WO2015070766A1 - 一种n-苄基色胺酮衍生物及其制备方法和应用 - Google Patents

一种n-苄基色胺酮衍生物及其制备方法和应用 Download PDF

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WO2015070766A1
WO2015070766A1 PCT/CN2014/090947 CN2014090947W WO2015070766A1 WO 2015070766 A1 WO2015070766 A1 WO 2015070766A1 CN 2014090947 W CN2014090947 W CN 2014090947W WO 2015070766 A1 WO2015070766 A1 WO 2015070766A1
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group
compound
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benzyltryptamine
ido
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PCT/CN2014/090947
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English (en)
French (fr)
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杨青
匡春香
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复旦大学
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Priority to CA2930203A priority Critical patent/CA2930203A1/en
Priority to US15/036,384 priority patent/US10059712B2/en
Priority to AU2014350729A priority patent/AU2014350729B2/en
Priority to CN201480067750.9A priority patent/CN105829312B/zh
Priority to MX2016006217A priority patent/MX2016006217A/es
Priority to KR1020167015465A priority patent/KR20160103986A/ko
Priority to EP14861381.3A priority patent/EP3070089B1/en
Priority to JP2016552662A priority patent/JP6629218B2/ja
Priority to EA201690915A priority patent/EA201690915A1/ru
Publication of WO2015070766A1 publication Critical patent/WO2015070766A1/zh
Priority to ZA2016/03889A priority patent/ZA201603889B/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P25/00—Drugs for disorders of the nervous system
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    • A—HUMAN NECESSITIES
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    • A61P25/00—Drugs for disorders of the nervous system
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/12—Ophthalmic agents for cataracts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the invention belongs to the technical field of medicinal chemistry, and particularly relates to an N-benzyltryptamine derivative, a preparation method and application thereof.
  • Indoleamine-2,3-dioxygenase (IDO; MW 48,000; EC 1.13.11.42) is a heme-containing enzyme that is the first enzyme in the mammalian tryptophan metabolism pathway and is limited.
  • Speed enzyme catalyzes the oxidation of the essential amino acid, tryptophan, by the conversion of dioxygen to N-formyl kynurenine and is responsible for the cleaning of tryptophan in the human body.
  • IDO causes a microenvironment in which tryptophan is absent in the body, which in turn leads to the development of diseases closely related to tryptophan deficiency such as cancer, cataracts and neurological disorders. Therefore, the search for high-efficiency inhibitors based on IDO targets has become a research hotspot in drug development in recent years.
  • Interferon gamma is one of several potential IDO expression inducers. During sustained activation of high levels of interferon gamma stimulation, IDO reduces the availability of free serum tryptophan and thus also reduces serotonin production. These changes, combined with the accumulation of neuroactive kynurenine metabolites such as quinolinic acid (also induced by IDO), promote the development of neuropathy/psychiatric disorders and are a cause of multiple psychological disorders, as well as IDO activation and Tryptophan degradation is a trigger for the symptoms associated with chronic diseases.
  • neuroactive kynurenine metabolites such as quinolinic acid (also induced by IDO)
  • IDO activity also involves the development of age-related nuclear cataracts.
  • IDO is the first enzyme in the biosynthesis of UV filters in the lens and is the rate-limiting enzyme.
  • Ultraviolet filter compounds derived from tryptophan degradation (kynurenine and 3-hydroxykynurenine glucoside) modify the proteins present in the human lens. The amount of these UV filter compounds increases with age and causes the lens to become cloudy, leading to what is known as age-related nuclear cataract.
  • IDO expression also relates to suppression of immune responses by preventing local T-lymphocyte proliferation.
  • T-lymphocytes are very sensitive to the lack of tryptophan and in the absence of tryptophan, T-lymphocytes are arrested in the G1 phase of the cell cycle.
  • This T cell-mediated inhibition of immune response is a factor that causes many diseases, including autoimmune diseases, allogeneic rejection, neurodegenerative disorders, depression, bacterial or viral infections (eg, human immunodeficiency virus HIV). And cancer (Swanson et al. Am. J. Respir. Cell Mol. Biol. 2003 30, 311).
  • IDO inhibitors can be used for the inhibition of psychological disorders as well as for the treatment of other pathological features of IDO-mediated tryptophan metabolism pathways, including infections of viruses such as AIDS, bacterial infections such as Lyme disease and streptococcal infections. , neurodegenerative disorders (such as Alzheimer's disease, Huntington's disease, and Parkinson's disease), depression, cancer (including T-cell leukemia and colon cancer), eye diseases (such as cataracts and age-related yellowing), and Autoimmune diseases, etc.
  • a variety of in vitro assays can be used to screen (eg, high throughput screening), test reaction reference or IDO inhibitors of extracts obtained from natural sources. Activity, or determine its IDO inhibition kinetic constant.
  • IDO is closely related to a variety of disease pathogenesis and has been shown to be a target for major diseases such as cancer, Alzheimer's disease, depression, and cataracts.
  • IDO inhibitors have broad application prospects as drugs, but so far no suitable IDO inhibitors can be marketed as drugs. Therefore, it is of great theoretical significance and application value to find new and effective IDO inhibitors.
  • IDO inhibitor 1-MT (1-methyltryptophan) can enhance the sensitivity of tumor cells to immune stimulation of T cells in vitro; it can delay the growth of tumor cells and enhance chemotherapy in vivo in animal models. The anti-tumor effect of the drug, and it works for almost all spontaneous tumors. Unfortunately, most of the existing IDO inhibitors have low inhibitory efficacy. 1-MT is a commonly used IDO inhibitor in various in vitro and in vivo experiments, and its inhibition constant Ki is also only 34 ⁇ M.
  • an N-benzyltryptamine derivative having a structural formula represented by the following formula 1:
  • R 1 is hydrogen or fluorine
  • R 2 is -NR 3 R 4 ;
  • the R 3 and R 4 are each independently selected from the group consisting of H, a substituted or unsubstituted C1-C4 alkyl group, a substituted or unsubstituted C2-C4 alkenyl group, a substituted or unsubstituted C2- a C4 alkynyl group, a substituted or unsubstituted C3-C6 cycloalkyl group;
  • R 3 , R 4 together with an adjacent nitrogen atom constitute a substituted or unsubstituted 5-7 membered heterocyclic ring wherein the 5-6 membered saturated ring has 1-2 nitrogen atoms, and 0-2 a hetero atom selected from the group consisting of O, S;
  • substitution means that one or more hydrogen atoms on the group (preferably a hydrogen atom on a nitrogen atom) are substituted with a substituent selected from the group consisting of a C1-C4 alkyl group, a C1-C4 haloalkyl group, and an amine group protection.
  • Base preferably tert-butoxycarbonyl
  • halogen preferably one or more hydrogen atoms on the group (preferably a hydrogen atom on a nitrogen atom) are substituted with a substituent selected from the group consisting of a C1-C4 alkyl group, a C1-C4 haloalkyl group, and an amine group protection.
  • Base preferably tert-butoxycarbonyl
  • the nitrogen atom on the ring may optionally have an amine group. Protection base.
  • the amine protecting group is selected from the group consisting of:
  • the 5-7 membered heterocyclic ring is not a heteroaryl ring.
  • the 5-7 membered heterocyclic ring is a saturated heterocyclic ring, preferably a 5-6 membered saturated heterocyclic ring.
  • the 5-7 membered heterocyclic ring contains only one or two heteroatoms.
  • all of the heteroatoms in the 5-7 membered heterocyclic ring are N.
  • R 3 and R 4 are each independently selected from the group consisting of C 1 -C 4 alkyl; or R 3 , R 4 together with an adjacent nitrogen atom constitute a substituted or unsubstituted 5-6 member saturated.
  • R 3 and R 4 are not H at the same time.
  • R 2 is a cyclic imine.
  • R 2 is a substituted or unsubstituted group selected from the group consisting of:
  • substitution means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of C1-C4 alkyl, halogen.
  • R 1 is F.
  • the substituent R 2 is a cyclic imine or a dialkyl substituted amine
  • R 1 is hydrogen or fluorine
  • the derivative is a compound selected from the group consisting of:
  • R 1 and R 2 are as defined above.
  • the inert solvent is DMF, preferably anhydrous DMF.
  • the reaction in the step (d), is carried out at about 10 to 40 °C.
  • the reaction time is from 0.5 to 12 hours, preferably from 1 to 5 hours.
  • the molar ratio of the compound of the formula 5, R 2 H and triethylamine is 1: (1-2): (2-5).
  • the method further comprises the step (c) before the step (d):
  • reaction of step (c) is carried out in the presence of an initiator, preferably in the presence of AIBN.
  • the step (c) comprises: mixing with AIBN with NBS and then reacting with a compound of formula 4.
  • the inert solvent described in step (c) is dichloromethane, preferably anhydrous dichloromethane.
  • the reaction is carried out at 75 to 85 °C.
  • the reaction time is 15-17 hours.
  • the molar ratio of the compound of the formula 4, NBS and AIBN in the reaction is 1:1: (0.005 - 0.02).
  • the method further comprises the step (b) before the step (c):
  • the inert solvent is toluene, preferably anhydrous toluene.
  • the reaction in the step (b), is carried out at 100 to 120 °C.
  • the reaction time is from 3.5 to 4.5 hours.
  • the compound of the formula 3 in the step (b), the compound of the formula 3, The molar ratio to triethylamine is (0.2-0.5): (0.2-0.5): 1.
  • the method further comprises the step (a) before the step (b):
  • the inert solvent is dichloromethane, preferably anhydrous dichloromethane.
  • the reaction in the step (a), is carried out at room temperature (preferably 10 to 40 ° C).
  • the reaction time is from 1.5 to 2.5 hours.
  • the molar ratio of the compound of the formula 2 to the oxidizing agent is (0.5-1):1.
  • 5-methyl ruthenium was suspended in dry dichloromethane, and m-chloroperoxybenzoic acid was added in portions at 0 ° C, and stirred at room temperature for 1.5-2.5 hours. After TLC detection was completed, the reaction formed a white solid. , washed with ethyl acetate three times to obtain 5-methyl isotonic anhydride; wherein, the molar ratio of 5-methyl ruthenium and m-chloroperoxybenzoic acid is: (0.5-1): 1;
  • the 2-methyltryptamine obtained in the step (2) was dissolved in dry dichloromethane at 75-85 ° C under nitrogen atmosphere, and then a mixture of NBS and AIBN was added in portions.
  • the reaction solution was heated at 75 ° C for 15-17 hours and the reaction was complete by TLC.
  • the reaction solution is cooled to room temperature, washed with brine, dried over anhydrous sodium sulfate and concentrated to obtain a yellow product; wherein the molar ratio of 2-methyltryptamine to NBS and AIBN is 1:1: (0.005-0.02);
  • the bromo 2-methyltryptamine, the fatty amine and the triethylamine were stirred in a dry DMF at room temperature for 1.5-2.5 hours. After the reaction was completed by TLC, 50 ml of water was added, and then extracted three times with 10 ml of dichloromethane, and then combined. The methylene chloride phase was washed three times with water and dried over anhydrous sodium sulfate. The yellow solid obtained by concentration is separated with silica gel to give a yellow N-benzyltryptamine derivative; wherein the molar ratio of bromo 2-methyltryptamine and fatty amine to triethylamine is 1: (1-2) :(2-5).
  • a N-benzyltryptamine derivative according to the first or second aspect of the present invention which is useful for the preparation of a disease preventing and/or treating an IDO-mediated disorder of tryptophan metabolism
  • the application of pharmacological characteristics of the disease in medicine is provided.
