阅读说明：本技术 一种可作为gpr35受体激动剂的苯并吡喃酮衍生物、制备方法及其应用 (Benzopyrone derivative capable of being used as GPR35 receptor agonist, preparation method and application thereof ) 是由 梁鑫淼 魏来 赵耀鹏 王纪霞 程俊翔 侯滔 王俊 于 2020-03-18 设计创作，主要内容包括：本发明提供了一种可作为GPR35受体激动剂的苯并吡喃酮衍生物、制备方法及其应用。该类化合物对人源GPR35受体普遍表现出激动活性,是人源性GPR35受体的特异性激动剂。本发明提供的化合物是GPR35受体的活性配体,该类化合物及其药学上可接受的盐、溶剂化物、水合物或晶型对人源GPR35受体普遍表现出较高的激动活性。本发明是GPR35受体的特异性激动剂,可应用于治疗、预防和抑制与GPR35受体介导相关的疾病。(The invention provides a benzopyrone derivative capable of being used as a GPR35 receptor agonist, and a preparation method and application thereof. The compounds generally show agonistic activity on human GPR35 receptor, and are specific agonists of human GPR35 receptor. The compound provided by the invention is an active ligand of a GPR35 receptor, and the compound and pharmaceutically acceptable salts, solvates, hydrates or crystal forms thereof generally show higher agonistic activity on a human GPR35 receptor. The invention is a specific agonist of GPR35 receptor, and can be applied to treating, preventing and inhibiting diseases related to GPR35 receptor mediation.)

1. Benzopyrone derivatives serving as GPR35 receptor agonists, which comprise pharmaceutically acceptable salts and have the structural general formula shown as the following:

wherein the content of the first and second substances,

ar is a substituted or unsubstituted aryl or heterocyclic aryl group including phenyl, thienyl, furyl, pyridyl, or naphthyl;

R₁independently selected from nitro, methoxy, hydroxy, halogen;

R₂independently selected from cyano, 1H-tetrazolyl.

2. The benzopyrone derivatives as claimed in claim 1, which are agonists of the GPR35 receptor and characterized in that: ar is optionally substituted with one or more substituents: -H, -C_1-4Alkyl, -OC_1-4Alkoxy, benzyloxy, phenoxymethylene, -COOC_1-4Alkoxyformyl, ethoxyformyl, carboxy, N-dimethyl, -CN, halogen, -CF₃、-NO₂and-OH is substituted at any position.

3. The benzopyrone derivatives as claimed in claim 1, which are agonists of the GPR35 receptor and characterized in that: ar is optionally substituted with one or more substituents: -H, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, benzyloxy, phenoxymethylene, ethoxyformyl, carboxy, N-dimethyl, -CN, -F, -Cl, -Br, -I, -CF₃、-NO₂and-OH is substituted at any position.

4. The benzopyrone derivatives as claimed in claim 1, which are agonists of the GPR35 receptor and characterized in that: ar is substituted or unsubstituted phenyl, thienyl, furyl, wherein Ar is optionally substituted with one or more substituents: methyl, ethyl, isopropyl, methoxy, ethoxy, phenoxymethylene, ethoxyformyl, carboxyl, N-dimethyl and-F are substituted at any position.

5. The benzopyrone derivatives as claimed in claim 1, which are agonists of the GPR35 receptor and characterized in that: r₁Is nitro.

6. The benzopyrone derivatives as claimed in claim 1, which are agonists of the GPR35 receptor and characterized in that: r₂Is 1H-tetrazolyl.

7. The benzopyrone derivatives as claimed in claim 1, which are agonists of the GPR35 receptor and characterized in that: when Ar is thienyl, the site of attachment to the benzopyrone nucleus may be the 2-or 3-position on the thiophene ring; when Ar is furyl, the site of attachment to the benzopyrone nucleus may be at the 2-or 3-position on the furan ring.

