WO2017133464A1 - 一种芳基苯并呋喃类酰胺化衍生物及医药用途 - Google Patents

一种芳基苯并呋喃类酰胺化衍生物及医药用途 Download PDF

Info

Publication number
WO2017133464A1
WO2017133464A1 PCT/CN2017/071614 CN2017071614W WO2017133464A1 WO 2017133464 A1 WO2017133464 A1 WO 2017133464A1 CN 2017071614 W CN2017071614 W CN 2017071614W WO 2017133464 A1 WO2017133464 A1 WO 2017133464A1
Authority
WO
WIPO (PCT)
Prior art keywords
halogen
substituted
unsubstituted
hydroxy
arylbenzofuran
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/CN2017/071614
Other languages
English (en)
French (fr)
Inventor
陈君
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
China Pharmaceutical University
Original Assignee
China Pharmaceutical University
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 China Pharmaceutical University filed Critical China Pharmaceutical University
Priority to US16/075,370 priority Critical patent/US11021454B2/en
Publication of WO2017133464A1 publication Critical patent/WO2017133464A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
    • C07D307/79—Benzo [b] furans; Hydrogenated benzo [b] furans with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring
    • C07D307/80—Radicals substituted by oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/06—Antigout agents, e.g. antihyperuricemic or uricosuric agents
    • 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
    • C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
    • C07D307/86—Benzo [b] furans; Hydrogenated benzo [b] furans with an oxygen atom directly attached in position 7
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism

