WO2017035982A1 - 一类邻萘醌化合物、其制备方法和医药用途 - Google Patents

一类邻萘醌化合物、其制备方法和医药用途 Download PDF

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WO2017035982A1
WO2017035982A1 PCT/CN2015/097157 CN2015097157W WO2017035982A1 WO 2017035982 A1 WO2017035982 A1 WO 2017035982A1 CN 2015097157 W CN2015097157 W CN 2015097157W WO 2017035982 A1 WO2017035982 A1 WO 2017035982A1
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张晓进
尤启冬
卞金磊
钱雪
王楠
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中国药科大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A61K31/343—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4184—1,3-Diazoles condensed with carbocyclic rings, e.g. benzimidazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4525—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with oxygen as a ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/52—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings condensed with carbocyclic rings or ring systems
    • C07D263/60—Naphthoxazoles; Hydrogenated naphthoxazoles
    • 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
    • C07D307/92—Naphthofurans; Hydrogenated naphthofurans

Definitions

  • the invention relates to the field of medicinal chemistry, in particular to a class of o-naphthoquinone compounds having antitumor activity, including preparation methods of these compounds, which can be activated at a molecular level by a reduced coenzyme I: quinone oxidoreductase (NQO1), in cells Levels and animal levels show a better anti-tumor effect and can be used to prepare anti-tumor drugs.
  • NQO1 quinone oxidoreductase
  • the main need to consider is to selectively kill tumor cells without causing damage to normal tissues.
  • One of the methods is to selectively inhibit tumors based on the bioreduction-activated pathway and utilize the characteristics of reductase with high expression in tumor cells.
  • Workman and Walton first proposed the concept of a bioreductase-based targeted drug in 1989, targeting bioreductases that are highly expressed in tumor cells, and selectively killing the corresponding tumor cells by reducing the substrate. .
  • the quinone oxidoreductase (NQO1) is a reductive metabolic enzyme that is widely used as a selective antitumor. Many solid tumors contain high expression of NQO1, and the NQO1 enzyme itself has the characteristics of biologically activating many anti-tumor agents. Therefore, the anti-tumor effect based on the biological two-electron reduction metabolic enzyme NQO1 has become a hot spot of recent research.
  • orthologous compounds are cytotoxic and can be used as anti-tumor drugs.
  • orthologous compounds such as tanshinone IIA, cryptotanshinone or ⁇ -lapach have shown some inhibitory activity against tumor cell A549.
  • this type of orthoquinone compound can be used as a substrate for the NQO1 enzyme to selectively kill tumor cells by the production of reactive oxygen species mediated by the NQO1 enzyme.
  • This kind of orthoquinone compound can be rapidly and massively reduced to o-diphenol product in tumor cells with high expression of NQO1. The latter undergoes auto-oxidation under the action of cellular oxygen molecules to transform into a ruthenium substrate to form a redox cycle.
  • the invention constructs a skeleton of dimethylnaphthalene orthoquinone compound for the first time on the basis of the structure of tanshinone IIA, and on the basis of retaining the key orthoquinone skeleton of the anti-tumor effect of the natural product tanshinone IIA, and introduces a series of side chains to synthesize a Tanshinone IIA derivative.
  • the compound of the present invention has an antitumor activity comparable to that of tanshinone IIA, and is superior in selectivity to tanshinone IIA, and is expected to be developed into an antitumor drug.
  • the o-naphthoquinone compound of the present invention has a structure represented by Formula I:
  • X represents a carbon atom or a nitrogen atom; when X is a nitrogen atom, R 1 is unsubstituted; when X is a carbon atom, R 1 is hydrogen or a C 1 -C 4 alkyl group;
  • L represents -(CH 2 ) n -, -CO(CH 2 ) n - or -SO 2 (CH 2 ) n -, wherein n represents 0 to 2;
  • R 2 represents hydrogen, methyl, C 1 -C 4 alkoxy, -NR a R b , m represents 0 or 1;
  • Ar represents a benzene ring and or a 5-6 membered aromatic heterocyclic ring;
  • R 4 represents hydrogen, halogen, cyano, nitro, methoxy, C 1 -C 4 alkyl, -NR a R b Or -CH 2 -NR a R b ;
  • R b in R a, R b each independently represent hydrogen, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 hydroxyalkyl group or R a, R b connected to form a 5 to 6-membered nitrogen-containing heterocycle .
  • R 1 preferably represents a methyl group.
  • L preferably represents a bond, -CH 2 -, -CO-, -COCH 2 - or -SO 2 -.
  • R 2 preferably represents hydrogen, -NR a R b , Wherein R a , R b , m, R 4 are as defined in claim 1.
  • R a and R b each preferably independently represent hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl; or preferably R a and R b are bonded to form tetrahydrogen.
  • the compound III is reacted with 1,4-butadiene to obtain a compound IV.
  • the reaction temperature is preferably 80 to 150 ° C
  • the reaction time is preferably 12 to 24 hours
  • the reaction solvent is preferably toluene, benzene, DMF, xylene or the like. 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone should also be added to the reaction.
  • the compound IV is reacted with a dihaloalkane, a halogenated acid chloride or a halosulfonyl chloride to form V.
  • the reaction temperature is 40 to 60 ° C
  • the reaction time is preferably 4 to 12 hours
  • the reaction solvent is ethyl acetate, DMF, dichloromethane, acetone or the like.
  • An inorganic base or an organic base such as sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, triethylamine or the like should also be added to the reaction.
  • Compound V is reacted with a substituted or unsubstituted phenol, a fatty amine or an aromatic amine or other affinity reagent to obtain a target I, a reaction temperature of 20 to 60 ° C, a reaction time of 2 to 8 hours, and a reaction solvent of DMF and acetonitrile. , acetone, tetrahydrofuran, dichloromethane, and the like.
  • An inorganic base or an organic base such as sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, triethylamine or the like should also be added to the reaction.
  • the compound of formula I can be purified by conventional separation methods such as recrystallization, column chromatography and the like.
  • the compound of the present invention may be added into a pharmaceutically acceptable carrier to prepare a common pharmaceutical preparation, such as a tablet, a capsule, a powder, a syrup, a liquid, a suspension, an injection, and may be added to a fragrance, a sweetener, a liquid or Common pharmaceutical excipients such as solid fillers or thinners.
  • a pharmaceutically acceptable carrier such as a tablet, a capsule, a powder, a syrup, a liquid, a suspension, an injection, and may be added to a fragrance, a sweetener, a liquid or Common pharmaceutical excipients such as solid fillers or thinners.
  • the invention has the advantages that: the tanshinone IIA isolated from the natural product can be modified as a raw material, and can be directly synthesized; the structure is novel; the antitumor activity is better than or equivalent to tanshinone IIA; and the compound is double electron redox a substrate for the enzyme NQO1, which is better than the tanshinone IIA by the NQO1 enzyme; the compound of the present invention is a medicine It is obviously superior to tanshinone IIA and is expected to be developed into an anti-tumor drug.
  • the compound of the present invention is used in a clinical dose of 0.01 mg to 1000 mg/day, and may be deviated from this range depending on the severity of the condition or the dosage form.
  • NADPH is oxidized to NADP + in the presence of the enzyme NQO1 (EC 1.6.5.5, purchased by Sigma) and a ruthenium compound, which results in a change in absorbance at 340 nm (monitored by Varioskan Flash) .
  • the compound was first incubated with the enzyme at 37 ° C, the reaction was started after NADPH was added, and the change in luminescence 330 nm was dynamically detected within 3 minutes. The reduction rate is recorded as ⁇ mol NADPH/min/ ⁇ mol NQO1.
  • MTT staining (Cancer Research 47 (1987) 936-942) was used, and the culture time was 72 hours. Inoculates in a 96-well plate at 3.0 ⁇ 10 3 /well, with six gradients for each compound and three replicates for each concentration. The optical density value (OD) was measured by a microplate reader at a wavelength of 560 nm. The tumor cells treated with the solvent control were used as a control group, and the IC 50 was calculated using Grappad Prism 6 software.
  • the compound of the present invention has a strong anti-tumor cell proliferation activity, and its activity is superior to or comparable to that of tanshinone IIA.
  • the ability of the compound to produce reactive oxygen species is assessed at the cellular level (intravenously).
  • the DCFH-DA kit produced by sigma was used to detect whether the compound can increase the ROS level in the tumor cells by using the NQO1 high-expressing lung cancer tumor cells, thereby destroying the redox balance of the tumor cells and achieving the anti-tumor effect.
  • the DCFH-DA kit is a kit for detecting reactive oxygen species using the fluorescent probe DCFH-DA.
  • DCFH-DA itself has no fluorescence and can pass through the cell membrane freely. After entering the cell, it can be hydrolyzed by intracellular esterase to form DCFH. DCFH does not penetrate the cell membrane, making it easy for the probe to be loaded into the cell.
  • Reactive oxygen species in the cells can oxidize non-fluorescent DCFH to produce fluorescent DCF.
  • the level of reactive oxygen species in the cells can be known by detecting the fluorescence of the DCF.
  • Figure 1 shows representative compounds against A549 tumor cells. Multiples of increased sex (relative to blank DMSO).
  • the representative compound of the present invention CPUYB030
  • Figure 1 is a multiple of the increase in activity of representative compounds against A549 tumor cells.