8. The benzopyrone derivatives as claimed in claim 1, which are agonists of the GPR35 receptor and characterized in that: wherein the derivative is preferably selected from the group consisting of compounds (1) to (31):

(1)6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(2)6- (3-fluorophenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(3)6- (3-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(4)6- (4-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(5)6- (4-fluorophenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(6) 8-Nitro-2-oxo-6- (thien-2-yl) -2H-chromene-3-carbonitrile

(7)6- (4-ethylphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(8)6- (4-ethylphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(9)6- (2-fluoro-4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(10)6- (2-fluoro-4-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(11)6- (2-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(12)6- (2-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(13)6- (4-ethoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(14)6- (4-ethoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(15)6- (3-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(16)6- (4-isopropylphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(17)6- (4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(18)6- (4-isopropylphenyl) -8-methoxy-2-oxo-2H-chromene-3-carbonitrile

(19) 8-methoxy-6- (4-methoxyphenyl) -2-oxo-2H-chromene-3-carbonitrile

(20)6- (4-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(21)6- (4-isopropylphenyl) -8-methoxy-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(22)6- (4- (benzyloxy) phenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(23)6- (4- (benzyloxy) phenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(24)4- (3-cyano-8-nitro-2-oxo-2H-chromen-6-yl) benzoic acid ethyl ester

(25)4- (8-Nitro-2-oxo-3- (1H-tetrazol-5-yl) -2H-chromen-6-yl) benzoic acid ethyl ester

(26)3- (8-Nitro-2-oxo-3- (1H-tetrazol-5-yl) -2H-chromen-6-yl) benzoic acid ethyl ester

(27)6- (4- (dimethylamino) phenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(28) 8-Nitro-2-oxo-6- (p-tolyl) -2H-chromene-3-carbonitrile

(29) 8-Nitro-3- (1H-tetrazol-5-yl) -6- (p-tolyl) -2H-pyran-2-one

(30)6- (furan-2-yl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(31)6- (furan-2-yl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one.

9. The benzopyrone derivatives as GPR35 receptor agonists according to any one of claims 1 to 8, wherein the pharmaceutically acceptable salts of the derivatives comprise lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, ammonium salt, and one or more of pharmaceutically acceptable solvates, hydrates or crystal forms thereof.

10. A process for the preparation of benzopyrone derivatives useful as agonists of the GPR35 receptor, which process is carried out by a reaction scheme as described in the following formula:

reaction conditions are as follows: (a) arylboronic acids, PdCl₂(PPh₃)₂,Na₂CO₃,DMF:H₂O＝1:1,80℃,4h；(b)NH₄Cl,H₂O,8h.(c)HCl(2M),75℃,1h.(d)AlCl3,NaN₃THF,80 ℃, overnight;

the reaction scheme involves the following steps: firstly, under proper conditions, carrying out suzuki coupling on a salicylaldehyde derivative A and a boric acid derivative B to obtain a product C; under appropriate conditions, C is reacted with malononitrile to give R₂Target product D being a cyano group; and finally, reacting the D with sodium azide under proper conditions to obtain a target product E. Wherein, Ar and R₁、R₂As defined in any one of claims 1 to 8 above.

11. The application of benzopyrone derivatives serving as GPR35 receptor agonists in preparing medicaments for treating, preventing and relieving diseases regulated by GPR35 receptor active ligands is characterized in that the derivatives also comprise pharmaceutically acceptable salts, solvates, hydrates or various crystal forms and pharmaceutical compositions thereof.

12. The use of claim 11, wherein the disease comprises one or more of colon cancer, stomach cancer, inflammation, allergy, pain, inflammatory bowel disease, asthma, enteritis, coronary artery disease, hypertension, parkinson's disease, and non-alcoholic fatty liver.

13. The use according to claim 11, wherein the medicament comprises a pharmaceutical composition comprising one or more than two of the benzopyrone derivatives and pharmaceutically acceptable salts thereof according to any one of claims 1 to 8, and any pharmaceutically acceptable excipients, or other active compounds.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a benzopyrone derivative capable of serving as a GPR35 receptor agonist, and a preparation method and application thereof.

Background

G Protein Coupled Receptors (GPCRs) are the most successful drug receptors in the history of drug development, and 30-40% of drugs used in modern clinic use G protein coupled receptors as targets (Trends in GPCR drug delivery: new agents, targets and indicators. nature reviews drug delivery 2017, 16: 829-. There are several GPCR ligands among the best-selling 100 drugs in the world. However, only 108 receptors have been developed as drug targets in all G protein-coupled receptor families. Therefore, such targets still have great development potential in drug development (Target-based drug discovery: is somenting wrongDrug discovery. today.2005,10, 139-.

