CN116655577B - Flavone derivative aminopeptidase N inhibitor and preparation method and application thereof - Google Patents

Flavone derivative aminopeptidase N inhibitor and preparation method and application thereof Download PDF

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CN116655577B
CN116655577B CN202310930238.3A CN202310930238A CN116655577B CN 116655577 B CN116655577 B CN 116655577B CN 202310930238 A CN202310930238 A CN 202310930238A CN 116655577 B CN116655577 B CN 116655577B
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曹江营
邓志鹏
刘玉红
马艳
董文亮
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Shandong University of Traditional Chinese Medicine
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    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
    • C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
    • C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
    • C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
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    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
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Abstract

The invention belongs to the technical field of synthesis and medical application of organic compounds, and particularly relates to a flavone derivative aminopeptidase N inhibitor, a preparation method and application thereof. The flavone derivative aminopeptidase N inhibitor has a structure shown in a formula (I):formula (I). In-vitro aminopeptidase N inhibition activity tests show that the enzyme inhibition activity of partial flavone derivative aminopeptidase N inhibitors is improved by one to two orders of magnitude compared with that of positive medicine ubenimex; the enzyme inhibition activity of individual flavone derivative aminopeptidase N inhibitor reaches nanomolar level, solves the problems of poor activity and single structure of the non-peptide aminopeptidase N inhibitor, and enriches the structural diversity of the aminopeptidase N inhibitor.

Description

Flavone derivative aminopeptidase N inhibitor and preparation method and application thereof
Technical Field
The invention belongs to the technical field of synthesis and medical application of organic compounds, and particularly relates to a flavone derivative aminopeptidase N inhibitor, a preparation method and application thereof.
Background
Aminopeptidase N (APN, CD13, EC 3.4.11.2) is a zinc ion-dependent type II transmembrane protein,belonging to the family of metalloproteinases, anchored to the surface of the cell membrane in the form of homodimers, in the form of "hippocampus". The catalytic domain is positioned outside the cell, can hydrolyze basic or neutral amino acid at the N end of the substrate, and participates in degradation of the polypeptide substrate. The catalytic active center of aminopeptidase N contains 1 zinc ion and S 1 、S 1 ’、S 2 ' three hydrophobic pockets. The zinc ions are combined with the zinc ion chelating groups in the substrate, and the three hydrophobic pockets are respectively combined with P of the substrate 1 、P 1 ' and P 2 A' site. Studies have shown that aminopeptidase N plays an important role in tumor formation, invasive metastasis and tumor angiogenesis.
Aminopeptidase N has the following effects: (1) aminopeptidase N is involved in tumor metastasis. The main components of the extracellular matrix with barrier function are collagen, fibronectin, laminin and the like, and the components are crosslinked into a net structure to support a tissue structure, so that the extracellular matrix plays an important role in stability and signal transmission among cells. Aminopeptidase N on the surface of the tumor cells can hydrolyze type IV collagen of the collagen family, so that the tumor cells break through the basement membrane barrier and enter blood vessels or lymphatic vessels to transfer. (2) aminopeptidase N is involved in tumor neovascularization. Blood vessels in tumor tissue are more abundant and irregular than normal tissue. The normal endothelial cell surface does not express aminopeptidase N, and the endothelial cell surface in the angiogenesis stage has high expression of aminopeptidase N, and participates in the endothelial cell morphogenesis process. Aminopeptidase N inhibitors do not affect endothelial cell proliferation, but can inhibit tubular structure formation during morphogenesis. (3) Aminopeptidase N is one of the surface markers of liver cancer stem cells (J Clin Invest 2010,120 (9), 3326-3339). The presence of tumor stem cells is an important cause of tumor resistance, recurrence and metastasis. The research shows that the aminopeptidase N inhibitor and the fluorouracil and other cytotoxic antitumor drugs are combined together to have synergistic antitumor effect. The aminopeptidase N inhibitor is expected to solve the problems of drug resistance, recurrence and metastasis of liver cancer by inhibiting liver cancer stem cells.
Aminopeptidase N inhibitors are classified according to structure into peptoid inhibitors and nonpeptidid inhibitors. Ubenimex is a drug marketed in the eighties of the 20 th century, is a peptoid aminopeptidase N inhibitor, has a dipeptide-like structure, and is used as an immunopotentiator for the adjuvant treatment of leukemia. In addition, reported natural peptoid aminopeptidase N inhibitors include amastatin (CAS No. 67655-94-1), probestin (CAS No. 123652-87-9), phebestin (CAS No. 187402-73-9), and the like. There are various aminopeptidases, carboxypeptidases and dipeptidyl peptidases in the body, and peptoid inhibitors have poor selectivity. The peptoid inhibitors have a plurality of chiral centers, the chiral difficulty is high in control during the synthesis process, and the total synthesis cost is high. Therefore, the development of aminopeptidase N inhibitors which are not peptide and are easy to synthesize has great significance. At present, the non-peptide aminopeptidase N inhibitor also has the problems of poor activity, single structure and the like. Flavonoid compounds widely exist in traditional Chinese medicines, are one of the material bases for the traditional Chinese medicines to exert the drug effect, and are all flavonoid compounds such as chrysin, quercetin and the like. The flavonoid compound has relatively wide pharmacological action, low toxicity and low synthesis difficulty, and is suitable for serving as a structural fragment of a medicament. Therefore, the development of the flavonoid aminopeptidase N inhibitor has better scientific research value.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a flavone derivative aminopeptidase N inhibitor, a preparation method and application thereof.
To achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, a flavone derivative aminopeptidase N inhibitor has a structure represented by formula (I):
formula (I);
wherein:
R 1 is that、/>、/>、/>、/>、/>、、/>、/>、/>、/>、/>、/>、、/>、/>、/>、/>、/>、/>Or (b)；
R 2 Is H, methoxy,、/>、/>Or (b)。
In a second aspect, a method for preparing a flavone derivative aminopeptidase N inhibitor according to the first aspect, wherein R 2 Is H, methoxy,、/>Or->In the process, the intermediate shown in the formula (II) reacts with methyl bromoacetate in the presence of sodium hydride to generate an intermediate shown in the formula (III), the intermediate shown in the formula (III) is hydrolyzed in a potassium hydroxylamine methanol solution, and then acidized to generate the flavone derivative aminopeptidase N inhibitor;
formula (II); />Formula (III);
wherein: r is R 3 Is H, hydroxy, methoxy,Or->；R 4 Is H,Methoxy, & gt>Or->。
Preferably, the intermediate shown in the formula (II) is obtained by reacting the intermediate shown in the formula (VII) with dimethyl sulfate under the condition of potash property;
formula (VII).
Preferably, when R 3 Is thatIn the process, an intermediate shown in a formula (II) is obtained by reacting an intermediate shown in a formula (VII) with 4- (2-bromoethyl) morpholine hydrobromide under the condition of potash property;
Formula (VII).
Preferably, when R 3 Is thatThe intermediate shown in the formula (II) reacts with methyl bromoacetate under the alkaline condition of potassium carbonate, and then is hydrolyzed under the alkaline condition and condensed with morpholine to obtain the intermediate shown in the formula (VII);
formula (VII).
Preferably, when R 3 In the case of H, 2, 6-dihydroxylAcetophenone and carboxylic acid R 1 Generating an intermediate shown in a formula (X) by COOH reaction, and generating an intermediate shown in a formula (XI) by generating Baker-Venkataraman rearrangement of the intermediate shown in the formula (X) under alkaline conditions; in a glacial acetic acid/concentrated sulfuric acid system, performing cyclization reaction on an intermediate shown in a formula (XI) to generate an intermediate shown in a formula (II);
formula (X); />Formula (XI).
In a third aspect, a method for preparing the flavone derivative aminopeptidase N inhibitor according to the first aspect, wherein, when R 2 Is thatReacting an intermediate shown in a formula (IV) with methyl bromoacetate in the presence of sodium hydride to generate an intermediate shown in a formula (V), hydrolyzing the intermediate shown in the formula (V) in a potassium hydroxylamine methanol solution, then acidifying to generate an intermediate shown in a formula (VI), and removing tert-butoxycarbonyl groups from the intermediate shown in the formula (VI) under the acidic condition of trifluoroacetic acid to generate the flavone derivative aminopeptidase N inhibitor; an intermediate shown in a formula (IV) is obtained by reacting an intermediate shown in a formula (VII) with N-tert-butoxycarbonyl-2-bromoethylamine under a potash condition;
Formula (IV); />Formula (V);formula (VI); />Formula (VII).
Preferably, when R 3 In the case of hydroxyl, the intermediate represented by the formula (II) or the intermediate represented by the formula (VII) is prepared by the following preparation method:
2,4, 6-Trihydroxyacetophenone and carboxylic acid R 1 Generating an intermediate shown in a formula (VIII) by COOH reaction, and generating an intermediate shown in a formula (IX) by generating Baker-Venkataraman rearrangement of the intermediate shown in the formula (VIII) under alkaline conditions; in a glacial acetic acid/concentrated sulfuric acid system, performing cyclization reaction on an intermediate shown in a formula (IX) to generate an intermediate shown in a formula (II) or a formula (VII);
formula (VIII); />Formula (IX).
In a fourth aspect, the use of a flavone derivative aminopeptidase N inhibitor according to the first aspect, and a pharmaceutically acceptable salt, solvate or prodrug thereof, in the manufacture of a medicament for the prophylaxis or treatment of a condition associated with aberrant aminopeptidase activity, including hematological or solid tumours, inflammatory conditions, multiple sclerosis, tissue ulceration or ulceration conditions, periodontal disease, epidermolysis bullosa or malaria.
In a fifth aspect, a pharmaceutical composition suitable for oral or parenteral administration, comprising a flavone derivative aminopeptidase N inhibitor according to the first aspect, or a pharmaceutically acceptable salt, solvate or prodrug thereof, and one or more pharmaceutically acceptable carriers or excipients.
The terms and definitions used in the present invention have the following meanings:
by "pharmaceutically acceptable salt" is meant the therapeutically effective and non-toxic salt form of the compound of formula (I). Many such salts are known in the art. A cationic salt formed on any acidic group (e.g., carboxyl) or an anionic salt formed on any basic group (e.g., amino). Many of these salts are known in the art, such as cationic salts including salts of alkali metals (e.g., sodium and potassium) and alkaline earth metals (e.g., magnesium and calcium) and organic salts (e.g., amine salts). Anionic salts can also be obtained by using the corresponding acids, including inorganic acids such as sulfuric acid, nitric acid, phosphoric acid, hydrochloric acid, and the like; or organic acids such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, 2-hydroxypropionic acid, 2-oxopropionic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, cyclohexylsulfinic acid, 2-hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid, and the like. These salts are well known to the skilled artisan and the skilled artisan can prepare any salt provided by the knowledge in the art. In addition, the skilled artisan can take one salt and another salt based on solubility, stability, ease of formulation, etc. Determination and optimization of these salts is within the experience of the skilled artisan.
