WO2018113624A1 - 具有吲哚胺2,3-双加氧酶抑制活性的稠合咪唑化合物 - Google Patents

具有吲哚胺2,3-双加氧酶抑制活性的稠合咪唑化合物 Download PDF

Info

Publication number
WO2018113624A1
WO2018113624A1 PCT/CN2017/116914 CN2017116914W WO2018113624A1 WO 2018113624 A1 WO2018113624 A1 WO 2018113624A1 CN 2017116914 W CN2017116914 W CN 2017116914W WO 2018113624 A1 WO2018113624 A1 WO 2018113624A1
Authority
WO
WIPO (PCT)
Prior art keywords
ethyl
imidazo
hydroxy
piperidin
isoindol
Prior art date
Application number
PCT/CN2017/116914
Other languages
English (en)
French (fr)
Inventor
余金迪
鲁先平
李志斌
辛利军
朱江飞
付超
Original Assignee
深圳微芯生物科技有限责任公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 深圳微芯生物科技有限责任公司 filed Critical 深圳微芯生物科技有限责任公司
Priority to EP17882589.9A priority Critical patent/EP3560928B1/en
Priority to US16/472,185 priority patent/US10604529B2/en
Publication of WO2018113624A1 publication Critical patent/WO2018113624A1/zh

Links

Classifications

    • 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
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/22—Anxiolytics
    • 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
    • A61P25/24—Antidepressants
    • 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
    • A61P35/00—Antineoplastic agents
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/91—Dibenzofurans; Hydrogenated dibenzofurans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6402—Atomic fluorescence; Laser induced fluorescence

Definitions

  • the invention relates to a fused imidazole compound having inhibitory activity of indoleamine 2,3-dioxygenase (IDO), a preparation method thereof and the same treatment with indoleamine-2,3-double Clinical application of diseases associated with abnormal oxygenase activity.
  • IDO indoleamine 2,3-dioxygenase
  • Trp Tryptophan
  • IDO Indoleamine 2,3-dioxygenase
  • Indoleamine 2,3-dioxygenase is an intracellular heme-containing enzyme that was first discovered in the intestine of rabbits in 1967 (Yamaoto S. et al., J Biol Chem, 1967, 242). (22): 5260-5266), the only rate-limiting enzyme outside the liver that catalyzes the epoxidation of purines in tryptophan molecules and catabolizes along the kynuric acid pathway (MacKenzie, CRet al. Current Drug Metabolism, 2007) , 8:237-244).
  • IDO is widely distributed in extrahepatic tissues, especially on the surface of mucosal tissues (such as placenta, lung and small intestine), thymic medulla and secondary lymphoid T cells, gastrointestinal mucosa, epididymis, placenta and anterior chamber of the anterior chamber.
  • mucosal tissues such as placenta, lung and small intestine
  • thymic medulla and secondary lymphoid T cells adenoid T cells
  • gastrointestinal mucosa epididymis
  • placenta and anterior chamber of the anterior chamber Epithelial cells, macrophages, dendritic cells (DCs), and microglia are less expressed in the spleen, lymph nodes, and thymus cortex (Fusao Hirta et al., J Biol Chem, 1997 252 ( 13): 4637-4642).
  • TDO is mainly expressed in the liver and controls the uptake of tryptophan to serotonin in food.
  • INF-specific inflammatory factors such as IFN-[gamma] stimulate the expression of IDO at the transcriptional level.
  • Other inflammatory factors such as IFN- ⁇ , IFN- ⁇ and LPS can also induce IDO expression, but the induction effect is not as good as that of IFN- ⁇ (King NJ et al., The Int J Biochem Cell Biol, 2007 39(12): 2167- 2172).
  • the expression of IDO is also regulated by immunologically active molecules such as prostaglandins, cell surface protein cytotoxic T lymphocyte-associated antigen (CTLA24), CD40, and Toll-like receptors.
  • CTL24 cell surface protein cytotoxic T lymphocyte-associated antigen
  • IDO is involved in the regulation of T cell regulation. IDO can cleave the activation of T cells by degrading tryptophan because T cells are particularly sensitive to tryptophan depletion. When the concentration of tryptophan is low, T cell proliferation will remain in G1 phase (Munn DH et al., J Exp Med, 1999, 189(9): 1363-1372). Based on this mechanism, IDO expressed in the placenta protects the fetus from maternal rejection (Munn DH et al., Science, 1998, 281 (5380): 1191-1193); whereas IDO expressed in tumors mediates tumors Immune escape (Friberg M.
  • IDOs in antigen-presenting cells such as macrophages and dendritic cells (DCs) can induce T cell immune tolerance to tumor antigens by inhibiting T cell proliferation (Terness P. et al., Blood, 2005, 105 ( 6): 2480-2486).
  • IDO is closely related to neurological diseases. IDO can affect the function of the brain through at least two mechanisms: 1) reducing the circulating tryptophan concentration by metabolizing tryptophan in the inflammatory reaction, thereby lowering the level of serotonin, leading to depression; 2) Catalytic tryptophan metabolism by kynurenine pathway accumulates kynurenine and neurotoxic quinolinic acid (Roy EJ et al., Neurosci Lett, 2005, 387(2): 95-99).
  • IDO 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 ultraviolet filter compounds increases with age (Takikawa et al. Adv. Exp. Med. Biol. 1999, 467, 241-245) and it has been reported that these ultraviolet filter compounds cause the lens to gradually become cloudy, which leads to the so-called Age-related nuclear cataract.
  • IDO inhibitors block this natural process (Takikawa O. et al. Exp. Eye Res. 2001, 72, 271-277).
  • IDO inhibitors can be used to treat or prevent diseases having pathological features of IDO-mediated tryptophan metabolism pathways, including infections such as AIDS, Lyme disease, and streptococcal infections.
  • IDO inhibitors there are currently three IDO inhibitors in different clinical stages, including: 1) Incyte's Epacadostat, in Phase II clinical trials for the treatment of myelodysplastic syndromes, melanoma and female reproductive system cancer; 2) Newlink's Indoximod, in clinical phase II, for the treatment of breast cancer, prostate cancer, malignant brain tumors, pancreatic cancer, and melanoma; 3) Roche's GDC-0919, in clinical phase I, for the treatment of advanced solid tumors.
  • IDO is closely related to a variety of disease pathogenesis, and has been confirmed to be a target for major diseases such as cancer, Alzheimer's disease, depression, cataract (CN101932325B, CN102579452B), so IDO inhibitors have broad application prospects as drugs.
  • major diseases such as cancer, Alzheimer's disease, depression, cataract (CN101932325B, CN102579452B)
  • IDO inhibitors have broad application prospects as drugs.
  • no suitable IDO inhibitors have been marketed so far, and IDO inhibitors currently in clinical research have the disadvantages of higher dosage and greater toxic side effects, so it is necessary to find IDO inhibition with higher IDO inhibitory activity and lower toxicity.
  • the agent has great theoretical significance and application value.
  • the compound of formula I according to the present invention can selectively inhibit indoleamine 2,3-dioxygenase, which is useful for the treatment and/or prevention of diseases having pathological features of IDO-mediated tryptophan metabolism pathway .
  • diseases include, but are not limited to, cancer, eye diseases, autoimmune diseases, psychological disorders, depression, and anxiety.
  • the invention provides a fused imidazole compound having indoleamine 2,3-dioxygenase inhibitory activity.
  • the invention provides a process for the preparation of a compound of the invention.
  • the compounds described herein include any suitable form thereof, including pharmaceutically acceptable salts, solvates, enantiomers thereof, and racemic mixtures thereof.
  • the invention further relates to a pharmaceutical composition
  • a pharmaceutical composition comprising as an active ingredient a compound according to the invention, including its various suitable forms.
  • the invention also provides the use of a compound of the invention in the manufacture of a medicament for use as a guanamine 2,3-dioxygenase (IDO) inhibitor.
  • the invention also provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prevention of a disease or condition such as cancer, ocular disease, autoimmune disease, psychological disorder, depression and anxiety.
  • the compound of the present invention has a chemical structure as shown in Formula I:
  • R 1 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 An alkylcarbonyl group and a C 1-6 alkylamino group;
  • R 2 is a hydroxyl group or an amino group
  • R 3 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcarbonyl And a C 1-6 alkylamino group;
  • X is NR 4 or O
  • R 4 is hydrogen or C 1-6 alkyl
  • Ring A is an optionally substituted group selected from the group consisting of phenyl; a 3- to 7-membered saturated or partially unsaturated carbocyclic ring; an 8- to 10-membered saturated, partially unsaturated or aromatic ring bicyclic or tricyclic ring; 1 to 4 5- to 6-membered monocyclic heteroaryl rings independently selected from nitrogen, oxygen or sulfur heteroatoms; having 4 to 3 heteroatoms independently selected from nitrogen, oxygen or sulfur to 4 a 7-membered saturated or partially unsaturated heterocyclic ring; a 7- to 10-membered saturated or partially unsaturated heterocyclic bicyclic or tricyclic ring having 1 to 5 hetero atoms independently selected from nitrogen, oxygen or sulfur; or 1 to 5 An 8- to 10-membered heteroaryl ring bicyclic or tricyclic ring independently selected from nitrogen, oxygen or sulfur heteroatoms.
  • R 1 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, and C 1-6 haloalkyl;
  • R 2 is a hydroxyl group
  • R 3 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, C 1-6 alkyl, C 1-6 alkoxy, and C 1-6 haloalkyl;
  • X is NR 4 ;
  • R 4 is hydrogen, methyl or ethyl
  • Ring A is a group selected from the group consisting of phenyl; a 3- to 6-membered saturated carbocyclic ring; an 8- to 10-membered aromatic ring bicyclic or tricyclic; having 1 to 4 independently selected from nitrogen, oxygen or sulfur.
  • a 5- to 6-membered monocyclic heteroaryl ring of an atom a 7- to 10-membered saturated or partially unsaturated heterocyclic bicyclic or tricyclic ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen or sulfur;
  • R 1 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro and halogen;
  • R 2 is a hydroxyl group
  • R 3 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, C 1-6 alkyl, C 1-6 alkoxy, and C 1-6 haloalkyl;
  • X is NR 4 ;
  • R 4 is hydrogen or methyl
  • Ring A is a group selected from the group consisting of phenyl; a 3- to 6-membered saturated carbocyclic ring; an 8- to 10-membered aromatic ring bicyclic or tricyclic; having 1 to 4 independently selected from nitrogen, oxygen or sulfur. a 5- to 6-membered monocyclic heteroaryl ring of an atom; or an 8- to 10-membered heteroaryl ring bicyclic or tricyclic ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen or sulfur.
  • R 1 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano and nitro;
  • R 2 is a hydroxyl group
  • R 3 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, C 1-6 alkyl, C 1-6 alkoxy, and C 1-6 haloalkyl;
  • X is NR 4 ;
  • R 4 is hydrogen or methyl
  • Ring A is a group selected from the group consisting of phenyl; 8- to 10-membered aromatic ring bicyclic or tricyclic; 5- to 6-membered single ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur. a heteroaryl ring; or an 8- to 10-membered heteroaryl ring bicyclic or tricyclic having 1 to 5 heteroatoms independently selected from nitrogen, oxygen or sulfur.
  • R 1 is one or more substituents selected from hydrogen, hydroxy or cyano
  • R 2 is a hydroxyl group
  • R 3 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, C 1-6 alkyl, C 1-6 alkoxy, and C 1-6 haloalkyl;
  • X is NR 4 ;
  • R 4 is hydrogen or methyl
  • Ring A is a group selected from the group consisting of phenyl; a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur; or having 1 to 5 independent
  • R 1 is hydrogen
  • R 2 is a hydroxyl group
  • R 3 is one or more substituents selected from the group consisting of hydrogen, hydroxy, cyano, nitro, halogen, C 1-6 alkyl, C 1-6 alkoxy, and C 1-6 haloalkyl;
  • X is NR 4 ;
  • R 4 is hydrogen or methyl
  • Ring A is a group selected from the group consisting of phenyl; a 5- to 6-membered monocyclic heteroaryl ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur; or having 1 to 5 independent
  • halogen described in the present invention is fluorine, chlorine, bromine or iodine.
  • alkyl refers to having from 1 to about 20 carbon atoms, usually from 1 to 12 carbon atoms, preferably from 1 to 8, from 1 to 6, even more preferably from 1 to 4 carbons. Linear and branched or cyclic alkyl groups of atoms.
  • linear alkyl group examples include those having 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl and n-octyl; branched alkyl groups Examples include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, isohexyl, and 2,2-dimethylpropyl; examples of cycloalkyl include It is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
  • haloalkyl group as used in the present invention includes a monohaloalkyl group and a polyhalogenated alkyl group (wherein all halogen atoms may be the same or different).
  • Partially halogenated alkyl is a "haloalkyl" group as used herein.
  • the haloalkyl group include a trifluoromethyl group, a 1,1-dichloroethyl group, a 1,2-dichloroethyl group, a 1,3-dibromo-3,3-difluoropropyl group and the like.
  • alkoxy group refers to a group formed by linking the above alkyl group to an oxygen atom, wherein the oxygen atom has a free bonding ability such as a methoxy group, an ethoxy group, a propoxy group, or a butyl group.
  • “Pharmaceutically acceptable salt” or “pharmaceutically acceptable salt” as used in the present invention refers to pharmaceutically acceptable acid and base addition salts and solvates.
  • Such pharmaceutically acceptable salts include those formed with acids including hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfinic acid, formic acid, p-toluenesulfonic acid, methanesulfonic acid, nitric acid, benzoic acid, lemon.
  • substituted means that an organic group as defined herein, which contains one or more bonds bonded to a hydrogen atom, is substituted by one or more bonds bonded to a non-hydrogen atom or a radical.
  • the non-hydrogen atom or atomic group is a substituent.
  • the substituent of the compound of the present invention includes, for example, a halogen, an alkyl group (preferably a C 1-8 alkyl group, a C 1-6 alkyl group or a C 1-4 alkyl group), an alkoxy group (preferably a C 1-8 alkoxy group, C).
  • haloalkyl preferably C 1-8 haloalkyl, C 1-6 haloalkyl or C 1-4 haloalkyl
  • haloalkoxy preferably C 1- 8 haloalkoxy, C 1-6 haloalkoxy or C 1-4 haloalkoxy
  • hydroxy preferably hydroxy C 1-8 alkyl, hydroxy C 1-6 alkyl or hydroxy C 1-4 alkyl.
  • the invention relates to fluorescent, spin-labeled, heavy metal or isotopically-labeled derivatives of the compounds described herein, which are useful not only for imaging, but also for in vivo and in vitro detection, localization and quantification of tissue samples (including humans)
  • the IDO enzyme the IDO enzyme ligand is recognized by inhibition of binding of the labeled compound.
  • the present invention further provides an IDO enzyme detection reagent or kit containing such a labeled compound.
  • the invention further provides isotopically labeled compounds of the compounds of the invention.
  • An “isotopically labeled compound” or “isotopically labeled” compound of the invention as used herein refers to a compound described herein in which one or more atoms are replaced by an isotope atom having an atomic mass or mass different from the atomic mass. Usually found in nature (ie naturally occurring Atomic mass or mass number. Suitable radionuclides may include, but are not limited to, 2H ( ⁇ , also written as D), 3H ( ⁇ .
  • radioisotope contained in an isotopically labeled compound will depend on the particular application of the isotopically labeled compound. For example, for labeling and competition assays of IDO enzymes in vitro, compounds comprising 3H, 14C, 82Br, 125I, 131I, 35S are generally most useful. For isotope imaging applications, 11C, 18F, 125I, 123I, 124I, 131I, 75Br, 76Br or 77Br are generally most useful.
  • the invention provides a process for the preparation of a compound of formula I of the invention:
  • the method comprises formulating a compound of formula C
  • R 1 , R 2 , R 3 , X and A are as defined above.
  • the invention also provides a process for the preparation of a compound of formula C:
  • the compound of formula C is formed by coupling with an organic solvent and a coupling reagent, wherein R 1 , R 3 , X and A are as defined above.
  • the invention also provides the use of a compound of formula C as an intermediate for the preparation of a compound of formula I according to the invention.
  • the present invention further provides a process for the preparation of a compound of the above formula A, comprising the following steps (a)-(d):
  • compound 5 is removed from the trityl group under acetic acid conditions and is ring-closed to give compound 6;
  • the above-mentioned Suzuki cross-coupling reaction (a) uses palladium as a catalyst, such as Pd(PPh 3 ) 4 , Pd(dffp)Cl 2 , Pd(OAc) 2 , Pd(dba) 3 /PCy 3 , PdCl 2 , Pd (PPh 3 ) 4 Cl 2 and the like.
  • Ni as a catalyst, such as NiCl 2 (dffp), NiCl 2 (dffp) / Zn, NiCl 2 (dffp) / BuLi, NiCl 2 (PPh 3 ) 2 / PPh 3 , NiCl 2 (NEt 3 ) 2 , NiCl 2 (NEt 3 ) 2 , NiCl(bipy), Ni(TPPS) 3 , Ni(COD) 2 , NiCl 2 (PPh 3 ) 2 /n-BuLi, Ni ⁇ P(OMe) 3 ⁇ 2 Cl 2 , NiCl 2 ( PCy 3 ) 2 and so on.
  • KOAc, K 3 PO 4 , K 2 CO 3 , NaOH, Ba(OH) 2 , Na 2 CO 3 , CsF or NaHCO 3 is used, the reaction temperature is 25 to 140 ° C, and the reaction time is 4 to 72 hours.
  • the solvent used in the reaction is a usual solvent such as ethanol, THF, isopropanol, DMSO, DMF, dioxane, toluene, water, and DME.
  • the above-mentioned Claisen-Schimidt condensation reaction (b) may be carried out using a base such as NaOH, NaOCH 3 or NaOEt as a catalyst, or an acid as a catalyst such as sulfuric acid, hydrochloric acid, acetic acid or the like.
  • the reaction temperature is 0 to 80 ° C, and the reaction time is 1 to 24 hours.
  • the solvent used for the reaction may be a usual solvent such as THF, ethanol, water or the like.
  • the above reaction (c) uses acetic acid, hydrochloric acid and trifluoroacetic acid as deprotection and ring-closing reagents, the reaction temperature is 0 to 90 ° C, and the reaction time is 2 to 72 hours.
  • the solvent used for the reaction is a usual solvent such as water, methanol, ethanol, isopropanol, tetrahydrofuran, dichloromethane, acetone or the like.
  • the above deprotection reaction (d) is hydrochloric acid, trifluoroacetic acid, p-toluenesulfonic acid, sulfuric acid, methanesulfonic acid, boron trifluoride diethyl ether or the like as a deprotecting reagent, the reaction temperature is 25 to 100 ° C, and the reaction time is 20 minutes. 24 hours.
  • the solvent used in the reaction is a usual solvent such as water, methanol, ethanol, ethyl acetate, dichloromethane, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide or the like.
  • X is O; X is NR 4 .
  • the nucleophilic substitution reaction is NaOH, Na2CO 3, K 2 CO 3 as base to acid and the like, the reaction temperature is 25 ⁇ 140 °C, the reaction time is 2 to 72 hours.
  • the solvent used for the reaction is a usual solvent such as water, methanol, ethanol, acetonitrile, benzene, xylene, acetone, N,N'-dimethylformamide, DMSO or the like.
  • the obtained compound 9 is deethylated under the action of a base such as NaOH or LiOH to obtain a compound B (1).
  • the reaction temperature is 0 to 60 ° C, and the reaction time is 0.5 to 2 hours.
  • Commonly used solvents for the reaction are water, methanol, ethanol, tetrahydrofuran, N,N'-dimethylformamide and the like.
  • the compound of formula B(2) can be purchased directly from the market or prepared by the following synthetic methods:
  • a commercially available amine 10 is nucleophilicly substituted with a commercially available compound 8 under basic conditions to give compound 11.
  • the nucleophilic substitution reaction is NaOH, Na 2 CO 3, K2CO 3 , etc. as a base to the acid, the reaction temperature is 25 ⁇ 140 °C, the reaction time is 2 to 72 hours.
  • the solvent used for the reaction is a usual solvent such as water, methanol, ethanol, acetonitrile, benzene, tetrahydrofuran, xylene, acetone, N,N'-dimethylformamide, DMSO or the like.
  • the obtained compound 11 is deethylated under the action of a base such as NaOH or LiOH to give a compound of the formula B (2).
  • the reaction temperature is 0 to 60 ° C, and the reaction time is 0.5 to 2 hours.
  • Commonly used solvents for the reaction are water, methanol, ethanol, tetrahydrofuran, N,N'-dimethylformamide and the like.
  • the compounds of formula I according to the invention may be prepared by the following synthetic methods:
  • the compound of formula A is condensed with a compound of formula B to give a compound of formula C.
  • the condensation reaction uses a peptide condensing agent as a catalyst, such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), N,N'-dicyclohexylcarbodiimide (DCC). , N, N'-carbonyldiimidazole (CDI), O-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and the like.
  • the reaction temperature is 0 to 60 ° C, and the reaction time is 2 to 72 hours.
  • the solvent used for the reaction is a usual solvent such as benzene, toluene, tetrahydrofuran, dioxane, dichloromethane, chloroform, N,N'-dimethylformamide or the like. If necessary, a base such as sodium hydroxide, triethylamine or pyridine may also be added.
  • a reducing agent such as NaBH 4 , KBH 4 , NaBH(OAc) 3 , KBH(OAc) 3 , NaBH 3 CN or the like.
  • the reaction temperature is 0 to 60 ° C and the reaction time is 2 to 72 hours.
  • the solvent used in the reaction is a usual solvent such as methanol, ethanol, tetrahydrofuran or the like.
  • the reaction temperature is 0 to 25 ° C, and the reaction time is 0.5 to 16 hours.
  • the solvent used in the reaction is a usual solvent such as tetrahydrofuran, diethyl ether, dichloromethane, toluene, ethyl acetate, acetonitrile, N,N'-dimethylformamide or the like.
  • the compound of the formula (I) can be purified by a usual separation method such as extraction, recrystallization, column chromatography and the like.
  • 4-iodo-1H-imidazole (20 g, 103 mmoles) was dissolved in DMF (100 mL) then triethylamine (15.1 mL, 108 mmoles) and triphenylmethyl chloride (27.8 g, 100 mmoles). The reaction mixture was stirred at room temperature for 48 h, then poured into ice water (500 mL), and a solid solid was precipitated, which was filtered and dried to give 4-ethyl-l-triphenylmethyl-1H-imidazole (40 g, 91.7 mmol, 88.9%).
  • 6-Chloro-9H-indole (3.08 g, 20.0 mmol) and DABCO (6.72 g, 60.0 mmol) were dissolved in DMSO (30 mL), stirred at 20 ° C for 4 h, then warmed to 35 ° C for 0.5 h.
  • DMSO DMSO
  • 2-hydroxyacetic acid ethyl ester (20.8 g, 200 mmol) NaH (6.40 g, 160 mmol) and DMSO (30 mL).
  • the reaction mixture was stirred at rt EtOAc (EtOAc)EtOAc.
  • the organic phase was dried over anhydrous sodium sulfate (MgSO4) , 0.450 mmol, 2.25%).
  • 4-(1-Hydroxy-2-(5H-imidazo[5,1-a])isoindol-5-yl)ethyl)piperidine-1-carboxylic acid tert-butyl ester is obtained from 4-(2-( 5H-Imidazo[5,1-a])isoindol-5-yl)acetyl)piperidine-1-carboxylic acid tert-butyl ester was prepared according to a similar procedure in Example 11.
  • Ethyl-1-one (90 mg, crude) is from 2-(3-trifluoromethylphenoxy)acetic acid (41.6 mg, 0.189 mmol) and 2-(5H-imidazo[5,1-a ]) Isoindole-5-yl)-1-(piperidin-4-yl)ethyl-1-one hydrochloride (50 mg, 0.158 mmol).
  • N-(3-Chloro-4-fluorophenyl)-N-methylglycine (170 mg, 95%) was obtained from N-(3-chloro-4-fluorophenyl)-N-methylglycine ethyl ester (200 mg, 0.816 mmol) was prepared according to a similar procedure in Example 61.
  • Yellow oil N-(4-fluorophenyl)-N-methylglycine ethyl ester (0.8 g, 95%) was obtained from 4-fluoro-N-methylaniline (500 mg, 4 mmol) and ethyl 2-bromoacetate (1.33 g, 8 mmol) was prepared according to a similar procedure as in Example 60.
  • N-(4-fluorophenyl)-N-methylglycine (600 mg, 98%) was obtained from N-(4-fluorophenyl)-N-methylglycine ethyl ester (700 mg, 3.3 mmol)
  • a similar procedure was carried out in Example 61.
  • N-Ethyl-N-phenylglycine ethyl ester (3.2 g, 85%) was obtained from N-ethylaniline (2.0 g, 16.5 mmol) and ethyl 2-bromoacetate (4.11 g, 24.8 mmol). A similar step in 60 was prepared.
  • N-ethyl-N-phenylglycine (90 mg, crude) was obtained from N-ethyl-N-phenylglycine ethyl ester (100 mg, <RTIgt;
  • Brown solid 1-(4-(2-(5H-imidazo[5,1-a]isoindol-5-yl)acetyl)piperidin-1-yl)-2-(ethyl(phenyl) Amino)ethyl-1-one 70 mg, crude
  • Phenylglycine (1.51 g, 10 mmol), (Boc) 2 O (3.27 g, 15 mmol) and sodium carbonate (2.12 g, 20 mmol) were dissolved in methanol (50 mL) and water (20 mmol). , 10 mmol), the pH was between 9 and 10, and allowed to react at room temperature overnight.
  • reaction mixture was diluted with water (40 mL), and the mixture was adjusted to pH 1-2 with hydrochloric acid, extracted with dichloromethane (40 mL*2), washed with water (20mL*2), and the organic phase was concentrated under reduced pressure to obtain a tan liquid N -(tert-Butoxycarbonyl)-N-phenylglycine (2.49 g, yield 98%).
  • N-methyl-N-(4-trifluoromethylphenyl)glycine ethyl ester (630 mg, 84%) was obtained from N-methyl-4-trifluoromethylaniline (500 mg, 2.86 mmol)
  • Ethyl 2-bromoacetate (0.96 g, 5.72 mmol) was prepared according to a similar procedure from Example 60.
  • N-methyl-N-(4-trifluoromethylphenyl)glycine (470 mg, 87%) was obtained from N-methyl-N-(4-trifluoromethylphenyl)glycine ethyl ester (600 mg, 2.3 mmol) was prepared according to a similar procedure in Example 61.
  • Yellow oil 1-(4-(2-(5H-imidazo[5,1-a]isoindol-5-yl)acetyl)piperidin-1-yl)-2-(methyl (4- Trifluoromethylphenyl)amino)ethyl-1-one (60 mg, 26%) consists of N-methyl-N-(4-trifluoromethylphenyl)glycine (140 mg, 0.6 mmol) and 2- (5H-Imidazo[5,1-a])isoindole-5-yl)-1-(piperidin-4-yl)ethyl-1-one hydrochloride (130 mg, 0.46 mmol).
  • a similar step in 62 was prepared.
  • N-(4-cyanophenyl)-N-methylglycine 250 mg, 92%) was obtained from N-(4-carbonitrile-phenyl)-N-methylglycine ethyl ester (310 mg, 1.42 mmol) Prepared according to a similar procedure as in Example 61.
  • N-(3-Methoxyphenyl)-N-methylglycine ethyl ester 400 mg, 49%) was obtained from 3-methoxy-N-methylaniline (500 mg, 3.65 mmol) and 2-bromoacetic acid
  • the ester (731 mg, 4.38 mmol) was prepared in a similar procedure as in Example 60.
  • N-(3-Methoxyphenyl)-N-methylglycine 300 mg, 85% was obtained from N-(3-methoxyphenyl)-N-methylglycine ethyl ester (400 mg, 1.8 mmol) Prepared according to a similar procedure as in Example 61.
  • Yellow oil 1-(4-(2-(5H-imidazo[5,1-a]isoindol-5-yl)acetyl)piperidin-1-yl)-2-(methyl-pyridine- 2-yl)amino)ethyl-1-one 70 mg, 29%) consists of N-methyl-N-(pyridin-2-yl)glycine (122 mg, 0.74 mmol) and 2-(5H-imidazo[ 5,1-a])isoindole-5-yl)-1-(piperidin-4-yl)ethyl-1-one hydrochloride (160 mg, 0.57 mmol). Made.
  • Ethylquinoline-6-ylglycine ethyl ester (1200 mg, 5.21 mmol) was dissolved in methanol (20 ml), and formaldehyde (1513 mg, 52.1 mmol) was added thereto, and the reaction mixture was stirred at room temperature for 2 hours, and then cyano boron was added thereto.
  • Sodium hydride (1643 mg, 26.1 mmol). The mixture was stirred at room temperature for 16 hours. 50 ml of water was added to the reaction solution, and extracted with ethyl acetate (30 ml).
  • the combined organic phases were dried with anhydrous sodium s
  • Yellow oil 1-(4-(2-(5H-imidazo[5,1-a]isoindol-5-yl)acetyl)piperidin-1-yl)-2-(methyl(quinoline) -6-yl)amino)ethyl-1-one (45 mg, 26%) consists of N-methyl-N-(quinolin-6-yl)glycine (100 mg, 0.46 mmol) and 2-(5H-imidazole And [5,1-a])isoindole-5-yl)-1-(piperidin-4-yl)ethyl-1-one hydrochloride (100 mg, 0.36 mmol). The steps are prepared.
  • N-Methyl-N-(9-(tetrahydro-2H-pyran-2-yl)-9H-indol-6-yl)glycine ethyl ester (550 mg, 1.72 mmol) was dissolved in methanol (10 ml) and tetrahydrofuran ( 10 ml), adding 10 ml of lithium hydroxide (1 mol / liter), the mixture was reacted at room temperature for 3 hours, the reaction was stopped, the reaction solution was adjusted to pH 6 with 1 mol / liter of hydrochloric acid, and extracted with ethyl acetate 3 times, combined organic The layer was washed three times with saturated brine, dried over anhydrous sodium sulfate -yl)glycine (430 mg, 86%).
  • N-methyl-N-(9-(tetrahydro-2H-pyran-2-yl)-9H-indol-6-yl)glycine (170 mg, 0.58 mmol) dissolved in hydrochloric acid / dioxane solution (10 ml) and DMF (0.5 ml). The reaction solution was stirred at room temperature for 1 hour. Dry to give N-methyl-N-(9H-purin-6-yl)glycine as a yellow oil (130 mg, 92%).
  • Yellow oil 1-(4-(2-(5H-imidazo[5,1-a]isoindol-5-yl)acetyl)piperidin-1-yl)-2-(methyl (7H-) Indole-6-yl)amino)ethyl-1-one (60 mg, 24%) consists of N-methyl-N-(9H-indol-6-yl)glycine (130 mg, 0.69 mmol) and 2-(5H -Imidazo[5,1-a])isoindol-5-yl)-1-(piperidin-4-yl)ethyl-1-one hydrochloride (150 mg, 0.54 mmol) A similar step was prepared.
  • Yellow oil dibenzo[b,d]furan-3-ylglycine ethyl ester (570 mg, 65%) from 3-aminodibenzo[b,d]furan (600 mg, 3.3 mmol) and 2-bromoacetic acid Ethyl ester (821 mg, 4.9 mmol) was prepared according to a similar procedure from Example 64.
  • N-(dibenzo[b,d]furan-3-yl)-N-methylglycine ethyl ester (350 mg, 83%) was obtained from dibenzo[b,d]furan-3-ylglycine
  • the ester 400 mg, 1.48 mmol
  • iodomethane (1.05 g, 7.43 mmol) were prepared in a similar procedure as in Example 65.
  • the assay is used for in vitro biological activity evaluation of the compounds of the invention, including in vitro enzymatic activity evaluation and cell level enzymatic activity evaluation.
  • This assay was to comprehensively evaluate the in vitro enzymatic inhibitory activity of different compounds on human indoleamine 2,3-dioxygenase (IDO) and the enzymatic inhibitory activity on cell models.
  • the basic principle of in vitro IDO enzymatic activity detection is to use the IDO enzyme to metabolize the substrate L-tryptophan to produce the product kynurenine in an in vitro enzyme-catalyzed reaction system.
  • the latter can be combined with p-dimethylamino.
  • the glacial acetic acid solution of benzaldehyde was yellow in color and the absorbance was measured at 492 nm.
  • the concentration of the product kynurenine in the sample was calculated by comparison to a known concentration of the kynurenine standard curve. When different test compounds were added, their inhibition of IDO enzyme activity was manifested by a decrease in the product kynurenine and a change in color response.
  • IDO1 enzyme Recombinant human IDO enzyme
  • Detection reagents such as L-tryptophan (Biotech, A601911-0050), ascorbic acid (production, SB0830-100g), catalase (production, 54J15066), methylene blue (Tianjin Benchmark Chemical Co., Ltd.), dog urine Amino acid (sigma, K8625-100MG), trichloroacetic acid (raw labor, A600968-0250), p-dimethylaminobenzaldehyde (Tianjin Damao Chemical Reagent Factory).
  • the detection reaction was simultaneously provided with a control reaction including a 0 inhibition positive control to which no test compound was added and a 0 enzyme negative control to which no enzyme was added. Multiple holes were used for all tests.
  • the detection wavelength is 492nm.
  • the compound of the present invention was subjected to in vitro IDO enzymatic evaluation under the same conditions using the compound NLG919 (CAS: 1402836-58-1) known in the prior art as a positive control compound (for IDO1 detection, The test compound concentration was 200 nM).
  • the data summary is shown in the table below (Table 1).
  • the compounds of the present invention have good IDO enzyme inhibitory activity, and most of them are superior to the control compound NLG919.
  • the compound of the present invention was subjected to an in vitro IDO enzymology IC 50 measurement using NLG919 as a positive control compound.
  • the data summary is shown in the table below (Table 2), and it can be seen that the representative compounds of the present invention have lower in vitro IDOIC 50 values.
  • IDO enzymes in addition to constitutive expression of IDO enzymes in immune cells such as myeloid-derived suppressor cells (MDSCs), many tumor cells also up-regulate expression of IDO, or are induced by cytokines such as IFN- ⁇ .
  • IFN- ⁇ we used IFN- ⁇ to induce Hela cells to express the IDO enzyme (IDO1 enzyme) as a model to detect the IDO enzymatic inhibitory activity of the compound at the cellular level.
  • HeLa cells are human cervical cancer cell lines that up-regulate the expression of endogenous IDO enzymes under the induction of human IFN- ⁇ .
  • the cell supernatant can be subjected to detection of the enzyme-catalyzed product kynurenine by adding the substrate L-tryptophan to the cell culture solution.
  • the cultured Hela cells were cultured under the stimulation of human IFN- ⁇ , and after incubation with different concentrations of test compounds for a specified time, the color reaction of the enzyme product with p-dimethylaminobenzaldehyde was used to detect the test compound treatment on the cell IDO enzyme. The effect of activity.
  • Recombinant human IFN-[gamma] cytokine was purchased from Biotech, Inc. Cell culture was purchased from Gibco with phenol red free DMEM. Detection reagents such as L-tryptophan (Biotech, A601911-0050), kynurenine (sigma, K8625-100MG), trichloroacetic acid (production, A600968-0250), p-dimethylaminobenzaldehyde (Tianjin Da Mao Chemical Reagent Factory), 96-well flat bottom plate for cell culture (CORNING, costar 3599).
  • Hela cells were inoculated into 96-well culture plates at an appropriate concentration (about 20,000 cells/well), and after night-stacking, they were replaced with phenol red-free DMEM medium containing 200 ⁇ M L-tryptophan.
  • the compound of the present invention was subjected to cytological level IDO enzymatic evaluation (test compound concentration: 1 ⁇ M) using NLG919 as a positive control compound.
  • the data summary is shown in the table below (Table 3).
  • Table 3 the results in Table 3, the compounds of the present invention have good IDO inhibitory activity, and most are superior to the control compound NLG919.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Neurosurgery (AREA)
  • Neurology (AREA)
  • Biomedical Technology (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Optics & Photonics (AREA)
  • Pathology (AREA)
  • Pain & Pain Management (AREA)
  • Psychiatry (AREA)
  • Ophthalmology & Optometry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