The G protein-coupled receptor 35(GPR35) is an orphan receptor first discovered in 1998 (GPR35 is a functional receptor in a multiple root growth receptors, biochem. Biophys. Res. Commun.2008,365, 344-348). Studies to date have shown that this receptor is associated with a number of diseases. However, physiological studies of this receptor have been hampered by the lack of appropriate pharmacological tools and the absence of endogenous ligands. Although some endogenous molecules such as kynurenic acid activate the GPR35 receptor, they are not considered endogenous ligands due to their generally low activity. Therefore, the search for highly potent ligands for this receptor has helped to investigate the physiological significance of this receptor in vivo.

Modern pharmacological studies indicate that GPR35 may be involved in the treatment of a variety of diseases, such as cancer, inflammation, coronary artery disease, allergy, pain, asthma and hypertension (Discovery of 2- (4-Methylfuran-2(5H) -ylene) malononitriles and Thieno 3,2-b thiolene-2-carboxylic Acid Derivatives as G Protein-Coupled Receptor 35(GPR35) antibodies.j.med.chem.2011, 54,7385 @ 7396.). Among the drugs that have been marketed, the antiasthmatic drugs, cromolyn sodium and minoxidil (Zaprinast, a well-known cyclic guanosine monophosphate-specific phosphoric acid synthase inhibitor, is an agonist for GPR35 receptor 35.febs lett.2006,580, 5003-5008). These findings suggest that our GPR35 receptor holds promise as a drug receptor. The development of novel, highly potent agonists of the GPR35 receptor is of great interest.

Benzopyrone compounds are lactone compounds widely existing in nature and mostly exist in rutaceae and umbelliferae plants, and the anticoagulant drug, namely the bis-benzopyrone, has been proved to be a GPR35 receptor agonist.

Disclosure of Invention

In view of the foregoing, it is a first object of the present invention to provide a benzopyrone derivative, including pharmaceutically acceptable salts, solvates, hydrates or crystal forms thereof, which is useful as GPR35 receptor agonist and has the following structural formula:

wherein the content of the first and second substances,

ar is a substituted or unsubstituted aryl or heterocyclic aryl group including phenyl, thienyl, furyl, pyridyl, or naphthyl;

R₁independently selected from nitro, methoxy, hydroxy, halogen;

R₂independently selected from cyano, 1H-tetrazolyl.

Preferably, Ar is optionally substituted with one or more substituents: -H, -C_1-4Alkyl, -OC_1-4Alkoxy, benzyloxy, phenoxymethylene, -COOC_1-4Alkoxyformyl, ethoxyformyl, carboxy, N-dimethyl, -CN, halogen, -CF₃、-NO₂and-OH is substituted at any position.

Further preferred, Ar is optionally substituted with one or more substituents: -H, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, benzyloxy, phenoxymethylene, ethoxyformyl, carboxy, N-dimethyl, -CN, -F, -Cl, -Br, -I, -CF₃、-NO₂and-OH is substituted at any position.

Second preferred, Ar is substituted or unsubstituted phenyl, thienyl, furyl, wherein Ar is optionally substituted with one or more substituents: methyl, ethyl, isopropyl, methoxy, ethoxy, phenoxymethylene, ethoxyformyl, carboxyl, N-dimethyl and-F are substituted at any position.

Preferably, R₁Is nitro.

Preferably, R₂Is 1H-tetrazolyl.

Preferably, when Ar is thienyl, the site of attachment to the benzopyrone nucleus may be the 2-or 3-position on the thiophene ring; when Ar is furyl, the site of attachment to the benzopyrone nucleus may be at the 2-or 3-position on the furan ring.