A "solvate" is a complex formed by the combination of a solute (e.g., an aminopeptidase N inhibitor) and a solvent (e.g., water). See "J. Honig et al, the Van Nostrand Chemist's Dictionary, p.650 (1953)". Pharmaceutically acceptable solvents for use in the present invention include those solvents that do not interfere with the biological activity of the aminopeptidase N inhibitor (e.g., water, ethanol, acetic acid,N,NDimethylformamide, dimethyl sulfoxide, solvents known or readily ascertainable to those skilled in the art).
The compounds of formula (I) may also exist in other protected forms or derivatives, which are obvious to a person skilled in the art and are intended to be included within the scope of the present invention.
The substituents described above may themselves also be substituted with one or more substituents. Such substituents include those listed in "C.Hansch and A.Leo, substituent Constants for Correlation Analysis in Chemistry and Biology (1979)". Preferred substituents include, for example, alkyl, alkenyl, alkoxy, hydroxy, oxo, nitro, amino, aminoalkyl (e.g., aminomethyl, etc.), cyano, halo, carboxy, carbonylalkoxy (e.g., carbonylaethoxy, etc.), thio, aryl, cycloalkyl, heteroaryl, heterocycloalkyl (e.g., piperidinyl, morpholinyl, pyrrolyl, etc.), imino, hydroxyalkyl, aryloxy, arylalkyl, and combinations thereof.
The invention has the technical characteristics and beneficial effects that:
the inhibitor of the invention is a flavone derivative aminopeptidase N inhibitor, and belongs to a non-peptide aminopeptidase N inhibitor. In the inhibitor, a hydroxamic acid group chelates zinc ions of an aminopeptidase N catalytic active center, and a flavone mother nucleus structure is combined with a hydrophobic pocket of the aminopeptidase N catalytic active center. The flavonoid compound widely exists in the traditional Chinese medicine, is one of the substance bases for the traditional Chinese medicine to exert the efficacy, has small toxicity and small synthesis difficulty, and is suitable for serving as a structural unit of the medicine. In vitro aminopeptidase N inhibition activity tests show that the enzyme inhibition activity of part of the compounds is improved by one to two orders of magnitude compared with that of positive drug ubenimex; the enzyme inhibition activity of the individual compounds reaches the nanomolar level, the problems of poor activity and single structure of the non-peptide aminopeptidase N inhibitor are solved, and the structural diversity of the aminopeptidase N inhibitor is enriched; the invention takes flavonoid compounds widely existing in traditional Chinese medicines as a structural parent nucleus, and has better drug property.
The inhibitor provided by the invention has the advantages of simple preparation route, strong feasibility, higher industrial value, less pollution of three wastes and more environmental friendliness. The synthesis of the inhibitor mainly relates to the following steps: 1) Synthesizing a mother nucleus with a flavone structure; 2) Introduction of side chains. Esterifying 2, 6-dihydroxyacetophenone or 2,4, 6-trihydroxyacetophenone with corresponding carboxylic acid; then, generating a Baker-Venkataraman rearrangement of the generated intermediate under alkaline conditions to generate a 1, 3-diketone intermediate; the intermediate is cyclized under glacial acetic acid/sulfuric acid system to produce mother nucleus with flavone structure. The introduction of the side chain involves nucleophilic substitution reaction, nucleophilic addition elimination reaction, acidification reaction, etc. These reactions are all conventional reactions, simple to operate and highly feasible.
The total synthesis method of the flavone mother nucleus generally includes a chalcone method and a beta-propanedione method (Baker-Venkataraman method). The chalcone method is to synthesize chalcone first and then close ring in the presence of oxidant or dehydrogenation catalyst to synthesize flavone. Hydroxy chalcone is sensitive to oxidizing agents and is not suitable for preparing hydroxy flavone. The beta-propanedione method adopted by the invention is subjected to three steps of esterification, rearrangement and ring closure, and the synthesis method is mild, the operation is simple, and the yield is higher.
The hydroxamic acid can be prepared by condensing carboxyl and hydroxylamine hydrochloride by a condensing agent such as EDCI, DCC, TBTU, but the ester bond must be hydrolyzed to obtain carboxylic acid, and the product obtained by condensation must be purified by column chromatography; the ester bond is hydrolyzed by potassium hydroxylamine to obtain the hydroxamic acid group, the steps are shortened, the separation and purification are simple, and the column chromatography purification is not needed.
Drawings
FIG. 1 is a schematic diagram showing a process for producing the objective compound (6) according to the present invention;
FIG. 2 is a schematic diagram showing a process for preparing the target compound (12), the target compound (15) and the target compound (19) according to the present invention;
FIG. 3 is a schematic diagram showing a process for producing the objective compound (22) and the objective compound (27) according to the present invention.
Detailed Description
The invention will be further illustrated with reference to specific examples, but is not limited thereto.
Meanwhile, the experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, are commercially available.
The preparation of the potassium hydroxylamine methanol solution in each example of the invention is as follows: 28.00g of potassium hydroxide and 23.35g of hydroxylamine hydrochloride are respectively dissolved in 70mL and 120mL of redistilled methanol to obtain a solution A and a solution B; dropwise adding the solution A into the solution B, and stirring for 40 minutes in an ice water bath; filtering to obtain new potassium hydroxylamine methanol solution.
Example 1 preparation of Compound (6)
R in Compound (6) 1 And the corresponding compound numbers are shown in table 1.
Table 1 Compound No. and R 1
N-hydroxy-2- ((2- (3-iodophenyl) -4-oxo-4)H-benzopyran-5-yl) oxy) acetamide (6 l) as shown in figure 1, by the following procedure:
1) Preparation of 2-acetyl-1, 3-phenylenebis (3-iodobenzoate) (2 l)
3-iodobenzoic acid (39.16 g,157.90 mmol) and 2, 6-dihydroxyacetophenone (1) (12.00 g,78.94 mmol) were dissolved in pyridine (200 mL). Phosphorus oxychloride (30.20 g,197.00 mmol) was added dropwise at 0deg.C, and after 20 minutes the ice bath was removed and reacted at 25deg.C for 12 hours. After the completion of the reaction, the reaction solution was poured into 300mL of water and allowed to stand for 12 hours. Suction filtering, washing the filter cake with water for 2-3 times. Drying at 45-50 ℃ to obtain 45.17 g intermediate (2 l). Yield: 94%. ESI-MS: m/z [ M+H ] ] + =613.1. The chemical formula of intermediate (2 l) is shown below:
。
2) Preparation of 1- (2, 6-dihydroxyphenyl) -3- (3-iodophenyl) propane-1, 3-dione (3 l)
Potassium tert-butoxide (11.10 g,99.11 mmol) was dissolved in 70mLN,NDimethylformamide, nitrogen protection, dropwise addition of intermediate (2 l) (20.21 g,33.02 mmol)N,N80mL of dimethylformamide solution was reacted at 25℃for 1.5 hours. The pH was adjusted to neutral with phosphoric acid, the reaction solution was poured into 300mL of water, and extracted 3 times with 80mL each time of ethyl acetate. The organic phases were combined and washed 3 times with 80mL of saturated aqueous sodium bicarbonate solution and 3 times with 70mL of saturated aqueous sodium chloride solution. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Column chromatography purification, petroleum ether-ethyl acetate as mobile phase, in a volume ratio of 5:1, afforded 8.20g of intermediate (3 l). Yield: 65%. The chemical formula of intermediate (3 l) is shown below:
。
3) 5-hydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (4 l)
Intermediate (3L) (2.00 g,5.23 mmol) was added to 15mL of glacial acetic acid, heated to 90 ℃, 150. Mu.L of concentrated sulfuric acid was added, the temperature was raised to 120℃and the reaction was carried out for 1 hour. The reaction solution was cooled to 25℃and poured into 300mL of water, and allowed to stand for 10 hours, whereby a solid was precipitated. Filtering, washing the residue with 100mL saturated sodium bicarbonate aqueous solution to neutrality, and oven drying at 45-50deg.C. Column chromatography purification, with methylene chloride-methanol as mobile phase at a volume ratio of 100:1, afforded 1.03g of intermediate (4 l). Yield: 54%. ESI-MS: m/z [ M+H ] ] + =365.2. The chemical formula of intermediate (4 l) is shown below:
。
4) Methyl 2- ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy) acetate (5 l)
Intermediate (4 l) (1.00 g,2.75 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.13 g,3.25 mmol), and the mixture was reacted at 25℃for 3 minutes, followed by methyl bromoacetate (0.63 g,4.12 mmol) and the reaction was carried out at 25℃for 8 hours. The reaction solution was poured into 200mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purification by column chromatography, with methylene chloride-methanol as the mobile phase at a volume ratio of 60:1, afforded 1.01g of intermediate (5 l). Yield: 84%. ESI-MS: m/z [ M+H ]] + = 437.2. The chemical formula of intermediate (5 l) is shown below:
。
5）N-hydroxy-2- ((2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-5-yl) oxy) acetamide (6 l)
0.50g of the intermediate (5 l) was added to 5mL of potassium hydroxylamine methanol solution, reacted at 25℃for 0.5 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 6, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days, to give 0.46g of the objective compound (6 l). Yield: 91, mp 216-218 ℃. The nuclear magnetic data of the obtained product are as follows ： 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.01 (s, 1H), 9.18 (s, 1H), 8.42 (t,J= 1.7 Hz, 1H), 8.11 (dt,J= 7.8 Hz,J= 1.2 Hz, 1H), 7.97 (dt,J= 7.8 Hz,J= 1.2 Hz, 1H), 7.76 (t,J= 8.4 Hz, 1H), 7.44 (d,J= 8.4 Hz, 1H), 7.38 (t,J= 7.8 Hz, 1H), 7.05-7.03 (m, 2H), 4.72 (s, 2H); 13 C NMR (151 MHz, DMSO-d 6 ) δ 175.93, 162.46, 158.01, 155.63, 155.02, 138.74, 133.24, 132.71, 131.20, 129.46, 124.10, 112.18, 109.81, 108.05, 107.36, 93.95, 66.47。
Preparation of the compounds (6 a) - (6 k) and (6 m) - (6 u), e.g.preparation of (6 l), except for R used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of (6 l).
