WO2020082817A1 - 肟基萘醌类化合物及其制备方法和用途 - Google Patents

肟基萘醌类化合物及其制备方法和用途 Download PDF

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WO2020082817A1
WO2020082817A1 PCT/CN2019/095516 CN2019095516W WO2020082817A1 WO 2020082817 A1 WO2020082817 A1 WO 2020082817A1 CN 2019095516 W CN2019095516 W CN 2019095516W WO 2020082817 A1 WO2020082817 A1 WO 2020082817A1
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optionally substituted
phenyl
amino
alkyl
dihydronaphthalen
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French (fr)
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王恒山
经孝腾
黄日镇
贾强
褚长虎
吕玉泉
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普济生物科技（台州）有限公司
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Publication of WO2020082817A1 publication Critical patent/WO2020082817A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/20—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C251/00—Compounds containing nitrogen atoms doubly-bound to a carbon skeleton
    • C07C251/32—Oximes
    • C07C251/34—Oximes with oxygen atoms of oxyimino groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals
    • C07C251/44—Oximes with oxygen atoms of oxyimino groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals with the carbon atom of at least one of the oxyimino groups being part of a ring other than a six-membered aromatic ring
    • C07C251/46—Quinone oximes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C381/00—Compounds containing carbon and sulfur and having functional groups not covered by groups C07C301/00 - C07C337/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D333/30—Hetero atoms other than halogen
    • C07D333/34—Sulfur atoms

Definitions

  • the present application belongs to the technical field of pharmaceutical compounds and pharmaceuticals, and specifically relates to oxime naphthoquinone compounds, and preparation methods and uses thereof.
  • STAT Signal transduction and transcription activator
  • IDO dioxygenase
  • STAT3 Signal transduction and transcription activator 3
  • STAT3 is the essential activator of tumor cells and immune cells in the tumor microenvironment, and it is the convergence point of many oncogenic signaling pathways.
  • Studies on different types of cancer have shown that STAT3 dysregulation is a major player in cancer cell growth, proliferation, survival, angiogenesis, metastasis, and invasion, and is usually associated with a more malignant tumor phenotype.
  • STAT3 can transduce signals from many oncogenic proteins and pathways, and is a vital activator of many important immunosuppressive genes.
  • activated STAT3 plays a role in suppressing the host's immune surveillance of cancer, which also promotes the development of tumors.
  • the tumor can inhibit the T cell immune response by activating STAT3-mediated IDO to catalyze the oxidation of tryptophan necessary for T cell proliferation to kynurenine, causing tumor immune escape.
  • maintaining the pluripotency of glioblastoma stem cells requires receptor signaling regulated by STAT3. Inhibiting the activation of STAT3 will lead to the growth inhibition, differentiation and apoptosis of cancer stem cells, indicating that STAT3 is the survival site of cancer stem cells (CSC) Required.
  • CSC cancer stem cells
  • IDO1 Indoleamine 2,3-dioxygenase 1
  • X Indoleamine 2,3-dioxygenase 1
  • IDO1 Indoleamine 2,3-dioxygenase 1
  • multiple Poor prognosis associated with cancer types Platten, M .; Wick, W .; Van Den Eynde, BJ Cancer Cer. Res. 2012, 72, 5435-5440; Uyttenhove, C .; Pilotte, L .; Théate, I .; Stroobant, V .; Colau, D .; Parmentier, N .; Boon, T .; Van Den Eynde, BJ Nat. Med.
  • IDO1-mediated immune tolerance is widely accepted as one of the most important mechanisms for tumor evolution to escape immune surveillance.
  • human tumors such as breast cancer, prostate cancer, lung cancer, colon cancer, as well as neuroblastoma and melanoma
  • persistent high expression of IDO1 is common (Munn, DH; Mellor, ALClin. Invest. 2007 , 117,1147-1154.
  • IDO1 is a heme enzyme that catalyzes the oxidation of L-tryptophan (L-Trp) to N-formylkynurenine (NFK) by binding to molecular oxygen and cleaving pyrrole ring substrates.
  • L-Trp L-tryptophan
  • NFK N-formylkynurenine
  • the crystal structure of human IDO1 has a binding pocket in the terminal heme site (pocket A), which is connected to the second pocket (pocket B) at the entrance of the active site ( UF; Awad, L .; Grosididier, A .; Larrieu, P .; Stroobant, V .; Colau, D .; Cerundolo, V .; Simpson, AJG; Vogel, P .; Van den Eynde, BJ; Zoete, VJMed .Chem. 2010,53,1172-1189).
  • TNF- ⁇ tumor necrosis factor alpha
  • the initial host's inflammatory response to the tumor may cause IDO1 to be induced a second time (Dunn, GP; Koebel, CM; Schreiber, RDNat. Rev. Immunol. 2006, 6, 836-848).
  • IDO1 expression is positively correlated with different tumor progression parameters and shortening the survival time of patients. All these evidences suggest that inhibiting IDO1 may enhance the efficacy of cancer treatment drugs.
  • IDO1 inhibitors have synergistic effects in combination with some anti-cancer drugs such as cyclophosphamide, doxorubicin, paclitaxel, and cisplatin (Hou, DY; Muller, AJ; Sharma, MD; Du Hadaway, J .; Banerjee, T .; Johnson, M .; Mellor, AL; Prendergast, GC Cancer Res. 2007, 67, 792-801).
  • IDO1 has become a potential target for cancer immunotherapy.
  • IDO1 inhibitors A small number of IDO1 inhibitors have entered clinical research, and 1-methyl-DL-tryptophan developed by NewLink Genetics is the first IDO1 inhibitor (Cady, SG; Sono, M. Cancer Res. 1991, 291, 326- 333).
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3, or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido.
  • the 5- to 10-membered heteroaryl group is pyridyl or thienyl.
  • the C 6 -C 10 aryl group is optionally substituted with a group selected from the group consisting of halogen, hydroxyl, mercapto, optionally substituted C 1 -C 6 alkyl, any Optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted Halogenated C 1 -C 6 alkoxy.
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, An optionally substituted C 1 -C 6 alkoxy group, and an optionally substituted halogenated C 1 -C 6 alkyl group.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3, or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido.
  • the 5- to 10-membered heteroaryl group is pyridyl or thienyl.
  • the C 6 -C 10 aryl group is optionally substituted with a group selected from the group consisting of halogen, hydroxyl, mercapto, optionally substituted C 1 -C 6 alkyl, any Optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted Halogenated C 1 -C 6 alkoxy.
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, An optionally substituted C 1 -C 6 alkoxy group, and an optionally substituted halogenated C 1 -C 6 alkyl group.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3 or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido; and
  • R 4 is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted with one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optional substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional An optionally substituted amino group, an optionally substituted hydroxy C 1 -C 6 alkyl group, an optionally substituted halo C 1 -C 6 alkyl group, and an optionally substituted halo C 1 -C 6 alkoxy group.
  • the C 6 -C 10 aryl group is phenyl
  • the 5- to 10-membered heteroaryl is pyridyl or thienyl
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, An optionally substituted C 1 -C 6 alkoxy group, and an optionally substituted halogenated C 1 -C 6 alkyl group.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen
  • the C 6 -C 10 aryl group is phenyl
  • the 5- to 10-membered heteroaryl is thienyl
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3, or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido.
  • the 5- to 10-membered heteroaryl group is pyridyl or thienyl.
  • the C 6 -C 10 aryl group is optionally substituted with a group selected from the group consisting of halogen, hydroxyl, mercapto, optionally substituted C 1 -C 6 alkyl, any Optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted Halogenated C 1 -C 6 alkoxy.
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, An optionally substituted C 1 -C 6 alkoxy group, and an optionally substituted halogenated C 1 -C 6 alkyl group.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen
  • step b) reacting the product obtained in step a) with C 2 O 2 Cl 2 ;
  • step b) reacting the product obtained in step b) with X-NH 2 ;
  • step d) reacting the product obtained in step c) with hydrazine
  • step d) The product obtained in step d) is reacted with 1,4-naphthoquinone, the benzene ring in the 1,4-naphthoquinone is optionally substituted with 1 to m R 2 and the 2-position carbon is optionally substituted with R 3 substitution to obtain the compound of formula (II);
  • the reaction conditions of step a) are refluxing in an acidic solution such as acetic acid or glacial acetic acid at 55-85 ° C, preferably at 70 ° C, for 10-14 hours, preferably 12 hours;
  • the molar ratio is about 1: 1.
  • the reaction conditions of step b) are reaction at 0-15 ° C, preferably at 0 ° C, in a haloalkane solution such as dichloromethane or chloroform for 10-14 hours, preferably 12 Hour; suitable amount of dimethylformamide (DMF) can be added as a catalyst; the molar ratio of reactant oxalyl chloride to another reactant is 2.5: 1 or more.
  • a haloalkane solution such as dichloromethane or chloroform for 10-14 hours, preferably 12 Hour
  • suitable amount of dimethylformamide (DMF) can be added as a catalyst
  • the molar ratio of reactant oxalyl chloride to another reactant is 2.5: 1 or more.
  • the reaction condition of step c) is a reaction at 0-15 ° C, preferably at 0 ° C, in a halogenated alkane solution such as dichloromethane or chloroform for 20-40 minutes, preferably 30 Minutes, in which the dripping rate is controlled at 1 drop per second; triethylamine can be added as an acid binding agent; the molar ratio of the reactants is about 1: 1.
  • the reaction condition of step d) is to react in an alcohol solution such as absolute ethanol at room temperature, and the reaction time is about 2.5-3.5h until no more solid precipitates;
  • the molar ratio of hydrazine hydrate to another reactant is 2.5: 1 or more.
  • the reaction condition of step e) is to react in a mixed solution of triethylamine, dimethylformamide and water at room temperature for 18-24 hours; the reactants 1,4-naphthoquinone and The molar ratio of the other reactant is about 1.5: 1.
  • the step f) is to react the compound of formula (II) with hydroxylamine hydrochloride, the molar ratio of the compound of formula (II) to hydroxylamine hydrochloride is about 1: 3; preferably, the reaction condition of step f) is 70-80 in such a reaction a compound °C anhydrous methanol or ethanol solution of alcohol 10 to 14 hours, preferably 12 hours, to obtain a formula (I), wherein X, R 1, n, R 2, m and R 3 As described above.
  • step g) the reaction conditions of step g); 2: step g) the molar ratio of the compound of formula (I) with ClSO 2 R 4 Reaction of formula (I) with ClSO 2 R 4 to about 1 To react in an inert gas such as argon at 0-15 ° C, preferably at 0 ° C in dichloromethane for 20-40 minutes, preferably 30 minutes, then add triethylamine dropwise and react for 10-20 minutes;
  • an inert gas such as argon at 0-15 ° C, preferably at 0 ° C in dichloromethane for 20-40 minutes, preferably 30 minutes, then add triethylamine dropwise and react for 10-20 minutes;
  • an inert gas such as argon at 0-15 ° C, preferably at 0 ° C in dichloromethane for 20-40 minutes, preferably 30 minutes, then add triethylamine dropwise and react for 10-20 minutes;
  • the step f ') is to react the compound of formula (II) with methoxyamine hydrochloride.
  • the reaction condition of step f') is refluxing in pyridine at 80 ° C for 2 hours to obtain the formula
  • a pharmaceutical composition comprising a compound active in the present application or a pharmaceutically acceptable salt thereof, or a compound obtained according to a method of preparing a compound of formula (I) and formula (III) Or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, excipients, or a combination thereof.
  • an active compound of the present application or a pharmaceutically acceptable salt thereof a compound obtained according to a method of preparing a compound of formula (I) and formula (III) or a pharmaceutically acceptable thereof
  • a compound of the present application or a pharmaceutically acceptable salt thereof a compound obtained according to a method of preparing a compound of formula (I) and formula (III) or a pharmaceutically acceptable salt thereof, Or the use of the pharmaceutical composition of the present application in the preparation of a medicament for inhibiting STAT3 and / or IDO1.
  • a compound of the present application or a pharmaceutically acceptable salt thereof a compound obtained according to a method of preparing a compound of formula (I) and formula (III) or a pharmaceutically acceptable salt thereof, Or or the use of the pharmaceutical composition of the present application in the preparation of a medicament for treating cancer.
  • the cancer is selected from colon cancer, ovarian cancer, liver cancer, bladder cancer, cervical cancer, and lung small cell cancer.
  • Figure 1 shows a Western blot of p-STAT3 (Y705), p-STAT3 (S727), total STAT3 and ⁇ -actin from whole cell lysates from SKOV3 cells treated with compound Iu.
  • Figure 2 shows a Western blot of STAT1, STAT5 and ⁇ -actin treated with compound Iu from whole cell lysates from SKOV3 cells.
  • Fig. 3 shows a histogram of compound Iu selectively inhibiting STAT3, wherein the activity of compound Iu on STAT1, STAT3, STAT4 and STAT5 is detected using the luciferase method.
  • Figure 4 shows a bar graph of compound Iu inhibiting DNA binding of STAT3 (ELISA assay results).
  • Figure 5 shows that compound Iu inhibits pSTAT3 nuclear translocation, stimulates SKOV3 cells with IL-6, and treats the migration of p-STAT3 into the nucleus with Iu (0.5 ⁇ M).
  • Figure 6 shows a wound healing experiment of cell migration with Iu (0.1 and 0.5 ⁇ M) in SKOV3 cancer cell line.
  • Figure 7 shows that compound Iu inhibits the invasion of SKOV3 cancer cell lines.
  • Figure 8 shows that compound Iu inhibits liver cancer growth in vivo.
  • FIG. 10 shows the surface plasmon resonance of the interaction between compound Iu and IDO1, which measures the binding between hIDO1 and compound Iu, and shows the SPR curve of IDO1 binding to Iu. It indicates the concentration of Iu injected on the surface of the biosensor chip immobilized with the IDO-1 protein. The measured K D was 0.08 ⁇ M.
  • Figure 11 shows that compound Iu inhibits melanoma growth in vivo.
  • This application provides dual-target selective inhibitors against STAT and IDO, especially dual-target selective inhibitors against STAT3 and IDO1.
  • the present application provides oxime naphthoquinone compounds, a preparation method thereof, and the compounds and pharmaceutical compositions thereof in the preparation of drugs for treating various tumors (especially ovarian cancer, colon cancer, and lung cancer, etc.) the use of.
  • this application designs and synthesizes a series of dual-target inhibitors against STAT3 and IDO1 for tumor immunotherapy.
  • UV-Vis spectroscopy analysis surface plasmon resonance analysis and other mechanism of action research, enzyme activity detection, Western Blot experiment, cell staining experiment, SKOV3 cell migration and invasion experiment and other cell activity tests, we found double action on STAT3 and IDO1 signaling pathway Target small molecule inhibitors. Specifically, it can inhibit SKOV3 cancer cell proliferation, migration and / or invasion.
  • the group When a group is described as “optionally substituted”, then the group may be unsubstituted or substituted with one or more of the substituents shown. Similarly, when a group is described as “unsubstituted or substituted”, if substituted, the substituent may be selected from one or more of the substituents shown.
  • the "optionally substituted” or “substituted” group may be substituted independently and independently with one or more groups selected from the group consisting of alkyl, alkenyl, alkyne Group, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclic, aralkyl, heteroaralkyl, (heteroalicyclic) alkyl, hydroxyl, hydrocarbyloxy, Aryloxy, acyl, mercapto, alkylthio, arylthio, cyano, halogen, thiocarbonyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thioamino Formyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyan
  • C a to C b refers to the number of carbon atoms in an alkyl group, alkenyl group, or alkynyl group, or the number of carbon atoms in an aryl group, heteroaryl group, where "a” and “b” are integers. That is, the alkyl, alkenyl, alkynyl, aryl, and heteroaryl groups may contain “a” to “b” carbon atoms, including “a” and “b” carbon atoms.
  • C 1 to C 4 alkyl refers to all alkyl groups having 1 to 4 carbons, ie CH 3- , CH 3 CH 2- , CH 3 CH 2 CH 2- , (CH 3 ) 2 CH-, CH 3 CH 2 CH 2 CH 2- , CH 3 CH 2 CH (CH 3 )-and (CH 3 ) 3 C-. If the "a” and “b" of alkyl, alkenyl, alkynyl, aryl, and heteroaryl groups are not specified, the broadest scope is assumed to be described in these definitions.
  • Alkyl refers to a straight or branched hydrocarbon chain that contains a fully saturated (no double or triple bond) hydrocarbon group.
  • An alkyl group can have 1 to 20 carbon atoms, and whenever it appears herein, a numerical range such as “1 to 20” refers to each integer in a given range; for example, "1 to 20 carbon atoms” means that Contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, 10 carbon atoms , 11 carbon atoms, 12 carbon atoms, 13 carbon atoms, 14 carbon atoms, 15 carbon atoms, 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, 19 carbon atoms or 20 carbon atoms Of alkyl.
  • the alkyl group may also be a medium-sized alkyl group having 1 to 10 carbon atoms.
  • the alkyl group may also be a lower alkyl group having 1 to 6 carbon atoms.
  • the alkyl group of the compound may be designated as "C 1 -C 6 alkyl group” or a similar name.
  • C 1 -C 4 alkyl means one to four carbon atoms in the alkyl chain, that is, the alkyl chain is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl , Sec-butyl and tert-butyl.
  • Typical alkyl groups include, but are in no way limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, and hexyl.
  • the alkyl group may be substituted or unsubstituted.
  • Alkenyl refers to a hydrocarbon group containing one or more double bonds in a linear or branched hydrocarbon chain.
  • the alkenyl group can be unsubstituted or substituted.
  • An alkenyl group can have 1 to 20 carbon atoms, and whenever it appears herein, a numerical range such as “1 to 20" refers to each integer in the given range; for example, “1 to 20 carbon atoms” refers to Contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, 10 carbon atoms , 11 carbon atoms, 12 carbon atoms, 13 carbon atoms, 14 carbon atoms, 15 carbon atoms, 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, 19 carbon atoms or 20 carbon atoms Of alkenyl.
  • Alkynyl refers to a hydrocarbon group containing one or more triple bonds in a linear or branched hydrocarbon chain.
  • the alkynyl group may be unsubstituted or substituted.
  • An alkynyl group may have 1 to 20 carbon atoms, and whenever it appears herein, a numerical range such as “1 to 20" refers to each integer in the given range; for example, “1 to 20 carbon atoms” refers to Contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, 10 carbon atoms , 11 carbon atoms, 12 carbon atoms, 13 carbon atoms, 14 carbon atoms, 15 carbon atoms, 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, 19 carbon atoms or 20 carbon atoms Of alkynyl.
  • Aryl refers to a carbocyclic (all carbon) monocyclic or polycyclic aromatic ring system (including fused ring systems in which two carbon rings share a chemical bond) with a completely delocalized ⁇ -electron system throughout all rings .
  • the number of carbon atoms in the aryl group can vary.
  • the aryl group may be a C 6 -C 14 aryl group, a C 6 -C 10 aryl group, or a C 6 aryl group.
  • Examples of aryl groups include, but are not limited to benzene, naphthalene, and azulene.
  • the aryl group may be substituted or unsubstituted.
  • Heteroaryl refers to containing one or more (eg, 1, 2, 3, 4, 5, or 6) heteroatoms (ie, elements other than carbon include but are not limited to nitrogen, oxygen And sulfur) monocyclic or polycyclic aromatic ring systems (ring systems with completely delocalized ⁇ -electron systems).
  • the number of atoms in the heteroaryl ring can vary.
  • the heteroaryl group may contain 4 to 14 atoms in the ring, 5 to 10 atoms in the ring, or 5 to 6 atoms in the ring.
  • heteroaryl includes fused ring systems in which two rings, for example at least one aryl ring and at least one heteroaryl ring or at least two heteroaryl rings, share at least one chemical bond.
  • heteroaryl rings include, but are not limited to, furan, furan, thiophene, benzothiophene, phthalazine, pyrrole, oxazole, benzoxazole, 1,2,3-oxadiazole, 1,2,4- Oxadiazole, thiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole, benzothiazole, imidazole, benzimidazole, indole, indazole, pyrazole, benzopyrazole, Isoxazole, benzisoxazole, isothiazole, triazole, benzotriazole, thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine
  • Alkoxy refers to the formula -OR, where R is an alkyl group as defined herein.
  • R is an alkyl group as defined herein.
  • Non-limiting lists of alkoxy groups are methoxy, ethoxy, n-propoxy, 1-methylethoxy (isopropoxy), n-butoxy, isobutoxy, sec-butoxy , Tert-butoxy.
  • the alkoxy group may be substituted or unsubstituted.
  • acyl refers to hydrogen, alkyl, alkenyl, alkynyl, or aryl as a substituent connected through a carbonyl group. Examples include formyl, acetyl, propionyl, benzoyl and acryl. The acyl group may be substituted or unsubstituted.
  • Haloalkyl refers to an alkyl group in which one or more hydrogen atoms are replaced with halogen (eg, monohaloalkyl, dihaloalkyl, and trihaloalkyl). Such groups include, but are not limited to, chloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, 1-chloro-2-fluoromethyl, and 2-fluoroisobutyl.
  • halogen eg, monohaloalkyl, dihaloalkyl, and trihaloalkyl.
  • Such groups include, but are not limited to, chloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, 1-chloro-2-fluoromethyl, and 2-fluoroisobutyl.
  • the haloalkyl group may be substituted or unsubstituted.
  • Halohydrocarbyloxy refers to a hydrocarbyloxy group in which one or more hydrogen atoms are replaced by halogen (eg, monohalohydrocarbyloxy, dihalohydrocarbyloxy, and trihalohydrocarbyloxy).
  • halogen eg, monohalohydrocarbyloxy, dihalohydrocarbyloxy, and trihalohydrocarbyloxy.
  • Such groups include, but are not limited to, chloromethoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, 1-chloro-2-fluoromethoxy, and 2-fluoroisobutoxy.
  • the halogenated hydrocarbyloxy group may be substituted or unsubstituted.
  • Arylthio refers to RS-, where R is aryl, such as but not limited to phenyl.
  • the arylthio group may be substituted or unsubstituted.
  • hydrocarbon thio refers to the "-SR" group, where R can be hydrogen, alkyl (the “hydrocarbon thio" at this time is “alkylthio"), alkenyl, alkynyl, cycloalkyl , Cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclic, aralkyl, (heteroaryl) alkyl or (heteroalicyclic) alkyl.
  • the hydrocarbon thio group may be substituted or unsubstituted.
  • the sulfinyl group may be substituted or unsubstituted.
  • the “sulfonyl” group refers to a “SO 2 R” group, where R is the same as R in the hydrocarbon thio group.
  • the sulfonyl group can be substituted or unsubstituted.
  • the O-carboxyl group may be substituted or unsubstituted.
  • the thiocarbonyl group may be substituted or unsubstituted.
  • trihalomethanesulfonyl refers to the "X 3 CSO 2- " group, where each X is halogen.
  • trihalomethanesulfinamido refers to the "X 3 CS (O) 2 N (R A )-" group, where each X is halogen and R A is hydrogen, alkyl, alkenyl, Alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclic, aralkyl, (heteroaryl) alkyl or (heteroalicyclic) alkyl.
  • amino refers to the -NH 2 group.
  • hydroxyl refers to the -OH group.
  • the "cyano” group refers to the "-CN” group.
  • the “isocyanate” group refers to the "-NCO” group.
  • the "thiocyanate” group refers to the "-CNS” group.
  • the “isothiocyanate” group refers to the "-NCS” group.