  • Morpholine (26 mL, 0.294 mol) and anhydrous potassium carbonate (2.0 g, 14.7 mmol) were placed in a 250 mL three-necked flask, and a stirrer was added. Stir under nitrogen protection at 0 ° C in an ice bath. When the temperature of the solution dropped to 0 ° C, propionaldehyde (9 mL, 0.125 mol) was slowly injected using a syringe. Be sure to add slowly when adding propionaldehyde. Always pay attention to the change of temperature. Propionaldehyde generally needs to be added within about 1 hour. After the addition was completed, the reaction was further stirred under an ice bath for 2 hours.
  • reaction mixture was filtered, and the solid was filtered and washed with diethyl ether.
  • the filtrate was subjected to rotary evaporation to remove a substance having a low boiling point, and then the residue was subjected to distillation under reduced pressure of bp 88-90 ° C / 25 mmHg to obtain a colorless oily liquid (7.0 g, 45%).
  • Phenylhydrazine (5.0 g, 46.3 mmol) was dissolved in 150 mL of anhydrous dichloromethane and cooled to -5 °C.
  • the distilled 3a (5.9 g, 46.3 mmol) was dissolved in anhydrous dichloromethane (50 mL) and then cooled to -5 ° C.
  • the cooled (E)-4-(prop-1-enyl)morpholine solution was added dropwise to the likewise cooled p-benzoquinone solution with a syringe under nitrogen. During the addition, the solution gradually turns into wine red, which is normal.
  • the burgundy solution was stirred at 0 ° C for 3 hours in an ice bath and then stirred at room temperature overnight. After completion of the reaction, the solid product was filtered, washed with methylene chloride (2.times.10 mL). This crude product was used in the next step without further purification.
  • the morpholine (0.46 mmol), potassium iodide (10 mg, 0.06 mmol) and potassium carbonate (50 mg, 0.38 mmol) were dissolved in N,N-dimethylformamide (5 mL), and 8a (100 mg, 0.38) was added to the reaction mixture. Mm).
  • the reaction solution was reacted at 55 ° C for 1 h. After cooling, the reaction was poured into ice water, extracted with EtOAc (EtOAc)EtOAc.
  • the compound CPUYB002 (38 mg, 30%) was synthesized in the same manner as the compound CPUYB001 by using piperidine (0.46 mmol) instead of morpholine. Mp166-167°C, 1 H NMR (300MHz, CDCl 3 ) ⁇ : 7.83 (s, 1H), 7.52 (s, 1H), 3.62 (s, 2H), 2.54 (s, 4H), 2.37 (s, 3H) , 2.33(s,3H), 2.29(s,3H), 1.67(s,4H), 1.48(s,2H).ESI-HRMS m/z[M+H] + calculated for C 20 H 24 NO 3 : 338.1751, found: 338.1760.
  • the compound CPUYB005 (39 mg, 32%) was synthesized in the same manner as the compound CPUYB001 using pyridine (0.46 mmol). Mp150-151°C, 1 H NMR (300MHz, CDCl 3 ) ⁇ : 7.82 (s, 1H), 7.52 (s, 1H), 3.69 (s, 2H), 2.65 (s, 4H), 2.36 (s, 3H) , 2.32 (s, 3H), 2.28 (s, 3H), 1.85 (s, 4H).
  • the compound CPUYB006 (41 mg, 28%) was synthesized in the same manner as the compound of the compound of the compound of the compound of the compound. Mp 151-152 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.83 (s, 1H), 7.50 (s, 1H), 4.23 (s, 2H), 3.63 (s, 2H), 2.68 (s, 8H) , 2.38(s,3H), 2.33(s,3H), 2.28(s,2H),2.05(s,3H).ESI-HRMS m/z[M+H] + C 22 H 27 N 2 O 4 : 383.1965, found: 383.1963.
  • the compound CPUYB007 (31 mg, 28%) was synthesized in the same manner as the compound CPUYB001 by using methylamine (0.46 mmol) instead of morpholine. Mp 156-157 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.83 (s, 1H), 7.53 (s, 1H), 3.59 (s, 2H), 2.39 (s, 3H), 2.37 (s, 3H) , 2.33 (s, 3H), 2.30 (s, 3H).
  • the compound CPUYB008 (35 mg, 28%) was synthesized in the same manner as the compound (c. Mp 161-162 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.84 (s, 1H), 7.50 (s, 1H), 3.74 (s, 2H), 2.69 (s, 4H), 2.8 (s, 3H) , 2.34 (s, 3H), 2.31 (s, 3H), 1.19 (s, 6H). (m/z) EI-MS: 325 [M] + .
  • the compound CPUYB010 (28 mg, 18%) was synthesized in the same manner as the compound CPUYB001 using o-methoxyaniline (0.46 mmol). Black red solid. Mp 190-191 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.80 (s, 1H), 7.45 (s, 1H), 6.81-6.90 (m, 4H), 4.40 (s, 2H), 3.86 (s, 3H), 2.35 (s, 3H), 2.31 (s, 3H), 2.25 (s, 3H).
  • the compound CPUYB011 (25 mg, 18%) was synthesized in the same manner as the compound CPUYB001 by using fluoroaniline (0.46 mmol) instead of morpholine. Red solid. Mp 180-181 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.82 (s, 1H), 7.46 (s, 1H), 7.19-7.21 (m, 1H), 6.61-6.65 (m, 3H), 4.37 ( s, 2H), 2.38 (s, 3H), 2.33 (s, 3H), 2.28 (s, 3H).
  • the compound CPUY012 (20 mg, 14%) was synthesized in the same manner as the compound CPUYB001 using 2-methyl-4-fluoroaniline (0.46 mmol). Red solid. Mp 188-189°C, 1 H NMR (300MHz, CDCl 3 ) ⁇ : 7.81 (s, 1H), 7.41 (s, 1H), 6.75-6.88 (m, 3H), 4.36 (s, 2H), 2.35 (s) , 3H), 2.31 (s, 3H), 2.26 (s, 3H), 2.21 (s, 3H), m/z (EI-MS): 377 [M] + .
  • the compound CPUYB015 (61 mg, 32%) was synthesized in the same manner as the compound of the compound of the compound of the compound of the compound of the compound. Red solid. Mp 193-194 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.79 (s, 1H), 7.39 (s, 1H), 7.29-7.23 (m, 5H), 3.68 (s, 4H), 3.52 (s, 2H), 2.49 (s, 4H), 2.37 (s, 3H), 2.31 (s, 3H), 2.27 (s, 3H). m/z (EI-MS): 414 [M] + .
  • the compound CPUYB016 (55 mg, 28%) was synthesized in the same manner as the compound of the compound of the compound of the compound of the compound of the compound. Red solid. Mp 190-191 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.79 (s, 1H), 7.39 (s, 1H), 7.29-7.23 (m, 5H), 4.36 (s, 2H), 3.68 (s, 4H), 3.52 (s, 2H), 2.49 (s, 4H), 2.37 (s, 3H), 2.31 (s, 3H), 2.27 (s, 3H), m/z (EI-MS): 428 [M ] + .
  • the compound CPUYB017 (40 mg, 20%) was synthesized by the same method as the compound CPUYB001 using fluorophenylpiperazine (0.46 mmol) instead of morpholine. Red solid. Mp 196-198 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.82 (s, 1H), 7.51 (s, 1H), 6.99-6.85 (m, 4H), 3.67 (s, 2H), 3.17 (s, 4H), 2.73 (s, 4H), 2.36 (s, 3H), 2.32 (s, 3H), 2.30 (s, 3H).m/z (EI-MS): 432 [M] + .
  • valeraldehyde (9.5 mL, 0.125 mol) was used to synthesize a colorless oily liquid 3c (9.43 g, 49%) in the same manner as Compound 3a.
  • the compound 8g was synthesized in the same manner as the compound 8e by using chloropropionyl chloride instead of chloroformyl chloride.
  • the compound CPUYB040 (62 mg, 38%) was synthesized in the same manner as the compound of the compound of the compound of the compound of the compound of the product of the compound of the compound of the compound of the compound of the compound of the compound of Red solid. Mp 190-191 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.82 (s, 1H), 7.47 (s, 1H), 6.69 (s, 2H), 6.42 (s, 2H), 3.66 (s, 2H) , 3.64 (m, 2H), 3.44 (s, 4H), 2.36 (s, 4H), 2.38 (s, 3H), 2.34 (s, 3H), 2.32 (s, 3H), 2.26 (s, 3H). m/z (EI-MS): 521 [M] + .
  • Compound 8j was synthesized in the same manner as Compound 8g by using chloroethanesulfonic acid as the chlorosulfonic acid.
  • the compound CPUYB041 (67 mg, 41%) was synthesized by the same method as the compound CPUYB038, using 4-(dihydroxyethylamino-1-methyl)aniline (0.46 mmol) instead of morpholine. Red solid.
  • the compound CPUYB043 (38 mg, 31%) was synthesized in the same manner as the compound of the compound of the compound of the compound of the compound of the compound of the compound of the compound.