Definitions

  • the invention belongs to the field of medicinal chemistry, in particular to an aryl benzofuran amidated derivative and a preparation method thereof, the derivative having anti-oxidation activity, the derivative having xanthine oxidase inhibitory activity, the derivative capable of It is used for preparing compositions, medicines and health care products for treating or preventing gout and hyperuricemia.
  • Gout is a kind of crystalline arthritis caused by the disorder of sputum metabolism and/or uric acid excretion in biological organisms.
  • the main clinical manifestations are hyperuricemia and characteristic acute arthritis and tophi, caused by urate crystal deposition.
  • Toxic chronic arthritis and can occur urate kidney disease, urinary acid urinary tract stones and other diseases (Wang Yu, China Pharmacy, 2009, 20 (22): 1748-1750).
  • Hyperuricemia is an important physiological and biochemical basis for inducing gout.
  • the main manifestation is that the serum uric acid concentration is too high.
  • the main reasons for its production are two aspects: one is excessive production of uric acid, and the other is reduced uric acid excretion.
  • Persistent hyperuricemia is a precursor and a potential cause of gout. Therefore, hyperuricemia in patients with gout plays a pivotal role in the treatment of gout (Wu Xinrong, Chinese Pharmacological Bulletin, 2010, 26(11): 1414-1417).
  • Xanthine Oxidase (XOD) is a key enzyme in the synthesis of uric acid and is one of the important drug targets for the treatment of hyperuricemia.
  • XOD is a complex flavinase, composed of two identical subunits, present in a variety of organisms and is an essential enzyme in the metabolism of nucleic acids in the body. It is widely distributed in the cytoplasm of human heart, liver, lung and other tissues. It can catalyze the formation of jaundice in hypoxanthine and further catalyze the oxidation of xanthine to produce uric acid and superoxide anion (Zhou Da, Food Science and Technology, 2009, 34(6) :174-178). Before the listing of febuxostat, allopurinol is the only drug that can be used clinically to inhibit the production of uric acid, but it has toxic side effects such as liver and kidney function damage.
  • Febuxostat is a new anti-gout drug approved by the US FDA for long-term control of hyperuricemia in patients with gout. It is a highly selective xanthine oxidase inhibitor that does not act on other related enzymes in the purine and pyrimidine metabolic pathways, and does not affect the normal metabolism of purines and pyrimidines in the body, but it also has abnormal liver function, joint pain, etc. Adverse reactions occur, which also affects its clinical application to some extent (Zhang Jing, Chinese Journal of Pharmaceutical Sciences, 2010, 45(15): 1197-1198). Therefore, the discovery and search for new safe and effective xanthine oxidase inhibitors are of great significance for the treatment of hyperuricemia and gout and other related diseases and human metabolism research.
  • Natural products have always been an important source of drugs for the treatment of various diseases, such as ligustrazine, sodium ferulate, vinpocetine and other cardiovascular drugs, podophyllotoxin, paclitaxel and other anti-tumor drugs are derived from natural products.
  • some natural products have poor water solubility, large toxic and side effects, rapid metabolism in the body, short biological half-life, etc., but after structural optimization, they have become first-line drugs for clinical use, such as sodium tanshinone IIA sulfonate, etoposide, polyene. Paclitaxel and other drugs. Therefore, proper structural modification of the active ingredients in natural products is an important way to develop new drugs.
  • Salvia miltiorrhiza is the dry root and rhizome of Salvia miltiorrhiza Bge., which is distributed in most parts of the country. Salvia miltiorrhiza has many functions such as promoting blood circulation to remove blood stasis, regulating menstruation and relieving pain, clearing heat and soothing the nerves. It is a blood-activating and stasis-relieving medicine commonly used by traditional Chinese medicine. For chest pain, heartache, abdominal pain, accumulation of symptoms, fever, pain, upset, irregular menstruation, pain After menstruation, sore throat and pain (People's Republic of China Pharmacopoeia (2010 edition)).
  • Danshen preparation has significant curative effect on many diseases, and is widely used in the treatment of coronary heart disease, hypercholesterolemia, hypertension, arrhythmia, hepatitis, peripheral vascular disease, pulmonary vascular disease, cirrhosis and other diseases (Wu Hao, Journal of Shenyang Pharmaceutical University, 2006, 23(1): 60-64). According to the nature of the chemical constituents of Salvia miltiorrhiza, it can be divided into two main categories: water-soluble components and fat-soluble components.
  • the fat-soluble components are mainly concentrated in diterpenoids represented by tanshinone, and the water-soluble components have a phenolic acid structure, which is one of the main material foundations for Danshen to exert good pharmacological effects, and is in chemical composition and medicine.
  • Generational dynamics and pharmacology have been extensively studied (Zhang Weiwei, Chinese Journal of Traditional Chinese Medicine, 2010, 35(3): 389-392).
  • Studies have shown that salvianolic acid components exert pharmacological effects through various mechanisms of action, such as inhibiting platelet aggregation, antithrombotic formation, preventing atherosclerotic plaque formation, inhibiting endogenous cholesterol synthesis, scavenging free radicals, and reducing freedom.
  • the base protects the body from damage (Du Guanhua, Basic Medicine and Clinical Medicine, 2000, 20(5): 10-14).
  • the water-soluble component originally isolated from danshensu ( ⁇ -3,4-dihydroxyphenyllactic acid) is the basic chemical structure of various salvianolic acids (Du Guanhua, Basic Medicine and Clinical Medicine, 2000, 20(5): 10 -14), such as salvianolic acid A is formed by the condensation of one molecule of Danshensu with two molecules of caffeic acid, salvianolic acid B is a mixture of three molecules of Danshensu and one molecule of caffeic acid, and salvianolic acid C consists of two molecules. Danshensu is condensed.
  • phenolic compounds in Salvia miltiorrhiza also include salvianolic acid D, salvianolic acid E, salvianolic acid F, salvianolic acid G, salvianolic acid H, salvianolic acid I, rosmarinic acid, lithosperic acid and the like.
  • salvianolic acid B, lithospermic acid, rosmarinic acid and danshensu in salvia miltiorrhiza have jaundice oxidase inhibition and can reduce serum uric acid levels in hyperuricemia mice (CN) 200410084620.4).
  • Salvianolic acid C has strong xanthine oxidase inhibitory activity and its inhibitory effect is superior to salvianolic acid A (Yan Yuting, Journal of China Pharmaceutical University, 2013, 44(5): 442 -446). Salvianolic acid C can be used as a xanthine oxidase inhibitor for the prevention and treatment of gout and hyperuricemia and its complications (CN201210000772.6).
  • salvianolic acid C also has a structural skeleton of an arylbenzofuran.
  • Arylbenzofurans have long been a hot spot for researchers. It is a class of compounds with a new lignin skeleton type, distributed in a variety of higher plants in nature, and has a wide range of biological activities, such as antiviral, antitumor, antifungal, antioxidant, immunomodulatory and cardiovascular diseases. Wait. In recent years, the research progress of arylbenzofuran compounds has been reviewed.
  • salvianolic acid C is highly water-soluble, sensitive to air, strong acid, strong alkali, etc., physical and chemical stability is poor, and the process in the body is easily metabolized, and the biological half-life is short, and its clinical application has certain limitations. Therefore, we designed and synthesized a series of arylbenzofuran derivatives with the target of the 2-arylbenzofuran ring contained in the structure of salvianolic acid C, and related biological activities. In-depth discussion has certain research value.
  • the invention discloses an arylbenzofuran amidated derivative.
  • the present invention discloses an antioxidant activity of an arylbenzofuran amidated derivative.
  • the invention discloses an activity of scavenging free radicals of an arylbenzofuran amidated derivative.
  • the present invention discloses an arylbenzofuran amidated derivative having the inhibitory activity of xanthine oxidase.
  • the present invention discloses an arylbenzofuran amidated derivative having an activity of reducing uric acid.
  • the present invention discloses an arylbenzofuran amidated derivative having activity for treating or preventing gout.
  • the present invention discloses an arylbenzofuran amidated derivative for use in the preparation of a composition, a medicament, a health care product for treating or preventing gout, and or hyperuricemia.
  • the invention discloses an arylbenzofuran amidated derivative for preparing a composition, a medicament and a health care product for treating or preventing gout and hyperuricemia.
  • the invention discloses a preparation method of an arylbenzofuran amidated derivative.
  • the invention discloses an arylbenzofuran amidated derivative.
  • the invention discloses an arylbenzofuran amidated derivative, the structural formula of which is as shown in formula I:
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 are each independently hydrogen, hydroxy, halogen, nitro, benzyl, unsubstituted or halogen, hydroxy, nitro, C One to three substituent-substituted C 1-4 alkyl groups in the 1-2 alkoxy group, unsubstituted or one to two substituents selected from halogen, hydroxy, nitro, and C 1-2 alkoxy groups a substituted C 1-3 alkoxy group;
  • R 10 is hydrogen, —COOH, —COOCH 3 , —COOCH 2 CH 3 , —COOCH(CH 3 ) 2 , —COO(CH 2 ) 2 CH 3 or via halogen, hydroxyl, C 1-3 alkyl, C 1 Among the -2 alkoxy groups, one to three substituent-substituted C 1-4 alkyl groups are selected.
  • the present invention discloses an arylbenzofuran amidated derivative of the formula I, which is characterized in that:
  • R 3 , or , and R 4 are other substituents other than a hydroxyl group
  • R 9 , or , and R 10 are other substituents other than hydrogen.
  • the invention discloses a class of aryl benzofuran amidated derivatives, and the structural formula is as shown in formula II:
  • the invention discloses a class of aryl benzofuran amidated derivatives, the structural formula is as shown in formula III:
  • the invention discloses a class of aryl benzofuran amidated derivatives, and the structural formula is as shown in formula IV:
  • the invention discloses a class of aryl benzofuran amidated derivatives, and the structural formula is as shown in formula V:
  • the invention discloses a class of aryl benzofuran amidated derivatives, and the structural formula is as shown in formula VI:
  • the invention discloses a class of aryl benzofuran amidated derivatives, the structural formula is as shown in formula VII:
  • the invention discloses a class of aryl benzofuran amidated derivatives, the structural formula is as shown in formula VIII:
  • the invention discloses a class of aryl benzofuran amidated derivatives, and the structural formula is as shown in formula IX:
  • the invention discloses a class of aryl benzofuran amidated derivatives, the structural formula is as shown in formula X:
  • the invention discloses a class of aryl benzofuran amidated derivatives, and the structural formula is as shown in formula XI:
  • the invention discloses a class of aryl benzofuran amidated derivatives, the structural formula is as shown in formula XII:
  • the invention discloses a class of aryl benzofuran amidated derivatives, the structural formula is as shown in formula XIII:
  • R 1 , R 2 , and R 3 are each independently a C 1-4 alkyl group which is unsubstituted or substituted with a halogen or a hydroxy group, a C 1-3 alkoxy group which is unsubstituted or substituted with a halogen or a hydroxy group, a halogen, a nitro group, Benzyl;
  • R 5 is hydrogen, -COOH, -COOCH 3 , -COOCH 2 CH 3 , -COOCH(CH 3 ) 2 , -COO(CH 2 ) 2 CH 3 or hydrogen, C 1-4 alkyl.
  • the invention discloses an arylbenzofuran amidated derivative of the formula 1, which is characterized in that:
  • Equation 1 The structural formula is as shown in Equation 1:
  • R 1 , R 2 and R 3 are each independently hydrogen and hydroxy; R 4 is hydrogen; and R 5 is hydrogen.
  • the present invention discloses an arylbenzofuran amidated derivative represented by Formula 1, which is characterized in that:
  • R 1 , R 2 , and R 3 are each independently a C 1-4 alkyl group which is unsubstituted or substituted with a halogen or a hydroxy group, a C 1-3 alkoxy group which is unsubstituted or substituted with a halogen or a hydroxy group, a halogen, a nitro group, Benzyl;
  • R 5 is -COOH, -COOCH 3 , -COOCH 2 CH 3 , -COOCH(CH 3 ) 2 ,
  • —COO(CH 2 ) 2 CH 3 is either hydrogen or C 1-4 alkyl.
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 5a:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 6a:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 5b:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 6b:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 5c:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 6c:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 5d:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 6d:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 5e:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 6e:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 5f:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in formula 6f:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in the formula:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in 9a:
  • the invention discloses an arylbenzofuran amidated derivative, and the structural formula is as shown in 10a:
  • the salvianolic acid C is mixed with an inorganic base, ultrasonically dissolved in a mixed solvent, and the reaction is continuously detected by heating. After the reaction is completed, the silica gel column chromatography is carried out to obtain an intermediate Tournefolic acid A (2);
  • the inorganic base hydrolysis in the first step may be selected from LiOH, NaOH, KOH, etc., and the organic solvent is THF, MeOH, H 2 O or a mixed solvent thereof, preferably a mixed solvent MeOH/H 2 O, and the solvent volume ratio It is preferably 3:1 to 5:1, and the reaction time is preferably 8 to 12 hours.
  • the organic solvent is THF, MeOH, H 2 O or a mixed solvent thereof, preferably a mixed solvent MeOH/H 2 O, and the solvent volume ratio It is preferably 3:1 to 5:1, and the reaction time is preferably 8 to 12 hours.
  • the separation and purification method in the first step is specifically: silica gel column chromatography, eluting with chloroform/methanol/formic acid (10:1:0.1, v/v/v) to obtain the intermediate Tournefolic acid A (2) ).
  • the amino compound in the second step is an amino acid of various D/L configurations and other chiral amino derivatives
  • the solvent is a reagent such as methanol, ethanol or isopropanol, and the reaction time is preferably 18 to 24 hours.
  • the separation and purification method in the second step is specifically: washing the sample by multiple interactions of methanol and diethyl ether, respectively, and concentrating and evaporating.
  • the condensing agent in the third step may be DCC/HoBt, EDCI/HoBt, HATU, HBTU, PyBOP, preferably EDCI/HoBt, and the organic solvent is preferably a mixed solvent DMF/CH 2 Cl 2 (3:1 ⁇ 5) :1v/v), the reaction time is preferably 8 to 12 hours.
  • the separation and purification method in the third step is specifically: silica gel column chromatography, and a chloroform/methanol gradient elution to obtain a target derivative.
  • the inorganic base in the fourth step may be LiOH, NaOH, KOH, preferably NaOH, and the organic solvent is THF, MeOH, H 2 O or a mixed solvent thereof, preferably MeOH/H 2 O (3:1 ⁇ 5:1v/v), the reaction time is preferably 8 to 12 hours.
  • the specific separation and purification method of the partial arylbenzofuran amidated derivative in the fourth step is: separation and purification by silica gel column chromatography, the mobile phase is chloroform/methanol/formic acid gradient elution; or high-performance liquid phase preparation
  • the sample mixture is separated and purified by a column, wherein the column (Agilent, Zorbax-C 18 , 5 ⁇ m, 9.4 ⁇ 250 mm), the chromatographic conditions are preferably: flow rate: 8 mL/min, detection wavelength: 281 nm, column temperature: 30 ° C, mobile phase : acetonitrile - 0.1% formic acid - water.
  • the compounds obtained in the preparation method were identified by spectroscopic methods such as mass spectrometry and nuclear magnetic resonance, and the purity was detected by HPLC.
  • the arylbenzofuran amidated derivative of the present invention and a pharmaceutically acceptable salified product thereof can be combined with a common medicinal excipient and a carrier to prepare a composition of a gout and hyperuricemia drug, thereby achieving prevention or The effect of treatment.
  • the above-mentioned drugs can be selected according to actual needs, such as tablets, injections, suppositories, aerosols, nano preparations and the like.
  • the present invention has the following beneficial effects:
  • the present invention obtains a series of arylbenzofuran derivatives by chemically modifying the salvianolic acid C as a substrate.
  • the in vitro bioactive screening and cell viability assay showed that the arylbenzofuran amidated derivatives of the present invention exhibited strong xanthine oxidase inhibitory activity and antioxidant activity as a whole, and were gout and hyperuricemia.
  • the prevention and treatment of its complications provide a material basis.
  • the arylbenzofuran amidated derivative of the present invention has a novel structure and has a good inhibitory effect on xanthine oxidase. Compared with salvianolic acid C, the water solubility is improved to some extent, and the physicochemical stability of the compound is increased to some extent, and the toxic and side effects are low, which has certain development and application value.
  • Figure 1 Scavenging effects of arylbenzofuran amidated derivatives 5a, 5b, 5c, 6b, 6c, 6d, 6e and Salvianolic acid C(1) and Ve on LPS-stimulated macrophage RAW 264.7 superoxide anion
  • Figure 3 is a detailed explanation: Figure 3 is a combination of Part A, Part B, Part C, Part D, Part E, and Part F, showing the results of docking experiments between different compounds and xanthine oxidase molecules, (A) Febuxostat ( B) Allopurinol (C) 6b (D) 6e (E) Salvianolic acid C (1) (F) 5b.
  • the reagents were purchased from Sigma-Aldrich or Aladdin Reagent, the reagents were of analytical grade or chemical purity, and the deuterated reagents were purchased from CIL Reagent, Cambridge, USA.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • Tournefolic acid A can be identified as C 17 H 12 O 6 , and its structure is as follows.
  • the structure of the reaction product obtained in the step (2) was subjected to structural analysis.
  • the compound 4a can be identified as having a chemical formula of C 10 H 14 ClNO 4 having the following formula.
  • Tournefolic acid A (36 mg, 0.115 mmol) was accurately weighed in a round bottom flask, DMF/CH 2 Cl 2 4 mL (4:1 v/v) was added, sonicated, cooled in an ice bath, and accurately weighed EDCI (33.1 mg) , 0.173 mmol), HoBt (23.3 mg, 0.173 mmol), cooled in ice bath for 10 min and stirring.
  • the structure of the reaction product obtained in the step (3) was subjected to structural analysis.
  • the sample mixture was separated by preparative liquid chromatography (Agilent zorbax-C 18 , 5 ⁇ m, 20 ⁇ 250 mm) at a flow rate of 8 mL/min, a detection wavelength of 281 nm, and a mobile phase: acetonitrile-0.1% formic acid-water to obtain the target product 6a (11.6 mg). , yield 66%).
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • Tourugolic acid A 32 mg, 0.103 mmol
  • DMF/CH 2 Cl 2 3.5 mL 5:1 v/v
  • sonicate to dissolve the sample, cool in ice bath, and add accurately weighed EDCI.
  • HoBt (22.3 mg, 0.165 mmol)
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • Thornefolic acid A (35 mg, 0.112 mmol) was accurately weighed into a round bottom flask, DMF/CH 2 Cl 2 4 mL (3:1 v/v) was added, ultrasonicated until the sample dissolved, cooled to 0 ° C in an ice bath, and EDCI (32.3) was added. Mg, 0.168 mmol), HoBt (22.6 mg, 0.168 mmol), cooled in ice bath for 10 min and stirring.
  • the structure of the reaction product obtained in the step (3) was subjected to structural analysis.
  • the chemical formula of the compound 5c can be identified as C 21 H 19 NO 7 , and its structural formula is as follows.
  • reaction mixture was cooled in an ice bath, and a 10% aqueous HCl solution was added dropwise thereto, and the mixture was stirred until the pH was 3 to 4, and the MeOH was evaporated to dryness, and 10 mL of distilled water was added thereto, and extracted with ethyl acetate (10 mL ⁇ 4), and organic The layer was concentrated, washed with saturated brine, dried over MgSO 4 and then evaporated.
  • the mixture was purified by preparative liquid chromatography (Agilent zorbax-C 18 , 5 ⁇ m, 20 ⁇ 250 mm), flow rate 8 mL/min, detection wavelength 281 nm, mobile phase: acetonitrile- 0.1% formic acid-water gave the derivative 6c (13.1 mg, yield 71%).
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the chemical formula of the compound 6c can be identified as C 20 H 17 NO 7 , and its structural formula is as follows.
  • Tournefolic acid A (35 mg, 0.112 mmol) was accurately weighed into a 25 mL round bottom flask, DMF/CH 2 Cl 2 3 mL (4:1 v/v) was added, sonicated until the sample was dissolved, cooled in an ice bath, and EDCI (30.2 mg, 0.157 mmol), HoBt (21.2 mg, 0.157 mmol), cooled in ice bath for 10 min and stirring.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the chemical formula 5d can be identified as C 27 H 23 NO 7 , and its structural formula is as follows.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the chemical formula of the compound 6d can be identified as C 26 H 21 NO 7 , and its structural formula is as follows.
  • Tournefolic acid A (33 mg, 0.106 mmol) was accurately weighed into a round bottom flask, DMF/CH 2 Cl 2 4 mL (3:1 v/v) was added, and the sample was completely dissolved by ultrasonication, cooled in an ice bath, and EDCI (30.4 mg, 0.158 mmol), HoBt (21.5 mg, 0.159 mmol), cooled in ice bath for 10 min and stirring.
  • Add L-tyrosine methyl ester hydrochloride (29.4 mg, 0.127 mmol), Et 3 N (32.1 mg, 0.318 mmol), EtOAc (EtOAc) m. :1) The reaction is continuously monitored until the raw materials are substantially eliminated.
  • the structure of the reaction product obtained in the step (3) was subjected to structural analysis.
  • the chemical formula of the compound 5e can be identified as C 27 H 23 NO 8 , and its structural formula is as follows.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the chemical formula of the compound 6e can be identified as C 26 H 21 NO 8 , and its structural formula is as follows.
  • Tournefolic acid A (41mg, 0.131mmol) in a round bottom flask, add DMF / CH 2 Cl 2 5mL (5:1v / v), ultrasonically until the sample is completely dissolved, cooled to 0 ° C in an ice bath, added accurately The amount of EDCI (37.8 mg, 0.197 mmol), HoBt (26.5 mg, 0.196 mmol) was cooled in ice bath for 10 min and stirring.
  • the structure of the reaction product obtained in the step (3) was subjected to structural analysis.
  • the sample mixture was separated by preparative liquid chromatography (Agilent zorbax-C 18 , 5 ⁇ m, 20 ⁇ 250 mm) at a flow rate of 8 mL/min, a detection wavelength of 281 nm, and a mobile phase: acetonitrile-0.1% formic acid-water to obtain a reaction product 6f (10.7 mg). , yield 55%).
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the compound 9a can be identified as having a chemical formula of C 30 H 29 NO 9 having the following structure.
  • the sample was separated by preparative liquid chromatography (Agilent zorbax-C 18 , 5 ⁇ m, 20 ⁇ 250 mm) at a flow rate of 8 mL/min, a detection wavelength of 281 nm, and a mobile phase: acetonitrile-0.1% formic acid-water to obtain a reaction product 10a (10.7 mg, Yield 55%).
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the chemical formula of the compound 10a can be identified as C 29 H 27 NO 9 , and its structure is as follows.
  • the structure of the reaction product obtained in the step (1) was subjected to structural analysis.
  • the compound 9b can be identified as having a chemical formula of C 30 H 29 NO 9 having the following structure.
  • (2) XOD solution Take 25U/2.6mL of XOD, dilute to 0.08U/mL of XOD working solution with 75mM PB solution, mix well with pipette, store on ice, set aside;
  • Substrate preparation accurately weigh the appropriate amount of xanthine (XA) into 5mL of 0.1N NaOH solution, ultrasonically dissolve, the latter added 95mL of 75mM PB solution to prepare the final mother liquor of 0.48mM concentration, vortex mixed Evenly 1 min, freshly prepared before each experiment;
  • test drug accurately weigh the appropriate amount of the test drug, dissolved in DMSO to prepare a 10 mM stock solution, stored at -20 ° C in the dark. DB was diluted to different concentrations (0-100 mM) before the experiment, and the DMSO content was less than 0.1%.
  • IC 50 value is the average of four parallel experiments
  • the antioxidant activity of the synthesized benzofuran amidated derivative was evaluated by DPPH free radical scavenging experiment.
  • test drugs accurately weigh the appropriate amount of test drug, dissolve 10m stock solution with MeOH, and store at -20 °C in the dark. Dilute to different concentrations (0-100 mM) with MeOH before the experiment.
  • IC 50 values are the average results of three parallel experiments.
  • the antioxidant activity evaluation (the ability to scavenge superoxide anion) of the synthesized benzofuran amidated derivative was evaluated by a cell model.
  • LPS Lipopolysaccharide
  • the present invention utilizes the LPS-stimulated macrophage model to evaluate the antioxidant activity of benzofuran amidated derivatives at the cellular level to scavenge superoxide anion.
  • Drug to be tested accurately weigh the appropriate amount of the test drug, prepare a 10 mM sample stock solution in DMSO, and store at -20 ° C for use.
  • LPS accurately weigh the appropriate amount of LPS, and prepare a sample stock solution of 0.1 ⁇ g/mL in DMEM and store at -20 °C for use.
  • Probe Stabilizer DTPA Weigh accurately the amount of DTPA, prepare a 20 mM sample stock solution in DMSO, dilute to 100 ⁇ M with PB, and store at 4 ° C until use.
  • RAW 264.7 cells were seeded in a Petri dish (60 mm), 4 mL per dish, and placed in a saturated humidity, 37 ° C 5% CO 2 incubator. The experiment was divided into blank group, model group and drug-administered group. The model group and the drug-administered group were respectively added with the molding agent LPS (prepared in DMEM). The optimal LPS concentration was 0.1 ⁇ g/mL, and the modeling time was 24 hours. Vitamin E is a positive control.
  • test drug having a final concentration of 10 ⁇ M in a 0.1 ⁇ g/mL LPS medium was added, and two parallel wells were set for each sample to be tested.
  • the cells were then washed twice with ice-cold PBS/(100 ⁇ M) DTPA, then 1 mL PBS/(100 ⁇ M) DTPA was added, and the cells were separately divided into two portions (700 ⁇ L and 300 ⁇ L) with a spatula and collected into EP tubes at 4 ° C. After centrifugation at 13,000 rpm for 10 min, the supernatant was removed, and the cell pellet was stored at -80 ° C until use.
  • the former contained 700 ⁇ L of cell liquid for the determination of the LC-MS content of the test substance 2-OH-E + , and the latter contained 300 ⁇ L of the cell liquid for the determination of the cell protein concentration.
  • LC-MS chromatographic conditions instrument: Agilent 1100 series LC-QMS; column: Agilent Zorbax SB-Aq (4.6 x 50 mm, 1.8 ⁇ m); mobile phase: 0.1% formic acid-water (phase A); 0.1% formic acid - acetonitrile (B); Gradient conditions: 0 to 2 min, 5% to 30% B; 2 to 5 min, 30% B; 5 to 7 min, 30% to 100% B; 7 to 10 min, 100% B; flow rate: 1 mL /min; MS conditions: positive ion mode, SIM (SIM1: 1.2 ⁇ 3min [M + H] + 288 (galantamine); 3 ⁇ 10min [M] + 330 (2-OH-E + )); Voltage: 120V; nitrogen flow rate: 10.0L/min;
  • test drug accurately weigh the appropriate amount of the test drug, prepare a 10 mM sample stock solution in DMSO, and store it at 4 ° C for use.
  • test drug was diluted with the medium to the required concentration (10 ⁇ M, 30 ⁇ M, 50 ⁇ M), and then added to a 6-well plate. After incubation at 37 ° C for 15 min, the model group and the drug-administered group were respectively A total of 10 ⁇ M of Astragalus membranaceus was added, and two duplicate wells were set in each group. After being cultured for 24 hours in a cell culture incubator, 100 ⁇ L of the culture medium was collected for LC-MS analysis.
  • test drug accurately weigh the appropriate amount of the test drug, dissolve it with the corresponding volume of physiological saline, clarify the solution, prepare it as a stock solution, and store it at 4 °C for use.
  • mice were randomly divided into 9 groups, 10 in each group: 1 normal control group, 2 model control group, 3 derivatives 5b low dose group: 20 mg/kg, 4 derivatives 5b medium dose group: 40 mg /kg,5 derivative 5b high dose group: 80mg/kg, 6 derivative 6e low dose group: 20mg/kg, 7 derivative 6e medium dose group: 40mg/kg, 8 derivative 6e high dose group: 80mg/kg , 9 positive control group: allopurinol 20mg/kg.
  • the test drug was prepared with 0.5% sodium carboxymethylcellulose (CMC-Na) to prepare a suitable concentration.
  • mice in the normal group and the model group were intraperitoneally injected with 0.5% sodium carboxymethylcellulose, and the other groups were intraperitoneally injected with the test drug 10 mL/kg.
  • 500 ⁇ L of blood was taken from the eyeball, placed at room temperature for 1 h, centrifuged at 3000 rpm for 5 min, and the upper serum was stored at -4 °C.
  • the uric acid level in the serum of the mice was determined according to the method of the uric acid detection kit.
  • the liver of the mouse was taken, rinsed with physiological saline, 100 mg, and 900 ⁇ L of pre-cooled physiological saline was added to homogenate, centrifuged, and the supernatant was taken to determine the activity of xanthine oxidase in the liver homogenate according to the method of the kit.
  • Xanthine oxidase (PDB code: 1FIQ) consists of three subunits A, B, and C, which contain three active domains, which are the Fe/S central domain in the A chain, and the FAD domain in the B chain. Molybdenum pterin domain in the C chain.
  • compounds 1, 6e, 6b and Febuxostat interact with multiple identical amino acid residues, such as Lys249, Lys256, Leu257, Val259, Ile264, Ile353, Arg394 and Leu404, indicating compound 1,6e, 6b may be similar to the positive drug Febuxostat, and binds to the same domain of xanthine oxidase to exert an activity of inhibiting the enzyme. It can also be seen from Fig.
  • the derivative 5b interacts with the amino acid residues Pro1076, Phe1009, Thr1010, Arg880, Val1011, Leu1014, Glu802, Ser876, Glu879, and forms three hydrogens with the residues Phe1009, Thr1010and Glu879.
  • the bond which is similar to the positive drug Allopurinol, may act on the common domain of xanthine oxidase and exert an enzyme inhibitory effect. It has also been observed that the formation of hydrogen bonds and hydrophobic interactions play an important role in stabilizing the conformation of the receptor-ligand ligand.
  • Preparation process Take the derivative 5b of the present invention through a 100 mesh sieve, add starch and magnesium stearate to mix uniformly, make granules, dry and compress, and obtain.
  • Preparation process taking the derivative 5b of the present invention through a 100 mesh sieve, adding starch and magnesium stearate to form a uniform
  • Granules dry, capsules, that's it.
  • Preparation process take the derivative 5b of the invention, add soybean soft phospholipid, colloid mill and mix, vacuum, press, that is, soft capsule.
  • Derivative 5b2g sodium sulfite 4g, ethanol 50mL, add water to a volume of 1000mL;
  • Preparation process Take the derivative 5b of the invention and disperse it in ethanol, dissolve the sodium sulfite in water, and gradually add the sodium sulfate solution under ultrasonic or stirring to make a clear and transparent solution; make up the water to a sufficient amount; The ⁇ m microporous membrane was filtered and lyophilized.
  • Preparation process Take the derivative 6e of the present invention through a 100 mesh sieve, add starch and magnesium stearate to mix uniformly, make granules, dry and compress, and obtain.
  • Preparation process taking the derivative 6e of the present invention through a 100 mesh sieve, adding starch and magnesium stearate to form a uniform mixture
  • Granules dry, capsules, that's it.
  • Preparation process taking the derivative 6e of the invention, adding soybean soft phospholipid, colloidal grinding and mixing, vacuuming, pressing, that is, obtaining a soft capsule.
  • Derivative 6e2g sodium sulfite 4g, ethanol 50mL, add water to a volume of 1000mL;
  • Preparation process Take the derivative 6e of the invention and disperse it in ethanol, dissolve the sodium sulfite in water, and gradually add the sodium sulfate solution under ultrasonic or stirring to make a clear and transparent solution; make up the water to a sufficient amount; The ⁇ m microporous membrane was filtered and lyophilized.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Rheumatology (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pain & Pain Management (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明公开了一种芳基苯并呋喃类酰胺化衍生物及医药用途、制备方法，该衍生物具备抗氧化的活性，具备黄嘌呤氧化酶抑制活性，能用于制备抗氧化、治疗痛风及高尿酸血症的组合物、药物、保健品。

