews, 2001, 101,2271.). Among the existing human drug targets, protein kinase family members account for up to 10% (Santos R., Ursu O., Gaulton A., et al., A comprehensive map of drug targets, Nature Reviews Drug Discovery, 2017, 16,19.).
  • Epidermal growth factor receptor (ErbB) tyrosine kinases can regulate cell proliferation, migration, differentiation, apoptosis, and cell migration through a variety of pathways. In various forms of malignant tumors, members of the ErbB family and their partial distributors are often overexpressed, amplified, or mutated, which makes them important therapeutic targets.
  • This family of protein kinases includes: ErbB1 / EGFR / HER1, ErbB2 / HER2, ErbB3 / HER3, and ErbB4 / HER4.
  • EGFR is an important target for the development of non-small cell lung cancer (dicamann, R., et.al., Personalizing, Therapy with Targeted Agents, Non-Small Cell, Cancer, ONCOTARGET, 2001, 2 (3), 165.).
  • Gefitinib, Erlotinib, and Icotinib are the first generation of EGFR-targeting reversible kinase inhibitors for the treatment of non-small cell cancer. These inhibitors have inhibitory effects on both wild-type and activated mutant EGFR, and have achieved great clinical success, but the resistance of the recipient patients after taking it for a period of time, especially the resistance caused by the T790M mutation Sex makes the effect less effective or ineffective.
  • Afatinib a second-generation EGFR inhibitor, is a non-reversible inhibitor that contains a Michael receptor and can covalently bond with a cysteine residue (Cys797) located at the entrance of the ATP-binding pocket.
  • the inhibitor shows strong activity against both T790M mutant EGFR kinase and wild-type EGFR kinase, and its inhibitory activity against T790M mutant EGFR kinase is higher than that of wild-type EGFR kinase, which makes the therapeutic window of the drug narrower in clinical application
  • the use effect is not ideal (Camidge, DR, et.al., Acquired, Resistance, TKIs, Solid Tumours: learning from cancer. Nature Reviews, Clinical Oncology, 2014, 11, 473.).
  • the third-generation EGFR kinase inhibitors Osimertinib and Olmutinib have achieved highly selective inhibition of T790M mutant EGFR kinase compared to wild-type EGFR kinase, widening the clinical use window, Effective treatment of patients with T790M mutation has been achieved.
  • three generations of EGFR kinase inhibitors have also developed resistance after a period of clinical use.
  • One of the reasons is the secondary mutation of C797S caused by EGFR.
  • Cruatinib is a targeted ALK inhibitor developed by Ariad Pharmaceuticals and approved by the US FDA in 2017 for the treatment of ALK-positive non-small cell lung cancer. According to the literature, Brigatinib has a certain inhibitory effect on C797S mutant EGFR kinase.
  • the present invention aims to provide an arylamine-substituted pyrrolopyrimidine compound, an isomer, a hydrate, a solvate, a pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutical combination thereof. And their use in the preparation of a medicament as a kinase inhibitor.
  • One aspect of the present invention provides a compound represented by formula (I), an isomer, a hydrate, a solvate, a pharmaceutically acceptable salt thereof, and a prodrug thereof,
  • X is CH or N
  • Y is CH or N
  • R 1 is -NHR 5
  • R 5 is unsubstituted or substituted aryl, heteroaryl, cycloalkyl, cycloalkyl bridged ring structure, and ring structure,
  • R 6 and R 7 are each independently -H, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl,
  • the parallel ring structure is selected from the group consisting of an aromatic ring and a 5-6 membered heteroaryl ring group, a 5-6 membered heteroaryl ring and a 5-6 member heteroaryl ring group, an aromatic ring and a 5-6 membered cycloalkyl group, and an aromatic ring 5-6 membered heterocyclic group, 5-6 membered heteroaryl ring and 5-6 membered cycloalkyl group or 5-6 membered heteroaryl ring and 5-6 membered heterocyclic group;
  • R 2 is -H, -CF 3 , -CH 2 CF 3 , C 1- C 6 alkyl, C 3- C 6 cycloalkyl, C 3- C 4 cycloalkyl substituted C 1- C 2 alkyl , A 4- to 6-membered heterocyclic group containing one oxygen atom, or-(CH 2 ) m R 8 , where m is an integer of 1, 2, 3,
  • R 3 and R 4 are each independently -H, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, 4-6 membered heterocyclic group containing one nitrogen atom or one oxygen atom, or-(CH 2 ) n R 9 , where n is an integer of 1, 2, 3,
  • the 4-6 membered heterocyclic ring containing one nitrogen atom or one oxygen atom is unsubstituted or substituted with a C 1 -C 3 alkyl group
  • R 3 , R 4 and the nitrogen atom to which it is attached form a 4-6 membered heterocyclic ring or a 6-9 membered spiro ring
  • the 4- to 6-membered heterocyclic ring formed by R 3 and R 4 and the nitrogen atom to which R 3 and R 4 are connected is unsubstituted or 1-2 members selected from halogen, cyano, hydroxyl, amino, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, halo C 1 -C 3 alkyl, cyano substituted C 1 -C 3 alkyl, hydroxy substituted C 1 -C 3 alkyl, C 1 -C 3 alkoxy Substituted with substituted C 1 -C 3 alkyl,
  • the 6-9 membered spiro ring formed by R 3 and R 4 and the nitrogen atom connected to it is a single spiro ring containing one nitrogen atom.
  • R 1 is -NHR 5 and R 5 is selected from the following groups:
  • T is NH, O or S
  • R 10 is selected from -H, -OH, -F, -Cl, -Br, -CN, -CF 3 , -OCF 3 , methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl , Methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclobutyloxy, or the following groups:
  • R 11 is -H, methyl, ethyl, propyl, isopropyl
  • R 12 is -H, -F, -Cl, -Br, hydroxyl, cyano, trifluoromethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, or Isopropoxy.
  • X is CH; Y is CH.
  • R 1 is
  • R 2 is selected from -H, -CF 3 , -CH 2 CF 3 , methyl, ethyl, propyl, isopropyl, 1-methylpropyl, 2-methylpropyl Base, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, propylene oxide-3-yl, tetrahydrofuran-3-yl, tetrahydropyridine Uran-4-yl, tetrahydropyran-3-yl, methylthioethyl, methylthiopropyl, methoxyethyl, methoxypropyl, ethoxyethyl, ethoxypropyl , Isopropoxyethyl, isopropoxypropyl, aminoacetyl, aminopropionyl, methanesulfony
  • R 2 is selected from n-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl.
  • R 3 and R 4 are each independently selected from the group consisting of -H, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, propylene oxide-3-yl, and tetrahydrofuran.
  • the substituted or unsubstituted 4-6 membered heterocyclic ring composed of the nitrogen atom to which R 3 and R 4 are connected is selected from the following ring structures:
  • R 13 is selected from -H, methylamino, ethylamino, dimethylamino,
  • R 14 is selected from -H, methyl, ethyl, propyl, isopropyl, formyl, acetyl or methanesulfonyl,
  • R 15 and R 16 are each independently selected from -H, -F, -CF 3 , hydroxyl, amino, cyano, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, and propoxy Methyl, isopropyloxy, cyanomethyl, cyanoethyl, methoxymethyl, methoxyethyl, methoxypropyl, hydroxymethyl, hydroxyethyl, hydroxypropyl,
  • the 6-9 membered spiro ring formed by R 3 and R 4 and the nitrogen atom to which it is connected is selected from the following spiro ring structures:
  • R 3 is selected from -H, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, iso Amyl, neopentyl, n-hexyl, isohexyl, cyclopropyl, cyclobutyl, cyclopentyl, propylene oxide-3-yl, tetrahydrofuran-3-yl, tetrahydropyran-4-yl, tetrahydro Pyran-3-yl, N-methylpiperidin-3-yl, N-methylpiperidin-4-yl, N-methylpyrrolidin-3-yl, N-methylazepine Alk-3-yl, methylthioethyl, methylthiopropyl, methoxyethyl, methoxypropyl, eth
  • R 4 is selected from n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl.
  • Another aspect of the present invention provides a compound represented by formula (I), an isomer, a hydrate, a solvate, a pharmaceutically acceptable salt thereof, and a prodrug thereof,
  • X is CH or N; Y is CH or N; preferably, X is CH; Y is CH;
  • R 1 is -NHR 5 and R 5 is selected from the following groups:
  • T is NH, O or S
  • R 10 is selected from -H, -OH, -F, -Cl, -Br, -CN, -CF 3 , -OCF 3 , methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl , Methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclobutyloxy, or the following groups:
  • R 11 is -H, methyl, ethyl, propyl, isopropyl
  • R 12 is -H, -F, -Cl, -Br, hydroxyl, cyano, trifluoromethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, or Isopropoxy
  • R 1 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • R 2 is selected from -H, -CF 3 , -CH 2 CF 3 , methyl, ethyl, propyl, isopropyl, 1-methylpropyl, 2-methylpropyl, cyclopropyl, cyclobutyl Group, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, oxetan-3-yl, tetrahydrofuran-3-yl, tetrahydropyran-4-yl, Tetrahydropyran-3-yl, methylthioethyl, methylthiopropyl, methoxyethyl, methoxypropyl, ethoxyethyl, ethoxypropyl, isopropoxyethyl Methyl, isopropyloxypropyl, aminoacetyl, aminopropionyl, methanesulfonylethy
  • R 3 is selected from -H, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl , Isohexyl, cyclopropyl, cyclobutyl, cyclopentyl, oxetan-3-yl, tetrahydrofuran-3-yl, tetrahydropyran-4-yl, tetrahydropyran-3-yl, N-methylpiperidin-3-yl, N-methylpiperidin-4-yl, N-methylpyrrolidin-3-yl, N-methylaza (hetero) cyclobutane-3-yl;
  • R 4 is selected from-(CH 2 ) n R 17 , where n is an integer of 1, 2, 3,
  • R 17 is Or C 3 -C 6 cycloalkyl, and R 18 is -OH, -CN, -C (O) NH 2 ,
  • Another aspect of the present invention provides a compound represented by formula (I), an isomer, a hydrate, a solvate, a pharmaceutically acceptable salt thereof, and a prodrug thereof,
  • X is CH or N
  • Y is CH or N
  • R 1 is -NHR 5 and R 5 is selected from the following groups:
  • T is NH, O or S
  • R 10 is selected from -H, -OH, -F, -Cl, -Br, -CN, -CF 3 , -OCF 3 , methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl , Methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclobutyloxy, or the following groups:
  • R 11 is -H, methyl, ethyl, propyl, isopropyl
  • R 12 is -H, -F, -Cl, -Br, hydroxyl, cyano, trifluoromethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, or Isopropoxy
  • R 2 is -H, C 1- C 6 alkyl, C 3- C 6 cycloalkyl, halo C 1- C 6 alkyl;
  • R 3 and R 4 are each independently -H, C 3 -C 8 cycloalkyl, or from 1 to 3 selected from C 1 -C 6 alkoxy, hydroxyl, halogen, C 3 -C 6 cycloalkyl C 1- C 9 alkyl substituted or substituted by
  • the 4- to 8-membered heterocyclic ring formed by R 3 and R 4 and the nitrogen atom to which they are connected is unsubstituted or 1-2 selected from halogen, cyano, hydroxyl, amino, C 1 -C 3 alkyl, C 1- C 3 alkoxy, halo C 1 -C 3 alkyl, cyano substituted C 1 -C 3 alkyl, hydroxy substituted C 1 -C 3 alkyl, C 1 -C 3 alkoxy substituted the C 1 -C 3 alkyl.
  • R 2 is -H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, isopentyl, pentyl, neopentyl, hexyl, isohexyl, cyclopropyl, Cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethyl, fluoroethyl;
  • R 3 and R 4 are each independently selected from -H, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, Neopentyl, n-hexyl, isohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetan-3-yl, tetrahydrofuran-3-yl, tetrahydropyran-4-yl, Tetrahydropyran-3-yl, N-methylpiperidin-3-yl, N-methylpiperidin-4-yl, N-methylpyrrolidin-3-yl, N-methyl nitrogen (hetero) Cyclobutane-3-yl, methylthioethyl, methylthiopropyl, methoxymethyl, methoxyethyl
  • the substituted or unsubstituted 4-8 membered heterocyclic ring composed of the nitrogen atom to which R 3 and R 4 are connected is selected from the following ring structures:
  • R 13 is selected from -H, methylamino, ethylamino, dimethylamino,
  • R 14 is selected from -H, methyl, ethyl, propyl, isopropyl, formyl, acetyl or methanesulfonyl,
  • R 15 and R 16 are each independently selected from -H, -F, -CF 3 , hydroxyl, amino, cyano, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, and propoxy Methyl, isopropyloxy, cyanomethyl, cyanoethyl, methoxymethyl, methoxyethyl, methoxypropyl, hydroxymethyl, hydroxyethyl, hydroxypropyl.
  • a method for preparing the compound, an isomer, a hydrate, a solvate, a pharmaceutically acceptable salt thereof, and a prodrug thereof including the following steps,
  • PG is selected from the group consisting of trimethylsilylethoxymethyl, tert-butoxycarbonyl, benzyloxycarbonyl, p-toluenesulfonyl, benzenesulfonyl, acetyl, trifluoroacetyl, fluorenylmethoxycarbonyl, benzyl; preferably trimethyl Silylethoxymethyl (SEM group),
  • R 1 , R 2 , R 3 , and R 4 are as described above.
  • the present application provides a pharmaceutical composition for treating a disease associated with tyrosine kinase EGFR, HER2, or ALK mutation or overexpression, which comprises the compound of formula (I) or a pharmaceutically acceptable compound thereof.
  • the salt or its hydrate or its solvate or its prodrug is composed of a pharmaceutically acceptable carrier or excipient.
  • a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, or prodrug thereof as an active ingredient, as described, One or more other therapeutic agents, and one or more pharmaceutically acceptable carriers or excipients.
  • the application also relates to the compound of formula (I), its isomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs in the preparation of treatments with tyrosine kinase EGFR, HER2, or ALK mutations or Application of an overexpression-related cancer and an autoimmune disease drug, wherein the cancer and autoimmune disease include fundus disease, dry eye, psoriasis, vitiligo, dermatitis, alopecia areata, rheumatoid arthritis, colitis, Multiple sclerosis, systemic lupus erythematosus, Crohn's disease, atherosclerosis, pulmonary fibrosis, liver fibrosis, bone marrow fibrosis, non-small cell lung cancer, small cell lung cancer, breast cancer, pancreatic cancer, glioma, Glioblastoma, ovarian cancer, cervical cancer, colorectal cancer, melanoma, endometrial cancer, prostate cancer, bladder cancer,
  • the application also relates to a method for treating a kinase-mediated disease or condition such as EGFR, HER2 or ALK, which comprises administering to a patient (human or other mammal, especially human) a therapeutically effective amount of the formula (I ) Compounds or salts thereof, said EGFR, HER2 or ALK-mediated diseases or conditions include those mentioned above.
  • substituted as referred to herein includes complex substituents (for example, aryl (such as phenyl), heteroalkyl, heteroaryl), more preferably 1 to 5 substituents, more preferably 1 There are three to three, preferably one to two, which can be freely selected from the substituent list.
  • complex substituents for example, aryl (such as phenyl), heteroalkyl, heteroaryl
  • substituents for example, aryl (such as phenyl), heteroalkyl, heteroaryl
  • substituents for example, aryl (such as phenyl), heteroalkyl, heteroaryl
  • alkyl refers to a saturated straight-chain, branched-chain hydrocarbon group having a specified number of carbon atoms
  • C 1 -C 10 alkyl refers to an alkyl moiety containing 1 to 10 carbon atoms
  • C 1 -C 3 Alkyl refers to an alkyl moiety containing 1 to 3 carbon atoms.
  • C 1 -C 6 alkyl includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl Base, tert-butyl, n-pentyl, 3- (2-methyl) butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl and 2-methyl Pentyl and others.
  • Alkoxy is an alkyl-O- group formed from a linear or branched alkyl group previously described with -O-, such as methoxy, ethoxy, and the like.
  • an alkylthio group is an alkyl-S- group formed from a linear or branched alkyl group previously described with -S-, such as methylthio, ethylthio, and the like.
  • Alkenyl and alkynyl include straight-chain, branched-chain alkenyl or alkynyl, and the term C 2 -C 6 alkenyl or C 2 -C 6 alkynyl means a straight-chain or branched hydrocarbon group having at least one alkenyl or alkynyl group.
  • Cycloalkyl means a non-aromatic, saturated, cyclic hydrocarbon group containing a specified number of carbon atoms.
  • (C3-C6) cycloalkyl refers to a non-aromatic cyclic hydrocarbon ring having 3-6 ring carbon atoms.
  • Exemplary "(C3-C6) cycloalkyl” includes cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.
  • aryl refers to an unsubstituted or substituted aromatic group, such as phenyl, naphthyl, anthracenyl.
  • aroyl refers to -C (O) -aryl.
  • heterocyclyl represents an unsubstituted or substituted stable 3 to 8 membered monocyclic saturated ring system, which is selected from carbon atoms and selected from N, O, and S It consists of 1 to 3 heteroatoms, of which N, S heteroatoms can be oxidized at will, and N heteroatoms can also be quaternized at will. Heterocyclic rings can be combined with any heteroatom or carbon atom to form a stable structure.
  • heterocycles include, but are not limited to, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, piperazinyl oxide, piperidinyl oxide, tetrahydrofuranyl, dioxolane Radicals, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiamorpholinyl sulfone and oxadiazolyl.
  • heteroaryl represents an unsubstituted or substituted stable 5- or 6-membered monocyclic aromatic ring system, and may also represent an unsubstituted or substituted 9 or 10-membered benzene benzene heteroaromatic ring system or bicyclic heteroaromatic ring system, which consists of carbon atoms and from 1 to 4 heteroatoms selected from N, O, S, of which N, S heteroatoms can be oxidized at will N heteroatoms can also be optionally quaternized.
  • Heteroaryl groups can adhere to any heteroatom or carbon atom to form a stable structure.
  • heteroaryl groups include, but are not limited to, thiathranyl, furyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl , Pyridyl, pyridazinyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuranyl, benzothienyl, benzoisoxazolyl, benzoxazolyl, Benzopyrazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, adenine, quinolinyl or isoquinolinyl.
  • carbonyl refers to a C (O) group.
  • alkyl or “aryl” or any of their prefixes appears in the name of a substituent (eg, aralkyl, dialkylamino), it will be considered to include the above as “alkane” And “aryl”.
  • a specified number of carbon atoms (for example, C1-C6) will independently represent the number of carbon atoms in an alkyl moiety or an alkyl moiety in which a alkyl group is used as its prefix stem.
  • solvated form may be a water-soluble form.
  • the invention includes all of these solvated and unsolvated forms.
  • the compounds of the present invention may have asymmetric carbon atoms. Based on their physical and chemical differences, such diastereomeric mixtures can be separated by known methods, such as chromatography or fractional crystallization. Into a single diastereomer. Enantiomers can be separated by first reacting with an appropriate optically active compound to convert the enantiomeric mixture into a diastereomeric mixture, separating the diastereomers, and then converting the single diastereomer Enantiomers are converted (hydrolyzed) to the corresponding pure enantiomers. All such isomers, including diastereomeric mixtures and pure enantiomers, are considered to be part of the invention.
  • the compound of the present invention as an active ingredient, and a method for preparing the same are the contents of the present invention.
  • the crystalline forms of some compounds can exist as polycrystals, and this form can also be included in the current invention.
  • some compounds can form solvates with water (ie, hydrates) or common organic solvents, and such solvates are also included in the scope of this invention.
  • the compounds of the present invention can be used for treatment in free form or, where appropriate, in the form of a pharmaceutically acceptable salt or other derivative.
  • pharmaceutically acceptable salt refers to the organic and inorganic salts of the compounds of the present invention. This salt is suitable for humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and is reasonable. Benefit / risk ratio.
  • Pharmaceutically acceptable salts of amines, carboxylic acids, phosphonates, and other types of compounds are well known in the art.
  • the salt can be formed by reacting a compound of the present invention with a suitable free base or acid.