  • a compound of formula 1 or a pharmaceutically acceptable salt thereof for:
  • the disease associated with a tryptophan metabolism disorder is selected from the group consisting of a tumor, depression, anxiety, AIDS, autoimmune disease, psychological disorder, Lyme disease infection, streptococcal infection, neurodegenerative Conditions (eg Alzheimer's disease, Huntington's disease and Parkinson's disease), depression, cancer (including T-cell leukemia and colon cancer), eye diseases (eg cataracts and age-related yellowing), or autoimmune disease.
  • a pharmaceutical composition comprising (i) a compound of formula 1 according to the first or second aspect of the invention, or a pharmaceutically acceptable salt thereof; (ii) a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is for the treatment of a disorder associated with a disorder of tryptophan metabolism.
  • the disease associated with a tryptophan metabolism disorder is selected from the group consisting of a tumor, depression, Anxiety disorders, AIDS, autoimmune diseases, psychological disorders, Lyme disease infections, streptococcal infections, neurodegenerative disorders (eg Alzheimer's disease, Huntington's disease and Parkinson's disease), depression, cancer (including T-cell leukemia) And colon cancer), eye diseases (such as cataracts and age-related yellowing), or autoimmune diseases.
  • an IDO inhibitor comprising a compound of formula 1 according to the first or second aspect of the invention, or a pharmaceutically acceptable salt thereof.
  • a ninth aspect of the invention provides a method of non-therapeutic inhibition of IDO activity in vitro, the method comprising the steps of: inhibiting an effective amount of a compound of formula 1 according to the first or second aspect of the invention, or Its pharmaceutically acceptable salt is in contact with the subject to be inhibited.
  • a method of preparing a pharmaceutical composition for treating a disease associated with a disorder of tryptophan metabolism comprising: treating a therapeutically effective amount of the formula according to the first or second aspect of the invention A compound or a pharmaceutically acceptable salt thereof is mixed with a pharmaceutically acceptable carrier to form a pharmaceutical composition.
  • a method for treating a disease associated with a disorder of tryptophan metabolism comprising: administering to a subject a therapeutically effective amount of Formula 1 according to the first or second aspect of the present invention a compound, or a pharmaceutically acceptable salt thereof.
  • the disease associated with a tryptophan metabolism disorder is selected from the group consisting of a tumor, depression, anxiety, AIDS, autoimmune disease, psychological disorder, Lyme disease infection, streptococcal infection, neurodegenerative Conditions (eg Alzheimer's disease, Huntington's disease and Parkinson's disease), depression, cancer (including T-cell leukemia and colon cancer), eye diseases (eg cataracts and age-related yellowing), or autoimmune disease.
  • the present inventors After long-term and intensive research, the present inventors have unexpectedly found that a class of N-benzyltryptamine derivatives having a structure as shown in Formula 1 have relatively excellent IDO inhibitory activity, and the compounds are somewhat related to some and Compared with similar N-benzyltryptamine derivatives, the IDO inhibitory activity (IC 50 ) is extremely significant (IC 50 value can be reduced by 1-3 orders of magnitude), so it has a good application prospect. Based on the above findings, the inventors completed the present invention.
  • C1-C4 alkyl refers to a straight or branched alkyl group having from 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, Tert-butyl, or a similar group.
  • C2-C4 alkenyl refers to a straight or branched alkenyl group having 2 to 4 carbon atoms, such as a vinyl group, a propenyl group, a butenyl group, or the like.
  • C2-C4 alkynyl refers to a straight or branched alkynyl group having 2 to 4 carbon atoms, such as an ethynyl group, a propynyl group, a butynyl group, or the like.
  • 5-7 membered heterocyclic ring refers to a monocyclic ring having the meaning of 5 to 7 members, said ring having no fully conjugated pi-electron system.
  • the ring may optionally have from 1 to 3 heteroatoms, and the hetero atom includes O or N.
  • saturated rings include piperazinyl, morphinolinyl and the like.
  • substituted means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of C1-C4 alkyl, C1-C4 haloalkyl, and an amine protecting group (eg, tert-butyl). Oxycarbonyl), halogen.
  • halogen refers to F, Cl, Br and I.
  • the tryptamine is an quinazoline alkaloid whose chemical name is ⁇ [2,1-b]quinazoline-6,12-dione. Tryptamine is a yellow needle crystal which is mainly found in blue plants such as horse blue, indigo and indigo. Alternatively, it may be extracted from a fermentation broth of microorganisms.
  • tryptamine can be extracted from the metabolites of blue-producing plants and microorganisms, the separation process is long and the extraction rate is low, which is difficult to meet the needs of research and clinical use. Only through the exploration of short-time, high-yield, easy-to-obtain synthetic routes can provide more resources for the application of tryptamine, making it possible to further develop and apply.
  • the present invention structurally reforms tryptamine to improve the solubility and pharmacological activity of tryptamine, with the aim of obtaining an active compound having application value.
  • the research and pharmacological tests of the present invention show that the N-benzyltryptamine derivative formed by introducing a water-soluble group into the tryptamine molecule can be used as a more efficient IDO inhibitor, which has antibacterial, anti-inflammatory, anti-tumor, etc.
  • a variety of pharmacological activities has broad application prospects.
  • the synthesis method of the present invention has the advantages of simple operation, mild conditions, high yield, and the like, and is easier to industrially produce than the existing synthesis method of the tryptophan derivative.
  • N-benzyltryptamine derivative of the present invention has a structural formula represented by the following formula 1:
  • R 1 is hydrogen or fluorine
  • R 2 is -NR 3 R 4 ;
  • the R 3 and R 4 are each independently selected from the group consisting of a C1-C4 alkyl group; or R 3 and R 4 together with an adjacent nitrogen atom constitute a substituted or unsubstituted 5-6 membered saturated ring, wherein The 5-6 membered saturated ring has at least one nitrogen atom, and optionally 1-2 heteroatoms selected from the group consisting of O, N;
  • the substitution means that one or more hydrogen atoms (preferably a hydrogen atom on a nitrogen atom) on the group are substituted with a substituent selected from the group consisting of a C1-C4 alkyl group, a t-butoxycarbonyl group, and a halogen.
  • said R 2 has 1 nitrogen atom and 1 hetero atom selected from the group consisting of O and N.
  • said R 2 is a substituted or unsubstituted group selected from the group consisting of: among them, Indicates a connection site;
  • substitution means that one or more hydrogen atoms on the group are substituted with a substituent selected from the group consisting of C1-C4 alkyl, halogen.
  • the compound of the present invention its tautomeric form, its structural analog or a pharmaceutically acceptable salt thereof, and a composition containing at least one of the compound, a structural analog thereof or a pharmaceutically acceptable salt thereof may be used.
  • diseases include, but are not limited to, tumors, cancer, eye diseases, autoimmune diseases, psychological disorders, depression, and anxiety disorders.
  • Such uses include in vivo and in vitro applications, as well as in the preparation of pharmaceuticals, IDO inhibitors, and pharmaceutical compositions.
  • the inventors have found that, in a particularly preferred embodiment, when R 1 is F, the compound has the best IDO inhibitory activity.
  • tryptamine In recent years, pharmaceutical chemists have been working on the synthesis of tryptamine and its derivatives.
  • the main method of synthesizing tryptamine is the reaction of hydrazine with isatoic anhydride.
  • the method has simple yield and mild reaction conditions.
  • functionalized groups can be introduced on both the ruthenium and isatoic anhydride precursors to synthesize various functionalized tryptamines.
  • the most important method for synthesizing hydrazine is to use hydration of trichloroacetaldehyde, hydroxylamine and aniline in an aqueous solution of hydrochloric acid to form an anthraquinone compound, and then ring-closed to obtain hydrazine under concentrated sulfuric acid.
  • This method is very suitable for synthesis of The yield of halogen and alkyl tryptamine is also high.
  • this method is difficult to synthesize the active group-containing tryptamine because these groups are prone to various side reactions during the synthesis of hydrazine.
  • the inventors succeeded in obtaining a synthetic method for preparing tryptone having a reactive group. Specifically, the method includes the steps of:
  • the method further includes the steps of:
  • the bromination reaction is carried out in the presence of an initiator such as AIBN.
  • an initiator such as AIBN.
  • a preferred embodiment comprises: mixing with AIBN with NBS and then reacting with the compound of formula 4 should.
  • the method further includes the steps of:
  • the method further includes the steps of:
  • reaction time, reaction temperature and the like of the above respective steps can be adjusted according to actual conditions, for example, the end point of the reaction is determined by a method commonly used in the art (e.g., TLC method).
  • the solvent of each step is not particularly limited, and an inert solvent which is conventionally used in the art and which does not react with the reactant can be used. It should be understood that, after the above-described reaction route is disclosed in the present application, the specific conditions of each step can be determined by those skilled in the art in combination with the knowledge in the prior art.
  • an optimal synthetic route is as follows:
  • 5-methyl ruthenium was suspended in dry dichloromethane, and m-chloroperoxybenzoic acid was added in portions at 0 ° C, and stirred at room temperature for 1.5-2.5 hours. After TLC detection was completed, the reaction formed a white solid. The mixture was washed three times with ethyl acetate to give 5-methylindole anhydride; wherein the molar ratio of 5-methylisatin to m-chloroperoxybenzoic acid was from 0.5 to 1:1.
  • the 2-methyltryptamine obtained in the step (2) was dissolved in dry dichloromethane at 75-85 ° C under nitrogen atmosphere, and then a mixture of NBS and AIBN was added in portions. The reaction solution was heated at 75 ° C for 15-17 hours and the reaction was complete by TLC. The reaction solution was cooled to room temperature, washed with brine, dried over anhydrous sodium sulfate, and then evaporated and evaporated.
  • the bromo 2-methyltryptamine, the fatty amine and the triethylamine were stirred in a dry DMF at room temperature for 1.5-2.5 hours. After the reaction was completed by TLC, 50 ml of water was added, and then extracted three times with 10 ml of dichloromethane, and then combined. The methylene chloride phase was washed three times with water and dried over anhydrous sodium sulfate. The yellow solid obtained by concentration was separated with silica gel to give a yellow N-benzyltryptamine derivative. Wherein, the molar ratio of bromo 2-methyltryptamine and fatty amine to triethylamine is 1:1-2:2-5.
  • the N-benzyltryptamine derivative of the present invention has excellent IDO inhibitory activity, it can be used for the preparation of a medicament for preventing and/or treating a disease having a pathological characteristic of IDO-mediated tryptophan metabolism disorder (e.g., Tumors, etc.).
  • a disease having a pathological characteristic of IDO-mediated tryptophan metabolism disorder e.g., Tumors, etc.
  • the tryptophan metabolism disorder disease may be any disease known or unknown in the art having pathological features of IDO-mediated disorder of tryptophan metabolism, preferably the disease associated with the tryptophan metabolism disorder is selected from the group consisting of Lower group: tumor, depression, anxiety, AIDS, autoimmune disease, psychological disorder, Lyme disease infection, streptococcal infection, neurodegenerative disorders (eg Alzheimer's disease, Huntington's disease and Parkinson's disease), depression Disease, cancer (including T-cell leukemia and colon cancer), eye diseases (such as cataracts and age-related yellowing), or autoimmune diseases.
  • Lower group tumor, depression, anxiety, AIDS, autoimmune disease, psychological disorder, Lyme disease infection, streptococcal infection, neurodegenerative disorders (eg Alzheimer's disease, Huntington's disease and Parkinson's disease), depression Disease, cancer (including T-cell leukemia and colon cancer), eye diseases (such as cataracts and age-related yellowing), or autoimmune diseases.