Description

一种芳基苯并呋喃类酰胺化衍生物及医药用途 技术领域
本发明属于药物化学领域，特别涉及一种芳基苯并呋喃类酰胺化衍生物及其制备方法，该衍生物具备抗氧化的活性，该衍生物具备黄嘌呤氧化酶抑制活性，该衍生物能用于制备治疗或预防痛风及高尿酸血症的组合物、药物、保健品。
技术背景
随着人们生活水平的不断提高，生活方式及饮食结构也发生了很大变化，过多的摄入嘌呤和蛋白质饮食使高尿酸血症和痛风的患病率正呈逐渐上升的趋势，已成为一种常见病、多发病。痛风常与其他代谢性疾病相伴发生，如冠心病、高血压、高血脂、糖尿病等，给人类生命健康和财产带来严重危害(张建明，郑州大学学报，2009，44(1)：174)。痛风，作为一种严重影响公共健康的代谢性疾病，在我国，尤其是南方地区，发病率正呈现逐步上升的趋势，其预防工作也不断引起人们的重视，安全有效的治疗方法与药物迫在眉睫。痛风是由生物机体内嘌呤代谢紊乱和/或尿酸排泄减少引起的一种晶体性关节炎，临床主要表现为高尿酸血症和尿酸盐结晶沉积所致的特征性急性关节炎、痛风石、痛风石性慢性关节炎，并可发生尿酸盐肾病、尿酸性尿路结石等病症(王宇，中国药房，2009，20(22)：1748-1750)。
高尿酸血症是诱发痛风的重要生理生化基础，主要表现为血清尿酸浓度过高，其产生的主要原因有两个方面：一是尿酸生成过多，二是尿酸排泄减少。持续性的高尿酸血症是痛风的前兆和潜在的诱因。因此，治疗痛风患者的高尿酸血症在治疗痛风过程中有着举足轻重的作用(吴新荣，中国药理学通报，2010，26(11)：1414-1417)。然而，黄嘌呤氧化酶(Xanthine Oxidase，简称XOD)是尿酸合成过程中的关键酶，是治疗高尿酸血症的重要药物靶点之一。XOD是一种复合黄素酶，由两个相同的亚基组成，存在于多种生物体内，是机体内核酸代谢过程中至关重要的一种酶。它广泛分布于人体心、肝、肺等组织细胞浆中，可催化次黄嘌呤生成黄嘌呤，并进一步催化黄嘌呤氧化产生尿酸和超氧阴离子(周达，食品科技，2009，34(6)：174-178)。在非布索坦上市以前，别嘌醇(Allopurinol)是临床上应用的唯一能够抑制尿酸生成的药物，但其有肝肾功能损伤等毒副作用。而非布索坦(Febuxostat)是由美国FDA批准的抗痛风新药，可用于长期控制痛风患者的高尿酸血症。它是一种高选择性的黄嘌呤氧化酶抑制剂，不作用于嘌呤和嘧啶代谢途径中的其他相关酶，不影响机体内嘌呤与嘧啶的正常代谢，但其也有肝功能异常、关节疼痛等不良反应发生，这在一定程度上也影响了其临床应用(张静，中国药学杂志，2010，45(15)：1197-1198)。因此，发现和寻找新的安全有效的黄嘌呤氧化酶抑制剂对于高尿酸血症和痛风等相关疾病的治疗及人体代谢学研究等方面具有重要意义。
天然产物一直都是治疗各类疾病药物的重要源泉，如川芎嗪、阿魏酸钠、长春西汀等心血管药物，鬼臼毒素、紫杉醇等抗肿瘤药物均来自于天然产物。此外，一些天然产物具有水溶性差，毒副作用大，体内代谢快，生物半衰期短等缺陷，但经过结构优化后，也成为临床使用的一线药物，如丹参酮IIA磺酸钠、依托泊苷、多烯紫杉醇等药物。因此，对天然产物中的活性成分进行适当的结构修饰是新药研发的重要途径。
丹参为唇形科鼠尾草属植物丹参(Salvia miltiorrhiza Bge.)的干燥根及根茎，全国大部分地区都有分布。丹参具有活血化瘀、调经止痛、清热安神等多种功效,是中医常用的活血化瘀药。用于胸痹心痛，脘腹胁痛，癥瘕积聚，热痹疼痛，心烦不眠，月经不调，痛 经经闭，疮疡肿痛(中华人民共和国药典(2010年版))。丹参制剂对许多疾病都有着显著的疗效，临床上广泛用于治疗冠心病、高胆固醇血症、高血压、心律失常、肝炎、周围性血管疾病、肺血管疾病、肝硬化等疾病(吴浩，沈阳药科大学学报，2006，23(1)：60-64)。根据丹参化学成分的性质，主要可以分为两大类：水溶性成分和脂溶性成分。其中，脂溶性成分主要集中在以丹参酮为代表的二萜醌类化合物，而水溶性成分则多具有酚酸类结构，是丹参发挥良好药效的主要物质基础之一，并在化学成分、药代动力学及药理学等方面已被广泛研究(张伟伟，中国中药杂志，2010，35(3)：389-392)。研究表明，丹酚酸类成分通过多种作用机制而发挥药理作用，如抑制血小板聚集、抗血栓形成、防止动脉粥样硬化斑块形成、抑制细胞内源性胆固醇合成、清除自由基、减少自由基对机体的损伤而产生保护作用等(杜冠华，基础医学与临床，2000，20(5)：10-14)。
最初分离出的水溶性成分为丹参素(β-3,4-二羟基苯乳酸)是各种丹酚酸类化合物的基本化学结构(杜冠华，基础医学与临床，2000，20(5)：10-14)，如丹酚酸A是由一分子丹参素与两分子咖啡酸缩合而成，丹酚酸B为三分子丹参素与一分子咖啡酸缩合而成，而丹酚酸C由两分子丹参素缩合而成。丹参中其它酚酸类化合物还包括丹酚酸D、丹酚酸E、丹酚酸F、丹酚酸G、丹酚酸H、丹酚酸I、迷迭香酸、紫草酸等。朱大元等发现，丹参中的丹酚酸B、紫草酸、迷迭香酸和丹参素这些酚酸类物质具有黄嘌呤氧化酶抑制作用，并能降低高尿酸血症小鼠的血清尿酸水平(CN 200410084620.4)。
本课题组前期研究发现，丹酚酸C具有较强的黄嘌呤氧化酶抑制活性，且抑制作用明显优于丹酚酸A(燕玉婷，中国药科大学学报，2013，44(5)：442-446)。丹酚酸C可以作为黄嘌呤氧化酶抑制剂用于防治痛风和高尿酸血症及其并发症(CN201210000772.6)。
Figure PCTCN2017071614-appb-000001
燕玉婷等对丹酚酸C在大鼠体内的代谢过程进行了研究，运用HPLC-QTOF-MS/MS的方法从大鼠血液及尿液中检测得到五种代谢产物，并根据化合物裂解规律对丹酚酸C在大鼠体内的药物代谢途径进行了初步推测(燕玉婷，中国药科大学学报，2013，44(5)：442-446)。
Figure PCTCN2017071614-appb-000002
综上所述，可以推测：β-3,4-二羟基苯乳酸的结构单元与黄嘌呤氧化酶的抑制活性可能具有一定的相关性。但是，丹酚酸C还具有芳基苯并呋喃的结构骨架。芳基苯并呋喃类化合物，一直以来都是研究者们所关注的热点。它是一类具有新木质素骨架类型的化合物，分布于自然界多种高等植物中,且具有广泛的生物学活性，如抗病毒、抗肿瘤、抗真菌、抗氧化、免疫调节及心血管疾病方面等。综述近年来对芳基苯并呋喃类化合物的研究进展，其研究工作多集中在苯并呋喃类骨架化合物的分离鉴定、衍生物库的构建合成及构效关系的探讨、生物活性的筛选及作用机制的深入研究，尤其是构建苯并呋喃环新方法的不断发现(蒲文臣，2-芳基苯并呋喃衍生物的生物活性与合成策略，有机化学，2011，31，155-165)。
基于以上内容，仍有一些尚且未知的研究问题需要探讨解决。(1)芳基苯并呋喃环对具有此骨架类型的化合物(如丹酚酸C等)所表现出的生物活性(如黄嘌呤氧化酶抑制活性、抗氧化活性)具有怎样的影响；(2)不同取代基(如吸电子基、供电子基等)取代时对芳基苯并呋喃环类化合物的相关药理活性的影响暂不可知；即针对特定的生物活性，其具体的构效关系仍不明确；(3)本领域技术人员暂不能确定：不含有β-3,4-二羟基苯乳酸结构而仅含有芳基苯并呋喃类化合物是否也具备与β-3,4-二羟基苯乳酸类化合物类 似的生物活性，例如具备抑制黄嘌呤氧化酶的活性；同时，本领域技术人员也不能肯定：不含有β-3,4-二羟基苯乳酸结构而仅含有芳基苯并呋喃类酰胺化衍生物是否也具备与含有β-3,4-二羟基苯乳酸结构类化合物类似的生物活性，例如具备抑制黄嘌呤氧化酶的活性。
此外，丹酚酸C水溶性较强，对空气、强酸、强碱等较敏感，物理化学稳定性较差，且体内过程易被代谢，生物半衰期较短，其临床应用有一定局限性。因此，我们以丹酚酸C结构中所含的2-芳基苯并呋喃环的母核结构为目标，设计并合成了系列芳基苯并呋喃类衍生物，并对其相关的生物活性进行了深入探讨，具有一定的研究价值。
发明内容
本发明公开了一种芳基苯并呋喃类酰胺化衍生物。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物抗氧化的活性。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物清除自由基的活性。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物具备黄嘌呤氧化酶的抑制活性。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物具备降低尿酸的活性。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物具备治疗或预防痛风的活性。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物用于制备治疗或预防痛风、和、或高尿酸血症的组合物、药物、保健品。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物用于制备治疗或预防痛风及高尿酸血症的组合物、药物、保健品。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物的制备方法。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，其结构通式如式I所示：
Figure PCTCN2017071614-appb-000003
R1，R2，R3，R4，R5，R6，R7，R8各自独立为氢，羟基，卤素，硝基，苄基，未取代或经卤素、羟基、硝基、C1-2烷氧基中选出1～3个取代基取代的C1-4烷基，未取代或经卤素、羟基、硝基、C1-2烷氧基中选出1～2个取代基取代的C1-3烷氧基；
R9为氢，未取代或经卤素、羟基、巯基、C1-3烷基、C1-2烷氧基中选出1～3个取代基取代的C1-4烷基，未取代或经卤素、羟基取代的3-乙基-1H-吲哚，未取代或经卤素、羟基、C1-2烷氧基取代的苯基，未取代或经卤素、羟基、C1-2烷氧基取代的苄基，未取代或经卤素、羟基、C1-2烷氧基取代的其他芳基(其中，所述卤素X＝F，Cl，Br，n＝1，2，3)，如3,4-二羟基苄基，3,4-二甲氧基苄基，4-羟基苄基，4-甲氧基苄基等；
R10为氢、—COOH、—COOCH3、—COOCH2CH3、—COOCH(CH3)2、—COO(CH2)2CH3或经卤素、羟基、C1-3烷基、C1-2烷氧基中选出1～3个取代基取代的C1-4烷基。
本发明公开了式I所示一种芳基苯并呋喃类酰胺化衍生物，其特征在于：
Figure PCTCN2017071614-appb-000004
其中：R3，或、和、R4为羟基以外的其他取代基；
其中：R9，或、和、R10为氢以外的其他取代基。
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式II所示：
Figure PCTCN2017071614-appb-000005
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式III所示：
Figure PCTCN2017071614-appb-000006
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式IV所示：
Figure PCTCN2017071614-appb-000007
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式V所示：
Figure PCTCN2017071614-appb-000008
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式VI所示：
Figure PCTCN2017071614-appb-000009
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式VII所示：
Figure PCTCN2017071614-appb-000010
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式VIII所示：
Figure PCTCN2017071614-appb-000011
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式IX所示：
Figure PCTCN2017071614-appb-000012
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式X所示：
Figure PCTCN2017071614-appb-000013
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式XI所示：
Figure PCTCN2017071614-appb-000014
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式XII所示：
Figure PCTCN2017071614-appb-000015
本发明公开了一类芳基苯并呋喃类酰胺化衍生物，结构通式如式XIII所示：
Figure PCTCN2017071614-appb-000016
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，其特征在于：
结构通式如式1所示，
Figure PCTCN2017071614-appb-000017
其中R1，R2，R3各自独立为未取代或经卤素、羟基取代的C1-4烷基，未取代或经卤素、羟基取代的C1-3烷氧基，卤素，硝基，苄基；
R4为氢，未取代或经卤素、羟基、巯基取代的C1-4烷基，3-乙基-1H-吲哚，未取代或经卤素、羟基取代的苯基，未取代或经卤素、羟基取代的苄基，未取代或经卤素、羟基取代的其他芳基(其中，卤素X＝F，Cl，Br)，如3,4-二羟基苄基，3,4-二甲氧基苄基，4-羟基苄基，4-甲氧基苄基等；
R5为氢，—COOH、—COOCH3、—COOCH2CH3、—COOCH(CH3)2、—COO(CH2)2CH3或者为氢、C1-4烷基。
本发明公开了式1所示一种芳基苯并呋喃类酰胺化衍生物，其特征在于：
结构通式如式1所示：
Figure PCTCN2017071614-appb-000018
其中R1，R2，R3各自独立为氢，羟基；R4为氢；R5为氢。
本发明公开了式1所示的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：
Figure PCTCN2017071614-appb-000019
其中R1，R2，R3各自独立为未取代或经卤素、羟基取代的C1-4烷基，未取代或经卤素、羟基取代的C1-3烷氧基，卤素，硝基，苄基；
R4为未取代或经卤素、羟基、巯基取代的C1-4烷基，3-乙基-1H-吲哚，未取代或经卤素、羟基取代的苯基，未取代或经卤素、羟基取代的苄基，未取代或经卤素、羟基取代的其他芳基(其中，卤素X＝F，Cl，Br)，如3,4-二羟基苄基，3,4-二甲氧基苄基，4-羟基苄基，4-甲氧基苄基；
R5为—COOH、—COOCH3、—COOCH2CH3、—COOCH(CH3)2、
—COO(CH2)2CH3或者为氢、C1-4烷基。
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式5a所示：
Figure PCTCN2017071614-appb-000020
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式6a所示：
Figure PCTCN2017071614-appb-000021
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式5b所示：
Figure PCTCN2017071614-appb-000022
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式6b所示：
Figure PCTCN2017071614-appb-000023
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式5c所示：
Figure PCTCN2017071614-appb-000024
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式6c所示：
Figure PCTCN2017071614-appb-000025
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式5d所示：
Figure PCTCN2017071614-appb-000026
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式6d所示：
Figure PCTCN2017071614-appb-000027
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式5e所示：
Figure PCTCN2017071614-appb-000028
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式6e所示：
Figure PCTCN2017071614-appb-000029
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式5f所示：
Figure PCTCN2017071614-appb-000030
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如式6f所示：
Figure PCTCN2017071614-appb-000031
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如7式所示：
Figure PCTCN2017071614-appb-000032
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如9a所示：
Figure PCTCN2017071614-appb-000033
本发明公开了一种芳基苯并呋喃类酰胺化衍生物，结构式如10a所示：
Figure PCTCN2017071614-appb-000034
本发明的一种芳基苯并呋喃类酰胺化衍生物的合成方法如下：
(1)将丹酚酸C与无机碱混合，超声溶解于混合溶剂中，加热不断检测反应，待反应完全后，硅胶柱色谱分离，得到中间体Tournefolic acid A(2)；
(2)将SOCl2冰浴冷却下加入有机溶剂中，反应30min后，加入丙氨酸、苯丙氨酸、半胱氨酸、酪氨酸、甲硫氨酸、色氨酸、D/L-DOPA等氨基类化合物，室温搅拌，浓缩蒸干溶剂并洗涤干燥，得系列羧基保护的氨基衍生物；
(3)将中间产物Tournefolic acid A在缩合剂作用下分别与系列氨基衍生物以摩尔比1：1.1～2混合，溶于有机溶剂中，室温搅拌，反应后处理后经硅胶柱色谱分离，得到系列的芳基苯并呋喃类酰胺化衍生物；
(4)在系列芳基苯并呋喃类酰胺化衍生物中加入无机碱，混合溶剂超声溶解，室温搅拌或加热回流，TLC不断监测反应，样品混合物经后处理，再经过硅胶柱色谱或制备型高效液相色谱分离纯化，得到系列高纯度的苯并呋喃类酰胺化衍生物。
所述的第1步中的无机碱水解可选择LiOH，NaOH，KOH等，有机溶媒为THF，MeOH，H2O或为其混合溶剂，优选为混合溶剂MeOH/H2O，且溶剂体积比优选为3：1～5：1，反应时间优选为8～12h。
所述的第1步中的分离纯化方法具体为：硅胶柱层析，用氯仿/甲醇/甲酸(10：1：0.1，v/v/v)等度洗脱得到中间体Tournefolic acid A(2)。
所述的第2步中的氨基类化合物为各种D/L构型的氨基酸及其它手性氨基衍生物，有 机溶剂为甲醇、乙醇及异丙醇等试剂，反应时间优选为18～24h。
所述的第2步中的分离纯化方法具体为：甲醇和乙醚分别多次交互洗涤样品并浓缩蒸干。
所述的第3步中的缩合剂可以为DCC/HoBt，EDCI/HoBt，HATU，HBTU，PyBOP，优选为EDCI/HoBt，有机溶剂优选为混合溶剂DMF/CH2Cl2(3：1～5：1v/v)，反应时间优选为8～12h。
所述的第3步中的分离纯化方法具体为：硅胶柱层析，氯仿/甲醇梯度洗脱得目标衍生物。
所述的第4步中的无机碱可以为LiOH，NaOH，KOH，优选为NaOH，有机溶媒为THF，MeOH，H2O或为其混合溶剂，优选为MeOH/H2O(3：1～5：1v/v)，反应时间优选为8～12h。
所述的第4步中部分芳基苯并呋喃类酰胺化衍生物具体的分离纯化方法为：硅胶柱层析分离纯化，流动相为氯仿/甲醇/甲酸梯度洗脱；或高效液相制备型色谱柱分离纯化样品混合物，其中，色谱柱(Agilent，zorbax-C18，5μm，9.4×250mm)，色谱条件优选为：流速：8mL/min，检测波长：281nm，柱温：30℃，流动相：乙腈-0.1％甲酸-水。
本制备方法中所得的化合物，均通过质谱和核磁共振等光谱学方法进行结构鉴定，并利用HPLC法进行了纯度检测。
本发明的芳基苯并呋喃类酰胺化衍生物及其药物上可接受的成盐化产物可与药用常见辅料和载体结合，制备痛风及高尿酸血症药物的组合物，从而达到预防或治疗的效果。上述药物可根据实际的需要选择合适的剂型，如片剂、注射剂、栓剂、气雾剂、纳米制剂等。