  • the compound CPUYB045 (65 mg, 41%) was synthesized in the same manners as the compound of the compound of the compound of the compound of the compound of the compound of the compound of the compound. Black red solid. Mp 168-169 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.82 (s, 1H), 7.41 (s, 1H), 7.44 - 7.28 (m, 5H), 3.88 (s, 2H), 3.61 (s, 2H), 2.75-2.51 (m, 8H), 2.30 (s, 3H), 2.29 (s, 3H).m/z (EI-MS): 415 [M] + .
  • the compound CPUYB046 (27 mg, 34%) was synthesized by the same method as the compound of the compound of the compound of the compound of the compound of the compound. Black red solid. Mp 172-174 ° C, 1 H NMR (300 MHz, CDCl 3 ) ⁇ : 7.80 (s, 1H), 7.40 (s, 1H), 7.33 - 7.23 (m, 4H), 5.38 (s, 2H), 2.30 (s, 3H), 2.29 (s, 3H).m/z (EI-MS): 351 [M] + .

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Abstract

本发明涉及药物化学领域，具体涉及一类具有抗肿瘤活性的邻萘醌化合物，包括这些化合物制备方法，这些化合物在分子水平可以被还原型辅酶I：醌氧化还原酶(NQO1)活化，在细胞水平和动物水平均表现出较良的抗肿瘤作用，可用于制备抗肿瘤药物。

Description

一类邻萘醌化合物、其制备方法和医药用途 技术领域
本发明涉及药物化学领域，具体涉及一类具有抗肿瘤活性的邻萘醌化合物，包括这些化合物制备方法，这些化合物在分子水平可以被还原型辅酶I：醌氧化还原酶(NQO1)活化，在细胞水平和动物水平均表现出较良的抗肿瘤作用，可用于制备抗肿瘤药物。
背景技术
目前人们在利用小分子来抗肿瘤的策略中，主要需要考虑的是在选择性地杀死肿瘤细胞的同时，而不会对正常组织带来损伤。为了提高药物选择性，我们在设计药物的时候更需要关注肿瘤细胞与正常细胞在生化和生理过程的区别。其中之一的方法就是基于生物还原激活的途径，利用肿瘤细胞中具有高表达的还原酶这一特征，而达到选择性抑制肿瘤的作用。Workman和Walton于1989年首次提出了基于生物还原酶的靶向药物的概念，以在肿瘤细胞中高表达的生物还原酶为靶点，通过对底物进行还原从而选择性地杀死相应的肿瘤细胞。而醌氧化还原酶(NQO1)就是其中被广泛作为选择性抗肿瘤的还原代谢酶。很多实体瘤中都含有高表达的NQO1，并且NQO1酶自身具有可以生物激活许多醌抗肿瘤试剂的特点。因此，基于生物双电子还原代谢酶NQO1的抗肿瘤作用成为最近研究的热点。
而很多邻醌化合物都具有细胞毒性，可作为抗肿瘤药物。例如之前报道的丹参酮IIA、隐丹参酮或β-拉帕醌等邻醌类化合物都对肿瘤细胞A549显示出了一定抑制活性。而最近有文献报道该类型的邻醌类化合物可作为NQO1酶的底物，在NQO1酶的介导下通过产生活性氧的方法，选择性的杀死肿瘤细胞。该类邻醌类化合物能够在NQO1高表达的肿瘤细胞中快速、大量的被还原代谢为邻二酚产物，后者在细胞氧分子作用下发生自氧化重新转变为醌底物，形成氧化还原循环，而这一氧化还原循环过程将产生大量的活性氧。高浓度的活性氧将严重破坏肿瘤细胞的氧化还原微环境平衡，产生氧化应激，进而通过多种途径如降低线粒体的膜电位、激活Caspase-3凋亡通路或直接破坏DNA功能等诱导肿瘤细胞凋亡，发挥选择性抗肿瘤作用。
Figure PCTCN2015097157-appb-000001
发明内容
本发明通过对丹参酮IIA的结构剖析，在保留天然产物丹参酮IIA分子抗肿瘤作用的关键邻醌骨架基础上，首次构建了二甲基萘邻醌化合物骨架，并引入一系列侧链，合成了一类丹参酮IIA衍生物。本发明的化合物具有与丹参酮IIA相当的抗肿瘤活性，并且在选择性上明显优于丹参酮IIA，有望开发成抗肿瘤药物。
本发明的邻萘醌化合物具有通式I所示的结构：
Figure PCTCN2015097157-appb-000002
其中X代表碳原子或氮原子；当X为氮原子时，R1无取代；当X为碳原子时，R1为氢、或C1～C4烷基；
L代表-(CH2)n-、-CO(CH2)n-或-SO2(CH2)n-，其中n代表0～2；
R2代表氢、甲基、C1～C4烷氧基、-NRaRb、
Figure PCTCN2015097157-appb-000003
Figure PCTCN2015097157-appb-000004
m代表0或1；Ar代表苯环及或5～6员芳杂环；R4代表氢、卤素、氰基、硝基、甲氧基、C1～C4烷基、-NRaRb或-CH2-NRaRb；
上述-NRaRb中的Ra、Rb各自独立代表氢、C1～C4烷基、C1～C4羟烷基或Ra、Rb连接形成5～6员含氮杂环。
当X为碳原子；R1优选代表甲基。
L优选代表键、-CH2-、-CO-、-COCH2-或-SO2-。
R2优选代表氢、-NRaRb、
Figure PCTCN2015097157-appb-000005
其中Ra、Rb、m、R4定义同权利要求1。
R2代表-NRaRb时，Ra、Rb优选各自独立代表氢、甲基、乙基、丙基、异丙基、正丁基；或者还优选Ra、Rb连接形成四氢吡咯基、哌啶基、吗啡啉基、哌嗪基或N-甲基哌嗪基。
本发明部分化合物的代号及化学结构如下：
Figure PCTCN2015097157-appb-000006
Figure PCTCN2015097157-appb-000007
Figure PCTCN2015097157-appb-000008
Figure PCTCN2015097157-appb-000009
本发明的化合物可以用下列方法制备：
通式I化合物的制备：
Figure PCTCN2015097157-appb-000010
其中X、R1、R2、L的定义同前。
化合物III与1,4-丁二烯反应得到化合物IV，反应温度优选80～150℃，反应时间优选为12～24h，反应溶剂优选甲苯、苯、DMF、二甲苯等。反应中还应加入2,3-二氯-5,6-二氰基-1,4苯醌。
化合物IV与二卤代烷、卤代酰氯或卤代磺酰氯反应生成V，反应温度为40～60℃，反应时间优选为4-12h，反应溶剂为乙酸乙酯、DMF、二氯甲烷、丙酮等。反应中还应加入无机碱或有机碱，如氢氧化钠、氢氧化钾、碳酸钾、碳酸钠、三乙胺等。
化合物V与取代的或无取代的酚、脂肪胺或芳香胺或其他的亲和试剂反应得到目标物I，反应温度为20～60℃，反应时间为2～8h，反应溶剂可选DMF、乙腈、丙酮、四氢呋喃、二氯甲烷等。反应中还应加入无机碱或有机碱，如氢氧化钠、氢氧化钾、碳酸钾、碳酸钠、三乙胺等。
通式I化合物可以采用常见的分离方法进行纯化，如重结晶、柱层析等。
本发明也包括通式I化合物的水合物、立体异构体、溶剂化物和药学上可接受的盐等。它们具有与通式I化合物同样的药理活性。
本发明所述的化合物可以添加药学上可接受的载体制成常见的药用制剂，如片剂、胶囊、粉剂、糖浆、液剂、悬浮剂、针剂，可以加入香料、甜味剂、液体或固体填料或稀释剂等常用药用辅料。
本发明的优点在于：无需采用分离自天然产物的丹参酮IIA作为原料加以修饰，可以直接通过合成得到；结构新颖；其抗肿瘤活性优于丹参酮IIA或与其相当；且该类化合物为双电子氧化还原酶NQO1的底物，其被NQO1酶代谢速率优于丹参酮IIA；本发明化合物的成药 性明显优于丹参酮IIA，有望开发成为抗肿瘤药物。
本发明的化合物临床所用剂量为0.01mg～1000mg/天，也可根据病情的轻重或剂型的不同偏离此范围。
以下是本发明部分化合物的药理学试验及结果：
(1)化合物被NQO1还原代谢速率的测定：
实验方法：利用NADPH在酶NQO1(EC 1.6.5.5,Sigma购买)及醌化合物的存在下，会被氧化为NADP+，这一过程会导致在340nm下的吸光值发生变化(通过Varioskan Flash监控)。首先将化合物与酶在37℃下孵育，当NADPH加进去之后开始反应，3分钟之内动态检测发光330nm的变化。还原速率记为μmol NADPH/min/μmol NQO1.