  • Lower group tumor, depression, anxiety, AIDS, autoimmune disease,
  • the present invention provides a novel class of IDO inhibitors which have surprisingly improved activity compared to structurally similar compounds.
  • the compound has both IDO inhibitory activity and can also be used as a further modified pharmaceutical intermediate, and has potential application value for developing a new cancer therapeutic drug.
  • the present invention also provides a method for synthesizing a N-benzyltryptamine derivative, which has the advantages of simple operation, mild reaction conditions, solvent saving, pollution reduction, and the like, and is convenient for industrial production.
  • Step b Et 3 N, toluene, reflux, 75%;
  • Step c NBS, AIBN, CH 2 Cl 2 , reflux;
  • Step (1) - Step (3) is the same as in Example 1.
  • Step (1) - Step (3) is the same as in Example 1.
  • Step (1) - Step (3) is the same as in Example 1.
  • Step (1) - Step (3) is the same as in Example 1.
  • Step (4) Synthesis of 8-fluoro-2-((morpholine-1-yl)methyl)indolo[2,1-b)quinazoline-6,12-dione (1e)
  • Step (1) is the same as Embodiment 1,
  • the construction of the plasmid containing the human IDO gene, expression, extraction and purification in E. coli were carried out according to the method reported by Littlejohn et al. (Takikawa O, Kuroiwa T, Yamazaki F, et al. J. Biol. Chem. 1988, 263, 2041-2048).
  • the inhibitory activity of each compound on IDO was measured by the method described in the literature. 50 mM potassium phosphate buffer (pH 6.5), 40 mM vitamin C, 400 ⁇ g/ml catalase, 20 ⁇ M methylene blue and IDO enzyme were mixed in a 96-well plate. The substrate L-tryptophan and the sample to be tested were added to the above mixture.
  • the reaction was carried out at 37 ° C for 60 minutes, and the reaction was terminated by the addition of 30% (w/v) trichloroacetic acid.
  • the 96-well plate was heated at 65 ° C for 15 minutes to complete the conversion from formyl kynurenine to kynurenine, followed by a rotation of 6000 g for 5 minutes. Remove 100 ⁇ l of supernatant from each well and transfer to a new 96-well plate. Add 2% (w/v) p-(dimethylamino)benzaldehyde in acetic acid solution. Yellow color produced by reaction of kynurenine with it. It can be observed at 490 nm using a microplate reader, and the experimental results are shown in Table 1.
  • Example 8 is a reversible inhibitor
  • Reaction conditions In a 500 ⁇ l reaction system, first add 50 mM potassium phosphate buffer (pH 6.5), 40 mM vitamin C, 400 ⁇ g/ml catalase, 20 ⁇ M methylene blue, 300 mM substrate L-tryptophan or simultaneously add 100 mM inhibition. The mixture was incubated at 37 ° C for 5 minutes, and different volumes of IDO enzyme were added to the mixture. The reaction was carried out at 37 ° C for 30 minutes, and 30 ⁇ l (w/v) trichloroacetic acid 200 ⁇ l was added to terminate the reaction.
  • the reaction was terminated by adding 30 ⁇ l (w/v) trichloroacetic acid 200 ⁇ l, and the reaction system was heated in a 65 ° C water bath for 15 minutes to complete the conversion from formyl kynurenine to kynurenine, and then centrifuged at 12,000 rpm for 10 minutes. The supernatant was mixed with an equal volume of 2% (w/v) p-dimethylaminobenzaldehyde in acetic acid, and the 490 nm wavelength reading was detected by a microplate reader.
  • the inhibitor type of the compound was determined by Dixon's mapping method (1/v to [I]), and the Ki value of the inhibitor was obtained by plotting S/v to [I]. The experimental results are shown in Table 1.
  • the reaction was carried out at 37 ° C for 30 minutes, 30 ⁇ l (w / v) trichloroacetic acid 200 ⁇ l was added to terminate the reaction, and the reaction system was heated at 65 ° C for 15 minutes to complete the reaction from formyl kynurenine to kynurenine. Transformation, then spinning at 12000 rpm for 10 minutes, taking 200 ⁇ l of the supernatant and mixing with an equal volume of 2% (w/v) p-dimethylaminobenzaldehyde in acetic acid solution.
  • the yellow color produced by the reaction of kynurenine can be used with a microplate reader. detection at 490nm, the results obtained using calculation software IC 50, IC 50 value of an inhibitor to obtain experimental results as shown in table 1.
  • Plasmid pcDNA3.1-hIDO was transiently transfected into HEK 293 cells using liposome Lipofectamin 2000.
  • HEK293 cell culture medium was high glucose DMEM containing 50 U/mL penicillin, 50 U/mL streptomycin, 10% FBS, 37 ° C, 5% CO 2 culture.
  • the drug to be tested was added.
  • the supernatant was taken to another 96-well plate, 10 ⁇ L of 30% (w/v) trichloroacetic acid was added, and the mixture was heated at 65 ° C for 15 min to complete the nail.
  • the IDO inhibitory activity of the compounds prepared in Examples 1-6 was determined by the methods of the above Examples 7-11, and the IDO inhibitor 1-methyltryptophan (1-MT, which is commonly used in current and in vitro experiments) was used. As a control, the measurement results are shown in Table 1.
  • mice Two mice were sacrificed by cervical dislocation, and the spleen was taken out in a clean bench and placed in a 6 cm plate containing RP1640 medium for use.
  • Treated LLC cells 2 ⁇ 10 4 / well (stimulated cells), spleen lymphocytes 10 5 / well (reactive cells), add 96-well plates, add RP1640 (10% FBS), fill up to 200 ⁇ L ;
  • the administration group was added with 50 ⁇ M IDO inhibitor, and cultured in an incubator at 37 ° C, 95% humidity, 5% CO 2 , and cultured for 72 hours;
  • the WST-1 kit measures the proliferation of T cells, and the plate reader measures the absorbance at a wavelength of 450 nm;
  • T lymphocyte proliferation rate (%) [administration hole (T lymphocyte + LLC cell + IDO inhibitor) OD value
  • Example 1 Example 2
  • Example 3 Example 4
  • Example 5 T cell proliferation rate 4.88% 38.48 40.64 41.24 50.27 39.40 13.84
  • Example 138 Fluoro-2-((4-methylpiperazin-1-yl)methyl)indolo[2,1-b]quinazoline-6,12-dione (Example 4) Tumor effect
  • mice were randomly divided into 5 groups, 10 in each group.
  • the control group was given an equal amount of 0.5% sodium carboxymethylcellulose and administered intragastrically; the 1-methyl tryptophan (1-MT) high-dose group was 150 mg/kg, and the low-dose group was 75 mg/kg.
  • Tumor long diameter (a) and short diameter (b) were measured from the day of tumor attachment and were measured every other day.
  • Tumor volume ab ⁇ 2 /2.
  • the mice were sacrificed by the neck of the next day after stopping the drug, and the tumor was weighed and stored in liquid nitrogen.
  • mice were given a one-week prophylactic administration and continued to be administered after inoculation, and given 8-fluoro-2-((4-methylpiperazin-1-yl)methyl)anthracene.
  • the growth of Lewis lung cancer tumors was significantly inhibited in mice, and there was no obvious tumor formation on the 20th day after the start of high-dose administration, while the control group Tumors were found on day 11 after administration, and tumors occurred on the 13th day after administration in the 1-MT group.
  • mice in the 8-fluoro-2-((4-methylpiperazin-1-yl)methyl)indolo[2,1-b]quinazoline-6,12-dione group were normal in weight, The fur is shiny and the reaction is flexible.
  • mice were randomly divided into 4 groups, 10 in each group. When the tumor reached a diameter of 5 mm, it was administered for the first time.
  • the control group was given an equal amount of 0.5% CMC (carboxymethylcellulose sodium) and administered by intragastric administration; in the 1-MT group, the dose was 200 mg/kg, administered by intragastric administration; 8-fluoro-2-(4 -methylpiperazin-1-yl)methyl)indolo[2,1-b]quinazoline-6,12-dione group, administered in an amount of 200 mg/kg, administered intragastrically; paclitaxel group , the dosage is 100mg/kg, tail vein injection, once a week; 8-fluoro-2-((4-methylpiperazin-1-yl)methyl)indolo[2,1-b]quina Oxazoline-6,12-dione + paclitaxel group, 8-fluoro-2-
  • the mice were sacrificed by the neck of the next day after stopping the drug, and the tumor was weighed and stored in liquid nitrogen.
  • mice were treated with 200 mg/kg of 8-fluoro-2-((4-methylpiperazin-1-yl)methyl)indole[2,1-b]quinine
  • 8-fluoro-2-((4-methyl) The piperazine-1-yl)methyl)indolo[2,1-b]quinazoline-6,12-dione group was more effective in inhibiting tumor growth than the 1-MT group.
  • mice were found to have thin body shape, loose fur and less movement. And 8-fluoro-2-((4-methylpiperazin-1-yl)methyl)indolo[2,1-b]quinazoline-6,12-dione alone can not cause tumors. Regressed, but can inhibit tumor growth to a greater extent, and the mice in this group have normal body weight, shiny coat and flexible reaction.
  • Step (1) is the same as in Example 1.
  • the activity of the compound obtained in the comparative example was determined by the method as described in Examples 9-11, and the IC50 of the IDO inhibitory activity was 997.25 uM in vitro, and Ki and in vivo IC 50 were not detected.
  • the compound of the present application has an abnormally excellent IDO inhibitory activity as compared with the IDO inhibitor of the prior art and the compound having a similar structure.
  • the cells in vitro IC 50 values of the compounds of the present application have reduced 1-3 orders of magnitude, indicating that the compounds of the present application has very excellent IDO Inhibition activity.
  • the compounds of the present application have a significant improvement in inhibitory activity compared to some compounds which are very similar in structure (e.g., comparative compounds). Even with some IDO inhibitors having the same active groups in the structure, the inhibitory activity is also considerably improved.
  • the difference in biological activity between the compounds of the present application and their analogous structures is a rare exception compared to the correlation of structure and activity of compounds conventional in the art. This indicates that the compounds of the present application are a fairly unique class of structures whose biological activity is surprising.