与现有技术相比较，本发明具有如下有益效果：
(1)本发明通过以丹酚酸C为底物进行了化学结构修饰，得到了系列芳基苯并呋喃类衍生物。经体外生物活性筛选及细胞活性验证，本发明的芳基苯并呋喃类酰胺化衍生物整体上均表现出较强的黄嘌呤氧化酶的抑制活性及抗氧化活性，为痛风与高尿酸血症及其并发症的防治提供了物质基础。
(2)本发明的芳基苯并呋喃类酰胺化衍生物结构新颖，对黄嘌呤氧化酶具有良好的抑制效果。与丹酚酸C相比，不同程度改善了水溶性，并在一定程度上增加了化合物的物理化学稳定性，毒副作用较低，具备一定开发应用价值。
附图说明：
图1：芳基苯并呋喃类酰胺化衍生物5a，5b，5c，6b，6c，6d，6e以及Salvianolic acid C(1)和Ve对LPS刺激巨噬细胞RAW 264.7中超氧阴离子的清除作用
图2：衍生物5b和6e和Salvianolic acid C(1)对细胞中尿酸的抑制作用
图3：活性化合物与黄嘌呤氧化酶分子对接实验结果
图3的具体解释：图3是由A部分、B部分、C部分、D部分、E部分、F部分组合而成，分别表示不同化合物与黄嘌呤氧化酶分子对接实验结果，(A)Febuxostat(B)Allopurinol(C)6b(D)6e(E)Salvianolic acid C(1)(F)5b。
具体实施方式
下面结合实施例对本发明作进一步详细描述，但本发明的实施方式不限于此。
本文中使用的测定仪器：
高分辨率质谱用美国Agilent G1969TOF/MS质谱仪；
核磁共振用瑞士Bruker AV-300或500核磁共振仪；
反应试剂均购自Sigma-Aldrich或Aladdin试剂公司，试剂均为分析纯或化学纯，氘代试剂购自美国剑桥CIL试剂公司。
实施例1
1.芳基苯并呋喃类酰胺化衍生物5a，6a的合成
(1)Tournefolic acid A
准确称取Salvianolic acid C(1)(245mg，0.498mmol)于二颈圆底瓶中，加入MeOH/H2O 20mL(5：1v/v)，超声至样品完全溶解，加入准确称取适量的无机碱(NaOH或KOH)，加热条件下不断TLC(氯仿/甲醇/甲酸8：1：1v/v/v)监测反应，直至原料完全消失。待反应完全后，放置至室温，蒸干溶剂MeOH，加入15mL蒸馏水稀释，冰浴冷却下，逐滴滴加10％HCl水溶液并不断搅拌，至pH 3～4，停止滴加。加入乙酸乙酯(20mL×3)萃取，合并有机层，饱和NaCl溶液洗涤，MgSO4干燥，减压浓缩，得反应产物粗品。经硅胶柱层析等度洗脱(氯仿/甲醇/甲酸10：1：0.1v/v/v)，得产物Tournefolic acid A(138mg，yield 89％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：深黄色粉末(TLC Rf＝0.42氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.92(d,1H,J＝15.9Hz),7.41-7.29(m,3H),7.18(s,1H),6.88(d,1H,J＝8.1Hz),6.73(d,1H,J＝8.1Hz),6.43(d,1H,J＝15.9Hz)；13C NMR(CD3OD,75MHz)δ171.3,159.3,148.0,146.7,145.7,144.6,144.3,132.5,126.1,123.4,119.7,118.7,116.7,116.0,113.3,111.7,99.2.HR-MS(ESI)m/z:found 311.0563[M-H]-,calcd.for C17H12O6 311.0561.
据以上波谱信息，可鉴定化合物Tournefolic acid A化学式为C17H12O6，其结构如下式。
Figure PCTCN2017071614-appb-000035
(2)Methyl(S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride(4a)
准确量取无水甲醇5mL于圆底烧瓶中，冰浴冷却15min，缓慢逐滴加入SOCl2(493μL，6.780mmol)，0℃搅拌30min后，加入准确称取的L-DOPA(800mg，4.520mmol)，放置至室温，搅拌反应24h。待反应完全后，蒸干溶剂和反应剩余的SOCl2，用MeOH和Et2O分别洗涤样品三次，蒸干，得反应产物L-多巴甲酯盐酸盐(1.03g，yield 93％)。
对步骤(2)中得到的反应产物进行结构解析。
理化性质：白色粉末(TLC Rf＝0.64氯仿/甲醇10：1)。
波谱信息：1H NMR(D2O,300MHz)δ6.78(d,1H,J＝8.2Hz),6.78(d,1H,J＝1.8Hz), 6.68(dd,1H,J＝1.8,8.1Hz),4.35(t,1H,J＝6.6Hz),3.82(s,3H),3.18(dd,1H,J＝5.9,14.6Hz),3.06(dd,1H,J＝7.7,14.6Hz).13C NMR(D2O,75MHz)δ170.6,144.8,144.2,126.6,122.3,117.4,117.1,54.8,54.0,35.4.ESI-MS m/z:212.3[M+H]+.
据以上波谱信息，可鉴定化合物4a化学式为C10H14ClNO4，结构如下式。
Figure PCTCN2017071614-appb-000036
(3)Methyl(S,E)-3-(3,4-dihydroxyphenyl)-2-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acrylamido)propanoate(5a)
准确称取Tournefolic acid A(36mg，0.115mmol)于圆底烧瓶中，加入DMF/CH2Cl2 4mL(4：1v/v)，超声溶解，冰浴冷却，加入准确称量的EDCI(33.1mg，0.173mmol)，HoBt(23.3mg，0.173mmol)，冰浴冷却10min，并不断搅拌。加入准确称取的L-多巴甲酯盐酸盐(34.1mg，0.138mmol)，Et3N(34.8mg，0.345mmol)，冰浴中反应30min后置室温搅拌，TLC(氯仿/甲醇/甲酸8：1：1v/v/v)不断监测反应，直至原料基本消失。待停止反应后，蒸干溶剂，加15mL蒸馏水稀释样品，冰浴冷却，滴加几滴10％HCl水溶液，充分震荡。用乙酸乙酯(15mL×4)萃取，合并有机层，减压浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经硅胶柱分离纯化样品(氯仿/甲醇15：1→10：1v/v)，得产物5a(29.6mg，yield 51％)。
对步骤(3)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.61氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.74(d,1H,J＝15.6Hz),7.39(s,1H),7.34(d,1H,J＝8.7Hz),7.28(s,1H),6.87(d,1H,J＝8.1Hz),6.74-6.63(m,4H),6.55(d,1H,J＝7.2Hz),4.76-4.72(m,1H),3.72(s,3H),3.11-3.00(dd,1H,J＝13.5,6.0Hz),2.99-2.89(dd,1H,J＝13.5,7.5Hz)；13C NMR(CD3OD,75MHz)δ173.8,169.3,159.1,147.9,146.8,146.3,145.3,145.3,144.5,140.7,132.0,129.5,126.0,123.5,121.7,120.2,118.6,118.4,117.3,116.7,116,4,113.4,111.6,99.5,55.8,52.6,38.2.HR-MS(ESI)m/z:found 504.1302[M-H]-,calcd.for C27H23NO9504.1300.
据以上波谱信息，可鉴定化合物5a化学式为C27H23NO9，其结构式如下。
Figure PCTCN2017071614-appb-000037
(4)(S,E)-3-(3,4-dihydroxyphenyl)-2-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acrylamido)propanoic acid(6a)
准确称取化合物5a(18mg，0.036mmol)于二颈烧瓶中，加入MeOH/H2O 5mL (4：1v/v)溶解，加入NaOH(17.4mg，0.44mmol)，加热回流，TLC(氯仿/甲醇/甲酸8：1：1)监测反应至反应完全。待反应停止，冰浴冷却反应混合液，滴加10％HCl水溶液并不断搅拌，至pH 3～4，浓缩蒸干，加入蒸馏水10mL，用乙酸乙酯(10mL×4)萃取，合并有机层，减压浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经制备型液相色谱(Agilent zorbax-C18，5μm，20×250mm)分离，流速8mL/min，检测波长281nm，流动相：乙腈-0.1％甲酸-水，得目标产物6a(11.6mg，yield 66％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.48氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(DMSO-d6,300MHz)δ8.15(br s,4H),7.54(d,1H,J＝15.9Hz),7.43(s,1H),7.35(s,1H),7.28(d,1H,J＝8.1Hz),7.23(d,1H,J＝8.4Hz),6.88(d,1H,J＝8.1Hz),6.79-6.72(q,2H,J＝15.9Hz),6.65-6.60(m,2H),6.50(d,1H,J＝8.1Hz),4.50(m,1H),3.00-2.91(d,1H,J＝14.1,5.1Hz),2.84-2.74(d,1H,J＝13.5,8.7Hz)；13C NMR(DMSO-d6,75MHz)δ173.3,165.6,156.9,146.9,145.7,145.0,143.9,143.8,142.7,137.8,129.6,128.3,125.7,121.1,119.9,119.2,118.6,117.0,116.5,116.1,115.4,112.4,110.6,99.0,53.9,36.7.HR-MS(ESI)m/z:found 492.1296[M+H]+,calcd.for C26H21NO9 492.1289.
据以上波谱信息，可鉴定化合物6a化学式为C26H21NO9，其结构式如下。
Figure PCTCN2017071614-appb-000038
实施例2
2.芳基苯并呋喃类酰胺化衍生物5b，6b的合成
(1)-(2)同实施例1中制备中间体的方法。
(3)Methyl(R,E)-3-(3,4-dihydroxyphenyl)-2-(3-(2-(3,4-dihydroxyphenyl)
-7-hydroxybenzofuran-4-yl)acrylamido)propanoate(5b)
准确称取Tournefolic acid A(32mg，0.103mmol)于圆底烧瓶中，加入DMF/CH2Cl23.5mL(5：1v/v)，超声至样品溶解，冰浴冷却，加入准确称量的EDCI(31.6mg，0.165mmol)，HoBt(22.3mg，0.165mmol)，冰浴冷却10min，并不断搅拌。加入准确称取的D-多巴甲酯盐酸盐(30.5mg，0.124mmol)，Et3N(31.2mg，0.309mmol)，冰浴中反应30min后置室温搅拌，TLC(氯仿/甲醇/甲酸8：1：1)监测反应至原料基本消失。待停止反应后，蒸干溶剂，加15mL蒸馏水稀释样品，冰浴冷却，滴加几滴10％HCl水溶液，充分震荡。用乙酸乙酯(15mL×4)萃取，合并有机层，减压浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经硅胶柱纯化(氯仿/甲醇15：1→10：1v/v)得产物5b(30mg，yield 58％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.6氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.74(d,1H,J＝15.6Hz),7.39(s,1H),7.37-7.32 (d,1H,J＝8.4Hz),7.29-7.24(m,2H),6.87(d,1H,J＝8.1Hz),6.73-6.65(m,4H),6.55(d,1H,J＝8.4Hz),5.08(m,1H),4.06(s,3H),4.00(s,3H),3.94(s,3H),3.77(s,3H),3.23(m,1H),2.93(m,1H)；13C NMR(CD3OD,75MHz)δ172.4,165.6,157.0,146.9,145.7,145.0,144.5,143.9,142.7,138.0,129.7,127.8,125.7,121.1,119.8,118.7,118.4,117.0,116.4,116.1,115,4,112.3,110.6,96.9,54.0,51.8,36.7.HR-MS(ESI)m/z:found 504.1311[M-H]-,calcd.for C27H23NO9504.1300.
据以上波谱信息，可鉴定化合物5b化学式为C27H23NO9，其结构式如下。
Figure PCTCN2017071614-appb-000039
(4)(R,E)-3-(3,4-dihydroxyphenyl)-2-(3-(2-(3,4-dihydroxyphenyl)
-7-hydroxybenzofuran-4-yl)acrylamido)propanoic acid(6b)
准确称取化合物5b(18mg，0.036mmol)于二颈烧瓶中，加入MeOH/H2O 5mL(3：1v/v)溶解，准确称取NaOH(17.1mg，0.428mmol)加入上述溶液中，加热回流，TLC(氯仿/甲醇/甲酸8：1：1)监测反应，直至反应完全。待停止反应，冰冷却反应液，滴加10％HCl溶液并搅拌，至pH 3～4，蒸干，加入蒸馏水10mL，用乙酸乙酯(10mL×4)萃取，合并有机层，浓缩，饱和食盐水洗涤，MgSO4干燥，样品混合物经制备型液相色谱(Agilent zorbax-C18，5μm，20×250mm)分离，流速8mL/min，检测波长281nm，流动相：乙腈-0.1％甲酸-水，得产物6b(9.5mg，yield 54％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄色粉末(TLC Rf＝0.46氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(DMSO-d6,300MHz)δ8.21(br s,4H),7.53(d,1H,J＝15.9Hz),7.43(s,1H),7.35(s,1H),7.23(d,1H,J＝8.1Hz),7.22(d,1H,J＝8.1Hz),6.87(d,1H,J＝8.4Hz),6.80-6.70(q,2H,J＝15.9Hz),6.65-6.59(m,2H),6.43(d,1H,J＝8.4Hz),4.50(m,1H),2.98-2.91(d,1H,J＝14.2,4.9Hz),2.81-2.73(d,1H,J＝14.2,8.3Hz)；13C NMR(DMSO-d6,75MHz)δ173.6,165.4,156.9,146.9,145.7,144.9,143.8,143.8,142.7,137.6,129.5,128.5,125.7,121.0,119.9,119.3,118.6,116.9,116.6,116.1,115.3,112.4,110.5,99.0,54.2,36.8.HR-MS(ESI)m/z:found 492.1293[M+H]+,calcd.for C26H21NO9 492.1289.
根据以上高分辨质谱及核磁谱信息，可鉴定化合物6b的化学式为C26H21NO9，其结构式如下。
Figure PCTCN2017071614-appb-000040
实施例3
3.芳基苯并呋喃类酰胺化衍生物5c，6c的合成
(1)-(2)同实施例1中制备中间体的方法。
(3)Methyl(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl)-L-alaninate(5c)
准确称取Tournefolic acid A(35mg，0.112mmol)于圆底烧瓶中，加入DMF/CH2Cl2 4mL(3：1v/v)，超声至样品溶解，冰浴冷却至0℃，加入EDCI(32.3mg，0.168mmol)，HoBt(22.6mg，0.168mmol)，冰浴冷却10min，并不断搅拌。加入准确称取的L-丙氨酸甲酯盐酸盐(18.7mg，0.134mmol)，Et3N(33.9mg，0.336mmol)，冰浴中反应30min后放置至室温搅拌，TLC(氯仿/甲醇/甲酸8：1：1)不断监测反应，直至原料基本消失。待停止反应后，蒸干溶剂，加15mL蒸馏水稀释样品，冰浴冷却，滴加几滴10％HCl水溶液，充分震荡。用乙酸乙酯(15mL×4)萃取，合并有机层，减压浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经硅胶柱(氯仿/甲醇20：1→8：1v/v)纯化，得产物5c(28.4mg，yield 64％)。
对步骤(3)中得到的反应产物进行结构解析。
理化性质：淡绿色粉末(TLC Rf＝0.65氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.77(d,1H,J＝15.9Hz),7.39(s,1H),7.35(d,1H,J＝8.1Hz),7.32-7.25(m,2H),6.87(d,1H,J＝8.1Hz),6.75-6.70(d,1H,J＝8.4Hz),6.70-6.63(d,1H,J＝15.9Hz),4.62-4.53(dd,1H,J＝14.7,7.5Hz),3.75(s,3H),1.46(d,3H,J＝7.2Hz)；13C NMR(CD3OD,75MHz)δ174.9,169.2,159.1,148.0,146.8,145.4,144.5,140.7,132.1,125.9,123.5,120.2,118.6,118.4,116.7,113.4,111.7,99.5,52.8,49.7,17.7.HR-MS(ESI)m/z:found 396.1089[M-H]-,calcd.for C21H19NO7 396.1089.
根据以上高分辨质谱及核磁谱信息，可鉴定化合物5c的化学式为C21H19NO7，其结构式如下。
Figure PCTCN2017071614-appb-000041
(4)(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl) -L-alanine(6c)
准确称取化合物5c(19mg，0.048mmol)于二颈烧瓶中，加入MeOH/H2O 5mL(3：1v/v)溶解，加入NaOH(19.1mg，0.478mmol)，加热回流，并TLC(氯仿/甲醇/甲酸8：1：1)监测反应，至反应完全。待停止反应后，冰浴冷却反应混合液，滴加10％HCl水溶液并不断搅拌，至pH 3～4，浓缩蒸干MeOH，加入蒸馏水10mL，用乙酸乙酯(10mL×4)萃取，合并有机层，浓缩，饱和食盐水洗涤，MgSO4干燥，样品混合物经制备型液相色谱(Agilent zorbax-C18，5μm，20×250mm)分离，流速8mL/min，检测波长281nm，流动相：乙腈-0.1％甲酸-水，得衍生物6c(13.1mg，yield 71％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.48氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.53(d,1H,J＝15.6Hz),7.44(s,1H),7.36(s,1H),7.32-7.20(m,2H),6.83(d,1H,J＝7.8Hz),6.82(t,2H,J＝15.6Hz),4.38(m,1H),1.35(d,3H,J＝6.6Hz)；13C NMR(CD3OD,75MHz)δ174.5,165.5,157.0,147.0,145.8,143.9,142.8,137.8,129.6,125.8,121.1,119.3,118.7,117.0,116.2,112.4,110.6,99.0,48.7,17.8.HR-MS(ESI)m/z:found 382.0941[M-H]-,calcd.for C20H17NO7 382.0932.
据以上波谱信息，可鉴定化合物6c化学式为C20H17NO7，其结构式如下。
Figure PCTCN2017071614-appb-000042
实施例4
4.芳基苯并呋喃类酰胺化衍生物5d，6d的合成
(1)-(2)同实施例1中制备中间体的方法。
(3)Methyl(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl)-L-phenylalaninate(5d)