  • salts with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, Alternatively, these salts can be obtained by using methods well known in the art, such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, besylate, benzoate, bisulfate, borate, butyrate, and camphoric acid Salt, camphor sulfonate, citrate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glyceryl phosphate, gluconic acid Salt, hemisulfate, hexanoate, hydroiodate, 2-hydroxyethanesulfonate, lactate, lactate, laurate, lauryl sulfate, malate, maleate, methane Sulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectate, persulfate, per-3-phenylpropionate, Phosphate, picrate, propionate
  • alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Other pharmaceutically acceptable salts include appropriate non-toxic ammonium, quaternary ammonium, and use such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkylsulfonate and arylsulfonate Amine cations.
  • prodrug as used herein means that a compound can be converted into a compound represented by formula (I) of the present invention in vivo. This conversion is affected by the hydrolysis of the prodrug in the blood or the enzymatic conversion into the parent compound in the blood or tissue.
  • the pharmaceutical composition of the present invention comprises a compound of formula (I) described herein or a pharmaceutically acceptable salt thereof, a kinase inhibitor (small molecule, polypeptide, antibody, etc.), an immunosuppressive agent, an anticancer drug, an antiviral agent, an antiviral agent Inflammatory agents, antifungal agents, antibiotics or additional active agents of anti-hyperplasia compounds; and any pharmaceutically acceptable carrier, adjuvant or excipient.
  • a kinase inhibitor small molecule, polypeptide, antibody, etc.
  • an immunosuppressive agent an anticancer drug
  • an antiviral agent an antiviral agent
  • Inflammatory agents antifungal agents
  • antibiotics or additional active agents of anti-hyperplasia compounds any pharmaceutically acceptable carrier, adjuvant or excipient.
  • the compounds of the invention can be used alone or in combination with one or more other compounds of the invention or with one or more other agents.
  • the therapeutic agents can be formulated to be administered simultaneously or sequentially at different times, or the therapeutic agents can be administered as a single composition.
  • the so-called "combination therapy" refers to the use of the compound of the present invention together with another agent.
  • the mode of administration is the simultaneous administration of each agent or the sequential administration of each agent. In either case, the purpose is to To achieve the best results of the drug.
  • Co-administration includes simultaneous delivery of the dosage forms, as well as separate dosage forms for each compound.
  • the administration of the compound of the present invention can be used concurrently with other known therapies in the art, for example, the use of radiation therapy or additional therapies such as cytostatic agents, cytotoxic agents, other anticancer agents in cancer treatment to improve Symptoms of cancer.
  • additional therapies such as cytostatic agents, cytotoxic agents, other anticancer agents in cancer treatment to improve Symptoms of cancer.
  • the invention is not limited to the order of administration; the compounds of the invention may be administered previously, concomitantly, or after other anticancer or cytotoxic agents.
  • one or more compounds or salts of formula (I) as its active ingredient can be intimately mixed with the pharmaceutical carrier, which is performed according to the traditional pharmaceutical ingredient technology,
  • the carrier can adopt various forms according to the preparation form designed according to different administration modes (for example, oral or parenteral administration).
  • Suitable pharmaceutically acceptable carriers are well known in the art. A description of some of these pharmaceutically acceptable carriers can be found in the Handbook of Pharmaceutical Excipients, a book jointly published by the American Pharmaceutical Association and the British Pharmaceutical Society.
  • the pharmaceutical composition of the present invention may have the following forms, for example, suitable for oral administration, such as tablets, capsules, pills, powders, sustained release forms, solutions or suspensions; for parenteral injections such as clear liquids, suspensions, Emulsions; or for topical application such as creams, creams; or as suppositories for rectal administration.
  • Pharmaceutical ingredients may also be suitable for precise single-dose administration in unit dosage form.
  • the pharmaceutical ingredient will include a traditional pharmaceutical carrier or excipient and a compound made as an active ingredient according to the current invention, and may also include other medical or pharmaceutical preparations, carriers, adjuvants, and the like.
  • Therapeutic compounds can also be administered to mammals rather than humans.
  • the dosage of a drug administered to a mammal will depend on the species of the animal and its disease status or its disorder.
  • the therapeutic compound can be administered to animals in the form of capsules, boluses, or tablet potions.
  • Therapeutic compounds can also be introduced into animals by injection or infusion. We prepare these medicinal forms in a traditional way that meets the standards of veterinary practice.
  • pharmaceutical synthetic drugs can be mixed with animal feed and fed to animals. Therefore, concentrated feed additives or premixes can be prepared to mix ordinary animal feed.
  • Yet another object of the present invention is to provide a method for treating cancer in a subject in need, comprising administering to the subject a method of treating a therapeutically effective amount of a composition containing a compound of the present invention.
  • the present invention also includes the use of the compound of the present invention or a pharmaceutically acceptable derivative thereof for the preparation of a medicament for treating cancers related to tyrosine kinases EGFR, HER2, ALK and autoimmune diseases.
  • the cancers including non-solid tumors, solid tumors, primary or metastatic cancers, as indicated elsewhere herein and including one or more other treatments that are resistant or refractory to the cancer
  • other diseases including but It is not limited to agents for fundus diseases, psoriasis, atherosclerosis, pulmonary fibrosis, liver fibrosis, bone marrow fibrosis, etc.).
  • the cancer includes, but is not limited to, non-small cell lung cancer, small cell lung cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, ovarian cancer, cervical cancer, colorectal cancer, melanoma, intrauterine Membrane cancer, prostate cancer, bladder cancer, leukemia, gastric cancer, liver cancer, gastrointestinal stromal tumor, thyroid cancer, chronic myelogenous leukemia, acute myeloid leukemia, non-Hodgkin lymphoma, nasopharyngeal cancer, esophageal cancer, brain Tumor, B-cell and T-cell lymphoma, lymphoma, multiple myeloma, biliary sarcoma, and bile duct cancer.
  • the present invention also provides a method for preparing the corresponding compound.
  • a variety of synthetic methods can be used to prepare the compounds described herein, including the following methods.
  • the compound of the present invention or a pharmaceutically acceptable salt, isomer or hydrate thereof may be The following methods are used to synthesize synthetic methods known in the field of organic chemical synthesis, or by those skilled in the art to understand variations of these methods. Preferred methods include, but are not limited to, the following methods.
  • a method for preparing a compound of the present invention, an isomer, a hydrate, a solvate, a pharmaceutically acceptable salt thereof, and a prodrug thereof, comprising the following steps:
  • R 1 , R 2 , R 3 , and R 4 are as defined above.
  • step 1) The compound represented by formula (IV) in step 1) and the compound represented by formula (III) are sufficiently contacted to obtain the compound represented by formula (II);
  • the reaction can be performed in an organic solvent, including, but not limited to, N, N-dimethylformamide (DMF), N, N-dimethylacetamide (DMA), N- One or a combination of two or more of methyl-2-pyrrolidone (NMP), ethylene glycol dimethyl ether, isopropanol, n-butanol, 2-butanol (sec-butanol), and tert-butanol;
  • NMF N, N-dimethylformamide
  • DMA N-dimethylacetamide
  • NMP N- One or a combination of two or more of methyl-2-pyrrolidone
  • ethylene glycol dimethyl ether isopropanol, n-butanol, 2-butanol (sec-butanol), and tert-butanol
  • NMP N-dimethyl-2-pyrrolidone
  • the reaction can be performed in the presence of a base, which includes but is not limited to: potassium carbonate, sodium carbonate, sodium acetate, triethylamine, diisopropylethylamine, triethylenediamine, pyridine, One or a combination of two or more of 4-dimethylaminopyridine, 1,8-diazabicycloundec-7-ene or N-methylmorpholine;
  • a base which includes but is not limited to: potassium carbonate, sodium carbonate, sodium acetate, triethylamine, diisopropylethylamine, triethylenediamine, pyridine, One or a combination of two or more of 4-dimethylaminopyridine, 1,8-diazabicycloundec-7-ene or N-methylmorpholine;
  • the reaction can be performed in the presence of an acid, which includes but is not limited to: trifluoroacetic acid, p-toluenesulfonic acid;
  • the reaction can be performed under the conditions of a palladium metal-catalyzed coupling reaction condition, wherein the palladium metal-catalyzed coupling reaction conditions are the palladium ligands, solvents, and bases used in common Buchward-Hartwig reactions;
  • Step 2 The compound represented by formula (II) in the presence of tetrabutylammonium fluoride is used to obtain a compound represented by formula (I).
  • Step 1) (2-((2-((4- (4- (dimethylamino) piperidin-1-yl) -3-methoxyphenyl) amino) -7-((2- (trimethyl Silyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 2) (2-((2-((4- (4- (dimethylamino) piperidin-1-yl) -3-methoxyphenyl) amino) -7H-pyrrolo [2,3- d) Preparation of pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 4) and Step 5) (2-((2-((4- (4- (dimethylamino) piperidin-1-yl) -3-isobutoxyphenyl) amino) -7H pyrrolo Preparation of [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Reference Example 1 was prepared in steps 1) and 2).
  • the starting material was equimolar equivalent of 1- (4-amino-2-isobutoxyphenyl) -N, N-dimethylpiperidine- 4-amine replaces 1- (4-amino-2-methoxyphenyl) -N, N-dimethylpiperidine-4-amine.
  • Example 7 For the preparation of Example 7, refer to Step 1) to Step 5) of Example 6, and replace the 1-iodo-2-methylpropane with an equimolar equivalent of iodocyclobutane in the starting material in step 1).
  • Example 8 refers to steps 1) to 5) of the preparation route of Reference Example 6, wherein the starting material in step 1) is replaced by 1-iodo-2-methylpropane with an equimolar equivalent of methyl iodide, and step 2 ) In place of N, N-dimethylpiperazin-4-amine with equimolar equivalent of 1-methyl-4- (piperidin-4-yl) piperazine.
  • Example 9 For the preparation of Example 9, refer to steps 1) to 5) of the preparation route of Example 6, wherein in step 2), equimolar equivalent of 1-methyl-4- (piperidin-4-yl) piperazine was used instead of N. , N-dimethylpiperazine-4-amine.
  • Example 10 For the preparation of Example 10, refer to steps 1) to 5) of the preparation route of Example 7, wherein in step 2), the molar equivalent of 1-methyl-4- (piperidin-4-yl) piperazine was used instead of N. , N-dimethylpiperazine-4-amine.
  • step 1) equimolar equivalent of bromoisopropane is used instead of 1-iodo-2-methylpropane
  • step 2) The equivalent molar equivalent of 2-methyl-1- (methyl (piperidin-4-yl) amino) propan-2-ol was used in place of N, N-dimethylpiperazin-4-amine.
  • step 1) equimolar equivalent of bromoisopropane is used instead of 1-iodo-2-methylpropane
  • step 2) In the equimolar equivalent of N- (2-methoxyethyl) -N-methylpiperidine-4-amine, N, N-dimethylpiperazine-4-amine was replaced.
  • Example 13 For the preparation of Example 13, refer to steps 1) to 5) of the preparation route of Example 6, wherein in step 1), equimolar equivalent of bromoisopropane is used instead of 1-iodo-2-methylpropane, and step 2) In the equimolar equivalent of N- (cyclopropylmethyl) -N-methylpiperidine-4-amine, N, N-dimethylpiperazine-4-amine was replaced.
  • Example 14 For the preparation of Example 14, refer to steps 1) to 5) of the preparation route of Example 6, wherein in step 2), an equimolar equivalent of 1-isopropyl-4- (piperidin-4-yl) piperazine was substituted. N, N-dimethylpiperazin-4-amine.
  • Step 7) (2-((2-((3-ethoxy-4- (4-((2-hydroxy-2-methylpropyl) (methyl) amino) piperidin-1-yl) benzene ) Amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Example 18 For the preparation method of Example 18, refer to the preparation steps 1) to 7) of Example 17, wherein in step 2), equimolar equivalent of 1-bromopropane is used instead of bromoethane.
  • Example 19 For the preparation method of Example 19, refer to the preparation steps 1) to 7) of Example 17, wherein in step 2), an equimolar equivalent of 1-bromo-2-methylpropane is used instead of bromoethane.
  • Example 20 For the preparation method of Example 20, refer to the preparation steps 1) to 7) of Example 17, wherein in step 2), equimolar equivalent of 1-bromobutane is used instead of bromoethane.
  • Example 21 For the preparation method of Example 21, refer to the preparation steps 1) to 7) of Example 17, wherein in step 2), equimolar equivalent of 1-bromo-3-methylbutane is used instead of bromoethane.
  • Example 28 For the preparation method of Example 28, refer to the preparation steps 1) to 7) of Example 17, wherein in step 2), an equimolar equivalent of bromocyclobutane is used instead of bromoethane.
  • Example 30 For the preparation method of Example 30, refer to the preparation steps 1) to 7) of Example 17, wherein in step 2), an equimolar equivalent of bromocyclopentane is used instead of bromoethane.
  • Step 3) to Step 5) (2-((2-((4- (4- (ethyl (2-hydroxy-2-methylpropyl) amino) piperidin-1-yl) -3-isopropyl Preparation of oxyphenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 3) to Step 5) Referring to Step 5) to Step 7) in the preparation method of Example 17, the starting material is equimolar equivalent of 1- (ethyl (1- (2-isopropoxy-4- Nitrophenyl) piperidin-4-yl) amino) -2-methylpropan-2-ol instead of 1-((1- (2-ethoxy-4-nitrophenyl) piperidine-4- (Methyl) (methyl) amino) -2-methylpropan-2-ol.
  • Example 32 For the preparation method of Example 32, refer to the preparation steps 1) to 7) of Example 17, wherein in step 2), an equimolar equivalent of bromocyclohexane is used instead of bromoethane.
  • Step 2) to Step 4) (2-((2-((4- (4- (azacyclooctane-1-yl) piperidin-1-yl) -3-isopropoxyphenyl) amino ) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 2) to Step 4) Referring to Step 5) to Step 7) in the preparation method of Example 17, the starting material is equimolar equivalent of 1- (1- (2-isopropoxy-4-nitrobenzene) (Yl) piperidin-4-yl) azacyclooctane instead of 1-((1- (2-ethoxy-4-nitrophenyl) piperidin-4-yl) (methyl) amino) -2 -Methylpropan-2-ol.
  • Step 7) (2-((2-((4- (4-(((1-hydroxycyclopropyl) methyl) (methyl) amino) piperidin-1-yl) -3-isopropoxy Phenyl) amino) -7-((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) di
  • step 6 For the preparation method, refer to step 6) in Example 17, wherein in an equimolar amount of 1-(((1- (4-amino-2-isopropoxyphenyl) piperidin-4-yl) (methyl) Amino) methyl) cyclopropane-1-ol instead of 1-((1- (4-amino-2-ethoxyphenyl) piperidin-4-yl) (methyl) amino) -2-methylpropane -2-ol.
  • Step 8) (2-((2-((4- (4-(((1-hydroxycyclopropyl) methyl) (methyl) amino) piperidin-1-yl) -3-isopropoxy Preparation of phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 4) (2-((2-((4- (4-(((1-hydroxycyclobutyl) methyl) (methyl) amino) piperidin-1-yl) -3-isopropoxy Phenyl) amino) -7-((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) di
  • step 6 For the preparation method, refer to step 6) in Example 17, wherein in an equimolar amount of 1-(((1- (4-amino-2-isopropoxyphenyl) piperidin-4-yl) (methyl) Amino) methyl) cyclobut-1-ol instead of 1-((1- (4-amino-2-ethoxyphenyl) piperidin-4-yl) (methyl) amino) -2-methylpropane -2-ol.
  • Step 5 (2-((2-((4- (4-(((1-hydroxycyclobutyl) methyl) (methyl) amino) piperidin-1-yl) -3-isopropoxy Preparation of phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 2) to Step 4) (2-((2-((4- (4- (cyclopropyl (methyl) amino) piperidin-1-yl) -3-isopropoxyphenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 2) to Step 4) Referring to Step 5) to Step 7) in the preparation method of Example 17, the starting material is equimolar equivalent of N-cyclopropyl-1- (2-isopropoxy-4- Nitrophenyl) -N-methylpiperidine-4-amine instead of 1-((1- (2-ethoxy-4-nitrophenyl) piperidin-4-yl) (methyl) amino) 2-methylpropan-2-ol.
  • Example 38 (2-((2-((4- (4- (4-((2-hydroxy-2-methylpropyl) (methyl) amino) piperidin-1-yl) -3- (trifluoro Preparation of methoxy) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 3 Dimethyl (2-((2-((4- (4- (methylamino) piperidin-1-yl) -3- (trifluoromethoxy) phenyl) amino) -7H- Preparation of pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) phosphine oxide
  • Step 4) (2-((2-((4- (4-((2-hydroxy-2-methylpropyl) (methyl) amino) piperidin-1-yl) -3- (trifluoromethyl Preparation of oxy) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Example 39 For the preparation method of Example 39, refer to the preparation steps 1) to 3) of Example 38, wherein in step 1), an equimolar equivalent of N, N-dimethylpiperidine-4-amine is used instead of tert-butylmethyl (piperidine Pyridin-4-yl) carbamate.
  • Step 3) to Step 5) (2-((2-((4- (4-((2-fluoroethyl) (methyl) amino) piperidin-1-yl) -3-isopropoxybenzene ) Amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 3) to Step 5) Referring to Step 5) to Step 7) in the preparation method of Example 17, the starting material is equimolar equivalent of N- (2-fluoroethyl) -1- (2-isopropoxy 4-nitrophenyl) -N-methylpiperidine-4-amine instead of 1-((1- (2-ethoxy-4-nitrophenyl) piperidin-4-yl) (methyl Group) amino) -2-methylpropan-2-ol.
  • Step 5) to Step 7) (2-((2-((3- (2-fluoroethoxy) -4- (4-((2-hydroxy-2-methylpropyl) (methyl) amino ) Piperidin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) amino) phenyl) dimethylphosphine
  • Step 5) to Step 7) Referring to Step 5) to Step 7) in the preparation method of Example 17, the starting material is an equimolar equivalent of 1-((1- (2- (2-fluoroethoxy)- 4-nitrophenyl) piperidin-4-yl) (methyl) amino) -2-methylprop-2-ol instead of 1-((1- (2-ethoxy-4-nitrophenyl ) Piperidin-4-yl) (methyl) amino) -2-methylpropan-2-ol.
  • EDTA (0.5M pH8.0) solution preparation accurately weigh 14.612g of EDTA powder, add ultrapure water and make up to 100mL (if insoluble, heat to 37 °C, adjust pH to 8.0 with NaOH solution)
  • 1 ⁇ Kinase Assay Buffer Add 25mL HEPES solution (1M), 190.175mg EGTA, 5mL MgCl 2 solution (1M), 1mL DTT, 50 ⁇ L Tween-20 to the reagent bottle, add ultrapure water to volume to 500mL (adjust pH) To 7.5).
  • 1 ⁇ Detection Buffer Take 1 mL of 10 ⁇ Detection Buffer and add 9 mL of water to mix.
  • EGFR T790M kinase solution Dilute the kinase stock solution to a concentration of 0.2nM with 1 ⁇ Kinase Assay Buffer, mix well, and store on ice.
  • substrate solution Dilute the substrate ULight TM -PolyGT stock solution to 400 nM with 1 ⁇ Kinase Assay Buffer and mix.
  • 4 ⁇ ATP solution Dilute the ATP stock solution to a concentration of 20 ⁇ M with 1 ⁇ Kinase Assay buffer, and mix well.
  • test method is as follows:
  • a 96-well plate a the compound with a concentration of 10 mM was diluted 3 times with DMSO solution to form 11 gradients, and the 12th gradient was a pure DMSO solution (as a positive control); a new 96-well plate b was taken, The above solution was diluted 25-fold with ultrapure water (DMSO concentration was 4%).
  • Add 2 ⁇ substrate / ATP mixed solution Use a row gun to take 5 ⁇ l of the above 2 ⁇ substrate / ATP mixed solution into the corresponding reaction wells of a 384-well plate.
  • Negative control set a negative control well in a 384-well plate, and add 2.5 ⁇ l 4 ⁇ substrate, 2.5 ⁇ l 4 ⁇ enzyme solution, 2.5 ⁇ l 1 ⁇ Kinase Assay Buffer and 2.5 ⁇ l ultrapure water containing 4% DMSO .
  • Inhibition rate (read value of positive control well-read value of experimental well) / (read value of positive control well-read value of negative control well) * 100%
  • test compound inhibition of EGFR T790M- L858R-C797S enzymatic reaction by calculating inhibition of EGFR T790M-L858R-C797S IC 50 to an enzymatic reaction Screen for compounds with biochemical inhibitory activity.