  • the “mercapto” group refers to the "-SH” group.
  • the "S-sulfonylamino” group refers to a "-SO 2 N (R A R B )" group, where R A and R B can independently be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl , Cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclic, aralkyl, (heteroaryl) alkyl or (heteroalicyclic) alkyl.
  • the S-sulfonylamino group may be substituted or unsubstituted.
  • N-sulfonamido group refers to "RSO 2 N (R A) -" group, wherein R A and R may be independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl Group, cycloalkynyl group, aryl group, heteroaryl group, heteroalicyclic group, aralkyl group, (heteroaryl) alkyl group or (heteroalicyclic group) alkyl group.
  • the N-sulfonylamino group may be substituted or unsubstituted.
  • the O-carbamoyl group may be substituted or unsubstituted.
  • the N-carbamoyl group may be substituted or unsubstituted.
  • the O-thiocarbamoyl group may be substituted or unsubstituted.
  • the N-thiocarbamoyl group may be substituted or unsubstituted.
  • the C-amido group may be substituted or unsubstituted.
  • the N-amido group may be substituted or unsubstituted.
  • halogen atom refers to any of the radio-stable atoms in column 7 of the periodic table, such as fluorine, chlorine, bromine, and iodine.
  • pharmaceutically acceptable salt refers to a salt of a compound that does not cause significant irritation to the organism to which it is administered and does not destroy the biological activity and properties of the compound.
  • the salt is an acid addition salt of the compound.
  • Pharmaceutical salts can be obtained by reacting compounds with inorganic acids such as hydrohalic acid (eg, hydrochloric acid or hydrobromic acid), sulfuric acid, nitric acid, and phosphoric acid.
  • Pharmaceutical salts can be obtained by reacting compounds with organic acids such as aliphatic or aromatic carboxylic acids or sulfonic acids, such as formic acid, acetic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, tobacco Acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid or naphthalenesulfonic acid.
  • organic acids such as aliphatic or aromatic carboxylic acids or sulfonic acids, such as formic acid, acetic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, tobacco Acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid or naphthalenesulfonic acid.
  • Pharmaceutical salts can also be obtained by reacting compounds with bases to form salts, such as ammonium salts, alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, such as dicyclohexylamine, N Methyl-D-glucosamine, tris (hydroxymethyl) methylamine, C 1 -C 7 alkylamines, cyclohexylamine, triethanolamine, ethylenediamine, and organic base salts such as arginine and lysine Amino acid salt of amino acid.
  • salts such as ammonium salts, alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, such as dicyclohexylamine, N Methyl-D-glucosamine, tris (hydroxymethyl) methylamine, C 1 -C 7 alkylamines, cyclohexylamine, triethanolamine, ethylenediamine, and organic base
  • each center may independently be R-configuration or S-configuration or a mixture thereof. Therefore, the compounds provided herein can be enantiomerically pure, enantiomerically enriched, racemic mixtures, diastereomerically pure, diastereomerically enriched or stereo Isomer mixture. Furthermore, it should be understood that in any compound described herein that produces one or more double bonds that can be defined as geometric isomers of E or Z, each double bond can independently be E or Z, or a mixture thereof.
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3, or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido.
  • the 5- to 10-membered heteroaryl is pyridyl or thienyl.
  • the C 6 -C 10 aryl group is optionally substituted with a group selected from the group consisting of halogen, hydroxyl, mercapto, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy.
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkoxy, and optionally substituted halogenated C 1 -C 6 alkyl.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3, or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido.
  • the 5- to 10-membered heteroaryl is pyridyl or thienyl.
  • the C 6 -C 10 aryl group is optionally substituted with a group selected from the group consisting of halogen, hydroxyl, mercapto, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy.
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkoxy, and optionally substituted halogenated C 1 -C 6 alkyl.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3 or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido; and
  • R 4 is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted with one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optional substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional An optionally substituted amino group, an optionally substituted hydroxy C 1 -C 6 alkyl group, an optionally substituted halo C 1 -C 6 alkyl group, and an optionally substituted halo C 1 -C 6 alkoxy group.
  • the C 6 -C 10 aryl group is phenyl
  • the 5- to 10-membered heteroaryl is pyridyl or thienyl
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkoxy, and optionally substituted halogenated C 1 -C 6 alkyl.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen
  • the C 6 -C 10 aryl group is phenyl
  • the 5- to 10-membered heteroaryl is thienyl
  • X is C 6 -C 10 aryl or 5- to 10-membered heteroaryl optionally substituted by one or more groups selected from halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, Optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optional Substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted halo C 1 -C 6 alkoxy;
  • Each R 1 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, and optionally substituted halogenated C 1 -C 6 alkoxy;
  • n is an integer selected from 1, 2, 3, 4 or 5;
  • Each R 2 is independently selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1- C 6 alkyl, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl , O-thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido;
  • n is an integer selected from 1, 2, 3, or 4;
  • R 3 is selected from hydrogen, halogen, hydroxyl, mercapto, optionally substituted hydrocarbon thio, optionally substituted C 1 -C 6 alkyl, optionally substituted C 1 -C 6 alkenyl, optionally substituted C 1 -C 6 alkynyl, optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl Group, optionally substituted halogenated C 1 -C 6 alkoxy, sulfinyl, sulfonyl, S-sulfonylamino, N-sulfonylamino, O-carbamoyl, N-carbamoyl, O- Thiocarbamoyl, N-thiocarbamoyl, C-amido, and N-amido.
  • the 5- to 10-membered heteroaryl group is pyridyl or thienyl.
  • the C 6 -C 10 aryl group is optionally substituted with a group selected from the group consisting of halogen, hydroxyl, mercapto, optionally substituted C 1 -C 6 alkyl, any Optionally substituted C 1 -C 6 alkoxy, optionally substituted amino, optionally substituted hydroxy C 1 -C 6 alkyl, optionally substituted halo C 1 -C 6 alkyl, and optionally substituted Halogenated C 1 -C 6 alkoxy.
  • the C 6 -C 10 aryl group is phenyl, which is optionally substituted with a group selected from halogen, optionally substituted C 1 -C 6 alkyl, An optionally substituted C 1 -C 6 alkoxy group, and an optionally substituted halogenated C 1 -C 6 alkyl group.
  • each R 1 is hydrogen
  • each R 2 is hydrogen
  • R 3 is hydrogen.
  • X is optionally substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 groups.
  • the specific experimental method of this application is based on phthalic anhydride and L-phenylalanine to synthesize the oxime naphthoquinone derivatives represented by formula (I), formula (II), formula (III) and (IV) .
  • the reactants are phthalic anhydride and L-phenylalanine (the phenyl group in L-phenylalanine is optionally substituted with 1 to n R 1 ), reaction conditions: 70 ° C, reflux in acetic acid 12h;
  • reaction conditions triethylamine, DCM, 0 ° C; drop The speed is controlled at 1 drop per second, and the reaction time is about 30 min.
  • reaction conditions absolute ethanol, room temperature; reaction time is about 2.5-3.5h.
  • a 2-amino acid amide-oxime naphthoquinone compound (compounds Ia-Iz), which is prepared through step f): the product of step e) above is reacted with hydroxylamine hydrochloride (H 3 NO ⁇ HCl) The reaction conditions are at 70 °C -80 °C, refluxing in absolute ethanol for 12h.
  • a 2-amino acid amide-hydroxime ester-based naphthoquinone compound (compounds IIIk-1, IIIq, IIIr, IIIk-2), which is prepared through step g): the product of step f) above ClSO 2 R 4 (R 4 is optionally substituted C6-C10 aryl or 5- to 10-membered heteroaryl, such as 4-tert-butylbenzenesulfonyl chloride) reaction, reaction conditions: DCM, argon, triethyl Amine, water, 0 °C, reaction time: about 40min.
  • the active compound of the present application When used as a medicine, it can be used directly or in the form of a pharmaceutical composition.
  • the pharmaceutical composition contains 0.1-99%, preferably 0.5-90% of the active compound of the present application, and the rest is a pharmaceutically acceptable pharmaceutically acceptable carrier that is non-toxic and inert to humans and animals.
  • the proportion of active compound contained in the pharmaceutical composition is 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99%.
  • Pharmaceutically acceptable carriers are one or more solid, semi-solid, and liquid diluents, fillers, and excipients for pharmaceutical products.
  • the pharmaceutical composition of the present application is used in the form of dosage per unit weight.
  • the medicines used in the present application may adopt the dosage forms commonly used in the art, such as ointments, tablets, pills, suppository emulsions, infusion solutions and injection solutions.
  • These dosage forms are prepared according to well-known methods using traditional additives and excipients.
  • the medicament prepared in this way can be administered by local, parenteral, oral and other routes as required.
  • the application amount of the active compound of the present application may vary according to the route of administration, the age, weight of the patient, the type and severity of the disease being treated, etc.
  • the daily dose may be 0.01-10 mg / kg body weight, preferably 0.1-5 mg / kg body weight. It can be administered one or more times.
  • the sources and specifications of the reagents used in the examples of the present application are as follows.
  • 1,4-naphthoquinone (specification: 25g), hydroxylamine hydrochloride (specification: 25g), 4-t-butylbenzenesulfonyl chloride (specification: 25g), L-phenylalanine (specification: 500g), methoxyamine Hydrochloride (specification: 25g), phthalic anhydride (specification: 500g), oxalyl chloride (specification: 500ml) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd .;
  • Triethylamine (specification: 500ml), dichloromethane (specification: 500ml), N, N-dimethylformamide (specification: 500ml), glacial acetic acid (specification: 500ml), hydrochloric acid (specification: 500ml), anhydrous Sodium sulfate (specification: 500g), absolute ethanol (specification: 500ml), ethyl acetate (specification: 5L), petroleum ether (specification: 5L), pyridine (specification: 500ml) were purchased from Guangdong Shantou Xilong Science Co., Ltd. ;
  • Hydrazine hydrate (specification: 500ml) was purchased from Sinopharm Group Chemical Reagent Co., Ltd .;
  • the above reagents are of analytical grade.
  • reaction mixture was spin-dried under reduced pressure, dissolved in dichloromethane, transferred to a constant-pressure burette, and added dropwise with 1 mL of triethylamine and NH 2 -X in an ice bath (the amount of the reactant was 1: 1) Add to the round bottom flask to control the drop rate of one drop per second. After the drop addition is complete, continue to stir. After the gas has evaporated, it is dried under reduced pressure.
  • NH 2 -X is, for example, 4- (trifluoromethyl) aniline, 2-fluoroaniline, aniline, 3-fluoroaniline, 2-methoxyaniline, 3,5-dimethylaniline, 2-toluidine, 4 -Chloroaniline, 4-bromoaniline, 3-bromo-4-fluoroaniline, 3- (trifluoromethyl) aniline, 4-fluoroaniline, 3-toluidine, 2-chloroaniline, 3-chloroaniline, 2- Bromoaniline, 3-bromoaniline, 3-methoxyaniline, 4-methoxyaniline, 4-toluidine, 3,5-dimethoxyaniline, 3-fluoro-4-bromoaniline, 3,5- Difluoroaniline, 2-fluoro-4-methylaniline, 2,5-dimethoxyaniline, 3-chloro-4-methylaniline, 2-methyl-4-bromoaniline.
  • the 1,4-naphthoquinone-2-aminoacylated substituted aniline compound synthesized above and methoxyamine hydrochloride were added to a round-bottom flask containing 25 mL of pyridine at a material ratio of 1:10 to control
  • the reaction was carried out under reflux at 80 ° C for 2h, rotary drying under reduced pressure, purification by silica gel column with eluent petroleum ether: ethyl acetate (6: 1), and the product obtained by rotary drying was a yellow-green solid.
  • the resulting product has the following specific structural formula and parameter characterization.
  • PBS buffer solution Take PBS powder, first dissolve it in 800 mL of deionized water, and then add 200 mL of deionized water to bring the volume to 1 L (10 mmol / L). It was autoclaved before the experiment and the PBS buffer after sterilization was stored at 4 ° C.
  • MTT stain methylthithiyltetrazolium, also known as thiazole blue
  • PBS phosphate buffer
  • Seed plate During the logarithmic growth of cells, after washing twice with PBS, digest with 0.25% trypsin, add medium to stop the digestion, and carefully pipet to obtain a single cell suspension. After counting, the cell suspension was diluted with an appropriate amount of 10% medium and seeded in 96-well plates, 180 ⁇ L per well, and the number of cells per well was 2 ⁇ 10 4 to 4 ⁇ 10 4 .
  • sample addition In a 96-well plate with different cancer cells, add 20 ⁇ L of the sample to be tested (sample concentration is 200 ⁇ M) to each well, and the final sample concentration is 20 ⁇ M for the initial screening. After screening, the samples with better results were selected for different concentration gradients and then sieved to calculate the IC 50. Each group had 9 duplicate wells. Each well of the control group was put into 20 ⁇ L of medium containing 10% DMSO and incubated for 48 hours. After adding the sample for 48 hours, each well was filled with 10 ⁇ L of MTT liquid with a concentration of 5 mg / mL and incubated for another 4 hours in a CO 2 incubator.
  • Doxorubicin (DOX) was used as a positive control, and the specific test results are shown in Table 1.
  • doxorubicin ** The structure of doxorubicin (DOX) is as follows:
  • Having one or more electron-withdrawing groups on the terminal aryl group can increase the anti-tumor activity in vitro.
  • a substituent that has certain spatial electronic properties to enhance lipophilicity has a strong ability to penetrate cell membranes, which can cause enhanced anti-proliferative effects.
  • the antitumor activity of Compound Iu against Hct-116, SKOV-3 and A549 was 0.037 ⁇ 0.01 ⁇ M, 0.028 ⁇ 0.01 ⁇ M, 0.033 ⁇ 0.01 ⁇ M and 0.012 ⁇ 0.01 ⁇ M, It is better than doxorubicin (0.27 ⁇ 0.08 ⁇ M, 0.16 ⁇ 0.17 ⁇ M, 0.074 ⁇ 0.05 ⁇ M and 0.069 ⁇ 0.02 ⁇ M, respectively), especially for human colon cancer cell Hct-116 and ovarian cancer cell SKOV3.
  • Run the gel after half an hour at 50V (run the concentrated gel), increase to 100V and run for 1.5 hours. Separate all the electrophoresis markers and stop when the indicator moves to the bottom of the electrophoresis tank.
  • transfer membrane buffer put medium-thick filter paper first, then put thin filter paper, and then put NC membrane in transfer membrane buffer. Remove the glue, cut the glue to the size of the filter paper, and place it horizontally on the NC membrane, taking care not to create bubbles. Then put thin filter paper on the glue, the top layer is thick filter paper, pay attention to drive out bubbles. Use tweezers to separate the uppermost thick filter paper from the lowermost medium thick filter paper to avoid scorching the NC membrane. Gently put the treated filter paper into a semi-dry film transfer instrument and transfer the film for 1.5 hours.
  • the film is soaked with 1 ⁇ Ponceau red dye solution for five minutes. After the second washing, a band appears, and the glue is cut according to the instructions of Marker.
  • the STAT family contains different subtype proteins such as STAT1, STAT2, STAT3, STAT4, STAT5 and STAT6.
  • STAT3 is often activated in tumor cells.
  • the expression of other subtype proteins was detected by Western blotting.
  • the results in Figure 2 indicate that compound Iu has no effect on the expression of STAT1 and STAT5.
  • the activities of STAT1, STAT3, STAT4 and STAT5 were detected using the luciferase method.
  • Luciferase assay cells were seeded in 24-well plates and transiently transfected with STAT3 reporter plasmid 4 ⁇ M67 pTATA TK-Luc and Renilla luciferase control reporter plasmids. Then, the cells were incubated with the complete medium for 24 hours after incubation with Compound Iu, the cells were lysed with 100 ⁇ L of cell lysis buffer in each well, and the supernatant was collected by centrifugation. Take 25 ⁇ L of supernatant and measure the luciferase activity of STAT3 by a microplate reader using a dual luciferase assay kit. The results are shown in Figure 3.
  • Enzyme-linked immunosorbent assay (ELISA) was used for detection.
  • Enzyme-linked immunosorbent assay 30 ⁇ L of complete binding buffer and 20 ⁇ L of STAT3 recombinant protein were added to each well, and the plate was gently stirred at room temperature for 1 hour.
  • 20 L of complete lysis buffer without STAT3 recombinant protein was used.
  • the wells were washed 3 times with 200 ⁇ L of 1X wash buffer and incubated with 100 uL of 1X HRP conjugated antibody (1: 1000 dilution) for 1 hour without stirring.
  • the cells were treated with different concentrations of compound Iu, and the samples were subjected to ELISA assay. As shown in FIG. 4, compared with the sample without compound treatment, compound Iu inhibited STAT3-DNA binding activity in a concentration-dependent manner, so it was proved that compound Iu inhibited the activation of STAT3 by blocking STAT3 and DNA.
  • SKOV3 cells were seeded on sterile slides and grown for 24 hours. The next day the cells were treated with compound Iu for 24 hours. For nuclear translocation experiments, after overnight serum-free, SKOV3 cells were pretreated with compound Iu for 2 hours, and then IL-6 (interleukin-6, interleukin 6) was added for 30 minutes. After treatment, cells were washed with cold phosphate buffered saline (PBS) buffer and fixed with cold methanol for 15 minutes at room temperature. After washing twice with pre-chilled PBS buffer, permeabilize the cells with X-100 containing 0.3% Triton at room temperature and block with 5% normal goat serum in PBS buffer for at least 1 hour.
  • PBS cold phosphate buffered saline
  • the cells were then incubated with phosphorylated STAT3 or STAT3 polyclonal rabbit antibody and overnight at 4 ° C. After overnight incubation, the cells were washed with PBS buffer containing 0.1% Tween-20. The cells were incubated with FITC fluorescent anti-rabbit secondary antibody for 1 hour at room temperature. After washing the cells, it was performed with DAPI, and then image analysis was performed by fluorescence microscope. The results are shown in Figure 5, and the experimental results are analyzed as follows.
  • SKOV3 cells were cultured in DMEM medium containing 10% FBS and the cells were seeded in 6-well plates. After the cells are overgrown, use a 10uL pipette tip to scratch and wash once with PBS to remove non-adherent cells. The culture plate was placed under an inverted microscope for observation and imaging, and the result served as a negative control without treatment. Fresh medium was replaced, cells were treated with compound Iu, and the cells were placed in an incubator and incubated for 24 hours. After incubation, observe and record the cell migration under the microscope. The experimental results are shown in Figure 6, and the experimental results are analyzed below.
  • the migration test was conducted in a 24-well intrusion cell (8.0 mm) polycarbonate membrane. First, add 600 ⁇ L of complete medium or treatment factor to the lower chamber of the invasion chamber. Next, 200 ⁇ L of DMEM suspension containing 2 ⁇ 10 5 / ml SKOV3 cells without FBS was inoculated into the upper chamber of the invasion chamber, and the cell migration filter was inserted into the lower chamber. Incubate at 37 ° C for 24 hours. Then, the SKOV3 cells on the upper side of the filter were removed, the migrated cells were fixed with 500 ⁇ L of 4% paraformaldehyde for 20 minutes, and the fixed cells were stained with hematoxylin for 3 minutes. Count the number of migrated cells and take pictures under a fluorescence microscope. The experimental results are shown in Figure 7. The experimental results are analyzed as follows.
  • Compound Iu was administered intravenously at a dose of 10 mg / kg or 20 mg / kg, which significantly inhibited tumor growth, resulting in 53.4% and 59.0% reduction in tumor growth compared to the negative control group (FIG. 8B).
  • Iu also reduced the tumor volume ( Figure 8C). It is worth noting that Compound 8u was well tolerated during the experiment and did not cause a significant weight loss in nude mice ( Figure 8D).
  • the human IDO1 enzyme activity test was performed according to the instructions provided by the reagent manufacturer (Tojo, S .; Kohno, T .; Tanaka, T .; Kamioka, S .; Ota, Y .; Ishii, T .; Kamimoto, K .; Asano, S. Med. Chem. Lett. 2014, 5, 1119-1123).
  • the IDO inhibitor IDO5L was used as a positive control.
  • the compounds synthesized in this application were tested for their inhibitory activity against human IDO1 (hIDO1) (human indoleamine 2,3-dioxygenase 1), and the IDO1 inhibitor IDO5L was used as a positive control.
  • hIDO1 human indoleamine 2,3-dioxygenase 1
  • IDO1 inhibitor IDO5L was used as a positive control.
  • the UV-visible absorption spectrum is measured on a Cary 1E UV-Visible spectrophotometer with a scanning range of 200-700 nm.
  • a Haake F3 water bath heating device was used in conjunction with a spectrophotometer.
  • the measurement uses 1 mL of a sample phosphate buffer solution (PBS) aqueous solution containing 10 ⁇ M of purified IDO1 and 25 ⁇ M of compound Iu.
  • PBS sample phosphate buffer solution
  • the experimental results are shown in Figure 9, and the results are analyzed as follows.
  • IDO1 has a heme cofactor active site, which enables the characterization of the interaction between the inhibitor and IDO.
  • the binding of the compound to heme iron can cause changes in the absorption of light, so it can be directly inferred that it binds to this binding site.
  • Heme is a porphyrin ring with iron coordination in the center. According to the oxidation and coordination state of iron, the maximum absorption wavelength of the ultraviolet-visible spectrum is about 400 nm. The iron binding of the inhibitor to heme will change the wavelength of its maximum absorbed light.
  • the absorption spectrum of the heme group is highly sensitive to the change in polarity around the heme after ligand / substrate binding, which changes the spectral characteristics of heme. Therefore, changes in the UV-visible spectrum caused by the interaction of IDO1 and ligands can be used to evaluate the binding of compounds to IDO1.
  • UV-Vis spectroscopy was used to detect the interaction between compound Iu and IDO1.
  • the absorption spectrum of IDO1 iron appears a Soret peak at 403 nm, which is consistent with previous literature reports (Yi-Hui Peng, Shau-Hua Ueng, Chen-Tso Tseng, et al, J. Med. Chem .2016,59,282-293).
  • the absorbed light is transferred to 413 nm, which proves that compound Iu binds to IDO1 and chelate with heme (see FIG. 9).
  • SPR Surface plasmon resonance
  • a GE Biacore T200 optical biosensor was used to measure surface plasmon resonance (SPR). As shown in Figure 10, within the selected concentration range, the response unit value (RU) is proportional to the concentration of the compound. According to the calculation of the software provided by the testing instrument, the equilibrium dissociation constant (KD) between compound Iu and IDO1 is 0.02 ⁇ M, which fully proves the binding affinity of compound Iu with the target IDO1 protein.