表1本发明部分化合物对NQO1酶的代谢速率
Figure PCTCN2015097157-appb-000011
(2)化合物对肿瘤细胞的毒性的测定
实验方法：采用MTT染色法(Cancer Research 47(1987)936-942),培养时间为72小时。在96孔板中分别以3.0×103/well接种，每一个化合物设六个梯度，每一浓度设三复孔。用酶标仪，在波长560nm条件下测定光密度值(OD)。以溶剂对照处理的肿瘤细胞为对照组，用Grappad Prism 6软件计算IC50。
表2本发明部分化合物对NQO1高表达的肿瘤细胞A549的抑制作用(IC50:μM)a:
Figure PCTCN2015097157-appb-000012
Figure PCTCN2015097157-appb-000013
a A549：人肺癌细胞
由表2可见，本发明的化合物具有较强的抗肿瘤细胞增殖的活性，其活性优于丹参酮IIA或与其相当。
(3)化合物通过增加活性氧产生的机制达到抗肿瘤作用
实验方法：1、从酶水平检验化合物产生活性氧的水平。采用细胞色素c检测活性氧的生成，细胞色素c在550nm下具有吸收光，被氧化后其吸收光消失。根据细胞色素c量的变化判断化合物产生活性氧的能力。
表3本发明部分化合物在酶水平产生活性氧的速率：
Figure PCTCN2015097157-appb-000014
2、从细胞水平(静脉内)评价化合物产生活性氧的能力。采用sigma生产的DCFH-DA试剂盒，利用NQO1高表达的肺癌肿瘤细胞，检测化合物是否可以提高肿瘤细胞中的ROS水平，从而破坏肿瘤细胞的氧化还原平衡，达到抗肿瘤作用。DCFH-DA试剂盒是一种利用荧光探针DCFH-DA进行活性氧检测的试剂盒。DCFH-DA本身没有荧光，可以自由穿过细胞膜，进入细胞内后，可以被细胞内的酯酶水解生成DCFH。而DCFH不能通透细胞膜，从而使探针很容易被装载到细胞内。细胞内的活性氧可以氧化无荧光的DCFH生成有荧光的DCF。检测DCF的荧光就可以知道细胞内活性氧水平。图1为代表性化合物对A549肿瘤细胞内活 性提高的倍数(相对于空白DMSO)。
由表3和图1可见，本发明的化合物通过产生ROS达到抗肿瘤作用，其产生ROS的能力优于天然产物丹参酮IIA.
(4)体内抗肿瘤活性研究
实验方法：收集生长旺盛期人肺腺癌细胞，在无菌条件下制备成细胞悬液，接种于裸小鼠腋下。裸小鼠移植瘤用游标卡尺测量移植瘤直径，待肿瘤生长至一定大小后将动物分组，每组5只。使用测量瘤径的方法，动态观察被试物抗肿瘤的效应。空白对照给予等剂量羟丙基-β-环糊精；化合物组：尾静脉注射，隔天一次，持续21天。21天后处死荷瘤裸鼠，并分离瘤块称重。所得数据进行统计学处理(t检验)，计算抑瘤率。同时，对最终剥离的瘤块拍照保存图片。
表4本发明代表性化合物的A549移植瘤的生长抑制率
Figure PCTCN2015097157-appb-000015
由表4可见，本发明的代表性化合物CPUYB030对小鼠移植瘤A549的具有很好的抑制作用，与二期临床药物拉帕醌具有相当的抑制活性。
附图说明
图1为代表性化合物对A549肿瘤细胞内活性提高的倍数
具体实施方式
实施例1
通式I化合物合成路线(A系列化合物的合成)：
Figure PCTCN2015097157-appb-000016
(E)-4-(丙-1-烯基)吗啉(3a)的制备
将吗啉(26mL,0.294mol)和无水碳酸钾(2.0g,14.7mmol)置于250mL的三颈瓶中,加入搅拌子。在氮气保护，0℃冰浴下搅拌。当溶液温度降至0℃后，采用注射器慢慢注入丙醛(9mL,0.125mol)。加入丙醛时一定要缓慢，时刻注意温度的变化，丙醛一般要求约1小时内加完。加入完毕后，反应物继续在冰浴下搅拌2小时。反应到规定时间后，将反应液过滤，滤除固体,并用乙醚润洗固体。将滤液进行旋转蒸发除去低沸点的物质,然后将剩余物质进行减压蒸馏b.p.88-90℃/25mmHg，获得无色油状液体(7.0g,45％)。1H NMR(300MHz,DMSO)δ:5.83(d,1H,J＝15Hz),4.38(dq,1H,J＝12Hz,9Hz),3.58-3.50(m,4H),2.85-2.75(m,2H),2.52-2.44(m,2H),1.60-1.53(m,3H)；m/z(EI-MS):127[M]+。
3-甲基-2-吗啉基-5-羟基-2,3-二氢苯并[1,2-b]呋喃(4a)的制备
将对苯醌(5.0g,46.3mmol)用150mL无水二氯甲烷溶解,并置于冰盐浴下冷却至-5℃。事先将蒸馏出来的3a(5.9g,46.3mmol)用无水的二氯甲烷(50mL)溶解，同样置于冰盐浴下冷却至-5℃。在氮气保护下,用注射器取已冷却的(E)-4-(丙-1-烯基)吗啉溶液逐滴加入同样已冷却的对苯醌溶液中。在滴加的过程中,溶液逐渐变为酒红色,这是正常现象。该酒红色的溶液在冰浴下0℃继续搅拌3小时,接着再室温搅拌过夜。反应完成后，过滤出固体产物，用二氯甲烷(2×10mL)洗涤，烘干后得目标产物，为白色絮状固体(4.0g,30％)。此步粗产物不用进一步纯化，可直接用于下步反应。
3-甲基-5-羟基苯并[1,2-b]呋喃(5a)的制备
4a(4.0g,17.0mmol)溶于20mL四氢呋喃中，将稀盐酸(5％,90mL)滴加到反应液中。反应液回流2h，冷却后用乙酸乙酯(3×30mL)萃取，无水硫酸钠干燥，减压除去溶剂后，用二氯甲烷重结晶，最后得白色晶体(2.0g,79.5％)。m.p.145-147℃,1H NMR(300MHz,CDCl3)δ:7.35(s,1H),7.28(d,J＝8.8Hz,1H),6.90(d,J＝2.7Hz,1H),6.77(dd,J＝8.8,2.7Hz,1H), 4.84(s,1H),2.17(s,3H)；m/z(EI-MS):148[M]+。
3-甲基苯并[1,2-b]呋喃-4,5-二酮(6a)的制备
将化合物5a(300mg,2.03mmol)溶于10mL丙酮中，并将此溶液用冰浴冷却至0℃。将新鲜重结晶制备的费米盐1.8g溶于磷酸二氢钾缓冲液(0.07M KH2PO4溶于100mL水中，调PH值至7.0)中，同样将此溶液用冰浴冷却至0℃。取冷却的费米盐溶液逐滴加入到反应液中。此过程要缓慢，控制温度一直维持在0℃。在滴加过程中，溶液慢慢变为暗红色。滴加完毕后，溶液继续在冰浴下搅拌1h。此时溶液中会出现不溶性物质。将此红色固体物质过滤水洗后收集，即为3-甲基苯并[1,2-b]呋喃-4,5-二酮。得到红色固体(220mg,67％)。m.p.170-172℃,1H NMR(300MHz,CDCl3)δ:7.27(s,1H),7.24(d,J＝10.3Hz,1H),6.12(d,J＝10.3Hz,1H),2.24(s,3H)；m/z(EI-MS):162[M]+。
3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮(7a)的制备
将6a(109mg,0.676mmol)溶于20mL甲苯溶液中，加入二氯二氰基苯醌(DDQ,153mg,0.676mmol)及2,3-二甲基-1,3-丁二烯(0.15mL,1.35mmol)，氮气保护下冷凝回流反应40h，反应结束后，通过柱层析(PE:EA＝20:1)分离纯化得红色目标物(30mg,20％)。m.p.180-182℃，1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.45(s,1H),7.23(s,1H),2.38(s,3H),2.33(s,3H),2.29(s,3H)；m/z(EI-MS):m/z(EI-MS):240[M]+。
2-氯甲基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮(8a)的制备
将7a(50mg,0.208mmol)溶于饱和氯化氢的乙酸乙酯溶液中(20mL),于55℃下反应8h，反应结束后，将反应液倒入饱和碳酸氢钠水溶液中，洗去氯化氢，再用乙酸乙酯(20mL×3)萃取，无水硫酸钠干燥，减压除去乙酸乙酯后，得红色固体，无需进一步纯化，直接投下一步反应。m.p.158-160℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.50(s,1H),4.64(s,2H),2.38(s,3H),2.33(s,3H),2.31(s,3H)；m/z(EI-MS):288[M]+。
化合物CPUYB001的制备
将吗啉(0.46mmol),碘化钾(10mg,0.06mmol)以及碳酸钾(50mg,0.38mmol)溶于N,N-二甲基甲酰胺中(5mL)，向反应液中加入8a(100mg,0.38mmol)。反应液于55℃下反应1h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB001(57mg,45％)。m.p.181-182℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.51(s,1H),3.76(s,4H),3.57(s,2H),2.57(s,4H),2.37(s,3H),2.32(s,3H),2.30(s,3H).m/z(EI-MS):339[M]+,ESI-HRMS m/z[M+Na]+calculated for C20H21NO4Na:362.1363, found:362.1372。
实施例2
化合物CPUYB002的制备
用哌啶(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB002(38mg,30％)。m.p.166-167℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.52(s,1H),3.62(s,2H),2.54(s,4H),2.37(s,3H),2.33(s,3H),2.29(s,3H),1.67(s,4H),1.48(s,2H).ESI-HRMS m/z[M+H]+calculated for C20H24NO3:338.1751,found:338.1760。
实施例3
化合物CPUYB003的制备
用4-甲基哌嗪(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB003(52mg,39％)。m.p.152-153℃,1H NMR(300MHz,CDCl3)δ:7.81(s,1H),7.48(s,1H),3.60(s,2H),2.64(s,8H),2.38(s,3H),2.35(s,3H),2.31(s,3H),2.25(s,3H).ESI-HRMS m/z[M+H]+C21H25N2O3:353.1860,found:353.1873。
实施例4
化合物CPUYB004的制备
用4-乙基哌嗪(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB004(49mg,35％)。m.p.155-157℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.50(s,1H),3.62(s,2H),2.66(s,8H),2.49(s,2H),2.37(s,3H),2.32(s,3H),2.27(s,3H),1.16(t,J＝4.5Hz,3H).ESI-HRMS m/z[M+H]+C22H27N2O3:367.2016,found:367.2032。
实施例5
化合物CPUYB005的制备
用吡咯(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB005(39mg,32％)。m.p.150-151℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.52(s,1H),3.69(s,2H),2.65(s,4H),2.36(s,3H),2.32(s,3H),2.28(s,3H),1.85(s,4H).ESI-HRMS m/z[M+H]+C20H22N2O3:324.1594,found:324.1609。