  • the specific method of the test is the same as the method of Experimental Example 1 above, only the following conditions need to be adjusted: 1).
  • the final concentration of the ATP solution in the reaction is 1 ⁇ M; 2).
  • the final concentration of the substrate ULight TM -PolyGT is 50 nM;
  • the final concentration of EGFR L858R-T790M-C797S kinase is 0.5 nM.
  • Table 2 lists the measurement results of the EGFR T790M kinase and EGFR L858R-T790M-C797S tyrosine kinase inhibitory activity of the compounds of the examples in the present application, where A represents an IC 50 of less than or equal to 5 nM, and B represents an IC 50 of greater than 5 nM but less than Or equal to 50nM, C means IC 50 is greater than 50nM, NT means no corresponding kinase was tested.
  • Table 4 lists the test results of the compounds of the embodiments of the present invention on Ba / F3 EGFR-L858R / T790M and Ba / F3 EGFR-L858R / T790M / C797S cells, where A represents an IC 50 of 50 nM or less, and B represents IC 50 is greater than 50 nM but less than or equal to 100 nM, C means IC 50 is greater than 100 nM but less than or equal to 200 nM, and D means IC 50 is greater than 200 nM.
  • test of small molecule compounds inhibiting ALK kinase activity is as follows:
  • Perkin Elmer Ultra TR-FRET technology measures the activity of ALK kinase substrate phosphorylation in the reaction.
  • LANCE Ultra enzyme activity analysis Eu-labeled specific anti-phospho antibodies were used to detect phosphorylation of Ulight-substrate. When the detection antibody binds to the phosphorylated substrate, Eu and Ulight are close to each other. After receiving laser radiation at a wavelength of 320 or 340 nm, the energy emitted by Eu is transferred to Ulight and a 665 nm optical signal is excited. The intensity of the light signal is positively correlated with the phosphorylation level of Ulight-substrate in the reaction, enabling the quantitative determination of kinase activity.
  • the enzyme detection experiment uses ALK kinase provided by Carna Biosciences, ATP provided by Promega, and Perkin Elmer's Ultra Ulight TM- Poly GT substrate, Eu-W1024-anti-phosphotyrosine detection antibody, LANCE TM Detection Buffer, white OptiPlate-384 well plate, sealing membrane and Envison multi-function plate reader were used for experiments and detection.
  • the preparation method of the kinase buffer solution, stop solution, and color development solution, and the method of compound dilution are as follows:
  • test solution Eu-W1024-anti-phosphotyrosine detection antibody was formulated at a concentration of 8 nM with 1 ⁇ Detection Buffer.
  • stop solution 0.8mL of EDTA (0.5M, pH8.0) aqueous solution, 1mL of 10 ⁇ Detection buffer and 8.2mL of ultrapure water were mixed.
  • ALK kinase, ATP, and Ultra Ulight TM- Poly GT substrate was prepared with kinase buffers at 4 ⁇ intermediate concentrations of 2.8nM, 4 ⁇ M, and 200nM, respectively.
  • the complete enzymatic reaction system includes the above 2.5 ⁇ L ALK kinase, 2.5 ⁇ L ATP, and 2.5 ⁇ L after dilution. 4 ⁇ intermediate concentration compounds and 2.5 ⁇ L Ultra Ulight TM- Poly GT substrate.
  • the reaction was protected from light at room temperature for 2 hours.
  • 5 ⁇ L of the stop solution was added to stop the enzymatic reaction, and then 5 ⁇ L of the color detection solution was added to the reaction for 1 hour.
  • Use Envison multi-function plate reader to call the corresponding program to read the plate.
  • Percent inhibition (read from positive control well-read from experimental well) / (read from positive control well-read from negative control well)] * 100%.
  • the drug concentration and the corresponding inhibition rate were input to GraphPad and Prism5 to calculate the corresponding IC50.
  • Table 5 lists the results of determination of ALK kinase inhibitory activity of some of the compounds of the examples in the present application, where A means that the IC50 is less than or equal to 5nM, B means that the IC50 is greater than 5nM but less than or equal to 50nM, and C means that the IC50 is greater than 50nM.

Abstract

本发明提供了一种含有芳胺基取代的吡咯并嘧啶类化合物、制备方法及其应用，具体涉及式(I)所示化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药，其制备方法及其在制备作为激酶抑制剂的药物中的应用。

Description

一种含有芳胺基取代的吡咯并嘧啶类化合物、制备方法及其应用 技术领域
本发明属于医药技术领域，涉及一种含有芳胺基取代的吡咯并嘧啶类化合物、制备方法及其应用。
背景技术
蛋白激酶是细胞生命活动重要的信号使者，可催化将ATP末端的γ-磷酸基团转移至底物氨基酸残基(丝氨酸、苏氨酸、酪氨酸)中的羟基受体上，从而激活目标蛋白(Johnson L.N.,and Lewis R.J.,Structural basis for control by phosphorylation,Cheminform,2001,101,2209.)。蛋白激酶参与了众多的生理过程，包括细胞增殖、存活、凋亡、代谢、转录以及分化等(Adams J.A.,Kinetic and catalytic mechanisms of protein kinases,Chemical reviews,2001,101,2271.)。在人体现有药物靶点中，蛋白激酶家族成员占比高达10％(Santos R.,Ursu O.,Gaulton A.,et al.,A comprehensive map of molecular drug targets,Nature Reviews Drug Discovery,2017,16,19.)。
表皮生长因子受体(ErbB)酪氨酸激酶可通过多种途径调节细胞增殖、迁移、分化、凋亡以及细胞移动。在多种形式的恶性肿瘤中，ErbB家族成员以及其部分配体通常过表达、扩增或突变，这使其成为重要的治疗靶标。该家族蛋白激酶包括：ErbB1/EGFR/HER1、ErbB2/HER2、ErbB3/HER3和ErbB4/HER4。其中EGFR是开发非小细胞肺癌的重要靶点(Dienstmann R.,et.al.,Personalizing Therapy with Targeted Agents in Non-Small Cell Lung Cancer,ONCOTARGET,2001,2(3),165.)。
Figure PCTCN2019088316-appb-000001
吉非替尼(Gefitinib)、埃罗替尼(Erlotinib)、埃克替尼(Icotinib)是第一代靶向EGFR的可逆型激酶抑制剂，用于治疗非小细胞癌。该类抑制剂同时对野生型和激活突变型EGFR具有抑制作用，且在临床上取得了较大的成功，但是受体患者服用一段时间后耐药性的出现，尤其是T790M突变导致的耐药性使疗效降低或失效。第二代EGFR抑制剂阿法替尼(Afatinib)为非可逆型抑制剂，其含有迈克尔受体，可与位于ATP结合口袋入口处的半胱氨酸残基(Cys797)发生共价键结合，该抑制剂针对T790M突变型EGFR激酶和野生型EGFR激酶均表现出极强的活性，且对于T790M突变型EGFR激酶的抑制活性高于野生型EGFR激酶，这使得该药物临床应用中治疗窗口较窄，使用效果并不理想(Camidge,D.R.,et.al.,Acquired resistance to TKIs in solid tumours:learning from lung cancer.Nature Reviews Clinical Oncology,2014,11,473.)。第三代的EGFR激酶抑制剂奥希替尼(Osimertinib)和奥莫替尼(Olmutinib)实现了对T790M突变型EGFR激酶相比野生型EGFR激酶的高选择性抑制，拉大了临床使用窗口，实现了对T790M突变病人的有效治疗。不幸的是，三代EGFR激酶抑制剂在临床上使用一段时间后也产生了耐药现象。其原因之一，是由于EGFR产生了C797S的二次突变。现有三代小分子EGFR抑制剂与靶标的作用机制是药物分子与EGFR的Cys797形成共价结合。但病人体内发生C797S的二次突变时，药物分子丧失了与Cys797的共价结合，导致药物的失效(Harun Patel.,et.al.,2017,Recent updates on third generation EGFR inhibitors and emergence of fourth generation EGFR inhibitors to combat C797S resistance,Eur J Med Chem.,2017,142,32)。基于此，开发具有对T790M突变型EGFR激酶良好的抑制活性，同时具有对C797S突变型EGFR激酶良好抑制活性的新型药物分子具有重要意义。
Brigatinib是一种靶向的ALK抑制剂，由Ariad医药公司研发并于2017年获美国FDA批准上市用于治疗ALK阳性的非小细胞肺癌。根据文献报道，Brigatinib对C797S突变型EGFR激酶具有一定的抑制作用。
发明内容
鉴于上述讨论的内容，本发明旨在提供一种含有芳胺基取代的吡咯并嘧啶类化合物、其异构体、水合物、溶剂化物、药学上可接受的盐或前药，及其药物组合物，并涉及它们在制备作为激酶抑制剂的药物中的应用。
本发明的一个方面提供了一种式(I)所示化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药，
Figure PCTCN2019088316-appb-000002
式(I)中，
X为CH或N；
Y为CH或N；
R 1为-NHR 5，R 5为未被取代或被取代的芳基、杂芳基、环烷基、环烷基桥环结构、并环结构，
R 5中取代的芳基、杂芳基、环烷基、环烷基桥环结构、并环结构中的取代基为-CF 3、-OCF 3、羟基、氰基、卤素、C 1-C 6烷基、C 3-C 5环烷基、C 1-C 6烷氧基、C 3-C 5环烷基氧基、-S(＝O) 2R 6、-C(＝O)R 6、-P(＝O)R 6R 7、-S(＝O) 2NR 6R 7，
R 6和R 7分别独立地为-H、C 1-C 6烷基、C 3-C 6环烷基，
所述并环结构选自芳环并5-6元杂芳环基、5-6元杂芳环并5-6元杂芳环基、芳环并5-6元环烷基、芳环并5-6元杂环基、5-6元杂芳环并5-6元环烷基或5-6元杂芳环并5-6元杂环基；
R 2为-H、-CF 3、-CH 2CF 3、C 1-C 6烷基、C 3-C 6环烷基、C 3-C 4环烷基取代的C 1-C 2烷基、含一个氧原子的4-6元杂环基、或-(CH 2) mR 8，其中m为1、2、3整数，
R 8为-OH、-CN、-C(O)NH 2、-S(＝O) 2CH 3、C 1-C 3烷氧基、C 1-C 3烷硫基；
R 3和R 4分别独立地为-H、C 1-C 6烷基、C 3-C 6环烷基、含一个氮原子或含一个氧原子的4-6元杂环基或-(CH 2) nR 9,其中n为1、2、3整数，
所述含一个氮原子或含一个氧原子的4-6元杂环为非取代或被C 1-C 3的烷基所取代，
R 9为-OH、-CN、-C(O)NH 2、-S(＝O) 2CH 3、-NR'R”、C 1-C 3烷氧基、C 1-C 3烷硫基，
R'、R”分别独立地为H或C 1-C 3的烷基，
或者，R 3、R 4与其相连的氮原子构成4-6元杂环或6-9元螺环，所述杂环含1-2个选自N、O或S的杂原子或含基团-C(＝O)-或-S(＝O) 2-作为环成员，
所述R 3、R 4与其相连的氮原子构成的4-6元杂环为未被取代或分别被1-2个选自卤素、氰基、羟基、氨基、C 1-C 3烷基、C 1-C 3烷氧基、卤代C 1-C 3烷基、氰基取代的C 1-C 3烷基、羟基取代的C 1-C 3烷基、C 1-C 3烷氧基取代的C 1-C 3烷基所取代，
所述R 3、R 4与其相连的氮原子构成的6-9元螺环为包含一个氮原子的单螺环。
在一些优选的实施方案中，
R 1为-NHR 5，R 5选自以下基团：
Figure PCTCN2019088316-appb-000003
Q为N或CH，
T为NH，O或S，
R 10选自-H、-OH、-F、-Cl、-Br、-CN、-CF 3、-OCF 3、甲基、乙基、丙基、异丙基、 环丙基、环丁基、甲氧基、乙氧基、丙氧基、异丙氧基、环丙基氧基、环丁基氧基、或以下基团：
Figure PCTCN2019088316-appb-000004
R 11为-H、甲基、乙基、丙基、异丙基，
R 12为-H、-F、-Cl、-Br、羟基、氰基、三氟甲基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基。
在本申请的一些优选实施方案中，X为CH；Y为CH。
在本申请的一些优选实施方案中，R 1为
Figure PCTCN2019088316-appb-000005
在一些优选的实施方案中，R 2选自-H、-CF 3、-CH 2CF 3、甲基、乙基、丙基、异丙基、1-甲基丙基、2-甲基丙基、环丙基、环丁基、环戊基、环己基、环丙基甲基、环丙基乙基、环丁基甲基、环氧丙烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基。
在另一些优选的实施方案中，R 2选自正丁基、正戊基、异戊基、新戊基、正己基、异己基。
优选地，R 3和R 4分别独立地选自-H、甲基、乙基、丙基、异丙基、环丙基、环丁基、环戊基、环氧丙烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基、甲氨基乙基、二甲氨基乙基、甲氨基丙基、二甲氨基丙基，或R 3、R 4与其相连的氮原子构成4-6元杂环或6-9元螺环，所述4-6元杂环为取代或非取代的杂环，
所述R 3、R 4与其相连的氮原子构成的取代或非取代的4-6元杂环选自以下环结构：
Figure PCTCN2019088316-appb-000006
R 13选自–H、甲氨基、乙氨基、二甲氨基，
R 14选自-H、甲基、乙基、丙基、异丙基、甲酰基、乙酰基或甲磺酰基，
R 15和R 16分别独立地选自-H、-F、-CF 3、羟基、氨基、氰基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基、异丙氧基、氰基甲基、氰基乙基、甲氧基甲基、甲氧基 乙基、甲氧基丙基、羟甲基、羟乙基、羟丙基，
所述R 3、R 4与其相连的氮原子构成的6-9元螺环选自以下螺环结构：
Figure PCTCN2019088316-appb-000007
优选地，还可以包括如下方案，R 3选自-H、甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基、环丙基、环丁基、环戊基、环氧丙烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基、甲氨基乙基、二甲氨基乙基、甲氨基丙基、二甲氨基丙基，
R 4选自正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基。
本发明的另一方面提供一种式(I)所示化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药，
Figure PCTCN2019088316-appb-000008
式(I)中，
X为CH或N；Y为CH或N；优选地，X为CH；Y为CH；
R 1为-NHR 5，R 5选自以下基团：
Figure PCTCN2019088316-appb-000009
Q为N或CH，
T为NH，O或S，
R 10选自-H、-OH、-F、-Cl、-Br、-CN、-CF 3、-OCF 3、甲基、乙基、丙基、异丙基、环丙基、环丁基、甲氧基、乙氧基、丙氧基、异丙氧基、环丙基氧基、环丁基氧基、或以下基团：
Figure PCTCN2019088316-appb-000010
R 11为-H、甲基、乙基、丙基、异丙基，
R 12为-H、-F、-Cl、-Br、羟基、氰基、三氟甲基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基；
优选地，R 1为
Figure PCTCN2019088316-appb-000011
R 2选自-H、-CF 3、-CH 2CF 3、甲基、乙基、丙基、异丙基、1-甲基丙基、2-甲基丙基、环丙基、环丁基、环戊基、环己基、环丙基甲基、环丙基乙基、环丁基甲基、氧杂环丁烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基；
R 3选自-H、甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基、环丙基、环丁基、环戊基、氧杂环丁烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基；
R 4选自-(CH 2) nR 17,其中n为1、2、3整数，
R 17为
Figure PCTCN2019088316-appb-000012
或者C 3-C 6环烷基，R 18为-OH、-CN、-C(O)NH 2、
-S(＝O) 2CH 3、C 1-C 3烷氧基、C 1-C 3烷硫基。
本发明的另一方面提供一种式(I)所示化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药，
Figure PCTCN2019088316-appb-000013
式(I)中，
X为CH或N；
Y为CH或N；
R 1为-NHR 5，R 5选自以下基团：
Figure PCTCN2019088316-appb-000014
Q为N或CH，
T为NH、O或S，
R 10选自-H、-OH、-F、-Cl、-Br、-CN、-CF 3、-OCF 3、甲基、乙基、丙基、异丙基、环丙基、环丁基、甲氧基、乙氧基、丙氧基、异丙氧基、环丙基氧基、环丁基氧基、或以下基团：
Figure PCTCN2019088316-appb-000015
R 11为-H、甲基、乙基、丙基、异丙基，
R 12为-H、-F、-Cl、-Br、羟基、氰基、三氟甲基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基；