公开了一类稠合咪唑化合物、其制备方法及其应用。所述化合物的结构如通式I 所示，其中各基团的定义如说明书所述。这些化合物能够选择性地抑制吲哚胺2,3-双加氧酶(IDO)。所述化合物可作为IDO抑制剂用于治疗和/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，例如癌症、眼部疾病、自身免疫性疾病、心理障碍、抑郁症、焦虑症及其它疾病。

Description

具有吲哚胺2,3-双加氧酶抑制活性的稠合咪唑化合物
本申请要求于2016年12月20日提交中国专利局、申请号为201611186194.4、发明名称为“具有吲哚胺2,3-双加氧酶抑制活性的稠合咪唑化合物”的专利申请，其全部内容通过引用结合在本申请中。
技术领域
本发明涉及具有吲哚胺2,3-双加氧酶(Indoleamine 2,3-dioxygenase,IDO)抑制活性的稠合咪唑化合物、其制备方法及其在治疗与吲哚胺-2,3-双加氧酶活性异常相关的疾病方面的临床应用。
背景技术
色氨酸(Trp)是人体必需的一种氨基酸。从饮食中获得的色氨酸，一部分用来合成蛋白质、烟酸以及神经递质5-羟色胺，其余部分则主要通过犬尿氨酸途径(kynurenine pathway)进行代谢(Leklem J.E.,Am J Clin Nutr,1971,24(6):659-672)。吲哚胺2,3-双加氧酶(Indoleamine 2,3-dioxygenase,IDO)是参与这种代谢途径的关键酶。
吲哚胺2,3-双加氧酶是一种细胞内含亚铁血红素的酶,于1967年首次在兔的肠道中被发现(Yamaoto S.et al.,J Biol Chem,1967,242(22):5260-5266)，是肝脏以外唯一可催化色氨酸分子中吲哚环氧化裂解、沿犬尿酸途径进行分解代谢的限速酶(MacKenzie,C.R.et al.Current Drug Metabolism,2007,8:237-244)。
IDO广泛分布于肝外组织，特别是粘膜组织表面(如胎盘、肺和小肠)、胸腺髓质和次级淋巴器官T细胞区、胃肠道粘膜、附睾、胎盘及眼前房等成纤维细胞、上皮细胞、巨噬细胞、树突状细胞(Dendritic cells,DC)以及小胶质细胞,而在脾脏、淋巴结和胸腺皮质中的表达较低(Fusao Hirta et al.,J Biol Chem,1997 252(13):4637-4642)。
TDO主要表达在肝脏中，并且控制食物中摄取的色氨酸向血清素和 犬尿氨酸路径的流量。
INF特异性炎性因子如IFN-γ在转录水平刺激诱导IDO的表达。其他炎性因子，如IFN-α，IFN-β及LPS也可诱导IDO表达，但诱导效果不如IFN-γ(King N.J.et al.,The Int J Biochem Cell Biol,2007 39(12):2167-2172)。同时IDO的表达还受到免疫活性分子如前列腺素，细胞表面蛋白细胞毒性T淋巴细胞相关抗原(CTLA24)、CD40、Toll样受体的调节。
最近的研究表明，IDO参与调节T细胞的调节。IDO通过降解色氨酸可切断T细胞的活化，因为T细胞对色氨酸耗竭特别敏感,在色氨酸浓度较低时,T细胞增殖就会静止在G1期(Munn D.H.et al.,J Exp Med,1999,189(9):1363-1372)。基于这种机制，在胎盘中表达的IDO保护了胎儿免遭母体排斥(Munn D.H.et al.,Science,1998,281(5380):1191-1193)；而在肿瘤中表达的IDO介导了肿瘤的免疫逃逸(Friberg M.et al.,Int J Cancer,2002,101(2):151-155)。抗原呈递细胞如巨噬细胞、树突状细胞(DC)上的IDO均可通过抑制T细胞增殖来诱导T细胞对肿瘤抗原的免疫耐受(Terness P.et al.,Blood,2005,105(6):2480-2486)。
IDO与神经系统疾病密切相关，IDO至少可通过两种机制影响脑的功能:1)在炎症反应时通过代谢色氨酸降低循环的色氨酸浓度，从而使5-羟色胺水平降低，导致抑郁；2)催化色氨酸循犬尿氨酸途径代谢使犬尿氨酸和神经毒性喹啉酸累积(Roy E.J.et al.,Neurosci Lett,2005,387(2):95-99)。
IDO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(Takikawa et al.Adv.Exp.Med.Biol.1999,467,241-245)并且已经报道了这些紫外线滤器化合物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抑制剂会阻断该自然过程(Takikawa O.et al.Exp.Eye Res.2001,72,271-277)。
现有技术已知，IDO抑制剂可以用于治疗或预防具有IDO介导的色氨酸代谢途径的病理学特征的疾病，这些疾病包括例如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(CN1795187A，CN101932325A，CN103054870A)。
目前已有三个IDO抑制剂处于不同的临床研究阶段，包括：1)Incyte公司的Epacadostat，处于临床II期，用于治疗骨髓增生异常综合症、黑色素瘤和女性生殖系统癌症；2)Newlink公司的Indoximod，处于临床II期，用于治疗乳腺癌、前列腺癌、恶性脑瘤、胰腺癌和黑色素瘤；3)罗氏公司的GDC-0919，处于临床I期，用于治疗晚期实体瘤。
IDO与多种疾病发病机制密切相关，并已被证实是癌症、阿尔茨海默病、抑郁症、白内障等重大疾病的靶标(CN101932325B，CN102579452B)，因此IDO抑制剂作为药物具有广阔的应用前景。然而，迄今未有合适的IDO抑制剂上市，现有处于临床研究阶段的IDO抑制剂具有使用剂量较高，毒副作用较大的缺点，因此寻找具有更高IDO抑制活性、更低毒性的IDO抑制剂具有重大的理论意义和应用价值。
发明内容
为了克服现有技术中的缺陷，本发明人进行了大量研究，通过大量筛选试验后意外发现，具有以下通式I的化合物具有出乎意料地好的IDO抑制作用：
Figure PCTCN2017116914-appb-000001
本发明所述的通式I化合物，可以选择性地抑制吲哚胺2,3-双加氧酶，可用于治疗和/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。这样的疾病包括但不限于癌症、眼部疾病、自身免疫性疾病、心理障碍、抑郁症和焦虑症。
一方面，本发明提供了具有吲哚胺2,3-双加氧酶抑制活性的稠合咪唑化合物。
另一方面，本发明目提供了制备本发明所述化合物的方法。
本发明所述的化合物包括其任何适当的形式，包括其药学上可接受的盐、溶剂化物、对映体以及外消旋体混合物。
本发明还涉及包含本发明所述化合物(包括其各种适当形式)作为活性成分的药物组合物。
在另一方面，本发明还提供了本发明化合物在制备用作吲哚胺2,3-双加氧酶(IDO)抑制剂的药物中的应用。在又一方面，本发明还提供了本发明化合物在制备用于治疗或预防癌症、眼部疾病、自身免疫性疾病、心理障碍、抑郁症和焦虑症等疾病或病症的药物中的应用。
本发明所说的化合物，其化学结构如通式I所示：
Figure PCTCN2017116914-appb-000002
其中，
R1为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基、C1-6卤代烷基、C1-6烷基羰基和C1-6烷基氨基；
R2为羟基或氨基；
R3为一个或多个取代基，选自氢、羟基、氰基、卤素、C1-6烷基、C1-6烷氧基、C1-6卤代烷基、C1-6烷基羰基和C1-6烷基氨基；
X为NR4或O；
其中，R4为氢或C1-6烷基；
环A为任选经取代的选自以下的基团：苯基；3元到7元饱和或部分不饱和碳环；8元到10元饱和、部分不饱和或芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；具有1到3个独立地选自氮、氧或硫的杂原子的4元到7元饱和或部分不饱和杂环；具有1到5个独立地选自氮、氧或硫的杂原子的7元到10元饱和或部分不饱和杂环双环或三环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳环双环或三环。
对于上述通式I所示化合物，优选地，
R1为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素和C1-6卤代烷基；
R2为羟基；
R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
X为NR4；
其中，R4为氢、甲基或乙基；
环A为选自以下的基团：苯基；3元到6元饱和碳环；8元到10元芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；具有1到5个独立地选自氮、氧或硫的杂原子的7元到10元饱和或部分不饱和杂环双环或三环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳环双环或三环。
对于上述通式I所示化合物，较优选地，
R1为一个或多个取代基，选自氢、羟基、氰基、硝基和卤素；
R2为羟基；
R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
X为NR4；
其中，R4为氢或甲基；
环A为选自以下的基团：苯基；3元到6元饱和碳环；8元到10元芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
对于上述通式I所示化合物，更优选地，
R1为一个或多个取代基，选自氢、羟基、氰基和硝基；
R2为羟基；
R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
X为NR4；
其中，R4为氢或甲基；
环A为选自以下的基团：苯基；8元到10元芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
对于上述通式(I)所示化合物，还更优选地，
R1为一个或多个取代基，选自氢、羟基或氰基；
R2为羟基；
R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6 烷基、C1-6烷氧基和C1-6卤代烷基；
X为NR4；
其中，R4为氢或甲基；
环A为选自以下的基团：苯基；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
对于上述通式(I)所示化合物，最优选地，
R1为氢；
R2为羟基；
R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
X为NR4；
其中，R4为氢或甲基；
环A为选自以下的基团：苯基；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
本发明所述的“卤素”为氟、氯、溴、碘。
本发明所述的“烷基”，是指具有1至约20个碳原子、通常为1至12个碳原子、优选为1至8个、1至6个甚至更优选为1至4个碳原子的直链和支链或环状烷基。直链烷基的实例包括具有1至8个碳原子的那些，比如甲基、乙基、正丙基、正丁基、正戊基、正己基、正庚基和正辛基；支链烷基的实例包括但不限于异丙基、异丁基、仲丁基、叔丁基、新戊基、异戊基、异己基和2,2-二甲基丙基；环烷基的实例包括但不限于环丙基、环丁基、环戊基、环己基、环庚基和环辛基。
本发明所述的“卤代烷基”包括单卤代烷基和多卤代烷基(其中所有卤原子可以相同或不同)。部分卤代的烷基是本文含义内的“卤代烷基”。 卤代烷基的实例包括三氟甲基、1,1-二氯乙基、1,2-二氯乙基、1,3-二溴-3,3-二氟丙基等。
本发明所述的“烷氧基”，是指上述烷基与氧原子相连所形成的基团，其中，氧原子具有自由成键能力，如甲氧基、乙氧基、丙氧基、丁氧基、戊氧基、异丙氧基、特丁氧基、环丙氧基、环己基氧基等。
本发明所述的“药学上可接受的盐”或“可药用盐”是指药学上可接受的酸和碱加成盐和溶剂化物。这类药学上可接受的盐包括与酸形成的盐，所述酸包括盐酸、磷酸、氢溴酸、硫酸、亚磺酸、甲酸、对甲苯磺酸、甲磺酸、硝酸、苯甲酸、柠檬酸、酒石酸、马来酸、氢碘酸、链状羧酸如乙酸、HOOC-(CH2)n-COOH(n＝0-4)等；也包括与碱形成的盐，这些盐的阳离子包括钠、钾、钙、铵离子等。
本发明中，“取代的”指本文中所定义的有机基团(其中包含有与氢原子键合的一个或多个键)被一个或多个与非氢原子或原子团键合的键取代，所述非氢原子或原子团即为取代基。
本发明化合物的取代基包括例如卤素、烷基(优选C1-8烷基、C1-6烷基或C1-4烷基)、烷氧基(优选C1-8烷氧基、C1-6烷氧基或C1-4烷氧基)、卤代烷基(优选C1-8卤代烷基、C1-6卤代烷基或C1-4卤代烷基)、卤代烷氧基(优选C1-8卤代烷氧基、C1-6卤代烷氧基或C1-4卤代烷氧基)、羟基、羟基烷基(优选羟基C1-8烷基、羟基C1-6烷基或羟基C1-4烷基)。
另一方面，本发明还涉及本文所描述化合物的荧光标记、自旋标记、重金属或同位素标记衍生物，它们不仅可用于成像，还可用于体内和体外的检测，定位和定量组织样品(包括人类)中的IDO酶，通过标记化合物的结合抑制，识别IDO酶配合基。因此，本发明进一步提供了含有这种标记化合物的IDO酶检测试剂或试剂盒。
本发明进一步提供本发明化合物的同位素标记化合物。所述本发明化合物的“同位素标记化合物”或“同位素标记”的本发明化合物是指本文所描述的化合物，其中一个或多个原子被同位素原子替代，该同位素原子的原子质量或质量数不同于通常在自然界中发现的(即自然发生 的)原子质量或质量数。适用的放射性核素可能包括但不限于2H(氘，也写成D)、3H(氚。也写成T)、11C、13C、14C、13N、15N、15O、17O、18O、18F、35S、36Cl、82Br、75Br、76Br、77Br、123I、124I、125I和131I。同位素标记的化合物中所含有的放射性同位素种类将取决于该同位素标记的化合物的具体应用。例如，对于体外IDO酶的标记和竞争试验，包含3H、14C、82Br、125I、131I、35S的化合物通常最有用。对于同位素成像应用，11C、18F、125I、123I、124I、131I、75Br、76Br或77Br一般最有用。
现有技术中已知的用放射性同位素标记有机化合物的方法同样适用于本发明的化合物。
在另一方面，本发明提供了制备本发明通式I化合物的方法：
Figure PCTCN2017116914-appb-000003
所述方法包括使式C化合物
Figure PCTCN2017116914-appb-000004
在有机溶剂和还原剂作用下反应形成通式I化合物，其中
R1、R2、R3、X和A如前面所定义。
本发明还提供了制备式C化合物的方法：
Figure PCTCN2017116914-appb-000005
包括使式A化合物
Figure PCTCN2017116914-appb-000006
与式B化合物
Figure PCTCN2017116914-appb-000007
在有机溶剂和偶联试剂的作用下偶联形成式C化合物，其中R1、R3、X和A如前面所定义。
特别地，本发明还提供了所述式C化合物作为用于制备本发明所述通式I化合物的中间体的用途。
式A化合物的制备
此外，本发明进一步提供了制备上述式A化合物的方法，包括以下步骤(a)-(d)：
(a)在钯催化作用下，化合物1与化合物2进行铃木交叉偶联得到 化合物3；
Figure PCTCN2017116914-appb-000008
(b)在碱的作用下，化合物3与化合物4进行Claisen-Schimidt缩合得到化合物5；
Figure PCTCN2017116914-appb-000009
(c)化合物5在醋酸条件下脱去三苯甲基并关环得到化合物6；
Figure PCTCN2017116914-appb-000010
(d)将化合物6在酸性条件下脱Boc保护得到中间体A：
Figure PCTCN2017116914-appb-000011
上述铃木交叉偶联反应(a)以钯为催化剂，如Pd(PPh3)4、Pd(dffp)Cl2、Pd(OAc)2、Pd(dba)3/PCy3、PdCl2、Pd(PPh3)4Cl2等。或者以Ni为催化剂，如NiCl2(dffp)、NiCl2(dffp)/Zn、NiCl2(dffp)/BuLi、NiCl2(PPh3)2/PPh3、NiCl2(NEt3)2、NiCl2(NEt3)2、NiCl(bipy)、Ni(TPPS)3、Ni(COD)2、NiCl2(PPh3)2/n-BuLi、Ni{P(OMe)3}2Cl2、NiCl2(PCy3)2等。以KOAc、K3PO4、K2CO3、NaOH、Ba(OH)2、Na2CO3、CsF或NaHCO3等为碱，反应温度为25到140℃，反应时间为4～72小时。反应所用溶剂为常用溶剂，如乙醇、THF、异丙醇、DMSO、DMF、二氧六环、甲苯、水和DME等。
上述Claisen-Schimidt缩合反应(b)可以以NaOH、NaOCH3、NaOEt等碱为催化剂，或者以酸为催化剂，如硫酸、盐酸、醋酸等。反应温度为0～80℃，反应时间为1～24小时。反应所用溶剂可以为常用溶剂，如THF、乙醇、水等。
上述反应(c)以醋酸、盐酸、三氟醋酸为脱保护和关环试剂，反应温度为0～90℃，反应时间为2～72小时。反应所用溶剂为常用溶剂，如水、甲醇、乙醇、异丙醇、四氢呋喃、二氯甲烷、丙酮等。
上述脱保护反应(d)以盐酸、三氟醋酸、对甲苯磺酸、硫酸、甲磺酸、三氟化硼乙醚等为脱保护试剂，反应温度为25～100℃，反应时间为20分钟～24小时。反应所用溶剂为常用溶剂，如水、甲醇、乙醇、乙酸乙酯、二氯甲烷、四氢呋喃、1,4-二氧六环、N,N-二甲基甲酰胺等。
式B化合物的制备
根据式B化合物结构中X的不同，分为两种情况：X为O；X为NR4。
当X为O时，式B化合物为化合物B(1)：
Figure PCTCN2017116914-appb-000012
当X为NR4时，式B化合物为化合物B(2)：
Figure PCTCN2017116914-appb-000013
式B(1)化合物可由市场直接购得或由如下合成方法制备：
Figure PCTCN2017116914-appb-000014
市场可购得的酚7与市场可购得的化合物8在碱性条件下发生亲核取代反应得到化合物9。该亲核取代反应是以NaOH、Na2CO3、K2CO3等碱为去酸剂，反应温度为25～140℃，反应时间为2～72小时。反应所用溶剂为常用溶剂，如水、甲醇、乙醇、乙腈、苯、二甲苯、丙酮、N,N'-二甲基甲酰胺、DMSO等。
所得化合物9在NaOH、LiOH等碱作用下脱乙酯得化合物B(1)。反应温度为0～60℃，反应时间为0.5～2小时。反应所用常用溶剂为水、甲醇、乙醇、四氢呋喃、N,N'-二甲基甲酰胺等。
式B(2)化合物可由市场直接购得或由如下合成方法制备：
Figure PCTCN2017116914-appb-000015
市场可购得的胺10与市场可购得的化合物8在碱性条件下发生亲核取代反应得到化合物11。该亲核取代反应是以NaOH、Na2CO3、K2CO3 等碱为去酸剂，反应温度为25～140℃，反应时间为2～72小时。反应所用溶剂为常用溶剂，如水、甲醇、乙醇、乙腈、苯、四氢呋喃、二甲苯、丙酮、N,N'-二甲基甲酰胺、DMSO等。
所得化合物11在NaOH、LiOH等碱作用下脱乙酯得式B(2)化合物。反应温度为0～60℃，反应时间为0.5～2小时。反应所用常用溶剂为水、甲醇、乙醇、四氢呋喃、N,N'-二甲基甲酰胺等。
在一个优选的实施方案中，本发明所述通式I化合物可由如下合成方法制备：
Figure PCTCN2017116914-appb-000016
式A化合物与式B化合物经缩合获得式C化合物。该缩合反应以肽缩合剂为催化剂，如1-乙基-3-(3-二甲胺丙基)碳二亚胺(EDC)、N,N'-二环己基碳二亚胺(DCC)、N,N'-羰基二咪唑(CDI)、O-(7-氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU)等。反应温度为0～60℃，反应时间为2～72小时。反应所用溶剂为常用溶剂，如苯、甲苯、四氢呋喃、二氧六环、二氯甲烷、氯仿、N,N'-二甲基甲酰胺等。必要时，也可以加入碱，如氢氧化钠、三乙胺或吡啶等。
式C中间体经还原剂如NaBH4、KBH4、NaBH(OAc)3、KBH(OAc)3、NaBH3CN等还原得到化合物I(R2＝-OH)。反应温度为0～60℃，反应时 间为2～72小时。反应所用溶剂为常用溶剂，如甲醇、乙醇、四氢呋喃等。
通式I(R2＝-OH)所示化合物通过Mitsunobu反应转化为通式I(R2＝-NH2)所示化合物。以偶氮二甲酸二乙酯(DEAD)和三苯基膦为活化试剂，邻苯二甲酰亚胺作为亲核试剂，然后经过肼解，得到通式I(R2＝-NH2)所示化合物。反应温度为0～25℃，反应时间为0.5～16小时。反应所用溶剂为常用溶剂，如四氢呋喃、乙醚、二氯甲烷、甲苯、乙酸乙酯、乙腈、N,N'-二甲基甲酰胺等。
通式(I)所述的化合物可以采用常见的分离方法进行纯化，如萃取、重结晶、柱层析等。
本发明所述的代表性化合物如下表1所示，其中化合物编号与实施例部分中的“实施例编号”相一致。
表1本发明的代表性化合物
Figure PCTCN2017116914-appb-000017
Figure PCTCN2017116914-appb-000018
Figure PCTCN2017116914-appb-000019
Figure PCTCN2017116914-appb-000020
Figure PCTCN2017116914-appb-000021
Figure PCTCN2017116914-appb-000022
具体实施方式
下面结合实例进一步阐明本发明的内容，但本发明的保护范围并不仅仅局限于这些实例。本发明所述的百分比除特别注明外，均为重量百分比。说明书中所描述的数值范围，如计量单位、反应条件、化合物物理状态或百分比，均是为了提供明白无误的书面参考。本领域熟练技术人员在实践本专利时，使用在此范围之外或有别于单个数值的温度、浓度、数量、碳原子数等，仍然可以得到预期的结果。
实施例中所使用的试剂缩写所代表的完整名称如下：
HATU      2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯
DIPEA     二异丙基乙基胺
DABCO     1,4-二氮杂二环[2.2.2]辛烷
HOBt      1-羟基苯并三唑
EDCI.HCl  1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐
DMF       N,N-二甲基甲酰胺
THF       四氢呋喃
EA        乙酸乙酯
DCM       二氯甲烷
实施例1
4-碘-1-三苯甲基-1H-咪唑的制备
Figure PCTCN2017116914-appb-000023
将4-碘-1H-咪唑(20g,103mmoles)溶于DMF(100mL)中，然后加入三乙胺(15.1mL,108mmoles)和三苯基甲基氯(27.8g,100mmoles)。反应液在室温下搅拌反应48h,然后倾倒到冰水中(500mL)，析出大量固体，过滤，烘干，得到白色固体4-碘-1-三苯甲基-1H-咪唑(40g,91.7mmol,88.9％)。
LC-MS(m/z):437(M+1)。
实施例2
2-(1-三苯甲基-1H-咪唑-4-基)苯甲醛的制备
Figure PCTCN2017116914-appb-000024
将4-碘-1-三苯甲基-1H-咪唑(38.7g,88.8mmol),(2-甲酰苯基)硼酸(20.0g,133mmol)和K3PO4(56.4g,266mmol)溶解到1,4-二氧六环(300mL)和水(60mL)中，反应液中鼓入氮气5min,然后加入Pd(PPh3)4(5.12g,4.44mmol)，并继续对反应液鼓入氮气5min。该反应在氮气保护下，升温90℃反应16h。反应结束后降温，垫硅藻土过滤，滤液用水(100mL)和EA(300mL)稀释，静置分层，水相用EA(300mL×2萃取；合并有机相，用水(100mL)和饱和食盐水(100mL×3)洗涤。有机相减压浓缩得到残留物，用PE/EA＝3/1过柱纯化，得到淡棕色粘稠油状物2-(1-三苯甲基-1H-咪唑-4-基)苯甲醛(15.0g,40.8％)。LC-MS(m/z):415(M+1)。
实施例3
(E)-4-(3-(2-(1-三苯甲基-1H-咪唑-4-基)苯基)丙烯酰基)哌啶-1-甲酸叔丁酯的制备
Figure PCTCN2017116914-appb-000025
将2-(1-三苯甲基-1H-咪唑-4-基)苯甲醛(4.0g,9.66mmol)溶解于无水THF(20mL)和无水EtOH(20mL)的混合溶液中，降温到0℃，然后加入EtONa(986mg,14.5mmol),4-乙酰基哌啶-1-甲酸叔丁酯(2.29g,10.1mmol)。然后反应液置于室温搅拌过夜，LCMS检测反应完全后， 用冰水(30mL)稀释，减压蒸出THF，抽滤，滤饼尽量抽干，得到白色固体粗品(E)-4-(3-(2-(1-三苯甲基-1H-咪唑-4-基)苯基)丙烯酰基)哌啶-1-甲酸叔丁酯(6.0g),直接用于下一步。
LC-MS(m/z):624(M+1)。
实施例4
4-(2-(5H-咪唑并[5,1-a])异吲哚-5-基)乙酰基)哌啶-1-甲酸叔丁酯的制备
Figure PCTCN2017116914-appb-000026
将上步所得粗品(E)-4-(3-(2-(1-三苯甲基-1H-咪唑-4-基)苯基)丙烯酰基)哌啶-1-甲酸叔丁酯(6.0g,粗品)溶于MeOH(80mL)中，加入冰乙酸(15mL)，然后升温到90℃搅拌过夜.反应结束后，降温到室温，用饱和碳酸钾溶液(30mL)调节pH到10。反应液用乙酸乙酯(100mL×3)萃取.合并有机相,用水(50mL)洗涤，饱和食盐水(50mL×3)洗涤。有机相用无水硫酸钠干燥，过滤，减压回收溶剂，残留物过柱(淋洗剂：CH2Cl2/MeOH＝50/1)纯化，得到淡黄色固体4-(2-(5H-咪唑并[5,1-a])异吲哚-5-基)乙酰基)哌啶-1-甲酸叔丁酯(2.7g,73.4％,两步总收率)。
LC-MS(m/z):382(M+1)。
实施例5
2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐的制备
Figure PCTCN2017116914-appb-000027
将4-(2-(5H-咪唑并[5,1-a])异吲哚-5-基)乙酰基)哌啶-1-甲酸叔丁酯(1.2g,3.15mmol)溶于1,4-二氧六环(20mL)，然后加入4N HCl/1,4-二氧 六环(4mL)。反应在室温下反应过夜，减压浓缩得到淡黄色固体粗品2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(1.2g)，直接用于下一步反应。
LC-MS(m/z):282(M+1)。
实施例6
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-苯氧乙基-1-酮的制备
Figure PCTCN2017116914-appb-000028
将2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(360mg，1.14mmol)，DIPEA(0.4mL，2.28mmol)溶于DCM(15mL)，冰浴搅拌，分批加入2-苯氧乙酸酐(340mg，1.18mmol)，慢慢升至室温反应6小时。反应液中加入饱和碳酸氢钠调PH至8-10，乙酸乙酯(50mL)萃取，用饱和食盐水(20mL)洗涤，20ml水洗涤.有机相用无水Na2SO4干燥，过滤，减压浓缩，残留物用5mL甲醇溶解，滴入50mL水中，析出固体，过滤获得化合物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-苯氧乙基-1-酮(250mg，收率52.9％)。
LC-MS(m/z):416(M+1)。
实施例7
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-苯氧乙基-1-酮的制备
Figure PCTCN2017116914-appb-000029
将1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-苯氧乙基-1-酮(250mg，0.60mmol)溶于20mL甲醇中，降温到0℃，分批加入NaBH4(68mg,1.8mmol)，室温反应过夜，加水淬灭，倒入水中，析出固体，过滤，所得固体溶于5mL甲醇中，倒入碳酸氢钠水溶液(50mL)中，析出固体，过滤获得1705(87mg，收率34.7％)。