准确称取Tournefolic acid A(35mg，0.112mmol)于25mL圆底烧瓶中，加入DMF/CH2Cl2 3mL(4：1v/v)，超声至样品溶解，冰浴冷却，加入EDCI(30.2mg，0.157mmol)，HoBt(21.2mg，0.157mmol)，冰浴冷却10min，并不断搅拌。加入L-苯丙氨酸甲酯盐酸盐(28.9mg，0.134mmol)，Et3N(33.9mg，0.336mmol)，冰浴中反应30min后放置至室温搅拌，TLC(氯仿/甲醇/甲酸8：1：1)不断监测反应，直至原料基本消失。待停止反应后，蒸干溶剂，加15mL蒸馏水稀释样品，冰浴冷却下滴加几滴10％HCl水溶液，充分震荡。用乙酸乙酯(15mL×3)萃取，合并有机层，浓缩，饱和食盐水洗涤，MgSO4干燥，混合物经硅胶柱(氯仿/甲醇20：1→10：1v/v)纯化，得反应产物5d(24.3mg，yield 46％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.69氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.73(d,1H,J＝15.9Hz),7.39(s,1H),7.34(d, 1H,J＝9.0Hz),7.31-7.18(m,7H),6.87(d,1H,J＝8.1Hz),,6.71(d,1H,J＝8.4Hz),6.65(d,1H,J＝15.9Hz),4.95-4.86(m,1H),3.72(s,3H),3.27-3.17(dd,1H,J＝13.5,5.7Hz),3.12-3.01(dd,1H,J＝13.5,8.4Hz)；13C NMR(CD3OD,75MHz)δ173.7,169.3,159.1,148.0,146.8,145.3,140.8,138.2,132.1,130.2,130.2,129.5,129.5,127.9,126.0,123.5,121.8,120.2,118.6,118.3,116.7,113.4,111.6,99.5,55.6,52.7,38.7.HR-MS(ESI)m/z:found 472.1414[M-H]-,calcd.for C27H23NO7 472.1402.
据以上波谱信息，可鉴定化合物5d化学式为C27H23NO7，其结构式如下。
Figure PCTCN2017071614-appb-000043
(4)(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl)
-L-phenylalanine(6d)
准确称取化合物5d(16mg，0.034mmol)于二颈烧瓶中，加入MeOH/H2O 5mL(4：1v/v)溶解，加入NaOH(14.9mg，0.372mmol)，加热回流，并TLC(氯仿/甲醇/甲酸8：1：1)监测反应，至反应完全。待停止反应后，冰浴冷却反应液，滴加10％HCl水溶液，并不断搅拌至pH 3～4，浓缩蒸干，加入蒸馏水10mL，乙酸乙酯(10mL×4)萃取，合并有机层，浓缩，饱和食盐水洗涤，MgSO4干燥，样品混合物经制备型液相色谱(Agilent zorbax-C18，5μm，20×250mm)分离，流速8mL/min，检测波长281nm，流动相：乙腈-0.1％甲酸-水，得反应产物6d(8.1mg，yield 52％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.53氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(DMSO-d6,300MHz)δ8.20(br s,2H),7.54(d,1H,J＝15.9Hz),7.42(s,1H),7.35(s,1H),7.30-7.20(m,7H),6.88(d,1H,J＝7.8Hz),6.79-6.69(t,2H,J＝15.9Hz),4.63(m,1H),3.16-3.08(m,1H),3.01-2.91(m,1H)；13C NMR(DMSO-d6,75MHz)δ173.2,165.5,156.9,146.9,145.7,143.8,142.7,137.73,137.68,129.6,129.1,129.1,128.2,128.2,126.4,125.6,121.0,119.0,118.5,116.9,116.1,112.4,110.5,96.9,53.7,37.2.HR-MS(ESI)m/z:found 458.1252[M-H]-,calcd.for C26H21NO7 458.1245.
据以上波谱信息，可鉴定化合物6d化学式为C26H21NO7，其结构式如下。
Figure PCTCN2017071614-appb-000044
实施例5
5.芳基苯并呋喃类酰胺化衍生物5e，6e的合成
(1)-(2)同实施例1中制备中间体的方法。
(3)Methyl(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl)-L-tyrosinate(5e)
准确称取Tournefolic acid A(33mg，0.106mmol)于圆底烧瓶中，加入DMF/CH2Cl2 4mL(3：1v/v)，超声至样品完全溶解，冰浴冷却，加入EDCI(30.4mg，0.158mmol)，HoBt(21.5mg，0.159mmol)，冰浴冷却10min，并不断搅拌。加入L-酪氨酸甲酯盐酸盐(29.4mg，0.127mmol)，Et3N(32.1mg，0.318mmol)，冰浴中反应30min后至室温搅拌，TLC(氯仿/甲醇/甲酸8：1：1)不断监测反应，直至原料基本消失。待停止反应后，蒸干溶剂，加15mL蒸馏水稀释样品，冰浴冷却，滴加10％HCl水溶液，充分震荡。用乙酸乙酯(15mL×4)萃取，合并有机层，减压浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经硅胶柱(氯仿/甲醇15：1→10：1v/v)分离纯化，得衍生物5e(28.0mg，yield 54％)。
对步骤(3)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.64氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.73(d,1H,J＝16.2Hz),7.39(s,1H),7.37-7.32(d,1H,J＝8.1Hz),7.26(s,1H),7.06(d,2H,J＝8.4Hz),6.90-6.82(m,2H,J＝7.5Hz),6.74-6.64(m,4H),4.69-4.58(m,1H),3.72(s,3H),3.16-3.07(dd,1H,J＝13.5,6.0Hz),3.02-2.94(dd,1H,J＝13.5,8.1Hz)；13C NMR(CD3OD,75MHz)δ174.5,169.2,159.3,155.7,147.3,146.6,145.2,143.9,141.7,130.2,130.2,129.2,124.3,123.0,122.0,121.6,120.2,119.8,116.9,115.8,115.8,114.2,108.4,99.8,56.2,52.0,38.2.HR-MS(ESI)m/z:found 488.1353[M-H]-,calcd.for C27H23NO8 488.1351.
据以上波谱信息，可鉴定化合物5e化学式为C27H23NO8，其结构式如下。
Figure PCTCN2017071614-appb-000045
(4)(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl)-L-tyrosine(6e)
准确称取化合物6e(19mg，0.039mmol)于二颈烧瓶中，加入MeOH/H2O 5mL(4：1v/v)溶解，加入NaOH(14.0mg，0,350mmol)，加热回流反应，并TLC(氯仿/甲醇/甲酸8：1：1)监测反应至反应完全。停止反应后，冰浴冷却反应液，滴加10％HCl水溶液并不断搅拌，至pH 3～4，浓缩蒸干，加入蒸馏水10mL，用乙酸乙酯(10mL×4)萃取，合并有机层，减压浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经制备型液相色谱(Agilent zorbax，SB-C18，5μm，20×250mm)分离，流速8mL/min，检测波长281nm，流动相：乙腈-0.1％甲酸-水，得反应产物6e(11.8mg，yield 62％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.5氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(DMSO-d6,300MHz)δ7.95(br s,1H),7.50(d,1H,J＝16.2Hz),7.47(s,1H),7.36(s,1H),7.28(d,1H,J＝8.1Hz),7.20(d,1H,J＝8.1Hz),7.06-6.98(m,2H),6.87(d,1H,J＝8.1Hz),6.79(d,1H,J＝16.2Hz),6.74(d,1H,J＝8.1Hz),6.65-6.59(m,2H),4.66(m,1H),3.09-3.01(q,1H),2.89-2.80(q,1H)；13C NMR(DMSO-d6,75MHz)δ174.3,165.2,156.9,155.7,147.0,145.8,143.8,142.7,137.3,130.1,130.1,129.4,128.4,125.8,121.0,119.6,118.6,116.9,116.2,114.8,114.8,112.5,112.5,99.1,54.8,36.8.HR-MS(ESI)m/z:found 474.1201[M-H]-,calcd.for C26H21NO8 474.1194.
据以上波谱信息，可鉴定化合物6e化学式为C26H21NO8，其结构式如下。
Figure PCTCN2017071614-appb-000046
实施例6
6.芳基苯并呋喃类酰胺化衍生物5f，6f的合成
(1)-(2)同实施例1中制备中间体的方法。
(3)Methyl(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl)-L-tryptophanate(5f)
准确称取Tournefolic acid A(41mg，0.131mmol)于圆底烧瓶中，加入DMF/CH2Cl2 5mL(5：1v/v)，超声至样品完全溶解，冰浴冷却至0℃，加入准确称量的EDCI(37.8mg，0.197mmol)，HoBt(26.5mg，0.196mmol)，冰浴冷却10min，并不断搅拌。加入准确称取的L-色氨酸甲酯盐酸盐(39.9mg，0.157mmol)，Et3N(39.7mg，0.393mmol)，冰浴中反应30min后放置至室温搅拌，TLC(氯仿/甲醇/甲酸8：1：1)不断监测反应，直至原料基本消失。待停止反应后，蒸干溶剂，加15mL蒸馏水稀释样品，滴加10％HCl水溶液，冰浴冷却下充分震荡。用乙酸乙酯(15mL×4)萃取，合并有机层，减压浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经硅胶柱分离纯化(氯仿/甲醇20：1→12：1v/v)，得衍生物5f(28.2mg，yield 42％)。
对步骤(3)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.71氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.74(d,1H,J＝15.6Hz),7.56(d,1H,J＝7.5Hz),7.40-7.29(m,3H),7.28-7.20(m,2H),7.11(s,1H),7.10-7.05(d,1H,J＝7.5Hz),7.04-6.98(t,1H,J＝7.5,7.2Hz),6.86(d,1H,J＝6.6Hz),6.75-6.69(d,1H,J＝6.0Hz),6.69-6.62(d,1H,J＝15.6Hz),4.95-4.89(m,1H),3.69(s,3H),3.43-3.35(m,1H),3.29-3.20(m,1H)；13C NMR(CD3OD,75MHz)δ174.2,169.4,159.1,148.2,147.0,145.9,144.7,140.8,138.9,138.1,132.0,128.8,126.1,124.4,123.5,122.5,120.0,119.9,119.2,118.6,118.3,117.3,116.7,113,4,112.3,111.8,99.5,55.2,52.7,28.8.HR-MS(ESI)m/z:found 511.1508[M-H]-,calcd.for C20H17NO7 511.1511.
据以上波谱信息，可鉴定化合物5f化学式为C29H24N2O7，其结构式如下。
Figure PCTCN2017071614-appb-000047
(4)(E)-(3-(2-(3,4-dihydroxyphenyl)-7-hydroxybenzofuran-4-yl)acryloyl)-D-tryptophan(6f)
准确称取化合物5f(20mg，0.039mmol)于二颈烧瓶中，加入MeOH/H2O 5mL(4：1v/v)溶解，加入NaOH(18.8mg，0.469mmol)，加热回流反应，并TLC(氯仿/甲醇/甲酸8：1：1)监测反应，至反应完全。待反应停止，冰浴冷却反应混合液，滴加10％HCl液并不断搅拌，至pH 3～4，浓缩蒸干，加入蒸馏水10mL，用乙酸乙酯(10mL×4)萃取，合并有机层，浓缩，饱和食盐水洗涤，MgSO4干燥。样品混合物经制备型液相色谱(Agilent zorbax-C18，5μm，20×250mm)分离，流速8mL/min，检测波长281nm，流动相：乙腈-0.1％甲酸-水，得反应产物6f(10.7mg，yield 55％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.58氯仿/甲醇/甲酸8：1：1)。
波谱信息：1H NMR(DMSO-d6,300MHz)δ10.85(s,1H),8.17(s,1H),7.62-7.56(t,2H,J＝16.2Hz),7.44(s,1H),7.37(s,1H),7.34(d,1H,J＝8.4Hz),7.29(d,1H,J＝8.7Hz),7.24(d,1H,J＝8.1Hz),7.18(s,1H),7.09-7.03(t,1H,J＝7.5Hz),7.01-6.96(t,1H,J＝7.5Hz),6.89(d,1H,J＝8.1Hz),6.80(d,1H,J＝16.2Hz),6.75(d,1H,J＝9.3Hz),4.72(dd,1H,J＝12.8,7.4Hz),3.69(dd,1H,J＝14.4,4.8Hz),3.29-3.20(dd,1H,J＝14.4,7.8Hz)；13C NMR(DMSO-d6,75MHz)δ173.7,165.7,157.0,147.0,145.8,143.9,142.8,137.9,136.2,129.6,127.4,125.9,123.6,121.1,121.0,119.3,118.7,118.5,118.4,117.0,116.2,112.4,111.5,110.6,110.0,99.1,53.2,27.6.HR-MS(ESI)m/z:found 497.1368[M-H]-,calcd.for C28H22N2O7 497.1354.
据以上波谱信息，可鉴定化合物6f化学式为C28H22N2O7，其结构式如下。
Figure PCTCN2017071614-appb-000048
实施例7
7.芳基苯并呋喃类酰胺化衍生物7的合成
(2R)-3-(3,4-dimethoxyphenyl)-1-methoxy-1-oxopropan-2-yl(2E)-3-[2-(3,4- dimethoxyphenyl)-7-methoxy-1-benzofuran-4-yl]prop-2-enoate(7)
准确称取Salvianolic acid C(206mg，0.419mmol)于圆底二颈烧瓶中，加15mL Me2CO超声溶解，准确称(量)取K2CO3(1.16g，8.374mmol)，Me2SO4(714.5mL，7.542mmol)加入salvianolic acid C样品溶液中，回流反应，并TLC(环己烷/乙酸乙酯1：1)监测，直至反应原料基本消失。待反应终止，蒸干溶剂，饱和NH4Cl水溶液(20mL)稀释，用乙酸乙酯(20mL×4)萃取，合并有机层，饱和食盐水洗涤，MgSO4干燥，减压浓缩，得混合物样品。用硅胶柱梯度洗脱(环己烷/乙酸乙酯10：1，5：1v/v)，得反应产物7(431mg，yield 92％)。
对步骤(1)中得到的反应产物进行结构分析。
理化性质：黄色粉末(TLC Rf＝0.62环己烷/乙酸乙酯1：1)。
波谱信息：1H NMR(CDCl3,300MHz)δ7.96(d,1H,J＝15.9Hz),7.53(dd,1H,J＝8.4,2.1Hz),7.39(m,2H),7.17(s,1H),6.97(d,1H,J＝8.4Hz),6.82(m,4H),6.50(d,1H,J＝15.9Hz),5.41(q,1H,J＝13.2,7.5,5.4Hz),4.09(s,3H),4.03(s,3H),3.96(s,3H),3.88(s,3H),3.86(s,3H),3.78(s,3H),3.21(dd,2H,J＝5.1,2.1Hz)；13C NMR(CDCl3,75MHz)δ170.5,166.7,157.6,150.1,149.2,148.8,148.8,146.9,144.1,143.7,130.7,128.4,125.8,122.7,121.4,121.4,118.5,114.9,112.5,111.3,111.2,108.2,106.6,99.2,73.1,56.2,56.2,56.0,55.8,55.8,52.4,37.2.HR-MS(ESI)m/z:found 577.2068[M+H]+,calcd.for C32H32O10 577.2079.
根据以上波谱信息，可鉴定化合物7化学式为C32H32O10，其结构如下式。
Figure PCTCN2017071614-appb-000049
实施例8
8.芳基苯并呋喃类酰胺化衍生物9a的合成
Methyl(S,E)-3-(3,4-dihydroxyphenyl)-2-(3-(2-(3,4-dimethoxyphenyl)
-7-methoxybenzofuran-4-yl)acrylamido)propanoate(9a)
准确称取反应中间体于圆底烧瓶中，加入DMF/CH2Cl2 5mL(v/v 2：5)，超声至样品溶解，冰浴冷却至0℃，加入EDCI(27.4mg，0.143mmol)，HoBt(19.3mg，0.143mmol)，冰浴冷却10min并不断搅拌。加入L-丙氨酸甲酯盐酸盐(30.3mg，0.169mmol)，Et3N(36.1mg，0.357mmol)，冰水浴中反应30min后放置至室温，TLC(环己烷/乙酸乙酯5：2v/v)监测反应，至原料基本消失。待反应完全，蒸干溶剂，加入蒸馏水15mL稀释样品，滴加几滴10％HCl水溶液，充分震荡。用乙酸乙酯(15mL×3)萃取，得有机层，浓缩，并依次用饱和NaHCO3水溶液和饱和食盐水洗涤，MgSO4干燥，得样品混合物。经硅胶柱层析(氯仿/甲醇80：1)等度洗脱，得到反应产物9a(37.7mg，yield 52％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.57环己烷/乙酸乙酯5：2)。
波谱信息：1H NMR(CD3OD,300MHz)δ7.71(d,1H,J＝15.6Hz),7.46(d,1H,J＝8.7Hz),7.42(s,1H),7.37-7.26(d,2H,J＝8.4Hz),7.01(d,1H,J＝7.8Hz),6.83(d,1H,J＝8.1Hz),6.73-6.65(t,3H,J＝15.6Hz),6.56(d,1H,J＝8.1Hz),4.77-4.60(dd,1H,J＝14.4,7.8Hz),4.02(s,3H),3.92(s,3H),3.87(s,3H),3.72(s,3H),3.13-3.01(dd,1H,J＝13.2,5.4Hz),2.99-2.91(dd,1H,J＝13.2,7.8Hz)；13C NMR(CD3OD,75MHz)δ173.8,169.0,158.5,151.5,150.8,147.8,146.3,145.4,140.5,131.4,129.5,126.2,124.4,122.7,121.7,121.6,119.54,119.49,117.3,116.4,113.2,109.8,107.9,100.3,56.71,56.67,56.5,55.8,52.7,38.2.HR-MS(ESI)m/z:found546.1777[M-H]-,calcd.for C30H29NO9 546.1770.
根据以上波谱信息，可鉴定化合物9a化学式为C30H29NO9，其结构如下。
Figure PCTCN2017071614-appb-000050
实施例9
9.芳基苯并呋喃类酰胺化衍生物10a的合成
(S,E)-3-(3,4-dihydroxyphenyl)-2-(3-(2-(3,4-dimethoxyphenyl)-7-methoxybenzofuran-4-yl)acrylamido)propanoic acid(10a)