R 2为-H、C 1-C 6烷基、C 3-C 6环烷基、卤代C 1-C 6烷基；
R 3和R 4分别独立地为-H、C 3-C 8环烷基、或由1至3个选自C 1-C 6烷氧基、羟基、卤素、C 3-C 6环烷基的取代基所取代或者非取代的C 1-C 9烷基，
或者，R 3、R 4与其相连的氮原子构成4-8元杂环，所述杂环还含1-2个选自N、O或S的杂原子或含基团-C(＝O)-或-S(＝O) 2-作为环成员，
所述R 3、R 4与其相连的氮原子构成的4-8元杂环为未被取代或被1-2个选自卤素、氰基、羟基、氨基、C 1-C 3烷基、C 1-C 3烷氧基、卤代C 1-C 3烷基、氰基取代的C 1-C 3烷基、羟基取代的C 1-C 3烷基、C 1-C 3烷氧基取代的C 1-C 3烷基所取代。
在一些优选的实施方案中，X为CH；Y为CH；
R 1为
Figure PCTCN2019088316-appb-000016
R 2为-H、甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、异戊基、戊基、新戊基、己基、异己基、环丙基、环丁基、环戊基、环己基、三氟甲基、氟乙基；
R 3和R 4分别独立地选自-H、甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基、环丙基、环丁基、环戊基、环己基、氧杂环丁烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基、甲硫基乙基、甲硫基丙基、甲氧基甲基、甲氧基乙基、甲氧基丙基、乙氧基甲基、乙氧基乙基、乙氧基丙基、异 丙氧基甲基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟甲基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基、甲氨基乙基、二甲氨基乙基、甲氨基丙基、二甲氨基丙基、环丙基甲基、环丙基乙基、三氟甲基、氟乙基、2-羟基-2-甲基丙基、3-羟基-3-甲基丁基、
Figure PCTCN2019088316-appb-000017
或R 3、R 4与其相连的氮原子构成4-8元杂环，所述4-8元杂环为取代或非取代的杂环，
所述R 3、R 4与其相连的氮原子构成的取代或非取代的4-8元杂环选自以下环结构：
Figure PCTCN2019088316-appb-000018
R 13选自–H、甲氨基、乙氨基、二甲氨基，
R 14选自-H、甲基、乙基、丙基、异丙基、甲酰基、乙酰基或甲磺酰基，
R 15和R 16分别独立地选自-H、-F、-CF 3、羟基、氨基、氰基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基、异丙氧基、氰基甲基、氰基乙基、甲氧基甲基、甲氧基乙基、甲氧基丙基、羟甲基、羟乙基、羟丙基。
本申请包括但不限于如下典型化合物：
Figure PCTCN2019088316-appb-000019
Figure PCTCN2019088316-appb-000020
根据本发明的又一方面，提供一种制备所述化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药的方法，包括以下步骤，
Figure PCTCN2019088316-appb-000021
PG选自：三甲硅基乙氧基甲基、叔丁氧羰基、苄氧羰基、对甲苯磺酰基、苯磺酰基、乙酰基、三氟乙酰基、芴甲氧羰基、苄基；优选为三甲硅基乙氧基甲基(SEM基)，
其中，R 1、R 2、R 3、R 4如前文所述。
进一步地，本申请提供一种治疗与酪氨酸激酶EGFR，HER2，或ALK突变或过表达相关疾病的药用组合物，其由所述的式(I)的化合物或其药学上可接受的盐或其水合物或其溶剂化物或其前药与药学上可接受的载体或赋形剂组成。
根据本申请的又一方面，提供一种药用组合物:其中包含如所述的式(I)的化合物或其药学上可接受的盐、水合物、溶剂化物、或前药作为活性成分，一个或多个其它的治疗剂，以及一种或多种药学上可接受的载体或赋形剂。
本申请还涉及所述的式(I)的化合物、其异构体、水合物、溶剂化物、药学上可接受的盐或前药在制备治疗与酪氨酸激酶EGFR，HER2，或ALK突变或过表达相关的癌症及自身免疫疾病的药物中的应用，其中所述癌症及自身免疫疾病包括眼底疾病、干眼症、银屑病、白癜风、皮炎、斑秃、类风湿性关节炎、结肠炎、多重硬化、***性红斑狼疮、克罗恩病、动脉粥样化、肺纤维化、肝纤维化、骨髓纤维化、非小细胞肺癌、小细胞肺癌、乳腺癌、胰腺癌、神经胶质瘤、胶质母细胞瘤、卵巢癌、子***、结肠直肠癌、黑色素瘤、子宫内膜癌、***癌、膀胱癌、白血病、胃癌、肝癌、胃肠间质瘤、甲状腺癌、慢性粒细胞白血病、急性髓细胞性白血病、非霍奇金淋巴瘤、鼻咽癌、食道癌、脑瘤、B细胞和T细胞淋巴瘤、淋巴瘤、多发性骨髓瘤、胆道癌肉瘤、胆管癌中的任一种。
本申请还涉及一种治疗EGFR、HER2或ALK等激酶介导的疾病或病症的方法，其包括对有需要的患者(人或其他哺乳动物，尤其是人)给药治疗有效量的式(I)化合物或其盐，所述EGFR、HER2或ALK等激酶介导的疾病或病症包括前述提及的那些。
具体实施方式
除非另有说明，在本申请(包括说明书和权利要求书)中使用的以下术语具有下面给出的定义。在本申请中，除非另外说明，使用“或”或“和”意味着“和/或”。此外，术语“包括”以及其它形式的使用，例如“包含”、“含有”和“具有”，不是限制性的。本文使用的章节标题仅仅是为了组织的目的，而不应解释为对所述的主题的限制。
在这里所指的术语“取代”，包括复杂取代基(比如，芳基(如苯基)，杂烷基，杂芳基)，比较合适的是1至5个取代基，较好的是1到3个，最好是1到2个，可从取代基列表上自由选择。
除非有特殊说明，烷基表示具有指定数目碳原子的饱和直链、支链烃基，术语C 1-C 10烷基表示含有1至10个碳原子的烷基部分，同理C 1-C 3烷基表示含有1至3个碳原子的烷 基部分，比如，C 1-C 6烷基包括甲基、乙基、丙基、异丙基、n-丁基、异丁基、仲-丁基、叔-丁基、n-戊基、3-(2-甲基)丁基、2-戊基、2-甲基丁基、新戊基、n-己基、2-己基和2-甲基戊基等。
当取代基术语例如“烷基”与其它取代基术语组合使用时，例如在术语“C 1-C 3烷氧基C 1-C 6烷硫基”或“羟基取代C 1-C 10烷基”中，该连接取代基术语(例如烷基或烷硫基)旨在包含二价的部分，其中连接点通过所述连接取代基。“C 1-C 3烷氧基C 1-C 6烷硫基”的实例包括但不限于甲氧基甲硫基、甲氧基乙硫基和乙氧基丙硫基等。“羟基取代C 1-C 10烷基”的实例包括但不限于羟基甲基、羟基乙基和羟基异丙基等。
烷氧基由先前描述的直链或支链烷基与-O-形成的烷基-O-基团，例如，甲氧基、乙氧基等等。类似的，烷硫基由先前描述的直链或支链烷基与-S-形成的烷基-S-基团，例如，甲硫基，乙硫基等等。
烯基和炔基包括直链、支链烯基或炔基，术语C 2-C 6烯基或者C 2-C 6炔基表示具有至少一个烯基或炔基的直链或支链烃基。
“环烷基”表示含有指定数目碳原子的非芳香的、饱和的、环状的烃基。例如，术语“(C3-C6)环烷基”指的是具有3-6个环碳原子的非芳香的环状烃环。示例性的“(C3-C6)环烷基”包括环丙基、环丁基、环戊基和环己基。
在这里使用的术语“芳基”，除非有特别说明，指的是未被取代的或已被取代的芳香基，例如苯基，萘基，蒽基。术语“芳酰基”指-C(O)-芳基。
在这里使用的术语“杂环基”，除非有特殊说明，代表未被取代的或已被取代的稳定的3至8元单环饱和环体系，它们由碳原子以及从N，O，S中选的1至3个杂原子组成，其中N，S杂原子可以被随意氧化，N杂原子还可以被随意季铵化。杂环可以和任何杂原子或碳原子结合，从而组成一个稳定的结构。这类杂环的例子包括(但并不局限于)氮杂环己烷基，吡咯烷基，哌啶基，哌嗪基，氧化哌嗪基，氧化哌啶基，四氢呋喃基，二氧戊环基，四氢咪唑基，四氢噻唑基，四氢噁唑基，四氢吡喃基，***啉基，硫代***啉基，噻***啉亚砜，噻***啉砜以及噁二唑基。
在这里使用的术语“杂芳基”，除非有特别说明，代表未被取代或已被取代的稳定的5或6元单环芳香环体系，也可以代表未被取代或已被取代的9或10元苯稠苯杂芳环体系或二环杂芳环体系，它们由碳原子和由1至4个从N，O，S中选择的杂原子组成，其中N，S杂原子可以被随意氧化，N杂原子还可以被随意季铵化。杂芳基可以和任何杂原子或碳原子黏附起来，从而组成一个稳定的结构。杂芳基的例子包括(但并不局限于)噻蒽基，呋喃基，咪唑基，异噁唑基，噁唑基，吡唑基，吡咯基，噻唑基，噻二唑基，***基，吡啶基，哒嗪基，吲哚基，氮杂吲哚基，吲唑基，苯并咪唑基，苯并呋喃基，苯并噻吩基，苯并异噁唑基，苯并噁唑基，苯并吡唑基，苯并噻唑基，苯并噻二唑基，苯并***基，腺嘌呤基，喹啉基或异喹啉基。
术语“羰基”指的是C(O)基。
无论何时术语“烷基”或“芳基”或任何它们的前缀词根出现在一个取代物的名称中(例如，芳烷基，二烷基氨)，它将被认为包含了以上为“烷基”和“芳基”而给出的那些限制。碳原子的指定数量(比如，C1-C6)将独立的表示在一个烷基部分或在一个更大的取代基中的烷基部分(其中烷基作为其前缀词根)中的碳原子的数量。
很清楚，式I的化合物、异构体、晶型或前药及其可药用盐可以存在溶剂化形式和非溶剂化形式。例如溶剂化形式可以是水溶形式。本发明包括所有这些溶剂化的和未溶剂化的形式。
本发明的化合物可能有不对称的碳原子，根据它们的理化差异，通过已知技术上已成熟的方法，比如，通过色谱或分步结晶法，这种非对映异构的混合物可以被分离成单一的非对映异构体。对映异构体的分离可通过先用适当有旋光活性的化合物进行反应，把对映异构的混合物转化成非对映异构的混合物，分离非对映异构体，再把单一非对映异构体转化(水解)成相应的纯的对映异构体。所有这样的异构体，包括非对映异构体混合物和纯对映体被认为是该发明的一部分。
作为活性成分的本发明的化合物，以及制备该化合物的方法，都是本发明的内容。而且，一些化合物的晶型形式可以作为多晶体存在，这种形式也可以被包括在目前的发明里。另外，一些化合物可以和水(即水合物)或普通的有机溶剂一起形成溶剂化物，这种溶剂化物也被包括在此项发明的范畴内。
本发明的化合物可以以游离的形式用于治疗，或者在适当情况下以药学上可接受的盐或其它衍生物的形式用于治疗。如本文所用，术语“药学上可接受的盐”是指本发明的化合物的有机盐及无机盐，此盐适用于人类和低等动物，无过度毒性、刺激性、过敏反应等，具有合理的利益/风险比。胺，羧酸，膦酸盐，和其它类型的化合物的药学上可接受的盐在所属领域中是众所周知的。该盐可以由本发明的化合物与合适的游离碱或酸反应而成。包括但不限于，与无机酸如盐酸、氢溴酸、磷酸、硫酸、高氯酸或与有机酸如乙酸、草酸、马来酸、酒石酸、柠檬酸、琥珀酸、丙二酸形成的盐，或通过使用本领域熟知的方法，例如离子交换法，来得到这些盐。其他药学上可接受的盐包括己二酸盐、藻酸盐、抗坏血酸盐、天冬氨酸盐、苯磺酸盐、苯甲酸盐、硫酸氢盐、硼酸盐、丁酸盐、樟脑酸盐、樟脑磺酸盐、柠檬酸盐、二葡糖酸盐、十二烷基硫酸盐、乙磺酸盐、甲酸盐、富马酸盐、葡庚糖酸盐、甘油磷酸盐、葡萄糖酸盐、半硫酸盐、己酸盐、氢碘酸盐、2-羟基乙磺酸盐、乳糖酸盐、乳酸盐、月桂酸盐、月桂基硫酸盐、苹果酸盐、马来酸盐、甲烷磺酸盐、2-萘磺酸盐、烟酸盐、硝酸盐、油酸盐、棕榈酸盐、双羟萘酸盐、果胶酸盐、过硫酸盐、过3-苯基丙酸盐、磷酸盐、苦味酸盐、丙酸盐、硬脂酸盐、硫酸盐、硫氰酸盐、对甲苯磺酸盐、十一烷酸盐等。代表性的碱或碱土金属盐包括钠、锂、钾、钙、镁等。其他药学上可接受的盐包括适当的无毒的铵、季铵，和使用诸如卤离子、氢氧根、羧酸根、硫酸根、磷酸根、硝酸根，低级烷基磺酸盐和芳基磺酸盐形成的胺基阳离子。
另外，本文所用术语“前药”是指一个化合物在体内可以转化为本发明式(I)所示的化合物。此转化受前体药物在血液中水解或在血液或组织中经酶转化为母体化合物的影响。
本发明的药物组合物包含本文所述结构式(I)化合物或其药学上可接受的盐、激酶抑制剂(小分子，多肽，抗体等)、免疫抑制剂、抗癌药、抗病毒剂、抗炎剂、抗真菌剂、抗生素或抗血管过度增生化合物的另外的活性剂；以及任何药学上可接受的载体、佐剂或赋形剂。
本发明的化合物可以作为单独使用，也可以与一种或多种其它本发明的化合物或与一种或多种其它药剂联合使用。当联合给药时，治疗剂可以配制成同时给药或顺序地在不同的时间给药，或者所述治疗剂可以作为单一组合物给药。所谓“组合疗法”，指的是使用本发明的化合物与另一种药剂一起使用，给药方式为每种药剂同时共同给药或每种药剂顺序给药，无论哪种情况，目的都是要达到药物的最佳效果。共同给药包括同时递送剂型，以及每种化合物分别的单独剂型。因此，本发明的化合物的给药可以与已知的本领域的其他疗法同时使用，例如，在癌症治疗中使用放射治疗或细胞生长抑制剂、细胞毒性剂、其它抗癌剂等附加疗法来改善癌症状。本发明并不限于给药的顺序；本发明的化合物可以先前施用，同时施用，或在其他抗癌剂或细胞毒性剂之后施用。
为了制备这一发明的药学成分，作为其活性成分的分子式(I)的一种或多种化合物或盐类可紧密的与药学载体混合在一起，这是根据传统的制药配料技术而进行的，其中的载体可根据按不同的给药方式(例如，口服或肠外给药)设计好的制备形式而采用多种多样的形式。适当的药学上可接受的载体在技术上是众所周知的。对一些这类药学可接受的载体的描述可以在《药学赋形剂手册》里找到，该书由美国药学会和英国药学社联合出版。
本发明药物组合物可以有以下形式，比如说，适合口服给药，例如药片，胶囊，药丸，药粉，持续释放的形式，溶液或悬浮液；用于胃肠外注射如透明液，悬浮液，乳状液；或者用于局部用药如膏，霜；亦或作为栓剂用于直肠给药。药学成分也可以单位剂量的形式适合用于精确剂量的一次性给药。该药学成分将包括一种传统的药学载体或赋形剂以及根据目前的发明制成的作为活性成分的化合物，另外，也可以包括其他的医学或药学制剂，载体，辅助剂，等等。
治疗性化合物也可给于哺乳动物而非人类。给一个哺乳动物所用的药物剂量将取决于该动物的种类以及它的疾病状况或其所处的失调状态。治疗性化合物可以以胶囊，大丸药，药片药水的形式喂给动物。也可以通过注射或灌输的方式让治疗性化合物进入动物体内。我们根据符合兽医实践标准的传统的方式制备好这些药物形式。作为一种可选择的方式，药学合成药可以同动物饲料混合在一起喂给动物，因此，浓缩的饲料添加剂或预拌和料可以备以混合普通的动物饲料。
本发明的又一目的是在于提供一种用于治疗有需要的受试者中癌症的方法，其包括给受试者施用含本发明的化合物的组合物的治疗有效量的一种方法。
本发明还包括本发明的化合物或其药学上可接受的衍生物的使用，制备治疗与酪氨酸激酶EGFR、HER2、ALK相关的癌症及自身免疫疾病的药物中的应用。所述的癌症(包括非实体瘤、实体瘤、原发性或转移性癌症，如本文别处所指出和包括癌症具有抗性或难治的一种或多种其它治疗)以及其它疾病(包括但不限于眼底疾病、银屑病、动脉粥样化、肺纤维化、肝纤维化、骨髓纤维化等)的药剂。所述癌症包括但不限于：非小细胞肺癌、小细胞 肺癌、乳腺癌、胰腺癌、神经胶质瘤、胶质母细胞瘤、卵巢癌、子***、结肠直肠癌、黑色素瘤、子宫内膜癌、***癌、膀胱癌、白血病、胃癌、肝癌、胃肠间质瘤、甲状腺癌、慢性粒细胞白血病、急性髓细胞性白血病、非霍奇金淋巴瘤、鼻咽癌、食道癌、脑瘤、B细胞和T细胞淋巴瘤、淋巴瘤、多发性骨髓瘤、胆道癌肉瘤、胆管癌中的任一种。
本发明还提供了制备相应化合物的方法，可以使用多种合成方法制备本文所述的化合物，包括下述的方法，本发明的化合物或者其药学上可接受的盐，异构体或水合物可以使用下述方法与有机化学合成领域已知的合成方法，或通过本领域技术人员理解对这些方法的变化方法合成，优选方法包括但不限于下述方法。
一种制备本发明化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药的方法，包括以下步骤:
Figure PCTCN2019088316-appb-000022
其中，R 1、R 2、R 3、R 4如前文所定义。
反应条件：
步骤1)中的式(IV)与式(III)所示化合物充分接触得到式(II)所示化合物；
优选地，该反应可在有机溶剂中进行，所述的有机溶剂包含但不限于：N,N-二甲基甲酰胺(DMF)、N,N-二甲基乙酰胺(DMA)、N-甲基-2-吡咯烷酮(NMP)、乙二醇二甲醚、异丙醇、正丁醇、2-丁醇(仲丁醇)、叔丁醇的一种或两种以上的组合；
优选地，该反应可在碱的存在下进行，所述的碱包含但不限于：碳酸钾、碳酸钠、醋酸钠、三乙胺、二异丙基乙基胺、三乙烯二胺、吡啶、4-二甲氨基吡啶、1,8-二氮杂二环十一碳-7-烯或N-甲基吗啉的一种或两种以上的组合；
优选地，该反应可在酸存在下进行，所述的酸包含但不限于：三氟乙酸、对甲苯磺酸；
优选地，该反应可在钯金属催化偶联反应条件下进行，所述的钯金属催化偶联反应条件为常见的Buchward-Hartwig反应所用的钯配体、溶剂及碱；
步骤2)中式(II)所示化合物在四丁基氟化铵存在的条件下得到式(I)所示化合物。
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合具体实施例，对本发 明进行进一步详细说明。应当理解，此处所描述的具体实施例仅用以解释本发明，并不用于限定本发明。实施例中未注明具体技术或条件的，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。本文所使用的术语“和/或”包括一个或多个相关的所列项目的任意的和所有的组合。下面提供的实施例可以更好的说明本发明，除非特别说明，所有的温度为摄氏度。
本申请中所使用的部分中间体的合成，参见中国专利申请号WO2016000581A。
实施例1.(2-((2-((4-(4-(二甲氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000023
步骤1)(2-((2-((4-(4-(二甲氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000024
将1-(4-氨基-2-甲氧基苯基)-N,N-二甲基哌啶-4-胺249毫克(1mmol)、(2-((2-氯-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧磷450毫克(1mmol)、对甲苯磺酸17毫克(0.1mmol)置于反应瓶中，加入10毫升仲丁醇，加热搅拌至反应完毕，旋蒸浓缩后柱层析得到产品330毫克，产率50％。MS:664[M+H] +.
步骤2)(2-((2-((4-(4-(二甲氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000025
将(2-((2-((4-(4-(二甲氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦330毫克(0.5mmol)置于反应瓶中，加入四氢呋喃溶解，随后加入1摩尔每升的四丁基氟化铵四氢呋喃溶液2毫升，搅拌至反应完毕，蒸去溶剂后柱层析得到产品265毫克，产率50％。 1H NMR(400MHz,DMSO-d6)δ11.62 (s,1H),11.24(s,1H),9.15(d,J＝5.3Hz,1H),8.76(s,1H),7.63–7.53(m,1H),7.48(s,2H),7.35(d,J＝8.7Hz,1H),7.06(t,J＝7.2Hz,1H),6.96–6.90(m,1H),6.81(d,J＝8.5Hz,1H),6.37(d,J＝2.6Hz,1H),3.78(s,3H),3.14–2.82(m,4H),2.22(s,6H),2.18–2.13(m,1H),1.83(d,J＝13.5Hz,6H),1.81–1.77(m,2H),1.58–1.47(m,2H)；MS:534[M+H] +.
实施例2.(2-((2-((4-(4-(二甲氨基)哌啶-1-基)-3-乙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000026
参考实施例1的制备，起始原料中以等摩尔当量的1-(4-氨基-2-乙氧基苯基)-N,N-二甲基哌啶-4-胺替代1-(4-氨基-2-甲氧基苯基)-N,N-二甲基哌啶-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.61(s,1H),11.22(s,1H),9.24–9.06(m,1H),8.73(s,1H),7.63–7.52(m,1H),7.52–7.42(m,2H),7.33(d,J＝8.5Hz,1H),7.13–7.00(m,1H),6.98–6.89(m,1H),6.79(d,J＝8.6Hz,1H),6.36(d,J＝2.9Hz,1H),4.12–3.92(m,2H),3.37(d,J＝11.3Hz,2H),3.20–2.79(m,2H),2.22(s,6H),2.19–2.12(m,1H),1.90–1.76(m,8H),1.60–1.45(m,2H),1.38–1.35(m,3H)；MS:548[M+H] +.
实施例3.(2-((2-((4-(4-(二甲氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000027
参考实施例1的制备，起始原料中以等摩尔当量的1-(4-氨基-2-异丙氧基苯基)-N,N-二甲基哌啶-4-胺替代1-(4-氨基-2-甲氧基苯基)-N,N-二甲基哌啶-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.60(s,1H),11.21(s,1H),9.14(s,1H),8.72(s,1H),7.63–7.47(m,2H),7.38(d,J＝12.5Hz,2H),7.07(t,J＝7.3Hz,1H),6.96–6.90(m,1H),6.80(d,J＝8.3Hz,1H),6.38–6.33(m,1H),4.68–4.48(m,1H),3.43–3.35(m,2H),3.02–2.93(m,2H),2.22(s,6H),2.03–1.94(m,1H),1.87–1.78(m,8H),1.59–1.43(m,2H),1.29(d,J＝6.0Hz,6H)；MS:562[M+H] +.
实施例4.(2-((2-((3-乙氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000028
参考实施例1的制备，起始原料中以等摩尔当量的3-乙氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯胺替代1-(4-氨基-2-甲氧基苯基)-N,N-二甲基哌啶-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.60(s,1H),11.22(s,1H),9.21–9.04(m,1H),8.72(s,1H),7.63–7.52(m,1H),7.52–7.41(m,2H),7.39–7.27(m,1H),7.06(t,J＝7.5Hz,1H),7.00–6.88(m,1H),6.79(d,J＝8.6Hz,1H),6.36(d,J＝2.9Hz,1H),4.08–3.93(m,2H),3.42–3.34(m,2H),3.22–2.74(m,2H),2.49–2.43(m,4H),2.43–2.28(m,4H),2.28–2.21(m,1H),2.17(s,3H),1.90–1.73(m,8H),1.63–1.46(m,2H),1.36(t,J＝6.8Hz,3H)；MS:603[M+H] +.