Abstract

本申请属于药物化合物和药物技术领域，涉及肟基萘醌类化合物及其制备方法和用途，具体涉及式(I)的化合物，该化合物能够作为STAT3和IDO1的双靶点选择性抑制剂，其用于治疗卵巢癌、结肠癌及肺癌等

Description

肟基萘醌类化合物及其制备方法和用途 技术领域
本申请属于药物化合物和药物技术领域，具体而言，涉及肟基萘醌类化合物及其制备方法和用途。
背景技术
恶性肿瘤是由癌细胞逃逸人类天然免疫系统而处于无限度增殖所导致，已成为现代人的主要致死性疾病。世界卫生组织(WHO)在《全球癌症报告2014》中称，全球每年新增癌症患者人数达1500万人，其中多种肿瘤(尤其是卵巢癌、结肠癌及肺癌等)是常见的高发性癌症。其治疗有效率低，实体瘤难以治疗，复发率在各种癌症中均处高位。由于这些肿瘤细胞可变异且逃逸免疫系统，导致化疗药物不具有高选择性，造成传统药物治疗失败，是目前抗肿瘤药临床中面临的重大挑战。因此，需要寻找新型高效低副作用的抗肿瘤制剂，尤其是能够选择性抑制肿瘤细胞脱分化无限扩增和针对免疫调节新机制的多靶点的新制剂。
信号传导与转录激活子(STAT)和双加氧酶(IDO)参与人体免疫调节与炎症以及胚胎发育和生长相关的生理过程，与多种癌症发生和恶性化有关，是恶性肿瘤治疗的重要靶点。STAT和IDO作为免疫检查点对调控免疫系统对肿瘤细胞的有效识别具有关键作用。
信号转导和转录激活因子3(STAT3)是肿瘤细胞和肿瘤微环境中免疫细胞本质的激活因子，它是许多致癌信号传导途径的收敛点。针对不同类型的癌症研究表明STAT3失调是癌细胞生长、增殖、存活、血管生成、转移和侵袭的主要参与者，并且通常与更恶性的肿瘤表型相关。STAT3能够转导来自众多致癌蛋白和通路的信号，而且是许多重要的免疫抑制基因至关重要的激活剂。而且，最近的研究证实，在不同的分子水平上，活化的STAT3在抑制宿主对癌症的免疫监视中起作用，这也促进了肿瘤的发展。肿瘤能通过激活STAT3介导的IDO催化氧化T细胞增殖必需的色氨酸转变为犬尿氨酸而抑制T细胞免疫应答，引起肿瘤免疫逃逸。此外，维持胶质母细胞瘤干细胞的多能性需要通过STAT3调控的受体信号传导，抑制STAT3的活化将导致癌症干细胞的生长抑制、分化以及凋亡，表明STAT3是癌症干细胞(CSC)存活所必需的。由此可见，STAT3已经成为癌症治疗潜在的药物靶标，开发STAT3抑制剂是有效缓解癌症耐药和迁移的可行方法。
目前，抑制STAT3活性的药物设计策略主要包括两种方法。一种是抑制其上游激酶，从而阻止STAT3的磷酸化进程。然而，由于这些激酶靶向多个下游蛋白质，当其他途径下调时可能产生不利的影响。另一个是直接抑制STAT3的 活性。与前一种方法相比，这种策略能最小地干扰正常细胞的功能，降低副作用，因而更具吸引力。迄今为止，已有多个STAT3抑制剂处于不同临床研究阶段，其中STA-21是第一个非肽的小分子STAT3抑制剂。STA-21是四角霉素的脱氧产物，具有醌的骨架结构。而随后发现的STAT3抑制剂LLL-3、LLL-12及LY-5均包含醌骨架。
吲哚胺2,3-双加氧酶1(IDO1)，其催化犬尿氨酸途径的初始和限速步骤，由肿瘤细胞表达以逃避潜在有效的免疫应答，并且高表达的IDO1与多种癌症类型的不良预后相关(Platten,M.；Wick,W.；Van den Eynde,B.J.Cancer Res.2012,72,5435-5440；Uyttenhove,C.；Pilotte,L.；Théate,I.；Stroobant,V.；Colau,D.；Parmentier,N.；Boon,T.；Van den Eynde,B.J.Nat.Med.2003,9,1269-1274；Théate,I.；van Baren,N.；Pilotte,L.；Moulin,P.；Larrieu,P.；Renauld,J.-C.；Hervé,C.；Gutierrez-Roelens,I.；Marbaix,E.；Sempoux,C.；Van den Eynde,B.J.Cancer Immunol.Res.2015,3,161-172；Godin-Ethier,J.；Hanafi,L.-A.；Piccirillo,C.A.Clin.Cancer Res.2011,17,6985-6991)。
目前，IDO1介导的免疫耐受性被广泛接受为肿瘤演变以逃避免疫监视的最重要机制之一。事实上，许多人类肿瘤中，如乳腺癌、前列腺癌、肺癌、结肠癌，以及神经母细胞瘤和黑素瘤，普遍存在IDO1的持续性高表达(Munn,D.H.；Mellor,A.L.Clin.Invest.2007,117,1147-1154.Uyttenhove,C.；Pilotte,L.；Théate,I.；Stroobant,V.；Colau,D.；Parmentier,N.；Boon,T.；Van den Eynde,J.V.Nat.Med.2003,9,1269-1274；Brody,J.R.；Costantino,C.L.；Berger,A.C.；Sato,T.；Lisanti,M.P.；Yeo,C.J.；Emmons,R.V.；Witkiewicz,A.K.Cell Cycle 2009,8,1930-1934)。IDO1是一种血红素酶，通过与分子氧结合和裂解吡咯环底物来催化L-色氨酸(L-Trp)氧化成N-甲酰犬尿氨酸(NFK)。人IDO1的晶体结构在末端血红素位点(口袋A)中具有一个结合口袋，与活性部位入口处第二口袋(口袋B)连接(
Figure PCTCN2019095516-appb-000001
U.F.；Awad,L.；Grosdidier,A.；Larrieu,P.；Stroobant,V.；Colau,D.；Cerundolo,V.；Simpson,A.J.G.；Vogel,P.；Van den Eynde,B.J.；Zoete,V.J.Med.Chem.2010,53,1172-1189)。近期研究已经证明IDO1表达由肿瘤坏死因子α(TNF-α)和其他炎症介质诱导。因此，由于初始宿主对肿瘤的炎症反应可能导致IDO1被二次诱导(Dunn,G.P.；Koebel,C.M.；Schreiber,R.D.Nat.Rev.Immunol.2006,6,836-848)。此外，增加IDO1的表达与不同肿瘤进展参数和缩短患者的存活期正相关。所有这些证据都表明抑制IDO1可能会增强癌症治疗药物的疗效。事实上，基于小鼠肿瘤模型的临床前研究表明，IDO1抑制剂与一些抗癌药物如环磷酰胺、多柔比星、紫杉醇和顺铂联合使用具有协同作用(Hou,D.Y.；Muller,A.J.；Sharma,M.D.；Du Hadaway,J.；Banerjee,T.；Johnson,M.；Mellor,A.L.；Prendergast,G.C.Cancer Res.2007,67,792-801)。鉴于IDO1酶活性介导的T细胞应答抑制以及IDO1表达与几种癌症适应症的不良预后和化疗耐药之间的相关 性，IDO1已成为癌症免疫治疗潜在的靶标。
已有小部分IDO1抑制剂进入临床研究，其中由NewLink Genetics开发的1-甲基-DL-色氨酸是第一个IDO1抑制剂(Cady,S.G.；Sono,M.Cancer Res.1991,291,326-333)。Incyte Corporation和NewLink Genetics分别开发的INCB024360和NLG919也已应用于临床试验(Mautino,M.R.；Jaipuri,F.A.；Waldo,J.；Kumar,S.；Adams,J.；van Allen,C.；Marcinowicz-Flick,A.；Munn,D.Cancer Research Philadelphia,2013,282,355-342；Nayak,A.；Hao,Z.；Sadek,R.；Vahanian,N.；Ramsey,W.；Kennedy,E.；Mautino,M.；Link,C.；Bourbo,P.；Dobbins,R.；Adams,K.；Diamond,A.；Marshall,L.；Munn,D.H.；Janik,J.Cancer 2014,2,250)。含有醌或亚胺醌基团的醌类化合物已被报道是具有高IDO1酶抑制活性的抑制剂(Bridewell,D.J.；Sperry,J.；Smith,J.R.；Kosim-Satyaputra,P.；Ching,L.-M.；Jamie,J.F.Aust.J.Chem.2013,66,40-49；
Figure PCTCN2019095516-appb-000002
E.；Larrieu,P.；Meinguet,C.；Colette,D.；Rives,A.；Blanc,S.；Ferain,T.；Pilotte,L.；Stroobant,V.；Wouters,J.Bioorg.Med.Chem.Lett.2013,23,47-54；Pasceri,R.；Siegel,D.；Ross,D.；Moody,C.J.J.Med.Chem.2013,56,3310-3317；Centko,R.M.；
Figure PCTCN2019095516-appb-000003
A.；Rosell,F.I.；Patrick,B.O.；de Voogd,N.；Mauk,A.G.Org.Lett.2014,16,6480-6483)。醌类化合物可以通过与酶活性位点的特异性相互作用或通过用还原辅因子进行氧化还原循环或通过对亲核氨基酸侧链的化学反应来抑制IDO1。
Ying Ge,Aibo Li等人在European Journal of Medicinal Chemistry,2017,128(2017)180-191中表示2-氨基酸酰胺萘醌的抗肿瘤活性有一定选择性，但其活性不高，没有明确的作用机制研究，在活性方面主要还是集中在传统的细胞毒类肿瘤药物领域。
目前临床上尚无对STAT3和IDO1双靶点抑制剂的研究，因此，研究开发具有高度专一性的STAT3和IDO1双靶点抑制剂具有重要的现实意义。
发明内容
根据本申请的一个方面，提供了具有式(I)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000004
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；和
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基。
在本申请的一个实施方案中，其中所述5元至10元杂芳基为吡啶基或噻吩基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在本申请的一个实施方案中，其中所述各个R 1均为氢。
在本申请的一个实施方案中，其中所述各个R 2均为氢。
在本申请的一个实施方案中，其中所述R 3为氢。
根据本申请的另一个方面，提供了选自以下的化合物或其药学上可接受的 盐：
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-(三氟甲基)苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3,N-二苯基-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲氧基苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲基苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氯苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-溴苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴-4-氟苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氟苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氯苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-溴苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲氧基苯 基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲氧基苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟-4-溴苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二氟苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟-4-甲基苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2,5-二甲氧基苯基)-丙酰胺；
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯-4-甲基苯基)-丙酰胺；和
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲基-4-溴苯基)-丙酰胺。
根据本申请的另一个方面，提供了具有式(II)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000005
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧 基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；和
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基。
在本申请的一个实施方案中，其中所述5元至10元杂芳基为吡啶基或噻吩基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在本申请的一个实施方案中，其中所述各个R 1均为氢。
在本申请的一个实施方案中，其中所述各个R 2均为氢。
在本申请的一个实施方案中，其中所述R 3为氢。
根据本申请的另一个方面，提供了选自以下的化合物或其药学上可接受的盐：
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-(三氟甲基)苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3,N-二苯基-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲氧基苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲基苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氯苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-溴苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴-4-氟苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氟苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氯苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-溴苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲氧基苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲氧基苯；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟-4-溴苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二氟苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟-4-甲基苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2,5-二甲氧基苯基)-丙酰胺；
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯-4-甲基苯基)-丙酰胺；和
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲基-4-溴苯基)-丙酰胺。
根据本申请的另一个方面，提供了具有式(III)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000006
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；和
R 4为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基，所述5元至10元杂芳基为吡啶基或噻吩基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在本申请的一个实施方案中，其中所述各个R 1均为氢。
在本申请的一个实施方案中，其中所述各个R 2均为氢。
在本申请的一个实施方案中，其中所述R 3为氢。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基，所述5元至10元杂芳基为噻吩基。
根据本申请的另一个方面，提供了选自以下的化合物或其药学上可接受的盐：
(R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺；
(R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺；和
(R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺。
根据本申请的另一个方面，提供了具有式(IV)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000007
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂 芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；和
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基。
在本申请的一个实施方案中，其中所述5元至10元杂芳基为吡啶基或噻吩基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在本申请的一个实施方案中，其中所述各个R 1均为氢。
在本申请的一个实施方案中，其中所述各个R 2均为氢。
在本申请的一个实施方案中，其中所述R 3为氢。
根据本申请的另一个方面，提供了(R)-N-(3,5-二甲氧基苯基)-2-((4-(甲氧基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基丙酰胺或其药学上可接受的盐。
根据本申请的另一个方面，提供了制备式(II)所述的化合物的方法，所述方法包括如下所示的步骤a)至e)：
Figure PCTCN2019095516-appb-000008
a)使邻苯二甲酸酐和L-苯丙氨酸反应，所述L-苯丙氨酸中的苯基任选地被1至n个R 1取代；
b)使步骤a)得到的产物与C 2O 2Cl 2反应；
c)使步骤b)得到的产物与X-NH 2反应；
d)使步骤c)得到的产物与肼反应；
e)使步骤d)得到的产物与1,4-萘醌反应，所述1,4-萘醌中的苯环任选地被1至m个R 2取代并且2位碳任选地被R 3取代，从而获得式(II)的化合物；
其中X、R 1、n、R 2、m和R 3为如上文中所述。
在本申请的一个实施方式中，步骤a)的反应条件为在55-85℃下、优选在70℃下在诸如乙酸或冰乙酸的酸性溶液中回流10-14小时，优选12小时；反应物的摩尔比为约1:1。
在本申请的一个实施方式中，步骤b)的反应条件为在0-15℃下、优选在0℃下在诸如二氯甲烷或三氯甲烷的卤代烷烃溶液中反应10-14小时，优选12小时；可加适量二甲基甲酰胺(DMF)作为催化剂；反应物草酰氯与另一反应物的摩尔比为2.5:1以上。
在本申请的一个实施方式中，步骤c)的反应条件为在0-15℃下、优选在0℃下在诸如二氯甲烷或三氯甲烷的卤代烷烃溶液中反应20-40分钟，优选30分钟，其中将滴速控制在每秒1滴；可加三乙胺作为缚酸剂；反应物的摩尔比为约1:1。
在本申请的一个实施方式中，步骤d)的反应条件为在室温下在诸如无水乙醇的醇类溶液中反应，反应至不再有固体析出为止，反应时间为约2.5-3.5h；反应 物水合肼与另一反应物的摩尔比为2.5:1以上。
在本申请的一个实施方式中，步骤e)的反应条件为在室温下在三乙胺、二甲基甲酰胺和水的混合溶液中反应18-24小时；反应物1,4-萘醌与另一反应物的摩尔比为约1.5:1。
根据本申请的另一个方面，提供了制备式(I)所述的化合物的方法，所述方法包括如下所示的步骤f)：
Figure PCTCN2019095516-appb-000009
所述步骤f)为将式(II)的化合物与盐酸羟胺反应，式(II)的化合物与盐酸羟胺的摩尔比为约1:3；优选地，步骤f)的反应条件为在70-80℃下在诸如无水乙醇或无水甲醇的醇类溶液中反应10-14小时，优选12小时，从而获得式(I)的化合物，其中X、R 1、n、R 2、m和R 3为如上文所述。
根据本申请的另一个方面，提供了制备式(III)所述的化合物的方法，所述方法包括如下所示的步骤g)：
Figure PCTCN2019095516-appb-000010
所述步骤g)为将式(I)的化合物与ClSO 2R 4反应，式(I)的化合物与ClSO 2R 4的摩尔比为约1:2；优选地所述步骤g)的反应条件为在诸如氩气的惰性气体中在0-15℃下、优选在0℃下在二氯甲烷中反应20-40分钟，优选30分钟，随后逐滴加入三乙胺并且反应10-20分钟；其中X、R 1、n、R 2、m、R 3和R 4为如上文中所述。
根据本申请的另一个方面，提供了制备式(IV)所述的化合物的方法，所述方法包括如下所示的步骤f’)：
Figure PCTCN2019095516-appb-000011
所述步骤f’)为将式(II)的化合物与甲氧基胺盐酸盐反应，优选地，步骤f’)的 反应条件为在80℃下在吡啶中回流反应2小时，从而获得式(IV)的化合物，其中X、R 1、n、R 2、m和R 3为如上文所述。
根据本申请的另一个方面，提供了药物组合物，其包含本申请具有活性的化合物或其药学上可接受的盐、或者根据制备式(I)和式(III)的化合物的方法得到的化合物或其药学上可接受的盐、以及一种或多种药学上可接受的载体、稀释剂、赋形剂或其组合。
根据本申请的另一个方面，提供了本申请具有活性的化合物或其药学上可接受的盐、根据制备式(I)和式(III)的化合物的方法得到的化合物或其药学上可接受的盐、或者本申请的药物组合物在抑制STAT3和/或IDO1中的用途。
根据本申请的另一个方面，提供了本申请的化合物或其药学上可接受的盐、根据制备式(I)和式(III)的化合物的方法得到的化合物或其药学上可接受的盐、或者本申请的药物组合物在制备用于抑制STAT3和/或IDO1的药物中的用途。
根据本申请的另一个方面，提供了本申请的化合物或其药学上可接受的盐、根据制备式(I)和式(III)的化合物的方法得到的化合物或其药学上可接受的盐、或者或者本申请的药物组合物在制备用于治疗癌症的药物中的用途。
在本申请的一个实施方式中，所述癌症选自结肠癌、卵巢癌、肝癌、膀胱癌、子宫颈癌和肺小细胞癌。
附图说明
图1显示用化合物Iu处理来自SKOV3细胞的全细胞裂解物的p-STAT3(Y705)、p-STAT3(S727)、总STAT3和β-肌动蛋白的蛋白质印迹。
图2显示用化合物Iu处理来自SKOV3细胞的全细胞裂解物的STAT1、STAT5和β-肌动蛋白的蛋白质印迹。
图3显示化合物Iu选择性抑制STAT3，其中使用荧光素酶法检测化合物Iu对于STAT1、STAT3、STAT4和STAT5的活性的柱状图。
图4显示化合物Iu抑制STAT3的DNA结合(ELISA测定结果)的柱状图。
图5显示化合物Iu抑制pSTAT3核转位，用IL-6刺激SKOV3细胞，用Iu(0.5μM)处理p-STAT3向细胞核的迁移。
图6显示在SKOV3癌细胞系中用Iu(0.1和0.5μM)进行细胞迁移的伤口愈合实验。
图7显示化合物Iu抑制SKOV3癌细胞系的侵袭。
图8显示化合物Iu抑制体内肝癌生长。
图9显示化合物Iu和IDO1结合的紫外吸收谱图，其表示不具有Iu化合物(蓝色)和具有2mM浓度的Iu化合物(红色)的铁IDO1的UV光谱。在存在化合物Iu的情况下，Soret峰从403nm移动到413nm。
图10显示化合物Iu和IDO1相互作用的表面等离子共振，其测定hIDO1和化合物Iu之间结合，显示了IDO1与Iu结合的SPR曲线。表明在固定有IDO-1蛋白的生物传感器芯片表面上注射的Iu浓度。测量产生的K D为0.08μM。
图11显示化合物Iu抑制体内黑色素瘤的生长。
具体实施方式
为了更好的理解本申请的实质，下面将用本申请的具体的实施方案来说明本申请的肟基萘醌衍生物、其制备方法和药理作用结果，但本申请的技术方案并不局限于此。
本申请提供了针对STAT和IDO的双靶点选择性抑制剂，尤其是针对STAT3和IDO1的双靶点选择性抑制剂。具体而言，本申请提供了肟基萘醌类化合物，其制备方法以及该类化合物和其药物组合物在制备用于治疗多种肿瘤(尤其是卵巢癌、结肠癌及肺癌等)的药物中的用途。
具体而言，本申请设计合成一系列针对STAT3和IDO1双靶点抑制剂用于肿瘤免疫治疗。通过紫外可见光光谱分析、表面等离子共振分析等作用机制研究、酶活性检测、Western Blot实验、细胞染色实验、SKOV3细胞迁移和入侵实验等细胞活性检测，发现了作用在STAT3和IDO1信号通路上的双靶点小分子抑制剂。具体而言，其可以抑制SKOV3癌细胞增殖、迁移和/或入侵。
定义
除非另外规定，本文所使用的所有技术和科学术语具有与本领域一般技术人员通常理解的相同含义。除非另外规定，所有专利、申请、公开申请和本文引用的其他公开以其整体通过引用并入本文。除非另外规定，在本文的术语有多个定义的情况下，以本部分的定义为准。
当基团被描述为“任选取代的”时，则基团可为未取代的或被一个或多个所示的取代基取代。相似地，当基团被描述为“未取代的或取代的”时，如果是取代的，则取代基可选自一种或多种所示的取代基。如果没有指出取代基，则是所述的“任选取代的”或“取代的”基团可单独和独立地被选自以下的一种或多种基团取代：烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、杂芳烷基、(杂脂环基)烷基、羟基、烃氧基、芳氧基、酰基、巯基、烷硫基、芳硫基、氰基、卤素、硫代羰基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、N-酰氨基、S-磺酰氨基、N-磺酰氨基、C-羧基、O-羧基、异氰酸基、硫氰酸基、异硫氰酸基、硝基、甲硅烷基、烃硫基、亚磺酰基、磺酰基、卤代烷基、卤代烃氧基、三卤代甲烷磺酰基、三卤代甲烷亚磺酰氨基和氨基。
“C a至C b”是指烷基、烯基或炔基中的碳原子数，或芳基、杂芳基中的碳原子数，其中“a”和“b”是整数。即，烷基、烯基、炔基、芳基、杂芳基可包含“a”至“b”个碳原子，包括“a”和“b”个碳原子。因此，例如，“C 1至C 4烷基”基团是指具有1至4个碳的所有烷基，即CH 3-、CH 3CH 2-、CH 3CH 2CH 2-、(CH 3) 2CH-、CH 3CH 2CH 2CH 2-、CH 3CH 2CH(CH 3)-和(CH 3) 3C-。如果没有指定烷基、烯基、炔基、芳基、杂芳基的“a”和“b”，则假设这些定义中所描述的是最广泛的范围。
“烷基”是指包含完全饱和(没有双键或叁键)烃基团的直链烃链或支链烃链。烷基可具有1至20个碳原子，每当在本文出现时，诸如“1至20”的数值范围是指给定范围中的各个整数；例如，“1至20个碳原子”是指可包含1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子、6个碳原子、7个碳原子、8个碳原子、9个碳原子、10个碳原子、11个碳原子、12个碳原子、13个碳原子、14个碳原子、15个碳原子、16个碳原子、17个碳原子、18个碳原子、19个碳原子或20个碳原子的烷基。烷基还可为具有1至10个碳原子的中等大小烷基。烷基还可为具有1至6个碳原子的低级烷基。化合物的烷基可以指定为“C 1-C 6烷基”或类似名称。例如，“C 1-C 4烷基”表示烷基链中有一至四个碳原子，即，烷基链选自甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基和叔丁基。典型的烷基包括但绝不限于甲基、乙基、丙基、异丙基、丁基、异丁基、叔丁基、戊基和己基。烷基可为取代的或未取代的。
“烯基”是指在直链或支链烃链中包含一个或多个双键的烃基。烯基可为未取代的或取代的。烯基可具有1至20个碳原子，每当在本文出现时，诸如“1至20”的数值范围是指给定范围中的各个整数；例如，“1至20个碳原子”是指可包含1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子、6个碳原子、7个碳原子、8个碳原子、9个碳原子、10个碳原子、11个碳原子、12个碳原子、13个碳原子、14个碳原子、15个碳原子、16个碳原子、17个碳原子、18个碳原子、19个碳原子或20个碳原子的烯基。