1H NMR(400MHz,DMSO-d6):δ7.95(s,1H),7.60(d,J＝7.5Hz,1H),7.57(d,J＝7.5Hz,1H),7.38(t,J＝7.5Hz,1H),7.29-7.25(m,3H),7.12(s,1H),6.94-6.89(m,3H),5.41(t,J＝6.2Hz,1H),5.02(d,J＝5.2Hz,1H),4.81-4.73(m,2H),4.37(t,J＝13.0Hz,1H),3.87(t,J＝13.0Hz,1H),3.71-3.66(m,1H),2.96(t,J＝13.0Hz,1H),2.55(t,J＝13.0Hz,1H),2.10-2.02(m,1H),1.90-1.85(m,1H),1.80-1.77(m,1H),1.61-1.51(m,2H),1.34-1.10(m,2H).
LC-MS(m/z):418(M+1)。
实施例8
2-((四氢呋喃-3-基)氧基)乙酸乙酯的制备
Figure PCTCN2017116914-appb-000030
在0℃，NaH(6.81g,170mmol)加入到THF(60mL),然后加入3-羟基四氢呋喃(5.00g,56.8mmol)后，在0℃搅拌反应0.5h,加入2-溴乙酸乙酯(19.0g,114mmol).反应液在室温下搅拌过夜，浓缩，残留物用EA(100mL)稀释后，用水(50mL)洗涤，饱和食盐水(50mL)洗涤.有机 相用无水硫酸钠干燥，过滤，减压浓缩，残留物用PE/EA＝20/1～10/1过柱纯化，得到黄色油状物2-((四氢呋喃-3-基)氧基)乙酸乙酯(3.50g,20.1mmol,35.4％)。
LC-MS(m/z):175(M+1)。
实施例9
2-((四氢呋喃-3-基)氧基)乙酸的制备
Figure PCTCN2017116914-appb-000031
2-((四氢呋喃-3-基)氧基)乙酸乙酯(1.20g,6.90mmol)和KOH(10.4mL,20.7mmol,2.0M in water)溶解到MeOH(30mL)中，在80℃搅拌反应6h.反应液浓缩掉一部分后，用浓盐酸调节pH＝5.0，混合物用DCM/MeOH(v/v,5/1,100mL)萃取。有机相用无水硫酸钠干燥，过滤，减压浓缩得到2-((四氢呋喃-3-基)氧基)乙酸(350mg),直接用于下一步反应。
LC-MS(m/z):147(M+1)。
实施例10
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((四氢呋喃-3-基)氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000032
2-((四氢呋喃-3-基)氧基)乙酸(50mg,0.158mmol),2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg，粗品),HATU (120mg,0.316mmol)和DIPEA(61.1mg,0.474mmol)溶于DMF(5mL)中，室温搅拌反应过夜，用乙酸乙酯(50mL)稀释，水(10mL)洗涤，饱和食盐水(10mL×3)洗涤。有机相用无水硫酸钠干燥，过滤，浓缩得到残留物，用DCM/MeOH＝10/1过柱纯化得到1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((四氢呋喃-3-基)氧基)乙基-1-酮(100mg),直接用于下一步反应。
LC-MS(m/z):410(M+1)。
实施例11
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-((四氢呋喃-3-基)氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000033
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((四氢呋喃-3-基)氧基)乙基-1-酮(100mg,crude)溶于甲醇(10mL)中，降温到0℃，缓慢加入硼氢化钠(18.0mg,0.474mmol)，在该温度下搅拌反应1h,用水(10mL)淬灭反应。反应液浓缩得到残留物，用制备色谱制得白色固体化合物1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-((四氢呋喃-3-基)氧基)乙基-1-酮(10.0mg,0.024mmols,2.47％的三步总收率)。
1H NMR(500MHz,DMSO-d6)δ7.94和7.91(s,1H),7.60(d,J＝7.5Hz,1H),7.56和7.50(d,J＝7.7Hz,1H),7.38(t,J＝7.5Hz,1H),7.27(t,J＝7.4Hz,1H),7.14和7.12(s,1H),5.48–5.34(m,1H),5.14和5.02(d,J＝4.9Hz,1H),4.45–4.29(m,1H),4.19–4.00(m,3H),3.88–3.75(m,1H),3.75–3.56(m,5H),2.88(t,J＝12.9Hz,1H),2.45(t,J＝12.2Hz,1H),2.26–2.02(m,1H),1.97–1.83(m,3H),1.82–1.71(m,1H),1.61–1.47(m,2H),1.28–1.19(m,1H),1.18–1.04(m,1H)ppm.
LC-MS(m/z):412(M+1)。
实施例12
3-甲氧基-9H-咔唑的制备
Figure PCTCN2017116914-appb-000034
6-甲氧基-2,3,4,9-四氢-1H-咔唑(3.00g,14.9mmol)和I2(948mg,3.73mmol)溶于DMSO(30mL)中，加热升温到90℃过夜反应.反应结束后，用乙酸乙酯(300mL)稀释，有机相用饱和食盐水(50mL×3)洗涤，无水硫酸钠干燥，过滤，减压浓缩，残留物用PE/EA＝10/1～5/1淋洗剂过柱纯化，得到白色固体3-甲氧基-9H-咔唑(345mg,1.75mmol,11.8％)。
LC-MS(m/z):198(M+1)。
实施例13
3-羟基-9H-咔唑的制备
Figure PCTCN2017116914-appb-000035
3-甲氧基-9H-咔唑(345mg,1.75mmol)溶于氢溴酸(5mL)和醋酸(20mL)配成的溶液中，加热回流反应2h，降温到室温，浓缩掉部分溶剂，用饱和碳酸氢钠溶液调节pH＝9.0，用乙酸乙酯(50mL)萃取。有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，减压浓缩得到3-羟基-9H-咔唑(300mg,1.64mmol,93.7％)。
LC-MS(m/z):184(M+1)。
实施例14
2-((9H-咔唑-3-基)氧基)乙酸乙酯的制备
Figure PCTCN2017116914-appb-000036
3-羟基-9H-咔唑(300mg,1.64mmol),2-溴乙酸乙酯(329mg,1.97mmol)和K2CO3(272mg,1.97mmol)溶于DMF(10mL)中，室温搅拌过夜。反应液用乙酸乙酯(80mL)稀释,用饱和食盐水(20mL×3)洗涤。有机相用无水硫酸钠干燥，过滤，减压浓缩，残留物用乙酸乙酯/石油醚＝1/10的淋洗剂过柱，得到2-((9H-咔唑-3-基)氧基)乙酸乙酯(340mg,1.26mmol，77％),直接用于下一步反应。
LC-MS(m/z):270(M+1)。
实施例15
2-((9H-咔唑-3-基)氧基)乙酸的制备
Figure PCTCN2017116914-appb-000037
2-((9H-咔唑-3-基)氧基)乙酸乙酯(154mg,0.572mmol)溶于2N氢氧化钠溶液(0.86mL,1.72mmol)，加入MeOH(5mL)，室温搅拌反应4h，反应液浓缩掉部分溶剂，用浓盐酸调节pH＝5.0，用乙酸乙酯(30mL)萃取。有机相用食盐水(15mL)洗涤,无水硫酸钠干燥，过滤，减压浓缩，得到2-((9H-咔唑-3-基)氧基)乙酸粗品(200mg),直接用于下一步反应。
LC-MS(m/z):242(M+1)。
实施例16
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((9H-咔唑-3- 基)氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000038
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((9H-咔唑-3-基)氧基)乙基-1-酮(90mg,粗品)是由2-((9H-咔唑-3-基)氧基)乙酸(45.7mg,0.190mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例10中的类似步骤制备而成。
LC-MS(m/z):505(M+1)。
实施例17
2-((9H-咔唑-3-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000039
白色固体2-((9H-咔唑-3-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(21mg,两步收率26.3％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((9H-咔唑-3-基)氧基)乙基-1-酮(90mg,粗品)按照实施例11中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ11.05(s,1H),8.04(d,J＝7.6Hz,1H),7.95和7.92(s,1H),7.66(s,1H),7.63–7.47(m,2H),7.44(d,J＝8.1Hz,1H),7.42–7.31(m,3H),7.27(t,J＝7.5Hz,1H),7.18–7.07(m,2H),7.04(d,J＝8.8Hz,1H),5.48–5.34(m,1H),5.19–5.01(m,1H),4.90– 4.73(m,2H),4.48–4.34(m,1H),4.06–3.90(m,1H),3.76–3.66(m,1H),3.01(t,J＝13.2Hz,1H),2.54(t,J＝12.7Hz,1H),2.27–2.08(m,1H),1.96–1.50(m,4H),1.43–1.31(m,1H),1.25–1.10(m,1H)ppm.
LC-MS(m/z):507(M+1)。
实施例18
2-(哒嗪-4-基氧基)乙酸乙酯的制备
Figure PCTCN2017116914-appb-000040
将4-羟基哒嗪(2.00g,20.8mmol),2-溴乙酸乙酯(5.21g,31.2mmol)和K2CO3(4.31g,31.2mmol)加入到乙腈(30mL)中，加热回流反应4h。反应液减压浓缩，用乙酸乙酯/石油醚＝1/10过柱纯化，得到2-(哒嗪-4-基氧基)乙酸乙酯(1.40g,7.69mmol,37.0％)。
LC-MS(m/z):183(M+1)。
实施例19
2-(哒嗪-4-基氧基)乙酸的制备
Figure PCTCN2017116914-appb-000041
2-(哒嗪-4-基氧基)乙酸(300mg,粗品)是由2-(哒嗪-4-基氧基)乙酸乙酯(200mg,1.10mmol)按照实施例15中的类似步骤制备而成。
LC-MS(m/z):155(M+1)。
实施例20
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(哒嗪-4-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000042
将2-(哒嗪-4-基氧基)乙酸(50mg,0.158mmol),2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,粗品),HATU(120mg,0.316mmol)和DIPEA(61.1mg,0.474mmol)加入到5mL DMF中，室温搅拌过夜。反应液用乙酸乙酯50ml稀释，接着用10ml水洗涤，饱和食盐水(10mL×3)洗涤。有机相用无水Na2SO4干燥，过滤，减压浓缩，残留物用二氯甲烷/甲醇＝10/1过柱纯化，得到化合物粗品1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(哒嗪-4-基氧基)乙基-1-酮(75mg,粗品),直接用于下一步反应。
LC-MS(m/z):418(M+1)。
实施例21
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(哒嗪-4-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000043
将实施例20所得的1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(哒嗪-4-基氧基)乙基-1-酮(75mg,crude)溶于10mL甲醇，降温到0℃，加入NaBH4(18.0mg,0.474mmol)，在该温度下反应1h，用水(10mL)淬灭反应，减压浓缩得到残留物，用制备色谱制得白色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(哒嗪-4- 基氧基)乙基-1-酮(28.0mg,0.067mmols,两步总收率20.6％)。
1H NMR(500MHz,DMSO-d6)δ8.10(dd,J＝7.8,1.5Hz,1H),7.98–7.89(m,1H),7.73(d,J＝3.1Hz,1H),7.61(d,J＝7.6Hz,1H),7.57和7.51(d,J＝7.9Hz,1H),7.39(t,J＝7.5Hz,1H),7.28(t,J＝7.5Hz,1H),7.15和7.12(s,1H),6.35(dd,J＝7.8,3.1Hz,1H),5.48–5.36(m,1H),5.21–5.01(m,3H),4.42–4.27(m,1H),3.88–3.75(m,1H),3.74–3.67(m,1H),2.98(t,J＝13.1Hz,1H),2.55(t,J＝12.6Hz,1H),2.27–2.03(m,1H),1.92–1.52(m,4H),1.38–1.25(m,1H),1.20–1.10(m,1H)ppm。
LC-MS(m/z):420(M+1)。
实施例22
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(喹啉-6-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000044
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(喹啉-6-基氧基)乙基-1-酮(50mg,粗品)是由2-(喹啉-6-基氧基)乙酸(54mg,0.267mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例20中的类似步骤制备而成。
LC-MS(m/z):467(M+1)。
实施例23
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(喹啉-6-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000045
白色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(喹啉-6-基氧基)乙基-1-酮(30mg,两步收率58％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(喹啉-6-基氧基)乙基-1-酮(50mg,粗品)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ9.32和9.30(s,1H),8.85(d,J＝3.3Hz,1H),8.39(d,J＝7.9Hz,1H),8.00(d,J＝9.2Hz,1H),7.94–7.92(m,1H),7.87(d,J＝7.2Hz,1H),7.74和7.67(d,J＝8.0Hz,1H),7.61(dd,J＝8.3,4.5Hz,1H),7.58–7.52(m,3H),7.41(d,J＝2.5Hz,1H),5.79(t,J＝8.0Hz,1H),5.06–4.89(m,2H),4.40(t,J＝13.0Hz,1H),4.01–3.86(m,3H),3.03(t,J＝12.8Hz,1H),2.65–2.53(m,1H),2.23–2.12(m,1H),2.12–1.99(m,1H),1.81(t,J＝15.3Hz,1H),1.69–1.53(m,2H),1.43–1.24(m,1H),1.22–1.06(m,1H)ppm.
LC-MS(m/z):469(M+1)。
实施例24
2-((1H-吲哚-5-基)氧基)乙酸的制备
Figure PCTCN2017116914-appb-000046
将2-((1H-吲哚-5-基)氧基)乙酸乙酯(200mg,0.913mmol)溶于甲醇(10mL)中，降温到0℃，然后加入2.0M氢氧化钠溶液(0.915mL,1.83mmol)。反应液在室温下反应2h，加入20ml水，冻干，得到白色固体2-((1H-吲哚-5-基)氧基)乙酸(200mg,粗品),直接用于下一步反应。
LC-MS(m/z):192(M+1)。
实施例25
2-((1H-吲哚-5-基)氧基)-1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-乙基-1-酮的制备
Figure PCTCN2017116914-appb-000047
2-((1H-吲哚-5-基)氧基)-1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-乙基-1-酮(70mg,crude)是由2-((1H-吲哚-5-基)氧基)乙酸(90.4mg,0.473mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例20中的类似步骤制备而成。
LC-MS(m/z):455(M+1)。
实施例26
2-((1H-吲哚-5-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-乙基-1-酮的制备
Figure PCTCN2017116914-appb-000048
白色固体2-((1H-吲哚-5-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-乙基-1-酮(22mg,两步收率27％)是由2-((1H-吲哚-5-基)氧基)-1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-乙基-1-酮(70mg,粗品)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ10.91(s,1H),7.94和7.92(s,1H),7.66–7.45(m,2H),7.38(t,J＝7.5Hz,1H),7.28–7.25(m,3H),7.14和7.12(s,1H),7.03(s,1H),6.74(d,J＝8.5Hz,1H),6.31(s,1H),5.38(t,J＝6.0Hz,1H),5.12–5.00(m,1H),4.77–4.57(m,2H),4.47–4.28(m,1H),4.01–3.85(m,1H),3.72-3.68(m,1H),2.97(t,J＝12.3Hz,1H),2.26–1.98(m,1H),1.94–1.72(m,2H),1.67–1.48(m,2H),1.34–1.08(m,2H)ppm.
LC-MS(m/z)457(M+1)。
实施例27
3-(2-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-氧乙氧基)吡咯烷-1-甲酸叔丁酯的制备
Figure PCTCN2017116914-appb-000049
3-(2-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-氧乙氧基)吡咯烷-1-甲酸叔丁酯(60mg,53.6％)是由2-((1-(叔丁氧基羰基)吡咯烷-3-基)氧基)乙酸(59.5mg,0.243mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(70mg,0.221mmol)按照实施例10中的类似步骤制备而成。
LC-MS(m/z):509(M+1)。
实施例28
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(吡咯烷-3-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000050
3-(2-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-氧乙氧基)吡咯烷-1-甲酸叔丁酯(60mg,0.118mmol)溶于1，4-二氧六环(10mL)中，降温到0℃，加入1，4-二氧六环的盐酸溶液(0.118mL,4M,0.472mmol)。反应液在室温搅拌反应2h，用饱和碳酸氢钠调节pH＝7至8。反应液用乙酸乙酯(30mL×3)萃取，合并有机相，饱和食盐水洗，无水硫酸钠干燥，过滤，减压浓缩得到淡黄色固体产物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(吡咯烷-3-基氧基)乙基-1-酮粗品(60mg),直接用于下一步反应。
LC-MS(m/z):409(M+1)。
实施例29
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(吡咯烷-3-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000051
白色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(吡咯烷-3-基氧基)乙基-1-酮(19mg,两步收率39.2％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(吡咯烷-3-基氧基)乙基-1-酮(60mg,crude)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ7.94和7.90(s,1H),7.60(d,J＝7.5Hz,1H),7.57和7.49(d,J＝7.5Hz,1H),7.38(t,J＝7.5Hz,1H),7.27(t,J＝7.6Hz,1H),7.14和7.12(s,1H),5.43和5.39(d,J＝7.0Hz,1H), 5.02(brs,1H),4.43–4.23(m,1H),4.17–3.94(m,3H),3.90–3.74(m,1H),3.72–3.63(m,1H),3.30–3.12(m,3H),2.94–2.59(m,3H),2.48–2.34(m,1H),2.08-2.07(m,1H),1.97–1.62(m,4H),1.58-1.53(m,2H),1.24–1.10(m,2H).
LC-MS(m/z):411(M+1)。
实施例30
2-((9H-嘌呤-6-基)氧基)乙酸乙酯的制备
Figure PCTCN2017116914-appb-000052
6-氯-9H-嘌呤(3.08g,20.0mmol)和DABCO(6.72g,60.0mmol)溶于DMSO(30mL)中，在20℃搅拌反应4h，然后升温到35℃反应0.5h.该混合物加入到2-羟基乙酸乙酯(20.8g,200mmol)，NaH(6.40g,160mmol)和DMSO(30mL)生成的反应液中。该反应液在室温搅拌过夜，浓缩，残留物用2-甲基四氢呋喃(1L)稀释，用饱和食盐水(100mL×3)洗涤。有机相用无水硫酸钠干燥，过滤，浓缩，残留物用EA/PE＝1：1过柱纯化得到黄色固体化合物2-((9H-嘌呤-6-基)氧基)乙酸乙酯(100mg,0.450mmol,2.25％)。
LC-MS(m/z):223(M+1)。
实施例31
2-((9H-嘌呤-6-基)氧基)乙酸的制备
Figure PCTCN2017116914-appb-000053
将2-((9H-嘌呤-6-基)氧基)乙酸乙酯(100mg,0.450mmol)、2N氢氧 化钠溶液(0.67mL,1.35mmol)加入到甲醇(5mL)中，室温搅拌过夜，浓缩，残留物用浓盐酸调节pH＝5.0，浓缩得到2-((9H-嘌呤-6-基)氧基)乙酸粗品(200mg)，直接用于下一步反应。
LC-MS(m/z):195(M+1)。
实施例32
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((7H-嘌呤-6-基)氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000054
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((7H-嘌呤-6-基)氧基)乙基-1-酮(90mg,粗品)是由2-((9H-嘌呤-6-基)氧基)乙酸(36.7mg,0.189mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例10中的类似步骤制备而成。
LC-MS(m/z):458(M+1)。
实施例33
2-((7H-嘌呤-6-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000055
白色固体2-((7H-嘌呤-6-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(21mg,两步收率29％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((7H-嘌呤-6-基)氧基)乙基-1-酮(90mg,粗品)按照实施例11中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ13.44(s,1H),8.42–8.37(m,2H),8.01–7.91(m,1H),7.68–7.47(m,2H),7.39(t,J＝7.5Hz,1H),7.28(t,J＝7.6Hz,1H),7.15和7.12(s,1H),5.49–5.37(m,1H),5.37–5.23(m,2H),5.17和5.06(t,J＝6.0Hz,1H),4.38–4.24(m,1H),3.94–3.80(m,1H),3.78–3.66(m,1H),3.01(t,J＝12.9Hz,1H),2.25–2.05(m,1H),1.95–1.50(m,4H),1.44–1.31(m,1H),1.21–1.08(m,1H)ppm.
LC-MS(m/z):460(M+1)。
实施例34
2-(苯并[d]恶唑啉-2-基氧基)乙酸乙酯的制备
Figure PCTCN2017116914-appb-000056
在0℃下，NaH(1.00g,25.0mmol)加入到THF(30mL)中，然后加入2-羟基乙酸乙酯(2.08g,20.0mmol).反应液在0℃搅拌0.5h,然后加入2-氯苯并[d]恶唑啉(1.53g,10.0mmol).反应液加热回流反应3h，减压回收部分溶剂后，用乙酸乙酯EA(100mL)稀释。乙酸乙酯相用水(50mL)洗涤，饱和食盐水(50mL)洗涤。有机相用无水硫酸钠干燥，过滤，减压浓缩，残留物用乙酸乙酯/石油醚＝1/20—1/10的淋洗剂过柱，得到油状化合物2-(苯并[d]恶唑啉-2-基氧基)乙酸乙酯(1.11g,5.02mmols,50.2％)。
LC-MS(m/z):222(M+1)。
实施例35
2-(苯并[d]恶唑啉-2-基氧基)乙酸的制备
Figure PCTCN2017116914-appb-000057
取部分2-(苯并[d]恶唑啉-2-基氧基)乙酸乙酯(100mg,0.452mmol)溶于氢氧化钠溶液(0.23mL,0.452mmol,2.0M)中，加入甲醇(5mL)，室温搅拌反应过夜。反应液冷冻干燥得到2-(苯并[d]恶唑啉-2-基氧基)乙酸(200mg，crude)粗品，直接用于下一步反应。
LC-MS(m/z):194(M+1)。
实施例36
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(苯并[d]恶唑啉-2-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000058
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(苯并[d]恶唑啉-2-基氧基)乙基-1-酮(90mg,crude)是由2-(苯并[d]恶唑啉-2-基氧基)乙酸(36.7mg,0.190mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例10中的类似步骤制备而成。