准确称取化合物9a(20mg，0.039mmol)于二颈烧瓶中，加入MeOH/H2O 5mL(4：1v/v)溶解，加入NaOH(18.8mg，0.469mmol)，加热回流，并TLC(环己烷/乙酸乙酯/甲酸2：1：0.1％v/v/v)监测反应至反应完全。待反应停止，冰浴冷却反应液，滴加10％HCl液并不断搅拌，至pH 3～4，浓缩蒸干，加入蒸馏水10mL，乙酸乙酯(10mL×4)萃取，合并有机层，浓缩，饱和食盐水洗涤，MgSO4干燥。样品经制备型液相色谱(Agilent zorbax-C18，5μm，20×250mm)分离，流速8mL/min，检测波长281nm，流动相：乙腈-0.1％甲酸-水，得反应产物10a(10.7mg，yield 55％)。
对步骤(1)中得到的反应产物进行结构解析。
理化性质：黄绿色粉末(TLC Rf＝0.36环己烷/乙酸乙酯/甲酸4：1：0.1％)。
波谱信息：1H NMR(DMSO-d6,300MHz)δ8.20-8.09(t,1H,J＝7.8Hz),7.68(s,1H),7.62(d,1H,J＝15.9Hz),7.55-7.45(m,2H),7.41(d,1H,J＝8.7Hz),7.11(d,1H,J＝8.1Hz),6.98(d,1H,J＝8.4Hz),6.82(d,1H,J＝15.9Hz),6.67-6.58(m,2H),6.49(d,1H,J＝7.8Hz),4.58-4.46(dd,1H,J＝13.6,7.4Hz),4.01(s,3H),3.89(s,3H),3.83(s,3H),3.00-2.92(dd,1H,J＝14.1,4.8Hz),2.83-2.75(dd,1H,J＝13.6,7.4Hz)；13C NMR(DMSO-d6,75MHz)δ173.4,165.2,156.5,149.9,149.2,145.7,144.9,143.9,143.0,136.8,129.5,128.4,124.6,122.2,120.6,120.3,119.9,117.9,116.6,115.3,112.1,108.4,107.2,100.0,56.0,55.8,55.7,54.2,36.7.HR-MS(ESI)m/z:found 532.1620[M-H]-,calcd.for C29H27NO9 532.1613.
根据以上波谱信息，可鉴定化合物10a的化学式为C29H27NO9，其结构如下式。
Figure PCTCN2017071614-appb-000051
实施例10
10.芳基苯并呋喃类酰胺化衍生物9b的合成
Methyl(R,E)-3-(3,4-dihydroxyphenyl)-2-(3-(2-(3,4-dimethoxyphenyl)-7-methoxybenzofuran-4-yl)acrylamido)propanoate（9b）
准确称取反应中间体于圆底烧瓶中，加入DMF/CH2Cl2 5mL（v/v 2：5），超声至样品溶解，冰浴冷却至0℃，加入EDCI（31.4mg，0.152mmol），HoBt（26.3mg，0.167mmol），冰浴冷却10min并不断搅拌。加入D-丙氨酸甲酯盐酸盐（37mg，0.181mmol），Et3N（39mg，0.371mmol），N2保护，冰浴中反应30min后放置至室温，TLC（环己烷/乙酸乙酯5：2v/v）不断监测反应，直至原料基本消失。待反应完全后，蒸干溶剂，加入蒸馏水15mL稀释样品，滴加几滴10%HCl水溶液，充分震荡。用乙酸乙酯（15mL×3）萃取，得有机层，浓缩，并依次用饱和NaHCO3水溶液和饱和食盐水洗涤，MgSO4干燥，得样品混合物。经硅胶柱层析（环己烷/乙酸乙酯12：1）等度洗脱，得到反应产物9b（45.7mg，yield 66%）。
对步骤（1）中得到的反应产物进行结构解析。
理化性质：黄绿色粉末（TLC Rf=0.56环己烷/乙酸乙酯5：2）。
波谱信息：1H NMR(CDCl3,300MHz)δ7.61(d,1H,J=15.3Hz),7.46-7.39(m,1H),7.37-7.28(m,2H),7.12(d,1H,J=8.1Hz),6.93-6.89(m,2H),6.65-6.60(d,2H,J=8.1Hz),6.53-6.48(d,1H,J=8.4Hz),6.28(d,1H,J=15.3Hz),5.02-4.90(dd,1H,J=13.8,4.8Hz),3.97(s,3H),3.91(s,3H),3.74(s,3H),3.49(s,3H),3.24-3.11(dd,1H,J=14.4,5.4Hz),3.09-2.95(dd,1H,J=14.4,4.8Hz);13C NMR(CDCl3,75MHz)δ172.5,167.2,157.3,150.0,149.3,146.5,144.4,143.9,140.1,136.0,130.9,127.9,124.1,122.9,121.2,119.8,118.6,117.3,116.3,115.4,111.4,108.4,106.6,99.0,56.3,56.2,56.1,54.0,52.6,37.3.HR-MS(ESI)m/z:found 546.1777[M-H]-,calcd.for C30H29NO9 546.1770.
根据以上波谱信息，可鉴定化合物9b化学式为C30H29NO9，其结构如下。
Figure PCTCN2017071614-appb-000052
实施例11
芳基苯并呋喃类酰胺化衍生物及反应试剂(氨基衍生物)的黄嘌呤氧化酶抑制活性测定
11.1试剂与标准溶液的配制
(1)75mM磷酸缓冲液(PB，pH 7.4)：含KH2PO4 0.0956g，K2HPO4 0.6946g，EDTA 1.862mg，用纯水稀释至50mL，用于稀释样品和其它试剂；
(2)XOD溶液：取25U/2.6mL的XOD，用75mM的PB溶液稀释至0.08U/mL的XOD工作液，用移液枪吹匀，置于冰上保存，待用；
(3)底物配制：精密称取适量黄嘌呤(XA)加入0.1N NaOH溶液5mL中，超声溶解，后者中加入75mM PB溶液95mL，配制终浓度为0.48mM的底物母液，涡旋混匀1min，每次实验前新鲜配制；
(4)供试药物配制：精密称取受试药物适量，用DMSO溶解配制为10mM的储备液，于-20℃避光保存。实验前用PB稀释至不同浓度(0～100mM)，DMSO含量小于0.1％。
11.2实验步骤
(1)在96孔板上加入不同浓度的待测样品溶液100μL，再加入0.08U/mL XOD 50μL，并以相同体积的PB作为空白对照，以别嘌呤醇(Allopurinol)为阳性对照，在酶标仪上37℃孵育3min，每组平行设置4个复孔。
(2)加入底物0.48mM XA 50μL启动反应，在波长295nm处每隔15s读数一次，记录吸光值，共计7min。数据处理：使用Excel分析处理数据，并用GraphPad Prism6.0.2计算得到半数抑制浓度(IC50)。
(3)测定结果如表1所示，合成得到的多数苯并呋喃酰胺化衍生物均具有较显著地黄嘌呤氧化酶抑制活性。
表1苯并呋喃类酰胺化衍生物对黄嘌呤氧化酶的抑制活性
Figure PCTCN2017071614-appb-000053
a IC50值均为四次平行实验的均结果；
b文献报道：IC50＝2.55μM；
实施例12
采用DPPH自由基清除实验对合成得到的苯并呋喃类酰胺化衍生物进行抗氧化活性评价。
12.1试剂与标准溶液的配制
(1)DPPH溶液的配制：精密称取适量DPPH，加入MeOH超声溶解，配制10mM储备液，-20℃保存。实验前用MeOH稀释至0.1mM，避光保存；
(2)供试药物的配制：精密称取受试药物适量，用MeOH溶解配制10m储备液，-20℃避光保存。实验前用MeOH稀释至不同浓度(0～100mM)。
12.2实验步骤
(1)在96孔板上加入不同浓度的待测样品溶液100μL，再加入0.1mM DPPH溶液100μL，并以相同体积的MeOH作为空白对照，以槲皮素(Quercetin)作为阳性对照，在酶标仪上37℃摇晃混匀1min后，暗处放置30min，每组平行设置3个复孔。
(2)在波长517nm处用酶标仪记录其吸光值，数据处理：使用Excel分析处理数据，并用GraphPad Prism 6.0.2计算得到半数抑制浓度(IC50)。
(3)测定结果如表2所示，大部分芳基苯并呋喃衍生物均具有较明显的抗氧化作用，由其结构可推测邻酚羟基对其清除自由基能力有着重要影响。
表2苯并呋喃类酰胺化衍生物抗氧化活性
Figure PCTCN2017071614-appb-000054
a IC50值均为三次平行实验的均结果；
b文献报道IC50＝9.1μM；
实施例13
采用细胞模型评价合成得到的苯并呋喃类酰胺化衍生物的抗氧化活性评价(清除超氧阴离子的能力)。
哺乳动物巨噬细胞在吞噬细菌、衰老变性细胞、免疫复合物或氧化低密度脂蛋白后，吞噬细胞进入功能活化状态，细胞内溶酶体功能增强、ROS水平增加，分泌合成炎症性细胞因子。脂多糖(Lipopolysaccharide,LPS)是革兰阴性菌细胞壁的主要成分，也是其致病的主要物质基础，可通过刺激单核/巨噬细胞产生并释放大量的活性氧(主要包括O2￣·和H2O2)、一氧化氮(NO)、IL-1β以及肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)等炎症因子参与机体急性期应答，引起机体炎性损伤。本发明利用LPS刺激巨噬细胞模型来评价苯并呋喃类酰胺化衍生物在细胞水平清除超氧阴离子的抗氧化活性。
13.1试剂与标准溶液的配制
(1)待测药物：精密称取受试药物适量，用DMSO配制为10mM的样品储备液，并于-20℃保存备用。
(2)LPS：精密称取造模剂LPS适量，用DMEM配制为0.1μg/mL的样品储备液，并于-20℃保存备用。
(3)探针稳定剂DTPA：精密称取DTPA适量，用DMSO配制为20mM的样品储备液，并用PB稀释成100μM，于4℃保存备用。
(4)HE：在厌氧、避光条件下用二甲基亚砜溶解探针配制成20mmol/L的储备溶液，于-80℃条件下避光保存。使用前取出，用DMEM配制成10μmol/L溶液，置于冰上避光保存备用。
13.2实验步骤
LPS刺激巨噬细胞RAW 264.7造模并给药：
(1)RAW 264.7细胞接种于培养皿(60mm)，每皿4mL，置于饱和湿度，37℃的5％CO2孵箱中培养。实验分为空白组、模型组和给药组，其中模型组和给药组分别加入造模剂LPS(以DMEM配制为)，实验优选的LPS浓度为0.1μg/mL，造模时间24h，以维生素E为阳性对照。
(2)细胞培养12h后，吸弃去上清液，加入含0.1μg/mL LPS培养基配置的终浓度为10μM的待测药物，每个待测样品分别设两个平行复孔。
(3)继续置于细胞培养箱中培养24h后，弃去上清液，用PBS冲洗细胞，加入含有探针HE(10μM)的DMEM培养基，避光培养30min。
(4)随后用冰冷的PBS/(100μM)DTPA冲洗细胞两次，再加入1mL PBS/(100μM)DTPA，用刮刀将细胞分为两份(700μL和300μL)分别收集至EP管中，4℃，13000rpm离心10min，去除上清液，将细胞团块置于-80℃保存待用。其中前者含700μL细胞液用于检测物2-OH-E+的LC-MS含量测定，后者含300μL细胞液用于细胞蛋白浓度的测定。
13.3LC-MS分析条件、待测样品前处理及数据处理方法
(1)LC-MS色谱条件：仪器：Agilent 1100series LC-QMS；色谱柱：Agilent Zorbax SB-Aq(4.6×50mm，1.8μm)；流动相：0.1％甲酸-水(A相)；0.1％甲酸-乙腈(B)；梯度条件：0～2min，5％～30％B；2～5min，30％B；5～7min，30％～100％B；7～10min，100％B；流速：1mL/min；MS条件：positive ion mode，SIM(SIM1：1.2～3min[M+H]+288(加兰他敏)；3～10min[M]+330(2-OH-E+))；裂解电压：120V；氮气流速：10.0L/min；
(2)待测样品前处理：每个EP管中加入500μL乙腈，避光，超声(10s，八个循环，100W)，并于4℃，12,000g离心10min后，取其上清液转移到1.5mL EP管中，真空干燥1h,得红色干燥物并用100μL质谱水复溶样品，同时加入内标物加兰他敏，使其待测样品终浓度为0.1μM。待测样品于4℃，13,000rpm离心10min，进LC-MS分析。
(3)超氧阴离子清除率的计算：通过峰面积/蛋白浓度来评价产物2-OH-E+的生成量。化合物对XOD抑制率或O2-·的清除率按以下公式计算：O2-·自由基清除率＝[Ablank-Asample]/A blank×100％，其中Ablank为对照组2-OH-E+的峰面积值/蛋白浓度；Asample为样品组2-OH-E+的峰面积值/蛋白浓度。
13.4数据结果分析：结果如表3所示，与模型组相比，2-芳基苯并呋喃类酰胺化衍生物5a，5b，5c，6b，6c，6d，6e及Salvianolic acid C对LPS刺激巨噬细胞RAW 264.7产生的超氧阴离子有较明显的清除作用。与Ve相比，衍生物5a，5b，5c，6b，6d和6e表现出更好的超氧阴离子清除活性。显示出芳基苯并呋喃类酰胺化衍生物具有较好的抗氧化活性，尤其是衍生物5a，5b，5c，6b，6d和6e表现出优于Ve的超氧阴离子清除活性。
表3芳基苯并呋喃类酰胺化衍生物对RAW 264.7中超氧阴离子的清除活性评价
Figure PCTCN2017071614-appb-000055
实施例14
采用细胞模型评价苯并呋喃类酰胺化衍生物对尿酸生成的影响。
(1)供试药物配制：精密称取受试药物适量，用DMSO配制为10mM的样品储备液，并于4℃保存备用。
(2)实验步骤：正常肝细胞LO2接种于6孔板中培养(铺板密度1×106个/mL)，铺板4h后，进行细胞实验。实验分为空白组、模型组和给药组，其中模型组和给药组分别加入造模剂黄嘌呤(以DMSO配制为储备液待用)，空白组加入相同体积DMSO，并以非布索坦(Febuxostat)作为阳性对照。阳性对照药非布索坦取5μM，受试药物用培养基稀释至所需浓度(10μM，30μM，50μM)后，加入6孔板中，37℃度孵育15min后，模型组、给药组分别加入终浓度为10μM的黄嘌呤，每组分别设两个复孔，置于细胞培养箱中培养24h后，收集培养基100μL进行LC-MS分析。
(3)质谱分析前处理：取100μL收集的培养基，加入MeOH 400μL、CHCl3 100μL和H2O 300μL，涡旋混匀，20000g离心10min，取上层浓缩干燥。H2O 100μL复溶，并加入内标加兰他敏(1.25μM)10μL(终浓度为0.125μM)，13000rpm离心10min，取上层进行LC-MS分析。
(3)测定结果如表4所示，芳基苯并呋喃类酰胺化衍生物5b，6e和Salvianolic acid C(1)对由黄嘌呤造模形成的肝细胞模型中尿酸的生成有较为明显的抑制作用，并呈现出一定的浓度依赖性。
表4苯并呋喃类酰胺化衍生物5b，6e对细胞模型中尿酸的抑制作用
Figure PCTCN2017071614-appb-000056
实施例15
采用氧嗪酸钾诱导的高尿酸血症动物模型评价苯并呋喃类酰胺化衍生物对小鼠血尿酸的影响
(1)实验动物：昆明小鼠由扬州大学实验动物中心提供。给药前12h及试验期间禁食，实验过程中不限制饮水。
(2)供试药物配制：精密称取受试药物适量，用相应体积的生理盐水超声溶解至溶液澄清，配制为储备液，并于4℃保存备用。
(3)实验步骤：小鼠随机分为9组，每组10只：①正常对照组，②模型对照组，③衍生物5b低剂量组：20mg/kg，④衍生物5b中剂量组：40mg/kg，⑤衍生物5b高剂量组：80mg/kg，⑥衍生物6e低剂量组：20mg/kg，⑦衍生物6e中剂量组：40mg/kg，⑧衍生物6e高剂量组：80mg/kg，⑨阳性对照组：别嘌醇20mg/kg。供试药物用0.5％羧甲基纤维素钠(CMC-Na)配制成适宜浓度，正常组小鼠腹腔注射0.5％羧甲基纤维素钠，其余各组小鼠腹腔注射氧嗪酸钾300mg/kg。1h后正常组和模型组小鼠腹腔注射0.5％羧甲基纤维素钠，其余各组腹腔注射供试药物10mL/kg。给药2h后摘眼球取血500μL，室温放置1h，3000rpm离心5min，取上层血清于-4℃保存。按尿酸检测试剂盒说明书方法测定小鼠血清中尿酸水平。同时取小鼠肝脏，生理盐水漂洗，取100mg，加900μL预冷的生理盐水匀浆，离心，取上清按试剂盒说明书方法测定肝脏匀浆中黄嘌呤氧化酶活力。
(4)测定结果如表5所示：小鼠腹腔注射氧嗪酸钾后，血中尿酸水平显著升高，与正常组相比，有显著性差异，表明造模成功。给药后，血清中的尿酸水平和肝脏中XOD活性均与浓度呈现一定的剂量依赖性。
表5衍生物5b，6e对氧嗪酸钾所致小鼠高尿酸血症模型的影响
Figure PCTCN2017071614-appb-000057
a给药组与模型组相比：*P<0.05，**P<0.01。
实施例16
为初步阐明所合成的系列2-芳基苯并呋喃类酰胺化衍生物与黄嘌呤氧化酶可能的结合模式及相互作用位点，采用分子对接的方法(Molecular docking experiment)初步对2-芳基苯并呋喃类酰胺化衍生物6e,6b，5b和Salvianolic acid(1)与黄嘌呤氧化酶进行了作用机制研究。黄嘌呤氧化酶(PDB code:1FIQ)由A、B、C三条亚基构成，其中包含有三个活性结构域，分别为A链中的Fe/S中心结构域，B链中的FAD结构域和C链中的钼蝶呤结构域。在对接实验中，使用Febuxostat和Allopurinol这两个已知黄嘌呤氧化酶抑制剂作为对照。分析分子对接结果，根据打分函数确定，认为化合物单体与黄嘌呤氧化酶结合能最低的对接构象为其优势构象。对接结果如图3：化合物1,6e,6b,5b及Febuxostat和Allopurinol的对接打分分别为-7.1,-7.42,-7.25,-7.1,-8.19,-5.21。由图3可以看出，化合物1,6e,6b和Febuxostat与多个相同的氨基酸残基发生相互作用，如Lys249，Lys256，Leu257，Val259，Ile264，Ile353，Arg394和Leu404，表明化合物1,6e,6b可能与阳性药Febuxostat相似，结合于黄嘌呤氧化酶相同的结构域而发挥着抑制该酶的活性。由图3还可以看出，衍生物5b则与氨基酸残基Pro1076，Phe1009，Thr1010，Arg880，Val1011，Leu1014，Glu802，Ser876，Glu879发挥着相互作用，且与残基Phe1009,Thr1010and Glu879形成三个氢键，这与阳性药Allopurinol相似，可能作用于黄嘌呤氧化酶共同的结构域而发挥着酶抑制作用。同时还观察出，氢键的形成和疏水性相互作用对于维系蛋白受体-配体构象的稳定发挥重要作用。
在图3活性化合物与黄嘌呤氧化酶分子对接实验结果中，(A)Febuxostat(B)Allopurinol(C)6b(D)6e(E)Salvianolic acid C(1)(F)5b。
实施例17
衍生物5b的制剂
(1)本发明的衍生物5b的片剂：
衍生物5b2mg，淀粉88g，硬脂酸镁3g
制备工艺：取本发明的衍生物5b过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗粒，干燥，压片，即得。
(2)本发明的衍生物5b的胶囊
衍生物5b2mg，淀粉88g，硬脂酸镁3g
制备工艺：取本发明的衍生物5b过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗
粒，干燥，装胶囊，即得。
(3)本发明的衍生物5b的软胶囊
衍生物5b 10mg，大豆卵磷脂100g
制备工艺：取本发明的衍生物5b，加大豆软磷脂，胶体磨混匀，抽真空，压制，即得软胶囊。
(4)本发明的衍生物5b的冻干粉：
衍生物5b2g，亚硫酸钠4g，乙醇50mL，加水定容至1000mL；
制备工艺：取本发明的衍生物5b分散在乙醇中，亚硫酸钠溶于水中，在超声或搅拌条件下将拿硫酸钠溶液逐渐加入，使成澄清透明溶液；补加水定容至足量；经0.22μm微孔滤膜过滤，冷冻干燥得到。
实施例18
衍生物6e的制剂
(1)本发明的衍生物6e的片剂：
衍生物6e2mg，淀粉88g，硬脂酸镁3g
制备工艺：取本发明的衍生物6e过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗粒，干燥，压片，即得。
(2)本发明的衍生物6e的胶囊
衍生物6e2mg，淀粉88g，硬脂酸镁3g
制备工艺：取本发明的衍生物6e过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗
粒，干燥，装胶囊，即得。
(3)本发明的衍生物6e的软胶囊
衍生物6e10mg，大豆卵磷脂100g
制备工艺：取本发明的衍生物6e，加大豆软磷脂，胶体磨混匀，抽真空，压制，即得软胶囊。
(4)本发明的衍生物6e的冻干粉：
衍生物6e2g，亚硫酸钠4g，乙醇50mL，加水定容至1000mL；
制备工艺：取本发明的衍生物6e分散在乙醇中，亚硫酸钠溶于水中，在超声或搅拌条件下将拿硫酸钠溶液逐渐加入，使成澄清透明溶液；补加水定容至足量；经0.22μm微孔滤膜过滤，冷冻干燥得到。
上述实施例为本发明具有代表性的实施方案的举例说明，仅仅是示例性的说明，但本发明的实施方式并不受上述实施例的限制，其他任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。