Prepared byN-hydroxy-2- ((4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetamide (6 a) has the following nuclear magnetic data: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.06 (s, 1H), 9.18 (s, 1H), 8.11-8.09 (m, 2H), 7.77 (t,J= 8.4 Hz, 1H), 7.65-7.57 (m, 3H), 7.41 (d,J= 8.4 Hz, 1H), 7.04 (d,J= 8.2 Hz, 1H), 7.00 (s, 1H), 4.73 (s, 2H)。
prepared 2- ((2- (3-bromophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NThe nuclear magnetic data of hydroxyacetamide (6 c) are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (d,J= 1.7 Hz, 1H), 9.19 (d,J= 1.5 Hz, 1H), 8.30 (t,J= 1.9 Hz, 1H), 8.13-8.10 (m, 1H), 7.83-7.75 (m, 2H), 7.55 (t,J= 8.0 Hz, 1H), 7.45 (d,J= 8.4 Hz, 1H), 7.09 (s, 1H), 7.04 (d,J= 8.0 Hz, 1H ), 4.73 (s, 2H)。
prepared 2- ((2- (3-chlorophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NThe nuclear magnetic data of hydroxyacetamide (6 i) are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.00 (s, 1H), 9.21 (s, 1H), 8.07 (s, 1H), 7.99 (d,J= 7.7 Hz, 1H), 7.72 (t,J= 8.3 Hz, 1H), 7.62 (d,J= 8.0 Hz, 1H), 7.56 (t,J= 8.0 Hz, 1H), 7.36 (d,J= 8.3 Hz, 1H), 6.99 (d,J= 8.1 Hz, 2H), 4.70 (s, 2H); 13 C NMR (101 MHz, DMSO-d 6 ) δ 177.42, 164.01, 159.46, 157.08, 156.50, 134.74, 134.00, 132.65, 131.43, 130.84, 125.78, 124.83, 113.66, 111.27, 109.49, 108.97, 67.98。
prepared 2- ((2- (3-fluorophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NThe nuclear magnetic data of hydroxyacetamide (6 k) are as follows: 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.20 (s, 1H), 7.97-7.94 (m, 2H), 7.76 (t,J= 8.3 Hz, 1H), 7.65-7.61 (m, 1H), 7.48-7.45 (m, 1H), 7.42 (d,J= 8.4 Hz, 1H), 7.07 (s, 1H), 7.04 (d,J= 8.2 Hz, 1H), 4.73 (s, 2H)。
prepared byN-hydroxy-2- (4-oxo-2- (m-tolyl-4)H-benzopyran-5-yl) oxy) acetamide (6 m) has the following nuclear magnetic data: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 9.21 (s, 1H), 7.84-7.81 (m, 2H), 7.72 (t,J= 8.3 Hz, 1H), 7.45-7.33 (m, 3H), 7.00 (d,J= 8.1 Hz, 1H), 6.89 (s, 1H), 4.70 (s, 2H), 2.39 (s, 3H); 13 C NMR (101 MHz, DMSO-d 6 ) δ 177.49, 164.06, 161.31, 157.16, 156.54, 138.49, 134.61, 132.44, 130.45, 128.91, 126.53, 123.35, 113.69, 111.22, 109.48, 108.10, 68.06, 20.90。
prepared byN-hydroxy-2- ((2- (naphthalen-1-yl) -4-oxo-4)H-benzopyran-5-yl) oxy) acetamide (6 s) has the following nuclear magnetic data: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.06 (s, 1H), 9.19 (s, 1H), 8.19 (d,J= 8.2 Hz, 1H), 8.12-8.07 (m, 2H), 7.89 (d,J= 7.1 Hz, 1H), 7.78 (t,J= 8.3 Hz, 1H), 7.70-7.63 (m, 3H), 7.30 (d,J= 8.4 Hz, 1H), 7.10 (d,J= 8.3 Hz, 1H), 6.65 (s, 1H), 4.77 (s, 2H)。
prepared 2- ((2- (furan-2-yl) -4-oxo-4)HBenzopyran-5-yl) oxy-NThe nuclear magnetic data of hydroxyacetamide (6 t) are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.00 (s, 1H), 9.17 (s, 1H), 8.07 (d,J= 1.6 Hz, 1H), 7.75 (t,J= 8.4 Hz, 1H), 7.44 (d,J= 3.5 Hz, 1H), 7.30 (d,J= 8.4 Hz, 1H), 7.04 (d,J= 8.2 Hz, 1H), 6.83 (dd,J= 3.6 Hz,J= 1.7 Hz, 1H), 6.61 (s, 1H), 4.72 (s, 2H)。
Prepared byN-hydroxy-2- ((4-oxo-2- (thiophen-2-yl) -4)H-benzopyran-5-yl) oxy) -acetamide (6 u) has the following nuclear magnetic data: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 9.18 (s, 1H), 8.05-7.99 (m, 2H), 7.74 (t,J= 8.3 Hz, 1H), 7.32-7.30 (m, 2H), 7.04 (d,J= 8.2 Hz, 1H), 6.86 (s, 1H), 4.72 (s, 2H)。
EXAMPLE 2 preparation of Compound (12)
The compound (12) comprises a compound (12 a) and a compound (12 b), and the compound (12 a) has the chemical formula of
；
The chemical formula of the compound (12 b) is
。
2,2' - ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5, 7-diyl) bis (oxy))NThe preparation of hydroxyacetamide) (12 b), as shown in fig. 2, proceeds as follows:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b)
3-iodobenzoic acid (59.76g,240.97 mmol) and 2,4, 6-trihydroxyacetophenone (7) (14.88 g,80.00 mmol) were dissolved in 200mL pyridine. Phosphorus oxychloride (42.84 g,279.09 mmol) was added dropwise at 0deg.C, and after 20 minutes the ice bath was removed and reacted at 25deg.C for 12 hours. After the completion of the reaction, the reaction solution was poured into 300mL of water and allowed to stand for 12 hours. Suction filtering, washing the filter cake with water for 2-3 times. Drying at 45-50 ℃ to obtain an intermediate (8 b). ESI-MS: m/z [ M+H ]] + = 859.2. Chemistry of intermediate (8 b)The formula is as follows:
。
2) Preparation of 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b)
Potassium tert-butoxide (17.92 g,160.00 mmol) was dissolved in 70mLN,NDimethylformamide, nitrogen protection, dropwise addition of intermediate (8 b) (34.28 g,39.95 mmol)N,N80mL of dimethylformamide solution was reacted at 25℃for 1.5 hours. The pH was adjusted to neutral with phosphoric acid, the reaction solution was poured into 300mL of water, and extracted 3 times with 80mL each time of ethyl acetate. The organic phases were combined and washed 3 times with 80mL of saturated aqueous sodium bicarbonate solution and 3 times with 70mL of saturated aqueous sodium chloride solution. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 5:1, so as to obtain an intermediate (9 b). The chemical formula of intermediate (9 b) is shown below:
。
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b)
Intermediate (9 b) (3.97 g,9.97 mmol) was added to 15mL of glacial acetic acid, heated to 90 ℃, 150. Mu.L of concentrated sulfuric acid was added, the temperature was raised to 120 ℃, and the reaction was carried out for 1 hour. The reaction solution was cooled to 25℃and poured into 300mL of water, and allowed to stand for 10 hours, whereby a solid was precipitated. Filtering, washing the residue with 100mL saturated sodium bicarbonate aqueous solution to neutrality, and oven drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 100:1, so as to obtain an intermediate (10 b). ESI-MS: m/z [ M+H ] ] + = 381.2. The chemical formula of intermediate (10 b) is shown below:
。
4) Dimethyl 2,2' - ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-like compoundsPreparation of 5, 7-diyl) bis (oxy)) diacetate (11 b)
Intermediate (10 b) (2.00 g,5.26 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.46 g,11.5 mmol), and the mixture was reacted at 25℃for 3 minutes, followed by methyl bromoacetate (2.41 g,15.75 mmol) and the mixture was reacted at 25℃for 8 hours. The reaction solution was poured into 200mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (11 b). ESI-MS: m/z [ M+H ]] + =525.5. The chemical formula of intermediate (11 b) is shown below:
。
5) 2,2' - ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5, 7-diyl) bis (oxy))NPreparation of-hydroxyacetamide) (12 b)
Intermediate (11 b) (0.70 g,1.33 mmol) was added to 5mL of potassium hydroxylamine in methanol, reacted at 25℃for 0.5 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 6, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days, to give 0.60g of the objective compound (12 b). Yield: 86, mp 218-220 ℃. The nuclear magnetic data of the obtained product are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.05 (s, 1H), 10.96 (s, 1H), 9.20 (s, 1H), 9.12 (s, 1H), 8.38 (d,J= 2.0 Hz, 1H), 8.07 (d,J= 7.9 Hz, 1H), 7.95 (d,J= 7.8 Hz, 1H), 7.36 (t,J= 7.8 Hz, 1H), 7.01-6.94 (m, 2H), 6.67 (d,J= 2.3 Hz, 1H), 4.70 (s, 1H), 4.67 (s, 1H); 13 C NMR (101 MHz, DMSO-d 6 ) δ 177.10, 164.42, 163.94, 163.04, 159.58, 159.07, 158.19, 140.66, 134.55, 133.20, 131.51, 125.99, 109.24, 109.11, 99.67, 96.05, 95.95, 68.51, 66.67。
Preparation of Compound (12 a), e.g. preparation of Compound (12 b), except R used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (12 b).
Prepared 2,2' - ((4-oxo-2-phenyl-4)HBenzopyran-5, 7-diyl) bis (oxy))N-hydroxyacetamide) (12 a) the nuclear magnetic data are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.08 (s, 1H), 10.94 (s, 1H), 9.17 (s, 1H), 9.08 (s, 1H), 8.08-8.06 (m, 2H), 7.64-7.56 (m, 3H), 6.97 (d,J= 2.3 Hz, 1H), 6.93 (s, 1H), 6.68 (d,J= 2.3 Hz, 1H), 4.70 (s, 2H), 4.66 (s, 2H)。
EXAMPLE 3 preparation of Compound (15)
The compound (15) comprises a compound (15 a), a compound (15 b) and a compound (15 c), and the compound (15 a) has the chemical formula of；/>
The chemical formula of the compound (15 b) is；
The chemical formula of the compound (15 c) is。
N-hydroxy-2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)H-benzopyran-5-yl) oxy) acetamide (15 b), as shown in fig. 2, by the following procedure:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b) was performed as in example 2.
2) 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b) was prepared as in example 2.
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b) was carried out as in example 2.
4) 5-hydroxy-2- (3-iodophenyl) -7-methoxy-4HPreparation of benzopyran-4-one (13 b)
Intermediate (10 b) (0.80 g,2.11 mmol) was dissolved in 10mLN,NDimethyl formamide, dimethyl sulfate (0.40 g,3.17 mmol) and potassium carbonate are added0.38g,2.75 mmol). The reaction was carried out at 80℃for 1h. The reaction solution was poured into 100mL of water. Standing for 10h. Filtering. Drying at 45-50deg.C. 10mL ethyl acetate was slurried for 20 minutes. Filtering. Intermediate (13 b) is obtained. The chemical formula of intermediate (13 b) is shown below:
。
5) Methyl 2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)H-benzopyran-5-yl) oxy) acetate (14 b) preparation
Intermediate (13 b) (0.40 g,1.02 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.05 g,1.25 mmol), and the mixture was reacted at 25℃for 3 minutes, followed by methyl bromoacetate (0.23 g,1.50 mmol) and the mixture was reacted at 25℃for 8 hours. The reaction solution was poured into 50mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (14 b). The chemical formula of intermediate (14 b) is shown below:
。
6）N-hydroxy-2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)HPreparation of benzopyran-5-yl) oxy) acetamide (15 b)
Intermediate (14 b) (0.10 g,0.21 mmol) was added to 2mL of potassium hydroxylamine in methanol, reacted at 25℃for 0.5 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 6, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days to give 0.082g of the objective compound (15 b). Yield: 84, mp 216-218 ℃. The nuclear magnetic data of the obtained product are as follows: 1 H NMR (600 MHz, DMSO-d 6 ) δ 11.06 (s, 1H), 9.13 (s, 1H), 8.41 (t,J= 1.8 Hz, 1H), 8.09 (dt,J= 8.0 Hz,J= 1.2 Hz, 1H), 7.96 (dt,J= 7.8 Hz,J= 1.3 Hz, 1H), 7.36 (t,J= 7.8 Hz, 1H), 7.04 (d,J= 2.3 Hz, 1H), 6.95 (s, 1H), 6.61 (d,J= 2.3 Hz, 1H), 4.69 (s, 2H), 3.92 (s, 3H); 13 C NMR (151 MHz, DMSO-d 6 ) δ 176.06, 163.59, 163.39, 158.48, 158.25, 157.15, 139.57, 133.49, 132.20, 130.41, 124.95, 108.12, 107.68, 98.21, 94.95, 94.17, 67.46, 55.67。
Preparation of Compounds (15 a), (15 c) as described for the preparation of Compound (15 b), except that R is used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (15 b).