In some embodiments, wherein the derivative is preferably selected from compounds (1) - (31):

(1)6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(2)6- (3-fluorophenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(3)6- (3-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(4)6- (4-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(5)6- (4-fluorophenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(6) 8-Nitro-2-oxo-6- (thien-2-yl) -2H-chromene-3-carbonitrile

(7)6- (4-ethylphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(8)6- (4-ethylphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(9)6- (2-fluoro-4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(10)6- (2-fluoro-4-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(11)6- (2-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(12)6- (2-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(13)6- (4-ethoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(14)6- (4-ethoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(15)6- (3-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(16)6- (4-isopropylphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(17)6- (4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(18)6- (4-isopropylphenyl) -8-methoxy-2-oxo-2H-chromene-3-carbonitrile

(19) 8-methoxy-6- (4-methoxyphenyl) -2-oxo-2H-chromene-3-carbonitrile

(20)6- (4-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(21)6- (4-isopropylphenyl) -8-methoxy-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(22)6- (4- (benzyloxy) phenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(23)6- (4- (benzyloxy) phenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(24)4- (3-cyano-8-nitro-2-oxo-2H-chromen-6-yl) benzoic acid ethyl ester

(25)4- (8-Nitro-2-oxo-3- (1H-tetrazol-5-yl) -2H-chromen-6-yl) benzoic acid ethyl ester

(26)3- (8-Nitro-2-oxo-3- (1H-tetrazol-5-yl) -2H-chromen-6-yl) benzoic acid ethyl ester

(27)6- (4- (dimethylamino) phenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

(28) 8-Nitro-2-oxo-6- (p-tolyl) -2H-chromene-3-carbonitrile

(29) 8-Nitro-3- (1H-tetrazol-5-yl) -6- (p-tolyl) -2H-pyran-2-one

(30)6- (furan-2-yl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

(31)6- (furan-2-yl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

And pharmaceutically acceptable salts, solvates, hydrates or various crystal forms of the compounds.

Further, pharmaceutically acceptable salts of the above compounds, especially lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, and ammonium salt.

A second object of the present invention is to provide a process for the preparation of benzopyrone derivatives useful as agonists of the GPR35 receptor, which can be prepared by the reaction scheme described by the following formula:

reaction conditions are as follows: (a) arylboronic acids, PdCl₂(PPh₃)₂,Na₂CO₃,DMF:H₂O＝1:1,80℃,4h。(b)NH₄Cl,H₂O,8h.(c)HCl(2M),75℃,1h.(d)AlCl₃,NaN₃THF,80 ℃ overnight.

The reaction scheme involves the following steps:

(a) under proper conditions, carrying out suzuki coupling on a salicylaldehyde derivative A and a boric acid derivative B to obtain a product C;

(b) under appropriate conditions, C is reacted with malononitrile to give R₂Target product D being a cyano group;

(c) and (3) reacting the D with sodium azide under proper conditions to obtain a target product E, wherein Ar, R1 and R2 are the same as the definitions.

The benzopyrone derivatives and pharmaceutically acceptable salts thereof disclosed in the present invention can be easily prepared by skilled researchers based on the conditions disclosed in the present invention. Based on the synthesis conditions disclosed in the invention, the reaction scale is enlarged or reduced in an equal proportion, or the amount of the solvent used or the reaction temperature or the reaction time is changed within a reasonable range, or the equivalent of substances such as a reaction substrate, a catalyst, an additive and the like is changed within a reasonable range, which belongs to the protection scope of the invention.

The third purpose of the invention is to provide a benzopyrone derivative which can be used as a GPR35 receptor agonist, and comprises pharmaceutically acceptable salts, solvates, hydrates or pharmaceutical compositions of various crystal forms thereof, and can further comprise excipients, diluents and carriers. The compounds of the invention may exist in undissolved and solvated forms with pharmaceutically acceptable solvents. The solvated forms are generally considered equivalent to unsolvated forms. The pharmaceutical compositions of the present invention may include one or more compounds of the present invention, and typical formulations are prepared by mixing a compound of the present invention, or a pharmaceutically acceptable salt, solvate, hydrate thereof, with a carrier, excipient, or diluent. Common carriers, excipients or diluents include substances such as carbohydrates, cellulose and its derivatives, gelatin, oils, polyols, water, etc. The dosage form of the medicine is solid preparation or liquid preparation, and specifically is tablet, capsule, powder, granule, solution, syrup, suspension or aerosol.