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Abstract

本发明涉及一种N-苄基色胺酮衍生物及其制备方法和用途。本发明一种N-苄基色胺酮类衍生物，其特征在于，所述衍生物具有式1所示的结构通式；其中，各基团的定义如说明书中所述。本发明化合物的制备方法操作简单、条件温和、收率高，易于工业化生产等优点。本发明所述的N-苄基色胺酮衍生物具有优良的吲哚胺-2,3-双加氧酶（IDO）抑制活性，可应用于治疗具有IDO介导的色氨酸代谢的病理学特征的疾病。

Description

一种N-苄基色胺酮衍生物及其制备方法和应用 技术领域
本发明属于药物化学技术领域，具体涉及一种N-苄基色胺酮衍生物及其制备方法和应用。
背景技术
吲哚胺-2,3-双加氧酶(简称IDO；MW 48,000；EC 1.13.11.42)是含有血红素的酶，该酶是哺乳动物色氨酸代谢途径中的第一个酶并且是限速酶。IDO催化必需氨基酸—色氨酸通过双氧转化为N-甲酰犬尿氨酸的氧化反应，并且负责清理人体中的色氨酸。IDO通过降解色氨酸，造成体内色氨酸缺失的微环境，进而导致了癌症、白内障、神经紊乱等多种与色氨酸缺失密切相关的疾病的发生。因此，寻找基于IDO靶点的高效抑制剂已成为近年来药物开发的研究热点。
干扰素γ是若干潜在的IDO表达诱导剂中的一种。在高水平干扰素γ刺激的持续活化期间，IDO降低了游离血清色氨酸的利用度，因而也减少了5-羟色胺的产生。这些变化与诸如喹啉酸的具有神经活性的犬尿氨酸代谢物的蓄积(也由IDO诱导)相结合，促进神经病/精神病病症的发生并且是多种心理障碍的诱因，也是具有IDO活化和色氨酸降解特征的慢性病的相关症状的诱因。
IDO活性还涉及与年龄相关的核性白内障的发生。IDO是晶状体中紫外线滤器生物合成中的第一个酶，并且是限速酶。来自色氨酸降解的紫外线滤器化合物(犬尿氨酸和3-羟基犬尿氨酸葡萄苷)修饰存在于人晶状体中的蛋白质。这些紫外线滤器化合物的量随着年龄增长而增加,并且会导致晶状体逐渐浑浊，进而导致被称为与年龄相关的核性白内障。
IDO表达还涉及通过阻止局部T-淋巴细胞增殖而进行的免疫应答抑制。T-淋巴细胞对色氨酸的缺乏非常敏感并且在色氨酸缺失条件下，T-淋巴细胞停滞在细胞周期的G1期。这种T细胞介导的免疫应答抑制是导致许多疾病的因素，所述疾病包括自身免疫性疾病、异体排斥反应、神经退行性病症、抑郁症、细菌或病毒感染(例如人免疫缺陷病毒HIV)和癌症(Swanson et al.Am.J.Respir.Cell Mol.Biol.200330,311)。
已经发现，大多数人类肿瘤组成性地表达IDO。来自预先免疫小鼠的小鼠肿瘤细胞已经显示可以通过表达IDO而保护其不受排斥反应，通过1-MT的给药消除了上述效应。然后通过IDO抑制剂的伴行给药改善了癌症治疗的有效性。
IDO抑制剂可以用于心理障碍的抑制以及治疗其他具有IDO介导的色氨酸代谢途径的病理学特征的疾病，这些疾病包括诸如AIDS等病毒的感染、莱姆病和链球菌感染等细菌感染、神经退行性病症(例如阿尔茨海默病、亨廷顿病和帕金森病)、抑郁症、癌症(包括T细胞白血病和结肠癌)、眼部疾病(例如白内障和与年龄相关的黄化)以及自身免疫性疾病等。可以使用多种体外分析(Takikawa,et al.J.Biol.Chem.1998,263,2041)来筛选(例如高通量筛选)、测试反应参照物或从天然来源得到的提取物的IDO抑制剂活性，或确定其IDO抑制动力学常数。
IDO与多种疾病发病机制密切相关，已被证实它是癌症、阿尔茨海默病、抑郁症、白内障等重大疾病的靶标。IDO抑制剂作为药物具有广阔的应用前景，但是迄今未有合适的IDO抑制剂可作为药物上市，因此，寻找新型高效的IDO抑制剂具有重要的理论意义和应用价值。
现有研究表明，IDO抑制剂1-MT(1-甲基色氨酸)在体外能增强肿瘤细胞对T细胞的免疫刺激的敏感性；在动物模型中能体内延缓肿瘤细胞的生长并增强化疗药物的抗肿瘤效果，而且对几乎所有的自发性肿瘤都起作用。但遗憾的是，现有的IDO抑制剂大都抑制效力低下，1-MT作为各种体内外实验中常用的IDO抑制剂，其抑制常数Ki也仅为34μM。
因此，本领域迫切需要开发新型高效的IDO抑制剂。
发明内容
本发明的目的在于提供一种N-苄基色胺酮类衍生物及其制备方法和应用。
本发明的第一方面，提供了一种N-苄基色胺酮类衍生物，所述衍生物具有如下式1所示的结构通式：
Figure PCTCN2014090947-appb-000001
式中
R1为氢或氟；
R2为-NR3R4；
所述的R3、R4各自独立地选自下组：H、取代的或未取代的C1-C4烷基、取代的或未取代的C2-C4烯基、取代的或未取代的C2-C4炔基、取代的或未取代的C3-C6环烷基；
或R3、R4与相邻的氮原子共同构成取代或未取代的5-7元杂环，其中，所述的5-6元饱和环上具有1-2个氮原子，和0-2个选自下组的杂原子：O、S；
所述的取代指基团上的一个或多个氢原子(优选为氮原子上的氢原子)被选自下组的取代基取代：C1-C4烷基、C1-C4卤代烷基、胺基保护基(优选叔丁氧羰基)、卤素。
在另一优选例中，当所述的R3、R4与相邻的氮原子共同构成取代或未取代的5-6元饱和环时，环上的氮原子上可以任选地具有胺基保护基。
在另一优选例中，所述的胺基保护基选自下组：
在另一优选例中，所述的5-7元杂环不是杂芳环。
在另一优选例中，所述的5-7元杂环为饱和杂环，优选5-6元饱和杂环。
在另一优选例中，所述的5-7元杂环仅含有一个或二个杂原子。
在另一优选例中，所述的5-7元杂环中所有的杂原子为N。
在另一优选例中，R3、R4各自独立地选自下组：C1-C4烷基；或R3、R4与相 邻的氮原子共同构成取代或未取代的5-6元饱和环，其中，所述的5-6元饱和环上具有1或2个氮原子，和任选的1个选自下组的杂原子：O。
在另一优选例中，R3、R4不同时为H。
在另一优选例中，R2为环状亚胺。
在另一优选例中，所述的R2为取代或未取代的选自下组的基团：
Figure PCTCN2014090947-appb-000002
Figure PCTCN2014090947-appb-000003
其中，
Figure PCTCN2014090947-appb-000004
表示连接位点；
所述的取代指基团上的一个或多个氢原子被选自下组的取代基取代：C1-C4烷基、卤素。
在另一优选例中，R1为F。
本发明的第二方面，提供了一种N-苄基色胺酮类衍生物，所述衍生物的结构通式如下：
Figure PCTCN2014090947-appb-000005
R1＝H,F
Figure PCTCN2014090947-appb-000006
其中：所述取代基R2为环状亚胺或二烷基取代胺，R1为氢或氟。
在另一优选例中，所述衍生物为选自下组的化合物：
Figure PCTCN2014090947-appb-000007
本发明的第三方面，提供了一种如本发明第一方面或第二方面所述的N-苄基色胺酮类衍生物的制备方法，所述方法包括步骤：
Figure PCTCN2014090947-appb-000008
(d)在惰性溶剂中，在三乙胺存在下，用式5化合物(溴代2-甲基色胺酮)与R2H反应，得到式1化合物，
式中，R1、R2的定义如上。
在另一优选例中，在所述步骤(d)中，所述的惰性溶剂为DMF，较佳地为无水DMF。
在另一优选例中，在所述步骤(d)中，所述反应在约10～40℃下进行。
在另一优选例中，在所述步骤(d)中，所述反应时间为0.5-12小时，较佳地1-5小时。
在另一优选例中，在所述步骤(d)中，所述的式5化合物、R2H和三乙胺的摩尔比为1:(1-2):(2-5)。
在另一优选例中，所述方法在步骤(d)之前还包括步骤(c)：
Figure PCTCN2014090947-appb-000009
(c)在惰性溶剂中，用式4化合物与溴代试剂反应，得到式5化合物；较佳地，所述的溴代试剂为NBS。
在另一优选例中，所述步骤(c)的反应在引发剂存在下进行，较佳地在AIBN存在下进行。
在另一优选例中，所述步骤(c)包括：用NBS与AIBN混合，然后与式4化合物进行反应。
在另一优选例中，步骤(c)中所述的惰性溶剂为二氯甲烷，较佳地为无水二氯甲烷。
在另一优选例中，在所述步骤(c)中，所述反应在75-85℃下进行。
在另一优选例中，在所述步骤(c)中，所述反应时间为15-17小时。
在另一优选例中，在所述步骤(c)中，所述反应中，所述的式4化合物、NBS和AIBN的摩尔比为1:1:(0.005-0.02)。
在另一优选例中，所述方法在步骤(c)之前还包括步骤(b)：
Figure PCTCN2014090947-appb-000010
(b)在惰性溶剂中，在三乙胺存在下，用式3化合物与
Figure PCTCN2014090947-appb-000011
反应，得 到式4化合物。
在另一优选例中，在所述步骤(b)中，所述的惰性溶剂为甲苯，较佳地为无水甲苯。
在另一优选例中，在所述步骤(b)中，所述反应在100～120℃下进行。
在另一优选例中，在所述步骤(b)中，所述反应时间为3.5-4.5小时。
在另一优选例中，在所述步骤(b)中，所述的式3化合物、
Figure PCTCN2014090947-appb-000012
和三乙胺的摩尔比为(0.2-0.5):(0.2-0.5):1。
在另一优选例中，所述方法在步骤(b)之前还包括步骤(a)：
Figure PCTCN2014090947-appb-000013
(a)在惰性溶剂中，用式2化合物与氧化试剂反应，得到式3化合物；较佳地，所述的氧化试剂是间-氯过氧苯甲酸。
在另一优选例中，在所述步骤(a)中，所述的惰性溶剂为二氯甲烷，较佳地为无水二氯甲烷
在另一优选例中，在所述步骤(a)中，所述反应在室温(优选为10～40℃)下进行。
在另一优选例中，在所述步骤(a)中，所述反应时间为1.5-2.5小时。
在另一优选例中，在所述步骤(a)中，所述的式2化合物和氧化试剂的摩尔比为(0.5-1):1。
本发明的第四方面，提供了一种如本发明第二方面所述的N-苄基色胺酮类衍生物的制备方法，所述的合成路线如下：
Figure PCTCN2014090947-appb-000014
R1＝H,F
Figure PCTCN2014090947-appb-000015
具体步骤如下：
(1)甲基靛红酸酐(3)的合成
5-甲基靛红悬浮于干燥二氯甲烷中，在0℃下分批加入间-氯过氧苯甲酸，在室温下搅拌1.5-2.5小时；TLC检测反应完全后，过滤反应形成的白色固体，用乙酸乙酯洗涤三次，得到5-甲基靛红酸酐；其中，5-甲基靛红和间-氯过氧苯甲酸摩尔比为:(0.5-1):1；
(2)2-甲基色胺酮(4)的合成
将甲基靛红酸酐、5-氟靛红和三乙胺的混合物悬浮于干燥甲苯中，在100-120℃下加热3.5-4.5小时，溶剂在减压条件下蒸去，黄色的固体溶解于二氯甲烷中，再加入乙酸乙酯，形成的黄色固体经过滤用乙酸乙酯洗涤三次，得到2-甲基色胺酮；其中,甲基靛红酸酐和5-氟靛红和三乙胺摩尔比为:(0.2-0.5):(0.2-0.5):1；
(3)溴代2-甲基色胺酮(5)的合成
在75-85℃和氮气保护的条件下，将步骤(2)所得的2-甲基色胺酮溶解于干燥的二氯甲烷中，然后分批加入NBS和AIBN的混合物。反应液在75℃加热15-17小时，TLC检测反应完全。反应液冷却到室温后经盐水洗涤，无水硫酸钠干燥，浓缩得到的黄色产物；其中,2-甲基色胺酮和NBS和AIBN摩尔比为:1:1:(0.005-0.02)；