“炔基”是指在直链或支链烃链中包含一个或多个叁键的烃基。炔基可为未取代的或取代的。炔基可具有1至20个碳原子，每当在本文出现时，诸如“1至20”的数值范围是指给定范围中的各个整数；例如，“1至20个碳原子”是指可包含1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子、6个碳原子、7个碳原子、8个碳原子、9个碳原子、10个碳原子、11个碳原子、12个碳原子、13个碳原子、14个碳原子、15个碳原子、16个碳原子、17个碳原子、18个碳原子、19个碳原子或20个碳原子的炔基。
“芳基”是指具有遍及所有环的完全离域的π-电子体系的碳环(全部为碳)单环或多环芳香族环体系(包括其中两个碳环共享化学键的稠环体系)。芳基中的碳原子数可变化。例如，芳基可为C 6-C 14芳基、C 6-C 10芳基或C 6芳基。芳基的实例包括但不限于苯、萘和薁。芳基可为取代的或未取代的。
“杂芳基”是指包含一个或多个(例如，1个、2个、3个、4个、5个或6个)杂原 子(即除了碳之外的元素包括但不限于氮、氧和硫)的单环或多环芳香族环体系(具有完全离域的π-电子体系的环体系)。杂芳基环中的原子数可变化。例如，杂芳基在环中可包含4至14个原子、在环中可包含5至10个原子或在环中可包含5至6个原子。此外，术语“杂芳基”包括其中两个环，例如至少一个芳基环和至少一个杂芳基环或至少两个杂芳基环，共享至少一个化学键的稠环体系。杂芳基环的实例包括但不限于呋喃、呋咱、噻吩、苯并噻吩、酞嗪、吡咯、噁唑、苯并噁唑、1,2,3-噁二唑、1,2,4-噁二唑、噻唑、1,2,3-噻二唑、1,2,4-噻二唑、苯并噻唑、咪唑、苯并咪唑、吲哚、吲唑、吡唑、苯并吡唑、异噁唑、苯并异噁唑、异噻唑、三唑、苯并三唑、噻二唑、四唑、吡啶、哒嗪、嘧啶、吡嗪、嘌呤、蝶啶、喹啉、异喹啉、喹唑啉、喹喔啉、噌啉和三嗪。杂芳基可为取代的或未取代的。
“烷氧基”是指式-OR，其中R为本文定义的烷基。烷氧基的非限制性列举为甲氧基、乙氧基、正丙氧基、1-甲基乙氧基(异丙氧基)、正丁氧基、异丁氧基、仲丁氧基、叔丁氧基。烷氧基可为取代的或未取代的。
“酰基”是指通过羰基连接的作为取代基的氢、烷基、烯基、炔基或芳基。实例包括甲酰基、乙酰基、丙酰基、苯甲酰基和丙烯酰基。酰基可为取代的或未取代的。
“卤代烷基”是指其中一个或多个氢原子被卤素取代(例如，单卤代烷基、二卤代烷基和三卤代烷基)的烷基。这类基团包括但不限于氯甲基、氟甲基、二氟甲基、三氟甲基、1-氯-2-氟甲基和2-氟异丁基。卤代烷基可为取代的或未取代的。
“卤代烃氧基”是指其中一个或多个氢原子被卤素(例如，单卤代烃氧基、二卤代烃氧基和三卤代烃氧基)取代的烃氧基。这类基团包括但不限于氯甲氧基、氟甲氧基、二氟甲氧基、三氟甲氧基、1-氯-2-氟甲氧基和2-氟异丁氧基。卤代烃氧基可为取代的或未取代的。
“芳硫基”是指RS-，其中R为芳基，例如但不限于苯基。芳硫基可为取代的或未取代的。
“烃硫基”基团是指“-SR”基团，其中R可为氢、烷基(此时的“烃硫基”为“烷硫基”)、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。烃硫基可为取代的或未取代的。
“亚磺酰基”基团是指“-S(＝O)-R”基团，其中R与烃硫基中的R定义相同。亚磺酰基可为取代的或未取代的。
“磺酰基”基团是指“SO 2R”基团，其中R与烃硫基中的R的定义相同。磺酰基可为取代的或未取代的。
“O-羧基”基团是指“RC(＝O)O-”基团，其中R可为如本文定义的氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。O-羧基可为取代的或未取代的。
“硫代羰基”基团是指“-C(＝S)R”基团，其中R与关于O-羧基中的R的定义相同。 硫代羰基可为取代的或未取代的。
“三卤代甲烷磺酰基”基团是指“X 3CSO 2-“基团，其中各个X为卤素。
“三卤代甲烷亚磺酰氨基”基团是指“X 3CS(O) 2N(R A)-”基团，其中各个X为卤素，且R A为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。
术语“氨基”是指–NH 2基团。
术语“羟基”是指–OH基团。
“氰基”基团是指“-CN”基团。
“异氰酸基”基团是指“-NCO”基团。
“硫氰酸基”基团是指“-CNS”基团。
“异硫氰酸基”基团是指“-NCS”基团。
“巯基”基团是指“-SH”基团。
“羰基”基团是指C＝O基团。
“S-磺酰氨基”基团是指“-SO 2N(R AR B)”基团，其中R A和R B可独立地为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。S-磺酰氨基可为取代的或未取代的。
“N-磺酰氨基”基团是指“RSO 2N(R A)-“基团，其中R和R A可独立地为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。N-磺酰氨基可为取代的或未取代的。
“O-氨基甲酰基”基团是指“-OC(＝O)N(R AR B)”基团，其中R A和R B可独立地为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。O-氨基甲酰基可为取代的或未取代的。
“N-氨基甲酰基”基团是指“ROC(＝O)N(R A)-“基团，其中R和R A可独立地为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。N-氨基甲酰基可为取代的或未取代的。
“O-硫代氨基甲酰基”基团是指“-OC(＝S)-N(R AR B)”基团，其中R A和R B可独立地为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。O-硫代氨基甲酰基可为取代的或未取代的。
“N-硫代氨基甲酰基”基团是指“ROC(＝S)N(R A)-“基团，其中R和R A可独立地为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。N-硫代氨基甲酰基可为取代的或未取代的。
“C-酰氨基”基团是指“-C(＝O)N(R AR B)”基团，其中R A和R B可独立地为氢、烷基、烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。C-酰氨基可为取代的或未取代的。
“N-酰氨基”基团是指“RC(＝O)N(R A)-”基团，其中R和R A可独立地为氢、烷基、 烯基、炔基、环烷基、环烯基、环炔基、芳基、杂芳基、杂脂环基、芳烷基、(杂芳基)烷基或(杂脂环基)烷基。N-酰氨基可为取代的或未取代的。
如本文使用的，术语“卤素原子”或“卤素”是指元素周期表第7列的放射稳定的原子中的任一个，例如氟、氯、溴和碘。
术语“药学上可接受的盐”是指对所施用的生物体不产生显著刺激并且不破坏化合物的生物活性和性质的化合物的盐。在一些实施方案中，所述盐为化合物的酸加成盐。能通过使化合物与无机酸反应获得药物盐，所述无机酸例如氢卤酸(例如，氢氯酸或氢溴酸)、硫酸、硝酸和磷酸。能通过使化合物与有机酸反应获得药物盐，所述有机酸例如脂肪族或芳香族的羧酸或磺酸，例如甲酸、乙酸、琥珀酸、乳酸、苹果酸、酒石酸、柠檬酸、抗坏血酸、烟酸、甲烷磺酸、乙烷磺酸、对甲苯磺酸、水杨酸或萘磺酸。还能通过使化合物与碱反应以形成盐来获得药物盐，例如铵盐、诸如钠盐或钾盐的碱金属盐、诸如钙盐或镁盐的碱土金属盐、诸如二环己基胺、N-甲基-D-葡糖胺、三(羟基甲基)甲基胺、C 1-C 7烷基胺、环己基胺、三乙醇胺、乙二胺的有机碱盐和与诸如精氨酸和赖氨酸的氨基酸的盐。
除非另外规定，本申请中使用的术语和短语及其变型，特别是在所附的权利要求中应被解释为开放式的而非限定。作为前述的实例，术语“包括”应被理解为是指“包括而不限于”同义并且为包括式或开放式，并且不排除附加、未列举的元素或方法步骤；术语“具有”应被解释“具有至少”。
应当理解，在本文描述的具有一个或多个手性中心的任何化合物中，如果未清楚表明绝对立体化学构型，则各个中心可独立地为R-构型或S-构型或其混合物。因此，本文提供的化合物可为对映异构体纯的、对映异构体富集的、外消旋混合物、非对映异构体纯的、非对映异构体富集的或立体异构体混合物。此外，应当理解，在本文描述的产生可被定义为E或Z的几何异构体的具有一个或多个双键的任何化合物中，各个双键可独立地为E或Z、其混合物。
化合物
本文公开的一些实施方案涉及具有式(I)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000012
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；和
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基。
在一些实施方案中，其中所述C 6-C 10芳基为苯基。
在一些实施方案中，其中5元至10元杂芳基为吡啶基或噻吩基。
在一些实施方案中，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在一些实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在一些实施方案中，其中所述各个R 1均为氢。
在一些实施方案中，其中所述各个R 2均为氢。
在一些实施方案中，其中所述R 3为氢。
本文公开的一些实施方案涉及具有式(II)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000013
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；和
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基。
在一些实施方案中，其中所述C 6-C 10芳基为苯基。
在一些实施方案中，其中5元至10元杂芳基为吡啶基或噻吩基。
在一些实施方案中，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在一些实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在一些实施方案中，其中所述各个R 1均为氢。
在一些实施方案中，其中所述各个R 2均为氢。
在一些实施方案中，其中所述R 3为氢。
本文公开的一些实施方案涉及具有式(III)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000014
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代 的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；和
R 4为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在一些实施方案中，其中所述C 6-C 10芳基为苯基，5元至10元杂芳基为吡啶基或噻吩基。
在一些实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在一些实施方案中，其中所述各个R 1均为氢。
在一些实施方案中，其中所述各个R 2均为氢。
在一些实施方案中，其中所述R 3为氢。
在一些实施方案中，其中所述C 6-C 10芳基为苯基，所述5元至10元杂芳基为噻吩基。
本文公开的一些实施方案涉及具有式(IV)的化合物或其药学上可接受的盐：
Figure PCTCN2019095516-appb-000015
其中
X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
n为选自1、2、3、4或5的整数；
各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
m为选自1、2、3或4的整数；和
R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基。
在本申请的一个实施方案中，其中所述5元至10元杂芳基为吡啶基或噻吩基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任 选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
在本申请的一个实施方案中，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
在本申请的一个实施方案中，其中所述各个R 1均为氢。
在本申请的一个实施方案中，其中所述各个R 2均为氢。
在本申请的一个实施方案中，其中所述R 3为氢。在上述的实施方案中，X任选地被1个、2个、3个、4个、5个、6个、7个、8个、9个、或10个基团取代。
合成
本申请的具体实验方法以邻苯二甲酸酐和L-苯丙氨酸为基础，合成式(I)、式(II)、式(III)和(IV)所示的肟基萘醌衍生物。
本申请的式(I)、式(II)、式(III)和式(IV)所示的肟基萘醌衍生物的合成路线如下(其中X、R 1、n、R 2、m、R 3和R 4均如上文所述)：
Figure PCTCN2019095516-appb-000016
其中
针对式(II)的化合物，2-氨基酸酰胺-萘醌类化合物(本申请的化合物IIa-IIz)，其经过步骤a)至e)制备：
a)反应物为邻苯二甲酸酐和L-苯丙氨酸(L-苯丙氨酸中的苯基任选地被1至n个R 1取代)，反应条件：70℃，乙酸中回流12h；
b)与C 2O 2Cl 2反应，反应条件：DCM(二氯甲烷)，DMF(二甲基甲酰胺)，0℃，12h；
c)与X-NH 2(X为任选取代的C 6-C 10芳基或5元至10元杂芳基，例如苯胺)反应，反应条件：三乙胺，DCM，0℃；将滴速控制在每秒1滴，反应时间为约30min。
d)与肼(NH 2NH 2)反应，反应条件：无水乙醇，室温；反应时间为约2.5～3.5h。
e)与1,4-萘醌(1,4-萘醌中的苯环任选地被1至m个R 2取代，2位碳被R 3取代)反应，反应条件：DMF，水，三乙胺，室温，18-20h。
针对式(I)的化合物，2-氨基酸酰胺-肟基萘醌类化合物(化合物Ia-Iz)，其经过步骤f)制备：上述步骤e)的产物与盐酸羟胺(H 3NO·HCl)反应，反应条件为在70℃-80℃下，在无水乙醇中回流12h。
针对式(III)的化合物，2-氨基酸酰胺-羟肟酯基萘醌类化合物(化合物IIIk-1、IIIq、IIIr、IIIk-2)，其经过步骤g)制备：上述步骤f)的产物与ClSO 2R 4(R 4为任选地取代的C6-C10芳基或5元至10元杂芳基，例如4-叔丁基苯磺酰氯)反应，反应条件：DCM，氩气，三乙胺，水，0℃，反应时间：约40min。
针对式(III)的化合物，其经过步骤f’)制备：上述步骤e)的产物与甲氧胺盐酸盐反应，反应条件为在80℃下，在吡啶中回流12h。
以上反应中各反应物的R 1-R 4、X、n和m如上文所述。
组合物
本申请具有活性的化合物用作药物时，可以直接使用，或者以药物组合物的形式使用。该药物组合物含有0.1-99％，优选为0.5-90％的本申请具有活性的化合物，其余为药物学上可接受的、对人和动物无毒和惰性的可药用的载体。
任选地，该药物组合物中含有的活性化合物的比例为1％、10％、20％、30％、40％、50％、60％、70％、80％、90％、95％、或99％。
药学上可接受的载体是一种或多种固体、半固体和液体稀释剂、填料以及药物制品辅料。本申请的药物组合物以单位体重服用量的形式使用。应用本申请的药物可采用本领域惯用的剂型，例如：药膏、片剂、丸剂、栓剂乳剂、输入液和注射液等。这些剂型按照众所周知的方法，使用传统的添加剂和赋型剂制得。由此制得的药物根据需要可按局部、非肠道、口服等途径给药。
本申请具有活性的化合物的施用量可根据用药途径、患者的年龄、体重、所治疗的疾病的类型和严重程度等变化，其日剂量可以是0.01-10mg/kg体重，优选0.1-5mg/kg体重。可以一次或多次施用。
实施例
本申请实施例中所用试剂的来源和规格如下。
1,4-萘醌(规格：25g)、盐酸羟胺(规格：25g)、4-叔丁基苯磺酰氯(规格：25g)、 L-苯丙氨酸(规格：500g)、甲氧基胺盐酸盐(规格：25g)、邻苯二甲酸酐(规格：500g)、草酰氯(规格：500ml)购自上海阿拉丁生化科技股份有限公司；
三乙胺(规格：500ml)、二氯甲烷(规格：500ml)、N,N-二甲基甲酰胺(规格：500ml)、冰醋酸(规格：500ml)、盐酸(规格：500ml)、无水硫酸钠(规格：500g)、无水乙醇(规格：500ml)、乙酸乙酯(规格：5L)、石油醚(规格：5L)、吡啶(规格：500ml)购自广东汕头西陇科学股份有限公司；
各种芳香胺(规格：25g或25ml)、异辛胺(规格：25g)，正丙胺(规格：250ml)购自成都安耐吉试剂有限公司采购；
SHB-III循环水多用真空泵、旋转蒸发器(郑州长城科工贸有限公司)；电子智能控温仪、85-1A型磁力搅拌器(巩义市予华仪器有限责任公司)；AVANCE AV400超导核磁共振仪(瑞士，布鲁克公司)；EL104电子天平(梅特勒—托利多仪器有限公司)；KQ5200E型超声波清洗器(昆山市超声仪器有限公司)；DHG-9146A型电热恒温鼓风干燥箱(上海精宏设备有限公司)；RY-1熔点仪(天津市天分分析仪器厂，温度计未校正)；SHIMADZU QP5050A型质谱仪(APCI)；微量旋光仪(大昌华嘉商业有限公司)；紫外仪(Agilent Technologies Cary60 UV-Vis)。
水合肼(规格：500ml)购自国药集团化学试剂有限公司；
以上试剂均为分析纯。
实施例1式(II)的化合物(化合物IIa-IIz)(2-氨基酸酰胺-萘醌类化合物)的合成
取一个250mL的圆底烧瓶，向其中倒入80mL的冰醋酸，再加入10g L-苯丙氨酸和8.966g邻苯二甲酸酐，在70℃油浴锅中回流搅拌12h。待反应结束后，使其冷却至常温，然后向圆底烧瓶中缓慢加入150mL蒸馏水，并搅拌直至白色晶体析出完全，减压抽滤即可得到白色针状的2-(1,3-二氧代异二氢吲哚-2-基)-3-苯基丙酸晶体，在55℃下烘干。
取上述产物2-(1,3-二氧代异二氢吲哚-2-)-3-苯基丙酸1g加入装有25mL二氯甲烷的100mL圆底烧瓶中，冰浴条件下用磁力搅拌器下搅拌，待圆底烧瓶中的固体全部溶解后，向其中加入5mL草酰氯，再加入1滴DMF做催化剂，套上干燥管，使其反应12个小时，得到氯化产物。
将反应混合液减压旋转干燥，用二氯甲烷溶解，转移至恒压滴定管，并在冰浴条件下滴加入装有1mL三乙胺和NH 2-X(反应物按1:1的量)加入圆底烧瓶中，控制每秒一滴的滴速，滴加完后继续搅拌，待气体挥发完后，减压旋转干燥。
NH 2-X例如为4-(三氟甲基)苯胺、2-氟苯胺、苯胺、3-氟苯胺、2-甲氧基苯胺、3,5-二甲基苯胺、2-甲苯胺、4-氯苯胺、4-溴苯胺、3-溴-4-氟苯胺、3-(三氟甲基)苯胺、4-氟苯胺、3-甲苯胺、2-氯苯胺、3-氯苯胺、2-溴苯胺、3-溴苯胺、3-甲氧基苯胺、4-甲氧基苯胺、4-甲苯胺、3,5-二甲氧基苯胺、3-氟-4-溴苯胺、3,5-二氟 苯胺、2-氟-4-甲基苯胺、2,5-二甲氧基苯胺、3-氯-4-甲基苯胺、2-甲基-4-溴苯胺。
用25mL无水乙醇溶解，常温搅拌下加入4mL水合肼，反应至不再有固体析出为止，获得粗产物。减压旋转干燥，用水和二氯甲烷萃取三次(3×30mL)，收集下层的有机液，弃去水层，有机层用无水硫酸钠干燥2h，经洗脱剂为石油醚:乙酸乙酯(4:1)的硅胶柱纯化，得到如下纯化产物：(R)-2-氨基-3-苯基-N-(4-(三氟甲基)苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(2-氟苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-苯基-丙酰胺、(R)-2-氨基-3-苯基-N-(3-氟苯基)苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(2-甲氧基苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3,5-二甲基苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(2-甲苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(4-氯苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(4-溴苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-溴-4-氟苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(4-氟苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-甲苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(2-氯苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-氯苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(2-溴苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-溴苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-甲氧基苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(4-甲氧基苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(4-甲苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3,5-二甲氧基苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-氟-4-溴苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3,5-二氟苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(2-氟-4-甲基苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(2,5-二甲氧基苯基)-丙酰胺、(R)-2-氨基-3-苯基-N-(3-氯-4-甲基苯基)-丙酰胺、和(R)-2-氨基-3-苯基-N-(2-甲基-4-溴苯基)-丙酰胺。
将上述产物与1,4-萘醌以1:1.5的摩尔比加入装有0.5mL三乙胺、15mL DMF和8-10mL蒸馏水的圆底烧瓶，在常温下搅拌反应18-20小时，TLC检测反应进程。反应完成后用1mol/L盐酸调节其pH为3-4，用水和二氯甲烷萃取三次(3×30mL)，用无水硫酸钠干燥2h，减压旋转干燥，经洗脱剂为石油醚:乙酸乙酯(4:1)的硅胶柱纯化，旋转干燥得黄色固体。所得产物具有如下具体结构式和参数表征。
式(II)的化合物的具体结构式及参数表征如下：
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-(三氟甲基)苯基)-丙酰胺(IIa)
Figure PCTCN2019095516-appb-000017
产率：45.8％。黄色固体。熔点210.2～211.0℃。0 1H NMR(400MHz,DMSO):δ10.50(s,1H),7.98(d,J＝6.9Hz,1H),7.92–7.86(m,1H),7.80(ddd,J＝8.6,6.7,2.8Hz,3H),7.74–7.67(m,3H),7.35–7.24(m,5H),7.18(t,J＝7.1Hz,1H),5.70(s,1H),4.49(q,J＝7.0Hz,1H),3.30(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.84,180.98,169.27,147.57,141.82,136.83,134.92,132.65,132.50,130.19,129.23,129.23,128.30,128.30,126.72,126.16,126.12,126.00,125.37,123.75,122.93,119.61,119.61,101.06,57.65,37.01。HR-MS(m/z)(ESI)：计算为C 26H 19F 3N 2O 3Na[M+Na] +:487.1240；实测值：487.1223。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟苯基)-丙酰胺(IIb)
Figure PCTCN2019095516-appb-000018
产率：70.2％。黄色固体。熔点179.2～179.9℃。 1H NMR(400MHz,DMSO):δ10.06(s,1H),7.97(d,J＝7.4Hz,1H),7.90(d,J＝6.9Hz,1H),7.83–7.76(m,2H),7.72(td,J＝7.5,1.1Hz,1H),7.34(d,J＝7.2Hz,2H),7.27(dd,J＝13.5,6.0Hz,4H),7.22–7.15(m,3H),5.73(s,1H),4.63(dd,J＝14.0,8.0Hz,1H),3.28(t,J＝6.2Hz,2H)。 13C NMR(101MHz,DMSO):δ181.86,181.03,169.21,155.17,152.73,147.59,136.93,134.96,132.67,132.52,130.21,129.30,129.30,128.38,128.28,126.69,126.01,125.39,124.53,124.47,115.75,115.56,101.09,57.17,37.18。HR-MS(m/z)(ESI)：计算为C 25H 20FN 2O 3[M+H] +:415.1452；实测值： 415.1437。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3,N-二苯基-丙酰胺(IIc)
Figure PCTCN2019095516-appb-000019
产率：70.2％。黄色固体。熔点99.7～100.2℃。 1H NMR(400MHz,DMSO):δ10.17(s,1H),8.00–7.95(m,1H),7.90(dd,J＝7.6,0.7Hz,1H),7.79(td,J＝7.5,1.2Hz,1H),7.71(td,J＝7.5,1.2Hz,1H),7.55(d,J＝7.7Hz,2H),7.36–7.29(m,4H),7.26(t,J＝7.4Hz,2H),7.19(d,J＝7.6Hz,2H),7.08(t,J＝7.4Hz,1H),5.70(s,1H),4.45(q,J＝7.0Hz,1H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.82,181.00,168.55,147.50,138.24,136.92,134.91,132.68,132.46,130.19,130.19,129.24,128.80,128.80,128.28,128.28,126.68,125.98,125.37,123.91,119.74,119.74,100.99,57.55,37.19。HR-MS(m/z)(ESI)：计算为C 25H 21N 2O 3[M+H] +:397.1547；实测值：397.1531。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟苯基)-丙酰胺(IId)
Figure PCTCN2019095516-appb-000020
产率：61.4％。黄色固体。熔点123.5～124.5℃。 1H NMR(400MHz,DMSO):δ10.42(s,1H),7.97(d,J＝7.5Hz,1H),7.91–7.85(m,1H),7.85–7.77(m,1H),7.72(td,J＝7.5,1.2Hz,1H),7.54(dd,J＝11.5,1.9Hz,1H),7.38–7.24(m,7H),7.18(t,J＝7.1Hz,1H),6.92(td,J＝8.4,2.3Hz,1H),5.67(s,1H),4.44(dd,J＝14.7, 7.0Hz,1H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.99,181.08,169.10,163.37,160.97,147.70,140.09,139.99,136.97,135.08,132.72,132.66,130.70,130.28,129.34,128.42,126.83,126.13,125.49,115.51,110.63,106.67,101.08,57.69,37.10。HR-MS(m/z)(ESI)：计算为C 25H 20FN 2O 3[M+H] +:415.1452；实测值：415.1437。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲氧基苯基)-丙酰胺(IIe)
Figure PCTCN2019095516-appb-000021