实施例5.(2-((2-((3-异丙氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000029
参考实施例1的制备，起始原料中以等摩尔当量的3-异丙氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯胺替代1-(4-氨基-2-甲氧基苯基)-N,N-二甲基哌啶-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.60(s,1H),11.21(s,1H),9.21–9.04(m,1H),8.71(s,1H),7.63–7.43(m,2H),7.38(d,J＝9.2Hz,2H),7.06(t,J＝7.2Hz,1H),6.99–6.89(m,1H),6.79(d,J＝8.4Hz,1H),6.41–6.28(m,1H),4.60–4.50(m,1H),3.38(d,J＝11.0Hz,2H),3.18–2.85(m,2H),2.48–2.39(m,4H),2.38–2.24(m,4H),2.24–2.18(m,1H),2.15(s,3H),1.88–1.69(m,8H),1.60–1.46(m,2H),1.29(d,J＝6.0Hz,6H).MS:617[M+H] +.
实施例6.(2-((2-((4-(4-(二甲基氨基)哌啶-1-基)-3-异丁氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000030
步骤1)1-氟-2-异丁氧基-4-硝基苯的制备
Figure PCTCN2019088316-appb-000031
将1-碘代-2-甲基丙烷184毫克(1mmol)、2-氟-5-硝基苯酚157毫克(1mmol)置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，搅拌至反应完毕，加水、抽滤得产品181毫克，产率85％。MS:214[M+H] +.
步骤2)1-(2-异丁氧基-4-硝基苯基)-N,N-二甲基哌嗪-4-胺的制备
Figure PCTCN2019088316-appb-000032
将N,N-二甲基哌嗪-4-胺108毫克(0.85mmol)、1-氟-2-异丁氧基-4-硝基苯181毫克(0.85mmol)置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，搅拌至反应完毕，加水、抽滤得产品245毫克，产率90％。MS:322[M+H] +.
步骤3)1-(4-氨基-2-异丁氧基苯基)-N,N-二甲基哌啶-4-胺的制备
Figure PCTCN2019088316-appb-000033
将1-(2-异丁氧基-4-硝基苯基)-N,N-二甲基哌嗪-4-胺245毫克(0.77mmol)置于反应瓶中，加入5毫升甲醇、铁粉431毫克(7.7mmol)和氯化铵407毫克(7.7mmol)加热至反应完毕，抽滤、加入碳酸钾水溶液调至碱性，乙酸乙酯萃取、浓缩得产品157毫克，产率70％。MS:292[M+H] +.
步骤4)和步骤5)(2-((2-((4-(4-(二甲基氨基)哌啶-1-基)-3-异丁氧基苯基)氨基)-7H吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000034
参考实施例1的步骤1)和步骤2)制备，起始原料中以等摩尔当量的1-(4-氨基-2-异丁氧基苯基)-N,N-二甲基哌啶-4-胺替代1-(4-氨基-2-甲氧基苯基)-N,N-二甲基哌啶-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.56(s,1H),11.16(t,J＝2.2Hz,1H),9.23–8.97(m,1H),8.67(s,1H),7.55–7.46(m,1H),7.44–7.37(m,1H),7.30(d,J＝7.9Hz,2H),7.02–6.95(m,1H),6.89–6.82(m,1H),6.71(d,J＝8.4Hz,1H),6.34–6.25(m,1H),3.64(d,J＝6.2Hz,2H),3.35–3.30(m,2H),2.53–2.45(m,2H),2.15(s,6H),2.12–2.04(m,1H),2.04–1.92(m,1H),1.86–1.68(m,8H),1.55–1.36(m,2H),0.96(d,J＝6.7Hz,6H)；MS:576[M+H] +.
实施例7.(2-((2-((3-环丁氧基-4-(4-(二甲基氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000035
实施例7的制备参考实施例6的步骤1)至步骤5)，步骤1)中的起始原料中以等摩尔当量的碘代环丁烷替代1-碘代-2-甲基丙烷。 1H NMR(400MHz,DMSO-d6)δ11.54(s,1H),11.13(t,J＝2.2Hz,1H),9.26–8.96(m,1H),8.67(s,1H),7.60–7.38(m,2H),7.33–7.27(m,1H),7.20(d,J＝2.4Hz,1H),7.04–6.96(m,1H),6.89–6.82(m,1H),6.71(d,J＝8.7Hz,1H),6.33–6.24(m,1H),4.61–4.47(m,1H),3.34–3.26(m,4H),2.42–2.36(m,4H),2.16(s,6H),2.06–1.91(m,2H),1.76(d,J＝13.5Hz,8H),1.66–1.53(m,1H),1.52–1.39(m,2H)；MS:574[M+H] +.
实施例8.(2-((2-((3-甲氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000036
实施例8的制备参考实施例6的制备路线的步骤1)至步骤5)，其中步骤1)中起始物料以等摩尔当量的碘甲烷替代1-碘代-2-甲基丙烷，步骤2)中以等摩尔当量的1-甲基-4-(哌啶-4-基)哌嗪替代N,N-二甲基哌嗪-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.56(s,1H),11.18(t,J＝2.0Hz,1H),9.33–8.99(m,1H),8.71(s,1H),7.56–7.46(m,1H),7.42(t,J＝4.6Hz,2H),7.32–7.24(m,1H),7.03–6.94(m,1H),6.90–6.81(m,1H),6.73(d,J＝8.6Hz,1H),6.33– 6.20(m,1H),3.81–3.59(m,6H),3.25–3.17(m,2H),2.59–2.45(m,3H),2.42–2.20(m,4H),2.20–2.13(m,1H),2.09(s,3H),1.76(d,J＝13.5Hz,6H),1.74–1.66(m,2H),1.55–1.40(m,2H).MS:589[M+H] +.
实施例9.(2-((2-((3-异丁氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000037
实施例9的制备参考实施例6的制备路线的步骤1)至步骤5)，其中步骤2)中以等摩尔当量的1-甲基-4-(哌啶-4-基)哌嗪替代N,N-二甲基哌嗪-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.61(s,1H),11.21(s,1H),9.18–9.00(m,1H),8.71(s,1H),7.62–7.51(m,1H),7.47(t,J＝8.0Hz,1H),7.36(d,J＝8.3Hz,2H),7.05(t,J＝7.1Hz,1H),6.92(t,J＝2.8Hz,1H),6.77(d,J＝8.4Hz,1H),6.39–6.28(m,1H),3.70(d,J＝6.2Hz,2H),3.40–3.35(m,4H),2.57–2.52(m,4H),2.39–2.26(m,4H),2.25–2.19(m,1H),2.15(s,3H),2.08–2.01(m,1H),1.89–1.77(m,8H),1.60–1.46(m,2H),1.03(d,J＝6.7Hz,6H)；MS:631[M+H] +.
实施例10.(2-((2-((3-环丁氧基-4-(4-(4-甲基哌嗪-1-基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000038
实施例10的制备参考实施例7的制备路线的步骤1)至步骤5)，其中步骤2)中以等摩尔当量的1-甲基-4-(哌啶-4-基)哌嗪替代N,N-二甲基哌嗪-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.59(s,1H),11.18(s,1H),9.15–9.07(m,1H),8.72(s,1H),7.60–7.46(m,2H),7.39–7.33(m,1H),7.26(d,J＝2.3Hz,1H),7.06(t,J＝7.5Hz,1H),6.92(t,J＝2.8Hz,1H),6.77(d,J＝8.6Hz,1H),6.36(t,J＝2.7Hz,1H),4.67–4.55(m,1H),3.41–3.36(m,4H),2.48–2.41(m,4H),2.39–2.29(m,4H),2.28–2.20(m,2H),2.16(s,3H),2.09–2.01(m,2H),1.87–1.72(m,10H),1.68–1.60(m,1H),1.60–1.50(m,2H)；MS:629[M+H] +.
实施例11.(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-异丙基苯基)氨 基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000039
实施例11的制备参考实施例6的制备路线的步骤1)至步骤5)，其中步骤1)中以等摩尔当量的溴代异丙烷替代1-碘代-2-甲基丙烷，步骤2)中以等摩尔当量的2-甲基-1-(甲基(哌啶-4-基)氨基)丙-2-醇替代N,N-二甲基哌嗪-4-胺。 1H NMR(600MHz,DMSO-d6)δ11.60(s,1H),11.21(s,1H),9.15–9.10(m,1H),8.72(s,1H),7.59–7.53(m,1H),7.49(t,J＝7.9Hz,1H),7.39(s,1H),7.38–7.36(m,1H),7.06(t,J＝7.6Hz,1H),6.93–6.91(m,1H),6.79(d,J＝8.5Hz,1H),6.37–6.35(m,1H),4.60–4.51(m,1H),3.97(s,1H),3.42–3.38(m,2H),3.29(s,2H),2.34(s,3H),2.31–2.26(m,2H),1.83(d,J＝13.5Hz,6H),1.78–1.70(m,2H),1.61–1.51(m,2H),1.29(d,J＝6.0Hz,6H),1.25–1.21(m,1H),1.09(s,6H)；MS:620[M+H] +.
实施例12.(2-((2-((3-异丙基-4-(4-((2-甲氧基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000040
实施例12的制备参考实施例6的制备路线的步骤1)至步骤5)，其中步骤1)中以等摩尔当量的溴代异丙烷替代1-碘代-2-甲基丙烷，步骤2)中以等摩尔当量的N-(2-甲氧基乙基)-N-甲基哌啶-4-胺替代N,N-二甲基哌嗪-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.60(s,1H),11.21(s,1H),9.17–9.10(m,1H),8.72(s,1H),7.61–7.52(m,1H),7.49(t,J＝7.9Hz,1H),7.41–7.35(m,2H),7.10–7.03(m,1H),6.94–6.90(m,1H),6.79(d,J＝8.3Hz,1H),6.38–6.34(m,1H),4.60–4.51(m,1H),3.44–3.35(m,5H),3.25(s,3H),2.60(t,J＝6.2Hz,2H),2.41–2.32(m,2H),2.24(s,3H),1.83(d,J＝13.5Hz,6H),1.78–1.73(m,2H),1.59–1.51(m,2H),1.29(d,J＝6.0Hz,6H)；MS:606[M+H] +.
实施例13.(2-((2-((4-(4-((环丙基甲基)(甲基)氨基)哌啶-1-基)-3-异丙基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000041
实施例13的制备参考实施例6的制备路线的步骤1)至步骤5)，其中步骤1)中以等摩尔当量的溴代异丙烷替代1-碘代-2-甲基丙烷，步骤2)中以等摩尔当量的N-(环丙基甲基)-N-甲基哌啶-4-胺替代N,N-二甲基哌嗪-4-胺。 1H NMR(400MHz,DMSO-d6)δ11.60(s,1H),11.21(s,1H),9.16–9.09(m,1H),8.72(s,1H),7.61–7.52(m,1H),7.49(t,J＝7.9Hz,1H),7.42–7.34(m,2H),7.06(t,J＝7.3Hz,1H),6.94–6.90(m,1H),6.79(d,J＝8.4Hz,1H),6.38–6.32(m,1H),4.60–4.49(m,1H),3.41–3.36(m,2H),2.47–2.44(m,2H),2.34–2.31(m,2H),2.28(s,3H),1.83(d,J＝13.5Hz,6H),1.78–1.73(m,2H),1.63–1.49(m,3H),1.29(d,J＝6.0Hz,6H),0.84–0.78(m,1H),0.48–0.43(m,2H),0.11–0.05(m,2H)；MS:602[M+H] +.
实施例14.(2-((2-((3-异丙基-4-(4-(4-异丙基哌嗪-1-基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000042
实施例14的制备参考实施例6的制备路线的步骤1)至步骤5)，其中步骤2)中以等摩尔当量的1-异丙基-4-(哌啶-4-基)哌嗪替代N,N-二甲基哌嗪-4-胺。 1HNMR(400MHz,DMSO-d 6)δ11.64(s,1H),11.31–11.21(m,1H),9.76–9.16(m,1H),9.14–9.09(m,1H),8.87(s,1H),7.64–7.48(m,3H),7.42(d,J＝8.6Hz,1H),7.12–7.05(m,1H),7.00–6.94(m,1H),6.40–6.36(m,1H),4.70–4.53(m,1H),3.54–3.44(m,4H),3.33–3.27(m,4H),3.09–2.88(m,3H),2.79–2.57(m,3H),2.06–1.92(m,2H),1.84(d,J＝13.5Hz,6H),1.78–1.59(m,2H),1.33(d,J＝6.0Hz,6H),1.28–1.16(m,6H)；MS:645[M+H] +.
实施例15.(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000043
步骤1)叔-丁基(1-(2-甲氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基甲酸酯的制备
将叔-丁基甲基(哌啶-4-基)氨基甲酸酯214毫克(1mmol)、1-氟-2-甲氧基-4-硝基苯171毫克(1mmol)置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，搅拌至反应完毕，加水、抽滤得产品329毫克，产率90％。MS:366[M+H] +.
步骤2)1-(2-甲氧基-4-硝基苯基)-N-甲基哌啶-4-胺的制备
将叔-丁基(1-(2-甲氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基甲酸酯329毫克(0.9mmol)置于反应瓶中，加入4毫升二氯甲烷和1毫升三氟乙酸，搅拌至反应完毕，蒸去溶剂，加入乙酸乙酯和碳酸钠水溶液萃取，有机相浓缩得产品239毫克，产率100％。MS:266[M+H] +.
步骤3)1-((1-(2-甲氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇的制备
将1-(2-甲氧基-4-硝基苯基)-N-甲基哌啶-4-胺239毫克(0.9mmol)，2,2-二甲基环氧乙烷72毫克(1mmol)置于反应瓶中，加入乙醇，加热搅拌至反应完毕，蒸去溶剂，柱层析得产品213毫克，产率70％。MS:338[M+H] +.
步骤4)1-((1-(4-氨基-2-甲氧基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇的制备
将1-((1-(2-甲氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇213毫克(0.63mmol)，置于反应瓶中，加入甲醇，雷尼镍200毫克，在氢气环境下搅拌至反应完毕，抽滤，浓缩得产品194毫克，产率100％。MS:308[M+H] +.
步骤5).(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
将1-((1-(4-氨基-2-甲氧基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇194毫克(0.63mmol)，(2-((2-氯-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧磷283毫克(0.63mmol)，三二亚苄基丙酮二钯91毫克(0.1mmol)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽58毫克(0.1mmol)以及碳酸铯325毫克(1mmol)置于反应瓶中，加入二氧六环，加热搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品227毫克，产率50％。MS:722[M+H] +.
步骤6).(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
将(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-甲氧基苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦227毫克(0.32mmol)，置于反应瓶中，加入二氯甲烷4毫升，三氟乙酸1毫升，搅拌至原料消失，蒸去溶剂，加入甲醇4毫升。饱和氢氧化钠水溶液0.5毫升，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品151毫克，产率80％。 1H NMR(400MHz,DMSO-d6)δ11.64(s,1H),11.25(s,1H),9.18–9.13(m,1H),8.79(s,1H),7.60–7.53(m,1H),7.51–7.45(m,2H),7.37–7.34(m,1H),7.09–7.03(m,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.80(d,J＝8.6Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),3.99(s,1H),3.78(s,3H),3.37–3.34(m,2H),3.33–3.30(m,2H),2.48–2.47(m,1H),2.35(t,3H),2.30(s,2H),1.83(d,J＝13.5Hz,6H),1.76–1.70(m,2H),1.63–1.52(m,2H),1.08(s,6H).MS:592[M+H] +.
实施例16.(2-((2-((4-(4-(乙基(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000044
步骤1)8-(2-异丙氧基-4-硝基苯基)-1,4-二氧-8-氮杂螺[4.5]癸烷的制备
将1-氟-2-异丙氧基-4-硝基苯199毫克(1mmol)、1,4-二氧-8-氮杂螺[4.5]癸烷143毫克(1mmol)置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，搅拌至反应完毕，加水、抽滤得产品290毫克，产率90％。MS:323[M+H] +.
步骤2)3-异丙氧基-4-(1,4-二氧-8-氮杂螺[4.5]癸-8-基)苯胺的制备
将8-(2-异丙氧基-4-硝基苯基)-1,4-二氧-8-氮杂螺[4.5]癸烷290毫克(0.9mmol)、置于反应瓶中，加入甲醇，雷尼镍200毫克，氢气环境下搅拌至反应完毕，抽滤、浓缩后得产品263 毫克，产率100％。MS:293[M+H] +.
步骤3).(2-((2-((3-异丙氧基-4-(1,4-二氧-8-氮杂螺[4.5]癸-8-基)苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
将3-异丙氧基-4-(1,4-二氧-8-氮杂螺[4.5]癸-8-基)苯胺263毫克(0.9mmol)，(2-((2-氯-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧磷404毫克(0.9mmol)，三二亚苄基丙酮二钯91毫克(0.1mmol)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽58毫克(0.1mmol)以及碳酸铯325毫克(1mmol)置于反应瓶中，加入二氧六环，加热搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品318毫克，产率50％。MS:707[M+H] +.
步骤4).1-(4-((4-((2-(二甲基磷酰基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-2-基)氨基)-2-异丙氧基苯基)哌啶-4-酮的制备
将(2-((2-((3-异丙氧基-4-(1,4-二氧-8-氮杂螺[4.5]癸-8-基)苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦318毫克(0.45mmol)，置于反应瓶中，加入三氟乙酸2毫升，水2毫升，搅拌至原料消失，蒸去溶剂，加入甲醇和饱和氢氧化钠水溶液，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品120毫克，产率50％。MS:533[M+H] +.
步骤5).(2-((2-((4-(4-(乙基(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
将1-(4-((4-((2-(二甲基磷酰基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-2-基)氨基)-2-异丙氧基苯基)哌啶-4-酮120毫克(0.225mmol)，置于反应瓶中，加入二氯甲烷，N-甲基乙胺13毫克(0.225mmol)以及三乙酰氧基硼氢化钠95毫克(0.45mmol)，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩，柱层析得产品110毫克，产率85％。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.24(s,1H),9.14(dd,J＝8.6,4.3Hz,1H),8.75(s,1H),7.61–7.47(m,2H),7.41–7.37(m,2H),7.09–7.04(m,1H),6.94–6.91(m,1H),6.80(d,J＝8.4Hz,1H),6.38–6.35(m,1H),4.59–4.53(m,1H),3.40(d,J＝11.1Hz,2H),2.65–2.52(m,4H),2.49–2.47(m,1H),2.27(s,3H),1.83(d,J＝13.5Hz,6H),1.79(s,2H),1.65–1.56(m,2H),1.30(d,J＝6.0Hz,6H),1.03(t,J＝7.0Hz,3H).MS:576[M+H] +.
实施例17.(2-((2-((3-乙氧基-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000045
步骤1)叔-丁基(1-(2-羟基-4-硝基苯基)哌啶-4-基)(甲基)氨基甲酸酯的制备
将叔-丁基甲基(哌啶-4-基)氨基甲酸酯214毫克(1mmol)、2-氟-5-硝基苯酚157毫克(1mmol)置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，搅拌至反应完毕，加水、抽滤得产品315毫克，产率90％。MS:352[M+H] +.
步骤2)叔-丁基(1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基甲酸酯的制备
将叔-丁基(1-(2-羟基-4-硝基苯基)哌啶-4-基)(甲基)氨基甲酸酯315毫克(0.9mmol)、溴乙烷107毫克(1mmol)置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，搅拌至反应完毕，加水、抽滤得产品307毫克，产率90％。MS:380[M+H] +.
步骤3)1-(2-乙氧基-4-硝基苯基)-N-甲基哌啶-4-胺的制备
将叔-丁基(1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基甲酸酯307毫克(0.81mmol)置于反应瓶中，加入4毫升二氯甲烷和1毫升三氟乙酸，搅拌至反应完毕，蒸去溶剂，加入乙酸乙酯和碳酸钠水溶液萃取，有机相浓缩得产品226毫克，产率100％。MS:280[M+H] +.
步骤4)1-((1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇的制备
将1-(2-乙氧基-4-硝基苯基)-N-甲基哌啶-4-胺226毫克(0.81mmol)，2,2-二甲基环氧乙烷72毫克(1mmol)置于反应瓶中，加入乙醇，加热搅拌至反应完毕，蒸去溶剂，柱层析得产品200毫克，产率70％。MS:352[M+H] +.