LC-MS(m/z):457(M+1)。
实施例37
2-(苯并[d]恶唑啉-2-基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000059
白色固体2-(苯并[d]恶唑啉-2-基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(14mg,两步收率19.3％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(苯并[d]恶唑啉-2-基氧基)乙基-1-酮(90mg,粗品)按照实施例11中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ7.96和7.93(s,1H),7.62–7.50(m,3H),7.45(d,J＝7.5Hz,1H),7.39(t,J＝7.5Hz,1H),7.32–7.19(m,3H),7.15和7.13(s,1H),5.48–5.37(m,1H),5.36–5.24(m,2H),5.16和5.05(t,J＝6.9Hz,1H),4.37–4.27(m,1H),3.81–3.66(m,2H),2.99(t,J＝12.5Hz,1H),2.54(t,J＝12.5Hz,1H),2.26–1.74(m,3H),1.68–1.50(m,2H),1.42–1.28(m,1H),1.19–1.11(m,1H)ppm.
LC-MS(m/z):459(M+1)。
实施例38
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-氯-4-氟苯氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000060
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-氯-4-氟苯氧基)乙基-1-酮(90mg,粗品)是由2-(3-氯-4-氟苯氧基)乙酸(38.6mg,0.189mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例20中的类似步骤制备而成。
LC-MS(m/z):468(M+1)。
实施例39
2-(3-氯-4-氟苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000061
白色固体2-(3-氯-4-氟苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(10mg,两步收率13.5％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-氯-4-氟苯氧基)乙基-1-酮(90mg,粗品)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ7.94和7.91(s,1H),7.60(d,J＝7.6Hz,1H),7.56和7.50(d,J＝7.5Hz,1H),7.38(t,J＝7.4Hz,1H),7.35–7.23(m,2H),7.19–7.07(m,2H),6.94–6.89(m,1H),5.47–5.32(m,1H),5.15–5.01(m,1H),4.91–4.74(m,2H),4.36(t,J＝16.6Hz,1H),3.86–3.75(m,1H),3.74–3.64(m,1H),2.94(t,J＝12.4Hz,1H),2.26–2.02(m,1H),1.95–1.74(m,2H),1.65–1.48(m,2H),1.37–1.21(m,1H),1.20–1.08(m,1H)ppm.
LC-MS(m/z):470(M+1)。
实施例40
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-氯苯氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000062
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-氯苯氧基)乙基-1-酮(55mg,86％)是由2-(3-氯苯氧基)乙酸(32mg,0.17mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(40mg,0.142mmol)按照实施例20中的类似步骤制备而成。
LC-MS(m/z):450(M+1)。
实施例41
2-(3-氯苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000063
白色固体2-(3-氯苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(13mg,30％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-氯苯氧基)乙基-1-酮(50mg,0.11mmol)按照实施例21中的类似步骤制备而成。
1H NMR(400MHz,CDCl3)δ7.808(s,1H)7.543(d,J＝7.2Hz,1H),7.319-7.418(m,2H),7.238-7.275(m,1H),7.164-7.205(m,2H),6.921-6.959(m,2H),6.816(d,J＝8Hz,1H),5.360-5.503(m,1H),4.609-4.695(m,3H),3.701-3.804(m,1H),3.020(t,J＝12.8Hz,1H),2.558(t,J＝12.8Hz,1H),2.057-2.292(m,2H),1.845-1.883(m,1H),1.52-1.714(m,2H),1.26-1.338(m,2H).
LC-MS(m/z):452(M+1)。
实施例42
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(4-氟苯氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000064
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(4-氟苯氧基)乙基-1-酮(180mg,crude)是由2-(4-氟苯氧基)乙酸(40.8mg,0.24mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(63.4mg,0.2mmol)按照实施例20中的类似步骤制备而成。
LC-MS(m/z):434(M+1)。
实施例43
2-(4-氟苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000065
白色固体2-(4-氟苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(20mg,两步收率23％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(4-氟苯氧基)乙基-1-酮(100mg,粗品)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ7.94和7.91(s,1H),7.60(d,J＝7.5Hz,1H),7.56和7.50(d,J＝7.5Hz,1H),7.38(t,J＝7.5Hz,1H),7.27(t,J＝7.4Hz,1H),7.16–7.03(m,3H),6.92(dd,J＝8.3,4.2Hz,2H),5.47–5.33(m,1H),5.17–4.98(m,1H),4.85–4.67(m,2H),4.45–4.31(m,1H),3.92–3.77(m,1H),3.75–3.63(m,1H),2.95(t,J＝13.0Hz,1H),2.25–1.99(m,1H),1.93–1.83(m,1H),1.79(d,J＝12.9Hz,1H),1.64–1.48(m,2H),1.36–1.22(m,1H),1.20–1.05(m,1H)ppm.
LC-MS(m/z):436(M+1)。
实施例44
4-(1-羟基-2-(5H-咪唑并[5,1-a])异吲哚-5-基)乙基)哌啶-1-甲酸叔丁酯的制备
Figure PCTCN2017116914-appb-000066
4-(1-羟基-2-(5H-咪唑并[5,1-a])异吲哚-5-基)乙基)哌啶-1-甲酸叔丁酯是由4-(2-(5H-咪唑并[5,1-a])异吲哚-5-基)乙酰基)哌啶-1-甲酸叔丁酯按照实施例11中的类似步骤制备而成。
LC-MS(m/z):384(M+1)。
实施例45
2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)-乙基-1-醇盐酸盐的制备
Figure PCTCN2017116914-appb-000067
将4-(1-羟基-2-(5H-咪唑并[5,1-a])异吲哚-5-基)乙基)哌啶-1-甲酸叔丁酯(380mg,1mmol)溶于二氯甲烷(10mL)，然后加入4N HCl/1,4-二氧六环(5mL)。在0℃下反应4小时，减压浓缩得到黄色油状物粗品2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)-乙基-1-醇盐酸盐(241mg,85％),直接用于下一步反应。
LC-MS(m/z):284(M-HCl+1)。
实施例46
2-(3,4-二甲基苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000068
黄色固体2-(3,4-二甲基苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(40mg,35.7％)是由2-(3,4-二甲基苯氧基)乙酸(25mg,0.139mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)-乙基-1-醇盐酸盐(35.7mg,0.126mmol)按照实施例20中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ7.94和7.91(s,1H),7.60(d,J＝7.6Hz,1H),7.56和7.50(d,J＝7.5Hz,1H),7.38(t,J＝7.4Hz,1H),7.30–7.25(m,1H),7.14和7.12(s,1H),6.99(d,J＝8.2Hz,1H),6.71(s,1H),6.62(d,J＝8.2Hz,1H),5.46–5.35(m,1H),5.17–4.96(m,1H),4.75–4.61(m,2H),4.43–4.27(m,1H),3.95–3.79(m,1H),3.74–3.63(m,1H),3.00–2.90(m,1H),2.15(s,3H),2.11(s,3H),1.89–1.05(m,7H)ppm.
LC-MS(m/z):446(M+1)。
实施例47
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-三氟甲基苯氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000069
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-三氟甲基苯氧基)乙基-1-酮(90mg,粗品)是由2-(3-三氟甲基苯氧基)乙酸(41.6mg,0.189mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例20中的类似步骤制备而成。
LC-MS(m/z):484(M+1)。
实施例48
2-(3-三氟甲基苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000070
白色固体2-(3-三氟甲基苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(10mg,两步收率15％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(3-三氟甲基苯氧基)乙基-1-酮(90mg,crude)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ7.94和7.91(s,1H),7.60(d,J＝7.6Hz,1H),7.58–7.47(m,2H),7.38(t,J＝7.5Hz,1H),7.30–7.25(m,2H),7.21(d,J＝7.6Hz,2H),7.14和7.12(s,1H),5.47–5.34(m,1H), 5.17–5.00(m,1H),4.99–4.85(m,2H),4.42–4.31(m,1H),3.92–3.78(m,1H),3.75–3.64(m,1H),2.96(t,J＝12.8Hz,1H),2.24–2.02(m,1H),1.92–1.50(m,4H),1.36–1.24(m,1H),1.19–1.04(m,1H)ppm.LC-MS(m/z)486(M+1)。
实施例49
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(环己基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000071
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(环己基氧基)乙基-1-酮(80mg,粗品)是由2-(环己基氧基)乙酸(30mg,0.189mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例20中的类似步骤制备而成。
LC-MS(m/z):422(M+1)。
实施例50
2-(环己基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000072
白色固体2-(环己基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(10mg,两步收率15％)是由1-(4-(2-(5H- 咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(环己基氧基)乙基-1-酮(80mg,粗品)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ7.93和7.90(s,1H),7.60(d,J＝7.5Hz,1H),7.56和7.50(d,J＝7.5Hz,1H),7.38(t,J＝7.5Hz,1H),7.27(t,J＝7.5Hz,1H),7.14和7.11(s,1H),5.47–5.33(m,1H),5.11和5.00(t,J＝5.0Hz,1H),4.36(t,J＝13.9Hz,1H),4.14–4.00(m,2H),3.95–3.80(m,1H),3.67(t,J＝9.9Hz,1H),3.30–3.20(m,1H),2.88(t,J＝12.8Hz,1H),2.47–2.39(m,1H),2.25–2.01(m,1H),1.93–1.71(m,4H),1.63(s,2H),1.59–1.48(m,2H),1.48–1.39(m,1H),1.27–1.07(m,7H)ppm.
LC-MS(m/z):424(M+1)。
实施例51
1-(二苯并[b,d]呋喃-2-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000073
二苯并[b,d]呋喃(5.0g,29mmol)溶于氯仿(50mL)中，加入三氯化铝AlCl3(4.8g,35.6mmol)和乙酰氯(2.8g,35.6mmol)的氯仿(50mL)溶液中。反应液在室温下搅拌反应45min，倾倒到冰水(100mL)和1NHCl(50mL)配成的溶液中.水相用氯仿(2x 30mL)萃取,合并有机相，用无水硫酸钠干燥，真空浓缩，残留物用石油醚/乙酸乙酯＝20：1的淋洗剂过柱纯化，得到白色固体1-(二苯并[b,d]呋喃-2-基)乙基-1-酮(5.2g,90％)，直接用于下一步反应。
LC-MS(m/z):211(M+1)。
实施例52
乙酸二苯并[b,d]呋喃-2-酯的制备
Figure PCTCN2017116914-appb-000074
1-(二苯并[b,d]呋喃-2-基)乙基-1-酮(5.2g,24.8mmol)溶于二氯甲烷(150mL)中，降温到0℃，加入三氟乙酸(8.5g,75mmol)和mCPBA(6.25g,28mmol)。反应液室温下搅拌反应3d，用硫酸亚铁淬灭，水洗，有机相用无水硫酸钠干燥，减压浓缩得到乙酸二苯并[b,d]呋喃-2-酯粗品。
LC-MS(m/z):227(M+1)。
实施例53
2-羟基二苯并[b,d]呋喃的制备
Figure PCTCN2017116914-appb-000075
乙酸二苯并[b,d]呋喃-2-酯粗品溶于甲醇(100mL)中，加入甲醇钠(4g,75mmol)，然后室温搅拌反应20min。反应液用2N HCl(50mL)淬灭，减压回收有机溶剂，残留物用水稀释，用氯仿(2x 50mL)萃取,合并有机相，用无水硫酸钠干燥，过滤，减压浓缩，残留物用石油醚/乙酸乙酯＝10/1淋洗剂过柱纯化，得到灰黄色固体2-羟基二苯并[b,d]呋喃(3.6g,78％)。
LC-MS(m/z):185(M+1)。
实施例54
2-(二苯并[b,d]呋喃-2-基氧基)乙酸乙酯的制备
Figure PCTCN2017116914-appb-000076
2-羟基二苯并[b,d]呋喃(1.84g,10mmol),溴乙酸乙酯(2g,12mmol)和K2CO3(2.726g,20mmol)溶于乙腈(50mL)中，室温搅拌过夜，反应液过滤，滤液浓缩，残留物用乙酸乙酯/石油醚＝1/10过柱，得到2-(二苯 并[b,d]呋喃-2-基氧基)乙酸乙酯(2.5g,92％),直接用于下一步反应。
LC-MS(m/z):271(M+1)。
实施例55
2-(二苯并[b,d]呋喃-2-基氧基)乙酸的制备
Figure PCTCN2017116914-appb-000077
2-(二苯并[b,d]呋喃-2-基氧基)乙酸乙酯(2.5g,9.3mmol)溶于甲醇(30ml)和THF(30ml)的混合物溶剂中，加入氢氧化钠溶液(30ml,2N,60mmol),反应液室温搅拌过夜。停止反应，分出有机相，残留物中加入水(50ml)，用1N盐酸调节pH to 2～3，析出固体，过滤，滤饼用水(50ml)洗涤，干燥得到白色固体2-(二苯并[b,d]呋喃-2-基氧基)乙酸(2g,89％)，直接用于下步反应。
LC-MS(m/z):243(M+1)。
实施例56
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(二苯并[b,d]呋喃-2-基氧基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000078
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(二苯并[b,d]呋喃-2-基氧基)乙基-1-酮(90mg,crude)是由2-(二苯并[b,d]呋喃-2-基氧基)乙酸(46mg,0.189mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(50mg,0.158mmol)按照实施例20中的类似步骤 制备而成。
LC-MS(m/z):506(M+1)。
实施例57
2-(二苯并[b,d]呋喃-2-基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000079
2-(二苯并[b,d]呋喃-2-基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(36mg,两步收率45％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(二苯并[b,d]呋喃-2-基氧基)乙基-1-酮(90mg,粗品)按照实施例21中的类似步骤制备而成。
1H NMR(500MHz,DMSO-d6)δ8.09(d,J＝7.0Hz,1H),7.96和7.92(s,1H),7.69–7.66(m,2H),7.63–7.49(m,4H),7.38(t,J＝6.5Hz,2H),7.27(t,J＝7.5Hz,1H),7.14-7.10(m,2H),5.41–5.39(m,1H),5.17和5.06(t,J＝5.5Hz,1H),4.93-4.85(m,2H),4.41(t,J＝14.5Hz,1H),3.93(t,J＝16.3Hz,1H),3.72–3.71(m,1H),3.01(t,J＝12.8Hz,1H),2.55–2.52(m,1H),2.11–2.05(m,1H),1.91–1.58(m,4H),1.38–1.33(m,1H),1.23–1.13(m,1H)ppm.
LC-MS(m/z):508(M+1)。
实施例58
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000080
2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(500mg，1.58mmol)，N-甲基-N-苯基甘氨酸(420mg，2.07mmol)和DIPEA(0.9mL，4.0mmol)溶于DMF(25mL)中，加入HOBT(250mg，1.85mmol)和EDCI(500mg，2.62mmol)，室温反应过夜。反应液倒入300mL水中，调节pH值至8-10，析出固体，过滤获得粗品，用DCM/MeOH＝20/1过柱纯化获得1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(苯基)氨基)乙基-1-酮(390mg，收率57.7％)。
LC-MS(m/z):429(M+1)。
实施例59
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000081
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(苯基)氨基)乙基-1-酮(245mg,63％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(苯基)氨基)乙基-1-酮(390mg,0.91mmol)按照实施例7中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d6)δ7.97(s,0.8H),7.94(s,0.2H),7.63～7.51(m,2H),7.41(t,J＝7.5Hz,1H),7.30(t,J＝7.5Hz,1H),7.17-7.11(m,3H),6.62-6.58(m,3H),5.15(d,J＝5.4Hz,0.16H),5.04(d,J＝5.4Hz,0.84H),4.37(t,J＝13.0Hz,1H),4.23(s,2H),3.92(t,J＝14.3Hz,1H),3.73-3.72(m,1H),3.00-2.93(m,4H),2.47-2.44(m,1H),2.22～2.06(m, 1H),2.01-1.87(m,1H),1.82～1.79(m,1H),1.63～1.56(m,2H),1.31～1.27(m,1H),1.20-1.10(m,1H).
LC-MS(m/z):431(M+1)。
实施例60
N-(3-氯苯基)-N-甲基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000082
往20ml的微波管中加入3-氯-N-甲基苯胺(300mg，2.13mmol)，2-溴乙酸乙酯(426mg，2.55mmol)，二异丙基乙胺(550mg，4.26mmol)和10ml乙腈，混合物在微波下和100摄氏度反应2小时，停止反应，冷却到室温，浓缩，剩余物过柱(展开剂为石油醚：乙酸乙酯＝10:1)得到化合物N-(3-氯苯基)-N-甲基甘氨酸乙酯(460mg，收率95％)。
LC-MS(m/z):228(M+1)。
实施例61
N-(3-氯苯基)-N-甲基甘氨酸的制备
Figure PCTCN2017116914-appb-000083
N-(3-氯苯基)-N-甲基甘氨酸乙酯(460mg，2.02mmol)溶解在甲醇(10ml)和四氢呋喃(10ml)，加入10ml氢氧化锂(1摩尔/升)，混合液室温反应3小时，停止反应，反应液用1摩尔/升的盐酸调pH到6，用乙酸乙酯萃取3遍，合并的有机层用饱和食盐水洗3次，无水硫酸钠干燥，过滤并且旋干，剩余物过柱(展开剂为石油醚：乙酸乙酯＝2:1)得到白色固体N-(3-氯苯基)-N-甲基甘氨酸(260mg，65％)。
LC-MS(m/z):200(M+1)。
实施例62
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-氯苯基)(甲基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000084
往100ml的圆底瓶中加入N-(3-氯苯基)-N-甲基甘氨酸(100mg，0.5mmol)，2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(150mg，0.5mmol)，HATU(285mg，0.75mmol)，二异丙基乙胺(322mg，2.5mmol)和二氯甲烷(30ml)，混合液室温搅拌过夜，停止反应，反应液用水洗涤2遍，无水硫酸钠干燥，过滤，旋干，剩余物过柱(展开剂为二氯甲烷：甲醇＝20:1)得到白色固体1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-氯苯基)(甲基)氨基)乙基-1-酮(100mg,43％)。
LC-MS(m/z):463(M+1)。
实施例63
2-((3-氯苯基)(甲基)氨基)-1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-乙基-1-酮的制备
Figure PCTCN2017116914-appb-000085
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-氯苯 基)(甲基)氨基)乙基-1-酮(100mg，0.2mmol)溶解在10ml的甲醇中，冷却到0℃，加入硼氢化钠(38mg，1mmol)，0度反应2小时，停止反应，反应液倒入100ml的水中，乙酸乙酯萃取三遍，合并的有机相用饱和氯化钠水溶液洗涤两次，无水硫酸钠干燥，过滤，旋干，剩余物过柱(展开剂为二氯甲烷：甲醇＝15:1)，得化合物白色固体2-((3-氯苯基)(甲基)氨基)-1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-乙基-1-酮(40mg，40％)。
1H NMR(400MHz,MeOD)δ8.028(s,0.8H)7.973(s,0.2H),7.628(d,J＝7.6Hz,1H),7.765(d,J＝7.6Hz,0.8H),7.487(d,J＝7.6Hz,0.2H),7.425(t,J＝7.6Hz,1H),7.318-7.363(m,1H),7.168(s,1H),7.108(t,J＝9.6Hz,1H),6.631(s,1H),6.589(t,J＝9.6Hz,1H),5.453-5.546(m,1H),4.536(t,J＝15.2Hz,1H),4.203-4.336(m,2H),4.002(t,J＝15.2Hz,1H),3.725-3.824(m,1H),3.094(t,J＝12.4Hz,1H),3.012(s,3H),2.621(t,J＝12.4Hz,1H),2.069-2.362(m,2H),1.933(t,J＝12Hz,1H),1.622-1.742(m,2H),1.218-1.447(m,2H).
LC-MS(m/z):465(M+1)。
实施例64
(3-氯-4-氟苯基)甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000086
往20ml的微波管中加入3-氯-4-氟苯胺(1000mg，6.89mmol)，2-溴乙酸乙酯(1720mg，10.3mmol)，二异丙基乙胺(2660mg，20.6mmol)和20ml乙腈，混合物在微波，100℃反应2小时，停止反应，冷却到室温，浓缩，剩余物过柱(展开剂为石油醚：乙酸乙酯＝10:1)得到黄色油状物(3-氯-4-氟苯基)甘氨酸乙酯(820mg，51％)。
LC-MS(m/z):232(M+1)。
实施例65