产率：72.9％。黄色固体。熔点133.6～134.6℃。 1H NMR(400MHz,DMSO):δ9.54(s,1H),7.96(d,J＝7.0Hz,1H),7.93–7.82(m,2H),7.79(td,J＝7.5,1.0Hz,1H),7.70(td,J＝7.5,1.1Hz,1H),7.34(d,J＝7.3Hz,2H),7.30–7.17(m,4H),7.09(dd,J＝11.3,4.2Hz,1H),7.03(d,J＝7.2Hz,1H),6.96–6.88(m,1H),5.79(s,1H),4.73(dd,J＝14.0,8.0Hz,1H),3.87–3.67(m,3H),3.25(t,J＝6.1Hz,2H)。 13C NMR(101MHz,DMSO):δ181.84,181.07,168.87,150.05,147.47,137.02,134.91,132.68,132.44,130.16,129.31,129.31,128.22,128.22,126.60,126.45,125.96,125.36,125.13,122.37,120.27,111.36,101.26,57.16,55.77,37.31。HR-MS(m/z)(ESI)：计算为C 26H 23N 2O 4[M+H] +:427.1652；实测值：427.1638。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲基苯基)-丙酰胺(IIf)
Figure PCTCN2019095516-appb-000022
产率：63.7％。黄色固体。熔点119.9～120.8℃。 1H NMR(400MHz,DMSO):δ10.03(s,1H),7.96(dd,J＝7.6,0.9Hz,1H),7.89(dd,J＝7.6,1.0Hz,1H),7.79(td,J＝7.5,1.2Hz,1H),7.70(td,J＝7.5,1.3Hz,1H),7.34–7.30(m,2H),7.27(t,J＝7.5Hz,2H),7.20(q,J＝7.7Hz,4H),6.71(s,1H),5.68(s,1H),4.42(dd,J＝14.2,7.6Hz,1H),3.30–3.17(m,2H),2.22(s,6H)。 13C NMR(101MHz,DMSO):δ181.80,180.99,168.43,147.53,138.14,137.81,137.81,137.01,134.90,132.67,132.45,130.18,130.18,129.24,129.24,128.29,128.29,126.66,125.97,125.37,125.37,117.40,100.97,57.66,37.17,21.02,21.02。HR-MS(m/z)(ESI)：计算为C 27H 25N 2O 3[M+H] +:425.1860；实测值：425.1843。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氯苯基)-丙酰胺(IIh)
Figure PCTCN2019095516-appb-000023
产率：71.3％。黄色固体。熔点194.3～194.1℃。 1H NMR(400MHz,DMSO):δ10.29(s,1H),7.96(d,J＝7.5Hz,1H),7.89(d,J＝6.9Hz,1H),7.79(dd,J＝10.8,4.1Hz,1H),7.71(t,J＝7.5Hz,1H),7.59(d,J＝8.9Hz,2H),7.37(d,J＝8.8Hz,2H),7.31(d,J＝7.0Hz,2H),7.25(dd,J＝12.7,5.0Hz,3H),7.18(t,J＝7.1Hz,1H),5.69(s,1H),4.44(q,J＝7.0Hz,1H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.84,180.98,168.76,147.55,137.20,136.91,134.91,132.66,132.48,130.19,129.24,129.24,128.74,128.74,128.30,128.30,127.54,126.71,125.99,125.38,121.28,121.28,101.01,57.59,37.09。HR-MS(m/z)(ESI)：计算为C 25H 19ClN 2O 3Na[M+Na] +:453.0976；实测值：453.0958。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-溴苯基)-丙酰胺(IIi)
Figure PCTCN2019095516-appb-000024
产率：68.5％。黄色固体。熔点207.2～208.1℃。 1H NMR(400MHz,DMSO):δ10.27(s,1H),7.98(dd,J＝7.6,1.0Hz,1H),7.89(dt,J＝8.7,4.4Hz,1H),7.81(td,J＝7.5,1.3Hz,1H),7.73(td,J＝7.5,1.3Hz,1H),7.52(d,J＝2.1Hz,3H),7.30(dd,J＝10.4,3.6Hz,2H),7.27–7.16(m,3H),5.68(s,1H),4.44(q,J＝7.0Hz,1H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.82,180.99,168.75,147.57,137.60,136.89,134.94,132.66,132.51,131.65,131.65,130.20,129.23,129.23,128.29,128.29,126.69,126.01,125.38,121.64,121.64,115.56,100.98,57.59,37.05。HR-MS(m/z)(ESI)：计算为C 25H 20BrN 2O 3[M+H] +:475.0652；实测值：475.0628。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴-4-氟苯基)-丙酰胺(IIj)
Figure PCTCN2019095516-appb-000025
产率：69.9％。黄色固体。熔点244.2～245.2℃。1H NMR(400MHz,DMSO):δ10.45(s,1H),7.96(dd,J＝7.6,1.0Hz,1H),7.89(dd,J＝7.6,1.0Hz,1H),7.79(td,J＝7.5,1.3Hz,1H),7.71(ddd,J＝9.4,5.8,1.7Hz,2H),7.64(t,J＝8.4Hz,1H),7.29(ddd,J＝18.4,11.7,4.6Hz,6H),7.21–7.15(m,1H),5.68(s,1H),4.45(q,J＝7.0Hz,1H),3.28(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.83,180.94,169.15,159.21,156.79,147.57,139.45,136.84,134.90,133.43,132.62,132.48,130.17,129.21,129.21,128.29,128.29,126.71,125.98,125.36,116.99,116.96, 107.79,101.05,57.63,36.95。HR-MS(m/z)(ESI)：计算为C 25H 17BrFN 2O 3[M-H] +:491.0412；实测值：491.0414。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)丙酰胺(IIk)
Figure PCTCN2019095516-appb-000026
产率：46.5％。黄色固体。熔点160.8～161.5℃。 1H NMR(400MHz,DMSO):δ10.47(s,1H),8.02–7.96(m,2H),7.89(dd,J＝7.6,1.0Hz,1H),7.83–7.76(m,2H),7.71(dd,J＝7.5,1.3Hz,1H),7.57(t,J＝8.0Hz,1H),7.44(d,J＝7.8Hz,1H),7.34–7.29(m,2H),7.26(dd,J＝8.1,6.7Hz,3H),7.19(ddd,J＝7.1,3.8,1.3Hz,1H),5.69(s,1H),4.47(dd,J＝14.8,7.1Hz,1H),3.30(d,J＝6.9Hz,2H)。 13C NMR(101MHz,DMSO):δ181.88,180.99,169.21,147.63,139.03,136.91,134.94,132.67,132.53,130.21,130.14,129.24,129.24,128.32,128.32,126.73,126.01,125.39,123.29,120.28,120.25,115.81,115.77,101.07,57.66,37.01。HR-MS(m/z)(ESI)：计算为C 26H 19F 3N 2O 3Na[M+Na] +:465.1421；实测值：465.1401。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氟苯基)-丙酰胺(IIl)
Figure PCTCN2019095516-appb-000027
产率：76.6％。黄色固体。熔点193.2～194.2℃。 1H NMR(400MHz,DMSO):δ10.22(s,1H),7.97(dd,J＝7.6,1.1Hz,1H),7.89(dd,J＝7.6,1.1Hz,1H),7.80(td, J＝7.5,1.3Hz,1H),7.74–7.69(m,1H),7.59–7.53(m,2H),7.33–7.22(m,5H),7.21–7.13(m,3H),5.69(s,1H),4.43(dd,J＝14.8,7.0Hz,1H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.83,181.00,168.50,159.57,157.18,147.56,136.96,134.93,132.68,132.49,130.20,129.25,129.25,128.30,128.30,126.69,126.00,125.38,121.64,121.56,115.53,115.31,100.98,57.53,37.13。HR-MS(m/z)(ESI)：计算为C 25H 20FN 2O 3[M+H] +:415.1452；实测值：415.1436。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲苯基)-丙酰胺(IIm)
Figure PCTCN2019095516-appb-000028
产率：60.2％。黄色固体。熔点145.3～145.0℃。 1H NMR(400MHz,DMSO):δ10.12(s,1H),7.98–7.92(m,1H),7.88(d,J＝7.0Hz,1H),7.76(td,J＝7.5,1.1Hz,1H),7.67(td,J＝7.5,1.1Hz,1H),7.43–7.36(m,2H),7.33(d,J＝7.2Hz,2H),7.30–7.16(m,5H),6.89(d,J＝7.5Hz,1H),5.70(s,1H),4.45(dd,J＝14.4,7.3Hz,1H),3.28(d,J＝6.0Hz,2H),2.26(s,3H)。 13C NMR(101MHz,DMSO):δ181.82,180.99,168.53,147.52,138.22,138.06,137.00,134.87,132.68,132.42,130.17,129.27,129.27,128.66,128.30,128.30,126.69,125.96,125.37,124.59,120.22,116.88,101.01,57.65,37.21,21.12。HR-MS(m/z)(ESI)：计算为C 26H 23N 2O 3[M+H] +:411.1703；实测值：411.1688。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氯苯基)-丙酰胺(IIn)
Figure PCTCN2019095516-appb-000029
产率：71.8％。黄色固体。熔点185.8～186.6℃。 1H NMR(400MHz,DMSO):δ9.88(s,1H),8.00–7.95(m,1H),7.91(dd,J＝7.6,1.0Hz,1H),7.80(td,J＝7.5,1.3Hz,1H),7.72(td,J＝7.5,1.3Hz,1H),7.59(dd,J＝8.0,1.4Hz,1H),7.49(dd,J＝8.0,1.4Hz,1H),7.36(t,J＝5.6Hz,2H),7.33–7.26(m,4H),7.25–7.18(m,2H),5.81(s,1H),4.64(dd,J＝14.1,7.9Hz,1H),3.34–3.26(m,2H)。 13C NMR(101MHz,DMSO):δ181.86,181.03,169.19,147.57,136.94,134.92,134.11,132.66,132.50,130.20,129.55,129.32,129.32,128.32,128.32,127.50,127.11,126.96,126.71,126.53,125.98,125.38,101.41,57.25,37.14。HR-MS(m/z)(ESI)：计算为C 25H 20ClN 2O 3[M+H] +:431.1157；实测值：431.1142。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯苯基)-丙酰胺(IIo)
Figure PCTCN2019095516-appb-000030
产率：63.3％。黄色固体。熔点191.6～192.2℃。 1H NMR(400MHz,DMSO):δ10.33(s,1H),7.97(dd,J＝7.6,1.0Hz,1H),7.89(dd,J＝7.6,1.0Hz,1H),7.80(td,J＝7.5,1.3Hz,1H),7.75(t,J＝2.0Hz,1H),7.72(td,J＝7.5,1.4Hz,1H),7.44(ddd,J＝8.2,1.9,1.0Hz,1H),7.38–7.31(m,2H),7.31–7.24(m,4H),7.21–7.12(m,2H),5.69(s,1H),4.44(q,J＝7.0Hz,1H),3.28(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.85,180.97,168.99,147.58,139.68,136.88,134.92,133.11, 132.65,132.50,130.54,130.20,129.23,129.23,128.30,128.30,126.71,126.00,125.38,123.62,119.17,118.07,101.04,57.63,37.02。HR-MS(m/z)(ESI)：计算为C 25H 20ClN 2O 3[M+H] +:431.1157；实测值：431.1143。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-溴苯基)-丙酰胺(IIp)
Figure PCTCN2019095516-appb-000031
产率：75.3％。黄色固体。熔点178.5～178.6℃。 1H NMR(400MHz,DMSO):δ9.85(s,1H),7.98(dd,J＝7.6,1.1Hz,1H),7.91(dd,J＝7.6,1.1Hz,1H),7.82(td,J＝7.5,1.3Hz,1H),7.76–7.71(m,1H),7.66(dd,J＝8.0,1.3Hz,1H),7.51(dd,J＝8.0,1.5Hz,1H),7.40–7.35(m,3H),7.32–7.25(m,3H),7.23–7.19(m,1H),7.20–7.14(m,1H),5.81(s,1H),4.60(dd,J＝14.3,7.7Hz,1H),3.31(s,1H)。 13C NMR(101MHz,DMSO):δ181.85,181.02,169.08,147.58,136.99,135.45,134.94,132.73,132.66,132.52,130.20,129.30,129.30,128.33,128.33,128.11,127.58,127.23,126.70,125.99,125.38,118.15,101.47,57.28,37.05。HR-MS(m/z)(ESI)：计算为C 25H 20BrN 2O 3[M+H] +:475.0652；实测值：475.0632。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺(IIq)
Figure PCTCN2019095516-appb-000032
产率：66.9％。黄色固体。熔点184.0～184.7℃。 1H NMR(400MHz,DMSO): δ10.31(s,1H),7.98(dd,J＝7.6,1.0Hz,1H),7.90(dd,J＝8.8,1.3Hz,2H),7.81(td,J＝7.5,1.4Hz,1H),7.72(td,J＝7.5,1.4Hz,1H),7.49(dt,J＝7.1,2.2Hz,1H),7.32–7.30(m,2H),7.29–7.24(m,4H),7.19(ddd,J＝7.1,3.9,1.4Hz,1H),5.68(s,1H),4.44(q,J＝7.0Hz,1H),3.28(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.84,180.97,168.95,147.58,139.81,136.88,134.93,132.65,132.51,130.84,130.19,129.22,129.22,128.30,128.30,126.70,126.51,126.00,125.38,122.02,121.53,118.45,101.03,57.62,37.00。HR-MS(m/z)(ESI)：计算为C 25H 20BrN 2O 3[M+H] +:475.0652；实测值：475.0632。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺(IIr)
Figure PCTCN2019095516-appb-000033
产率：68.9％。黄色固体。熔点111.9～112.5℃。 1H NMR(400MHz,DMSO):δ10.17(s,1H),7.96(dd,J＝7.7,1.1Hz,1H),7.89(dd,J＝7.6,1.0Hz,1H),7.78(td,J＝7.5,1.3Hz,1H),7.70(td,J＝7.5,1.4Hz,1H),7.34–7.30(m,2H),7.29–7.17(m,6H),7.14–7.10(m,1H),6.66(ddd,J＝8.2,2.5,0.7Hz,1H),5.69(s,1H),4.44(dd,J＝14.8,7.1Hz,1H),3.38(s,3H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.81,180.98,168.61,159.52,147.50,139.44,136.95,134.89,132.66,132.44,130.17,129.63,129.24,128.28,126.67,125.97,125.36,111.90,109.36,105.43,100.98,57.61,55.00,37.16。 13C NMR(101MHz,DMSO):δ181.81,180.98,168.61,159.52,147.50,139.44,136.95,134.89,132.66,132.44,130.17,129.63,129.24,129.24,128.28,128.28,126.67,125.97,125.36,111.90,109.36,105.43,100.98,57.61,55.00,37.16。HR-MS(m/z)(ESI)：计算为C 26H 23N 2O 4[M+H] +:427.1652；实测值：427.1637。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲氧基苯基)-丙酰胺(IIs)
Figure PCTCN2019095516-appb-000034
产率：78.9％。黄色固体。熔点148.3～148.7℃。 1H NMR(400MHz,DMSO):δ10.06(s,1H),7.94(dd,J＝7.6,1.0Hz,1H),7.88(dd,J＝7.6,0.9Hz,1H),7.76(td,J＝7.5,1.3Hz,1H),7.68(td,J＝7.5,1.3Hz,1H),7.47–7.42(m,2H),7.30(dd,J＝12.3,5.4Hz,2H),7.26(t,J＝7.4Hz,2H),7.21–7.15(m,2H),6.92–6.86(m,2H),5.70(s,1H),4.42(q,J＝7.0Hz,1H),3.70(d,J＝3.7Hz,4H),3.26(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ181.88,181.03,168.11,155.77,147.51,137.00,134.92,132.73,132.47,131.33,130.21,129.31,129.31,128.33,128.33,126.73,126.01,125.41,121.44,121.44,113.98,113.98,101.00,57.50,55.21,37.31。HR-MS(m/z)(ESI)：计算为C 26H 23N 2O 4[M+H] +:427.1652；实测值：427.1637。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲苯基)-丙酰胺(IIt)
Figure PCTCN2019095516-appb-000035
产率：70.5％。黄色固体。熔点97.2～98.1℃。 1H NMR(400MHz,DMSO):δ10.10(s,1H),7.94(dd,J＝7.6,0.9Hz,1H),7.88(dd,J＝7.6,0.8Hz,1H),7.76(td,J＝7.5,1.3Hz,1H),7.67(td,J＝7.5,1.3Hz,1H),7.45(d,J＝8.4Hz,2H),7.32(d,J＝7.1Hz,2H),7.26(t,J＝7.5Hz,2H),7.18(dt,J＝9.6,6.5Hz,2H),7.11(d,J＝8.3Hz,2H),5.70(s,1H),4.44(dd,J＝14.7,7.1Hz,1H),3.27(d,J＝6.8Hz,2H),2.23(s,3H)。 13C NMR(101MHz,DMSO):δ181.79,180.96,168.31,147.46,136.97,135.73,134.84,132.90,132.67,132.39,130.16,129.25,129.25,129.16,129.16,128.27,128.27,126.66,125.94,125.34,119.75,119.75,100.97,57.54,37.25,20.44。HR-MS(m/z)(ESI)：计算为C 26H 22N 2O 3Na[M+Na] +:433.1523；实测值： 433.1505。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲氧基苯基)-丙酰胺(IIu)
Figure PCTCN2019095516-appb-000036
产率：73.0％。黄色固体。熔点195.3～195.5℃。 1H NMR(400MHz,DMSO):δ10.04(s,1H),7.97(dd,J＝7.6,0.9Hz,1H),7.90(dd,J＝7.6,1.0Hz,1H),7.81(td,J＝7.5,1.3Hz,1H),7.72(td,J＝7.5,1.3Hz,1H),7.29(dt,J＝14.9,4.6Hz,4H),7.23–7.16(m,3H),7.08(dd,J＝8.7,2.3Hz,1H),6.90(d,J＝8.8Hz,1H),5.68(s,1H),4.41(dd,J＝14.7,7.0Hz,1H),3.72(d,J＝4.7Hz,6H),3.26(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ179.89,169.77,160.55,160.55,145.26,140.46,140.12,137.17,133.88,132.95,129.23,129.23,129.01,128.45,128.35,128.35,126.68,125.86,122.35,97.89,97.89,95.82,91.41,57.89,55.16,55.16,37.62。HR-MS(m/z)(ESI)：计算为C 27H 25N 2O 5[M+H] +:457.1758；实测值：457.1740。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟-4-溴苯基)-丙酰胺(IIv)
Figure PCTCN2019095516-appb-000037
产率：55.3％。黄色固体。熔点226.7～227.5℃。 1H NMR(400MHz,DMSO):δ10.47(s,1H),7.96(dd,J＝7.6,1.0Hz,1H),7.89(dd,J＝7.6,1.0Hz,1H),7.79(td, J＝7.5,1.3Hz,1H),7.71(ddd,J＝8.6,4.9,1.9Hz,2H),7.63(t,J＝8.4Hz,1H),7.34–7.22(m,6H),7.21–7.14(m,1H),5.68(s,1H),4.45(q,J＝7.0Hz,1H),3.28(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.84,180.95,169.15,159.21,156.80,147.55,139.35,136.81,134.90,133.42,132.63,132.48,130.17,129.22,129.22,128.30,128.30,126.72,125.98,125.37,116.97,107.54,101.07,57.62,36.98。HR-MS(m/z)(ESI)：计算为C 25H 19BrFN 2O 3[M+H] +:493.0558；实测值：493.0592。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二氟苯基)-丙酰胺(IIw)
Figure PCTCN2019095516-appb-000038
产率：65.8％。黄色固体。熔点168.4～169.2℃。 1H NMR(400MHz,DMSO):δ10.50(s,1H),7.98–7.94(m,1H),7.89(dd,J＝7.6,1.0Hz,1H),7.78(td,J＝7.5,1.3Hz,1H),7.70(td,J＝7.5,1.3Hz,1H),7.33–7.24(m,7H),7.22–7.16(m,1H),6.96–6.89(m,1H),5.68(s,1H),4.45(q,J＝7.0Hz,1H),3.28(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.88,180.95,169.34,163.56,161.14,147.57,140.74,136.80,134.91,132.63,132.50,130.18,129.23,129.23,128.32,128.32,126.74,125.99,125.38,102.69,102.40,101.12,99.05,57.67,36.94。HR-MS(m/z)(ESI)：计算为C 25H 19F 2N 2O 3[M+H] +:433.1358；实测值：433.1345。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟-4-甲基苯基)-丙酰胺(IIx)
Figure PCTCN2019095516-appb-000039
产率：70.5％。黄色固体。熔点158.5～158.7℃。 1H NMR(400MHz,DMSO):δ9.95(s,1H),7.96(dd,J＝7.7,1.0Hz,1H),7.90(dd,J＝7.7,1.0Hz,1H),7.79(td,J＝7.5,1.3Hz,1H),7.70(td,J＝7.5,1.3Hz,1H),7.60(t,J＝8.3Hz,1H),7.39–7.32(m,2H),7.27(t,J＝7.5Hz,2H),7.19(ddd,J＝7.2,6.6,3.4Hz,2H),7.08(dd,J＝11.8,1.1Hz,1H),6.96(d,J＝8.1Hz,1H),5.73(s,1H),4.60(dt,J＝14.0,7.1Hz,1H),3.34–3.20(m,2H),2.27(s,3H)。 13C NMR(101MHz,DMSO):δ181.83,181.00,169.03,155.13,152.70,147.51,136.92,134.89,132.67,132.45,130.18,129.29,129.29,128.25,128.25,126.66,125.96,125.36,124.80,124.49,122.55,122.43,101.07,57.11,37.22,20.35。HR-MS(m/z)(ESI)：计算为C 26H 22FN 2O 3Na[M+Na] +:451.1428；实测值：451.1410。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2,5-二甲氧基苯基)-丙酰胺(IIy)