步骤5)1-((1-(4-氨基-2-乙氧基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇的制备
将1-((1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇200毫克(0.56mmol)，置于反应瓶中，加入甲醇，雷尼镍200毫克，在氢气环境下搅拌至反应完毕，抽滤，浓缩得产品180毫克，产率100％。MS:322[M+H] +.
步骤6)(2-((2-((3-乙氧基-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
将1-((1-(4-氨基-2-乙氧基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇180毫克(0.56mmol)，(2-((2-氯-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦252毫克(0.56mmol)，三二亚苄基丙酮二钯91毫克(0.1mmol)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽58毫克(0.1mmol)以及碳酸铯325毫克(1mmol)置于反应瓶中，加入二氧六环，加热搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品205毫克，产率50％。MS:736[M+H] +.
步骤7)(2-((2-((3-乙氧基-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
将(2-((2-((3-乙氧基-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦205毫克(0.28mmol)，置于反应瓶中，加入二氯甲烷4毫升，三氟乙酸1毫升，搅拌至原料消失，蒸去溶剂，加入甲醇4毫升。饱和氢氧化钠水溶液0.5毫升，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品131毫克，产率80％。。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.24(d,J＝2.3Hz,1H),9.17–9.12(m,1H),8.75(s,1H),7.60–7.54(m,1H),7.50–7.44(m,2H),7.36–7.31(m,1H),7.09–7.04(m,1H),6.94–6.92(m,1H),6.78(d,J＝8.6Hz,1H),6.37–6.34(m,1H),4.04–4.02(m,1H),4.00(q,J＝5.8Hz,2H),3.38(d,J＝11.3Hz,2H),2.49–2.43(m,3H),2.35(s,3H),2.30(s,2H),1.83(d,J＝13.5Hz,6H),1.74(d,J＝11.9Hz,2H),1.62–1.54(m,2H),1.37(t,J＝6.9Hz,3H),1.08(s,6H).MS:606[M+H] +.
实施例18.(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000046
实施例18的制备方法参考实施例17的制备步骤1)至步骤7)，其中步骤2)中以等摩尔当量的1-溴丙烷替代溴乙烷。 1H NMR(400MHz,DMSO-d6)δ11.63(s,1H),11.24(s,1H),9.14(dd,J＝8.6,4.3Hz,1H),8.75(s,1H),7.60–7.53(m,1H),7.48(t,J＝7.9Hz,1H),7.41(d,J＝2.3Hz,1H),7.36–7.32(m,1H),7.09–7.04(m,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.78(d,J＝8.6Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),3.99(s,1H),3.91(d,J＝6.3Hz,2H),3.41–3.36(m,2H),2.44(d,J＝15.4Hz,3H),2.34(s,3H),2.30(s,2H),1.83(d,J＝13.5Hz,6H),1.79–1.71(m,4H),1.63–1.53(m,2H),1.08(s,6H),1.04(t,J＝7.4Hz,3H).MS:620[M+H] +.
实施例19.(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-异丁氧基苯基)氨 基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000047
实施例19的制备方法参考实施例17的制备步骤1)至步骤7)，其中步骤2)中以等摩尔当量的1-溴-2-甲基丙烷替代溴乙烷。 1H NMR(400MHz,DMSO-d6)δ11.64(s,1H),11.23(s,1H),9.14(dd,J＝8.5,4.3Hz,1H),8.74(s,1H),7.60–7.53(m,1H),7.50–7.45(m,1H),7.40–7.34(m,2H),7.09–7.04(m,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.78(d,J＝8.4Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.00(s,1H),3.71(d,J＝6.2Hz,2H),3.39(d,J＝11.0Hz,2H),2.49–2.39(m,3H),2.34(s,3H),2.30(s,2H),2.10–2.02(m,1H),1.83(d,J＝13.5Hz,6H),1.74(d,J＝11.8Hz,2H),1.58(q,J＝10.7Hz,2H),1.08(s,6H),1.03(d,J＝6.7Hz,6H).MS:634[M+H] +.
实施例20.(2-((2-((3-丁氧基-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000048
实施例20的制备方法参考实施例17的制备步骤1)至步骤7)，其中步骤2)中以等摩尔当量的1-溴丁烷替代溴乙烷。 1H NMR(400MHz,DMSO-d6)δ11.63(s,1H),11.23(d,J＝2.6Hz,1H),9.14(dd,J＝8.6,4.3Hz,1H),8.74(s,1H),7.60–7.53(m,1H),7.50–7.45(m,1H),7.41(d,J＝2.3Hz,1H),7.35(dd,J＝8.5,2.3Hz,1H),7.08–7.04(m,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.78(d,J＝8.6Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.00(s,1H),3.96–3.91(m,2H),3.37(d,J＝11.6Hz,2H),2.49–2.40(m,3H),2.34(s,3H),2.30(s,2H),1.83(d,J＝13.5Hz,6H),1.77–1.70(m,4H),1.61–1.47(m,4H),1.08(s,6H),0.96(t,J＝7.4Hz,3H).MS:634[M+H] +.
实施例21.(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-(异戊氧基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000049
实施例21的制备方法参考实施例17的制备步骤1)至步骤7)，其中步骤2)中以等摩尔当量的1-溴-3-甲基丁烷替代溴乙烷。 1H NMR(400MHz,DMSO-d6)δ11.64(s,1H),11.24(s,1H),9.14(dd,J＝8.5,4.2Hz,1H),8.74(s,1H),7.60–7.53(m,1H),7.50–7.45(m,1H),7.42–7.34(m,2H),7.09–7.03(m,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.78(d,J＝8.5Hz,1H),6.37–6.35(m,1H),4.01–3.94(m,3H),3.40–3.33(m,5H),2.34(d,J＝4.5Hz,3H),2.29(s,2H),1.83(d,J＝13.5Hz,6H),1.77–1.70(m,2H),1.70–1.61(m,3H),1.60–1.52(m,2H),1.08(s,6H),0.95(d,J＝6.7Hz,6H).MS:648[M+H] +.
实施例22.(2-((2-((4-(4-(异丁基(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000050
参考实施例16制备方法的步骤1)至步骤5)，其中步骤5)中以等摩尔当量的N,2-二甲基丙-1-胺替代N-甲基乙胺。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.22(s,1H),9.14(dd,J＝8.6,4.3Hz,1H),8.74(s,1H),7.60–7.47(m,2H),7.41–7.36(m,2H),7.09–7.04(m,1H),6.92(dd,J＝3.5,2.2Hz,1H),6.80(d,J＝8.5Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.60–4.52(m,1H),3.43–3.36(m,3H),2.64–2.57(m,2H),2.21(s,3H),2.16(d,J＝7.2Hz,2H),1.83(d,J＝13.5Hz,6H),1.77–1.69(m,3H),1.62–1.53(m,2H),1.30(d,J＝6.0Hz,6H),0.86(d,J＝6.6Hz,6H).MS:604[M+H] +.
实施例23.(2-((2-((3-异丙氧基-4-(4-(甲基(丙基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000051
参考实施例16制备方法的步骤1)至步骤5)，其中步骤5)中以等摩尔当量的N-甲基丙-1-胺替代N-甲基乙胺。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23(d,J＝2.3Hz,1H),9.14(dd,J＝8.5,4.2Hz,1H),8.74(s,1H),87.60–7.47(m,2H),7.41–7.36(m,2H),7.09–7.04(m,1H),6.92(dd,J＝3.5,2.2Hz,1H),6.80(d,J＝8.5Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.61–4.53(m,1H),3.39(d,J＝11.4Hz,3H),2.46(d,J＝9.2Hz,2H),2.44–2.40(m,2H),2.23(s,3H),1.83(d,J＝13.5Hz,6H),1.80–1.74(m,2H),1.64–1.55(m,2H),1.47–1.39(m, 2H),1.30(d,J＝6.0Hz,6H),0.86(t,J＝7.3Hz,3H).MS:590[M+H] +.
实施例24.(2-((2-((4-(4-(二乙基氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000052
参考实施例16制备方法的步骤1)至步骤5)，其中步骤5)中以等摩尔当量的二乙胺替代N-甲基乙胺。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23(s,1H),9.14(dd,J＝8.6,4.3Hz,1H),8.74(s,1H),7.60–7.48(m,2H),7.41–7.37(m,2H),7.06(t,J＝7.6Hz,1H),6.94–6.91(m,1H),6.80(d,J＝9.2Hz,1H),6.37–6.35(m,1H),4.59–4.54(m,1H),3.39(d,J＝11.4Hz,2H),2.59–2.52(m,6H),2.47(s,1H),1.83(d,J＝13.5Hz,6H),1.75(d,J＝12.0Hz,2H),1.60–1.52(m,2H),1.29(d,J＝6.0Hz,6H),0.98(t,J＝7.0Hz,6H).MS:590[M+H] +.
实施例25.(2-((2-((4-(4-(二丙基氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000053
参考实施例16制备方法的步骤1)至步骤5)，其中步骤5)中以等摩尔当量的二丙基胺替代N-甲基乙胺。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.22(t,J＝2.1Hz,1H),9.14(dd,J＝8.5,4.3Hz,1H),8.74(s,1H),7.60–7.47(m,2H),7.38(d,J＝6.9Hz,2H),7.10–7.03(m,1H),6.92(dd,J＝3.5,2.2Hz,1H),6.79(d,J＝9.3Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.59–4.53(m,1H),3.42–3.37(m,2H),2.49–2.45(m,3H),2.43–2.38(m,4H),1.83(d,J＝13.5Hz,6H),1.72(d,J＝12.0Hz,2H),1.61–1.52(m,2H),1.43–1.35(m,4H),1.29(d,J＝6.0Hz,6H),0.86(t,J＝7.3Hz,6H).MS:618[M+H] +.
实施例26.(2-((2-((3-异丙氧基-4-(4-(吡咯烷-1-基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000054
参考实施例16制备方法的步骤1)至步骤5)，其中步骤5)中以等摩尔当量的吡咯烷替代N-甲基乙胺。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23(s,1H),9.14(dd,J＝8.4,4.3Hz,1H),8.74(s,1H),7.60–7.47(m,2H),7.39(d,J＝7.3Hz,2H),7.09–7.04(m,1H),6.92(dd,J＝3.5,2.2Hz,1H),6.80(d,J＝9.0Hz,1H),6.37–6.35(m,1H),4.59–4.53(m,1H),3.31(s,2H),2.57–2.52(m,7H),1.97–1.90(m,2H),1.83(d,J＝13.5Hz,6H),1.69(s,4H),1.60–1.49(m,2H),1.29(d,J＝6.0Hz,6H).MS:588[M+H] +.
实施例27.(2-((2-((4-([1,4'-联哌啶]-1'-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000055
参考实施例16制备方法的步骤1)至步骤5)，其中步骤5)中以等摩尔当量的哌啶替代N-甲基乙胺。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23(s,1H),9.14(dd,J＝8.5,4.2Hz,1H),8.74(s,1H),7.60–7.47(m,2H),7.38(d,J＝7.1Hz,2H),7.09–7.04(m,1H),6.92(dd,J＝3.4,2.2Hz,1H),6.79(d,J＝9.2Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.59–4.52(m,1H),3.42–3.37(m,2H),2.50–2.44(m,7H),1.83(d,J＝13.5Hz,6H),1.80–1.76(m,2H),1.63–1.55(m,2H),1.53–1.47(m,4H),1.40(d,J＝6.6Hz,2H),1.29(d,J＝6.0Hz,6H).MS:602[M+H] +.
实施例28.(2-((2-((3-环丁氧基-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000056
实施例28的制备方法参考实施例17的制备步骤1)至步骤7)，其中步骤2)中以等摩尔 当量的溴代环丁烷替代溴乙烷。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.21(s,1H),9.15–9.10(m,1H),8.75(s,1H),7.61–7.48(m,2H),7.40–7.36(m,1H),7.26(d,J＝2.4Hz,1H),7.08(d,J＝7.5Hz,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.77(d,J＝8.7Hz,1H),6.38–6.34(m,1H),4.65–4.58(m,1H),3.99(s,1H),3.42–3.35(m,5H),2.35(s,3H),2.30(s,2H),2.09–2.00(m,3H),1.83(d,J＝13.5Hz,6H),1.79–1.71(m,3H),1.69–1.52(m,4H),1.08(s,6H).MS:632[M+H] +.
实施例29.(2-((2-((4-(4-(环丁基(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000057
参考实施例16制备方法的步骤1)至步骤5)，其中步骤5)中以等摩尔当量的N-甲基环丁基胺替代N-甲基乙胺。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23(s,1H),9.14(dd,J＝8.3,4.2Hz,1H),8.74(s,1H),7.60–7.47(m,2H),7.39(dd,J＝4.6,2.4Hz,2H),7.06(dd,J＝8.4,6.5Hz,1H),6.92(dd,J＝3.5,2.2Hz,1H),6.79(d,J＝9.2Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.58–4.52(m,1H),3.39(d,J＝12.7Hz,3H),3.32–3.28(m,2H),3.16(t,J＝7.9Hz,1H),2.08(s,3H),2.00–1.94(m,2H),1.83(d,J＝13.5Hz,6H),1.79(s,2H),1.66–1.56(m,6H),1.29(d,J＝6.0Hz,6H).MS:602[M+H] +.
实施例30.(2-((2-((3-(环戊氧基)-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000058
实施例30的制备方法参考实施例17的制备步骤1)至步骤7)，其中步骤2)中以等摩尔当量的溴代环戊烷替代溴乙烷。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.22(s,1H),9.14(dd,J＝8.5,4.2Hz,1H),8.74(s,1H),7.60–7.47(m,2H),7.42–7.34(m,2H),7.09–7.04(m,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.77(d,J＝8.6Hz,1H),6.36(dd,J＝3.5,1.9Hz,1H),4.78(d,J＝5.8Hz,1H),4.01(s,1H),3.39–3.36(m,2H),3.32–3.29(m,2H),2.45–2.43(m,1H),2.35(s,3H),2.30(s,2H),1.94–1.86(m,2H),1.83(d,J＝13.5Hz,6H),1.79–1.71(m,5H), 1.67–1.50(m,5H),1.08(s,6H).MS:646[M+H] +.
实施例31.(2-((2-((4-(4-(乙基(2-羟基-2-甲基丙基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000059
步骤1)N-乙基-1-(2-异丙氧基-4-硝基苯基)哌啶-4-胺的制备
将乙胺45毫克(1mmol)，1-(2-异丙氧基-4-硝基苯基)哌啶-4-酮278毫克(1mmol)置于反应瓶中，加入二氯甲烷10毫升，加入三乙酰氧基硼氢化钠424毫克(2mmol)，搅拌至反应完毕，二氯甲烷和水萃取，有机相浓缩得产品308毫克，产率100％。MS:308[M+H] +.
步骤2)1-(乙基(1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)氨基)-2-甲基丙-2-醇的制备
将N-乙基-1-(2-异丙氧基-4-硝基苯基)哌啶-4-胺308毫克(1mmol)，2,2-二甲基环氧乙烷87毫克(1.2mmol)置于反应瓶中，加入乙醇，加热搅拌至反应完毕，蒸去溶剂，柱层析得产品256毫克，产率70％。MS:380[M+H] +.
步骤3)至步骤5)(2-((2-((4-(4-(乙基(2-羟基-2-甲基丙基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
步骤3)至步骤5)参考实施例17的制备方法中的步骤5)至步骤7)，起始物料以等摩尔当量的1-(乙基(1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)氨基)-2-甲基丙-2-醇替代1-((1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇。 1H NMR(400MHz,DMSO-d6)δ11.61(s,1H),11.22(s,1H),9.14(dd,J＝8.4,4.2Hz,1H),8.73(s,1H),7.60–7.53(m,1H),7.50(t,J＝8.0,8.0Hz,1H),7.40–7.37(m,2H),7.06(t,J＝7.3,7.3Hz,1H),6.93–6.91(m,1H),6.81–6.78(m,1H),6.37–6.35(m,1H),4.59–4.52(m,1H),3.40(d,J＝11.0Hz,2H),2.64–2.58(m,4H),2.47–2.43(m,2H),2.36–2.33(m,2H),1.83(d,J＝13.5Hz,6H),1.74(d,J＝11.8Hz,2H),1.60–1.53(m,2H),1.30(s,3H),1.29(s,3H),1.08(s,6H),0.99(t,J＝7.0,7.0Hz,3H).MS:634[M+H] +.
实施例32.(2-((2-((3-(环己氧基)-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000060
实施例32的制备方法参考实施例17的制备步骤1)至步骤7)，其中步骤2)中以等摩尔当量的溴代环己烷替代溴乙烷。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.21(s,1H),9.16–9.11(m,1H),8.72(s,1H),7.60–7.47(m,2H),7.42–7.34(m,2H),7.09–7.03(m,1H),6.93(dd,J＝3.5,2.2Hz,1H),6.79(d,J＝8.6Hz,1H),6.36(dd,J＝3.4,1.9Hz,1H),4.37–4.29(m,1H),4.02(s,1H),3.41(d,J＝11.1Hz,2H),2.58–2.52(m,2H),2.48–2.45(m,1H),2.35(s,3H),2.31(s,2H),1.93–1.86(m,2H),1.83(d,J＝13.5Hz,6H),1.79–1.71(m,4H),1.62–1.48(m,5H),1.39–1.31(m,3H),1.09(s,6H).MS:660[M+H] +.
实施例33.(2-((2-((4-(4-(氮杂环辛烷-1-基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
步骤1)1-(1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)氮杂环辛烷的制备
Figure PCTCN2019088316-appb-000061
将氮杂环辛烷113毫克(1mmol)，1-(2-异丙氧基-4-硝基苯基)哌啶-4-酮278毫克(1mmol)置于反应瓶中，加入二氯甲烷10毫升，加入三乙酰氧基硼氢化钠424毫克(2mmol)，搅拌至反应完毕，二氯甲烷和水萃取，有机相浓缩得产品300毫克，产率80％。MS:376[M+H] +.
步骤2)至步骤4)(2-((2-((4-(4-(氮杂环辛烷-1-基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000062
步骤2)至步骤4)参考实施例17的制备方法中的步骤5)至步骤7)，起始物料以等摩 尔当量的1-(1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)氮杂环辛烷替代1-((1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23–11.21(m,1H),9.16–9.12(m,1H),8.74(s,1H),7.60–7.53(m,1H),7.52–7.47(m,1H),7.40–7.36(m,2H),7.08–7.04(m,1H),6.93–6.91(m,1H),6.81–6.78(m,1H),6.37–6.35(m,1H),4.59–4.53(m,1H),3.40–3.36(m,2H),2.59(t,J＝5.4,5.4Hz,4H),2.47–2.43(m,1H),1.83(d,J＝13.5Hz,6H),1.77–1.72(m,2H),1.65–1.50(m,14H),1.30(s,3H),1.29(s,3H).MS:630[M+H] +.
实施例34.(2-((2-((4-(4-(((1-羟基环丙基)甲基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000063
步骤1)至步骤3)1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺的制备
1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺的制备方法参考实施例17的制备步骤1)至步骤3)，其中步骤2)中以等摩尔当量2-溴代丙烷替代溴代乙烷。MS:294[M+H] +.
步骤4)1-羟基-N-(1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)-N-甲基环丙烷-1-甲酰胺的制备
将1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺293毫克(1mmol)，1-羟基环丙烷-1- 羧酸102毫克(1mmol)，2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(HATU)456毫克(1.2mmol)置于反应瓶中，加入N,N-二异丙基乙胺(DIEA)258毫克(2mmol)，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品302毫克，产率80％。MS:378[M+H] +.
步骤5)1-(((1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)甲基)环丙烷-1-醇的制备
将1-羟基-N-(1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)-N-甲基环丙烷-1-甲酰胺302毫克(0.8mmol)置于反应瓶中，加入10毫升甲苯，2M的硼烷二甲硫醚四氢呋喃溶液0.6毫升(1.2mmol)，加热搅拌至反应完毕，加入甲醇淬灭，浓缩后柱层析得产品174毫克，产率60％。MS:364[M+H] +.
步骤6)1-(((1-(4-氨基-2-异丙氧基苯基)哌啶-4-基)(甲基)氨基)甲基)环丙烷-1-醇的制备
将1-(((1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)甲基)环丙烷-1-醇174毫克(0.48mmol)置于反应瓶中，加入10毫升甲醇，200毫克雷尼镍，在氢气环境下搅拌至反应完毕，抽滤，浓缩后柱层析得产品160毫克，产率60％。MS:334[M+H] +.