Prepared byN-hydroxy-2- ((7-methoxy-4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetamide (15 a) has the following nuclear magnetic data: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.10 (s, 1H), 9.15 (s, 1H), 8.10-8.07 (m, 2H), 7.61-7.56 (m, 3H), 6.99 (d,J= 2.2 Hz, 1H), 6.91 (s, 1H), 6.62 (d,J= 2.2 Hz, 1H), 4.70 (s, 2H), 3.92 (s, 3H)。
EXAMPLE 4 preparation of Compound (19)
The compound (19) comprises a compound (19 a) and a compound (19 b), and the compound (19 a) has the chemical formula of
，
The chemical formula of the compound (19 b) is
。
2- ((7- (2-aminoethoxy) -2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NThe preparation of hydroxyacetamide trifluoroacetate salt (19 b), as shown in fig. 2, comprises the following steps:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b) was performed as in example 2.
2) 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b) was prepared as in example 2.
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b)Example 2 was prepared.
4) Tert-butyl (2- ((5-hydroxy-2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-7-yl) oxy) ethyl) carbamate (16 b)
Intermediate (10 b) (1.00 g,2.63 mmol) was dissolved in 20mLN,N-dimethylformamide, added toNBoc-2-bromoethylamine (0.71 g,3.17 mmol), potassium carbonate (0.47 g,3.41 mmol), potassium iodide (0.57 g,3.43 mmol) and tetrabutylammonium bromide (0.42 g,1.30 mmol). The reaction was carried out at 80℃for 2 hours. The reaction solution was poured into 100mL of water. Standing for 10 hours. Filtering. Drying at 45-50deg.C. Column chromatography, mobile phase: dichloromethane-tetrahydrofuran-methanol, the volume ratio of the three is 100:40:0.7, and the intermediate (16 b) is obtained. ESI-MS: m/z [ M+H ]] + =524.3. The chemical formula of intermediate (16 b) is shown below:
。
5) Methyl 2- ((7- (2-tert-Butoxycarbonyl) amino) ethoxy) -2- (3-iodophenyl) -4-oxo-4H-benzopyran-5-yl) oxy) acetate (17 b) preparation
Intermediate (16 b) (1.20 g,2.29 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.11 g,2.75 mmol), and the mixture was reacted at 25℃for 3 minutes, followed by methyl bromoacetate (0.53 g,3.46 mmol) and the reaction was carried out at 25℃for 8 hours. The reaction solution was poured into 50mL of water and allowed to stand for 10 hours. Ethyl acetate was extracted 3 times, 50mL each. The organic phases are combined. 100mL of saturated aqueous sodium chloride solution was washed 1 time. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 70:1, so as to obtain an intermediate (17 b). ESI-MS: m/z [ M+H ] ] + = 596.3. The chemical formula of intermediate (17 b) is shown below:
。
6) Tert-butyl (2- ((5- (2- (hydroxylamine) -2-oxyethoxy) -2- (3-iodobenzene)Phenyl) -4-oxo-4HPreparation of benzopyran-7-yl) oxy) ethyl) carbamate (18 b)
Intermediate (17 b) (0.38 g,0.64 mmol) was added to 5mL of potassium hydroxylamine in methanol, reacted at 25℃for 1 hour, and the reaction solution was poured into 50mL of water. Adjusting pH to 6 with 5% citric acid aqueous solution to obtain white solid, standing for 10 hr, filtering, and naturally drying for 2 days to obtain intermediate (18 b). ESI-MS: m/z [ M+H ]] + = 597.5. The chemical formula of intermediate (18 b) is shown below:
。
7) 2- ((7- (2-aminoethoxy) -2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NPreparation of hydroxyacetamide trifluoroacetate salt (19 b)
Intermediate (18 b) (0.15 g,0.25 mmol) was added to 3mL of dichloromethane followed by 3mL of trifluoroacetic acid. The reaction was carried out at 25℃for 3 hours. The solvent was concentrated off, 10mL ethyl acetate was added and filtered. After drying naturally for 2 days, 0.12g of the objective compound (19 b) was obtained. Yield: 78%. The nuclear magnetic data of the obtained product are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.06 (s, 1H), 9.21 (s, 1H), 8.41 (t,J= 1.8 Hz, 1H), 8.17 (s, 3H), 8.10 (dt,J= 8.1 Hz,J= 1.2 Hz, 1H), 7.97 (dt,J= 7.8 Hz,J= 1.2 Hz, 1H), 7.37 (t,J= 7.9 Hz, 1H), 7.10 (d,J= 2.3 Hz, 1H), 6.98 (s, 1H), 6.59 (d,J= 2.3 Hz, 1H), 4.70 (s, 2H), 4.35 (t,J= 4.9 Hz, 2H), 3.31 (s, 2H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 177.10, 164.38, 163.02, 159.64, 159.25, 158.46, 158.29, 140.72, 134.59, 133.22, 131.52, 126.04, 116.08, 109.29, 109.17, 99.47, 96.08, 96.00, 68.45, 65.87, 38.60。
preparation of Compound (19 a) as described for the preparation of Compound (19 b), except that R is used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (19 b).
Prepared 2- ((7- (2-amino)Ethoxy) -4-oxo-2-phenyl-4HBenzopyran-5-yl) oxy-N-nuclear magnetic data of hydroxyacetamide trifluoroacetate (19 a) as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.31 (s, 3H), 8.08-8.06 (m, 2H), 7.63-7.56 (m, 3H), 7.02 (d,J= 2.3 Hz, 1H), 6.91 (s, 1H), 6.59 (d,J= 2.3 Hz, 1H), 4.70 (s, 2H), 4.36 (t,J= 4.9 Hz, 2H), 3.32 (t,J= 5.1 Hz, 2H). 13 C NMR (101 MHz, DMSO-d 6 ) δ 177.22, 164.43, 162.96, 161.26, 159.21, 159.03, 158.29, 132.24, 131.03, 129.56, 126.57, 119.14, 109.08, 108.52, 99.29, 95.93, 68.46, 65.80, 38.57。
example 5
The compound (22) comprises a compound (22 a), and the compound (22 a) has the chemical formula of
。
N-hydroxy-2 ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetamide (22 a) as shown in fig. 3, by the following procedure:
1) 5-hydroxy-7- (2-morpholinoethoxy) -2-phenyl-4HPreparation of benzopyran-4-one (20 a)
By varying R used 1 COOH was benzoic acid, and intermediate (10 a) was obtained by reference to the preparation method in example 2. Intermediate (10 a) (1.50 g,5.91 mmol) was dissolved in 20mLN,NDimethylformamide, 4- (2-bromoethyl) morpholine hydrobromide (1.95 g,7.09 mmol), potassium carbonate (1.96 g,14.2 mmol), potassium iodide (0.49 g,2.95 mmol) and tetrabutylammonium bromide (0.48 g,1.49 mmol) were added. The reaction was carried out at 80℃for 2 hours. The reaction solution was poured into 100mL of water and allowed to stand for 6 hours. Filtering. Drying at 45-50deg.C to obtain intermediate (20 a). ESI-MS: m/z [ M+H ] ] + =368.4. The chemical formula of intermediate (20 a) is shown below:
。
2) Methyl 2- ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetate (21 a)
Intermediate (20 a) (1.20 g,3.27 mmol) was dissolved in 15mLN,NTo dimethylformamide, 60% sodium hydride (0.16 g,4.00 mmol) was added in portions and reacted at 25℃for 3 minutes. Methyl bromoacetate (0.75 g,4.90 mmol) was added. The reaction was carried out at 25℃for 8 hours. Pour into 100mL of water. Ethyl acetate was extracted 3 times, 50mL each. The organic phases are combined. And (5) drying the anhydrous magnesium sulfate. Filtering. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 50:1, thus obtaining the intermediate (21 a). ESI-MS: m/z [ M+H ]] + =440.5. The chemical formula of intermediate (21 a) is shown below:
。
3）N-hydroxy-2 ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetamide (22 a)
Intermediate (21 a) (0.50 g,1.14 mmol) was added to 8mL of potassium hydroxylamine in methanol. The reaction was carried out at 25℃for 1 hour. The reaction was poured into 50mL of water. And adjusting the pH of the phosphoric acid aqueous solution with the volume fraction of 2% to 6-7. Standing for 10 hours. Filtering. Natural drying gave 0.36g of compound (22 a). Yield: 72%. The nuclear magnetic data of the obtained product are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 9.22 (s, 1H), 8.06-8.04 (m, 2H), 7.62-7.55 (m, 3H), 6.96 (d,J= 2.3 Hz, 1H), 6.87 (s, 1H), 6.59 (d,J= 2.2 Hz, 1H), 4.69 (s, 2H), 4.23 (t,J= 5.7 Hz, 2H), 3.61 (t,J= 4.6 Hz, 4H), 2.73 (t,J= 5.7 Hz, 2H), 2.50 (s, 4H); 13 C NMR (101 MHz, DMSO-d 6 ) δ 177.23, 164.53, 163.73, 161.13, 159.23, 158.23, 132.16, 131.09, 129.53, 126.53, 108.67, 108.43, 99.35, 95.65, 68.56, 66.63, 57.16, 54.01。
Example 6
The compound (27) includes a compound (27 a), and the compound (27 a) has the chemical formula of
。
N-hydroxy-2- ((7- (2-morpholin-2-oxyethyl) -4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetamide (27 a) as shown in fig. 3, by the following procedure:
preparation of methyl 2- ((5-hydroxy-4-oxo-2-phenyl-4H-benzopyran-7-yl) oxo) acetate (23 a)
By varying R used 1 COOH was benzoic acid, and intermediate (10 a) was obtained by reference to the preparation method in example 2. Intermediate (10 a) (4.00 g,15.75 mmol) was dissolved in 50mLN,NTo dimethylformamide was added potassium carbonate (2.82 g,20.43 mmol) and methyl bromoacetate (2.89 g, 18.89). The reaction was carried out at 50℃for 1 hour. The reaction solution was poured into 200mL of water. Standing for 5 hours. Filtering. Drying at 45-50 ℃ to obtain an intermediate (23 a). ESI-MS: m/z [ M+H ]] + =327.4. The chemical formula of intermediate (23 a) is shown below:
。
2) 2- ((5-hydroxy-4-oxo-2-phenyl-4)H-benzopyran-7-yl) oxy) acetic acid (24 a) preparation
Intermediate (23 a) (4.90 g,15.03 mmol) was added to 100mL of methanol, and potassium hydroxide (5.28 g,94.29 mmol) was added. The reaction was carried out at 65℃for 1 hour. The reaction solution was poured into 200mL of water, and the pH was adjusted to be neutral with a 10% volume fraction of phosphoric acid aqueous solution. Standing for 1 hour. Filtering. Drying at 45-50 ℃ to obtain an intermediate (24 a). The chemical formula of intermediate (24 a) is shown below:
。
3) 5-hydroxy-7- (2-morpholin-2-yloxyethoxy) -2-phenyl-4HPreparation of benzopyran-4-one (25 a)