Figure PCTCN2019095516-appb-000040
产率：71.2％。黄色固体。熔点117.7～117.9℃。 1H NMR(400MHz,DMSO):δ9.52(s,1H),7.97(dd,J＝7.7,1.0Hz,1H),7.90(dd,J＝7.6,1.0Hz,1H),7.81(td,J＝7.5,1.3Hz,1H),7.72(td,J＝7.5,1.4Hz,1H),7.59(d,J＝3.1Hz,1H),7.33(d,J＝7.1Hz,2H),7.25(dd,J＝16.1,8.2Hz,3H),7.21–7.16(m,1H),6.96(d,J＝9.0 Hz,1H),6.65(dd,J＝8.9,3.1Hz,1H),5.78(s,1H),4.76(dt,J＝14.0,7.0Hz,1H),3.74(s,3H),3.67(s,3H),3.31–3.18(m,2H)。 13C NMR(101MHz,DMSO):δ181.86,181.08,168.98,152.92,147.49,143.87,137.00,134.96,132.67,132.49,130.17,129.31,129.31,128.22,128.22,127.35,126.61,125.99,125.37,112.22,108.84,108.48,101.24,57.13,56.40,55.36,37.28。HR-MS(m/z)(ESI)：计算为C 27H 24N 2O 5Na[M+Na] +:479.1577；实测值：479.1561。
(R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯-4-甲基苯基)-丙酰胺(IIz)
Figure PCTCN2019095516-appb-000041
产率：71.2％。黄色固体。熔点：178.1～178.6℃。 1H NMR(400MHz,DMSO):δ10.22(s,1H),7.97(dd,J＝7.6,0.9Hz,1H),7.89(dd,J＝7.6,1.0Hz,1H),7.80(td,J＝7.5,1.3Hz,1H),7.72(ddd,J＝8.9,5.6,1.3Hz,2H),7.29(tdd,J＝16.3,8.6,3.7Hz,7H),7.18(ddd,J＝7.0,3.8,1.4Hz,1H),5.68(s,1H),4.43(q,J＝7.0Hz,1H),3.27(d,J＝6.8Hz,2H),2.26(s,3H)。 13C NMR(101MHz,DMSO):δ181.80,180.95,168.70,147.56,137.34,136.91,134.90,132.99,132.64,132.47,131.24,130.58,130.18,129.22,129.22,128.28,128.28,126.67,125.97,125.36,119.66,118.30,100.98,57.57,37.03,18.94。HR-MS(m/z)(ESI)：计算为C 26H 22ClN 2O 3[M+H] +:445.1313；实测值：475.1306。
实施例2式(I)的化合物(化合物Ia-Iz)(2-氨基酸酰胺-肟基萘醌类化合物)的合成
将如上合成的1,4-萘醌-2-氨基酰化的取代苯胺类化合物与盐酸羟胺以质量比为1:3加入装有25mL无水乙醇的圆底烧瓶中，控制温度在80℃下回流反应12h，减压旋转干燥，用水和二氯甲烷萃取三次(3×30mL)，用无水硫酸钠干燥2h，减压旋转干燥，经洗脱剂为石油醚:乙酸乙酯(5:1)的硅胶柱纯化，旋转干燥得到的产品为黄绿色固体。所得产物具有如下具体结构式和参数表征。
式(I)的化合物的具体结构式及参数表征如下：
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-(三氟甲基)苯基)-丙酰胺(Ia)
Figure PCTCN2019095516-appb-000042
产率：48.2％。黄绿色固体。熔点194.4～194.7℃。 1H NMR(400MHz,DMSO):δ12.33(s,1H),10.57(s,1H),8.16(dd,J＝8.0,0.6Hz,1H),8.06(dd,J＝7.9,1.0Hz,1H),7.78(d,J＝8.6Hz,2H),7.67(dd,J＝11.2,5.2Hz,3H),7.60–7.54(m,1H),7.35–7.24(m,4H),7.20(ddd,J＝6.7,3.9,1.7Hz,1H),6.60(s,1H),6.36(d,J＝8.2Hz,1H),4.43(q,J＝7.0Hz,1H),3.24(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.84,170.34,145.21,141.99,140.42,136.99,133.85,132.93,129.21,129.21,128.99,128.42,128.33,128.33,126.69,126.18,126.14,125.83,123.94,123.62,122.32,119.49,119.49,91.42,57.88,37.46。HR-MS(m/z)(ESI)：计算为C 26H 21F 3N 3O 3[M+H] +:480.1530；实测值：480.1521。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟苯基)-丙酰胺(Ib)
Figure PCTCN2019095516-appb-000043
产率：55.3％。黄绿色固体。熔点226.9～227.2℃。 1H NMR(400MHz,DMSO):δ12.38(s,1H),10.09(s,1H),8.16(d,J＝7.8Hz,1H),8.04(d,J＝7.1Hz,1H),7.77(td,J＝7.8,2.9Hz,1H),7.70–7.65(m,1H),7.60–7.53(m,1H),7.28(dt,J＝9.5, 4.4Hz,5H),7.18(ddd,J＝9.7,8.9,3.2Hz,3H),6.64(s,1H),6.33(d,J＝8.3Hz,1H),4.57(dd,J＝14.6,7.1Hz,1H),3.21(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ180.05,170.36,155.27,152.83,145.41,140.57,137.21,133.99,133.11,129.41,129.41,129.16,128.54,128.45,128.45,126.82,125.97,125.50,124.59,122.48,115.87,115.68,91.65,57.42,37.71。HR-MS(m/z)(ESI)：计算为C 25H 20FN 3O 3Na[M+Na] +:452.1381；实测值：452.1380。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3,N-二苯基-丙酰胺(Ic)
Figure PCTCN2019095516-appb-000044
产率：50.6％。黄绿色固体。熔点229.6～230.2℃。 1H NMR(400MHz,DMSO):δ10.27(s,1H),8.12(d,J＝7.9Hz,1H),8.05–7.97(m,1H),7.70–7.60(m,1H),7.61–7.52(m,1H),7.50(d,J＝7.7Hz,2H),7.34–7.22(m,7H),7.18(td,J＝5.7,2.5Hz,1H),7.06(t,J＝7.4Hz,1H),6.58(s,1H),6.25(d,J＝8.0Hz,1H),4.35(dd,J＝13.6,7.8Hz,1H),3.23–3.12(m,2H)。 13C NMR(101MHz,DMSO):δ180.44,170.28,145.80,140.88,138.63,137.39,134.21,133.59,129.67,129.67,129.63,129.38,129.38,128.89,128.79,128.79,127.28,126.36,124.56,122.83,120.28,120.28,91.85,58.27,38.02。HR-MS(m/z)(ESI)：计算为C 25H 21N 3O 3Na[M+Na] +:434.1475；实测值：434.1469。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟苯基)-丙酰胺(Id)
Figure PCTCN2019095516-appb-000045
产率：49.9％。黄绿色固体。熔点202.2～203.1℃。 1H NMR(400MHz,DMSO):δ12.37(s,1H),10.45(s,1H),8.19–8.14(m,1H),8.05(dd,J＝7.9,0.8Hz,1H),7.71–7.63(m,1H),7.60–7.52(m,2H),7.36(dd,J＝11.4,4.7Hz,1H),7.33–7.25(m,6H),7.23–7.16(m,1H),6.90(td,J＝8.2,1.8Hz,1H),6.60(s,1H),6.35(d,J＝8.2Hz,1H),4.40(q,J＝7.0Hz,1H),3.22(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.89,170.13,163.36,160.96,145.27,140.48,140.23,140.12,137.11,133.89,132.98,130.61,129.26,129.03,128.47,128.39,126.74,125.89,122.37,115.38,110.42,106.55,91.44,57.90,37.55。HR-MS(m/z)(ESI)：计算为C 25H 20FN 3O 3Na[M+Na] +:452.1381；实测值：452.1377。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲氧基苯基)-丙酰胺(Ie)
Figure PCTCN2019095516-appb-000046
产率：54.8％。黄色固体。熔点176.9～177.4℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),9.49(s,1H),8.17(dd,J＝8.0,0.6Hz,1H),8.06(dd,J＝7.9,1.0Hz,1H),7.88(dd,J＝8.0,1.5Hz,1H),7.71–7.64(m,1H),7.61–7.51(m,1H),7.34–7.31(m,2H),7.26(t,J＝7.4Hz,2H),7.21–7.16(m,1H),7.11–7.05(m,1H),7.02 (dd,J＝8.2,1.3Hz,1H),6.93–6.88(m,1H),6.65(s,1H),6.41(d,J＝8.3Hz,1H),4.63(dd,J＝14.3,7.4Hz,1H),3.76(s,3H),3.21(d,J＝6.9Hz,2H)。 13C NMR(101MHz,DMSO):δ179.92,169.74,149.85,145.27,140.48,137.34,133.91,132.92,129.30,129.30,128.98,128.44,128.25,128.25,126.65,126.56,125.82,124.91,122.35,122.00,120.33,111.38,91.65,57.43,55.80,37.58。HR-MS(m/z)(ESI)：计算为C 26H 24N 3O 4[M+H] +:442.1761；实测值：442.1748。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲基苯基)-丙酰胺(If)
Figure PCTCN2019095516-appb-000047
产率：52.5％。黄绿色固体。熔点128.3～129.2℃。 1H NMR(400MHz,DMSO):δ12.34(s,1H),10.06(s,1H),8.16(d,J＝7.6Hz,1H),8.05(dd,J＝7.9,1.0Hz,1H),7.72–7.64(m,1H),7.60–7.54(m,1H),7.31–7.25(m,4H),7.21–7.17(m,3H),6.64(d,J＝49.4Hz,2H),6.30(d,J＝8.2Hz,1H),4.36(dd,J＝14.2,7.4Hz,1H),3.20–3.16(m,2H),2.22(s,6H)。 13C NMR(101MHz,DMSO):δ179.94,169.58,145.30,140.50,138.35,137.89,137.89,137.24,133.91,133.00,129.27,129.27,129.06,128.47,128.39,128.39,126.71,125.89,125.33,122.39,117.37,117.37,91.40,57.90,37.67,21.11,21.11。HR-MS(m/z)(ESI)：计算为C 27H 25N 3O 3Na[M+Na] +:462.1788；实测值：462.1788。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氯苯基)-丙酰胺(Ih)
Figure PCTCN2019095516-appb-000048
产率：53.2％。黄绿色固体。熔点192.3～193.0℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.36(s,1H),8.17(d,J＝7.9Hz,1H),8.05(dd,J＝7.9,0.9Hz,1H),7.70–7.63(m,1H),7.61–7.54(m,3H),7.40–7.34(m,2H),7.32–7.24(m,4H),7.19(dt,J＝9.2,4.2Hz,1H),6.61(s,1H),6.33(d,J＝8.1Hz,1H),4.40(dd,J＝14.5,7.0Hz,1H),3.22(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.89,169.85,145.27,140.45,137.39,137.09,133.89,132.92,129.24,129.24,128.98,128.75,128.75,128.46,128.35,128.35,127.43,126.69,125.85,122.35,121.21,121.21,91.43,57.83,37.60。HR-MS(m/z)(ESI)：计算为C 25H 20ClN 3O 3Na[M+Na] +:468.1085；实测值：468.1084。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-溴苯基)-丙酰胺(Ii)
Figure PCTCN2019095516-appb-000049
产率：54.1％。黄绿色固体。熔点179.2～179.4℃. 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.35(s,1H),8.16(d,J＝7.9Hz,1H),8.09–8.01(m,1H),7.73–7.61(m,1H),7.62–7.47(m,5H),7.28(t,J＝7.3Hz,4H),7.19(dt,J＝9.2,4.2Hz,1H),6.60(s,1H),6.33(d,J＝8.1Hz,1H),4.39(q,J＝7.0Hz,1H),3.22(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.91,169.90,145.29,140.46,137.82,137.09,133.90,132.97,131.69,131.69,129.26,129.26,129.03,128.47,128.38, 128.38,126.73,125.89,122.37,121.60,121.60,115.49,91.43,57.88,37.59。HR-MS(m/z)(ESI)：计算为C 25H 20BrN 3O 3Na[M+Na] +:512.0580；实测值：512.0580。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴-4-氟苯基)-丙酰胺(Ij)
Figure PCTCN2019095516-appb-000050
产率：55.5％。黄绿色固体。熔点166.9～170.6℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.53(s,1H),8.16(d,J＝7.8Hz,1H),8.05(d,J＝7.0Hz,1H),7.72–7.55(m,4H),7.28(d,J＝6.8Hz,5H),7.20(dd,J＝5.9,2.6Hz,1H),6.58(s,1H),6.35(d,J＝8.1Hz,1H),4.38(q,J＝7.0Hz,1H),3.22(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.82,170.25,159.24,156.83,145.20,140.42,139.61,136.98,133.83,133.46,132.94,129.21,129.21,129.00,128.42,128.35,126.71,125.84,122.32,116.92,107.68,101.47,91.41,57.90,37.44。HR-MS(m/z)(ESI)：计算为C 25H 19BrFN 3O 3Na[M+Na] +:530.0486；实测值：530.0488。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺(Ik)
Figure PCTCN2019095516-appb-000051
产率：38.9％。黄绿色固体。熔点118.9～119.5℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.54(s,1H),8.16(d,J＝7.8Hz,1H),8.10–7.98(m,2H),7.78(d,J＝8.3Hz,1H),7.71–7.64(m,1H),7.60–7.53(m,2H),7.43(d,J＝8.2Hz,1H),7.35–7.25(m,4H),7.20(t,J＝7.0Hz,1H),6.60(s,1H),6.36(d,J＝8.1Hz,1H),4.40(dd,J＝14.4,7.1Hz,1H),3.30(s,1H),3.24(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ179.85,170.32,145.23,140.48,139.19,137.08,133.86,132.95,130.16,129.23,129.23,129.01,128.45,128.37,128.37,126.72,125.85,125.42,123.18,122.34,120.18,115.69,115.65,91.42,57.95,37.50。HR-MS(m/z)(ESI)：计算为C 26H 20F 3N 3O 3Na[M+Na] +:502.1349；实测值：502.1341。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氟苯基)-丙酰胺(Il)
Figure PCTCN2019095516-appb-000052
产率：56.6％。黄绿色固体。熔点175.3～175.9℃。 1H NMR(400MHz,DMSO):δ12.36(s,1H),10.35(s,1H),8.16(d,J＝7.8Hz,1H),8.05(d,J＝7.1Hz,1H),7.74–7.64(m,1H),7.61–7.51(m,4H),7.34–7.25(m,5H),7.17(dt,J＝17.7,7.9Hz,4H),6.61(s,1H),6.32(d,J＝5.4Hz,1H),4.39(d,J＝4.7Hz,1H),3.22(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ179.93,169.64,159.56,157.17,145.30,140.48,137.19,134.88,134.85,133.93,132.96,129.29,129.02,128.49,128.38,126.72,125.89,122.39,121.58,121.50,115.56,115.34,91.43,57.79,37.66。HR-MS(m/z)(ESI)：计算为C 25H 20FN 3O 3Na[M+Na] +:452.1381；实测值：452.1381。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲苯基)-丙酰胺(Im)
Figure PCTCN2019095516-appb-000053
产率：49.0％。黄绿色固体。熔点116.6～117.6℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.15(s,1H),8.16(d,J＝7.6Hz,1H),8.05(dd,J＝7.9,0.9Hz,1H),7.73–7.63(m,1H),7.64–7.48(m,1H),7.42–7.25(m,6H),7.24–7.14(m,2H),6.88(d,J＝7.5Hz,1H),6.60(s,1H),6.31(d,J＝8.2Hz,1H),4.38(dd,J＝14.5,7.2Hz,1H),3.20(d,J＝6.9Hz,2H),2.26(s,3H)。 13C NMR(101MHz,DMSO):δ179.94,169.64,145.31,140.50,138.42,138.11,137.23,133.92,133.00,129.28,129.28,129.05,128.73,128.48,128.39,128.39,126.71,125.90,124.53,122.39,120.19,116.84,91.41,57.86,37.67,21.20。HR-MS(m/z)(ESI)：计算为C 26H 24N 3O 3[M+H] +:426.1812；实测值：426.1810。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氯苯基)-丙酰胺(In)
Figure PCTCN2019095516-appb-000054
产率：55.9％。黄绿色固体。熔点129.7～130.6℃。 1H NMR(400MHz,DMSO):δ12.36(s,1H),9.86(s,1H),8.17(d,J＝8.0Hz,1H),8.05(dd,J＝7.9,0.9Hz,1H),7.71–7.65(m,1H),7.62(dd,J＝8.0,1.4Hz,1H),7.59–7.54(m,1H),7.48(dd,J＝8.0,1.3Hz,1H),7.32(ddd,J＝9.9,7.3,5.4Hz,5H),7.20(ddd,J＝9.2,3.5,1.5Hz, 2H),6.68(s,1H),6.39(d,J＝8.1Hz,1H),4.55(dd,J＝14.5,7.1Hz,1H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ179.92,170.18,145.28,140.51,137.21,134.30,133.92,132.97,129.57,129.33,129.33,129.03,128.47,128.39,128.39,127.54,126.96,126.83,126.72,126.29,125.86,122.38,91.89,57.52,37.54。HR-MS(m/z)(ESI)：计算为C 25H 21ClN 3O 3Na[M+H] +:446.1266；实测值：446.1267。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯苯基)-丙酰胺(Io)
Figure PCTCN2019095516-appb-000055
产率：48.6％。黄绿色固体。熔点159.1～159.9℃。 1H NMR(400MHz,DMSO):δ12.36(s,1H),10.42(s,1H),8.16(d,J＝7.9Hz,1H),8.05(d,J＝7.6Hz,1H),7.75(s,1H),7.67(dd,J＝11.2,4.0Hz,1H),7.57(t,J＝7.3Hz,1H),7.43(d,J＝8.5Hz,1H),7.37–7.25(m,5H),7.19(t,J＝6.6Hz,1H),7.13(dd,J＝7.9,0.9Hz,1H),6.59(s,1H),6.33(d,J＝8.1Hz,1H),4.39(dd,J＝14.4,7.0Hz,1H),3.22(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.87,170.11,145.24,140.45,139.87,137.06,133.87,133.14,132.96,130.59,129.24,129.24,129.02,128.45,128.37,128.37,126.72,125.86,123.53,122.35,119.10,118.02,91.44,57.89,37.50。HR-MS(m/z)(ESI)：计算为C 25H 21ClN 3O 3[M+H] +:446.1266；实测值：446.1252。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-溴苯基)-丙酰胺(Ip)
Figure PCTCN2019095516-appb-000056
产率：57.2％。黄绿色固体。熔点361.2～361.3℃。 1H NMR(400MHz,DMSO):δ12.30(s,1H),9.75(s,1H),8.11(dd,J＝8.0,0.6Hz,1H),7.99(dd,J＝7.9,1.0Hz,1H),7.65–7.56(m,2H),7.50(ddd,J＝9.6,7.0,1.4Hz,2H),7.32–7.27(m,3H),7.23(t,J＝7.5Hz,2H),7.18–7.12(m,1H),7.08(td,J＝7.8,1.6Hz,1H),6.61(s,1H),6.33(d,J＝8.1Hz,1H),4.44(dd,J＝13.8,7.8Hz,1H),3.25–3.19(m,2H)。 13C NMR(101MHz,DMSO):δ179.91,170.10,145.28,140.55,137.27,135.63,133.92,132.98,132.76,129.32,129.32,129.04,128.47,128.42,128.42,128.17,127.45,126.94,126.73,125.86,122.39,117.95,91.96,57.60,37.48。HR-MS(m/z)(ESI)：计算为C 25H 21BrN 3O 3[M+H] +:490.0761；实测值：490.0744。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺(Iq)
Figure PCTCN2019095516-appb-000057
产率：51.1％。黄绿色固体。熔点129.8～130.3℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.38(s,1H),8.16(d,J＝7.5Hz,1H),8.05(dd,J＝7.9,1.0Hz,1H),7.88(d,J＝1.8Hz,1H),7.68(td,J＝7.8,1.4Hz,1H),7.62–7.53(m,1H),7.47(dt,J＝7.1,2.1Hz,1H),7.34–7.25(m,6H),7.20(ddt,J＝8.5,5.5,2.9Hz,1H),6.58(s, 1H),6.33(d,J＝8.1Hz,1H),4.37(dd,J＝14.5,7.1Hz,1H),3.22(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ179.88,170.10,145.25,140.47,140.01,137.06,133.87,132.99,130.91,129.24,129.24,129.05,128.45,128.39,128.39,126.74,126.45,125.88,122.36,121.96,121.60,118.41,91.44,57.91,37.51。HR-MS(m/z)(ESI)：计算为C 25H 21BrN 3O 3[M+H] +:490.0761；实测值：490.0760。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺(Ir)
Figure PCTCN2019095516-appb-000058
产率：50.6％。黄绿色固体。熔点112.3～113.2℃。 1H NMR(400MHz,DMSO):δ12.34(s,1H),10.21(s,1H),8.16(d,J＝7.5Hz,1H),8.05(dd,J＝7.9,1.0Hz,1H),7.71–7.65(m,1H),7.60–7.54(m,1H),7.32–7.18(m,7H),7.12–7.07(m,1H),6.65(dd,J＝8.2,1.9Hz,1H),6.60(s,1H),6.30(d,J＝8.2Hz,1H),4.38(dd,J＝14.3,7.3Hz,1H),3.72(s,3H),3.20(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ179.90,169.71,159.56,145.26,140.45,139.62,137.17,133.88,132.96,129.67,129.24,129.24,129.02,128.45,128.35,128.35,126.68,125.86,122.35,111.89,109.29,105.38,91.40,57.83,55.05,37.63。HR-MS(m/z)(ESI)：计算为C 26H 24N 3O 4[M+H] +:442.1761；实测值：442.1760。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲氧基苯基)-丙酰胺(Is)