实施例6
化合物CPUYB006的制备
用4-羟乙基哌嗪(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB006(41mg,28％)。m.p.151-152℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.50(s,1H),4.23(s,2H),3.63(s,2H),2.68(s,8H),2.38(s,3H),2.33(s,3H),2.28(s,2H),2.05(s,3H).ESI-HRMS m/z[M+H]+C22H27N2O4:383.1965,found:383.1963。
实施例7
化合物CPUYB007的制备
用甲胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB007(31mg,28％)。m.p.156-157℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.53(s,1H),3.59(s,2H),2.39(s,3H),2.37(s,3H),2.33(s,3H),2.30(s,3H).ESI-HRMS m/z[M+H]+C18H20N O3:298.1438,found:298.1450。
实施例8
化合物CPUYB008的制备
用二乙胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB008(35mg,28％)。m.p.161-162℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.50(s,1H),3.74(s,2H),2.69(s,4H),2.8(s,3H),2.34(s,3H),2.31(s,3H),1.19(s,6H).(m/z)EI-MS:325[M]+。
实施例9
化合物CPUYB009的制备
用对氟苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB009(28mg,20％)。黑红色固体。m.p.184-185℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),6.93(t,J＝9Hz,2H),6.63-6.68(m,2H),4.29(s,2H),2.38(s,3H),2.30(s,3H),2.29(s,3H).ESI-HRMS m/z[M+Na]+C22H18FNO3Na:386.1163,found:386.1171。
实施例10
化合物CPUYB010的制备
用邻甲氧基苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB010(28mg,18％)。黑红色固体。m.p.190-191℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.45(s,1H),6.81-6.90(m,4H),4.40(s,2H),3.86(s,3H),2.35(s,3H),2.31(s,3H),2.25(s,3H).ESI-HRMS m/z[M+H]+C23H21NO4Na:398.1363,found:398.1372。
实施例11
化合物CPUYB011的制备
用间氟苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB011(25mg,18％)。红色固体。m.p.180-181℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),7.19-7.21(m,1H),6.61-6.65(m,3H),4.37(s,2H),2.38(s,3H),2.33(s,3H),2.28(s,3H).ESI-HRMS m/z[M+Na]+C22H18FNO3Na:386.1163,found:386.1185。
实施例12
化合物CPUYB012的制备
用2-甲基-4-氟苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUY012(20mg,14％)。红色固体。m.p 188-189℃,1H NMR(300MHz,CDCl3)δ:7.81(s,1H),7.41(s,1H),6.75-6.88(m,3H),4.36(s,2H),2.35(s,3H),2.31(s,3H),2.26(s,3H),2.21(s,3H).m/z(EI-MS):377[M]+。
实施例13
化合物CPUYB013的制备
用2,4-二氯苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB013(30mg,19％)。红色固体。m.p.190-191℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.43(s,1H),7.31(s,1H),7.17(d,J＝7.2Hz,1H),6.77(d,J＝7.2Hz,1H),4.41(s,2H),2.38(s,3H),2.33(s,6H).m/z(EI-MS):413[M]+。
实施例14
化合物CPUYB014的制备
用4-甲氧基苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB014(21mg,15％)。红色固体。m.p.190-191℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.45(s,1H),6.90-6.81(m,4H),4.40(s,2H),3.86(s,3H),2.35(s,3H),2.31(s,3H),2.25(s,3H).m/z(EI-MS):375[M]+。
实施例15
化合物CPUYB015的制备
用对苯基哌嗪(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物 CPUYB015(61mg,32％)。红色固体。m.p.193-194℃,1H NMR(300MHz,CDCl3)δ:7.79(s,1H),7.39(s,1H),7.29-7.23(m,5H),3.68(s,4H),3.52(s,2H),2.49(s,4H),2.37(s,3H),2.31(s,3H),2.27(s,3H).m/z(EI-MS):414[M]+。
实施例16
化合物CPUYB016的制备
用对苄基哌嗪(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB016(55mg,28％)。红色固体。m.p.190-191℃,1H NMR(300MHz,CDCl3)δ:7.79(s,1H),7.39(s,1H),7.29-7.23(m,5H),4.36(s,2H),3.68(s,4H),3.52(s,2H),2.49(s,4H),2.37(s,3H),2.31(s,3H),2.27(s,3H).m/z(EI-MS):428[M]+。
实施例17
化合物CPUYB017的制备
用对氟苯基哌嗪(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB017(40mg,20％)。红色固体。m.p.196-198℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.51(s,1H),6.99-6.85(m,4H),3.67(s,2H),3.17(s,4H),2.73(s,4H),2.36(s,3H),2.32(s,3H),2.30(s,3H).m/z(EI-MS):432[M]+。
实施例18
化合物CPUYB018的制备
用3-氯-4-氟苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB018(24mg,13％)。红色固体。m.p.196-198℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.42(s,1H),7.01(d,J＝8.8Hz,1H),6.79-6.76(m,1H),6.57-6.55(m,1H),4.30(s,2H),2.37(s,3H),2.33(s,6H).m/z(EI-MS):397[M]+。
实施例19
化合物CPUYB019的制备
用4-硝基苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB019(37mg,19％)。红色固体。m.p.196-198℃,1H NMR(300MHz,CDCl3)δ:7.78(s,1H),7.37(s,1H),7.30(d,J＝8.8Hz,2H),6.60(d,J＝8.8Hz,2H),4.29(s,2H),2.34(s,3H),2.33(d,J＝2.6Hz,6H).m/z(EI-MS):390[M]+。
实施例20
化合物CPUYB020的制备
用2,4-二氟苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB020(40mg,23％)。红色固体。m.p.196-198℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.43(s,1H),6.88-6.70(m,3H),4.34(s,2H),2.37(s,3H),2.32(d,J＝2.7Hz,6H).m/z(EI-MS):381[M]+。
实施例21
化合物CPUYB021的制备
用4-腈基苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB021(37mg,22％)。红色固体。m.p.196-198℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.47(s,1H),7.49(d,J＝6.8Hz,2H),7.37(s,1H),6.70(d,J＝6.8Hz,2H),4.37(d,J＝5.3Hz,2H),2.35(s,3H),2.32(s,6H).m/z(EI-MS):370[M]+。
实施例22
化合物CPUYB022的制备
用3-甲氧基苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB022(41mg,24％)。红色固体。m.p.196-198℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.47(s,1H),6.88(m,4H),4.33(s,2H),3.78(s,3H),2.35(s,3H),2.33(s,3H),2.26(s,3H).m/z(EI-MS):375[M]+。
实施例23
化合物CPUYB023的制备
用3,4-二氯苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB023(38mg,20％)。红色固体。m.p.196-198℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.41(s,1H),7.24(s,1H),6.84(d,J＝2.5Hz,1H),6.57-6.54(m,1H),4.30(s,2H),4.14(brs,1H),2.37(s,3H),2.33(s,6H).m/z(EI-MS):413[M]+。
实施例24
化合物CPUYB024的制备