步骤7)(2-((2-((4-(4-(((1-羟基环丙基)甲基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
制备方法参考实施例17中的步骤6)，其中以等摩尔量的1-(((1-(4-氨基-2-异丙氧基苯基)哌啶-4-基)(甲基)氨基)甲基)环丙烷-1-醇替代1-((1-(4-氨基-2-乙氧基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇。MS:748[M+H] +.
步骤8)(2-((2-((4-(4-(((1-羟基环丙基)甲基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
制备方法同实施例17中的步骤7)。 1H NMR(400MHz,DMSO-d6)δ11.73(s,1H),11.65(s,1H),9.09–9.04(m,1H),7.62–7.51(m,3H),7.20–7.18(m,1H),7.06(t,J＝7.4,7.4Hz,1H),6.59–6.54(m,1H),6.44–6.42(m,1H),6.21–6.17(m,1H),6.10–6.04(m,1H),4.64(s,1H),4.48–4.41(m,1H),3.18–3.13(m,2H),3.03(s,4H),2.68–2.66(m,1H),2.40–2.32(m,4H),2.21(s,3H),1.83(d,J＝13.5Hz,6H),1.73–1.68(m,2H),1.51–1.44(m,2H),1.23(s,3H),1.21(s,3H).MS:618[M+H] +.
实施例35.(2-((2-((4-(4-(((1-羟基环丁基)甲基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000064
步骤1)1-(((1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)氨基)甲基)环丁-1-醇的制备
将1-(氨基甲基)环丁-1-醇101毫克(1mmol)，1-(2-异丙氧基-4-硝基苯基)哌啶-4-酮278毫克(1mmol)置于反应瓶中，加入二氯甲烷10毫升，加入三乙酰氧基硼氢化钠424毫克(2mmol)，搅拌至反应完毕，二氯甲烷和水萃取，有机相浓缩得产品364毫克，产率100％。MS:364[M+H] +.
步骤2)1-(((1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)甲基)环丁-1-醇的制备
将1-(((1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)氨基)甲基)环丁-1-醇364毫克(1mmol)，置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，随后加入碘甲烷142毫克(1mmol)，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩得产品321毫克，产率85％。MS:378[M+H] +.
步骤3)1-(((1-(4-氨基-2-异丙氧基苯基)哌啶-4-基)(甲基)氨基)甲基)环丁-1-醇的制备
参考实施例34的制备方法中的步骤6)，起始原料中以等摩尔当量的1-(((1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)甲基)环丁-1-醇替代1-(((1-(2-异丙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)甲基)环丙烷-1-醇。MS:348[M+H] +.
步骤4)(2-((2-((4-(4-(((1-羟基环丁基)甲基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
制备方法参考实施例17中的步骤6)，其中以等摩尔量的1-(((1-(4-氨基-2-异丙氧基苯基)哌啶-4-基)(甲基)氨基)甲基)环丁-1-醇替代1-((1-(4-氨基-2-乙氧基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇。MS:762[M+H] +.
步骤5)(2-((2-((4-(4-(((1-羟基环丁基)甲基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
制备方法同实施例17中的步骤7)。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.22(s,1H),9.16–9.12(m,1H),8.74(s,1H),7.60–7.54(m,1H),7.50(t,J＝7.9,7.9Hz,1H),7.40–7.36(m,2H),7.09–7.04(m,1H),6.94–6.91(m,1H),6.80(d,J＝8.4Hz,1H),6.37–6.35(m,1H),4.71(s,1H),4.59–4.53(m,1H),3.42–3.38(m,2H),2.49–2.45(m,5H),2.34(s,3H),2.05–1.99(m,2H),1.94–1.88(m,2H),1.83(d,J＝13.5Hz,6H),1.80–1.75(m,2H),1.66–1.55(m,3H),1.47–1.40(m,1H),1.31(s,3H),1.29(s,3H).MS:632[M+H] +.
实施例36.(2-((2-((4-(4-(环丙基(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
步骤1)N-环丙基-1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺的制备
Figure PCTCN2019088316-appb-000065
参考实施例33的制备方法的步骤1)，以等摩尔当量的N-甲基环丙烷基胺取代氮杂环辛烷。MS:334[M+H] +.
步骤2)至步骤4)(2-((2-((4-(4-(环丙基(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000066
步骤2)至步骤4)参考实施例17的制备方法中的步骤5)至步骤7)，起始物料以等摩尔当量的N-环丙基-1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺替代1-((1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23(t,J＝2.1,2.1Hz,1H),9.14(dd,J＝8.6,4.3Hz,1H),8.74(s,1H),7.60–7.53(m,1H),7.50(t,J＝7.8,7.8Hz,1H),7.40–7.37(m,2H),7.09–7.04(m,1H),6.93–6.92(m,1H),6.82–6.79(m,1H),6.37–6.35(m,1H),4.59–4.53(m,1H),3.41–3.37(m,2H),2.55–2.52(m,2H),2.47–2.44(m,1H),2.29(s,3H),1.89–1.86(m,2H),1.83(d,J＝13.5Hz,6H),1.81–1.78(m,1H),1.67–1.59(m,2H),1.30(s,3H),1.29(s,3H),0.49–0.45(m,2H),0.33–0.29(m,2H).MS:588[M+H] +.
实施例37.(2-((2-((4-(4-(((1-羟基环戊基)甲基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000067
实施例37的制备方法参考实施例35的制备方法的步骤1)至步骤5)，起始物料中以等摩尔当量的1-(氨基甲基)环戊-1-醇替代1-(氨基甲基)环丁-1-醇。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.22(s,1H),9.16–9.12(m,1H),8.74(s,1H),7.60–7.53(m,1H),7.50(t,J＝7.9,7.9Hz,1H),7.40–7.36(m,2H),7.06(t,J＝7.2,7.2Hz,1H),6.94–6.91(m,1H),6.80(d,J＝8.4Hz,1H),6.37–6.35(m,1H),4.59–4.52(m,1H),3.98(s,1H),3.40(d,J＝11.0Hz,2H),2.48–2.47(m,1H),2.46–2.45(m,2H),2.35(s,3H),1.83(d,J＝13.5Hz,6H),1.77–1.68(m,4H),1.62–1.46(m,10H),1.30(s,3H),1.29(s,3H).MS:646[M+H] +.
实施例38.(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-(三氟甲氧基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000068
步骤1)叔-丁基(1-(4-氨基-2-(三氟甲氧基)苯基)哌啶-4-基)(甲基)氨基甲酸酯的制备
将叔-丁基甲基(哌啶-4-基)氨基甲酸酯214毫克(1mmol)，4-溴-3-(三氟甲氧基)苯胺255毫克(1mmol)，三二亚苄基丙酮二钯91毫克(0.1mmol)，2-二-叔丁膦基-2',4',6'-三异丙基联苯45毫克(0.1mmol)以及碳酸铯650毫克(2mmol)置于反应瓶中，加入二氧六环，加热搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品195毫克，产率50％。MS:390[M+H] +.
步骤2)叔-丁基(1-(4-((4-((2-(二甲基磷酰)苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-2-基)氨基)-2-(三氟甲氧基)苯基)哌啶-4-基)(甲基)氨基甲酸酯的 制备
将叔-丁基(1-(4-氨基-2-(三氟甲氧基)苯基)哌啶-4-基)(甲基)氨基甲酸酯195毫克(1mmol)，(2-((2-氯-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦225毫克(0.5mmol)，三二亚苄基丙酮二钯91毫克(0.1mmol)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽58毫克(0.1mmol)以及碳酸铯325毫克(1mmol)置于反应瓶中，加入二氧六环，加热搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品200毫克，产率50％。MS:804[M+H] +.
步骤3)二甲基(2-((2-((4-(4-(甲基氨基)哌啶-1-基)-3-(三氟甲氧基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)氧化膦的制备
将叔-丁基(1-(4-((4-((2-(二甲基磷酰)苯基)氨基)-7-((2-(三甲基硅基)乙氧基)甲基)-7H-吡咯并[2,3-d]嘧啶-2-基)氨基)-2-(三氟甲氧基)苯基)哌啶-4-基)(甲基)氨基甲酸酯200毫克(0.25mmol)，置于反应瓶中，加入二氯甲烷4毫升，三氟乙酸1毫升，搅拌至原料消失，蒸去溶剂，加入甲醇4毫升。饱和氢氧化钠水溶液0.5毫升，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩、柱层析得产品114毫克，产率80％。MS:574[M+H] +.
步骤4)(2-((2-((4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)-3-(三氟甲氧基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
将二甲基(2-((2-((4-(4-(甲基氨基)哌啶-1-基)-3-(三氟甲氧基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)氧化膦114毫克(0.2mmol)，2,2-二甲基环氧乙烷15毫克(0.2mmol)置于反应瓶中，加入乙醇，加热搅拌至反应完毕，蒸去溶剂，柱层析得产品90毫克，产率70％。 1H NMR(400MHz,DMSO-d6)δ11.67(s,1H),11.32(t,J＝2.2Hz,1H),9.10(d,J＝11.0Hz,2H),7.92–7.84(m,1H),7.78–7.70(m,1H),7.61–7.54(m,1H),7.54–7.47(m,1H),7.13–7.03(m,2H),6.99–6.92(m,1H),6.40–6.35(m,1H),4.01(s,1H),3.25(d,J＝11.0Hz,2H),2.68–2.56(m,2H),2.48–2.39(m,1H),2.34(s,3H),2.29(s,2H),1.83(d,J＝13.5Hz,6H),1.80–1.71(m,2H),1.61–1.46(m,2H),1.08(s,6H).MS:646[M+H] +.
实施例39.(2-((2-((4-(4-(二甲基氨基)哌啶-1-基)-3-(三氟甲氧基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000069
实施例39的制备方法参考实施例38的制备步骤1)至步骤3)，其中步骤1)中以等摩尔当量的N,N-二甲基哌啶-4-胺替代叔-丁基甲基(哌啶-4-基)氨基甲酸酯。 1H NMR(400MHz,DMSO-d6)δ11.67(s,1H),11.33(d,J＝2.2Hz,1H),9.10(d,J＝5.7Hz,2H),7.91–7.86(m, 1H),7.76–7.70(m,1H),7.62–7.54(m,1H),7.54–7.48(m,1H),7.12–7.02(m,2H),6.99–6.91(m,1H),6.41–6.34(m,1H),3.24(d,J＝11.4Hz,2H),2.69–2.59(m,2H),2.22(s,6H),2.20–2.14(m,1H),1.91–1.76(m,8H),1.55–1.45(m,2H).MS:588[M+H] +.
实施例40.(2-((2-((4-(4-((2-氟乙基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
步骤1)1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺的制备
Figure PCTCN2019088316-appb-000070
参考实施例35的制备方法的步骤1)，以等摩尔当量的甲胺替代将1-(氨基甲基)环丁-1-醇。MS:294[M+H] +.
步骤2)N-(2-氟乙基)-1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺的制备
Figure PCTCN2019088316-appb-000071
将1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺294毫克(1mmol)，置于反应瓶中，加入2毫升DMF和碳酸钾138毫克(1mmol)，随后加入1-溴-2-氟乙烷126毫克(1mmol)，搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩得产品289毫克，产率85％。MS:340[M+H] +.
步骤3)至步骤5)(2-((2-((4-(4-((2-氟乙基)(甲基)氨基)哌啶-1-基)-3-异丙氧基苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000072
步骤3)至步骤5)参考实施例17的制备方法中的步骤5)至步骤7)，起始物料以等摩尔当量的N-(2-氟乙基)-1-(2-异丙氧基-4-硝基苯基)-N-甲基哌啶-4-胺替代1-((1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇。 1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.23(t,J＝2.2,2.2Hz,1H),9.14(dd,J＝8.4,4.2Hz,1H),8.74(s,1H),7.60–7.53(m,1H),7.52–7.47(m,1H),7.41–7.36(m,2H),7.09–7.04(m,1H),6.94–6.92(m,1H),6.80(d, J＝8.4Hz,1H),6.37–6.35(m,1H),4.59–4.53(m,2H),4.43(t,J＝5.2,5.2Hz,1H),3.39(d,J＝11.2Hz,2H),2.78(t,J＝5.2,5.2Hz,1H),2.71(t,J＝5.2,5.2Hz,1H),2.54–2.52(m,1H),2.48–2.40(m,2H),2.28(s,3H),1.83(d,J＝13.5Hz,6H),1.77(d,J＝11.5Hz,2H),1.61–1.51(m,2H),1.30(s,3H),1.29(s,3H).MS:594[M+H] +.
实施例41.(2-((2-((3-(2-氟乙氧基)-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
Figure PCTCN2019088316-appb-000073
步骤1)1-苄基-N-甲基哌啶-4-胺的制备
将1-苄基哌啶-4-酮378毫克(2mmol)，甲胺62毫克(2mmol)置于反应瓶中，加入二氯甲烷10毫升，加入三乙酰氧基硼氢化钠848毫克(4mmol)，搅拌至反应完毕，二氯甲烷和水萃取，有机相浓缩得产品348毫克，产率85％。MS:205[M+H] +.
步骤2)1-((1-苄基哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇的制备
将1-苄基-N-甲基哌啶-4-胺348毫克(1.7mmol)，2,2-二甲基环氧乙烷144毫克(2mmol)置于反应瓶中，加入乙醇，加热搅拌至反应完毕，蒸去溶剂，柱层析得产品328毫克，产率70％。MS:277[M+H] +.
步骤3)2-甲基-1-(甲基(哌啶-4-基)氨基)丙-2-醇的制备
将1-((1-苄基哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇328毫克(1.2mmol)置于反应瓶中，加入甲醇，钯碳30毫克，在氢气环境下搅拌至反应完毕，抽滤，蒸去溶剂，浓缩得产品224毫克，产率100％。MS:187[M+H] +.
步骤4)1-((1-(2-(2-氟乙氧基)-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇的制备
将2-氟-5-硝基苯酚157毫克(1mmol)，置于反应瓶中，加入2毫升DMF和碳酸钾276毫克(2mmol)，随后加入1-溴-2-氟乙烷126毫克(1mmol)，搅拌至原料消失后加入2-甲基-1-(甲基(哌啶-4-基)氨基)丙-2-醇187毫克(1mmol)，继续搅拌至反应完毕，乙酸乙酯和水萃取，有机相浓缩得产品222毫克，产率60％。MS:370[M+H] +.
步骤5)至步骤7)(2-((2-((3-(2-氟乙氧基)-4-(4-((2-羟基-2-甲基丙基)(甲基)氨基)哌啶-1-基)苯基)氨基)-7H-吡咯并[2,3-d]嘧啶-4-基)氨基)苯基)二甲基氧膦的制备
步骤5)至步骤7)参考实施例17的制备方法中的步骤5)至步骤7)，起始物料以等摩尔当量的1-((1-(2-(2-氟乙氧基)-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇替代1-((1-(2-乙氧基-4-硝基苯基)哌啶-4-基)(甲基)氨基)-2-甲基丙-2-醇。 1H NMR(400MHz,DMSO-d6)δ11.63(s,1H),11.25(s,1H),9.17–9.09(m,1H),8.79(s,1H),7.62–7.53(m,1H),7.53–7.41(m,2H),7.39–7.32(m,1H),7.10–7.02(m,1H),6.97–6.91(m,1H),6.83(d,J＝8.7Hz,1H),6.39–6.33(m,1H),4.87–4.79(m,1H),4.75–4.65(m,1H),4.30–4.23(m,1H),4.22–4.15(m,1H),3.41(d,J＝11.1Hz,2H),2.57–2.51(m,2H),2.50–2.46(m,2H),2.46–2.33(m,5H),1.90–1.67(m,8H),1.69–1.55(m,2H),1.11(s,6H).MS:624[M+H] +.
生物活性测试
主要的仪器、试剂和细胞
Figure PCTCN2019088316-appb-000074
表1.所用主要的仪器、试剂和细胞
二.试剂配制：
EDTA(0.5M pH8.0)溶液配制:准确称量14.612g EDTA粉末，加入超纯水后定容到100mL(若有不溶加热到37℃，用NaOH溶液调pH至8.0)
1×Kinase Assay Buffer：于试剂瓶中分别加入25mL HEPES溶液(1M)、190.175mg EGTA、5mL MgCl 2溶液(1M)、1mL DTT、50μL Tween-20，加超纯水定容到500mL(调pH到7.5)。
1×Detection Buffer:取1mL 10×Detection Buffer加入9mL水混匀。
4×终止液：将0.8mL上述EDTA(0.5M、pH 8.0)溶液、1mL 10×Detection Buffer及8.2mL超纯水混匀。
4×EGFR T790M激酶溶液:用1×Kinase Assay Buffer稀释激酶原液到浓度为0.2nM，混匀，冰上保存。
4×底物溶液：用1×Kinase Assay Buffer稀释底物ULight TM-PolyGT原液到400nM，混匀。
4×ATP溶液：用1×Kinase Assay Buffer稀释ATP原液到浓度为20μM，混匀。
4×检测液：用1×Detection Buffer稀释检测抗体Eu-W1024-labeled Anti-Phosphotyrosine Antibody(PT66)到浓度为8nM，混匀。
2×底物/ATP混合液：将4×底物溶液和4×ATP溶液1:1等量混匀(使用前配制)。
实验例1.小分子化合物抑制EGFR T790M激酶活性的测试，测试方法如下：
1)化合物的稀释
在96孔板a中，将浓度为10mM的化合物用DMSO溶液按3倍比例稀释，形成11个梯度，第12个梯度为纯DMSO溶液(作为阳性对照)；取一块新的96孔板b，将上述溶液用超纯水稀释25倍(DMSO浓度为4％)。
2)将化合物转盘到384孔板
将上述96孔板b中用超纯水稀释过的化合物溶液按照2复孔的标准转盘到384孔板相应的孔中。
3)加4×激酶溶液：用排枪取2.5μl上述4×激酶溶液加入到384孔板相应的反应孔中，混匀室温预反应5分钟。
4)加2×底物/ATP混合液:用排枪取5μl上述2×底物/ATP混合液到384孔板相应的反应孔中。
5)阴性对照：在384孔板中设置阴性对照孔，每孔加入2.5μl 4×底物、2.5μl 4×酶溶液、2.5μl 1×Kinase Assay Buffer和2.5μl含4％DMSO的超纯水。
6)离心混匀，避光室温反应2小时。
7)终止酶促反应：
吸取5μl上述4×终止液到384孔板相应孔，离心混匀，室温反应5分钟。
8)显色反应：
吸取5μl上述4×检测液加入到384孔板相应孔，离心混匀，室温反应1小时。
9)将384孔板放入读板仪，调取相应的程序检测信号。
10)IC 50分析：
孔读值＝10000*EU665值/EU615值
抑制率＝(阳性对照孔读值-实验孔读值)/(阳性对照孔读值-阴性对照孔读值)*100％
将药物浓度和相应抑制率输入GraphPad Prism 5处理可计算出相应的IC 50。
实验例2.小分子化合物抑制EGFR L858R-T790M-C797S激酶活性的测试，测试方法如下：
在体外组装的酶促反应中，加入不同浓度的待测化合物，以检测化合物对EGFR T790M- L858R-C797S酶促反应的抑制作用，通过计算EGFR T790M-L858R-C797S酶促反应抑制的IC 50来筛选有生化抑制活性的化合物。其测试的具体方法与上述实验例1的方法一致，仅需调整以下条件：1).反应中ATP溶液终浓度为1μM；2).底物ULight TM-PolyGT的终浓度为50nM；3).EGFR L858R-T790M-C797S激酶的终浓度为0.5nM。
表2列出了本申请中实施例化合物对EGFR T790M激酶和EGFR L858R-T790M-C797S酪氨酸激酶抑制活性的测定结果，其中A表示IC 50小于或等于5nM，B表示IC 50大于5nM但小于或等于50nM，C表示IC 50大于50nM,NT表示没有测试对应的激酶。
Figure PCTCN2019088316-appb-000075
Figure PCTCN2019088316-appb-000076
表2:本发明化合物对EGFR T790M和EGFR L858R-T790M-C797S激酶抑制活性测定结果
实验例3.