(4)N-苄基色胺酮(1)的合成
将溴代2-甲基色胺酮、脂肪胺和三乙胺在干燥DMF中于室温下搅拌1.5-2.5小时，TLC检测反应完全后，加入50ml水，再用10ml二氯甲烷萃取三次，合并的二氯甲烷相用水洗三次再用无水硫酸钠干燥。浓缩得到的黄色固体用硅胶分离，得到黄色的N-苄基色胺酮衍生物；其中，溴代2-甲基色胺酮和脂肪胺和三乙胺摩尔比为:1:(1-2):(2-5)。
本发明的第五方面，提供了一种如本发明第一或第二方面所述的N-苄基色胺酮衍生物在制备预防和/或治疗具有IDO介导的色氨酸代谢紊乱的病理学特征的疾病的药物中的应用。
本发明的第六方面，提供了一种如本发明第一或第二方面所述的式1化合物或其药学上可接受的盐的用途，用于：
(i)制备IDO抑制剂；
(ii)制备色氨酸代谢紊乱相关疾病的药物；
(iii)体外非治疗性地抑制IDO。
在另一优选例中，所述色氨酸代谢紊乱相关疾病选自下组：肿瘤、抑郁症、焦虑症、艾滋病、自身免疫疾病、心理障碍、莱姆病感染、链球菌感染、神经退行性病症(例如阿尔茨海默病、亨廷顿病和帕金森病)、抑郁症、癌症(包括T细胞白血病和结肠癌)、眼部疾病(例如白内障和与年龄相关的黄化)，或自身免疫性疾病。
本发明的第七方面，提供了一种药物组合物，所述药物组合物包含(i)如本发明第一或第二方面所述的式1化合物，或其药学上可接受的盐；以及(ii)药学上可接受的载体。
在另一优选例中，所述的药物组合物用于治疗与色氨酸代谢紊乱相关的疾病。
在另一优选例中，所述色氨酸代谢紊乱相关疾病选自下组：肿瘤、抑郁症、 焦虑症、艾滋病、自身免疫疾病、心理障碍、莱姆病感染、链球菌感染、神经退行性病症(例如阿尔茨海默病、亨廷顿病和帕金森病)、抑郁症、癌症(包括T细胞白血病和结肠癌)、眼部疾病(例如白内障和与年龄相关的黄化)，或自身免疫性疾病。
本发明的第八方面，提供了一种IDO抑制剂，所述的IDO抑制剂包含如本发明第一或第二方面所述的式1化合物，或其药学上可接受的盐。
本发明的第九方面，提供了一种体外非治疗性地抑制IDO活性的方法，所述方法包括步骤：使抑制有效量的如本发明第一或第二方面所述的式1化合物，或其药学上可接受的盐与待抑制对象接触。
本发明的第十方面，提供了一种制备治疗色氨酸代谢紊乱相关疾病的药物组合物的方法，所述方法包括：将治疗有效量的如本发明第一或第二方面所述的式1化合物或其药学上可接受的盐与药学上可接受的载体混合，从而形成药物组合物。
本发明的第十一方面，提供了一种治疗色氨酸代谢紊乱相关疾病的方法，所述方法包括：对治疗对象施用治疗有效量的如本发明第一或第二方面所述的式1化合物，或其药学上可接受的盐。
在另一优选例中，所述色氨酸代谢紊乱相关疾病选自下组：肿瘤、抑郁症、焦虑症、艾滋病、自身免疫疾病、心理障碍、莱姆病感染、链球菌感染、神经退行性病症(例如阿尔茨海默病、亨廷顿病和帕金森病)、抑郁症、癌症(包括T细胞白血病和结肠癌)、眼部疾病(例如白内障和与年龄相关的黄化)，或自身免疫性疾病。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
本发明人经过长期而深入的研究，意外地发现，具有如式1所示结构的一类N-苄基色胺酮类衍生物具有相当优异的IDO抑制活性，所述化合物与一些和其结构非常相近的N-苄基色胺酮类衍生物相比，IDO抑制活性(IC50)居然有极其显著的改善(IC50值可降低1-3个数量级)，因此具有很好的应用前景。基于上述发现，发明人完成了本发明。
术语
除特别说明之处，本发明的所有化合物均包括所有光学异构体或互变异构体形式。
术语“C1～C4烷基”指具有1～4个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、或类似基团。
术语“C2～C4烯基”指具有2～4个碳原子的直链或支链烯基，例如乙烯基、丙烯基、丁烯基、或类似基团。
术语“C2～C4炔基”指具有2～4个碳原子的直链或支链炔基，例如乙炔基、丙炔基、丁炔基、或类似基团。
术语“5-7元杂环”指具有指5至7元的单环，所述的环不具有完全共轭的π电子系统。特别地，在本发明中，所述的环上可以任意地具有1-3个杂原子，所述的杂原子包括O或N。代表性的饱和环例子包括：哌嗪基、吗啡啉基等。
除非特别说明，术语“取代”指基团上的一个或多个氢原子被选自下组的取代基进行取代：C1-C4烷基、C1-C4卤代烷基、胺基保护基(如叔丁氧羰基)、卤素。
术语“卤素”指F、Cl、Br和I。
N-苄基色胺酮类衍生物
色胺酮为吲哚喹唑啉类生物碱，其化学名称为吲哚[2,1-b]喹唑啉-6,12-二酮。色胺酮是一种黄色针状结晶，主要存在于马蓝、蓼蓝、菘蓝等产蓝植物中。另外，也可以从微生物的发酵液中提取。
虽然从产蓝植物及微生物的代谢产物中可提取色胺酮，但其分离过程长、提取率低，难以满足研究和临床用药的需求。只有通过探索耗时短、收率高、简便易得的人工合成途径才能为色胺酮的应用提供更多的资源，使其进一步的开发和应用成为可能。
为了克服现有技术的不足，本发明对色胺酮进行了结构改造，以改善色胺酮的溶解性能和药理活性，目的在于获得有应用价值的活性化合物。本发明的研究和药理试验表明，在色胺酮分子中引入水溶性基团而形成的N-苄基色胺酮衍生物可以作为更高效的IDO抑制剂，其具有抗菌、抗炎、抗肿瘤等多种药理活性，有广阔的应用前景。此外，相比已有的色胺酮衍生物合成方法而言，本发明的合成方法具有操作简单、条件温和、收率高等优点，更易于工业化生产。
具体地，本发明的N-苄基色胺酮类衍生物具有如下式1所示的结构通式：
Figure PCTCN2014090947-appb-000016
式中
R1为氢或氟；
R2为-NR3R4；
所述的R3、R4各自独立地选自下组：C1-C4烷基；或R3、R4与相邻的氮原子共同构成取代或未取代的5-6元饱和环，其中，所述的5-6元饱和环上具有至少一个氮原子，和任选的1-2个选自下组的杂原子：O、N；
所述的取代指基团上的一个或多个氢原子(优选为氮原子上的氢原子)被选自下组的取代基取代：C1-C4烷基、叔丁氧羰基、卤素。
优选地，所述的R2上具有1个氮原子，以及1个选自下组的杂原子：O、N。
优选地，所述的R2为取代或未取代的选自下组的基团：
Figure PCTCN2014090947-appb-000017
Figure PCTCN2014090947-appb-000018
其中，
Figure PCTCN2014090947-appb-000019
表示连接位点；
所述的取代指基团上的一个或多个氢原子被选自下组的取代基取代：C1-C4烷基、卤素。
本发明所述的化合物、其互变异构形式、其结构类似物或其药物可接受的盐，以及含有至少一种该化合物、其结构类似物或其药物可接受的盐的组合物均可以用于抑制IDO，以及在治疗和/或预防具有IDO介导的色氨酸代谢途径的病理学特征的疾病中的用途。这样的疾病包括但不限于，肿瘤、癌症、眼部疾病、自身免疫性疾病、心理障碍、抑郁症和焦虑症。所述用途包括体内和体外应用，以及在制备药物、IDO抑制剂和药物组合物中的用途。特别地，本发明人发现，在一种特别优选的实施例中，当R1为F时，所述的化合物具有最佳的IDO抑制活性。
N-苄基色胺酮类衍生物的制备
近年来，药物化学家一直致力于色胺酮及其衍生物的合成研究，合成色胺酮的主要方法是吲哚醌与靛红酸酐反应，该方法简单收率高、反应条件温和。另外，可以在吲哚醌与靛红酸酐两种原料母体上引入功能化基团，可以合成各种功能化色胺酮。目前，合成吲哚醌最主要的方法是利用水合三氯乙醛、羟胺和苯胺在盐酸水溶液中反应生成肟类化合物，然后在浓硫酸作用下闭环得到吲哚醌，该方法非常适用于合成含卤素和烷基的色胺酮，收率也较高。但该方法难以合成含活性基团的色胺酮，这是由于在合成吲哚醌的过程中，这些基团易于发生多种副反应。
本发明人经过长期的研究，成功得到了一种制备具有活性基团的色胺酮的合成方法。具体地，所述的方法包括步骤：
Figure PCTCN2014090947-appb-000020
(d)在惰性溶剂中，在三乙胺存在下，用式5化合物(溴代2-甲基色胺酮)与R2H反应，得到式1化合物。
在另一优选例中，所述方法还包括步骤：
Figure PCTCN2014090947-appb-000021
(c)在惰性溶剂中，用式4化合物(2-甲基色胺酮)与溴代试剂反应，得到式5化合物；较佳地，所述的溴代试剂为NBS。
在本发明的一个优选实施例中，所述的溴代反应在引发剂，例如AIBN存在下进行。优选的实施方式包括：用NBS与AIBN混合，然后与式4化合物进行反 应。
在另一优选例中，所述方法还包括步骤：
Figure PCTCN2014090947-appb-000022
(b)在惰性溶剂中，在三乙胺存在下，用式3化合物与
Figure PCTCN2014090947-appb-000023
反应，得到式4化合物。
在另一优选例中，所述方法还包括步骤：
Figure PCTCN2014090947-appb-000024
(a)在惰性溶剂中，用式2化合物与氧化试剂反应，得到式3化合物；较佳地，所述的氧化试剂是间-氯过氧苯甲酸。
上述各步骤的反应时间，反应温度等可以根据实际情况加以调整，例如采用本领域常用的方法(如TLC法)确定反应终点。各步骤的溶剂没有特别的限制，可以选用本领域常用的，不与反应物发生反应的惰性溶剂。应理解，在本申请公开了上述反应路线后，各步骤的具体条件是本领域技术人员结合现有技术中的知识可以确定的。
本发明的N-苄基色胺酮类衍生物的制备方法中，一种最佳的合成路线如下：
Figure PCTCN2014090947-appb-000025
(1)甲基靛红酸酐(3)的合成
5-甲基靛红悬浮于干燥二氯甲烷中，在0℃下分批加入间-氯过氧苯甲酸，在室温下搅拌1.5-2.5小时；TLC检测反应完全后，过滤反应形成的白色固体，用乙酸乙酯洗涤三次，得到5-甲基靛红酸酐；其中,5-甲基靛红和间-氯过氧苯甲酸摩尔比为:0.5-1:1。
(2)2-甲基色胺酮(4)的合成
将甲基靛红酸酐、5-氟靛红和三乙胺的混合物悬浮于干燥甲苯中，在100-120℃下加热3.5-4.5小时，溶剂在减压条件下蒸去，黄色的固体溶解于二氯甲烷中，再加入乙酸乙酯，形成的黄色固体经过滤用乙酸乙酯洗涤三次，得到2-甲基色胺酮；其中,甲基靛红酸酐和5-氟靛红和三乙胺摩尔比为:0.2-0.5:0.2-0.5:1。
(3)溴代2-甲基色胺酮(5)的合成
在75-85℃和氮气保护的条件下，将步骤(2)所得的2-甲基色胺酮溶解于干燥的二氯甲烷中，然后分批加入NBS和AIBN的混合物。反应液在75℃加热15-17小时，TLC检测反应完全。反应液冷却到室温后经盐水洗涤，无水硫酸钠干燥，浓缩得到的黄色产物；其中,2-甲基色胺酮和NBS和AIBN摩尔比为:1:1:0.005-0.02。
(4)N-苄基色胺酮(1)的合成
将溴代2-甲基色胺酮、脂肪胺和三乙胺在干燥DMF中于室温下搅拌1.5-2.5小时，TLC检测反应完全后，加入50ml水，再用10ml二氯甲烷萃取三次，合并的二氯甲烷相用水洗三次再用无水硫酸钠干燥。浓缩得到的黄色固体用硅胶分离，得到黄色的N-苄基色胺酮衍生物。其中,溴代2-甲基色胺酮和脂肪胺和三乙胺摩尔比为:1:1-2:2-5。
上述各式中，各基团的定义如上文中所述。
IDO抑制剂及其用途
由于本发明的N-苄基色胺酮衍生物具有十分优异的IDO抑制活性，因此可以用于制备预防和/或治疗具有IDO介导的色氨酸代谢紊乱的病理学特征的疾病的药物(如肿瘤等)。