用4-吗啉甲基-苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物 CPUYB024(35mg,17％)。红色固体。m.p.180-181℃.1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.43(s,1H),7.18(d,J＝8.4Hz,2H),6.69(d,J＝8.4Hz,2H),4.43(s,2H),3.74-3.70(m,4H),3.42(s,2H),2.44-2.39(m,4H),2.36(s,3H),2.32(s,3H),2.28(s,3H).m/z(EI-MS):444[M]+。
实施例25
化合物CPUYB025的制备
用4-对乙基哌嗪基甲基-苯胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB025(65mg,30％)。红色固体。m.p.180-181℃.1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),7.07(d,J＝6.6Hz,2H),6.55(d,J＝6.6Hz,2H),4.38(s,2H),3.18(s,4H),2.48-2.52(m,4H),2.35-2.38(m,7H),2.31(s,3H),2.26(s,3H),2.21(s,3H),1.02(t,J＝7.8Hz,3H).m/z(EI-MS):471[M]+。
实施例26
化合物CPUYB026的制备
用4-氟苄胺(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB026(31mg,18％)。红色固体。m.p.180-181℃.1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),7.35-7.28(m,2H),7.04(t,J＝8.5Hz,2H),3.81(s,4H),2.37(s,3H),2.32(s,3H),2.21(s,3H).m/z(EI-MS):377[M]+。
实施例27
化合物CPUYB027的制备
用4-异丁醇(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB027(35mg,22％)。红色固体。m.p.162-163℃.1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),3.90(m,2H),2.37(s,3H),2.34(s,3H),1.93-1.89(m,1H),0.91(s,6H).m/z(EI-MS):312[M]+。
实施例28
化合物CPUYB028的制备
用4-氟苯酚(0.46mmol)代替吗啉，以与化合物CPUYB001相同的方法合成化合物CPUYB026(25mg,15％)。红色固体。m.p.180-181℃.1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),6.93(t,J＝9Hz,2H),6.63-6.68(m,2H),5.17(s,2H),2.38(s,3H),2.30(s,3H),2.29(s,3H).m/z(EI-MS):364[M]+。
实施例29
(E)-4-(丁-1-烯基)吗啉(3b)的制备
丁醛(9.2mL,0.125mol)代替丙醛，以与化合物3a相同的方法合成无色油状液体3b(8.47g,48％)。1H NMR(300MHz,DMSO)δ:5.83(d,J＝15Hz,1H),4.46-4.44(m,1H),3.65-3.55(m,4H),3.30-3.10(m,4H),2.14-2.00(m,2H),1.60-1.53(t,J＝7.8Hz,3H)；m/z(EI-MS):141[M]+。3-乙基-2-吗啉基-5-羟基-2,3-二氢苯并[1,2-b]呋喃(4b)的制备
用3b(6.5g,46.3mmol)代替3a，以与化合物4a相同的方法合成化合物4b(4.6g,36％)。3-乙基-5-羟基苯并[1,2-b]呋喃(5b)的制备
用4b代替4a外，以与化合物5a相同的方法合成化合物5b。白色晶体(2.3g,78％)。m.p.148-149℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.49(d,J＝8.8Hz,1H),6.80(d,J＝2.7Hz,1H),6.77(dd,J＝8.8,2.7Hz,1H),4.84(s,1H),2.60(q,J＝7.8Hz,2H),1.25(s,3H)；m/z(EI-MS):162[M]+。
3-乙基苯并[1,2-b]呋喃-4,5-二酮(6b)的制备
用5b代替5a外，以与化合物6a相同的方法合成化合物6b。红色固体(230mg,70％)。m.p.174-175℃,1H NMR(300MHz,CDCl3)δ:7.47(s,1H),7.34(d,J＝10.3Hz,1H),6.22(d,J＝10.3Hz,1H),2.60(q,J＝7.8Hz,2H),1.24(s,3H)；m/z(EI-MS):176[M]+。
3-乙基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮(7b)的制备
用6b代替6a外，以与化合物7a相同的方法合成化合物7b。为红色目标物(35mg,21％)。m.p.183-185℃,1H NMR(300MHz,CDCl3)δ:7.86(s,1H),7.54(s,1H),7.41(s,1H),2.38(s,3H),2.33(s,3H),2.29(q,J＝6.9Hz,3H),1.25(t,J＝6.9Hz,3H)；m/z(EI-MS):m/z(EI-MS):254[M]+。
2-氯甲基-3-乙基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮(8b)的制备
用7b代替7a外，以与化合物8a相同的方法合成化合物8b。m.p.156-157℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.54(s,1H),4.49(s,2H),2.62(q,J＝7.8Hz,2H),2.33(s,3H),2.31(s,3H),1.25(t,J＝7.8Hz,3H)；m/z(EI-MS):302[M]+。
化合物CPUYB029的制备
将吗啉(0.46mmol),碘化钾(10mg,0.06mmol)以及碳酸钾(50mg，0.38mmol)溶于N,N- 二甲基甲酰胺中(5mL)，向反应液中加入2-氯甲基-3-乙基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮8b(100mg,0.38mmol)。反应液于55℃下反应1h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB001(57mg,45％)。m.p.181-182℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.51(s,1H),3.76(s,4H),3.57(s,2H),2.60(q,J＝7.8Hz,2H),2.57(s,4H),2.37(s,3H),2.32(s,3H),2.30(t,J＝7.8Hz,3H).m/z(EI-MS):353[M]+,ESI-HRMS m/z[M+Na]+calculated for C21H23NO4Na:376.1519,found:376.1523。
实施例30
化合物CPUYB030的制备
用4-氟苯胺(0.46mmol)代替吗啉，以与化合物CPUYB029相同的方法合成化合物CPUYB030(31mg,18％)。红色固体。m.p.186-187℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),6.93(t,J＝9Hz,2H),6.63-6.68(m,2H),4.29(s,2H),2.60(q,J＝7.8Hz,2H),2.38(s,3H),2.30(s,3H),2.29(t,J＝7.8Hz,3H).ESI-HRMS m/z[M+Na]+C23H20FNO3Na:400.1319,found:400.1322.
实施例31
(E)-4-(戊-1-烯基)吗啉(3c)的制备
用戊醛(9.5mL,0.125mol)代替丙醛，以与化合物3a相同的方法合成无色油状液体3c(9.43g,49％)。1H NMR(300MHz,DMSO)δ:5.84(d,J＝15Hz,1H),4.55-4.44(m,1H),3.65-3.50(m,4H),3.35-3.30(m,4H),2.14-2.00(m,2H),1,46-1.40(m,2H),0.90(t,J＝7.8Hz,3H)；m/z(EI-MS):155[M]+。
3-丙基-2-吗啉基-5-羟基-2,3-二氢苯并[1,2-b]呋喃(4c)的制备
用3c(7.2g,46.3mmol)代替3a，以与化合物4a相同的方法合成化合物4c(4.9g,40％)。
3-丙基-5-羟基苯并[1,2-b]呋喃(5c)的制备
用4c代替4a，以与化合物5a相同的方法合成化合物5c。白色絮状物(1.20g,40％)。m.p.149-151℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.49(d,J＝8.8Hz,1H),6.80(d,J＝2.7Hz,1H),6.77(dd,J＝8.8,2.7Hz,1H),5.35(s,1H),2.62(q,J＝7.8Hz,2H),1.65-1.60(m,2H),1.11-0.90(M,3H)；m/z(EI-MS):176[M]+。
3-丙基苯并[1,2-b]呋喃-4,5-二酮(6c)的制备
用5c代替5a，以与化合物6a相同的方法合成化合物6c。红色固体(174mg,45％)。m.p.177-178℃,1H NMR(300MHz,CDCl3)δ:7.46(s,1H),7.37(d,J＝10.3Hz,1H),6.24(d,J＝10.3Hz,1H),2.62(q,J＝7.8Hz,2H),1.65-1.60(m,2H),0.90-0.85(m,3H)；m/z(EI-MS):190[M]+。
3-丙基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮(7c)的制备
用6c代替6a，以与化合物7a相同的方法合成化合物7c。为红色目标物(45mg,25％)。m.p.184-186℃,1H NMR(300MHz,CDCl3)δ:7.86(s,1H),7.54(s,1H),7.41(s,1H),2.38(s,3H),2.33(s,3H),2.62(t,J＝6.9Hz,3H),1.65-1.60(m,2H),0.90(t,J＝6.9Hz,3H)；m/z(EI-MS):m/z(EI-MS):268[M]+。
2-氯甲基-3-丙基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮(8c)的制备
用7c代替7a，以与化合物8a相同的方法合成化合物8c。m.p.159-160℃,1H NMR(300MHz,CDCl3)δ:7.86(s,1H),7.54(s,1H),4.49(s,2H),2.62(q,J＝7.8Hz,2H),2.33(s,3H),2.31(s,3H),1.65-1.61(m,2H),0.90(t,J＝7.8Hz,3H)；m/z(EI-MS):302[M]+。
化合物CPUYB031的制备