Intermediate (24 a) (1.50 g,4.81 mmol) was dissolved in 30mLN,NIn dimethylformamide. 1-Ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.01 g,5.27 mmol), 1-hydroxybenzotriazole (0.71 g,5.26 mmol) and 4-dimethylaminopyridine (0.12 g,0.98 mmol) were added at 0deg.C. Morpholine (0.46 g,5.29 mmol) was added after 30 minutes at 0deg.C. The reaction was carried out at 25℃for 8 hours. The reaction solution was poured into 100mL of water and allowed to stand for 8 hours. Filtering. Drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 200:1, so as to obtain an intermediate (25 a). The chemical formula of intermediate (25 a) is shown below:
。
4) Methyl 2- ((7- (2-morpholin-2-yloxyethoxy) -4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetate (26 a) preparation
Intermediate (25 a) (0.72 g,1.89 mmol) was dissolved in 10mLN,NTo dimethylformamide, 60% sodium hydride (90.72 mg,2.27 mmol) was added in portions. After 3 minutes of reaction at 25℃methyl bromoacetate (0.43 g,2.81 mmol) was added. After reaction at 25℃for 8 hours, the reaction mixture was poured into 100mL of water. Ethyl acetate was extracted 3 times, 50mL each. The organic phases were combined and dried over anhydrous magnesium sulfate. Filtering. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio is 30:1, thus obtaining an intermediate (26 a). ESI-MS: m/z [ M+H ] ] + =454.5. The chemical formula of intermediate (26 a) is shown below:
。
5）N-hydroxy-2- ((7- (2-morpholin-2-oxyethyl) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetamide (27 a)
Intermediate (26 a) (0.50 g,1.10 mmol) was added to 8mL of potassium hydroxylamine in methanol and reacted at 25℃for 1 hour. After the reaction is finished, concentrating the methanol; water was added thereto, and the pH was adjusted to 6-7 with a 10% by volume aqueous phosphoric acid solution, and the mixture was allowed to stand for 12 hours, filtered and naturally dried to obtain 0.32g of Compound (27 a). Yield: 64%. The product obtainedThe nuclear magnetic data of (2) are as follows: 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.12 (s, 1H), 9.18 (s, 1H), 8.08-8.06 (m, 2H), 7.64 -7.56 (m, 3H), 6.98-6.96 (m, 1H), 6.68-6.66 (m, 1H), 6.67 (dd,J= 8.1 Hz,J= 2.3 Hz, 1H), 5.06 (s, 2H), 4.71 (d,J= 3.6 Hz, 2H), 3.68-3.59 (m, 4H), 3.50-3.47 (m, 4H); 13 C NMR (101 MHz, DMSO-d 6 ) δ 177.25, 165.58, 164.51, 163.47, 161.26, 159.17, 158.26, 132.23, 131.13, 129.60, 126.59, 108.93, 108.58, 99.69, 96.00, 68.66, 66.50, 66.43, 45.02, 42.07。
EXAMPLE 7 in vitro Activity assay for inhibiting aminopeptidase N
1) Experimental materials:
porcine renalase N was purchased from Biocol, and the substrate L-leucyl-p-nitroaniline was purchased from Sigma.
Preparation of phosphate buffer: 12.89 g Na (g) 2 HPO 4 . 12H 2 O and 2.18 g NaH 2 PO 4 . 2H 2 O, dissolved in 1000mL deionized water, gave a 50 mM phosphate buffer solution at pH 7.2.
Preparing a substrate solution: l-leucyl-p-nitroaniline is dissolved in dimethyl sulfoxide to prepare 16 mmol/L solution.
Preparing an aminopeptidase N solution: aminopeptidase N was dissolved in the buffer to prepare a solution of 0.15. 0.15 IU/mL.
Preparing an inhibitor solution: the inhibitor is dissolved in dimethyl sulfoxide to prepare stock solution of 100 mmol/L, and then diluted by phosphate buffer solution to prepare solutions with different concentration gradients.
2) The experimental method comprises the following steps:
table 2 composition of the reagents added to each set of experiments
Inhibitor solution, phosphate buffer, substrate solution and enzyme solution were added to 96-well plates in the amounts indicated in the above table. Incubate at 37 ℃ for 0.5 hours. Absorbance was measured at 405, nm and inhibition was calculated according to the following formula:
。
the inhibitor concentration is taken as an abscissa, the inhibition rate is taken as an ordinate, and an origin 9.0 software is used for fitting a curve to obtain the IC 50 Values.
3) Experimental results:
the aminopeptidase N inhibitory activities of the compounds obtained in examples 1 to 6 and ubenimex as a positive drug are shown in Table 3.
TABLE 3 aminopeptidase N inhibitory Activity of different Compounds
[a] The experimental results are expressed as mean ± standard deviation of three independent experiments.
The in vitro enzyme inhibition experiment result shows that: the inhibitory activity of the compounds (6 a), (6 c), (6 i), (6 k), (6 l), (12 a), (12 b), (15 a), (15 b), (19 b) on aminopeptidase N was increased by one to two orders of magnitude compared to the positive control ubenimex. Wherein the compound (12 b) has an optimal aminopeptidase N inhibitory activity.
Example 8
N-hydroxy-2- ((2- (3-iodophenyl) -4-oxo-4)H-benzopyran-5-yl) oxy) acetamide (6 l) as shown in figure 1, by the following procedure:
1) Preparation of 2-acetyl-1, 3-phenylenebis (3-iodobenzoate) (2 l)
3-iodobenzoic acid (39.16 g,157.90 mmol) and 2, 6-dihydroxyacetophenone (1) (12.00 g,78.94 mmol) were dissolved in pyridine (600 mL). Phosphorus oxychloride (24.21 g,157.90 mmol) was added dropwise at 0deg.C, and after 20 minutes the ice bath was removed and reacted for 16 hours at 20deg.C. After the completion of the reaction, the reaction solution was poured into 300mL of water and allowed to stand for 12 hours. Suction filtering, washing the filter cake with water for 2-3 times. Drying at 45-50 ℃ to obtain an intermediate (2 l).
2) Preparation of 1- (2, 6-dihydroxyphenyl) -3- (3-iodophenyl) propane-1, 3-dione (3 l)
Potassium tert-butoxide (7.40 g,66.04 mmol) was dissolved in 70mLN,NDimethylformamide, nitrogen protection, dropwise addition of intermediate (2 l) (20.21 g,33.02 mmol)N,N202.1mL of dimethylformamide solution was reacted at 20℃for 2 hours. The pH was adjusted to neutral with phosphoric acid, the reaction solution was poured into 300mL of water, and extracted 3 times with 80mL each time of ethyl acetate. The organic phases were combined and washed 3 times with 80mL of saturated aqueous sodium bicarbonate solution and 3 times with 70mL of saturated aqueous sodium chloride solution. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 5:1, thus obtaining an intermediate (3 l).
3) 5-hydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (4 l)
Intermediate (3L) (2.00 g,5.23 mmol) was added to 20mL glacial acetic acid, heated to 80 ℃, 100. Mu.L concentrated sulfuric acid was added, the temperature was raised to 110 ℃, and the reaction was carried out for 2 hours. The reaction solution was cooled to 20℃and poured into 300mL of water, and allowed to stand for 10 hours, whereby a solid was precipitated. Filtering, washing the residue with 100mL saturated sodium bicarbonate aqueous solution to neutrality, and oven drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 100:1, thus obtaining an intermediate (4 l).
4) Methyl 2- ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy) acetate (5 l)
Intermediate (4 l) (1.00 g,2.75 mmol) was dissolved in 20mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.11 g,2.75 mmol), and the mixture was reacted at 20℃for 3.5 minutes, followed by addition of methyl bromoacetate (0.42 g,2.75 mmol) and the reaction was carried out at 25℃for 8 hours. The reaction solution was poured into 200mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (5 l).
5）N-hydroxy-2- ((2- (3-iodophenyl) -4-oxo-4) HPreparation of benzopyran-5-yl) oxy) acetamide (6 l)
0.50g of the intermediate (5 l) was added to 5mL of potassium hydroxylamine methanol solution, reacted at 20℃for 1.5 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 6, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days, and the objective compound (6 l) was obtained.
Preparation of the compounds (6 a) - (6 k) and (6 m) - (6 u), e.g.preparation of (6 l), except for R used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of (6 l).
Example 9
N-hydroxy-2- ((2- (3-iodophenyl) -4-oxo-4)H-benzopyran-5-yl) oxy) acetamide (6 l) as shown in figure 1, by the following procedure:
1) Preparation of 2-acetyl-1, 3-phenylenebis (3-iodobenzoate) (2 l)
3-iodobenzoic acid (43.07 g,173.67 mmol) and 2, 6-dihydroxyacetophenone (1) (12.00 g,78.94 mmol) were dissolved in pyridine (120 mL). Phosphorus oxychloride (45.29 g,236.82 mmol) was added dropwise at 5℃for 20 minutes, the ice bath was removed and the reaction was continued for 2 hours at 40 ℃. After the completion of the reaction, the reaction solution was poured into 300mL of water and allowed to stand for 12 hours. Suction filtering, washing the filter cake with water for 2-3 times. Drying at 45-50 ℃ to obtain an intermediate (2 l).
2) Preparation of 1- (2, 6-dihydroxyphenyl) -3- (3-iodophenyl) propane-1, 3-dione (3 l)
Potassium tert-butoxide (11.10 g,99.11 mmol) was dissolved in 70mLN,NDimethylformamide, nitrogen protection, dropwise addition of intermediate (2 l) (20.21 g,33.02 mmol)N,N67mL of dimethylformamide solution and was reacted at 40℃for 0.5 hour. The pH was adjusted to neutral with phosphoric acid, the reaction solution was poured into 300mL of water, and extracted 3 times with 80mL each time of ethyl acetate. The organic phases were combined and washed 3 times with 80mL of saturated aqueous sodium bicarbonate solution and 3 times with 70mL of saturated aqueous sodium chloride solution. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 5:1, thus obtaining an intermediate (3 l).