Figure PCTCN2019095516-appb-000059
产率：55.7％。黄绿色固体。熔点89.9～90.5℃。 1H NMR(400MHz,DMSO):δ12.33(s,1H),10.06(s,1H),8.16(d,J＝7.6Hz,1H),8.05(dd,J＝7.9,1.0Hz,1H),7.73–7.64(m,1H),7.62–7.53(m,1H),7.51–7.38(m,2H),7.33–7.26(m,4H),7.20(dd,J＝5.8,2.8Hz,1H),6.97–6.82(m,2H),6.59(s,1H),6.28(d,J＝8.1Hz,1H),4.35(dd,J＝14.4,7.0Hz,1H),3.19(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.91,169.12,155.64,145.28,140.44,137.22,133.90,132.95,131.50,129.25,129.25,129.01,128.46,128.34,128.34,126.66,125.86,122.35,121.30,121.30,113.96,113.96,91.35,57.67,55.23,37.69。HR-MS(m/z)(ESI)：计算为C 26H 24N 3O 4[M+H] +:442.1761；实测值：442.1761。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲苯基)-丙酰胺(It)
Figure PCTCN2019095516-appb-000060
产率：50.5％。黄绿色固体。熔点137.2～137.4℃。 1H NMR(400MHz,DMSO):δ12.36(s,1H),10.24(s,1H),8.16(d,J＝7.6Hz,1H),8.05(dd,J＝7.9,0.9Hz,1H),7.70–7.64(m,1H),7.59–7.53(m,1H),7.44(d,J＝8.4Hz,2H),7.33–7.23(m,5H),7.21–7.16(m,1H),7.10(d,J＝8.3Hz,2H),6.60(s,1H),6.29(d,J＝8.2Hz,1H),4.40(dd,J＝14.1,7.4Hz,1H),3.29–3.12(m,3H),2.24(s,4H)。 13C NMR(101MHz,DMSO):δ179.90,169.39,145.24,140.41,137.22,135.98,133.91,132.92,132.74,129.26,129.26,129.17,129.17,128.97,128.44,128.31,128.31,126.63, 125.84,122.35,119.67,119.67,91.36,57.68,37.67,20.50。HR-MS(m/z)(ESI)：计算为C 26H 23N 3O 3Na[M+Na] +:448.1632；实测值：448.1619。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲氧基苯基)-丙酰胺(Iu)
Figure PCTCN2019095516-appb-000061
产率：50.5％。黄绿色固体。熔点114.1～114.6℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.19(s,1H),8.16(d,J＝7.9Hz,1H),8.10–8.02(m,1H),7.72–7.63(m,1H),7.56(dd,J＝11.1,4.0Hz,1H),7.38–7.25(m,4H),7.20(dt,J＝9.1,4.2Hz,1H),6.81(d,J＝2.2Hz,2H),6.59(s,1H),6.30(d,J＝8.2Hz,1H),6.24(t,J＝2.2Hz,1H),4.37(dd,J＝14.1,7.5Hz,1H),3.71(s,6H),3.26–3.14(m,2H)。 13C NMR(101MHz,DMSO):δ179.89,169.77,160.55,160.55,145.26,140.46,140.12,137.17,133.88,132.95,129.23,129.23,129.01,128.45,128.35,128.35,126.68,125.86,122.35,97.89,97.89,95.82,91.41,57.89,55.16,55.16,37.62。HR-MS(m/z)(ESI)：计算为C 27H 26N 3O 5[M+H] +:472.1867；实测值：472.1851。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟-4-溴苯基)-丙酰胺(Iv)
Figure PCTCN2019095516-appb-000062
产率：49.2％。黄绿色固体。熔点121.2～122.2℃。 1H NMR(400MHz,DMSO):δ10.47(s,1H),7.99(dd,J＝7.6,0.9Hz,1H),7.90(dd,J＝7.6,1.0Hz,1H),7.82(td,J＝7.5,1.3Hz,1H),7.76–7.62(m,3H),7.36–7.15(m,7H),5.67(s,1H),4.44(q,J＝7.0Hz,1H),3.27(d,J＝6.8Hz,2H)。 13C NMR(101MHz,DMSO):δ181.89,181.00,169.19,147.63,139.47,139.37,136.85,135.01,133.50,132.66,132.59,130.22,129.25,128.34,126.75,126.06,125.42,117.05,107.82,107.55,101.64,101.44,101.06,57.64,36.96。HR-MS(m/z)(ESI)：计算为C 25H 19BrFN 3O 3Na[M+Na] +:530.0486；实测值：530.0471。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二氟苯基)-丙酰胺(Iw)
Figure PCTCN2019095516-appb-000063
产率：43.5％。黄绿色固体。熔点163.3～163.6℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),10.57(s,1H),8.20–8.15(m,1H),8.05(dd,J＝7.9,1.0Hz,1H),7.68(td,J＝7.8,1.4Hz,1H),7.61–7.53(m,1H),7.37–7.26(m,6H),7.21(dd,J＝5.9,2.8Hz,1H),6.93(tt,J＝9.3,2.3Hz,1H),6.57(s,1H),6.34(d,J＝8.1Hz,1H),4.38(dd,J＝14.5,7.0Hz,1H)。 13C NMR(101MHz,DMSO):δ179.82,170.43, 163.72,161.30,145.21,140.87,140.43,136.93,133.83,132.95,129.20,129.20,129.01,128.42,128.36,128.36,126.73,125.84,122.32,102.59,102.30,98.96,91.46,57.95,37.40。HR-MS(m/z)(ESI)：计算为C 25H 19F 2N 3O 3Na[M+Na] +:470.1287；实测值：470.1270。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟-4-甲基苯基)-丙酰胺(Ix)
Figure PCTCN2019095516-appb-000064
产率：60.2％。黄绿色固体。熔点218.5～219.4℃。 1H NMR(400MHz,DMSO):δ9.96(s,1H),8.16(dd,J＝8.0,0.6Hz,1H),8.05(dd,J＝7.9,1.0Hz,1H),7.72–7.65(m,1H),7.58(dt,J＝8.3,4.7Hz,2H),7.30(dt,J＝14.9,4.6Hz,4H),7.23–7.17(m,1H),7.08(dd,J＝11.8,1.1Hz,1H),6.96(d,J＝8.2Hz,1H),6.63(s,1H),6.32(d,J＝8.3Hz,1H),4.55(q,J＝6.9Hz,1H),3.21(d,J＝6.7Hz,2H),2.27(s,3H)。 13C NMR(101MHz,DMSO):δ179.91,170.03,155.08,152.64,145.27,140.44,137.15,136.01,133.91,132.94,129.30,128.99,128.45,128.30,126.65,125.84,124.83,124.80,124.38,122.63,116.05,115.86,91.49,57.24,37.63,20.41。HR-MS(m/z)(ESI)：计算为C 26H 23FN 3O 3[M+H] +:444.1718；实测值：444.1704。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2,5-二甲氧基苯基)-丙酰胺(Iy)
Figure PCTCN2019095516-appb-000065
产率：49.3％。黄绿色固体。熔点121.1～121.6℃。 1H NMR(400MHz,DMSO):δ12.35(s,1H),9.48(s,1H),8.17(d,J＝7.6Hz,1H),8.06(dd,J＝7.9,0.8Hz,1H),7.72–7.61(m,2H),7.61–7.54(m,1H),7.32(d,J＝7.1Hz,2H),7.26(t,J＝7.4Hz,2H),7.18(t,J＝7.2Hz,1H),6.94(d,J＝9.0Hz,1H),6.63(dd,J＝8.3,3.7Hz,2H),6.43(d,J＝8.3Hz,1H),4.65(dd,J＝14.5,7.1Hz,1H),3.70(s,3H),3.67(s,3H),3.21(d,J＝6.7Hz,2H)。 13C NMR(101MHz,DMSO):δ179.90,169.86,152.99,145.24,143.70,140.47,137.32,133.89,132.92,129.28,129.28,128.97,128.42,128.24,128.24,126.55,125.82,122.34,112.28,108.62,108.14,91.64,57.42,56.44,55.37,37.52。HR-MS(m/z)(ESI)：计算为C 27H 26N 3O 5[M+H] +:472.1867；实测值：472.1851。
(R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯-4-甲基苯基)-丙酰胺(Iz)
Figure PCTCN2019095516-appb-000066
产率：51.2％。黄绿色固体。熔点169.1～169.5℃。 1H NMR(400MHz,DMSO):δ12.28(s,1H),10.30(s,1H),8.16(d,J＝7.5Hz,1H),8.05(dd,J＝7.9,1.0Hz,1H),7.72(d,J＝2.0Hz,1H),7.70–7.65(m,1H),7.60–7.54(m,1H),7.33(dd,J＝8.3, 2.1Hz,1H),7.30–7.26(m,4H),7.20(td,J＝5.7,2.6Hz,1H),6.65–6.64(m,1H),6.58(s,1H),6.31(d,J＝8.1Hz,1H),4.36(dd,J＝14.5,7.0Hz,1H),3.21(d,J＝6.7Hz,3H),2.26(s,3H)。 13C NMR(101MHz,DMSO):δ179.85,169.80,145.22,140.42,137.52,137.06,133.86,133.02,132.94,131.27,130.49,129.21,129.21,129.00,128.43,128.34,128.34,126.69,125.84,122.33,119.59,118.25,91.40,57.81,37.52,18.96。HR-MS(m/z)(ESI)：计算为C 26H 23ClN 3O 3[M+H] +:460.1422；实测值：460.1405。
实施例3式(III)的化合物(2-氨基酸酰胺-羟肟酯基萘醌类化合物)的合成
在氩气保护下，取化合物Ik、化合物Iq、化合物Ir各0.5g分别与4-叔丁基苯磺酰氯按物质的量1:1.5的比例溶于10mL的二氯甲烷中，在冰浴条件下搅拌30min，随后逐滴加入物质的量是肟类化合物2倍的三乙胺，随后搅拌10min，再用20mL的冰水淬灭反应。用10mL的二氯甲烷萃取三次，合并有机层溶液，用无水硫酸钠干燥2h，减压旋转干燥，用洗脱剂为石油醚:乙酸乙酯(8:1)的硅胶柱纯化，旋转干燥得到的最终产品为黄色固体。所得产物(化合物IIIk-1、IIIq和IIIr)的具体结构式和参数表征如下。
在氩气保护下，取0.5g化合物Ik与2-噻吩-苯磺酰氯按照1:1.5的比例溶于10ml的二氯甲烷中，在冰浴条件下搅拌30min，随后逐滴加入物质的量是肟类化合物2倍的三乙胺，滴完后搅拌10min，再用20mL的冰水淬灭反应。用10ml的二氯甲烷萃取三次，合并有机层溶液，用无水硫酸钠干燥2h，减压旋转干燥，用洗脱剂为石油醚:乙酸乙酯(8:1)的硅胶柱纯化，旋转干燥得到的最终产品为黄色固体。所得产物(化合物IIIk-2)的具体结构式和参数表征如下。
式(III)的化合物的具体结构式及参数表征如下：
(R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺(IIIk-1)
Figure PCTCN2019095516-appb-000067
产率：80.1％。黄色固体。熔点118.5～119.2℃。 1H NMR(400MHz,DMSO):δ10.71(s,1H),8.07(s,1H),7.98(dd,J＝7.7,1.3Hz,1H),7.95–7.92(m,1H),7.84(dd,J＝15.3,8.5Hz,3H),7.66–7.53(m,5H),7.46(d,J＝7.8Hz,1H),7.34(d,J＝7.0Hz,2H),7.25(t,J＝7.5Hz,2H),7.16(t,J＝7.3Hz,2H),6.12(s,1H),4.56(dd,J＝13.8,7.7Hz,1H),3.32–3.18(m,2H),1.22(d,J＝11.2Hz,9H)。 13C NMR(101MHz,DMSO):δ179.14,169.46,157.86,151.72,143.46,139.06,136.92,134.02,131.98,131.12,130.19,129.30,129.30,129.00,128.33,128.33,128.28,128.28,126.73,126.32,126.32,125.34,124.34,123.16,122.69,120.30,115.72,88.16,57.85,37.45,34.97,31.04,30.52,30.52,30.52,26.32。HR-MS(m/z)(ESI):计算为C 36H 33N 3O 5S[M+H] +:676.2088；实测值：676.2062。
(R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺(IIIq)
Figure PCTCN2019095516-appb-000068
产率：85.2％。黄色固体。熔点125.3～125.6℃。 1H NMR(400MHz,DMSO):δ10.54(s,1H),8.01(dd,J＝7.7,1.3Hz,1H),7.96–7.92(m,2H),7.88–7.80(m,2H),7.69(td,J＝7.6,1.5Hz,1H),7.63(td,J＝7.6,1.2Hz,1H),7.60–7.56(m,2H),7.49(dt,J＝7.2,2.0Hz,1H),7.35–7.30(m,4H),7.25(t,J＝7.5Hz,2H),7.15(dd,J＝17.3,7.7Hz,2H),6.09(s,1H),4.51(dd,J＝14.0,7.5Hz,1H),3.32–3.15(m,2H),1.23(s,10H)。 13C NMR(101MHz,DMSO):δ179.21,169.27,157.98,151.80,143.49,139.86,136.90,134.18,131.98,131.28,131.17,130.98,129.34,129.34,129.07,128.40,128.40,128.33,126.80,126.66,126.44,126.44,125.36,124.39,123.23,122.04,121.70,118.51,88.20,57.85,37.47,35.09,30.65,30.65,30.65。HR-MS(m/z)(ESI):计算为C 35H 32BrN 3O 5S[M+Na] +:708.1138；实测值：708.1109。
(R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3- 苯基-N-(3-甲氧基苯基)-丙酰胺(IIIr)
Figure PCTCN2019095516-appb-000069
产率：85.5％。黄色固体。熔点104.2～104.9℃。 1H NMR(400MHz,DMSO):δ10.41(s,1H),7.95(dd,J＝7.6,1.4Hz,1H),7.89(dd,J＝13.0,5.0Hz,3H),7.61–7.52(m,4H),7.36–7.30(m,4H),7.26(dt,J＝7.7,6.1Hz,4H),7.17(d,J＝7.6Hz,2H),7.09(d,J＝8.1Hz,1H),6.70(dd,J＝8.2,1.8Hz,1H),6.11(s,1H),4.54(dd,J＝14.0,7.5Hz,1H),3.74(s,4H),3.30–3.21(m,2H),1.22(d,J＝11.7Hz,12H)。 13C NMR(101MHz,DMSO):δ179.15,168.88,159.60,157.85,151.76,143.39,139.53,137.00,133.99,132.00,131.13,131.09,129.73,129.32,129.32,128.96,128.31,128.31,126.69,126.62,126.37,126.37,125.35,124.38,123.15,111.88,109.36,105.39,88.10,57.76,55.01,34.99,31.05,30.56,30.56,30.56。HR-MS(m/z)(ESI):计算为C 36H 36N 3O 6S[M+H] +:638.2319；实测值：638.2294。
(R)-2-((4-(((噻吩-2-基磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺(IIIk-2)
Figure PCTCN2019095516-appb-000070
产率：80.2％。黄色固体。熔点163.4～164.8℃。 1H NMR(400MHz,MeOD):δ 8.03–7.95(m,1H),7.93(s,1H),7.89(dd,J＝5.0,1.3Hz,1H),7.80(dd,J＝3.8,1.3Hz,1H),7.72(d,J＝8.2Hz,1H),7.54(td,J＝7.7,1.4Hz,1H),7.51–7.45(m,1H),7.40(d,J＝7.8Hz,1H),7.30–7.23(m,2H),7.20–7.13(m,1H),6.12(s,1H),4.44(t,J＝7.0Hz,1H),3.33(td,J＝3.3,1.7Hz,1H),3.19(dd,J＝13.5,7.7Hz,1H)。 13C NMR(101MHz,MeOD):δ180.54,171.36,153.45,144.45,139.78,137.43,136.64,136.64,136.52,136.52,135.86,134.72,132.92,131.91,130.79,130.59,130.46,130.46,129.76,129.76,128.56,128.24,127.79,124.83,124.66,122.00,118.03,89.80,59.41,39.74.HR-MS(m/z)(ESI):计算为C 30H 21F 3N 3O 5S 2[M-H]+:624.0880；实测值：624.0905。
实施例3’式(IV)的化合物的合成
将如上合成的1,4-萘醌-2-氨基酰化的取代苯胺类化合物与甲氧基胺盐酸盐以物质的量比为1:10加入装有25mL吡啶的圆底烧瓶中，控制温度在80℃下回流反应2h，减压旋转干燥，经洗脱剂为石油醚:乙酸乙酯(6:1)的硅胶柱纯化，旋转干燥得到的产品为黄绿色固体。所得产物具有如下具体结构式和参数表征。
R)-N-(3,5-二甲氧基苯基)-2-((4-(甲氧基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基丙酰胺(IVa)
Figure PCTCN2019095516-appb-000071
产率：70.6％。黄绿色固体。熔点169.1～169.5℃。 1H NMR(600MHz,DMSO-d 6)δ10.20(s,1H,NH),8.12(d,J＝7.7Hz,1H,C 6H 6),8.05(dd,J＝7.9,0.9Hz,1H,C 6H 6),7.74–7.65(m,1H,C 6H 6),7.62–7.55(m,1H,C 6H 6),7.32–7.25(m,4H,C 6H 6),7.22–7.17(m,1H,C 6H 6),6.80(d,J＝2.2Hz,2H,C 6H 6),6.49(d,J＝8.3Hz,1H,NH),6.37(s,1H,CH＝CH),6.25(t,J＝2.2Hz,1H,C 6H 6),4.37(dd,J＝13.8,7.8Hz,1H,CH),4.06(s,3H,CH 3),3.71(s,6H,),3.20(dd,J＝6.5,2.9Hz,2H,CH 2). 13C NMR(151MHz,DMSO-d 6))δ179.70,169.47,160.53,160.53,145.15,141.11,140.07,137.09,133.18,132.94,129.47,129.25,129.25,128.49,128.32,128.32,126.67,126.01,122.59,97.81,97.81,95.77,90.62,62.79,57.79,55.14,55.14,37.61. HR-MS(m/z)(ESI)：计算为C 28H 27N 3O 5[M+H] +:486.2023；实测值：486.2020。
实施例4靶向STAT3的体外抗肿瘤活性研究
4.1试剂的配制
PBS缓冲液的配制：取PBS粉末，先用800mL去离子水将其全部溶解，再加200mL去离子水定容到1L(10mmol/L)。在实验使用前将其高压灭菌，将灭菌后的PBS缓冲液在4℃条件下保存。
MTT染色剂的配制和使用：称取0.25g的MTT(methyl thiazolyl tetrazolium，又称为噻唑蓝)，用无酚红的培养基或磷酸缓冲液(PBS)溶解，定容到50mL。用滤膜滤除液体中的细菌，然后放入棕色瓶，在4℃下避光存放。
4.2 MTT法筛选
本实验为研究肟基萘醌类衍生物的体外抗肿瘤活性，选取人结肠癌细胞Hct-116、卵巢癌细胞SKOV3、肺小细胞癌细胞A549和肝癌细胞HepG2，利用MTT方法对本申请合成的化合物进行体外抗癌活性测试，其实验步骤如下。
(1)细胞的培养：人子宫颈癌细胞HeLa，置于含有10％胎牛血清和1％双抗的DMEM(Dulbecco’s modified eagle medium)培养基，并在37℃的温度、5％CO 2及95％空气的CO 2恒温培养箱中进行培养。待细胞贴壁长满后，进行传代，冻存。
(2)种板：在细胞的对数生长期间，用PBS洗2次后，再用0.25％胰蛋白酶消化，加入培养基终止消化，并小心吹打，得到单细胞悬浮液。经过计数，将细胞悬液用适量的10％培养基稀释并接种于96孔板中，每孔180μL，每孔细胞数为2×10 4至4×10 4。
(3)加样品：在种有不同癌细胞的96孔板中，每孔加入待测样品20μL(样品浓度为200μM)，最终样品浓度为20μM，进行初次筛选。经过筛选，挑选结果较好的样品进行不同浓度梯度再筛，计算IC 50，每一组都同为9个复孔。对照组每一孔放入20μL含10％DMSO的培养基，孵育48h。加样品作用48h后，每一孔放进10μL浓度为5mg/mL的MTT液体，在CO 2培养箱中再培养4h。
(4)测试：将孔内的培养基吸弃，并每孔加入100μL DMSO，在摇床上震荡10min，使甲瓒结晶完全溶解。然后用酶标仪(TECAN infinite M1000)(490nm)测定每孔的吸光度值。由如下公式来算癌细胞抑制率：抑制率(％)＝1-A 样品/A 对照×100％。根据相同样品的不同浓度对癌细胞生长抑制率制图，获剂量反应线，按照线性回归曲线算出IC 50，即细胞存活率下降50％时的样品浓度。
其中将阿霉素(DOX)作为阳性对照，具体测试结果见表1。
表1.本申请的化合物对于肿瘤细胞株Hct-116、SKOV-3和A549的细胞毒活性
Figure PCTCN2019095516-appb-000072
*(R)-2-(1,4-二氢-1,4-二氧代萘-3-基氨基)-3-苯基丙 酸，其为形成酰胺的1,4-萘醌取代的氨基酸，结构如下：
Figure PCTCN2019095516-appb-000073
**阿霉素(DOX)结构如下：
Figure PCTCN2019095516-appb-000074
从表1可知，本申请的化合物对所测试的肿瘤细胞株Hct-116、SKOV-3和A549表现出显著的体外抗肿瘤活性。末端芳基上的不同取代基对抗肿瘤活性可产生一定的影响。
在末端芳基上具有一个或多个吸电子基团可提高体外抗肿瘤活性。例如，具有一定空间电子特性来增强亲脂性的取代基，其穿透细胞膜的能力较强，可引起抗增殖效果的增强。
根据以上对肿瘤细胞系的活性筛选数据显示，化合物Iu对Hct-116、SKOV-3和A549的抗肿瘤活性分别为0.037±0.01μM、0.028±0.01μM、0.033±0.01μM和0.012±0.01μM，比阿霉素(分别为0.27±0.08μM、0.16±0.17μM、0.074±0.05μM和0.069±0.02μM)的好，尤其是针对人结肠癌细胞Hct-116和卵巢癌细胞SKOV3。
4.3 Western Blot蛋白印迹实验
4.3.1蛋白样品的制备
1.配制蛋白裂解液：10μl PMSF(100mM)中放入1mL裂解液，摇晃均匀置于冰上(PMSF需摇晃到没有出现晶体时才能够与裂解液混合)。
2.将加样品(样品浓度分别是0.1μM和0.5μM)后的培养液倒至15mL离心管中，离心时间为10min，以2000r/min的速度离心。
3.弃去上清液，加入3mL PBS轻轻吹打，反复洗涤两次，离心时间为10min，以2000r/min的速度，用枪吸干上清后置于冰上。
4.用预冷的缓冲液反复洗细胞3次后，将培养瓶置于冰上面。
5.将裂解液向离心管中加100μL，向培养瓶加400μL，一起放在冰上不断 晃动使细胞完全破裂，时间大约30min。
6.裂解完，用洁净的枪头将细胞刮下来，裂解液由稀变稠再变稀即可。在4℃条件下，把细胞碎片和所有的裂解液倒在15mL离心管中，离心时间为5min，速度为12000r/min。在冰上把上清液放到0.5mL EP管中，在-20℃下保存。
4.3.2 SDS-PAGE电泳
1.按说明把电泳仪装好，保证水平不漏胶。
2.把分离胶配好，加入TEMED混匀后灌胶，在该过程中尽可能不出现气泡。加到大约3/4时停止，并立刻用水封上层。
3.在水与胶中明显出现一线时，则胶凝完成。吸去上层的水，再用滤纸吸干水。放入提前制好的6％浓缩胶，立刻插梳子，在该过程中不应出现气泡。
4.待胶凝固，慢慢拔出梳子。将蛋白样品放到沸水中煮15分钟致使变性。放入5×SDS缓冲液使最终浓度为1×，加样为10μl，尽可能不溢出孔。对照组为Marker。
5.跑胶：在50V下进行半小时后(跑浓缩胶)，增加到100V跑1.5小时，将电泳置Marker全部分开，当指示剂移动到电泳槽底部时即可停止。
4.3.3转膜
1.准备：先将厚滤纸1张、中厚滤纸1张、薄滤纸2张浸泡于转膜缓冲液中，并将NC膜取出浸泡于二次水中。
2.在扁平盒中放入适量的转膜缓冲液，先放中厚滤纸，然后放薄滤纸，再放NC膜于转膜缓冲液中。将胶取下，切割胶使其大小与滤纸大小相当，水平放置于NC膜上，注意不能产生气泡。再往胶上放薄滤纸，最上层为厚滤纸，注意驱赶气泡。用镊子将最上层的厚滤纸与最下层的中厚滤纸分开，避免烧焦NC膜。轻轻将处理好的滤纸放入半干式的转膜仪中，转膜1.5小时。
3.转膜结束，用1×丽春红染液泡膜五分钟，二次水洗后出现条带，根据Marker的指示切胶。
4.3.4免疫反应
1.封闭。将切好的膜清洗并转移到含有脱脂牛奶的皿中，将皿在摇床上在室温条件下封闭摇晃1.5小时。然后用TBST清洗膜，即在摇床上摇动清洗。