将吗啉(0.46mmol),碘化钾(10mg,0.06mmol)以及碳酸钾(50mg,0.38mmol)溶于DMF中(5mL)，向反应液中加入2-氯甲基-3-丙基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮8c(100mg,0.38mmol)。反应液于55℃下反应1h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB0031(57mg,45％)。m.p.181-182℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.51(s,1H),3.76(s,2H),3.65-3.57(m,4H),2.62(t,J＝7.8Hz,2H),2.57(m,4H),2.37(s,3H),2.32(s,3H),1.65(m,2H),0.90(t,J＝7.8Hz,3H).ESI-HRMS m/z[M+Na]+calculated for C21H23NO4Na:390.1676,found:390.1670。
实施例32
(E)-4-(己-1-烯基)吗啉(3d)的制备
用己醛(9.8mL,0.125mol)代替丙醛，以与化合物3a相同的方法合成无色油状液体3d(6.21g,32％)。1H NMR(300MHz,DMSO)δ:5.84(d,J＝15Hz,1H),4.50-4.44(m,1H),3.65-3.50(m,4H),3.34-3.30(m,4H),2.18-2.10(m,2H),1,36-1.31(m,2H),0.96-0.90(m,3H)；m/z(EI-MS):169[M]+。
3-丁基-2-吗啉基-5-羟基-2,3-二氢苯并[1,2-b]呋喃(4d)的制备
用3d(7.8g,46.3mmol)代替3a，以与化合物4a相同的方法合成化合物4d(4.9g,38％)。3-丁基-5-羟基苯并[1,2-b]呋喃(5d)的制备
用4d代替4a，以与化合物5a相同的方法合成化合物5d。白色絮状物(1.13g,35％)。m.p.154-156℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.49(d,J＝8.8Hz,1H),6.80(d,J＝2.7Hz,1H),6.77(dd,J＝8.8,2.7Hz,1H),5.35(s,1H),2.62(q,J＝7.8Hz,2H),1.65-1.60(m,2H),1.34-1.30(m,2H),1.11-0.90(M,3H)；m/z(EI-MS):190[M]+。
3-丁基苯并[1,2-b]呋喃-4,5-二酮(6d)的制备
用5d代替5a，以与化合物6a相同的方法合成化合物6d。红色固体(157mg,38％)。m.p.155-157℃,1H NMR(300MHz,CDCl3)δ:7.46(s,1H),7.37(d,J＝10.3Hz,1H),6.24(d,J＝10.3Hz,1H),2.62(q,J＝7.8Hz,2H),1.65-1.60(m,2H),1.31-1.27(m,2H),0.90-0.85(m,3H)；m/z(EI-MS):204[M]+。
3-丁基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮(7d)的制备
用6d代替6a，以与化合物7a相同的方法合成化合物7d。为红色目标物(51mg,27％)。m.p.179-180℃,1H NMR(300MHz,CDCl3)δ:7.86(s,1H),7.54(s,1H),7.41(s,1H),2.38(s,3H),2.33(s,3H),2.62(t,J＝6.9Hz,3H),1.65-1.60(m,2H),1.34-1.30(m,2H),0.90(t,J＝6.9Hz,3H)；m/z(EI-MS):m/z(EI-MS):282[M]+。
2-氯甲基-3-丁基-7,8-二甲基萘[1,2-b]呋喃-4,5-二酮(8d)的制备
用7d代替7a，以与化合物8a相同的方法合成化合物8d。m.p.151-152℃,1H NMR(300MHz,CDCl3)δ:7.86(s,1H),7.54(s,1H),4.49(s,2H),2.62(q,J＝7.8Hz,2H),2.33(s,3H),2.31(s,3H),1.65-1.61(m,2H),1.33-1.31(m,2H),0.90(t,J＝7.8Hz,3H)；m/z(EI-MS):330[M]+。
化合物CPUYB032的制备
用4-对甲基哌嗪基甲基-苯胺(0.46mmol)代替吗啉，以与化合物CPUYB031相同的方法合成化合物CPUYB032(28mg,13％)。红色固体。m.p.180-181℃.1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),7.07(d,J＝6.6Hz,2H),6.55(d,J＝6.6Hz,2H),4.38(s,2H),3.18(s,4H),2.62(m,2H),2.48-2.52(m,4H),2.35-2.38(m,7H),2.31(s,3H),2.26(s,3H),2.21(s,3H),1.59(m,2H),1.31(m,2H),1.02(t,J＝7.8Hz,3H).m/z(EI-MS):499[M]+。
实施例33
2-氯甲酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮8e的制备
将上步化合物3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮7a(50mg,0.208mmol)溶于二氯甲烷溶液中(20mL)，冰浴下滴加氯甲酰氯，滴加完毕后室温搅拌1h，反应结束后，将反应液倒入冰水中，再用乙酸乙酯(20mL×3)萃取，无水硫酸钠干燥，减压除去乙酸乙酯后，得红色固体。m.p.150-152℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.50(s,1H),4.64(s,2H),2.38(s,3H),2.33(s,3H),2.51(s,3H)；m/z(EI-MS):302[M]+。
化合物CPUYB033的制备
将4-氟苯胺(0.46mmol),碘化钾(10mg,0.06mmol)以及碳酸钾(50mg，0.38mmol)溶于DMF中(5mL)，向反应液中加入2-氯甲酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮8e(100mg,0.38mmol)。反应液于55℃下反应1h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB0033(57mg,45％)。m.p.172-174℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),6,93(t,J＝9Hz,2H),6.63-6,68(m,2H),2.38(s,3H),2.30(s,3H),2.29(s,3H).m/z(EI-MS):377[M]+。
实施例34
化合物CPUYB034的制备
用4-氟苄胺(0.15mmol)代替4-氟苯胺，以与化合物CPUYB033相同的方法合成化合物CPUYB034(21mg,38％)。红色固体。m.p.190-191℃,1H NMR(300MHz,CDCl3)δ:7.88(s,1H),7.50(s,1H),7.43-7.38(m,2H),7.10(t,J＝8.6Hz,2H),6.71(brs,1H),4.66(d,J＝6.1Hz,2H),2.71(s,3H),2.40(s,3H),2.36(s,3H).m/z(EI-MS):391[M]+。
实施例35
化合物CPUYB035的制备
用4-(二羟乙基胺基-1-甲基)苯胺(0.15mmol)代替4-氟苯胺，以与化合物CPUYB033相同的方法合成化合物CPUYB035(27mg,41％)。红色固体。m.p.179-181℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),7.07(d,J＝6.6Hz,2H),6.55(d,J＝6.6Hz,2H),3.66(s,2H),3.45(t,J＝8.1Hz,4H),2.53(t,J＝8.1Hz,4H),2.31(s,3H),2.26(s,3H),2.21(s,3H).m/z(EI-MS):476[M]+.
实施例36
2-氯乙酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮8f的制备
用氯乙酰氯代替氯甲酰氯外，以与化合物8e相同的方法合成化合物8f。m.p.151-152℃, 1H NMR(300MHz,CDCl3)δ:7.86(s,1H),7.54(s,1H),4.83(s,2H),2.52(s,3H),2.33(s,3H),2.31(s,3H)；m/z(EI-MS):316[M]+。
化合物CPUYB036的制备
将4-氟苯胺(0.46mmol),碘化钾(10mg,0.06mmol)以及碳酸钾(50mg,0.38mmol)溶于DMF中(5mL)，向反应液中加入2-氯乙酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮8f(120mg,0.38mmol)。反应液于55℃下反应1h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB0036(57mg,45％)。m.p.172-174℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),6.93(t,J＝9Hz,2H),6.63-6,68(m,2H),4.74(s,2H),2.38(s,3H),2.30(s,3H),2.29(s,3H).m/z(EI-MS):391[M]+。
实施例37
2-氯丙酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮(8g)的制备
用氯丙酰氯代替氯甲酰氯，以与化合物8e相同的方法合成化合物8g。m.p.161-162℃,1H NMR(300MHz,CDCl3)δ:7.88(s,1H),7.55(s,1H),3.70(t,J＝6.9Hz,2H),3.02(t,J＝6.9Hz,2H),2.51(s,3H),2.34(s,3H)；m/z(EI-MS):330[M]+。
化合物CPUYB037的制备
将4-氟苯胺(0.46mmol),碘化钾(10mg,0.06mmol)以及碳酸钾(50mg,0.38mmol)溶于DMF中(5mL)，向反应液中加入2-氯丙酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮8g(125mg,0.38mmol)。反应液于55℃下反应1h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB0037(69mg,45％)。m.p.172-174℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),6.93(t,J＝9Hz,2H),6.63-6,68(m,2H),4.74(s,2H),3.40-3.36(m,4H),2.38(s,3H),2.30(s,3H),2.29(s,3H).m/z(EI-MS):405[M]+。
实施例38
2-氯磺酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮(8h)的制备
将3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮7a(50mg,0.208mmol)溶于二氯甲烷溶液中(20mL)，冰浴下滴加氯磺酸，滴加完毕后室温搅拌2h，反应结束后，将反应液倒入冰水中，再用乙酸乙酯(20mL×3)萃取，无水硫酸钠干燥，减压除去乙酸乙酯后，得红色固体。m.p.150-152℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.50(s,1H),2.38(s,3H),2.33(s,3H), 1.39(s,3H)；m/z(EI-MS):338[M]+。
化合物CPUYB038的制备