The fourth purpose of the invention is to provide an application of benzopyrone derivatives which can be used as GPR35 receptor agonists in preparing medicines for treating, preventing and relieving diseases modulated by GPR35 receptor active ligands, wherein the derivatives comprise pharmaceutically acceptable salts, solvates, hydrates or various crystal forms thereof. Such diseases include cancer, inflammation, coronary artery disease, allergy, pain, inflammatory bowel disease, asthma and hypertension which may be modulated by agonists and partial agonists of the GPR35 receptor. The medicament is a GPR35 receptor agonist. The medicine comprises one or more than two of benzopyrone derivatives and pharmaceutically acceptable salts thereof, and a pharmaceutical composition consisting of any pharmaceutically acceptable auxiliary materials or other active compounds.

The invention also provides a synthetic method of the benzopyrone derivative, and a series of benzopyrone derivatives taking benzopyrone as a parent nucleus are synthesized. The series of compounds were tested for activity at the human GPR35 receptor by acting on the human GPR35 receptor. The activity of inflammatory bowel disease is tested by acting one of the compounds on mice, and the potential of the compound for treating the inflammatory bowel disease is determined. The results prove that the series of compounds have certain drug development prospect.

Drawings

FIG. 1: the general structural formula of the derivative is shown in the figure;

FIG. 2: the preparation method of the derivative of the invention is a reaction scheme;

FIG. 3: a is the dose-dependent DMR signature for compound 20; b is the dose-dependent curve of the compound;

FIG. 4: evaluation of pharmacological activity of compound 20 in mice;

Detailed Description

The compounds of the general formula of the present invention can be prepared by the following reaction scheme.

Reaction conditions are as follows: (a) arylboronic acids, PdCl₂(PPh₃)₂,Na₂CO₃,DMF:H₂O＝1:1,80℃,4h。(b)NH₄Cl,H₂O,8h.(c)HCl(2M),75℃,1h.(d)AlCl₃,NaN₃THF,80 ℃ overnight. The reaction scheme involves the following steps: firstly, under proper conditions, carrying out suzuki coupling on a salicylaldehyde derivative A and a boric acid derivative B to obtain a product C; under appropriate conditions, C is reacted with malononitrile to give R₂Target product D being a cyano group; and finally, reacting the D with sodium azide under proper conditions to obtain a target product E.

The starting materials employed in the preparation of the compounds of the invention are known, can be prepared according to known methods or are commercially available.

The following various preparative procedures were used to prepare synthetic intermediates for the compounds of the present invention.

Preparation of 1, 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

1.0mmol of the starting material 5-bromo-2-hydroxy-3-nitrobenzaldehyde was dissolved in 20mL of DMF displaced with nitrogen: h₂To O (V: V ═ 1: 1), then to the solution were added 0.05 equivalents of bis (triphenylphosphine) palladium dichloride, 6.0 equivalents of sodium carbonate, and 1.5 equivalents of 3-fluorobenzeneboronic acid in one portion at room temperature. Stirring evenly after the addition is finished, replacing the residual air in the system with nitrogen for 2 times, and then heating to 80 ℃ under the protection of nitrogen and stirring for reaction for 4 hours. After the reaction is finished, adding excessive concentrated hydrochloric acid into the solution, filtering out solid after the product is fully separated out, dissolving the solid by using ethanol, filtering out the solid, passing through diatomite, and performing rotary evaporation to obtain a solid product which can be directly used for the next reaction.

Preparation of 2, 3 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 3-methoxybenzeneboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 3, 4 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 4-fluorobenzeneboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 4, 2-hydroxy-3-nitro-5- (thien-2-yl) benzaldehyde

And (3) replacing the 3-fluorobenzene boric acid in the preparation 1 with thiophene-2-boric acid, and obtaining a target intermediate product under the same conditions.

Preparation of 5, 4 '-ethyl-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzene boric acid in the preparation 1 with thiophene-2-boric acid, and obtaining a target intermediate product under the same conditions.

Preparation of 6, 2' -fluoro-4-hydroxy-4 ' -methoxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 2-fluoro-4-methoxybenzeneboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 7, 2 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 2-methoxybenzeneboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 8, 4 '-ethoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 4-ethoxyphenylboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 9, 3 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 3-methoxybenzeneboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 10, 4 '-isopropyl-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzene boric acid in the preparation 1 with 4-isopropylbenzene boric acid, and obtaining a target intermediate product under the same conditions.