N-(3-氯-4-氟苯基)-N-甲基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000087
(3-氯-4-氟苯基)甘氨酸乙酯(400mg，1.73mmol)溶解在DMF(10ml)中，向其中加入碳酸铯(1680mg，5.19mmol)和碘甲烷(1220mg，8.65mmol)。混合液在50度下搅拌16小时。反应液冷却到室温，用乙酸乙酯(30ml)稀释后用饱和食盐水(30ml)洗4遍。有机相用无水硫酸钠干燥后旋干，剩余物过柱(展开剂为石油醚：乙酸乙酯＝10:1)得到淡黄色固体N-(3-氯-4-氟苯基)-N-甲基甘氨酸乙酯(360mg，85％)。
LC-MS(m/z):246(M+1)。
实施例66
N-(3-氯-4-氟苯基)-N-甲基甘氨酸的制备
Figure PCTCN2017116914-appb-000088
灰白色固体N-(3-氯-4-氟苯基)-N-甲基甘氨酸(170mg,95％)是由N-(3-氯-4-氟苯基)-N-甲基甘氨酸乙酯(200mg,0.816mmol)按照实施例61中的类似步骤制备而成。
LC-MS(m/z):218(M+1)。
实施例67
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-氯-4-氟苯基)(甲基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000089
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-氯-4-氟苯基)(甲基)氨基)乙基-1-酮(130mg,51％)是由N-(3-氯-4-氟苯基)-N-甲基甘氨酸(150mg,0.69mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(150mg,0.53mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):481(M+1)。
实施例68
2-((3-氯-4-氟苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000090
白色固体2-((3-氯-4-氟苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(80mg,61％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-氯-4-氟苯基)(甲基)氨基)乙基-1-酮(130mg,0.27mmol)按照实施例63中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.10(s,1H),7.66～7.49(m,2H),7.43(t,J＝8Hz,1H),7.36(t,J＝6Hz,1H),7.22(s,1H),7.01(t,J＝8Hz,1H),6.69～6.67(m,1H),6.58～6.56(m,1H),5.50～5.47(m,1H),4.56～4.49(t,J＝14Hz,1H),4.29～4.17(m,2H),3.98(t,J＝16Hz,1H),3.82～3.78(m,1H),3.11～3.04(m,1H),2.98(s,3H),2.61(t,J＝14Hz,1H),2.18～2.12(m,2H), 1.95～1.92(m,1H),1.70～1.65(m,2H),1.33～1.22(m,2H).
LC-MS(m/z):483(M+1)。
实施例69
N-(4-氟苯基)-N-甲基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000091
黄色油状物N-(4-氟苯基)-N-甲基甘氨酸乙酯(0.8g,95％)是由4-氟-N-甲基苯胺(500mg,4mmol)和2-溴乙酸乙酯(1.33g,8mmol)按照实施例60中的类似步骤制备而成。
LC-MS(m/z):212(M+1)。
实施例70
N-(4-氟苯基)-N-甲基甘氨酸的制备
Figure PCTCN2017116914-appb-000092
黄色油状物N-(4-氟苯基)-N-甲基甘氨酸(600mg,98％)是由N-(4-氟苯基)-N-甲基甘氨酸乙酯(700mg,3.3mmol)按照实施例61中的类似步骤制备而成。
LC-MS(m/z):184(M+1)。
实施例71
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((4-氟苯基)(甲基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000093
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((4-氟苯基)(甲基)氨基)乙基-1-酮(50mg,24％)是由N-(4-氟苯基)-N-甲基甘氨酸(110mg,0.6mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(130mg,0.46mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):447(M+1)。
实施例72
2-((4-氟苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000094
白色固体2-((4-氟苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(35mg,70％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((4-氟苯基)(甲基)氨基)乙基-1-酮(50mg,0.11mmol)按照实施例63中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.00(s,1H),7.64～7.48(m,2H),7.42(t,J＝6Hz,1H),7.33(t,J＝8Hz,1H),7.15(s,1H),6.90(t,J＝8Hz,2H),6.67～6.65(m,2H),5.50～5.44(m,1H),4.53(t,J＝14Hz,1H),4.26～4.13(m,2H),4.01(t,J＝16Hz,1H),3.80～3.73(m,1H),3.10～3.03(m,1H),2.98(s,3H),2.60(t,J＝14Hz,1H),2.21～2.03(m,2H),1.94～1.89(m,1H), 1.72～1.64(m,2H),1.27～1.19(m,2H).
LC-MS(m/z):449(M+1)。
实施例73
(3,4-二甲基苯基)甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000095
(3,4-二甲基苯基)甘氨酸乙酯(320mg,38％)是由3,4-二甲基苯胺(500mg,4.1mmol)和2-溴乙酸乙酯(897mg,5.4mmol)按照实施例64中的类似步骤制备而成。
LC-MS(m/z):208(M+1)。
实施例74
N-(3,4-二甲基苯基)-N-甲基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000096
白色固体N-(3,4-二苯基)-N-甲基甘氨酸乙酯(130mg,40％)是由(3,4-二甲基苯基)甘氨酸乙酯(300mg,1.45mmol)和碘甲烷(1.02g,7.25mmol)按照实施例65中的类似步骤制备而成。
LC-MS(m/z):222(M+1)。
实施例75
N-(3,4-二甲基苯基)-N-甲基甘氨酸的制备
Figure PCTCN2017116914-appb-000097
白色固体N-(3,4-二甲基苯基)-N-甲基甘氨酸(100mg,88％)是由N-(3,4-二甲基苯基)-N-甲基甘氨酸乙酯(130mg,0.59mmol)按照实施例66中的类似步骤制备而成。
LC-MS(m/z):194(M+1)。
实施例76
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3,4-二甲基苯基)(甲基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000098
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3,4-二甲基苯基)(甲基)氨基)乙基-1-酮(130mg,50％)是由N-(3,4-二甲基苯基)-N-甲基甘氨酸(100mg,0.65mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(160mg,0.5mmol)按照实施例67中的类似步骤制备而成。
LC-MS(m/z):457(M+1)。
实施例77
2-((3,4-二甲基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000099
白色固体2-((3,4-二甲基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(80mg,60％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3,4-二甲基苯基)(甲基)氨基)乙基-1-酮(130mg,0.29mmol)按照实施例68中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.00(s,1H),7.64～7.55(m,2H),7.42(t,J＝6Hz,1H),7.33(t,J＝8Hz,1H),7.15(s,1H),6.90(d,J＝4Hz,1H),6.51(s,1H),6.44(d,J＝4Hz,1H),5.45～5.42(m,1H),4.54(t,J＝14Hz,1H),4.18～4.06(m,3H),3.80～3.77(m,1H),3.09～3.02(m,1H),2.95(s,3H),2.59(t,J＝12Hz,1H),2.18～2.07(m,8H),1.91～1.88(m,1H),1.70～1.63(m,2H),1.37～1.22(m,2H).
LC-MS(m/z):459(M+1)。
实施例78
N-乙基-N-苯基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000100
N-乙基-N-苯基甘氨酸乙酯(3.2g,85％)是由N-乙基苯胺(2.0g,16.5mmol)和2-溴乙酸乙酯(4.11g,24.8mmol)按照实施例60中的类似步骤制备而成。
LC-MS(m/z):208(M+1)。
实施例79
N-乙基-N-苯基甘氨酸的制备
Figure PCTCN2017116914-appb-000101
白色固体N-乙基-N-苯基甘氨酸(90mg,粗品)是由N-乙基-N-苯基甘氨酸乙酯(100mg，0.452mmol)按照实施例61中的类似步骤制备而成。
LC-MS(m/z):180(M+1)。
实施例80
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(乙基(苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000102
褐色固体1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(乙基(苯基)氨基)乙基-1-酮(70mg,粗品)是由N-乙基-N-苯基甘氨酸(79mg,0.442mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(70mg,0.221mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):443(M+1)。
实施例81
2-(乙基(苯基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000103
白色固体2-(乙基(苯基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(15mg,21.3％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(乙基(苯基)氨基)乙基-1-酮(70mg,0.158mmol)按照实施例63中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d6)δ7.95(s,1H),7.61～7.57(m,2H),7.39(t,J＝12Hz,1H),7.29(t,J＝12Hz,1H),7.15～7.08(m,3H),6.56～6.52(m,3H),5.41～5.38(m,1H),5.02(s,1H),4.41～4.36(m,1H),4.13(s,2H),3.98～3.90(m,1H),3.71～3.70(m,1H),3.38(s,1H),2.96～2.93(m,1H),2.47～2.45(m,1H),2.08～2.06(m,1H),1.91～1.88(m,1H),1.80～1.78(m,1H),1.58～1.53(m,2H),1.29～1.22(m,1H),1.13～1.07(m,4H).
LC-MS(m/z):445(M+1)。
实施例82
N-(叔丁氧基羰基)-N-苯基甘氨酸的制备
Figure PCTCN2017116914-appb-000104
将苯基甘氨酸(1.51g，10mmol),(Boc)2O(3.27g,15mmol)和碳酸钠(2.12g，20mmol)溶于甲醇(50mL)和水(20mmol)中，补加碳酸钠(1.06g，10mmol)使pH值在9-10之间，室温反应过夜。反应完全后，反应液中加水(40mL)稀释，盐酸调pH值1-2，用二氯甲烷(40mL*2)萃取,水洗(20mL*2),有机相减压浓缩，获得棕褐色液体N-(叔丁氧基羰基)-N-苯基甘氨酸(2.49g，收率98％)。
LC-MS(m/z):252(M+1)。
实施例83
(2-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-氧基乙基)(苯基)甲酸叔丁酯的制备
Figure PCTCN2017116914-appb-000105
(2-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-氧基乙基)(苯基)甲酸叔丁酯(900mg,57％)是由N-(叔丁氧基羰基)-N-苯基甘氨酸(1.0g,4mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(837mg,2.64mmol)按照实施例58中的类似步骤制备而成。
LC-MS(m/z):515(M+1)。
实施例84
2-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-氧基乙基)(苯基)甲酸叔丁酯的制备
Figure PCTCN2017116914-appb-000106
(2-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-氧基乙基)(苯基)甲酸叔丁酯(600mg,67％)是由(2-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-氧基乙基)(苯基)甲酸叔丁酯(900mg,1.75mmol)按照实施例7中的类似步骤制备而成。
LC-MS(m/z):517(M+1)。
实施例85
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(苯基氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000107
将(2-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-氧基乙基)(苯基)甲酸叔丁酯(600mg，1.45mmol)溶于二氯甲烷(20mL)中，加入浓盐酸(2mL)室温反应3小时。减压浓缩得到残留物，倒入水(300mL)中，调pH值至8-9，析出固体，过滤获得固体用DCM/MeOH＝10/1过柱纯化得到1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(苯基氨基)乙基-1-酮(225mg，收率46.5％)。1H NMR(400MHz,DMSO-d6)δ7.96(s,0.8H),7.92(s,0.2H),7.63～7.51(m,2H),7.40(t,J＝7.5Hz,1H),7.29(t,J＝7.5Hz,1H),7.16(s,0.2H),7.14(s,0.8H),7.08(t,J＝7.6Hz,2H),6.66(d,J＝7.9Hz,2H),6.57(t,J＝7.1Hz,1H),5.51(s,br,1H),5.41(t,J＝6.5Hz,1H),5.14(d,J＝5.8Hz,0.16H),5.03(d,J＝5.8Hz,0.84H),4.45(t,J＝13.5Hz,1H),3.97(t,J＝14.5Hz,1H),3.88(s,2H),3.71(s,br,1H),2.96(t,J＝12.9Hz,1H),2.12～2.05(m,1H),1.93-1.80(m,2H),1.64～1.54(m,2H),1.32～1.15(m,2H).
LC-MS(m/z):417(M+1)。
实施例86
N-甲基-N-(4-三氟甲基苯基)甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000108
黄色油状物N-甲基-N-(4-三氟甲基苯基)甘氨酸乙酯(630mg,84％)是由N-甲基-4-三氟甲基苯胺(500mg,2.86mmol)和2-溴乙酸乙酯(0.96g,5.72mmol)按照实施例60中的类似步骤制备而成。
LC-MS(m/z):262(M+1)。
实施例87
N-甲基-N-(4-三氟甲基苯基)甘氨酸的制备
Figure PCTCN2017116914-appb-000109
淡黄色油状物N-甲基-N-(4-三氟甲基苯基)甘氨酸(470mg,87％)是由N-甲基-N-(4-三氟甲基苯基)甘氨酸乙酯(600mg,2.3mmol)按照实施例61中的类似步骤制备而成。
LC-MS(m/z):234(M+1)。
实施例88
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(4-三氟甲基苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000110
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(4-三氟甲基苯基)氨基)乙基-1-酮(60mg,26％)是由N-甲基-N-(4-三氟甲基苯基)甘氨酸(140mg，0.6mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(130mg,0.46mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):497(M+1)。
实施例89
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(4-三氟甲基苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000111
白色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(4-三氟甲基苯基)氨基)乙基-1-酮(35mg,58％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(4-三氟甲基苯基)氨基)乙基-1-酮(60mg,0.12mmol)按照实施例63中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.28(s,1H),7.69～7.58(m,2H),7.47～7.36(m,4H),7.30～7.28(m,1H),6.73(d,J＝8Hz,2H),5.63～5.53(m,1H),4.52(t,J＝14Hz,1H),4.40～4.28(m,2H),4.00(t,J＝18Hz,1H),3.80～3.69(m,1H),3.13～3.02(m,4H),2.61(t,J＝12Hz,1H),2.20～2.15(m,2H),1.94～1.88(m,1H),1.75～1.58(m,2H),1.45～1.37(m,2H).
LC-MS(m/z):499(M+1)。
实施例90
(3-三氟甲基苯基)甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000112
黄色油状物(3-三氟甲基苯基)甘氨酸乙酯(850mg,55％)是由3-三氟甲基苯胺(1g,6.2mmol)和2-溴乙酸乙酯(1.55g,9.3mmol)按照实施例64中的类似步骤制备而成。
LC-MS(m/z):248(M+1)。
实施例91
N-甲基-N-(3-三氟甲基苯基)甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000113
白色固体N-甲基-N-(3-三氟甲基苯基)甘氨酸乙酯(430mg,81％)是由N-甲基-3-三氟甲基苯胺(500mg,2.02mmol)和碘甲烷(1.43g,10.1mmol)按照实施例65中的类似步骤制备而成。
LC-MS(m/z):262(M+1)。
实施例92
N-甲基-N-(3-三氟甲基苯基)甘氨酸的制备
Figure PCTCN2017116914-appb-000114
黄色油状物N-甲基-N-(3-三氟甲基苯基)甘氨酸(320mg,90％)是由N-甲基-N-(3-三氟甲基苯基)甘氨酸乙酯(400mg,1.5mmol)按照实施例66中的类似步骤制备而成。
LC-MS(m/z):234(M+1)。
实施例93
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(3-三氟甲基苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000115
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(3-三氟甲基苯基)氨基)乙基-1-酮(60mg,24％)是由N-甲基-N-(3-三氟甲基苯基)甘氨酸(151mg,0.65mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(140mg,0.5mmol)按照实施例58中的类似步骤制备而成。
LC-MS(m/z):497(M+1)。
实施例94
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(3-三氟甲基苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000116
白色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(3-三氟甲基苯基)氨基)乙基-1-酮(35mg,58％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(3-三氟甲基苯基)氨基)乙基-1-酮(60mg,0.12mmol)按照实施例7中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.02(s,1H),7.64～7.56(m,2H),7.44～7.40(m,1H),7.35～7.29(m,2H),7.16(s,1H),6.91～6.86(m,3H), 5.54～5.45(m,1H),4.53(t,J＝20Hz,1H),4.40～4.26(m,2H),4.02(t,J＝18Hz,1H),3.82～3.77(m,1H),3.13～3.06(m,4H),2.62(t,J＝12Hz,1H),2.20～2.15(m,2H),1.98～1.90(m,1H),1.75～1.61(m,2H),1.43～1.34(m,2H).
LC-MS(m/z):499(M+1)。
实施例95
N-(4-腈基苯基)-N-甲基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000117
黄色油状物-N-(4-腈基苯基)-N-甲基甘氨酸乙酯(260mg,32％)是由N-甲基-4-腈基苯胺(500mg,3.79mmol)和2-溴乙酸乙酯(759mg,4.55mmol)按照实施例60中的类似步骤制备而成。
LC-MS(m/z):219(M+1)。
实施例96
N-(4-腈基苯基)-N-甲基甘氨酸的制备
Figure PCTCN2017116914-appb-000118
黄色油状物N-(4-腈基苯基)-N-甲基甘氨酸(250mg,92％)是由N-(4-腈基苯基)-N-甲基甘氨酸乙酯(310mg,1.42mmol)按照实施例61中的类似步骤制备而成。
LC-MS(m/z)191:(M+1)。
实施例97
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((4-腈基苯基)(甲基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000119
白色固体1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((4-腈基苯基)(甲基)氨基)乙基-1-酮(100mg,63％)是由N-(4-腈基苯基)-N-甲基甘氨酸(74mg,0.39mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(100mg,0.36mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):454(M+1)。
实施例98
2-((4-腈基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000120
白色固体2-((4-腈基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(45mg,45％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((4-腈基苯基)(甲基)氨基)乙基-1-酮(100mg,0.22mmol)按照实施例63中的类似步骤制备而成。
1HNMR(400MHz,CD3OD)δ8.006(d,1H)7.622(d,J＝7.6Hz,1H),7.563(d,J＝7.6,1H),7.403-7.504(m,3H),7.335(t,J＝7.6Hz,1H),7.157(d,1H),6.703-6.731(m,2H),5.448-5.537(m,1H),4.528(t,J＝16Hz,1H),4.325-4.439(m,2H),3.974(t,J＝16Hz,1H),3.751-3.819(m,1H),3.078-3.134(m,4H),2.624(t,J＝13.6Hz,1H),2.067-2.367(m,2H),1.939(t,J＝13.6Hz,1H),1.610-1.802(m,2H),1.212-1.482(m,2H).
LC-MS(m/z):456(M+1)。
实施例99
N-(3-甲氧基苯基)-N-甲基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000121
N-(3-甲氧基苯基)-N-甲基甘氨酸乙酯(400mg,49％)是由3-甲氧基-N-甲基苯胺(500mg,3.65mmol)和2-溴乙酸乙酯(731mg,4.38mmol)按照实施例60中的类似步骤制备而成。
LC-MS(m/z):224(M+1)。
实施例100
N-(3-甲氧基苯基)-N-甲基甘氨酸的制备
Figure PCTCN2017116914-appb-000122
N-(3-甲氧基苯基)-N-甲基甘氨酸(300mg,85％)是由N-(3-甲氧基苯基)-N-甲基甘氨酸乙酯(400mg,1.8mmol)按照实施例61中的类似步骤制备而成。
LC-MS(m/z):196(M+1)。
实施例101
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-甲氧基苯基)(甲基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000123