将吗啉(0.46mmol),KI(10mg,0.06mmol)以及碳酸钾(50mg,0.38mmol)溶于DMF中(5mL)，向反应液中加入2-氯甲磺酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮8h(0.38mmol)。反应液于55℃下反应1h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB038(66mg,45％)。m.p.176-177℃.1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),3.51-3.65(m,8H),2.39(s,3H),2.34(s,3H),2.30(s,3H).m/z(EI-MS):389[M]+。
实施例39
化合物CPUYB039的制备
用4-氟苯胺(0.46mmol)代替吗啉，以与化合物CPUYB038相同的方法合成化合物CPUYB039(45mg,29％).m.p.172-174℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),6,92(t,J＝6.99Hz,2H),6.63-6,68(m,2H),2.37(s,3H),2.30(s,3H),2.28(s,3H).m/z(EI-MS):413[M]+。
实施例40
2-氯甲磺酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮(8i)的制备
将3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮7a(50mg,0.208mmol)溶于二氯甲烷溶液中(20mL)，冰浴下滴加氯磺酸，滴加完毕后室温搅拌2h，反应结束后，将反应液倒入冰水中，再用乙酸乙酯(20mL×3)萃取，无水硫酸钠干燥，减压除去乙酸乙酯后，得红色固体。m.p.150-152℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.50(s,1H),2.38(s,3H),2.33(s,3H),1.39(s,3H)；m/z(EI-MS):338[M]+。
化合物CPUYB040的制备
用4-(4-甲基哌嗪-1-甲基)苯胺(0.46mmol)代替吗啉，8i代替8h，以与化合物CPUYB038相同的方法合成化合物CPUYB040(62mg,38％)。红色固体。m.p.190-191℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.47(s,1H),6.69(s,2H),6.42(s,2H),3.66(s,2H),3.64(m,2H),3.44(s,4H),2.36(s,4H),2.38(s,3H),2.34(s,3H),2.32(s,3H),2.26(s,3H).m/z(EI-MS):521[M]+。
实施例41
2-氯乙磺酰基-3,7,8-三甲基萘[1,2-b]呋喃-4,5-二酮8j的制备
用氯乙磺酸替氯磺酸，以与化合物8g相同的方法合成化合物8j。m.p.161-162℃,1H NMR(300MHz,CDCl3)δ:7.88(s,1H),7.55(s,1H),5.12(s,2H),2.36(s,3H),2.34(s,3H),1.93(s,3H)；m/z(EI-MS):352[M]+。
化合物CPUYB041的制备
用4-(二羟乙基胺基-1-甲基)苯胺(0.46mmol)代替吗啉,用8i代替8f，以与化合物CPUYB038相同的方法合成化合物CPUYB041(67mg,41％)。红色固体。m.p.179-181℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.46(s,1H),7.07(d,J＝6.6Hz,2H),6.55(d,J＝6.6Hz,2H),3.64(m,2H),3.62(s,2H),3.45(t,J＝8.1Hz,4H),2.53(t,J＝8.1Hz,4H),2.31(s,3H),2.26(s,3H),2.21(s,3H).m/z(EI-MS):540[M]+。
实施例42
通式I化合物合成路线(B系列化合物的合成)：
Figure PCTCN2015097157-appb-000017
化合物(9)的制备
将化合物1(2.9mmol)溶于THF/H2O中，将用水溶解的叠氮化钠(0.625g,8.7mmol)逐滴加入到反应液中，室温搅拌反应，TLC板监控反应，反应结束后将反应液倒入水中，再用乙酸乙酯萃取，饱和碳酸钠洗，无水硫酸钠干燥，旋干得粗品，用乙醇重结晶最终得橘黄色固体9(380mg,76％).m.p.75-76℃,1H NMR(300MHz,CDCl3)δ:6.83(s,2H),5.52(s,1H).m/z(EI-MS):123[M]+。
化合物(10)的制备
将化合物9(0.58mmol)溶解于醋酸中，将溶有溴化氢(0.058mmol)和氯乙醛(2.9mmol)的醋酸溶液滴加到反应液中，室温搅拌反应，TLC监控反应，反应结束后将反应液倒入水中，用乙酸乙酯萃取，有机层用碳酸氢钠洗，饱和氯化钠洗，无水硫酸钠干燥，旋干，得粗品， 用二氯甲烷重结晶，得粉红色固体13(50mg,47％)。m.p.110-111℃.1H NMR(300MHz,CDCl3)δ:7.62(d,J＝4.4Hz,1H),6.95(d,J＝4.4Hz,1H),6.73(s,1H),5.11(s,2H).m/z(EI-MS):183[M]+。
化合物(11)的制备
将化合物10(0.23mmol)溶于适量丙酮中，溶有费米盐(0.70mmol)的磷酸二氢钾水溶液缓慢滴加到反应液中，室温搅拌，TLC板监控，反应结束后，略微旋去丙酮，有固体析出，将固体抽滤出来得红色固体11(26mg,57.8％)。m.p.140-141℃.1H NMR(300MHz,CDCl3)δ:7.39(d,J＝9.0Hz,1H),6.24(d,J＝9.0Hz,1H),5.09(s,2H).m/z(EI-MS):196[M]+。
化合物(12)的制备
将化合物11(0.676mmol)溶于20mL甲苯溶液中，加入二氯二氰基苯醌(DDQ,0.676mmol)及2,3-二甲基-1,3-丁二烯(0.15mL,1.35mmol)，氮气保护下冷凝回流反应10h，反应结束后，通过柱层析(PE:EA＝20:1)分离纯化得红色目标物12(30mg,20％)。m.p.170-172℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.55(s,1H),5.11(s,2H),2.34(s,3H),2.33(s,3H).m/z(EI-MS):275[M]+。
化合物CPUYB042的制备
将二甲胺(0.46mmol),KI(10mg,0.06mmol)以及碳酸钾(50mg,0.38mmol)溶于DMF(5mL)中，向反应液中加入12(0.38mmol)。反应液于55℃下反应2h。冷却后，将反应也倒入冰水中，用乙酸乙酯萃取，饱和食盐水洗，无水硫酸钠干燥，过滤浓缩后得红色目标物CPUYB042(56mg,45％)。m.p.181-182℃,1H NMR(300MHz,CDCl3)δ:7.83(s,1H),7.53(s,1H),3.59(s,2H),2.45(s,6H),2.33(s,3H),2.30(s,3H),m/z(EI-MS):284[M]+。
实施例43
化合物CPUYB043的制备
用吗啉(0.46mmol)代替二甲胺，以与化合物CPUYB042相同的方法合成化合物CPUYB043(38mg,31％)。m.p.156-157℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.51(s,1H),3.84(s,2H),3.74(s,4H),2.65(s,4H),2.32(s,3H),2.30(s,3H).m/z(EI-MS):326[M]+。
实施例44
化合物CPUYB044的制备
用间氟苯胺(0.46mmol)代替二甲胺，以与化合物CPUYB042相同的方法合成化合物CPUYB044(27mg,20％)。黑红色固体。m.p.182-184℃,1H NMR(300MHz,CDCl3)δ:7.84(s,1H),7.40(s,1H),7.11-7.05(m,1H),6.79(t,J＝6.6Hz,1H),6.52(d,J＝7.8Hz,1H),6.35(t,J＝9.5Hz,1H),4.60(d,J＝6.3Hz,2H).,2.30(s,3H),2.29(s,3H).m/z(EI-MS):350[M]+。
实施例45
化合物CPUYB045的制备
用对苄基哌嗪(0.46mmol)代替二甲胺，以与化合物CPUYB042相同的方法合成化合物CPUYB045(65mg,41％)。黑红色固体。m.p.168-169℃,1H NMR(300MHz,CDCl3)δ:7.82(s,1H),7.41(s,1H),7.44-7.28(m,5H),3.88(s,2H),3.61(s,2H),2.75-2.51(m,8H),2.30(s,3H),2.29(s,3H).m/z(EI-MS):415[M]+。
实施例46
化合物CPUYB046的制备
用对氟苯酚(0.46mmol)代替二甲胺，以与化合物CPUYB042相同的方法合成化合物CPUYB046(27mg,34％)。黑红色固体。m.p.172-174℃,1H NMR(300MHz,CDCl3)δ:7.80(s,1H),7.40(s,1H),7.33-7.23(m,4H),5.38(s,2H),2.30(s,3H),2.29(s,3H).m/z(EI-MS):351[M]+。

Claims (8)

  1. 通式(I)的邻萘醌化合物或其药学上可接受的盐：
    Figure PCTCN2015097157-appb-100001
    其中X代表碳原子或氮原子；当X为氮原子时，R1无取代；当X为碳原子时，R1为氢、或C1～C4烷基；
    L代表-(CH2)n-、-CO(CH2)n-或-SO2(CH2)n-，其中n代表0～2；
    R2代表氢、甲基、C1～C4烷氧基、-NRaRb、
    Figure PCTCN2015097157-appb-100002
    Figure PCTCN2015097157-appb-100003
    m代表0或1；Ar代表苯环及或5～6员芳杂环；R4代表氢、卤素、氰基、硝基、甲氧基、C1～C4烷基、-NRaRb或-CH2-NRaRb；
    上述-NRaRb中的Ra、Rb各自独立代表氢、C1～C4烷基、C1～C4羟烷基或Ra、Rb连接形成5～6员含氮杂环。
  2. 权利要求1的化合物或其药学上可接受的盐，其中X为碳原子；R1代表甲基。
  3. 权利要求1的化合物或其药学上可接受的盐，其中L代表键、-CH2-、-CO-、-COCH2-或-SO2-。
  4. 权利要求1的化合物或其药学上可接受的盐，其中R2代表氢、-NRaRb、
    Figure PCTCN2015097157-appb-100004
    Figure PCTCN2015097157-appb-100005
    其中Ra、Rb、m、R4定义同权利要求1。
  5. 权利要求4的化合物或其药学上可接受的盐，其中R2代表-NRaRb,Ra、Rb各自独立代表氢、甲基、乙基、丙基、异丙基、正丁基；或者Ra、Rb连接形成四氢吡咯基、哌啶基、吗啡啉基、哌嗪基或N-甲基哌嗪基。
  6. 权利要求1的通式(I)化合物的制备方法，包括：
    Figure PCTCN2015097157-appb-100006
    其中X、R1、R2、L的定义如同权利要求1。
  7. 一种药物组合物，其中含有权利要求1的化合物或其药学上可接受的盐及药学上可接受的载体。
  8. 权利要求1的化合物或其药学上可接受的盐用于制备治疗恶性肿瘤的药物的用途。
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