所述的色氨酸代谢紊乱疾病可以是任何本领域已知或未知的具有IDO介导的色氨酸代谢紊乱的病理学特征的疾病，优选地，所述色氨酸代谢紊乱相关疾病选自下组：肿瘤、抑郁症、焦虑症、艾滋病、自身免疫疾病、心理障碍、莱姆病感染、链球菌感染、神经退行性病症(例如阿尔茨海默病、亨廷顿病和帕金森病)、抑郁症、癌症(包括T细胞白血病和结肠癌)、眼部疾病(例如白内障和与年龄相关的黄化)，或自身免疫性疾病。
与现有技术相比，本发明的主要优点包括：
(1)本发明提供了一类结构新颖的IDO抑制剂，与结构接近的化合物相比，该类抑制剂具有令人意外的活性改善。所述化合物既具有IDO抑制活性，同时也可以作为进一步修饰的药物中间体，具有开发成新的治疗癌症药物的潜在应用价值。
(2)本发明还提供了一类合成的N-苄基色胺酮衍生物的方法，该合成路线具有操作简便、反应条件温和、节省溶剂、减少污染等优点，便于工业化生产。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通 常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。如无特别说明，以下实施例中所涉及到的试剂或原料均为市售。具体合成路线如下：
Figure PCTCN2014090947-appb-000026
各步骤反应条件：
步骤a.m-CPBA(2eq.),CH2Cl2,RT；
步骤b.Et3N,甲苯,回流,75％；
步骤c.NBS,AIBN,CH2Cl2,回流；
步骤d.K2CO3,KI,DMF,胺。
实施例12-((二甲基氨基)甲基)-8-氟吲哚并[2,1-b]喹唑啉-6,12-二酮(1a)的合成
具体步骤如下：
步骤(1)、6-甲基-1H-苯并[d][1,3]恶嗪-1,4-二酮(3a)的合成
Figure PCTCN2014090947-appb-000027
化合物2a(500mg,3mmol)悬浮于10ml的干燥二氯甲烷中，在0℃下分批加入间-氯过氧苯甲酸(1.3g,6mmol,75％)。在室温条件下搅拌2小时后。TLC检测反应完全后，过滤反应形成的白色固体，然后用10ml乙酸乙酯洗涤三次后，得到化合物3a(350mg,65％)。
步骤(2)、8-氟-2-甲基吲哚并[2,1-b]喹唑啉-6,12-二酮(4a)的合成
Figure PCTCN2014090947-appb-000028
化合物3a(1g,5.6mmol),5-氟靛红(0.93g,5.6mmol)和三乙胺(1.5ml,11.2mmol)的混合物悬浮于干燥的甲苯(10ml)中，在110℃下加热4小时。减压蒸去溶剂，所获黄色固体溶于2ml的二氯甲烷中，然后再加入2ml乙酸乙酯，形成黄色固体经过滤，用2ml乙酸乙酯洗涤三次，得到黄色固体化合物4a(1.1g, 75％)。
表征数据：
1H NMR(400MHz,DMSO)δ8.50(dd,J＝8.8,4.2Hz,1H),8.14(s,1H),7.86(d,J＝8.2Hz,1H),7.82–7.76(m,1H),7.76–7.67(m,1H),7.44–7.37(m,1H),2.53(s,3H).
步骤(3)、2-(溴甲基)8-氟吲哚[2,1-b]喹唑啉-6,12-二酮(5a)的合成
Figure PCTCN2014090947-appb-000029
在80℃和氮气保护的条件下，向化合物4a(500mg,1.78mmol)的二氯甲烷(3.6ml)溶液中分三批加入NBS(381mg,2.14mmol)和AIBN(29mg,0.18mmol)的混合物。反应液在80℃加热16小时。TLC检测反应完全，反应液冷至室温，盐水洗涤，无水硫酸钠干燥，浓缩得到黄色产物5a。
表征数据：
1H NMR(400MHz,CDCl3)δ8.66(dd,J＝8.8,4.0Hz,1H),8.45(d,J＝1.9Hz,1H),8.04(d,J＝8.3Hz,1H),7.90(dd,J＝8.3,2.1Hz,1H),7.61(dd,J＝6.5,2.6Hz,1H),7.52(td,J＝8.6,2.7Hz,1H),4.65(s,2H)。
步骤(4)、2-((二甲基氨基)甲基)-8-氟吲哚并[2,1-b]喹唑啉-6,12-二酮(1a)的合成
Figure PCTCN2014090947-appb-000030
化合物5a(500mg,1.39mmol),盐酸二甲胺(Dimethylamine hydrochloride)(227mg,2.78mmol),碘化钾(10mg)和三乙胺(0.5ml)在5ml的DMF中在室温搅拌2小时。TLC检测反应完全后，加入50ml水，用10ml乙酸乙酯萃取三次后，合并的有机相用水洗三次，再用无水硫酸钠干燥。浓缩得到的黄色固体用硅胶柱分离后得到黄色的化合物1a。
表征数据：
1HNMR(400MHz,CDCl3)δ8.63(dd,J＝8.8,4.1Hz,1H),8.33(d,J＝1.5Hz,1H),7.98(d,J＝8.3Hz,1H),7.87(dd,J＝8.3,1.8Hz,1H),7.58(dd,J＝6.5,2.6Hz,1H),7.48(td,J＝8.7,2.7Hz,1H),3.61(s,2H),2.30(s,6H)。
实施例24-((8-氟-6,12-二氧代-6,12-二羟基吲哚并[2,1-b]喹啉-2-基)甲基)哌嗪-1-甲酸叔丁酯(1b)的合成
步骤(1)-步骤(3)同实施例1。
步骤(4)：4-((8-氟-6,12-二氧代-6,12-二羟基吲哚并[2,1-b]喹啉-2-基)甲基)哌嗪-1-甲酸叔丁酯(1b)的合成
Figure PCTCN2014090947-appb-000031
化合物5a(500mg,1.39mmol),N-Boc哌嗪(N-Bocpiperazine)(546mg,2.78mmol),碘化钾(10mg)和三乙胺(0.5ml)在5ml的DMF中，于室温下搅拌2小时。TLC检测反应完全后，加入50ml水，用10ml乙酸乙酯萃取三次，合并有机相用水洗三次，再用无水硫酸钠干燥。浓缩得到的黄色固体用硅胶柱分离后得到黄色化合物1b。
表征数据：
1H NMR(400MHz,CDCl3)δ8.66(dd,J＝8.8,4.0Hz,1H),8.39(d,J＝1.5Hz,1H),8.01(d,J＝8.3Hz,1H),7.90(dd,J＝8.3,1.8Hz,1H),7.60(dd,J＝6.5,2.6Hz,1H),7.51(td,J＝8.6,2.7Hz,1H),3.71(s,2H),3.47(m,4H),2.46(m,4H),1.48(s,9H)。
实施例38-氟-2-(哌嗪-1-基甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮(1c)的合成
Figure PCTCN2014090947-appb-000032
步骤(1)-步骤(3)同实施例1。
步骤(4)：8-氟-2-(哌嗪-1-基甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮(1c)的合成
化合物5a(500mg,1.39mmol),N-叔丁氧羰基哌嗪(N-Bocpiperazine)(546mg,2.78mmol),碘化钾(10mg)和三乙胺(0.5ml)在5ml的DMF中于室温搅拌2小时。TLC检测反应完全后，加入50ml水，用10ml乙酸乙酯萃取三次，合并有机相用水洗三次，再用无水硫酸钠干燥。浓缩得到的黄色固体。将黄色固体溶解于3毫升的二氯甲烷中，在室温条件下加入三氟乙酸1毫升。反应液在室温搅拌1小时后，TLC(CH2Cl2/MeOH＝10/1,Rf 0.2)检测反应完全后，溶剂在减压条件下去除，加入保护NaHCO3溶液10毫升，用乙酸乙酯(10ml×3)萃取后，有机相用盐水洗涤，干燥，然后浓缩得到固体用硅胶柱分离得到黄色固体化合物1c。
表征数据：
1H NMR(400MHz,CDCl3)δ8.65(dd,J＝8.8,4.0Hz,1H),8.37(s,1H),8.00(d,J＝8.3Hz,1H),7.89(dd,J＝8.3,1.6Hz,1H),7.59(dd,J＝6.5,2.6Hz,1H),7.50(td,J＝8.7,2.7Hz,1H),3.70(s,2H),3.51–3.44(m,4H),2.46(s,4H)。
实施例4：8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮(1d)的合成
Figure PCTCN2014090947-appb-000033
步骤(1)-步骤(3)同实施例1。
步骤(4)：8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮(1d)的合成
化合物5a(220mg,0.61mmol),N-甲基哌嗪(N-Methylpiperazine)(122mg,1.22mmol),碘化钾(10mg)和三乙胺(0.5ml)在5ml的DMF中在室温搅拌2小时。TLC检测反应完全后，加入50ml水，用10ml乙酸乙酯萃取三次，合并有机相用水洗三次，再用无水硫酸钠干燥。浓缩得到的黄色固体用硅胶柱分离后得到黄色化合物1d。
表征数据：
1H NMR(400MHz,CDCl3)δ8.65(dd,J＝8.8,4.0Hz,1H),8.38(d,J＝1.5Hz,1H),7.99(d,J＝8.3Hz,1H),7.87(dd,J＝8.3,1.8Hz,1H),7.59(dd,J＝6.5,2.6Hz,1H),7.50(td,J＝8.7,2.7Hz,1H),3.50(s,2H),2.59(s,8H),2.38(s,3H)。
实施例58-氟-2-((吗啡啉-1-基)甲基)吲哚并[2,1-b)喹唑啉-6，12-二酮(1e)的合成
步骤(1)-步骤(3)同实施例1。
步骤(4)：8-氟-2-((吗啡啉-1-基)甲基)吲哚并[2,1-b)喹唑啉-6，12-二酮(1e)的合成
Figure PCTCN2014090947-appb-000034
化合物5a(500mg,1.39mmol),吗啡啉(242mg,2.78mmol),碘化钾(10mg)和三乙胺(0.5ml)在5ml DMF中在室温搅拌2小时。TLC(EtOAc,Rf 0.5)检测反应完全后，加入50ml水，用10ml乙酸乙酯萃取三次，合并有机相水洗三次，用无水硫酸钠干燥，浓缩得到黄色固体用硅胶柱(EtOAc)分离后得到黄色化合物1e。
表征数据：
1H NMR(400MHz,CDCl3)δ8.66(dd,J＝8.8,4.1Hz,1H),8.39(s,1H),8.01(d,J＝8.3Hz,1H),7.90(dd,J＝8.3,1.7Hz,1H),7.60(dd,J＝6.5,2.7Hz,1H),7.51(td,J＝8.6,2.7Hz,1H),3.79–3.73(m,4H),3.70(s,2H),2.52(s,4H)。
实施例62-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮(1f)的合成
步骤(1)同实施例1，
步骤(2)、2-甲基吲哚[2,1-b]喹唑啉-6,12-二酮(4b)的合成
Figure PCTCN2014090947-appb-000035
化合物3a(1.7g,9.6mmol),靛红(1.4g,9.6mmol)和三乙胺(2.7ml,19.2mmol)的混合物悬浮于干燥的甲苯(18ml)中，在110℃下加热4小时。反应液冷至室温后，形成的黄色固体经过滤，用2ml乙酸乙酯洗涤三次，得到黄色固体化合物4b(0.5g,20％)。
表征数据：