Claims (8)

  1. 一种芳基苯并呋喃类酰胺化衍生物，其结构通式如式I所示：
    Figure PCTCN2017071614-appb-100001
    R1，R2，R3，R4，R5，R6，R7，R8各自独立为氢，羟基，卤素，硝基，苄基，未取代或经卤素、羟基、硝基、C1-2烷氧基中选出1～3个取代基取代的C1-4烷基，未取代或经卤素、羟基、硝基、C1-2烷氧基中选出1～2个取代基取代的C1-3烷氧基；
    R9为氢，未取代或经卤素、羟基、巯基、C1-3烷基、C1-2烷氧基中选出1～3个取代基取代的C1-4烷基，未取代或经卤素、羟基取代的3-乙基-1H-吲哚，未取代或经卤素、羟基、C1-2烷氧基取代的苯基，未取代或经卤素、羟基、C1-2烷氧基取代的苄基，未取代或经卤素、羟基、C1-2烷氧基取代的其他芳基(其中，所述卤素X＝F，Cl，Br，n＝1，2，3)，如3,4-二羟基苄基，3,4-二甲氧基苄基，4-羟基苄基，4-甲氧基苄基等；
    R10为氢、—COOH、—COOCH3、—COOCH2CH3、—COOCH(CH3)2、—COO(CH2)2CH3或经卤素、羟基、C1-3烷基、C1-2烷氧基中选出1～3个取代基取代的C1-4烷基。
  2. 根据权利要求1的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：
    Figure PCTCN2017071614-appb-100002
    其中：R3，或、和、R4为羟基以外的其他取代基；
    其中：R9，或、和、R10为氢以外的其他取代基；
    或；一种芳基苯并呋喃类酰胺化衍生物：
    结构通式如下任意一项所示：
    Figure PCTCN2017071614-appb-100003
    Figure PCTCN2017071614-appb-100004
    结构通式如式XI所示：
    Figure PCTCN2017071614-appb-100005
    结构通式如式XII所示：
    Figure PCTCN2017071614-appb-100006
    或；一种芳基苯并呋喃类酰胺化衍生物：
    结构通式如式1所示：
    Figure PCTCN2017071614-appb-100007
    其中R1，R2，R3各自独立为未取代或经卤素、羟基取代的C1-4烷基，未取代或经卤素、羟基取代的C1-3烷氧基，卤素，硝基，苄基；
    R4为氢，未取代或经卤素、羟基、巯基取代的C1-4烷基，3-乙基-1H-吲哚，未取代或经卤素、羟基取代的苯基，未取代或经卤素、羟基取代的苄基，未取代或经卤素、羟基取代的其他芳基(其中，卤素X＝F，Cl，Br)，如3,4-二羟基苄基，3,4-二甲氧基苄基，4-羟基苄基， 4-甲氧基苄基等；
    R5为氢，—COOH、—COOCH3、—COOCH2CH3、—COOCH(CH3)2、—COO(CH2)2CH3或者为氢、C1-4烷基；
    或；一种芳基苯并呋喃类酰胺化衍生物：
    Figure PCTCN2017071614-appb-100008
    其中R1，R2，R3各自独立为氢，羟基；
    R4为氢；R5为氢；
    或；一种芳基苯并呋喃类酰胺化衍生物：
    Figure PCTCN2017071614-appb-100009
    其中R1，R2，R3各自独立为未取代或经卤素、羟基取代的C1-4烷基，未取代或经卤素、羟基取代的C1-3烷氧基，卤素，硝基，苄基；
    R4为氢，未取代或经卤素、羟基、巯基取代的C1-4烷基，3-乙基-1H-吲哚，未取代或经卤素、羟基取代的苯基，未取代或经卤素、羟基取代的苄基，未取代或经卤素、羟基取代的其他芳基(其中，卤素X＝F，Cl，Br)，如3,4-二羟基苄基，3,4-二甲氧基苄基，4-羟基苄基，4-甲氧基苄基等；
    R5为氢，—COOH、—COOCH3、—COOCH2CH3、—COOCH(CH3)2、—COO(CH2)2CH3或者为氢、C1-4烷基；
    或；一种芳基苯并呋喃类酰胺化衍生物：
    Figure PCTCN2017071614-appb-100010
    其中R1，R2，R3各自独立为未取代或经卤素、羟基取代的C1-4烷基，未取代或经卤素、羟基取代的C1-3烷氧基，卤素，硝基，苄基；
    R4为未取代或经卤素、羟基、巯基取代的C1-4烷基，3-乙基-1H-吲哚，未取代或经卤素、羟基取代的苯基，未取代或经卤素、羟基取代的苄基，未取代或经卤素、羟基取代的其他芳基(其中，卤素X＝F，Cl，Br)，如3,4-二羟基苄基，3,4-二甲氧基苄基，4-羟基苄基，4-甲氧基苄基；
    R5为—COOH、—COOCH3、—COOCH2CH3、—COOCH(CH3)2、—COO(CH2)2CH3或者为氢、C1-4烷基；
    或；一种芳基苯并呋喃类酰胺化衍生物：
    结构通式如下任意一项所示：
    Figure PCTCN2017071614-appb-100011
    Figure PCTCN2017071614-appb-100012
    Figure PCTCN2017071614-appb-100013
    Figure PCTCN2017071614-appb-100014
    Figure PCTCN2017071614-appb-100015
  3. 根据权利要求1-2的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：其用于制备抗氧化、制备清除自由基、制备黄嘌呤氧化酶抑制剂、治疗痛风、或、治疗高尿酸血症的组合物、药物、保健品上的应用。
  4. 根据权利要求1-2的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：合成方法如下：
    (1)将丹酚酸C与无机碱混合，超声溶解于混合溶剂中，加热不断检测反应，待反应完全后，硅胶柱色谱分离，得到中间体Tournefolic acid A；
    (2)将SOCl2冰浴冷却下加入有机溶剂中，反应30min后，加入丙氨酸、苯丙氨酸、半胱氨酸、酪氨酸、甲硫氨酸、色氨酸、D/L-DOPA等氨基类化合物，室温搅拌，浓缩蒸干溶剂并洗涤干燥，得系列羧基保护的氨基衍生物；
    (3)将中间产物Tournefolic acid A在缩合剂作用下分别与系列氨基衍生物以摩尔比1：1.1～2混合，溶于有机溶剂中，室温搅拌，反应后处理后经硅胶柱色谱分离，得到系列的芳基苯并呋喃类酰胺化衍生物；
    (4)在系列芳基苯并呋喃类酰胺化衍生物中加入无机碱，混合溶剂超声溶解，室温搅拌或加热回流，TLC不断监测反应，样品混合物经后处理，再经过硅胶柱色谱或制备型高效液相色谱分离纯化，得到系列高纯度的苯并呋喃类酰胺化衍生物。
  5. 根据权利要求4的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：合成方法如下：
    所述的第1步中的无机碱水解可选择LiOH，NaOH，KOH等，有机溶媒为THF，MeOH，H2O或为其混合溶剂，优选为混合溶剂MeOH/H2O，且溶剂体积比优选为3：1～5：1，反应时间优选为8～12h；
    所述的第1步中的分离纯化方法具体为：硅胶柱层析，用氯仿/甲醇/甲酸(10：1：0.1，v/v/v)等度洗脱得到中间体Tournefolic acid A；
    所述的第2步中的氨基类化合物为各种D/L构型的氨基酸及其它手性氨基衍生物，有机溶剂为甲醇、乙醇及异丙醇等试剂，反应时间优选为18～24h；
    所述的第2步中的分离纯化方法具体为：甲醇和乙醚分别多次交互洗涤样品并浓缩蒸干；
    所述的第3步中的缩合剂可以为DCC/HoBt，EDCI/HoBt，HATU，HBTU，PyBOP，优 选为EDCI/HoBt，有机溶剂优选为混合溶剂DMF/CH2Cl2(3：1～5：1v/v)，反应时间优选为8～12h；
    所述的第3步中的分离纯化方法具体为：硅胶柱层析，氯仿/甲醇梯度洗脱得目标衍生物；
    所述的第4步中的无机碱可以为LiOH，NaOH，KOH，优选为NaOH，有机溶媒为THF，MeOH，H2O或为其混合溶剂，优选为MeOH/H2O(3：1～5：1v/v)，反应时间优选为8～12h。
  6. 根据权利要求4的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：分离纯化方法如下：
    硅胶柱层析分离纯化，流动相为氯仿/甲醇/甲酸梯度洗脱；或制备型液相色谱柱分离纯化样品混合物，其中，色谱柱为(Agilent，Zorbax-C18，5μm，9.4×250mm)，色谱条件优选为：流速：8mL/min，检测波长：281nm，柱温：30℃，流动相：乙腈-0.1％甲酸-水。
  7. 根据权利要求4的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：
    L-多巴甲酯盐酸盐的结构是：
    Figure PCTCN2017071614-appb-100016
  8. 根据权利要求4的一种芳基苯并呋喃类酰胺化衍生物，其特征在于：
    L-多巴甲酯盐酸盐的制备方法为：准确量取无水甲醇5mL于25mL圆底烧瓶中，冰水浴冷却15min，缓慢逐滴加入SOCl2(493μL，6.780mmol)，0℃搅拌30min后，加入准确称取的L-DOPA(800mg，4.520mmol)，放置至室温，搅拌反应24h，待反应完全后，蒸干溶剂MeOH和反应剩余的SOCl2，用MeOH和Et2O多次交互洗涤样品，浓缩蒸干，得反应产物L-多巴甲酯盐酸盐。
PCT/CN2017/071614 2016-02-03 2017-01-19 一种芳基苯并呋喃类酰胺化衍生物及医药用途 Ceased WO2017133464A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US16/075,370 US11021454B2 (en) 2016-02-03 2017-01-19 Type of aryl benzofuran amidated derivative and medical use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610076098.8 2016-02-03
CN201610076098.8A CN105859667A (zh) 2016-02-03 2016-02-03 一种芳基苯并呋喃类酰胺化衍生物及医药用途