2.按照1:500的比例稀释一抗，采用的倒扣的方法孵育一抗，采用摇床摇动孵育内参。将一抗孵育置于4℃过夜。
3.过夜孵育一抗后，用TBST洗涤一抗。反复洗涤5次，每次30分钟。按照1:1000的比例稀释二抗，也采用的倒扣的方法孵育二抗，采用摇床摇动孵育内参。将二抗孵育1.5小时。二抗孵育完成后，用TBST洗涮二抗。重复洗5次，每次10分钟。
4.3.5发光
1.将透明保鲜膜粘贴在发光夹子中，将膜放在两张保鲜膜中。向膜上滴加发 光液，在避光条件下观察荧光强度便于确定发光时间。
2.将胶片放入发光夹子中，盖上放光一定时间后，取出胶片马上放入显影液中。注意控制曝光时间，曝光到目的条带出现则可，以免曝光过度而太黑。曝光完毕后取出来放入定影液中数分钟，取出于清水中洗涤，晾干则可。免疫孵育后进行发光成像，结果见图1和图2。实验结果解析如下。
抑制STAT3磷酸化
STAT3中Tyr705的磷酸化导致STAT3的二聚、向核转移、与DNA特异性结合，并上调各种下游靶基因，如Bcl-xl、Bcl-2、Survivin、c-Myc、细胞周期蛋白D1(Bikash Debnath,Shili Xu,and Nouri Neamati.J.Med.Chem.2012,55,6645-6668)。如图1所示，经不同浓度的化合物Iu处理后，细胞中p-STAT3(Y705)的表达明显降低，而S727位点和总STAT3的表达没有发生改变(Y705和S727分别代表STAT3蛋白中编号分别为705和727的两个氨基酸残基)。这说明化合物Iu通过抑制Y705位点的磷酸化而抑制STAT3的激活，并且不是由于降低总STAT3的表达而抑制。
化合物Iu选择性抑制STAT3
STAT家族包含STAT1、STAT2、STAT3、STAT4、STAT5和STAT6等不同的亚型蛋白，在肿瘤细胞中STAT3常处于激活状态。为了验证化合物Iu对STAT3的选择性，通过蛋白质印记法检测了其他亚型蛋白的表达。图2结果表明化合物Iu对STAT1和STAT5的表达没有影响。为了进一步验证化合物Iu对STAT3的选择性，使用荧光素酶法检测了STAT1、STAT3、STAT4和STAT5的活性。
荧光素酶测定：将细胞接种于24孔板中，并与STAT3报告质粒4×M67 pTATA TK-Luc和Renilla萤光素酶对照报告质粒一起瞬时转染。然后将细胞与完全培养基一起用化合物Iu孵育24小时处理后，每个孔中用100μL细胞裂解缓冲液裂解细胞并离心收集上清液。取25μL上清液，使用双荧光素酶测定试剂盒，通过酶标仪测量STAT3的荧光素酶活性。结果见图3。
结果显示经化合物Iu处理细胞后荧光素酶活性的降低，而几乎没有观察到STAT1、STAT4和STAT5转录活性的变化。这些结果说明了化合物Iu能选择性抑制STAT3。选择性抑制STAT3，能够推断出对癌症的抑制活性。
抑制STAT3与DNA结合
采用酶联免疫吸附法(ELISA)进行检测。
酶联免疫吸附法(ELISA)：向每个孔中分别加入30μL完全结合缓冲液和20μL STAT3重组蛋白，将板在室温下温和搅拌1小时。使用20μL不含STAT3重组蛋 白的完全裂解缓冲液作为对照孔。孵育1小时后，用200μL 1X洗涤缓冲液洗涤3次，然后与100μL 1X STAT3抗体(1：1000稀释)孵育1小时，无需搅拌。将孔用200μL的1X洗涤缓冲液洗涤3次，并与100uL 1X的HRP缀合的抗体(1：1000稀释)孵育1小时，无需搅拌。最后，用200μL的1X洗涤缓冲液洗涤4次，然后使用显影溶液在室温条件下显色15分钟。加入终止溶液，立即使用酶标仪在450/655nm处读取吸光度值。
经不同浓度的化合物Iu处理细胞，并将样品进行ELISA测定。如图4所示，与不经化合物处理的样品相比，化合物Iu抑制STAT3-DNA结合活性呈浓度依赖性，因此证明化合物Iu是通过阻碍STAT3与DNA来抑制STAT3的激活。
4.4免疫荧光成像
将SKOV3细胞接种在无菌载玻片上并生长24小时。第二天将细胞用化合物Iu处理24小时。对于核易位实验，在无血清过夜后，用化合物Iu预处理SKOV3细胞2小时，然后再加入IL-6(interleukin-6，白细胞介素6)30分钟。处理后，用冷的磷酸盐缓冲盐水(PBS)缓冲液洗涤细胞，并在室温下用冷的甲醇固定15分钟。用预冷的PBS缓冲液洗涤两次后，在室温下用含有0.3％Triton的X-100透化细胞，用5％正常山羊血清的PBS缓冲液封闭至少1小时。然后用磷酸化的STAT3或STAT3多克隆兔抗体孵育细胞并在4℃下过夜。过夜孵育后，用含有0.1％Tween-20的PBS缓冲液洗涤细胞。在室温下将细胞与FITC荧光抗兔二抗孵育1小时。洗涤细胞后用DAPI进行，然后通过荧光显微镜进行成像分析。结果见图5，实验结果解析如下。
抑制STAT3的核转移
为了检测化合物Iu是否能够抑制STAT3转移到细胞核中，采用免疫荧光成像进行分析。如图5所示，在用IL-6处理SKOV3细胞中，STAT3被磷酸化并转位到细胞核中。然而，在用化合物Iu(0.5μM)处理的细胞中，大部分STAT3保留在细胞质中。因此，该结果表明化合物Iu可抑制STAT3与DNA结合以及磷酸化，而损害SKOV3细胞中STAT3的转录功能主要通过阻断其核转移。
4.5划痕实验和入侵实验
划痕实验
用含有10％FBS的DMEM培养基培养SKOV3细胞并将细胞接种在6孔板中培养。细胞长满后，使用10uL移液器尖端划痕，并用PBS洗涤1次以去除非贴壁细胞。将培养板置于倒置显微镜下观察并成像，结果作为不经处理的阴性对照。更换新鲜培养基，用化合物Iu处理细胞，将细胞置于培养箱中孵育24小时。孵育后在显微镜下观察记录细胞迁移情况。实验结果见图6，实验结果解析见下 文。
入侵实验
迁移测试在8.0μm聚碳酸酯膜的24孔入侵小室(transwell)中进行。首先，将600μL完全培养基或处理因子加入入侵小室下室。其次，将200μL含2×10 5/ml SKOV3细胞无FBS的DMEM悬浮液接种到入侵小室上室中，并将细胞迁移过滤器插入下部室中。在37℃下孵育24小时。然后，去除过滤器上侧的SKOV3细胞，用500μL 4％多聚甲醛固定迁移细胞20分钟，并用苏木精将固定的细胞染色3分钟。计数迁移细胞数量并在荧光显微镜下拍照。实验结果见图7，实验结果解析如下。
抑制SKOV3细胞迁移和入侵
肿瘤的转移和组织入侵是导致肿瘤患者死亡和治疗失败的主要因素之一，而STAT3的激活与肿瘤的增殖、分化、存活、转移和入侵有着密切联系。化合物Iu能阻断STAT3与DNA的结合从而抑制STAT3的转录活性，因此采用划痕实验和细胞小室染色分析检测化合物Iu能否抑制SKOV3细胞的迁移和入侵。如图6所示，化合物Iu抑制SKOV3细胞的迁移呈浓度依赖性。图7显示了相似的结果，随着浓度的增加，迁移的细胞数量逐渐减少。这些数据表明了化合物Iu能抑制SKOV3细胞的迁移和入侵。
抑制体内肝癌生长
从上海凌昌生物技术有限公司(中国)购买5周龄雄性BALB/c裸鼠，通过在裸鼠背部区域皮下注射100μL含1×10 7个人肝细胞癌HepG2细胞的无菌PBS诱导肿瘤形成，并将动物随机分成四组。在第15天当所有小鼠中肿瘤的体积达到100-150mm 3时，通过尾静脉注射的方式向第一组小鼠注射等体积的5％葡萄糖注射液媒介物作为对照。第二组每3天用阿霉素以5mg/kg体重的剂量治疗，持续3周。第3组和第4组用化合物Iu以10mg/kg或20mg/kg体重的剂量治疗，每三天一次，持续三周。药物处理后每隔一天记录肿瘤体积和体重。处理三周后处死所有小鼠，用电子数字卡尺测量肿瘤体积，通过测量长度(A)和宽度(B)计算体积(V＝AB 2/2。实验结果见图8A-8D，实验结果解析见下文。
如图8A所示，静脉内给予化合物Iu，剂量为10mg/kg或20mg/kg，显著抑制肿瘤生长，与阴性对照组相比导致肿瘤生长减少53.4％和59.0％(图8B)。同时，与多柔比星(DOX)相比，Iu还减小了肿瘤的体积(图8C)。值得注意的是，化合物8u在试验期间具有良好的耐受性，没有引起裸鼠体重的显著下降(图8D)。
实施例5 IDO1抑制活性研究
5.1 IDO1酶活性检测
根据试剂商提供的说明进行人IDO1酶活性测试(Tojo,S.；Kohno,T.；Tanaka,T.；Kamioka,S.；Ota,Y.；Ishii,T.；Kamimoto,K.；Asano,S.Med.Chem.Lett.2014,5,1119-1123)。将IDO抑制剂IDO5L作为阳性对照。将包含50μL反应预混液、10μL纯化的人IDO1、10μL待检测化合物、IDO1检测缓冲液的混合液(90μL)在室温条件下孵育10分钟，然后加入10μL含1mM L-色氨酸的基底溶液，使其终体积为100μL，37℃下避光孵育45分钟。加入50μL荧光显影试剂，在45℃下避光孵育3小时。室温冷却1小时后检测荧光强度(Ex/Em＝402/488nm)。根据公式[100-(A/B)100]/100计算抑制百分数，其中A为添加测试化合物的荧光强度，B为不添加测试化合物的荧光强度。实验结果见表2，实验结果解析见下文。
表2.本申请的化合物对于IDO1的抑制活性
Figure PCTCN2019095516-appb-000075
*(R)-2-(1,4-二氢-1,4-二氧代萘-3-基氨基)-3-苯基丙酸
**IDO5L为IDO抑制剂，其化学结构如下：
Figure PCTCN2019095516-appb-000076
hIDO1抑制活性
测试本申请所合成的化合物对人IDO1(hIDO1)(人吲哚胺2,3-双加氧酶1)的抑制活性，其中将IDO1抑制剂IDO5L作为阳性对照。如表2所示，测试结果表明本申请的肟基萘醌类衍生物对hIDO1具有良好的选择性抑制活性，IC 50以微摩尔每升的浓度计算，部分化合物的活性接近或稍好于阳性药物IDO5L(IC 50＝0.073μM)。
从表2中的数据可以初步推断出肟基萘醌类衍生物抑制hIDO1的构效关系。对肽末端芳基上位于间位的R 1取代基团的影响分析显示，与甲基或者甲氧基取代基团比较，卤素或者三氟甲基原子基团可在一定程度上增强衍生物对hIDO1的抑制活性。例如，当化合物Im末端芳基上间位甲基被三氟甲基取代时(化合物Ik)，hIDO1抑制活性显著增强，抑制活性相对于化合物Im提高了16.8倍。以氟原子或者氯原子取代芳基的对位分别得到化合物Il和Ih，产生了非常明显的抑制活性，具有富电子特性的取代基对hIDO1的抑制活性可起到一定的作用。当醌环上的肟被取代形成肟酯，活性有所降低(化合物IIIk-1、IIIq、IIIr)说明肟结构对于hIDO1抑制活性具有一定作用，原因可能是肟能与结合血红素的铁原子。
5.2紫外(UV)-可见光谱
紫外可见吸收光谱在Cary 1E UV-Visible的分光光度计上测量，扫描范围为200-700nm。为了保持各样品的温度在25℃下测定，将Haake F3水浴加热装置与分光光度计联用。测量使用1mL的样品磷酸盐缓冲溶液(PBS)水溶液，其中含有10μM纯化的IDO1和25μM的化合物Iu。实验结果见图9，结果解析如下。
紫外光谱检测化合物Iu与IDO1相互作用
由于化合物Iu对IDO1具有较好的抑制活性，因此将通过UV-可见光谱研究化合物Iu与IDO1的结合模式。IDO1具有血红素辅因子活性位点，其使得能够表征抑制剂与IDO间的相互作用。化合物与血红素铁结合能造成吸收光的改变，因此可以直接推断出与该结合位点结合。血红素为中心具有铁配位的卟啉环，根据其铁的氧化和配位状态，紫外可见光谱的最大吸收光波长大约为400nm。抑制剂与血红素的铁结合将改变其最大吸收光波长。血红素基团的吸收光谱对配体/底物结合后血红素周围极性的变化高度敏感，这改变了血红素的光谱特性。因此，由IDO1和配体相互作用引起的紫外-可见光谱的变化可用于评估化合物与IDO1 的结合。在本文中，采用紫外-可见光谱检测化合物Iu与IDO1相互作用。在没有化合物Iu的情况下，IDO1铁的吸收光谱在403nm出现Soret峰，这与之前的文献报道一致(Yi-Hui Peng,Shau-Hua Ueng,Chen-Tso Tseng,et al,J.Med.Chem.2016,59,282-293)。在化合物Iu存在的情况下，吸收光转移到413nm，这证明化合物Iu与IDO1结合并与血红素螯合(参见图9)。
5.3表面等离子体共振分析
使用表面等离子体共振(SPR)验证化合物Iu和IDO1蛋白之间的相互作用。将净化的人IDO1蛋白固定在传感器芯片CM5(GE)并且在25℃下在Biacore T200(GE，型号Biacore T200)仪器上进行分析。将HBS-EP缓冲液(0.01M HEPES、0.15M NaCl、3mM EDTA、0.05％v/v表面活性剂P20(pH7.4))作为运行缓冲液。比例缔合和解离速率常数被确定为结合亲和力(K D)。实验结果见图10，结果解析如下。
表面等离子共振检测化合物Iu和IDO1相互作用
为了进一步验证化合物Iu和IDO1之间的相互作用，使用GE Biacore T200光学生物传感器测定表面等离子共振(SPR)。如图10所示，在选定的浓度范围内，响应单位值(RU)与化合物的浓度成正比。经测试仪器自带软件计算，化合物Iu和IDO1之间的平衡解离常数(KD)是0.02μM，这充分证明了化合物Iu与靶标IDO1蛋白的结合亲和力。
抑制体内黑色素瘤的生长
通过皮下注射1×10 7培养的B16F10细胞在C57BL/6小鼠中建立黑素瘤模型。从第一天开始，将动物随机分成四组。随后，通过尾静脉注射给予5％右旋糖，D-1-MT(200mg/kg)，化合物Iu(100mg/kg)和化合物Iu(150mg/kg)，每两天一次，持续三周。药物处理后每隔一天记录肿瘤体积和体重。处理三周后处死所有小鼠，用电子数字卡尺测量肿瘤体积，通过测量长度(A)和宽度(B)计算体积(V＝AB 2/2)。实验结果见图11A-图11D，实验结果解析见下文。
为了研究抑制IDO1是否可以逆转体内免疫耐受，用化合物Iu或D-1-MT处理携带B16-F10的C57BL/6小鼠21天。21天后，化合物Iu抑制肿瘤的作用比阴性对照组和阳性对照更强(图11A)。比较化合物Iu处理3周后肿瘤的平均体积时，发现化合物Iu处理B16-F10模型小鼠引起显著的肿瘤生长抑制(图11C)，T/C比率为29.47％(150mk/kg)。具体地，化合物Iu减少了70.53％的肿瘤体积，而1-MT减少了57.21％(图11C)。用化合物Iu处理小鼠能显著地减少肿瘤总重量，而1-MT的抗肿瘤作用稍弱于化合物Iu(图11B)。每组老鼠的体重逐渐增加，表明化合物Iu的安全性较好(图11D)。

Claims (32)

  1. 式(I)的化合物：
    Figure PCTCN2019095516-appb-100001
    其中
    X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
    各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
    n为选自1、2、3、4或5的整数；
    各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
    m为选自1、2、3或4的整数；和
    R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
    或其药学上可接受的盐。
  2. 如权利要求1所述的化合物，其中所述C 6-C 10芳基为苯基。
  3. 如权利要求1所述的化合物，其中所述5元至10元杂芳基为吡啶基或噻吩基。
  4. 如权利要求1所述的化合物，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
  5. 如权利要求1所述的化合物，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
  6. 如权利要求1所述的化合物，其中所述各个R 1均为氢。
  7. 如权利要求1所述的化合物，其中所述各个R 2均为氢。
  8. 如权利要求1所述的化合物，其中所述R 3为氢。
  9. 选自以下的化合物：
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-(三氟甲基)苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3,N-二苯基-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲氧基苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲基苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氯苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-溴苯基)- 丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴-4-氟苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氟苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氯苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-溴苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲氧基苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲氧基苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟-4-溴苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二氟苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟-4-甲基苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2,5-二甲氧基苯基)-丙酰胺；
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯-4-甲基苯基)-丙酰胺；和
    (R)-2-((4-(羟基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲基-4-溴苯基)-丙酰胺；
    或其药学上可接受的盐。
  10. 式(II)的化合物：
    Figure PCTCN2019095516-appb-100002
    其中
    X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
    各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
    n为选自1、2、3、4或5的整数；
    各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
    m为选自1、2、3或4的整数；和
    R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
    或其药学上可接受的盐。
  11. 如权利要求10所述的化合物，其中所述C 6-C 10芳基为苯基。
  12. 如权利要求10所述的化合物，其中所述5元至10元杂芳基为吡啶基或噻吩基。
  13. 如权利要求10所述的化合物，其中所述C 6-C 10芳基任选地被选自以下的基团取代：卤素、羟基、巯基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基。
  14. 如权利要求10所述的化合物，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
  15. 如权利要求10所述的化合物，其中所述各个R 1、R 2均为氢，R 3为氢。
  16. 选自以下的化合物：
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-(三氟甲基)苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3,N-二苯基-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲氧基苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲基苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氯苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-溴苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴-4-氟苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-氟苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氯苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-溴苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲氧基苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(4-甲苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二甲氧基苯；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氟-4-溴苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3,5-二氟苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-氟-4-甲基苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2,5-二甲氧基苯基)-丙酰胺；
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-氯-4-甲基苯基)-丙酰胺；和
    (R)-2-((1,4-二氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(2-甲基-4-溴苯基)-丙酰胺；
    或其药学上可接受的盐。
  17. 式(III)的化合物：
    Figure PCTCN2019095516-appb-100003
    其中
    X为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、 任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
    各个R 1独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
    n为选自1、2、3、4或5的整数；
    各个R 2独立地选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；
    m为选自1、2、3或4的整数；
    R 3选自氢、卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、任选取代的卤代C 1-C 6烷氧基、亚磺酰基、磺酰基、S-磺酰氨基、N-磺酰氨基、O-氨基甲酰基、N-氨基甲酰基、O-硫代氨基甲酰基、N-硫代氨基甲酰基、C-酰氨基、和N-酰氨基；和
    R 4为任选地被一个或多个选自以下的基团取代的C 6-C 10芳基或5元至10元杂芳基：卤素、羟基、巯基、任选取代的烃硫基、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烯基、任选取代的C 1-C 6炔基、任选取代的C 1-C 6烷氧基、任选取代的氨基、任选取代的羟基C 1-C 6烷基、任选取代的卤代C 1-C 6烷基、和任选取代的卤代C 1-C 6烷氧基；
    或其药学上可接受的盐。
  18. 如权利要求17所述的化合物，其中所述C 6-C 10芳基为苯基，所述5元至10元杂芳基为吡啶基或噻吩基。
  19. 如权利要求17所述的化合物，其中所述C 6-C 10芳基为苯基，其任选地被选自以下的基团取代：卤素、任选取代的C 1-C 6烷基、任选取代的C 1-C 6烷氧基、和任选取代的卤代C 1-C 6烷基。
  20. 如权利要求17所述的化合物，其中所述R 1、R 2均为氢，R 3为氢。
  21. 如权利要求17所述的化合物，其中所述C 6-C 10芳基为苯基，所述5元至10元杂芳基为噻吩基。
  22. 选自以下的化合物：
    (R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-(三氟甲基)苯基)-丙酰胺；
    (R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-溴苯基)-丙酰胺；和
    (R)-2-((4-((((4-(叔丁基)苯基)磺酰基)氧)亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基-N-(3-甲氧基苯基)-丙酰胺；
    或其药学上可接受的盐。
  23. 制备权利要求10-16中任一项所述的化合物的方法，所述方法包括如下所示的步骤a)至e)：
    Figure PCTCN2019095516-appb-100004
    a)使邻苯二甲酸酐和L-苯丙氨酸反应，所述L-苯丙氨酸中的苯基任选地被1至n个R 1取代；
    b)使步骤a)得到的产物与C 2O 2Cl 2反应；
    c)使步骤b)得到的产物与X-NH 2反应；
    d)使步骤c)得到的产物与肼反应；
    e)使步骤d)得到的产物与1,4-萘醌反应，所述1,4-萘醌中的苯环任选地被1至m个R 2取代并且2位碳任选地被R 3取代，从而获得式(II)的化合物；
    其中X、R 1、n、R 2、m和R 3如权利要求10所述。
  24. 如权利要求23所述的方法，其中：
    步骤a)的反应条件为在55-85℃下、优选在70℃下在酸性溶液中反应10-14小时，优选12小时；
    步骤b)的反应条件为在0-15℃下、优选在0℃下在卤代烷烃溶液中反应10-14小时，优选12小时；
    步骤c)的反应条件为在0-15℃下、优选在0℃下在卤代烷烃溶液中反应20-40分钟，优选30分钟；
    步骤d)的反应条件为在室温下在醇类溶液中反应2.5至3.5小时；
    步骤e)的反应条件为在室温下在三乙胺、N,N-二甲基甲酰胺和水的混合溶液中反应18-24小时。
  25. 制备权利要求1-9中任一项所述的化合物的方法，所述方法包括如下所示的步骤f)：
    Figure PCTCN2019095516-appb-100005
    所述步骤f)为将式(II)的化合物与盐酸羟胺反应，优选地，所述步骤f)的反应条件为在70-80℃下在醇类溶液中反应10-14小时，优选12小时，从而获得式(I)的化合物，其中X、R 1、n、R 2、m和R 3为如权利要求1中所述。
  26. 制备权利要求17-22中任一项所述的化合物的方法，所述方法包括如下所示的步骤g)：
    Figure PCTCN2019095516-appb-100006
    所述步骤g)为将式(I)的化合物与ClSO 2R 4反应，优选地所述步骤g)的反应条件为在惰性气体中在0-15℃下、优选在0℃下在二氯甲烷中反应20-40分钟，优选30分钟，随后逐滴加入三乙胺并且反应10-20分钟；其中X、R 1、n、R 2、m、 R 3和R 4为如权利要求17中所述。
  27. (R)-N-(3,5-二甲氧基苯基)-2-((4-(甲氧基亚氨基)-1-氧代-1,4-二氢萘-2-基)氨基)-3-苯基丙酰胺或其药学上可接受的盐。
  28. 药物组合物，其包含如权利要求1-9、17-22或27中任一项所述的化合物或其药学上可接受的盐、或者权利要求25或26所述的方法制备的化合物或其药学上可接受的盐以及一种或多种药学上可接受的载体、稀释剂、赋形剂或其组合。
  29. 权利要求1-9、17-22或27中任一项所述的化合物或其药学上可接受的盐、权利要求25或26所述的方法制备的化合物或其药学上可接受的盐或者权利要求28所述的药物组合物在抑制STAT3和/或IDO1中的用途。
  30. 权利要求1-9、17-22或27中任一项所述的化合物或其药学上可接受的盐、权利要求25或26所述的方法制备的化合物或其药学上可接受的盐或者权利要求28所述的药物组合物在制备用于抑制STAT3和/或IDO1的药物中的用途。
  31. 权利要求1-9、17-22或27中任一项所述的化合物或其药学上可接受的盐、权利要求25或26所述的方法制备的化合物或其药学上可接受的盐或者权利要求28所述的药物组合物在制备用于治疗癌症的药物中的用途。
  32. 如权利要求31所述的用途，其中所述癌症选自结肠癌、卵巢癌、肝癌、膀胱癌、子宫颈癌和肺小细胞癌。
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