1H NMR(400MHz,CDCl3)δ8.65(d,J＝8.1Hz,1H),8.25(s,1H),7.94(dd,J＝7.8,5.0Hz,2H),7.85–7.75(m,1H),7.71–7.64(m,1H),7.44(t,J＝7.5Hz,1H),2.58(s,3H)。
步骤(3)、2-甲基吲哚[2,1-b]喹唑啉-6,12-二酮(5b)的合成
Figure PCTCN2014090947-appb-000036
在80℃和氮气保护的条件下，向化合物4b(262mg,1.0mmol)的二氯甲烷(5ml)溶液中分三批加入NBS(381mg,2.14mmol)和AIBN(29mg,0.18mmol)的混合物。反应液冷却到室温，盐水洗涤，无水硫酸钠干燥，浓缩后得到的黄色产物5b(245mg,91％)。
步骤(4)、2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮(1f)的合成
Figure PCTCN2014090947-appb-000037
化合物5b(220mg,0.61mmol),N-甲基哌啶(116mg,1.616mmol),碘化钾(10mg)和三乙胺(0.5ml)在5ml DMF中于室温下搅拌2小时。TLC检测反应完全后，加入50ml水，用10ml乙酸乙酯萃取三次，合并有机相用水洗三次，用无水硫酸钠干燥。浓缩得到的黄色固体用硅胶柱分离得到黄色化合物1f(120mg,55％)。
表征数据：
1H NMR(400MHz,CDCl3)δ8.66(d,J＝8.1Hz,1H),8.39(s,1H),8.01(d,J＝8.3Hz,1H),7.94(d,J＝7.5Hz,1H),7.89(dd,J＝8.2,1.7Hz,1H),7.82(t,J＝7.8Hz,1H),7.45(t,J＝7.5Hz,1H),3.71(s,2H),2.71–2.41(m,8H),2.33(s,3H)。
实施例7IDO抑制活性检测
含人IDO基因的质粒的构建、在大肠杆菌中的表达、提取及纯化均按Littlejohn等报道的方法进行(Takikawa O,Kuroiwa T,Yamazaki F,et al.J. Biol.Chem.1988，263,2041-2048)。各化合物对IDO的抑制活性按文献介绍的方法检测。在96孔板上将50mM磷酸钾缓冲液(pH 6.5)，40mM维生素C，400μg/ml过氧化氢酶，20μM亚甲基蓝和IDO酶混合。向上述混合液内加入底物L-色氨酸和待测样品。反应在37℃下进行60分钟,加入30％(w/v)三氯乙酸使反应终止。96孔板在65℃下加热15分钟，使之完成从甲酰犬尿氨酸到犬尿氨酸的转化，然后6000g旋转5分钟。每孔取出100μl上清液转移到新的96孔板内，加入2％(w/v)对-(二甲基氨基)苯甲醛的乙酸溶液，犬尿氨酸与之反应产成的黄颜色可使用酶标仪在490nm观测，实验结果如表1所示。
实施例8是否为可逆抑制剂的判定
在固定抑制剂浓度的情况下，用一系列不同浓度的酶与抑制剂反应并测定反应速度。以反应速度对酶浓度(ν～[E])作图，根据曲线的特征可以判定是否是可逆抑制剂。
反应条件：在500μl的反应体系中，先加入50mM磷酸钾缓冲液(pH 6.5)，40mM维生素C，400μg/ml过氧化氢酶，20μM亚甲基蓝，300mM底物L-色氨酸或同时加入100mM抑制剂，混合液37℃保温5分钟，再向上述混合液内分别加入不同体积的IDO酶，反应在37℃下进行30分钟，加入30％(w/v)三氯乙酸200μl使反应终止，反应体系在65℃加热15分钟，使之完成从甲酰犬尿氨酸到犬尿氨酸的转化，然后12000rpm旋转10分钟，取上清与等体积2％(w/v)对-二甲氨基苯甲醛的乙酸溶液混合，用酶标仪检测490nm波长读数。以ν～[E]作图，实验结果如表1所示。
实施例9抑制剂类型判断及Ki值测定
在500μl的反应体系中，先加入50mM磷酸钾缓冲液(pH 6.5)，40mM维生素C，400μg/ml过氧化氢酶，20μM亚甲基蓝，分别加入100、250、300mM底物L-色氨酸，在一个底物浓度下，向各管反应体系中分别加入不同浓度的化合物，混合液37℃保温5分钟，再向上述混合液内加入10μl IDO(约20nM)，反应在37℃下进行30分钟，加入30％(w/v)三氯乙酸200μl使反应终止，反应体系在65℃水浴加热15分钟，使之完成从甲酰犬尿氨酸到犬尿氨酸的转化，然后12000rpm离心10分钟，取上清与等体积2％(w/v)对-二甲氨基苯甲醛的乙酸溶液混匀反应，用酶标仪检测490nm波长读数。以Dixon作图法(1/v～[I])判定化合物的抑制剂类型，以S/v～[I]作图，得到抑制剂的Ki值，实验结果如表1所示。
实施例10半数有效抑制浓度IC50的测定
先将50mM磷酸钾缓冲液(pH 6.5)，40mM维生素C，400μg/ml过氧化氢酶，20μM亚甲基蓝，底物L-色氨酸150mM和抑制剂混合。抑制剂浓度选用100，200，400，600，800，1000，1200μM混合液37℃保温5分钟，再向上述混合液内加入IDO酶。反应在37℃下进行30分钟，加入30％(w/v)三氯乙酸200μl使反 应终止，反应体系在65℃加热15分钟，使之完成从甲酰犬尿氨酸到犬尿氨酸的转化，然后12000rpm旋转10分钟，取200μl上清与等体积2％(w/v)对-二甲氨基苯甲醛的乙酸溶液混合，犬尿氨酸与之反应产生的黄颜色可使用酶标仪在490nm检测，所得结果利用IC50计算软件计算,得到抑制剂的IC50值，实验结果如表1所示。
Figure PCTCN2014090947-appb-000038
实施例11半数有效抑制浓度IC50(细胞)的测定
利用脂质体Lipofectamin 2000介导质粒pcDNA3.1-hIDO瞬转HEK 293细胞。在细胞水平抑制剂活性测定时，HEK293细胞培养基为高糖DMEM，含50U/mL青霉素，50U/mL链霉素，10％FBS，37℃，5％CO2培养。细胞转染质粒24h后，加入待测药物，孵育一段时间后，取上清到另一96孔板中，加入10μL 30％(w/v)三氯乙酸，在65℃加热15min使之完成甲酰犬尿氨酸向犬尿氨酸的转化，然后12000rpm离心10min，取等体积2‰(w/v)对二甲氨基苯甲醛的乙酸溶液混合显色，最后采用酶标仪在490nm下检测吸光值，实验结果如表1所示。
利用上述实施例7-11的方法，对实施例1-6制备的化合物的IDO抑制活性进行测定，并用目前体内、外实验中通用的IDO抑制剂1-甲基色氨酸(1-MT，市售)作为对照物，测定结果如表1所示。
表1上述实施例中合成的N-苄基色胺酮衍生物(I)的IDO抑制活性
Figure PCTCN2014090947-appb-000039
实施例12T细胞增殖实验
12.1脾脏淋巴细胞分离(按照小鼠淋巴细胞分离液说明操作)：
1.断颈处死2只小鼠，在超净台中取出脾脏，放在装有RP1640培养基的6cm板中待用。
2.在50mL离心管中放入100μm筛网，脾脏用剪刀剪碎，在筛网上研磨，加少量RP1640可起到保护作用。
3.加入研磨液2倍体积的淋巴细胞分离液。
4.800g水平离心20min，离心结束可看到液体分3层，中间层略带黄色。
5.吸取中间层细胞，加入RP1640培养基，颠倒洗涤。室温，250g离心10min收集细胞。
6.吸去培养基，用RP1640培养基重悬细胞，细胞计数。
12.2LLC(Lewis lung carcinoma，Lewis肺癌)细胞处理：
1.吸去培养基(高糖DMEM，10％FBS)，PBS洗1-2次；
2.加入0.25％胰酶消化；
3.吸去胰酶，加入培养基，将细胞吹打下来，转移至1.5mL离心管中；
4.离心，吸去上清，加入1mL DMEM培养基重新悬浮细胞；
5.加入丝裂霉素C(终浓度25μg/mL)，吹打混匀，37℃，水浴30min；
6.RP1640洗3次，细胞计数，待用。
12.3实验步骤
1.将处理过的LLC细胞2×104个/孔(刺激细胞)，脾脏淋巴细胞105个/孔(反应细胞)，加入96孔板，加入RP1640(10％FBS)，补齐至200μL；
2.分组，给药组加入50μM IDO抑制剂，置于37℃、湿度95％、5％CO2的培养箱中培养，培养72h；
3.WST-1试剂盒测定T细胞增殖情况,酶标仪测定450nm波长处吸光度值；
4.计算T淋巴细胞增殖率：
T淋巴细胞增殖率(％)＝[给药孔(T淋巴细胞+LLC细胞+IDO抑制剂)OD值
-对照孔(T淋巴细胞+LLC细胞)OD值]/对照孔(T淋巴细胞+LLC细胞)OD值×100％
12.4实验结果
表2上述实施例中合成的N-苄基色胺酮衍生物(I)刺激T细胞增殖的作用
  1-MT 实施例1 实施例2 实施例3 实施例4 实施例5 实施例6
T细胞增殖率 4.88％ 38.48 40.64 41.24 50.27 39.40 13.84
实施例138-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮(实施例4)抗肿瘤作用
1.LLC Lewis肿瘤模型建立：健康雌性C57BL/6小鼠，体重20±1g，购自上海斯莱克动物实验中心，SPF级实验室喂养。用无菌生理盐水将Lewis肺癌细胞浓度为1×107/mL细胞悬液。在无菌操作下在小鼠腋下皮下注射，每只接种0.2mL。
2.实验分组及预防给药：小鼠随机分为5组，每组10只。对照组给以等量 0.5％羧甲基纤维素钠，灌胃给药；1-甲基色氨酸(1-MT)高剂量组为150mg/kg，低剂量组为75mg/kg，灌胃给药；8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮高剂量组为150mg/kg，低剂量组为75mg/kg，灌胃给药。连续给药7天后接种肿瘤，之后继续给药21天并观察肿瘤生长情况。从接瘤当天开始测量肿瘤长径(a)和短径(b)，每隔一天测量一次，肿瘤体积＝ab^2/2。停药后次日断颈处死小鼠，剥瘤称重，液氮保存。
3.预防给药治疗结果：小鼠经预防给药一周及接种后继续给药治疗，给与8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮组的小鼠体内Lewis肺癌肿瘤的生长明显受到抑制，高剂量给药开始后第20天依然没有明显的肿瘤生成，而对照组于给药后第11天发现肿瘤，1-MT组于给药后第13天发生肿瘤。从肿瘤体积角度看，8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮和1-MT高剂量给药的肿瘤抑制率分别为62.15％和32.35％，低剂量给药的肿瘤抑制率分别为46.62％和27.00％。从肿瘤重量角度，高剂量8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮和1-MT的肿瘤抑制率分别为57.8％和28.2％。8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮给药组小鼠体重正常、皮毛有光泽、反应较灵活。