3) 5-hydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (4 l)
Intermediate (3L) (2.00 g,5.23 mmol) was added to 4mL of glacial acetic acid, heated to 90 ℃, 80. Mu.L of concentrated sulfuric acid was added, the temperature was raised to 120℃and the reaction was carried out for 0.5 h. The reaction solution was cooled to 30℃and poured into 300mL of water, and allowed to stand for 10 hours, whereby a solid was precipitated. Filtering, washing the residue with 100mL saturated sodium bicarbonate aqueous solution to neutrality, and oven drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 100:1, thus obtaining an intermediate (4 l).
4) Methyl 2- ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy) acetate (5 l)
Intermediate (4 l) (1.00 g,2.75 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.165 g,4.125 mmol), and the mixture was reacted at 30℃for 3 minutes, followed by methyl bromoacetate (0.84 g,5.5 mmol) and the reaction was carried out at 30℃for 3 hours. The reaction solution was poured into 200mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (5 l).
5）N-hydroxy-2- ((2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-5-yl) oxy) acetamide (6 l)
0.50g of the intermediate (5 l) was added to 5mL of potassium hydroxylamine methanol solution, reacted at 30℃for 0.3 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 7, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days, and the objective compound (6 l) was obtained.
Preparation of the compounds (6 a) - (6 k) and (6 m) - (6 u), e.g.preparation of (6 l), except for R used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of (6 l).
Example 10
2,2' - ((2- (3-iodophenyl) -4-oxo-4) HBenzopyran-5, 7-diyl) bis (oxy))NThe preparation of hydroxyacetamide) (12 b), as shown in fig. 2, proceeds as follows:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b)
3-iodobenzoic acid (59.52g,240.00 mmol) and 2,4, 6-trihydroxyacetophenone (7) (14.88 g,80.00 mmol) were dissolved in 744mL of pyridine. Phosphorus oxychloride (36.83 g,240.00 mmol) was added dropwise at-5℃for 20 minutes, the ice bath was removed and the reaction was continued for 16 hours at 20 ℃. After the completion of the reaction, the reaction solution was poured into 300mL of water and allowed to stand for 12 hours. Suction filtering, washing the filter cake with water for 2-3 times. Drying at 45-50 ℃ to obtain an intermediate (8 b).
2) Preparation of 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b)
Potassium tert-butoxide (13.42 g,119.85 mmol) was dissolved in 70mLN,NDimethylformamide, nitrogen protection, dropwise addition of intermediate (8 b) (34.28 g,39.95 mmol)N,N114.27mL of dimethylformamide solution was reacted at 20℃for 2 hours. The pH was adjusted to neutral with phosphoric acid, the reaction solution was poured into 300mL of water, and extracted 3 times with 80mL each time of ethyl acetate. The organic phases were combined and washed 3 times with 80mL of saturated aqueous sodium bicarbonate solution and 3 times with 70mL of saturated aqueous sodium chloride solution. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 5:1, so as to obtain an intermediate (9 b).
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b)
Intermediate (9 b) (3.97 g,9.97 mmol) was added to 39.7 mL glacial acetic acid, heated to 80 ℃, 198.5 μl of concentrated sulfuric acid was added, the temperature was raised to 90 ℃, and the reaction was carried out for 2 hours. The reaction solution was cooled to 20℃and poured into 300mL of water, and allowed to stand for 10 hours, whereby a solid was precipitated. Filtering, washing the residue with 100mL saturated sodium bicarbonate aqueous solution to neutrality, and oven drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 100:1, so as to obtain an intermediate (10 b).
4) Dimethyl 2,2' - ((2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-5, 7-diyl) bis (oxy)) diacetate (11 b)
Intermediate (10 b) (2.00 g,5.26 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.42 g10.52 mmol) was reacted at 20℃for 3.5 minutes, methyl bromoacetate (1.61 g,10.52 mmol) was added and reacted at 20℃for 12 hours. The reaction solution was poured into 200mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (11 b).
5) 2,2' - ((2- (3-iodophenyl) -4-oxo-4) HBenzopyran-5, 7-diyl) bis (oxy))NPreparation of-hydroxyacetamide) (12 b)
Intermediate (11 b) (0.70 g,1.33 mmol) was added to 5mL of potassium hydroxylamine in methanol, reacted at 20℃for 1.5 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 6, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days, to give the objective compound (12 b).
Preparation of Compound (12 a), e.g. preparation of Compound (12 b), except R used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (12 b).
Example 11
2,2' - ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5, 7-diyl) bis (oxy))NThe preparation of hydroxyacetamide) (12 b), as shown in fig. 2, proceeds as follows:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b)
3-iodobenzoic acid (65.47g,264.00 mmol) and 2,4, 6-trihydroxyacetophenone (7) (14.88 g,80.00 mmol) were dissolved in 149mL pyridine. Phosphorus oxychloride (40.51 g,264.00 mmol) was added dropwise at 5℃for 20 minutes, the ice bath was removed and the reaction was continued for 2 hours at 40 ℃. After the completion of the reaction, the reaction solution was poured into 300mL of water and allowed to stand for 12 hours. Suction filtering, washing the filter cake with water for 2-3 times. Drying at 45-50 ℃ to obtain an intermediate (8 b).
2) Preparation of 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b)
Potassium tert-butoxide (20.13 g,179.78 mmol) was dissolved in 70mLN,NDimethylformamide, nitrogen protection, dropwise addition of intermediate(8b) (34.28 g,39.95 mmol)N,N68.56mL of dimethylformamide solution and reacted at 40℃for 0.5 hour. The pH was adjusted to neutral with phosphoric acid, the reaction solution was poured into 300mL of water, and extracted 3 times with 80mL each time of ethyl acetate. The organic phases were combined and washed 3 times with 80mL of saturated aqueous sodium bicarbonate solution and 3 times with 70mL of saturated aqueous sodium chloride solution. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to the ethyl acetate is 5:1, so as to obtain an intermediate (9 b).
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b)
Intermediate (9 b) (3.97 g,9.97 mmol) was added to 7.94 mL glacial acetic acid, heated to 90 ℃, 158.8 μl of concentrated sulfuric acid was added, the temperature was raised to 120 ℃, and reacted for 0.5 hours. The reaction solution was cooled to 30℃and poured into 300mL of water, and allowed to stand for 10 hours, whereby a solid was precipitated. Filtering, washing the residue with 100mL saturated sodium bicarbonate aqueous solution to neutrality, and oven drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 100:1, so as to obtain an intermediate (10 b).
4) Dimethyl 2,2' - ((2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-5, 7-diyl) bis (oxy)) diacetate (11 b)
Intermediate (10 b) (2.00 g,5.26 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.63 g,15.78 mmol), and the mixture was reacted at 30℃for 3 minutes, followed by methyl bromoacetate (3.22 g,21.04 mmol) and the reaction was carried out at 30℃for 3 hours. The reaction solution was poured into 200mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (11 b).
5) 2,2' - ((2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5, 7-diyl) bis (oxy))NPreparation of-hydroxyacetamide) (12 b)
Intermediate (11 b) (0.70 g,1.33 mmol) was added to 5mL of potassium hydroxylamine in methanol, reacted at 0℃for 0.3 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 7, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days, to give the objective compound (12 b).
Preparation of Compound (12 a), e.g. preparation of Compound (12 b), except R used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (12 b).
Example 12
N-hydroxy-2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)H-benzopyran-5-yl) oxy) acetamide (15 b), as shown in fig. 2, by the following procedure:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b) was performed as in example 10.
2) 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b) was prepared as in example 10.
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b) was carried out as in example 10.
4) 5-hydroxy-2- (3-iodophenyl) -7-methoxy-4HPreparation of benzopyran-4-one (13 b)
Intermediate (10 b) (0.80 g,2.11 mmol) was dissolved in 100mLN,NTo dimethylformamide was added dimethyl sulfate (0.27 g,2.11 mmol) and potassium carbonate (0.29 g,2.11 mmol). The reaction was carried out at 50℃for 5 hours. The reaction solution was poured into 100mL of water. Standing for 10h. Filtering. Drying at 45-50deg.C. 10mL ethyl acetate was slurried for 20 minutes. Filtering. Intermediate (13 b) is obtained.
5) Methyl 2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)H-benzopyran-5-yl) oxy) acetate (14 b) preparation
Intermediate (13 b) (0.40 g,1.02 mmol) was dissolved in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.04 g,1.02 mmol), and the mixture was reacted at 20℃for 3.5 minutes, followed by addition of methyl bromoacetate (0.156 g,1.02 mmol) and the reaction was carried out at 20℃for 12 hours. The reaction solution was poured into 50mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (14 b).
6）N-hydroxy-2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)HPreparation of benzopyran-5-yl) oxy) acetamide (15 b)
Intermediate (14 b) (0.10 g,0.21 mmol) was added to 2mL of potassium hydroxylamine in methanol, reacted at 20℃for 1.5 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 6, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days, to give the objective compound (15 b).
Preparation of Compounds (15 a), (15 c) as described for the preparation of Compound (15 b), except that R is used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (15 b).
Example 13
N-hydroxy-2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)H-benzopyran-5-yl) oxy) acetamide (15 b), as shown in fig. 2, by the following procedure:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b) was performed as in example 11.
2) 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b) was prepared as in example 11.
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b) was carried out as in example 11.
4) 5-hydroxy-2- (3-iodophenyl) -7-methoxy-4HPreparation of benzopyran-4-one (13 b)
Intermediate (10 b) (0.80 g,2.11 mmol) was dissolved in 100mLN,NTo dimethylformamide, dimethyl sulfate (0.405 g,3.165 mmol) and potassium carbonate (0.435 g,3.165 mmol) were added. The reaction was carried out at 80℃for 1h. The reaction solution was poured into 100mL of water. Standing for 10h. Filtering. Drying at 45-50deg.C. 10mL ethyl acetate was slurried for 20 minutes. Filtering. Intermediate (13 b) is obtained.
5) Methyl 2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)H-benzopyran-5-yl) oxy) acetate (14 b) preparation
Intermediate (13 b) (0.40 g,1.02 mmol) in 10mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.06 g,1.53 mmol), and the mixture was reacted at 30℃for 3 minutes, followed by methyl bromoacetate (0.312 g,2.04 mmol) and the reaction was carried out at 30℃for 3 hours. The reaction solution was poured into 50mL of water and allowed to stand for 10 hours. Filtering, and drying the filter cake at 45-50 ℃. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio of the mobile phase to the methylene dichloride-methanol is 60:1, so as to obtain an intermediate (14 b).
6）N-hydroxy-2- ((2- (3-iodophenyl) -7-methoxy-4-oxo-4)HPreparation of benzopyran-5-yl) oxy) acetamide (15 b)
(0.10 g,0.21 mmol) of the intermediate (14 b) was added to 2mL of a potassium hydroxylamine methanol solution, reacted at 30℃for 0.3 hours, the reaction solution was poured into water, the volume fraction of 10% phosphoric acid was adjusted to pH 7, a white solid was produced, left to stand for 10 hours, filtered and naturally dried for 2 days to give the objective compound (15 b).
Preparation of Compounds (15 a), (15 c) as described for the preparation of Compound (15 b), except that R is used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (15 b).