白色固体1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-甲氧基苯基)(甲基)氨基)乙基-1-酮(100mg,61％)是由N-(3-甲氧基苯基)-N-甲基甘氨酸(76mg,0.39mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(100mg,0.36mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):459(M+1)。
实施例102
2-((3-甲氧基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000124
白色固体2-((3-甲氧基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(55mg,55％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-((3-甲氧基苯基)(甲基)氨基)乙基-1-酮(100mg,0.22mmol)按照实施例63中的类似步 骤制备而成。
1HNMR(400MHz,CD3OD)δ8.032(s,1H),7.629(d,J＝7.6Hz,1H),7.580(d,J＝7.6Hz,1H),7.427(t,J＝7.6Hz,1H),7.339(t,J＝7.6Hz,1H),7.172(s,1H),7.056(t,J＝8Hz,1H),6.272(t,J＝9.6Hz,2H),6.194(s,1H),5.445-5.541(m,1H),4.543(t,J＝15.2Hz,1H),4.141-4.290(m,2H),4.023(t,J＝15.2Hz,1H),3.764-3.811(m,1H),3.723(s,3H),3.076(t,J＝12.8Hz,1H),3.002(s,3H),2.608(t,J＝12.8Hz,1H),2.032-2.348(m,2H),1.911(t,J＝11.2Hz,1H),1.615-1.783(m,2H),1.192-1.423(m,2H).
ESI LC-MS(m/z):461(M+1)。
实施例103
N-甲基-N-(4-硝基苯基)甘氨酸的制备
Figure PCTCN2017116914-appb-000125
0℃下将NaH(60％in mineral oil,960mg,24mmol)加入N-甲基-4-硝基苯胺(3g,20mmol)的四氢呋喃溶液中(50ml)。0℃下搅拌0.5小时后向此反应液加入2-溴乙酸乙酯(3.65g,22mmol)。在室温下反应2小时。然后加入NaOH溶液(1N,40ml,40mmol)，继续反应1小时。停止反应，反应液倒入100ml水中，用乙酸乙酯萃取两次(100ml X 2)，合并的有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，旋干，剩余物过柱(展开剂为二氯甲烷：甲醇＝30：1)得到黄色固体N-(4-硝基苯基)-N-甲基甘氨酸(1.2g，28.6％)。
LC-MS(m/z):211(M+1)。
实施例104
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(4-硝基苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000126
黄色固体1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(4-硝基苯基)氨基)乙基-1-酮(70mg,55％)是由N-甲基-N-(4-硝基苯基)甘氨酸(50mg,0.23mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(60mg,0.21mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):474(M+1)。
实施例105
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(4-硝基苯基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000127
黄色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(4-硝基苯基)氨基)乙基-1-酮(30mg,42％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(4-硝基苯基)氨基)乙基-1-酮(70mg,0.15mmol)按照实施例63中的类似步骤制备而成。
1HNMR(400MHz,MeOD)δ8.079(d,J＝9.6Hz,2H)8.015(s,1H),7.627(d,J＝7.6Hz,1H),7.590(d,J＝7.6Hz,1H),7.426(t,J＝7.6Hz,1H),7.340(t,J＝7.6Hz,1H),7.162(s,1H),6.702(d,J＝9.6Hz,2H),4.462-5.556(m,1H),4.407-4.577(m,2H),3.982(t,J＝13.2Hz,1H),3.730-3.830(m,1H),3.091-3.150(m,4H),2.642(t,J＝13.2Hz,1H),2.110-2.200(m,1H),1.949(t, J＝15.6Hz,1H),1.614-1.762(m,2H),1.278-1.461(m,3H).
LC-MS(m/z):476(M+1)。
实施例106
N-甲基-N-(吡啶-2-基)甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000128
黄色油状物N-甲基-N-(吡啶-2-基)甘氨酸乙酯(650mg,60％)是由N-甲基-吡啶-2-胺(600mg,5.5mmol)和2-溴乙酸乙酯(1.39g,8.3mmol)按照实施例60中的类似步骤制备而成。
LC-MS(m/z):195(M+1)。
实施例107
N-甲基-N-(吡啶-2-基)甘氨酸的制备
Figure PCTCN2017116914-appb-000129
黄色油状物N-甲基-N-(吡啶-2-基)甘氨酸(160mg,93％)是由N-甲基-N-(吡啶-2-基)甘氨酸乙酯(200mg,1.03mmol)按照实施例61中的类似步骤制备而成。
LC-MS(m/z):167(M+1)。
实施例108
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(吡啶-2-基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000130
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(吡啶-2-基)氨基)乙基-1-酮(70mg,29％)是由N-甲基-N-(吡啶-2-基)甘氨酸(122mg,0.74mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(160mg,0.57mmol)按照实施例58中的类似步骤制备而成。
LC-MS(m/z):430(M+1)。
实施例109
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(吡啶-2-基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000131
白色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(吡啶-2-基)氨基)乙基-1-酮(19mg,27％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(吡啶-2-基)乙基-1-酮(70mg,0.16mmol)按照实施例7中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.02～7.97(m,2H),7.64～7.62(d,J＝8Hz,1H),7.58～7.50(m,2H),7.42(t,J＝16Hz,1H),7.33(t,J＝16Hz,1H),7.16(s,1H),6.66(d,J＝8Hz,1H),6.60(t,J＝12Hz,1H),5.48～5.45(m,1H),4.56～4.41(m,3H),4.06～3.98(m,1H),3.87～3.78(m,1H),3.12～3.07(m,4H),2.60(t,J＝12Hz,1H),2.18～2.10(m,2H),1.94～1.88(m,1H),1.68～1.59(m,2H),1.43～1.35(m,2H).
LC-MS(m/z):432(M+1)。
实施例110
喹啉-6-基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000132
往100ml的梨型烧瓶中中加入6-氨基喹啉(2000mg，13.9mmol)，2-溴乙酸乙酯(3479mg，20.8mmol)，二异丙基乙胺(5375mg，41.6mmol)和30ml乙腈，混合物在50摄氏度反应16小时，停止反应，冷却到室温，浓缩，剩余物过柱(展开剂为石油醚：乙酸乙酯＝10:1)得到喹啉-6-基甘氨酸乙酯(1200mg，收率37％)。
LC-MS(m/z):231(M+1)。
实施例111
N-甲基-N-(喹啉-6-基)甘氨酸的制备
Figure PCTCN2017116914-appb-000133
喹啉-6-基甘氨酸乙酯(1200mg，5.21mmol)溶解在甲醇(20ml)中，向其中加入甲醛(1513mg，52.1mmol)，反应液在室温下搅拌2小时，再向其中加入氰基硼氢化钠(1643mg，26.1mmol)。混合液在室温下搅拌16小时。50ml水加入到反应液中，再用乙酸乙酯(30ml)萃取一遍，取水相用1摩尔每升的盐酸水溶液调节pH值到5～6，再用二氯甲烷/异丙醇(v/v＝3/1,30ml)萃取三遍。合并的有机相用无水硫酸钠干燥后旋干得到黄色油状物N-甲基-N-(喹啉-6-基)甘氨酸(310mg，28％)。
LC-MS(m/z):217(M+1)。
实施例112
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(喹啉-6-基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000134
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(喹啉-6-基)氨基)乙基-1-酮(45mg,26％)是由N-甲基-N-(喹啉-6-基)甘氨酸(100mg,0.46mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(100mg,0.36mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):480(M+1)。
实施例113
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(喹啉-6-基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000135
浅黄色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(喹啉-6-基)氨基)乙基-1-酮(15mg,33％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(喹啉-6-基)乙基-1-酮(45mg,0.09mmol)按照实施例63中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.50(s,1H),8.11(d,J＝8Hz,1H),8.04(s,1H),7.84(d,J＝8Hz,1H),7.64(d,J＝8Hz,1H),7.58(d,J＝8Hz,1H),7.45～7.32(m,4H),7.17(s,1H),6.91(s,1H),5.49～5.46(m,1H),4.63～4.42(m,3H),4.09～4.02(m,1H),3.82～3.81(m,1H),3.19～3.09(m, 4H),2.64(t,J＝14Hz,1H),2.22～2.10(m,2H),1.99～1.90(m,1H),1.78～1.62(m,2H),1.47～1.37(m,2H)。
LC-MS(m/z):482(M+1)。
实施例114
N-甲基-9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-胺的制备
Figure PCTCN2017116914-appb-000136
向100ml梨形烧瓶中加入6-氯-9-(四氢-2H-吡喃-2-基)-9H-嘌呤(1500mg，6.3mmol)和甲胺醇溶液(30ml)。反应液在75度下搅拌5小时。旋干后得到黄色油状物N-甲基-9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-胺(900mg，61％)。
LC-MS(m/z):234(M+1)。
实施例115
N-甲基-N-(9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-基)甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000137
将N-甲基-9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-胺(900mg，3.86mmol)和DMF(10ml)加入圆底烧瓶中，在0℃下加入氢化钠(60％含量)(310mg，7.72mmol)。反应液在35度下搅拌1小时。再向反应液中加入2-溴乙酸乙酯(967mg，5.79mmol)。反应液在35度下搅拌3小时。加入20ml水进行粹灭。之后加入乙酸乙酯(50ml)，再用饱和食盐水(40ml)洗4遍。有机相用无水硫酸钠干燥，过滤并且旋干，剩余物过柱(展开剂为二氯甲烷：甲醇＝20:1)得到白色固体N-甲基-N-(9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-基)甘氨酸乙酯(1100mg，收率：89％)。
LC-MS(m/z):320(M+1)。
实施例116
N-甲基-N-(9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-基)甘氨酸的制备
Figure PCTCN2017116914-appb-000138
N-甲基-N-(9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-基)甘氨酸乙酯(550mg，1.72mmol)溶解在甲醇(10ml)和四氢呋喃(10ml)，加入10ml氢氧化锂(1摩尔/升)，混合液室温反应3小时，停止反应，反应液用1摩尔/升的盐酸调pH到6，用乙酸乙酯萃取3遍，合并的有机层用饱和食盐水洗3次，无水硫酸钠干燥，过滤并且旋干得到白色固体N-甲基-N-(9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-基)甘氨酸(430mg，86％)。
LC-MS(m/z):292(M+1)。
实施例117
N-甲基-N-(9H-嘌呤-6-基)甘氨酸的制备
Figure PCTCN2017116914-appb-000139
N-甲基-N-(9-(四氢-2H-吡喃-2-基)-9H-嘌呤-6-基)甘氨酸(170毫克，0.58毫摩尔)溶解在盐酸/二氧六环溶液(10毫升)和DMF(0.5毫升)中。反应液在室温下搅拌1小时。旋干得到黄色油状物N-甲基-N-(9H-嘌呤-6-基)甘氨酸(130毫克，92％)。LC-MS(m/z)208(M-HCl+1)。
实施例118
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(7H-嘌呤-6-基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000140
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(7H-嘌呤-6-基)氨基)乙基-1-酮(60mg,24％)是由N-甲基-N-(9H-嘌呤-6-基)甘氨酸(130mg,0.69mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(150mg,0.54mmol)按照实施例62中的类似步骤制备而成。
LC-MS(m/z):471(M+1)。
实施例119
1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(7H-嘌呤-6-基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000141
白色固体1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(7H-嘌呤-6-基)氨基)乙基-1-酮(13mg,22％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(甲基(7H-嘌呤-6-基)乙基-1-酮(60mg,0.13mmol)按照实施例63中的类似步骤制备而成。
1H NMR(400MHz,CD3OD)δ8.21(s,1H),8.03～7.90(m,2H), 7.64～7.57(m,2H),7.44～7.32(m,2H),7.17(s,1H),5.49～5.46(m,1H),4.51(t,J＝14Hz,1H),4.08～4.00(m,1H),3.82～3.77(m,1H),3.50～3.33(m,4H),3.17～3.11(m,1H),2.66～2.60(m,1H),2.23～2.10(m,2H),1.96～1.86(m,4H),1.33～1.25(m,2H)。
LC-MS(m/z):473(M+1)。
实施例120
二苯并[b,d]呋喃-3-基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000142
黄色油状物二苯并[b,d]呋喃-3-基甘氨酸乙酯(570mg,65％)是由3-氨基二苯并[b,d]呋喃(600mg,3.3mmol)和2-溴乙酸乙酯(821mg,4.9mmol)按照实施例64中的类似步骤制备而成。
LC-MS(m/z):270(M+1)。
实施例121
N-(二苯并[b,d]呋喃-3-基)-N-甲基甘氨酸乙酯的制备
Figure PCTCN2017116914-appb-000143
灰白色固体N-(二苯并[b,d]呋喃-3-基)-N-甲基甘氨酸乙酯(350mg,83％)是由二苯并[b,d]呋喃-3-基甘氨酸乙酯(400mg,1.48mmol)和碘甲烷(1.05g,7.43mmol)按照实施例65中的类似步骤制备而成。
LC-MS(m/z):284(M+1)。
实施例122
N-(二苯并[b,d]呋喃-3-基)-N-甲基甘氨酸的制备
Figure PCTCN2017116914-appb-000144
淡黄色固体N-(二苯并[b,d]呋喃-3-基)-N-甲基甘氨酸(200mg,92％)是由N-(二苯并[b,d]呋喃-3-基)-N-甲基甘氨酸乙酯(240mg,0.85)按照实施例66中的类似步骤制备而成。
LC-MS(m/z):256(M+1)。
实施例123
1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(二苯并[b,d]呋喃-3-基(甲基)氨基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000145
黄色油状物1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(二苯并[b,d]呋喃-3-基(甲基)氨基)乙基-1-酮(150mg,45％)是由N-(二苯并[b,d]呋喃-3-基)-N-甲基甘氨酸(190mg,0.83mmol)和2-(5H-咪唑并[5,1-a])异吲哚-5-基)-1-(哌啶-4-基)乙基-1-酮盐酸盐(180mg,0.64mmol)按照实施例67中的类似步骤制备而成。
LC-MS(m/z):519(M+1)。
实施例124
2-(二苯并[b,d]呋喃-3-基(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮的制备
Figure PCTCN2017116914-appb-000146
白色固体2-(二苯并[b,d]呋喃-3-基(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮(70mg,46％)是由1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙酰基)哌啶-1-基)-2-(二苯并[b,d]呋喃-3-基(甲基)氨基)乙基-1-酮(150mg,0.29mmol)按照实施例68中的类似步骤制备而成。
1H NMR(400MHz,DMSO-d6)δ8.13(s,1H),7.88(d,J＝8Hz,1H),7.79(d,J＝8Hz,1H),7.66～7.59(m,2H),7.52(d,J＝8Hz,1H),7.41(t,J＝16Hz,1H),7.33～7.27(m,3H),7.22(s,1H),6.81(s,1H),6.71(d,J＝12Hz,1H),5.48～5.44(m,1H),5.09～5.06(m,1H),4.38～4.31(m,3H),3.96～3.89(m,1H),3.73～3.72(m,1H),3.02～2.95(m,4H),2.14～2.06(m,1H),1.96～1.91(m,1H),1.83～1.77(m,1H),1.65～1.55(m,2H),1.36～1.25(m,3H)。
LC-MS(m/z):521(M+1)。
体外生物学评价
本检测方法用于本发明所述化合物的体外生物学活性评价，包括体外酶学活性评价和细胞水平酶学活性评价。
本检测的目的在于综合评价不同化合物对人吲哚胺2,3-双加氧酶(IDO)的体外酶学抑制活性和对细胞模型的酶学抑制活性。
实施例125体外酶学活性检测
实验原理
体外IDO酶学活性检测的基本原理是利用IDO酶在体外酶催化反应体系中将底物L-色氨酸代谢生成产物犬尿氨酸，后者能够与对二甲氨基 苯甲醛的冰乙酸溶液发生颜色反应呈黄色，并在酶标仪492nm处测定吸光度值。通过与已知浓度的犬尿氨酸标准品曲线对比，计算样品中产物犬尿氨酸的浓度。当加入不同测试化合物时，其对IDO酶活性的抑制体现为产物犬尿氨酸的减少及颜色反应的改变。
实验材料与试剂
重组人IDO酶(IDO1酶)购自美国BPS Bioscience公司。检测试剂如L-色氨酸(生工，A601911-0050)、抗坏血酸(生工，SB0830-100g)、过氧化氢酶(生工，54J15066)、亚甲基蓝(天津基准化学试剂有限公司)、犬尿氨酸(sigma，K8625-100MG)、三氯乙酸(生工，A600968-0250)、对二甲氨基苯甲醛(天津市大茂化学试剂厂)。
实验主要过程
实验主要流程如下：
(1)实验准备：按照要求配制2×IDO酶反应缓冲液(工作液)：0.1M磷酸钾缓冲液(pH＝6.5)、400μM L-色氨酸、40mM抗坏血酸、2000U/ml过氧化氢酶、40μM亚甲基蓝；用0.1M磷酸钾缓冲液(pH＝6.5)将测试化合物稀释成不同浓度梯度的工作液(化合物最高浓度均为10μM)。30％(w/v)三氯乙酸溶液。2％对二甲氨基苯甲醛的冰乙酸溶液。
(2)酶学反应体系200μL，包括100μL测试化合物、100μL酶反应缓冲液和0.4μL重组人IDO酶溶液(终浓度为35nM)或1μL重组人IDO酶溶液(终浓度为70nM)。混匀后，IDO酶反应体系在37℃反应30min。
(3)检测反应同时设置有对照反应，包括未添加测试化合物的0抑制阳性对照和未添加酶的0酶阴性对照。所有检测采用复孔。
(4)酶学反应结束后，加入40μL预先配制的30％(w/v)三氯乙酸溶液，65℃反应20min，随后12000rpm离心15min。
(5)吸取100μL离心后的上清液加入96孔平底板中，再加入等体积2％对二甲氨基苯甲醛的冰乙酸溶液混匀，室温放置10min。
(6)使用酶标仪(ELX800NB)检测每孔的颜色信号，检测波长为 492nm。
(7)测试化合物的酶学抑制率计算公式：酶活抑制率(％)＝(0抑制阳性对照-化合物检测孔)/(0抑制阳性对照-0酶阴性对照)*100％。另外，针对不同浓度梯度的测试化合物分别计算出酶学抑制率，然后利用IC50计算器计算酶学半抑制浓度(IC50)。
根据上述实验方法，以现有技术中已知的化合物NLG919(CAS:1402836-58-1)为阳性对照化合物，在相同条件下将本发明所述化合物进行体外IDO酶学评价(对IDO1检测，测试化合物浓度为200nM)。数据小结见下表(表1)。
表1本发明代表性化合物抑制IDO的酶学数据
Figure PCTCN2017116914-appb-000147
由上表结果可见，本发明化合物具有良好的IDO酶抑制活性，且多数优于对照化合物NLG919。
根据上述实验方法，以NLG919为阳性对照化合物，将本发明所述化合物进行体外IDO酶学IC50测定。数据小结见下表(表2)，可见本发 明代表性化合物具有更低的体外IDOIC50值。
表2本发明代表性化合物的体外IDO酶学IC50(nM)
实施例 IC50(nM)
7 55
59 159
85 144
NLG919 198
实施例126细胞水平抑制活性检测
IDO酶除了在免疫细胞如髓源性抑制细胞(MDSC)中组成性表达外，许多肿瘤细胞也会上调表达IDO，或经细胞因子如IFN-γ诱导表达。在本发明中，我们利用IFN-γ诱导Hela细胞表达IDO酶(IDO1酶)为模型，检测化合物在细胞水平的IDO酶学抑制活性。
实验原理
HeLa细胞为人宫颈癌细胞系，在人IFN-γ诱导下可上调表达内源性IDO酶。通过在细胞培养液中添加底物L-色氨酸，可以对细胞上清进行酶催化产物犬尿氨酸的检测。实验采用培养的Hela细胞，在人IFN-γ刺激下，与不同浓度的测试化合物共孵育指定时间后，利用酶产物与对二甲氨基苯甲醛的颜色反应方法，检测测试化合物处理对细胞IDO酶活性的影响。
实验材料与试剂
重组人IFN-γ细胞因子购自生工生物公司，细胞培养用无酚红DMEM购自Gibco公司。检测试剂如L-色氨酸(生工，A601911-0050)、犬尿氨酸(sigma，K8625-100MG)、三氯乙酸(生工，A600968-0250)、对二甲氨基苯甲醛(天津市大茂化学试剂厂)、细胞培养用96孔平底板(CORNING,costar 3599)。
实验过程
按常规细胞培养实验操作流程，在96孔平底板中进行。
(1)Hela细胞按照适当的浓度(约20000个/每孔)接种到96孔培养板中，经过夜贴壁后，换用含200μM L-色氨酸的无酚红DMEM培养基，同时加入细胞因子人IFN-γ50ng/ml和不同浓度梯度的测试化合物(最高终浓度25μM)，同时设置溶剂对照(DMSO)以及无细胞因子和L-色氨酸的阴性对照孔，按照3次重复孔设置。细胞继续培养48小时后进行检测。
(2)吸出培养孔中的上清200μL，加入40μL预先配制的30％(w/v)三氯乙酸溶液，65℃反应20min。随后12000rpm离心15min。
(3)吸取100μL离心后的上清液加入96孔平底板中，再加入等体积2％对二甲氨基苯甲醛的冰乙酸溶液混匀，室温放置10min。
(4)使用酶标仪(ELX800NB)检测每孔的颜色信号，检测波长为492nm。
(5)测试化合物的细胞水平酶学抑制率计算公式：酶活抑制率(％)＝(0抑制阳性对照-化合物检测孔)/(0抑制阳性对照–阴性对照)*100％。另外，针对不同浓度梯度的测试化合物分别计算出细胞酶活抑制率，然后利用EC50计算器计算细胞酶活半抑制浓度(EC50)。
根据上述实验方法，以NLG919为阳性对照化合物，将本发明所述化合物进行细胞学水平IDO酶学评价(测试化合物浓度为1μM)。数据小结见下表(表3)。由表3结果可见，本发明化合物具有良好的IDO抑制活性，且多数优于对照化合物NLG919。
表3本发明代表性化合物的人IDO细胞水平抑制率数据
Figure PCTCN2017116914-appb-000148
Figure PCTCN2017116914-appb-000149
根据上述实验方法，以NLG919为阳性对照化合物，将本发明所述化合物进行IDO细胞水平EC50测定。数据小结见下表(表4)。
表4本发明代表性化合物IDO细胞水平EC50(nM)
实施例 IC50(nM)
7 154
59 169
85 312
NLG919 430
根据上述结果可见，本发明的代表性化合物均具有较低的IDO细胞水平EC50值，优于对照化合物NLG919。
。