4.实验分组及接瘤后给药：小鼠于接种肿瘤48h后，随机分为4组，每组10只。当肿瘤直径达到5mm时，首次给药。对照组给以等量0.5％CMC(羧甲基纤维素钠)，灌胃给药；1-MT组，给药量为200mg/kg，灌胃给药；8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮组，给药量为200mg/kg，灌胃给药；紫杉醇组，给药量为100mg/kg，尾静脉注射，每周一次；8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮+紫杉醇组，8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮给药量200mg/kg，，紫杉醇组给药量为100mg/kg(尾静脉注射)，连续给药两周。从给药当天开始测量肿瘤长径(a)和短径(b)，每隔一天测量一次，肿瘤体积＝ab^2/2。停药后次日断颈处死小鼠，剥瘤称重，液氮保存。
5.接瘤后给药治疗结果：小鼠经200mg/kg的8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮或1-MT连续灌胃给药两周后，体内Lewis肺癌肿瘤的生长与对照组相比明显受到抑制，并且8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮组抑制肿瘤生长的效果比1-MT组更好。从肿瘤体积角度，8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮、1-MT、紫杉醇、8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮+紫杉醇的抑制率分别为23.38％、15.68％、51.45％、72.48％。紫杉醇作为一种临床上常用的化疗药，虽然几乎能抑制肿瘤的生长，但存在较强的毒副作用，实验过程中发现小鼠体形消瘦、皮毛疏松、少动现象。而8-氟-2-((4-甲基哌嗪-1-基)甲基)吲哚并[2,1-b]喹唑啉-6,12-二酮单独给药尽管不能引起肿瘤消退，但能较大程度抑制肿瘤生长，并且该组小鼠体重正常、皮毛有光泽、反应较灵活。
对比例2-甲基吲哚[2,1-b]喹唑啉-6,12-二酮的制备及活性测试
步骤(1)同实施例1。
步骤(2)、2-甲基吲哚[2,1-b]喹唑啉-6,12-二酮(4b)的合成
Figure PCTCN2014090947-appb-000040
化合物3a(1.7g,9.6mmol),靛红(1.4g,9.6mmol)和三乙胺(2.7ml,19.2mmol)的混合物悬浮于干燥的甲苯(18ml)中，在110℃下加热4小时。反应液冷至室温后，形成的黄色固体经过滤，用2ml乙酸乙酯洗涤三次，得到黄色固体化合物4b(0.5g,20％)。
表征数据：
1H NMR(400MHz,CDCl3)δ8.65(d,J＝8.1Hz,1H),8.25(s,1H),7.94(dd,J＝7.8,5.0Hz,2H),7.85–7.75(m,1H),7.71–7.64(m,1H),7.44(t,J＝7.5Hz,1H),2.58(s,3H)。
用如实施例9-11所述的方法测定对比例所得化合物的活性，IDO抑制活性体外IC50为997.25uM，Ki及体内IC50未测出。
从表1和上述实验数据中可以看出，与现有技术中的IDO抑制剂，以及具有类似结构的化合物相比，本申请的化合物具有异常优异的IDO抑制活性。具体地，相较于现有技术中已知的IDO抑制剂1-MT，本申请化合物的体外和细胞内IC50值都降低了1-3个数量级，这说明本申请化合物具有十分优异的IDO抑制活性。
值得注意的是，本申请的化合物和一些与其结构十分相近的化合物(如对比例化合物)相比，抑制活性方面的改进非常显著。甚至与一些结构中同样具有活性基团的IDO抑制剂相比，其抑制活性也具有相当大的提高。相较于本领域常规的化合物结构与活性的相关性，本申请化合物与其类似结构在生物活性方面的区别是一种罕见的例外情况。这说明，本申请化合物是一类相当独特的结构，其生物活性是令人意外的。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (15)

  1. 一种N-苄基色胺酮类衍生物，其特征在于，所述衍生物具有如下式1所示的结构通式：
    Figure PCTCN2014090947-appb-100001
    式中
    R1为氢或氟；
    R2为-NR3R4；
    所述的R3、R4各自独立地选自下组：H、取代的或未取代的C1-C4烷基、取代的或未取代的C2-C4烯基、取代的或未取代的C2-C4炔基、取代的或未取代的C3-C6环烷基；
    或R3、R4与相邻的氮原子共同构成取代或未取代的5-7元杂环，其中，所述的5-6元饱和环上具有1-2个氮原子，和0-2个选自下组的杂原子：O、S；
    所述的取代指基团上的一个或多个氢原子(优选为氮原子上的氢原子)被选自下组的取代基取代：C1-C4烷基、C1-C4卤代烷基、胺基保护基(优选叔丁氧羰基)、卤素。
  2. 如权利要求1所述的N-苄基色胺酮类衍生物，其特征在于，R3、R4各自独立地选自下组：C1-C4烷基；或R3、R4与相邻的氮原子共同构成取代或未取代的5-6元饱和环，其中，所述的5-6元饱和环上具有1或2个氮原子，和任选的1个选自下组的杂原子：O。
  3. 如权利要求1所述的N-苄基色胺酮类衍生物，其特征在于，所述的R2为取代或未取代的选自下组的基团：
    Figure PCTCN2014090947-appb-100002
    其中，
    Figure PCTCN2014090947-appb-100003
    表示连接位点；
    所述的取代指基团上的一个或多个氢原子被选自下组的取代基取代：C1-C4烷基、卤素。
  4. 一种N-苄基色胺酮类衍生物，其特征在于所述衍生物的结构通式如下：
    Figure PCTCN2014090947-appb-100004
    R1＝H,F
    Figure PCTCN2014090947-appb-100005
    其中：所述取代基R2为环状亚胺或二烷基取代胺，R1为氢或氟。
  5. 如权利要求1或4所述的N-苄基色胺酮类衍生物，其特征在于，所述衍生物为选自下组的化合物：
    Figure PCTCN2014090947-appb-100006
  6. 如权利要求1或4所述的N-苄基色胺酮类衍生物的制备方法，其特征在于，包括步骤：
    Figure PCTCN2014090947-appb-100007
    (d)在惰性溶剂中，在三乙胺存在下，用式5化合物(溴代2-甲基色胺酮)与R2H反应，得到式1化合物，
    式中，R1、R2的定义如上。
  7. 如权利要求6所述的方法，其特征在于，所述方法在步骤(d)之前还包括步骤(c)：
    Figure PCTCN2014090947-appb-100008
    (c)在惰性溶剂中，用式4化合物与溴代试剂反应，得到式5化合物；较佳地，所述的溴代试剂为NBS。
  8. 一种如权利要求4所述的N-苄基色胺酮类衍生物的制备方法，其特征在于，合成路线如下：
    Figure PCTCN2014090947-appb-100009
    R1＝H,F
    Figure PCTCN2014090947-appb-100010
    具体步骤如下：
    (1)甲基靛红酸酐(3)的合成
    5-甲基靛红悬浮于干燥二氯甲烷中，在0℃下分批加入间-氯过氧苯甲酸，在室温下搅拌1.5-2.5小时；TLC检测反应完全后，过滤反应形成的白色固体，用乙 酸乙酯洗涤三次，得到5-甲基靛红酸酐；其中，5-甲基靛红和间-氯过氧苯甲酸摩尔比为:(0.5-1):1；
    (2)2-甲基色胺酮(4)的合成
    将甲基靛红酸酐、5-氟靛红和三乙胺的混合物悬浮于干燥甲苯中，在100-120℃下加热3.5-4.5小时，溶剂在减压条件下蒸去，黄色的固体溶解于二氯甲烷中，再加入乙酸乙酯，形成的黄色固体经过滤用乙酸乙酯洗涤三次，得到2-甲基色胺酮；其中,甲基靛红酸酐和5-氟靛红和三乙胺摩尔比为:(0.2-0.5):(0.2-0.5):1；
    (3)溴代2-甲基色胺酮(5)的合成
    在75-85℃和氮气保护的条件下，将步骤(2)所得的2-甲基色胺酮溶解于干燥的二氯甲烷中，然后分批加入NBS和AIBN的混合物；反应液在75℃加热15-17小时，TLC检测反应完全；反应液冷却到室温后经盐水洗涤，无水硫酸钠干燥，浓缩得到的黄色产物；其中,2-甲基色胺酮和NBS和AIBN摩尔比为:1:1:(0.005-0.02)；
    (4)N-苄基色胺酮(1)的合成
    将溴代2-甲基色胺酮、脂肪胺和三乙胺在干燥DMF中于室温下搅拌1.5-2.5小时，TLC检测反应完全后，加入50 ml水，再用10 ml二氯甲烷萃取三次，合并的二氯甲烷相用水洗三次再用无水硫酸钠干燥；浓缩得到的黄色固体用硅胶分离，得到黄色的N-苄基色胺酮衍生物；其中,溴代2-甲基色胺酮和脂肪胺和三乙胺摩尔比为:1:(1-2):(2-5)。
  9. 一种如权利要求1或4所述的N-苄基色胺酮衍生物在制备预防和/或治疗具有IDO介导的色氨酸代谢紊乱的病理学特征的疾病的药物中的应用。
  10. 一种权利要求1或4所述的式1化合物或其药学上可接受的盐的用途，其特征在于，用于：
    (i)制备IDO抑制剂；
    (ii)制备色氨酸代谢紊乱相关疾病的药物；
    (iii)体外非治疗性地抑制IDO。
  11. 一种药物组合物，其特征在于，包含(i)如如权利要求1或4所述的式1化合物，或其药学上可接受的盐；以及(ii)药学上可接受的载体。
  12. 一种IDO抑制剂，其特征在于，包含如权利要求1或4所述的式1化合物，或其药学上可接受的盐。
  13. 一种体外非治疗性地抑制IDO活性的方法，其特征在于，所述方法包括步骤：使抑制有效量的如权利要求1或4所述的式1化合物，或其药学上可接受的盐与待抑制对象接触。
  14. 一种制备治疗色氨酸代谢紊乱相关疾病的药物组合物的方法，其特征在于，所述方法包括：将治疗有效量的如权利要求1或4所述的式1化合物或其药学上可接受的盐与药学上可接受的载体混合，从而形成药物组合物。
  15. 一种治疗色氨酸代谢紊乱相关疾病的方法，其特征在于，所述方法包括：对治疗对象施用治疗有效量的如权利要求1或4所述的式1化合物，或其药学上可接受的盐。
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US10059712B2 (en) 2013-11-12 2018-08-28 Fudan University N-benzyl tryptanthrin derivative, and preparation method and application thereof
CN110041335A (zh) * 2019-04-24 2019-07-23 同济大学 一种提高色胺酮的水溶性的方法、色胺酮衍生物及其制备方法和应用
CN110183454A (zh) * 2019-06-20 2019-08-30 同济大学 一种含1,2,3-三氮唑的色胺酮及其制备方法和应用
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CN110283192A (zh) * 2019-07-18 2019-09-27 同济大学 一种含硼酸的色胺酮衍生物的制备方法及应用
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US20160297822A1 (en) 2016-10-13
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