Publications (1)

Publication Number Publication Date
WO2017133464A1 true WO2017133464A1 (zh) 2017-08-10

Family

ID=56624196

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/071614 Ceased WO2017133464A1 (zh) 2016-02-03 2017-01-19 一种芳基苯并呋喃类酰胺化衍生物及医药用途

Country Status (3)

Country Link
US (1) US11021454B2 (zh)
CN (1) CN105859667A (zh)
WO (1) WO2017133464A1 (zh)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112305114A (zh) * 2020-10-29 2021-02-02 诺峰药业(成都)有限公司 一种测定二氟泼尼酯眼用乳剂中山梨酸含量的高效液相色谱方法
CN116183799A (zh) * 2022-12-30 2023-05-30 山东京博生物科技有限公司 一种三氯氧磷中氯化亚砜的含量分析方法及其应用

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105859667A (zh) 2016-02-03 2016-08-17 中国药科大学 一种芳基苯并呋喃类酰胺化衍生物及医药用途
CN106928173B (zh) * 2017-03-07 2019-04-16 中国科学院新疆理化技术研究所 薰衣草中的酚酸类化合物及其制备方法和用途
CN108997282B (zh) * 2018-07-25 2022-04-29 安徽工程大学 具有α-葡萄糖苷酶抑制活性的芳基苯并呋喃类衍生物
CN115403544B (zh) * 2022-09-16 2024-05-31 台州永健医药科技有限公司 一种丹酚酸c的制备方法
CN116283855B (zh) * 2023-02-10 2025-09-23 上海仲时药业有限公司 丹酚酸c衍生物及其制备方法和医药用途
CN117069630B (zh) * 2023-08-01 2025-08-29 上海仲时药业有限公司 一种ros响应前药和逐级靶向纳米药物递送系统及其制备方法和应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102423310A (zh) * 2012-01-04 2012-04-25 中国药科大学 丹酚酸c在制备防治高尿酸血症药物中的应用
CN105859667A (zh) * 2016-02-03 2016-08-17 中国药科大学 一种芳基苯并呋喃类酰胺化衍生物及医药用途

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1778292A (zh) 2004-11-26 2006-05-31 中国科学院上海药物研究所 一类丹参酚酸的医学用途

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102423310A (zh) * 2012-01-04 2012-04-25 中国药科大学 丹酚酸c在制备防治高尿酸血症药物中的应用
CN105859667A (zh) * 2016-02-03 2016-08-17 中国药科大学 一种芳基苯并呋喃类酰胺化衍生物及医药用途

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
TANG, HONGJIN ET AL.: "Synthesis and evaluation of xanthine oxidase inhibitory and antioxidant activities of 2-arylbenzo[b]furan derivatives based on salvianolic acid C", EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY, vol. 124, 31 August 2016 (2016-08-31), pages 637 - 648, XP055403557, ISSN: 0223-5234 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112305114A (zh) * 2020-10-29 2021-02-02 诺峰药业(成都)有限公司 一种测定二氟泼尼酯眼用乳剂中山梨酸含量的高效液相色谱方法
CN116183799A (zh) * 2022-12-30 2023-05-30 山东京博生物科技有限公司 一种三氯氧磷中氯化亚砜的含量分析方法及其应用

Also Published As

Publication number Publication date
US20190040029A1 (en) 2019-02-07
CN105859667A (zh) 2016-08-17
US11021454B2 (en) 2021-06-01

Similar Documents

Publication Publication Date Title
WO2017133464A1 (zh) 一种芳基苯并呋喃类酰胺化衍生物及医药用途
JPH10245337A (ja) 薬学的組成物
CN116947756B (zh) 环丁烯二酮基喹啉类化合物及其制备方法、药物组合物和应用
WO2021223718A1 (zh) 醛基类化合物及其制备方法、药物组合物和用途
AU2016253911A1 (en) Carboxylic acid URAT1 inhibitor containing diarylmethane structure, preparation method and use thereof
WO2024082629A1 (zh) 哌嗪取代苯酚类衍生物及其用途
KR101682427B1 (ko) 통증 및 기타 질병을 치료하기 위한 화합물 및 방법
CN111635309B (zh) 一种新型解热镇痛药物及其制备方法和用途
CN112679353B (zh) 一种基于丁苯酞结构的前体药物或其药学上可接受的金属盐及其制备方法和应用
CN111635315A (zh) 一种解热镇痛药物及其制备方法和应用
WO2009143715A1 (zh) 类维甲酸衍生物及其药物组合物与用途
CN113461563B (zh) Nqo1激活型6-重氮基-5-氧代-l-正亮氨酸前药及其制备方法和应用
CN1295229C (zh) 紫杉烷衍生物的结晶及其制备方法
CN114364659A (zh) 嘧啶-5-甲酰胺化合物
JP4789618B2 (ja) 新規転写因子、その製法および用途
CN107586284A (zh) 一种2‑芳基苯并呋喃类衍生物在制备痛风药物中的用途
CN115124531A (zh) 一类4-氮杂色胺酮芳香硫醚衍生物、制备方法及应用
CN102653514B (zh) 绿原酸类似物及其制备方法和应用
WO2007051408A1 (fr) Composes destines a la prevention ou au traitement des infections a h. pylori, procedes de preparation et utilisation
CN114181193B (zh) 一种阿魏酸衍生物及其制备方法和应用
CN120058667B (zh) 一种布洛芬-α-硫辛酸共晶及其制备方法与应用
JP7260912B2 (ja) リファガールの類縁体、及びリファガール又はその類縁体を含む多標的キナーゼ阻害剤
TW202448441A (zh) 脯氨醯羥化酶抑制劑及其用途
WO2026007900A1 (zh) 一类万古霉素衍生物、其制备方法、药物组合物和用途
WO2026092351A1 (zh) 一种苯基丙烯酰胺类化合物及其制备方法和应用

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: 17746784

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17746784

Country of ref document: EP

Kind code of ref document: A1