Claims (19)

  1. 一种通式I的化合物，
    Figure PCTCN2017116914-appb-100001
    或其可药用盐，
    其中，
    R1为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基、C1-6卤代烷基、C1-6烷基羰基和C1-6烷基氨基；
    R2为羟基或氨基；
    R3为一个或多个取代基，选自氢、羟基、氰基、卤素、C1-6烷基、C1-6烷氧基、C1-6卤代烷基、C1-6烷基羰基和C1-6烷基氨基；
    X为NR4或O；
    其中，R4为氢或C1-6烷基；
    环A为任选经取代的选自以下的基团：苯基；3元到7元饱和或部分不饱和碳环；8元到10元饱和、部分不饱和或芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；具有1到3个独立地选自氮、氧或硫的杂原子的4元到7元饱和或部分不饱和杂环；具有1到5个独立地选自氮、氧或硫的杂原子的7元到10元饱和或部分不饱和杂环双环或三环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳环双环或三环。
  2. 根据权利要求1所述的通式I化合物，其中：
    R1为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素和C1-6卤代烷基；
    R2为羟基；
    R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
    X为NR4；
    其中，R4为氢、甲基或乙基；
    环A为以下的基团：苯基；3元到6元饱和碳环；8元到10元芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳环；具有1到5个独立地选自氮、氧或硫的杂原子的7元到10元饱和或部分不饱和杂环双环或三环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳环双环或三环。
  3. 根据权利要求1所述的通式I化合物，其中：
    R1为一个或多个取代基，选自氢、羟基、氰基、硝基和卤素；
    R2为羟基；
    R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
    X为NR4；
    其中，R4为氢或甲基；
    环A为选自以下的基团：苯基；3元到6元饱和碳环；8元到10元芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
  4. 根据权利要求1所述的通式I化合物，其中：
    R1为一个或多个取代基，选自氢、羟基、氰基、硝基；
    R2为羟基；
    R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
    X为NR4；
    其中，R4为氢或甲基；
    环A为选自以下的基团：苯基；8元到10元芳环双环或三环；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
  5. 根据权利要求1所述的通式I化合物，其中：
    R1为一个或多个取代基，选自氢、羟基和氰基；
    R2为羟基；
    R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
    X为NR4；
    其中，R4为氢或甲基；
    环A为选自以下的基团：苯基；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
  6. 根据权利要求1所述的通式I化合物，其中：
    R1为氢；
    R2为羟基；
    R3为一个或多个取代基，选自氢、羟基、氰基、硝基、卤素、C1-6烷基、C1-6烷氧基和C1-6卤代烷基；
    X为NR4；
    其中，R4为氢或甲基；
    环A为选自以下的基团：苯基；具有1到4个独立地选自氮、氧或硫的杂原子的5元到6元单环杂芳基环；或具有1到5个独立地选自氮、氧或硫的杂原子的8元到10元杂芳基环双环或三环。
  7. 根据权利要求1所述的化合物，其中所述化合物选自：
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-苯氧乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-((四氢 呋喃-3-基)氧基)乙基-1-酮；
    2-((9H-咔唑-3-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(哒嗪-4-基氧基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(喹啉-6-基氧基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(喹啉-6-基氧基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(吡咯烷-3-基氧基)乙基-1-酮；
    2-((7H-嘌呤-6-基)氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(苯并[d]恶唑啉-2-基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(3-氯-4-氟苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(3-氯苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(4-氟苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(3,4-二甲基苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(3-三氟甲基苯氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(环己基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶 -1-基)乙基-1-酮；
    2-(二苯并[b,d]呋喃-2-基氧基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(苯基)氨基)乙基-1-酮；
    2-((3-氯苯基)(甲基)氨基)-1-(4-(2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-乙基-1-酮；
    2-((3-氯-4-氟苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-((4-氟苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-((3,4-二甲基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-(乙基(苯基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(苯基氨基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(4-三氟甲基苯基)氨基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(3-三氟甲基苯基)氨基)乙基-1-酮；
    2-((4-腈基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    2-((3-甲氧基苯基)(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(4- 硝基苯基)氨基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(吡啶-2-基)氨基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(喹啉-6-基)氨基)乙基-1-酮；
    1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)-2-(甲基(7H-嘌呤-6-基)氨基)乙基-1-酮；和
    2-(二苯并[b,d]呋喃-3-基(甲基)氨基)-1-(4-(1-羟基-2-(5H-咪唑并[5,1-a]异吲哚-5-基)乙基)哌啶-1-基)乙基-1-酮；
  8. 制备如权利要求1所述的通式I化合物的方法，
    Figure PCTCN2017116914-appb-100002
    包括使式C化合物
    Figure PCTCN2017116914-appb-100003
    在有机溶剂和还原剂作用下反应形成通式I化合物，其中
    R1、R2、R3、X和A如权利要求1中所定义。
  9. 根据权利要求8所述的制备方法，其中所述还原剂选自NaBH4、KBH4、NaBH(OAc)3、KBH(OAc)3和NaBH3CN。
  10. 根据权利要求8或9所述的制备方法，其中所述有机溶剂选自甲醇、乙醇、四氢呋喃。
  11. 式C化合物：
    Figure PCTCN2017116914-appb-100004
    其中，R1、R3、X和A如权利要求1中所定义。
  12. 制备如权利要求11所述的式C化合物的方法，
    包括使式A化合物
    Figure PCTCN2017116914-appb-100005
    与式B化合物
    Figure PCTCN2017116914-appb-100006
    在有机溶剂和偶联试剂的作用下偶联形成式C化合物，其中，R1、R3、X和A如权利要求1中所定义。
  13. 根据权利要求12所述的制备方法，其中所述偶联试剂选自1-乙基 -3-(3-二甲胺丙基)碳二亚胺(EDC)，N,N'-二环己基碳二亚胺(DCC)，N,N'-羰基二咪唑(CDI)和O-(7-氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU)。
  14. 根据权利要求12或13所述的制备方法，其中所述有机溶剂选自苯、甲苯、四氢呋喃、二氧六环、二氯甲烷、氯仿和N,N'-二甲基甲酰胺。
  15. 权利要求11所述的式C化合物作为用于制备权利要求1至7中任一项所述的化合物的中间体的用途。
  16. 一种药物组合物，其包含权利要求1至7中任一项所述的化合物以及任选的药用辅料。
  17. 权利要求1至7中任一项的化合物在制备用作吲哚胺2,3-双加氧酶(IDO)抑制剂的药物中的应用。
  18. 权利要求1至7中任一项的化合物在制备用于治疗或预防癌症、眼部疾病、自身免疫性疾病、心理障碍、抑郁症或焦虑症的药物中的应用。
  19. 权利要求1至6中任一项的化合物的荧光标记物、自旋标记物、重金属标记物或同位素标记物。
PCT/CN2017/116914 2016-12-20 2017-12-18 具有吲哚胺2,3-双加氧酶抑制活性的稠合咪唑化合物 WO2018113624A1 (zh)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP17882589.9A EP3560928B1 (en) 2016-12-20 2017-12-18 Fused imidazole compound having indoleamine 2,3-dioxygenase inhibitory activity
US16/472,185 US10604529B2 (en) 2016-12-20 2017-12-18 Fused imidazole compound having indoleamine 2,3-dioxygenase inhibitory activity