小分子化合物抑制Ba/F3 EGFR-L858R/T790M及Ba/F3 EGFR-L858R/T790M/C797S细胞增殖的测试，具体方法如下：
1)Ba/F3 EGFR-L858R/T790M及Ba/F3 EGFR-L858R/T790M/C797S细胞用RPMI 1640完全培养基(RPMI 1640基础培养基+10％FBS)培养。
2)将生长状态良好的细胞收集转移至15mL离心管中，以1000rpm离心4分钟。
3)弃去上清液，加入上述完全培养基，吹打均匀，取10μL细胞悬浮液和10μL 0.4％胎盼蓝混匀，用细胞计数仪进行计数，记录细胞数及存活率。
4)每孔接种80μL的细胞悬液到96孔板中(不同细胞接种细胞密度见表3)。
细胞名称 培养基 接种密度
Ba/F3 EGFR-L858R/T790M RPMI 1640+10％FBS 5000/孔
Ba/F3 EGFR-L858R/T790M/C797S RPMI 1640+10％FBS 5000/孔
表3:细胞密度
5)在96孔板a中，将浓度为10mM的化合物用DMSO溶液按3倍比例稀释，形成9个梯度，第10个梯度为纯DMSO溶液(作为阳性对照)；取一块新的96孔板b，将上述溶液用RPMI 1640完全培养基稀释80倍得到5×化合物溶液(DMSO浓度为1.25％)。
6)在实验孔中加入20μL上述用培养液稀释过的5×化合物溶液，混合摇匀。
7)在含5％CO 2的37℃培养箱中培养72小时后每孔加入10μL CCK-8试剂，培养2小时(可以根据颜色深浅来调节反应时间)；
8)在多功能读板机于450nm处读其OD值。
9)数据处理：细胞存活率(％)＝[(As-Ab)/(Ac-Ab)]*100％
As：实验孔(含有细胞的培养基、CCK-8、化合物)的OD值；
Ac：对照孔(含有细胞的培养基、CCK-8)的OD值；
Ab：空白孔(不含细胞和化合物的培养基、CCK-8)的OD值。
然后将数值导入Graphpad Prism5软件进行曲线拟合，计算IC 50。
表4列出了本发明中实施例化合物对Ba/F3 EGFR-L858R/T790M和Ba/F3 EGFR-L858R/T790M/C797S细胞的活性测定结果，其中A表示IC 50小于或等于50nM，B表示IC 50大于50nM但小于或等于100nM，C表示IC 50大于100nM但小于或等于200nM，D表示IC 50大于200nM。
Figure PCTCN2019088316-appb-000077
Figure PCTCN2019088316-appb-000078
表4.本发明部分化合物对细胞活性的测定结果
实验例4.
小分子化合物抑制ALK激酶活性的测试，测试方法如下：
使用Perkin Elmer公司的
Figure PCTCN2019088316-appb-000079
Ultra TR-FRET技术，通过检测反应中ALK激酶的底物磷酸化的水平来测定其活性。在LANCE Ultra酶活性分析中，Eu标记的特异性抗磷酸抗体用来检测Ulight-底物的磷酸化。当检测抗体与磷酸化的底物结合时，Eu和Ulight两个基团接近，在接受320或340nm波长的激光辐射后，Eu发射的能量转移到Ulight上，并激发一个665nm的光信号。光信号强度与反应中Ulight-底物的磷酸化水平呈正相关，从而能够定量测定激酶的活性。
酶检测实验使用Carna Biosciences提供的ALK激酶，Promega公司提供的ATP，Perkin Elmer公司的
Figure PCTCN2019088316-appb-000080
Ultra Ulight TM–Poly GT底物、
Figure PCTCN2019088316-appb-000081
Eu-W1024-anti-phosphotyrosine检测抗体、LANCE TM Detection Buffer、白色OptiPlate-384孔板、封板膜以及Envison多功能读板机进行实验并检测。激酶缓冲液、终止液、显色液的配制方法、化合物稀释的方法如下：
1.激酶缓冲液的配制：于试剂瓶中分别加入25mL HEPES溶液(1M)、190.175mg EGTA、5mL MgCl 2溶液(1M)、1mL DTT、50μL Tween-20，加超纯水定容到500mL(调pH到7.5)。
2.检测液的配制：
Figure PCTCN2019088316-appb-000082
Eu-W1024-anti-phosphotyrosine检测抗体用1×Detection Buffer配制为8nM的浓度。
3.终止液的配制：将0.8mL EDTA(0.5M、pH 8.0)水溶液、1mL 10×Detection Buffer及8.2mL超纯水混匀。
4. 4×中间浓度化合物的稀释过程：先将化合物在纯DMSO中溶解稀释至10mM，然后用DMSO进行3倍系列稀释11个浓度，每个浓度点再用超纯H 2O稀释25倍。
ALK激酶、ATP以及
Figure PCTCN2019088316-appb-000083
Ultra Ulight TM–Poly GT底物用激酶缓冲液分别配制为2.8nM、4μM、200nM的4×中间浓度，完整酶促反应体系包括上述2.5μL的ALK激酶、2.5μL的ATP、2.5μL的稀释后的4×中间浓度化合物以及2.5μL的
Figure PCTCN2019088316-appb-000084
Ultra Ulight TM–Poly GT底物。组装酶促反应后在室温条件下避光反应2小时。酶促反应结束后加入5μL终止液终止酶促反应，而后加入5μL的显色检测液反应1小时。用Envison多功能读板机调取相应程序进行读板。
数据处理过程如下：读值＝10000*EU665值/EU615值
抑制百分率＝(阳性对照孔读值-实验孔读值)/(阳性对照孔读值-阴性对照孔读值)]*100％。
将药物浓度和相应抑制率输入GraphPad Prism5处理计算出相应的IC50。
表5列出了本申请中部分实施例化合物对ALK激酶抑制活性的测定结果，其中A表示IC50小于或等于5nM，B表示IC50大于5nM但小于或等于50nM，C表示IC50大于50nM。
Figure PCTCN2019088316-appb-000085
Figure PCTCN2019088316-appb-000086
表5:本发明部分化合物对ALK激酶抑制活性测定结果
从蛋白质数据库(RCSB Protein Data Bank)中的晶体结构(5J7H)得知，Brigatinib化合物结构中的哌啶环在与ALK蛋白的相互作用中与其相连接的苯环形成了近于90度的二面角。然而，哌啶环中的氮原子与其相连接的苯环所形成的共轭体系使得哌啶环倾向于与其相连接的苯环形成近于0度的二面角。
Figure PCTCN2019088316-appb-000087
如上述化合物结构(A)所示，在苯环的邻位上引入取代集团(诸如甲基等)将有助于哌啶环与苯环之间的转置，使其化合物三维结构更有利于与ALK蛋白形成相互作用。因此，这种结构修饰很有可能会大大增加化合物(A)以及类似化合物对ALK蛋白的生物活性。鉴于ALK与EGFR在ATP结合域的三维结构的相似性，化合物(A)以及类似化合物对C797S突变的EGFR的生物活性也预计将大大提高。基于以上构思，发明人设计并合成了本申请的系列化合物。
实验数据表明，本发明的化合物在苯环(相对于与其相连的哌啶环)的邻位上引入取代基有效地增强了突变型EGFR的细胞活性，其部分化合物对EGFR T790M以及C797S突变细胞的抑制活性远远高于Brigatinib，有希望成为***EGFR突变介导的非小细胞肺癌候选药物化合物。
以上所述仅为本发明的较佳实施方式而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。

Claims (14)

  1. 一种式(I)所示化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，
    Figure PCTCN2019088316-appb-100001
    式(I)中，
    X为CH或N；
    Y为CH或N；
    R 1为-NHR 5，R 5为未被取代或被取代的芳基、杂芳基、环烷基、环烷基桥环结构、并环结构，
    R 5中取代的芳基、杂芳基、环烷基、环烷基桥环结构、并环结构中的取代基为-CF 3、-OCF 3、羟基、氰基、卤素、C 1-C 6烷基、C 3-C 5环烷基、C 1-C 6烷氧基、C 3-C 5环烷基氧基、-S(＝O) 2R 6、-C(＝O)R 6、-P(＝O)R 6R 7、-S(＝O) 2NR 6R 7，
    R 6和R 7分别独立地为-H、C 1-C 6烷基、C 3-C 6环烷基，
    所述并环结构选自芳环并5-6元杂芳环基、5-6元杂芳环并5-6元杂芳环基、芳环并5-6元环烷基、芳环并5-6元杂环基、5-6元杂芳环并5-6元环烷基或5-6元杂芳环并5-6元杂环基；
    R 2为-H、-CF 3、-CH 2CF 3、C 1-C 6烷基、C 3-C 6环烷基、C 3-C 4环烷基取代的C 1-C 2烷基、含一个氧原子的4-6元杂环基、或-(CH 2) mR 8，其中m为1、2、3整数，
    R 8为-OH、-CN、-C(O)NH 2、-S(＝O) 2CH 3、C 1-C 3烷氧基、C 1-C 3烷硫基；
    R 3和R 4分别独立地为-H、C 1-C 6烷基、C 3-C 6环烷基、含一个氮原子或含一个氧原子的4-6元杂环基或-(CH 2) nR 9,其中n为1、2、3整数，
    所述含一个氮原子或含一个氧原子的4-6元杂环为非取代或被C 1-C 3的烷基所取代，
    R 9为-OH、-CN、-C(O)NH 2、-S(＝O) 2CH 3、-NR'R”、C 1-C 3烷氧基、C 1-C 3烷硫基，
    R'、R”分别独立地为H或C 1-C 3的烷基，
    或者，R 3、R 4与其相连的氮原子构成4-6元杂环或6-9元螺环，所述杂环含1-2个选自N、O或S的杂原子或含基团-C(＝O)-或-S(＝O) 2-作为环成员，
    所述R 3、R 4与其相连的氮原子构成的4-6元杂环为未被取代或分别被1-2个选自卤素、氰基、羟基、氨基、C 1-C 3烷基、C 1-C 3烷氧基、卤代C 1-C 3烷基、氰基取代的C 1-C 3烷基、羟基取代的C 1-C 3烷基、C 1-C 3烷氧基取代的C 1-C 3烷基所取代，
    所述R 3、R 4与其相连的氮原子构成的6-9元螺环为包含一个氮原子的单螺环。
  2. 根据权利要求1所述的化合物、其异构体、水合物、溶剂化物、药学上可接受的 盐及前药，其中，R 1为-NHR 5，R 5选自以下基团：
    Figure PCTCN2019088316-appb-100002
    Q为N或CH，
    T为NH，O或S，
    R 10选自-H、-OH、-F、-Cl、-Br、-CN、-CF 3、-OCF 3、甲基、乙基、丙基、异丙基、环丙基、环丁基、甲氧基、乙氧基、丙氧基、异丙氧基、环丙基氧基、环丁基氧基、或以下基团：
    Figure PCTCN2019088316-appb-100003
    R 11为-H、甲基、乙基、丙基、异丙基，
    R 12为-H、-F、-Cl、-Br、羟基、氰基、三氟甲基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基。
  3. 根据权利要求1所述的化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，其中，R 2选自-H、-CF 3、-CH 2CF 3、甲基、乙基、丙基、异丙基、1-甲基丙基、2-甲基丙基、环丙基、环丁基、环戊基、环己基、环丙基甲基、环丙基乙基、环丁基甲基、环氧丙烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基。
  4. 根据权利要求1所述的化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，其中，R 2选自正丁基、正戊基、异戊基、新戊基、正己基、异己基。
  5. 根据权利要求1所述的化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，其中，R 3和R 4分别独立地选自-H、甲基、乙基、丙基、异丙基、环丙基、环丁基、环戊基、环氧丙烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基、甲氨基乙基、二甲氨基乙基、甲氨基丙基、二甲氨基丙基，或R 3、R 4与其相连的氮原子构成4-6元杂环或6-9元螺环，所述4-6元杂环为取代或非取代的杂环，
    所述R 3、R 4与其相连的氮原子构成的取代或非取代的4-6元杂环选自以下环结构：
    Figure PCTCN2019088316-appb-100004
    R 13选自–H、甲氨基、乙氨基、二甲氨基，
    R 14选自-H、甲基、乙基、丙基、异丙基、甲酰基、乙酰基或甲磺酰基，
    R 15和R 16分别独立地选自-H、-F、-CF 3、羟基、氨基、氰基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基、异丙氧基、氰基甲基、氰基乙基、甲氧基甲基、甲氧基乙基、甲氧基丙基、羟甲基、羟乙基、羟丙基，
    所述R 3、R 4与其相连的氮原子构成的6-9元螺环选自以下螺环结构：
    Figure PCTCN2019088316-appb-100005
  6. 根据权利要求1所述的化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，其中，R 3选自-H、甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基、环丙基、环丁基、环戊基、环氧丙烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基、甲氨基乙基、二甲氨基乙基、甲氨基丙基、二甲氨基丙基，
    R 4选自正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基。
  7. 一种式(I)所示化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药，
    Figure PCTCN2019088316-appb-100006
    式(I)中，
    X为CH或N；Y为CH或N；
    R 1为-NHR 5，R 5选自以下基团：
    Figure PCTCN2019088316-appb-100007
    Q为N或CH，
    T为NH，O或S，
    R 10选自-H、-OH、-F、-Cl、-Br、-CN、-CF 3、-OCF 3、甲基、乙基、丙基、异丙基、环丙基、环丁基、甲氧基、乙氧基、丙氧基、异丙氧基、环丙基氧基、环丁基氧基、或以下基团：
    Figure PCTCN2019088316-appb-100008
    R 11为-H、甲基、乙基、丙基、异丙基，
    R 12为-H、-F、-Cl、-Br、羟基、氰基、三氟甲基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基；
    R 2选自-H、-CF 3、-CH 2CF 3、甲基、乙基、丙基、异丙基、1-甲基丙基、2-甲基丙基、环丙基、环丁基、环戊基、环己基、环丙基甲基、环丙基乙基、环丁基甲基、氧杂环丁烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、甲硫基乙基、甲硫基丙基、甲氧基乙基、甲氧基丙基、乙氧基乙基、乙氧基丙基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基；
    R 3选自-H、甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基、环丙基、环丁基、环戊基、氧杂环丁烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基；
    R 4选自-(CH 2) nR 17,其中n为1、2、3整数，
    R 17为
    Figure PCTCN2019088316-appb-100009
    或者C 3-C 6环烷基，R 18为-OH、-CN、-C(O)NH 2、-S(＝O) 2CH 3、C 1-C 3烷氧基、C 1-C 3烷硫基。
  8. 一种式(I)所示化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，
    Figure PCTCN2019088316-appb-100010
    式(I)中，
    X为CH或N；
    Y为CH或N；
    R 1为-NHR 5，R 5选自以下基团：
    Figure PCTCN2019088316-appb-100011
    Q为N或CH，
    T为NH、O或S，
    R 10选自-H、-OH、-F、-Cl、-Br、-CN、-CF 3、-OCF 3、甲基、乙基、丙基、异丙基、环丙基、环丁基、甲氧基、乙氧基、丙氧基、异丙氧基、环丙基氧基、环丁基氧基、或以下基团：
    Figure PCTCN2019088316-appb-100012
    R 11为-H、甲基、乙基、丙基、异丙基，
    R 12为-H、-F、-Cl、-Br、羟基、氰基、三氟甲基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基；
    R 2为-H、C 1-C 6烷基、C 3-C 6环烷基、卤代C 1-C 6烷基；
    R 3和R 4分别独立地为-H、C 3-C 8环烷基、或由1至3个选自C 1-C 6烷氧基、羟基、卤素、C 3-C 6环烷基的取代基所取代或者非取代的C 1-C 9烷基，
    或者，R 3、R 4与其相连的氮原子构成4-8元杂环，所述杂环还含1-2个选自N、O或S的杂原子或含基团-C(＝O)-或-S(＝O) 2-作为环成员，
    所述R 3、R 4与其相连的氮原子构成的4-8元杂环为未被取代或被1-2个选自卤素、氰基、羟基、氨基、C 1-C 3烷基、C 1-C 3烷氧基、卤代C 1-C 3烷基、氰基取代的C 1-C 3烷基、羟基取代的C 1-C 3烷基、C 1-C 3烷氧基取代的C 1-C 3烷基所取代。
  9. 根据权利要求8所述的化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，其中，X为CH；Y为CH；
    R 1为
    Figure PCTCN2019088316-appb-100013
    R 2为-H、甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、异戊基、戊基、新戊基、己基、异己基、环丙基、环丁基、环戊基、环己基、三氟甲基、氟乙基；
    R 3和R 4分别独立地选自-H、甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基、正己基、异己基、环丙基、环丁基、环戊基、环己基、氧杂环丁烷-3-基、四氢呋喃-3-基、四氢吡喃-4-基、四氢吡喃-3-基、N-甲基哌啶-3-基、N-甲基哌啶-4-基、N-甲基吡咯烷-3-基、N-甲基氮(杂)环丁烷-3-基、甲硫基乙基、甲硫基 丙基、甲氧基甲基、甲氧基乙基、甲氧基丙基、乙氧基甲基、乙氧基乙基、乙氧基丙基、异丙氧基甲基、异丙氧基乙基、异丙氧基丙基、氨基乙酰基、氨基丙酰基、甲磺酰乙基、甲磺酰丙基、羟甲基、羟乙基、羟丙基、氰基甲基、氰基乙基、氰基丙基、甲氨基乙基、二甲氨基乙基、甲氨基丙基、二甲氨基丙基、环丙基甲基、环丙基乙基、三氟甲基、氟乙基、2-羟基-2-甲基丙基、3-羟基-3-甲基丁基、
    Figure PCTCN2019088316-appb-100014
    或R 3、R 4与其相连的氮原子构成4-8元杂环，所述4-8元杂环为取代或非取代的杂环，
    所述R 3、R 4与其相连的氮原子构成的取代或非取代的4-8元杂环选自以下环结构：
    Figure PCTCN2019088316-appb-100015
    R 13选自–H、甲氨基、乙氨基、二甲氨基，
    R 14选自-H、甲基、乙基、丙基、异丙基、甲酰基、乙酰基或甲磺酰基，
    R 15和R 16分别独立地选自-H、-F、-CF 3、羟基、氨基、氰基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基、异丙氧基、氰基甲基、氰基乙基、甲氧基甲基、甲氧基乙基、甲氧基丙基、羟甲基、羟乙基、羟丙基。
  10. 一种化合物、其异构体、水合物、溶剂化物、药学上可接受的盐及前药，所述化合物选自：
    Figure PCTCN2019088316-appb-100016
    Figure PCTCN2019088316-appb-100017
  11. 一种制备权利要求1-9所述化合物、其异构体、水合物、溶剂化物、其药学上可接受的盐及其前药的方法，包括以下步骤，
    Figure PCTCN2019088316-appb-100018
  12. 一种治疗与酪氨酸激酶EGFR、HER2或ALK突变或过表达相关疾病的药用组合物，其由权利要求1-10任一项所述的式(I)的化合物或其药学上可接受的盐或其水合物或其溶剂化物或其前药与药学上可接受的载体或赋形剂组成。
  13. 一种药用组合物:其中包含如权利要求1-10任一项所述的式(I)的化合物或其药学上可接受的盐、水合物、溶剂化物、或前药作为活性成分，一个或多个其它的治疗剂，以及一种或多种药学上可接受的载体或赋形剂。
  14. 权利要求1至10中任一项所述的化合物、其异构体、水合物、溶剂化物、药学上可接受的盐或前药在制备治疗与酪氨酸激酶EGFR、HER2或ALK突变或过表达相关的癌症及自身免疫疾病的药物中的应用，其中所述癌症及自身免疫疾病包括眼底疾病、干眼症、银屑病、白癜风、皮炎、斑秃、类风湿性关节炎、结肠炎、多重硬化、***性红斑狼疮、克罗恩病、动脉粥样化、肺纤维化、肝纤维化、骨髓纤维化、非小细胞肺癌、小细胞肺癌、乳腺癌、胰腺癌、神经胶质瘤、胶质母细胞瘤、卵巢癌、子***、结肠直肠癌、黑色素瘤、子宫内膜癌、***癌、膀胱癌、白血病、胃癌、肝癌、胃肠间质瘤、甲状腺癌、慢性粒细胞白血病、急性髓细胞性白血病、非霍奇金淋巴瘤、鼻咽癌、食道癌、脑瘤、B细胞和T细胞淋巴瘤、淋巴瘤、多发性骨髓瘤、胆道癌肉瘤、胆管癌中的任一种。
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