Example 14
2- ((7- (2-aminoethoxy) -2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NThe preparation of hydroxyacetamide trifluoroacetate salt (19 b), as shown in fig. 2, comprises the following steps:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b) was performed as in example 10.
2) 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b) was prepared as in example 10.
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b) was carried out as in example 10.
4) Tert-butyl (2- ((5-hydroxy-2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-7-yl) oxy) ethyl) carbamate (16 b)
Intermediate (10 b) (1.00 g,2.63 mmol) was dissolved in 100mLN,N-dimethylformamideIn, add intoNBoc-2-bromoethylamine (0.587 g,2.63 mmol), potassium carbonate (0.36 g,2.63 mmol), potassium iodide (0.13 g,0.789 mmol) and tetrabutylammonium bromide (0.25 g,0.789 mmol). The reaction was carried out at 50℃for 5 hours. The reaction solution was poured into 100mL of water. Standing for 10 hours. Filtering. Drying at 45-50deg.C. Column chromatography, mobile phase: dichloromethane-tetrahydrofuran-methanol, the volume ratio of the three is 100:40:0.7, and the intermediate (16 b) is obtained.
5) Methyl 2- ((7- (2-tert-Butoxycarbonyl) amino) ethoxy) -2- (3-iodophenyl) -4-oxo-4H-benzopyran-5-yl) oxy) acetate (17 b) preparation
Intermediate (16 b) (1.20 g,2.29 mmol) was dissolved in 24mLN,NTo dimethylformamide was added 60% by mass of sodium hydride (0.092 g,2.29 mmol), and the mixture was reacted at 20℃for 3.5 minutes, followed by addition of methyl bromoacetate (0.35 g,2.29 mmol) and the reaction was carried out at 20℃for 12 hours. The reaction solution was poured into 50mL of water and allowed to stand for 10 hours. Ethyl acetate was extracted 3 times, 50mL each. The organic phases are combined. 100mL of saturated aqueous sodium chloride solution was washed 1 time. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 70:1, so as to obtain an intermediate (17 b).
6) Tert-butyl (2- ((5- (2- (hydroxyamino) -2-oxoethoxy) -2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-7-yl) oxy) ethyl) carbamate (18 b)
Intermediate (17 b) (0.38 g,0.64 mmol) was added to 5mL of potassium hydroxylamine in methanol, reacted at 20℃for 1.5 hours, and the reaction mixture was poured into 50mL of water. Adjusting pH to 6 with 5% citric acid aqueous solution to obtain white solid, standing for 10 hr, filtering, and naturally drying for 2 days to obtain intermediate (18 b).
7) 2- ((7- (2-aminoethoxy) -2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NPreparation of hydroxyacetamide trifluoroacetate salt (19 b)
Intermediate (18 b) (0.15 g,0.25 mmol) was added to 5mL of dichloromethane followed by 5mL of trifluoroacetic acid. The reaction was carried out at 20℃for 3 hours. The solvent was concentrated off, 10mL ethyl acetate was added and filtered. Naturally drying for 2 days to obtain the target compound (19 b).
Preparation of Compound (19 a) as described for the preparation of Compound (19 b), except that R is used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (19 b).
Example 15
2- ((7- (2-aminoethoxy) -2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NThe preparation of hydroxyacetamide trifluoroacetate salt (19 b), as shown in fig. 2, comprises the following steps:
1) Preparation of 2-acetylphenyl-1, 3, 5-triphenyltris (3-iodobenzoate) (8 b) was performed as in example 11.
2) 1- (3-iodophenyl) -3- (2, 4, 6-trihydroxyphenyl) propane-1, 3-dione (9 b) was prepared as in example 11.
3) 5, 7-dihydroxy-2- (3-iodophenyl) -4HPreparation of benzopyran-4-one (10 b) was carried out as in example 11.
4) Tert-butyl (2- ((5-hydroxy-2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-7-yl) oxy) ethyl) carbamate (16 b)
Intermediate (10 b) (1.00 g,2.63 mmol) was dissolved in 10mLN,N-dimethylformamide, added toNBoc-2-bromoethylamine (0.88 g,3.95 mmol), potassium carbonate (0.72 g,5.26 mmol), potassium iodide (0.87 g,2.63 mmol) and tetrabutylammonium bromide (0.83 g,2.63 mmol). The reaction was carried out at 80℃for 1 hour. The reaction solution was poured into 100mL of water. Standing for 10 hours. Filtering. Drying at 45-50deg.C. Column chromatography, mobile phase: dichloromethane-tetrahydrofuran-methanol, the volume ratio of the three is 100:40:0.7, and the intermediate (16 b) is obtained.
5) Methyl 2- ((7- (2-tert-Butoxycarbonyl) amino) ethoxy) -2- (3-iodophenyl) -4-oxo-4H-benzopyran-5-yl) oxy) acetate (17 b) preparation
Intermediate (16 b) (1.20 g,2.29 mmol) was dissolved in 6mLN,NTo dimethylformamide, 60% sodium hydride (0.138 g,3.44 mmol) was added and reacted at 30℃for 3 minutes, followed by methyl bromoacetateEsters (0.53 g,3.44 mmol) were reacted at 30℃for 3 hours. The reaction solution was poured into 50mL of water and allowed to stand for 10 hours. Ethyl acetate was extracted 3 times, 50mL each. The organic phases are combined. 100mL of saturated aqueous sodium chloride solution was washed 1 time. Dried over anhydrous magnesium sulfate for 15 minutes. Filtering and concentrating. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 70:1, so as to obtain an intermediate (17 b).
6) Tert-butyl (2- ((5- (2- (hydroxyamino) -2-oxoethoxy) -2- (3-iodophenyl) -4-oxo-4)HPreparation of benzopyran-7-yl) oxy) ethyl) carbamate (18 b)
Intermediate (17 b) (0.38 g,0.64 mmol) was added to 5mL of potassium hydroxylamine in methanol, reacted at 30℃for 0.3 hours, and the reaction mixture was poured into 50mL of water. Adjusting pH to 7 with 5% citric acid aqueous solution to obtain white solid, standing for 10 hr, filtering, and naturally drying for 2 days to obtain intermediate (18 b).
7) 2- ((7- (2-aminoethoxy) -2- (3-iodophenyl) -4-oxo-4)HBenzopyran-5-yl) oxy-NPreparation of hydroxyacetamide trifluoroacetate salt (19 b)
Intermediate (18 b) (0.15 g,0.25 mmol) was added to 1.25mL of dichloromethane followed by 2.5mL of trifluoroacetic acid. The reaction was carried out at 30℃for 0.5 hour. The solvent was concentrated off, 10mL ethyl acetate was added and filtered. Naturally drying for 2 days to obtain the target compound (19 b).
Preparation of Compound (19 a) as described for the preparation of Compound (19 b), except that R is used 1 COOH is different; other reaction conditions (e.g., molar amounts of reactants, etc.) and steps are consistent with the preparation of compound (19 b).
Example 16
N-hydroxy-2 ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4) H-benzopyran-5-yl) oxy) acetamide (22 a) as shown in fig. 3, by the following procedure:
1) 5-hydroxy-7- (2-morpholinoethoxy) -2-phenyl-4HPreparation of benzopyran-4-one (20 a)
By varying R used 1 COOH was benzoic acid, obtained according to the preparation method in example 10An intermediate (10 a). Intermediate (10 a) (1.50 g,5.91 mmol) was dissolved in 150mLN,NDimethylformamide, 4- (2-bromoethyl) morpholine hydrobromide (1.63 g,5.91 mmol), potassium carbonate (1.63 g,11.8 mmol), potassium iodide (0.29 g,1.77 mmol) and tetrabutylammonium bromide (0.38 g,1.18 mmol) were added. The reaction was carried out at 50℃for 5 hours. The reaction solution was poured into 100mL of water and allowed to stand for 6 hours. Filtering. Drying at 45-50deg.C to obtain intermediate (20 a).
2) Methyl 2- ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetate (21 a)
Intermediate (20 a) (1.20 g,3.27 mmol) was dissolved in 24mLN,NTo dimethylformamide, 60% sodium hydride (0.13 g,3.27 mmol) was added in portions and reacted at 20℃for 3.5 minutes. Methyl bromoacetate (0.50 g,3.27 mmol) was added. The reaction was carried out at 20℃for 12 hours. Pour into 100mL of water. Ethyl acetate was extracted 3 times, 50mL each. The organic phases are combined. And (5) drying the anhydrous magnesium sulfate. Filtering. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 50:1, thus obtaining the intermediate (21 a).
3）N-hydroxy-2 ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetamide (22 a)
Intermediate (21 a) (0.50 g,1.14 mmol) was added to 8mL of potassium hydroxylamine in methanol. The reaction was carried out at 20℃for 1.5 hours. The reaction was poured into 50mL of water. And adjusting the pH of the phosphoric acid aqueous solution with the volume fraction of 2% to 6-7. Standing for 10 hours. Filtering. Naturally drying to obtain the compound (22 a).
Example 17
N-hydroxy-2 ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetamide (22 a) as shown in fig. 3, by the following procedure:
1) 5-hydroxy-7- (2-morpholinoethoxy) -2-phenyl-4HPreparation of benzopyran-4-one (20 a)
By varying R used 1 COOH was benzoic acid, and intermediate (10 a) was obtained by reference to the preparation method in example 11. Intermediate (10 a) (1.50 g,5.91 mmol) was dissolved in15mLN,NDimethylformamide, 4- (2-bromoethyl) morpholine hydrobromide (2.44 g,8.86 mmol), potassium carbonate (2.44 g,17.7 mmol), potassium iodide (0.97 g,5.91 mmol) and tetrabutylammonium bromide (0.95 g,2.95 mmol) were added. The reaction was carried out at 90℃for 1 hour. The reaction solution was poured into 100mL of water and allowed to stand for 6 hours. Filtering. Drying at 45-50deg.C to obtain intermediate (20 a).
2) Methyl 2- ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetate (21 a)
Intermediate (20 a) (1.20 g,3.27 mmol) was dissolved in 6mLN,NTo dimethylformamide, sodium hydride (0.195 g,4.90 mmol) was added in portions and reacted at 30℃for 3 minutes. Methyl bromoacetate (0.75 g,4.90 mmol) was added. The reaction was carried out at 30℃for 3 hours. Pour into 100mL of water. Ethyl acetate was extracted 3 times, 50mL each. The organic phases are combined. And (5) drying the anhydrous magnesium sulfate. Filtering. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 50:1, thus obtaining the intermediate (21 a).
3）N-hydroxy-2 ((7- (2-morpholinoethoxy) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetamide (22 a)
Intermediate (21 a) (0.50 g,1.14 mmol) was added to 8mL of potassium hydroxylamine in methanol. The reaction was carried out at 30℃for 0.3 hour. The reaction was poured into 50mL of water. And adjusting the pH of the phosphoric acid aqueous solution with the volume fraction of 2% to 6-7. Standing for 10 hours. Filtering. Naturally drying to obtain the compound (22 a).