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201611186194.4 2016-12-20
CN201611186194 2016-12-20

Publications (1)

Publication Number Publication Date
WO2018113624A1 true WO2018113624A1 (zh) 2018-06-28

Family

ID=62604862

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/116914 WO2018113624A1 (zh) 2016-12-20 2017-12-18 具有吲哚胺2,3-双加氧酶抑制活性的稠合咪唑化合物

Country Status (5)

Country Link
US (1) US10604529B2 (zh)
EP (1) EP3560928B1 (zh)
CN (1) CN108203438B (zh)
TW (1) TWI671302B (zh)
WO (1) WO2018113624A1 (zh)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3636644A1 (de) * 2018-10-11 2020-04-15 Bayer Aktiengesellschaft Mesoionische imidazopyridine als insektizide
JP2022508532A (ja) * 2018-09-27 2022-01-19 シェンチェン チップスクリーン バイオサイエンシズ カンパニー、リミテッド インドールアミン－２，３－ジオキシゲナーゼ阻害活性を有するキノロン誘導体

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11839659B2 (en) 2020-07-02 2023-12-12 Northwestern University Proteolysis-targeting chimeric molecules (PROTACs) that induce degradation of indoleamine 2,3-dioxygenase (IDO) protein
CN113816965B (zh) * 2021-11-23 2022-03-08 北京鑫开元医药科技有限公司 一种具有ido抑制活性的化合物及其制备方法

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1795187A (zh) 2003-03-27 2006-06-28 兰肯瑙医学研究所 新型ido抑制剂及其使用方法
CN101932325A (zh) 2007-11-30 2010-12-29 新联基因公司 Ido抑制剂
CN102579452A (zh) 2012-01-20 2012-07-18 辽宁思百得医药科技有限公司 色胺酮类化合物的制备方法及其在制备ido抑制剂中的新用途
CN103054870A (zh) 2013-01-08 2013-04-24 复旦大学 色胺酮类化合物作为吲哚胺2,3-双加氧酶抑制剂的用途
CN103547579A (zh) * 2011-04-15 2014-01-29 新联基因公司 用作ido抑制剂的稠合咪唑衍生物
CN105189466A (zh) * 2013-03-14 2015-12-23 新联基因公司 作为色氨酸代谢介导的免疫抑制的抑制剂的三环化合物
WO2016037026A1 (en) * 2014-09-05 2016-03-10 Merck Patent Gmbh Cyclohexyl-ethyl substituted diaza- and triaza-tricyclic compounds as indole-amine-2,3-dioxygenase (ido) antagonists for the treatment of cancer

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10308647B2 (en) * 2015-10-29 2019-06-04 Scifluor Life Sciences, Inc. Fused imidazole derivatives as IDO/TDO inhibitors

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1795187A (zh) 2003-03-27 2006-06-28 兰肯瑙医学研究所 新型ido抑制剂及其使用方法
CN101932325A (zh) 2007-11-30 2010-12-29 新联基因公司 Ido抑制剂
CN103547579A (zh) * 2011-04-15 2014-01-29 新联基因公司 用作ido抑制剂的稠合咪唑衍生物
CN102579452A (zh) 2012-01-20 2012-07-18 辽宁思百得医药科技有限公司 色胺酮类化合物的制备方法及其在制备ido抑制剂中的新用途
CN103054870A (zh) 2013-01-08 2013-04-24 复旦大学 色胺酮类化合物作为吲哚胺2,3-双加氧酶抑制剂的用途
CN105189466A (zh) * 2013-03-14 2015-12-23 新联基因公司 作为色氨酸代谢介导的免疫抑制的抑制剂的三环化合物
WO2016037026A1 (en) * 2014-09-05 2016-03-10 Merck Patent Gmbh Cyclohexyl-ethyl substituted diaza- and triaza-tricyclic compounds as indole-amine-2,3-dioxygenase (ido) antagonists for the treatment of cancer

Non-Patent Citations (13)

* Cited by examiner, † Cited by third party
Title
FRIBERG M. ET AL., INT J CANCER, vol. 101, no. 2, 2002, pages 151 - 155
FUSAO HIRATA ET AL., J BIOL CHEM, vol. 252, no. 13, 1977, pages 4637 - 4642
KING N. J. ET AL., THE INT J BIOCHEM CELL BIOL, vol. 39, no. 12, 2007, pages 2167 - 2172
LEKLEM J. E., AM J CLIN NUTR, vol. 24, no. 6, 1971, pages 659 - 672
MACKENZIE, C. R. ET AL., CURRENT DRUG METABOLISM, vol. 8, 2007, pages 237 - 244
MUNN D. H. ET AL., J EXP MED, vol. 189, no. 9, 1999, pages 1363 - 1372
MUNN D. H. ET AL., SCIENCE, vol. 281, no. 5380, 1998, pages 1191 - 1193
ROY E. J. ET AL., NEUROSCI LETT, vol. 387, no. 2, 2005, pages 95 - 99
See also references of EP3560928A4
TAKIKAWA ET AL., ADV. EXP. MED. BIOL., vol. 467, 1999, pages 241 - 245
TAKIKAWA O. ET AL., EXP. EYE RES., vol. 72, 2001, pages 271 - 277
TEMESS P. ET AL., BLOOD, vol. 105, no. 6, 2005, pages 2480 - 2486
YAMAOTO S. ET AL., J BIOL CHEM, vol. 242, no. 22, 1967, pages 5260 - 5266

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2022508532A (ja) * 2018-09-27 2022-01-19 シェンチェン チップスクリーン バイオサイエンシズ カンパニー、リミテッド インドールアミン－２，３－ジオキシゲナーゼ阻害活性を有するキノロン誘導体
EP3858813A4 (en) * 2018-09-27 2022-06-22 Shenzhen Chipscreen Biosciences Co., Ltd. QUINOLONE DERIVATIVE WITH INDOLEAMINE-2,3-DIOXYGENASE INHIBITED ACTIVITY
JP7455133B2 (ja) 2018-09-27 2024-03-25 シェンチェン チップスクリーン バイオサイエンシズ カンパニー、リミテッド インドールアミン－２，３－ジオキシゲナーゼ阻害活性を有するキノロン誘導体
EP3636644A1 (de) * 2018-10-11 2020-04-15 Bayer Aktiengesellschaft Mesoionische imidazopyridine als insektizide

Also Published As

Publication number Publication date
US20190352307A1 (en) 2019-11-21
EP3560928A4 (en) 2020-09-16
CN108203438A (zh) 2018-06-26
US10604529B2 (en) 2020-03-31
EP3560928A1 (en) 2019-10-30
TWI671302B (zh) 2019-09-11
EP3560928B1 (en) 2021-10-20
TW201927786A (zh) 2019-07-16
CN108203438B (zh) 2021-09-28

Similar Documents

Publication Publication Date Title
CN109311889B (zh) 激活素受体样激酶抑制剂
CA3177261A1 (en) Benzothiazolyl biaryl compound, and preparation method and use
EP3740206B1 (en) Inhibitors of cyclin-dependent kinase 7 (cdk7)
CN113801114B (zh) 稠合二环杂芳基类衍生物、其制备方法及其在医药上的应用
TWI671302B (zh) 具有吲哚胺２，３-雙加氧酶抑制活性的稠合咪唑化合物
WO2013051639A1 (ja) ピラゾロキノリン誘導体
CN110494433B (zh) 布鲁顿酪氨酸激酶抑制剂
TW201704237A (zh) 適用於治療與ｋｉｔ及ｐｄｆｇｒ相關之病症的組合物
PT2997023T (pt) Derivados de bipirazole como inibidores da jak
WO2020035020A1 (zh) 咪唑并吡啶类衍生物及其制备方法和其在医药上的用途
KR20200089718A (ko) 인돌 화합물 및 이의 용도
CN112771046B (zh) 其他被取代的三唑并喹喔啉衍生物
JP5348705B2 (ja) ２−アリールチアゾール−４−カルボキサミド誘導体、それらの製造および医薬としての使用
WO2023246656A1 (zh) Sos1蛋白降解靶向嵌合体及其组合物、制剂和用途
CN113906020A (zh) 用于治疗皮肤病的缓激肽（bk）b2受体拮抗剂的(r)-3-(氯-5-氟-2-((4-(1h-吡唑-1-基)-2-甲基喹啉-8-基氧基)甲基)苯基）吗啉衍生物及相关的化合物
CN112300173B (zh) 一类含氮多环类化合物、制备方法和用途
WO2019100743A1 (zh) 含有苯并呋喃的parp-1和pi3k双靶点抑制剂
WO2022171118A1 (zh) 一种具有抗肿瘤活性的化合物及其用途
CN115536660A (zh) 苄氨基取代的杂多环化合物及其组合物、制剂和用途
JP2018058863A (ja) キナーゼ阻害活性を有するトリプトリン誘導体及びその使用
CN116194103A (zh) 细胞周期蛋白依赖性激酶7（cdk7）非共价抑制剂
KR20200140262A (ko) Tcr-nck 상호 작용의 억제제로서의 크로멘 유도체
WO2023202706A1 (zh) 硒杂环类化合物的盐型和晶型及其应用
WO2023088493A1 (zh) 一种呋喃并吡啶酮类化合物及其应用
CN109422739B (zh) 氘代的吲哚胺2，3-双加氧酶抑制剂及其应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17882589

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2017882589

Country of ref document: EP

Effective date: 20190722