Example 18
N-hydroxy-2- ((7- (2-morpholin-2-oxyethyl) -4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetamide (27 a) as shown in fig. 3, by the following procedure:
1) Methyl 2- ((5-hydroxy-4-oxo-2-phenyl-4)HBenzopyran-7-yl) oxo) acetate (23 a)
By varying R used 1 COOH was benzoic acid, and intermediate (10 a) was obtained by reference to the preparation method in example 10. Intermediate (10 a) (4.00 g,15.75 mmol) was dissolved in 40mLN,NAdding potassium carbonate to dimethylformamide(2.17 g,15.75 mmol) and methyl bromoacetate (3.47 g,15.75 mmol). The reaction was carried out at 40℃for 5 hours. The reaction solution was poured into 200mL of water. Standing for 5 hours. Filtering. Drying at 45-50 ℃ to obtain an intermediate (23 a).
2) 2- ((5-hydroxy-4-oxo-2-phenyl-4)H-benzopyran-7-yl) oxy) acetic acid (24 a) preparation
Intermediate (23 a) (4.90 g,15.03 mmol) was added to 49mL of methanol, and potassium hydroxide (1.68 g,30.06 mmol) was added. The reaction was carried out at 50℃for 2 hours. The reaction solution was poured into 200mL of water, and the pH was adjusted to be neutral with a 10% volume fraction of phosphoric acid aqueous solution. Standing for 1 hour. Filtering. Drying at 45-50 ℃ to obtain an intermediate (24 a).
3) 5-hydroxy-7- (2-morpholin-2-yloxyethoxy) -2-phenyl-4HPreparation of benzopyran-4-one (25 a)
Intermediate (24 a) (1.50 g,4.81 mmol) was dissolved in 15mLN,N-dimethylformamide. 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (0.92 g,4.81 mmol), 1-hydroxybenzotriazole (0.65 g,4.81 mmol) and 4-dimethylaminopyridine (0.059 g,0.48 mmol) were added at-5 ℃. Reaction was carried out at-5℃for 30 min, and morpholine (0.42 g,4.81 mmol) was added. The reaction was carried out at 20℃for 12 hours. The reaction solution was poured into 100mL of water and allowed to stand for 8 hours. Filtering. Drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 200:1, so as to obtain an intermediate (25 a).
4) Methyl 2- ((7- (2-morpholin-2-yloxyethoxy) -4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetate (26 a) preparation
Intermediate (25 a) (0.72 g,1.89 mmol) was dissolved in 3.6mLN,NTo dimethylformamide, 60% sodium hydride (75.53 mg,1.89 mmol) was added in portions. After 3.5 minutes of reaction at 25℃methyl bromoacetate (0.29 g,1.89 mmol) was added. After reacting at 20℃for 12 hours, the reaction mixture was poured into 100mL of water. Ethyl acetate was extracted 3 times, 50mL each. The organic phases were combined and dried over anhydrous magnesium sulfate. Filtering. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio is 30:1, thus obtaining an intermediate (26 a).
5）N-hydroxy-2- (. About.7- (2-morpholin-2-yloxyethyl) -4-oxo-2-phenyl-4HPreparation of benzopyran-5-yl) oxy) acetamide (27 a)
Intermediate (26 a) (0.50 g,1.10 mmol) was added to 8mL of potassium hydroxylamine in methanol and reacted at 20℃for 1.5 hours. After the reaction is finished, concentrating the methanol; adding water, adjusting pH to 6-7 with 10% phosphoric acid aqueous solution, standing for 12 hr, filtering, and naturally drying to obtain compound (27 a).
Example 19
N-hydroxy-2- ((7- (2-morpholin-2-oxyethyl) -4-oxo-2-phenyl-4) H-benzopyran-5-yl) oxy) acetamide (27 a) as shown in fig. 3, by the following procedure:
1) Methyl 2- ((5-hydroxy-4-oxo-2-phenyl-4)HBenzopyran-7-yl) oxo) acetate (23 a)
By varying R used 1 COOH was benzoic acid, and intermediate (10 a) was obtained by reference to the preparation method in example 11. Intermediate (10 a) (4.00 g,15.75 mmol) was dissolved in 400mLN,NTo dimethylformamide was added potassium carbonate (4.34 g,31.50 mmol) and methyl bromoacetate (5.21 g,23.62 mmol). The reaction was carried out at 80℃for 0.5 hour. The reaction solution was poured into 200mL of water. Standing for 5 hours. Filtering. Drying at 45-50 ℃ to obtain an intermediate (23 a).
2) 2- ((5-hydroxy-4-oxo-2-phenyl-4)H-benzopyran-7-yl) oxy) acetic acid (24 a) preparation
Intermediate (23 a) (4.90 g,15.03 mmol) was added to 490mL of methanol and potassium hydroxide (6.64 g,120.24 mmol) was added. The reaction was carried out at 65℃for 0.5 hour. The reaction solution was poured into 200mL of water, and the pH was adjusted to be neutral with a 10% volume fraction of phosphoric acid aqueous solution. Standing for 1 hour. Filtering. Drying at 45-50 ℃ to obtain an intermediate (24 a).
3) 5-hydroxy-7- (2-morpholin-2-yloxyethoxy) -2-phenyl-4HPreparation of benzopyran-4-one (25 a)
Intermediate (24 a) (1.50 g,4.81 mmol) was dissolved in 150mLN,N-dimethylformamide. 1-Ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.38 g,7.21 mmol), 1-hydroxy were added at 5 ℃Benzotriazole (0.97 g,7.21 mmol) and 4-dimethylaminopyridine (0.295 g,2.80 mmol). Morpholine (0.63 g,7.21 mmol) was added after 30 minutes at 5 ℃. The reaction was carried out at 25℃for 6 hours. The reaction solution was poured into 100mL of water and allowed to stand for 8 hours. Filtering. Drying at 45-50deg.C. Purifying by column chromatography, wherein the mobile phase is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol is 200:1, so as to obtain an intermediate (25 a).
4) Methyl 2- ((7- (2-morpholin-2-yloxyethoxy) -4-oxo-2-phenyl-4)H-benzopyran-5-yl) oxy) acetate (26 a) preparation
Intermediate (25 a) (0.72 g,1.89 mmol) was dissolved in 14.4mLN,NTo dimethylformamide, 60% sodium hydride (113.30 mg,2.83 mmol) was added in portions. After 3 minutes of reaction at 30℃methyl bromoacetate (0.58 g,3.78 mmol) was added. After 3 hours of reaction at 30 ℃, the reaction solution was poured into 100mL of water. Ethyl acetate was extracted 3 times, 50mL each. The organic phases were combined and dried over anhydrous magnesium sulfate. Filtering. Purifying by column chromatography, wherein the mobile phase is methylene dichloride-methanol, and the volume ratio is 30:1, thus obtaining an intermediate (26 a).
5）N-hydroxy-2- ((7- (2-morpholin-2-oxyethyl) -4-oxo-2-phenyl-4)HPreparation of benzopyran-5-yl) oxy) acetamide (27 a)
Intermediate (26 a) (0.50 g,1.10 mmol) was added to 8mL of potassium hydroxylamine in methanol and reacted at 30℃for 0.3 hours. After the reaction is finished, concentrating the methanol; adding water, adjusting pH to 6-7 with 10% phosphoric acid aqueous solution, standing for 12 hr, filtering, and naturally drying to obtain compound (27).

Claims (9)

1. A flavone derivative aminopeptidase N inhibitor, which is characterized by having a structure represented by the formula (I):
wherein:
R 1 is that
R 2 Is H, methoxy,
2. A process for the preparation of a flavone derivative aminopeptidase N inhibitor as defined in claim 1, wherein when R 2 Is H, methoxy,In the process, the intermediate shown in the formula (II) reacts with methyl bromoacetate in the presence of sodium hydride to generate an intermediate shown in the formula (III), the intermediate shown in the formula (III) is hydrolyzed in a potassium hydroxylamine methanol solution, and then acidized to generate the flavone derivative aminopeptidase N inhibitor;
wherein: r is R 3 Is H, hydroxy, methoxy,R 4 Is H,Methoxy group,/->
3. The process of claim 2, wherein when R 3 In the case of methoxy, the intermediate shown in the formula (II) is obtained by reacting the intermediate shown in the formula (VII) with dimethyl sulfate under the alkaline condition of potassium carbonate;
4. the process of claim 2, wherein when R 3 Is thatIn the process, an intermediate shown in a formula (II) is obtained by reacting an intermediate shown in a formula (VII) with 4- (2-bromoethyl) morpholine hydrobromide under the condition of potash property;
5. the process of claim 2, wherein when R 3 Is thatThe intermediate shown in the formula (II) reacts with methyl bromoacetate under the alkaline condition of potassium carbonate, and then is hydrolyzed under the alkaline condition and condensed with morpholine to obtain the intermediate shown in the formula (VII);
6. the process of claim 2, wherein when R 3 In the case of H, 2, 6-dihydroxyacetophenone and carboxyl groupAcid R 1 COOH reaction to form an intermediate of formula (X); under alkaline conditions, the intermediate shown in the formula (X) undergoes Baker-Venkataraman rearrangement to generate the intermediate shown in the formula (XI); in a glacial acetic acid/concentrated sulfuric acid system, performing cyclization reaction on an intermediate shown in a formula (XI) to generate an intermediate shown in a formula (II);
7. a process for the preparation of a flavone derivative aminopeptidase N inhibitor as defined in claim 1, wherein when R 2 Is thatReacting an intermediate shown in a formula (IV) with methyl bromoacetate in the presence of sodium hydride to generate an intermediate shown in a formula (V), hydrolyzing the intermediate shown in the formula (V) in a potassium hydroxylamine methanol solution, then acidifying to generate an intermediate shown in a formula (VI), and removing tert-butoxycarbonyl groups from the intermediate shown in the formula (VI) under the acidic condition of trifluoroacetic acid to generate the flavone derivative aminopeptidase N inhibitor; an intermediate shown in a formula (IV) is obtained by reacting an intermediate shown in a formula (VII) with N-tert-butoxycarbonyl-2-bromoethylamine under a potash condition;
8. the process according to any one of claims 2 to 6, wherein when R 3 In the case of hydroxyl, the intermediate represented by the formula (II) or the intermediate represented by the formula (VII) is prepared by the following preparation method:
2,4, 6-Trihydroxyacetophenone and carboxylic acid R 1 COOH reaction to form an intermediate of formula (VIII); under alkaline conditions, an intermediate of formula (VIII)The body undergoes Baker-Venkataraman rearrangement to produce an intermediate of formula (IX); in a glacial acetic acid/concentrated sulfuric acid system, performing cyclization reaction on an intermediate shown in a formula (IX) to generate an intermediate shown in a formula (II) or a formula (VII);
9. A pharmaceutical composition suitable for oral or parenteral administration comprising a flavone derivative aminopeptidase N inhibitor, or a pharmaceutically acceptable salt thereof, according to claim 1 and one or more pharmaceutically acceptable carriers or excipients.
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Title
曹江营.1,2,3-三氮唑酰胺类氨肽酶N抑制剂设计合成和活性筛选.中国新药杂志.2022,第31卷第1922-1928页. *

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