Preparation of 11, 4 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 4-methoxybenzeneboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 12, 4-hydroxy-4 '-isopropyl-5-methoxy- [1,1' -biphenyl ] -3-carbaldehyde

Replacing 5-bromo-2-hydroxy-3-nitrobenzaldehyde in the preparation 1 with 5-bromo-2-hydroxy-3-methoxybenzaldehyde, replacing 3-fluorobenzeneboronic acid with 4-isopropylphenylboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 13, 4-hydroxy-4 ', 5-dimethoxy- [1,1' -biphenyl ] -3-carbaldehyde

Replacing 5-bromo-2-hydroxy-3-nitrobenzaldehyde in the preparation 1 with 5-bromo-2-hydroxy-3-methoxybenzaldehyde, replacing 3-fluorobenzeneboronic acid with 4-methoxybenzeneboronic acid, and obtaining the target intermediate product under the same conditions.

Preparation of 14, 4'- (benzyloxy) -4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 4-benzyloxy phenylboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 15, 3 '-formyl-4' -hydroxy-5 '-nitro- [1,1' -biphenyl ] -4-carboxylic acid ethyl ester

The 3-fluorobenzeneboronic acid in the preparation 1 is changed into (4- (ethoxycarbonyl) phenyl) boronic acid, and the target intermediate product can be obtained under the same conditions.

Preparation of 16, 3 '-formyl-4' -hydroxy-5 '-nitro- [1,1' -biphenyl ] -3-carboxylic acid ethyl ester

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with (3- (ethoxycarbonyl) phenyl) boronic acid, and obtaining a target intermediate product under the same condition.

Preparation of 17, 4'- (dimethylamino) -4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

The 3-fluorobenzeneboronic acid in the preparation 1 is changed into (4- (dimethylamino) phenyl) boronic acid, and the target intermediate product can be obtained under the same conditions.

Preparation of 18, 4-hydroxy-4 '-methyl-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with 4-methylbenzeneboronic acid, and obtaining a target intermediate product under the same conditions.

Preparation of 19, 5- (furan-2-yl) -2-hydroxy-3-nitrobenzaldehyde

And (3) replacing the 3-fluorobenzeneboronic acid in the preparation 1 with furan-2-boronic acid, and obtaining a target intermediate product under the same conditions.

Example 1: 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

1.0mmol of 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-formaldehyde is uniformly dispersed in 20mL of water, then 1.0 equivalent of ammonium acetate is added into the system, after uniform stirring, 2.0 equivalents of malononitrile is added into the solution, then stirring reaction is carried out for 8h at normal temperature, after the reaction is finished, solid is filtered out, the obtained solid is added into dilute hydrochloric acid (2M) and stirring reaction is carried out for 1h at 75 ℃. And (3) filtering out a solid after the reaction is finished, dissolving the obtained solid by dichloromethane, adding a proper amount of silica gel, and performing spin drying to pass through a column by pure dichloromethane to obtain a pure target product, wherein the product is a yellow solid, and the yield is 76%.

¹H NMR(400MHz,DMSO-d₆)δ8.99(s,1H),8.75(d,J＝2.1Hz,1H),8.52(d,J＝2.1Hz,1H),7.73(d,J＝10.4Hz,1H),7.68(d,J＝7.9Hz,1H),7.62(dd,J＝14.0,7.8Hz,1H),7.35(t,J＝8.2Hz,1H)。

Example 2: 6- (3-fluorophenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

1.0mmol of 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile was dissolved in 20mL of ultra-dry tetrahydrofuran, then 3.0 equivalents of aluminum chloride and 6.0 equivalents of sodium azide were added to the solution, and the mixture was heated to 80 ℃ under nitrogen protection and stirred at reflux overnight. After completion of the reaction, an excess of dilute hydrochloric acid (2M) was added to the solution and the solution was extracted 2 times with ethyl acetate. The organic phases were combined, the organic phase was dried over anhydrous sodium sulfate and spin dried, then the solid was washed 2 times with a small amount of dichloromethane (2mL × 2) and 2 times with a small amount of methanol (2mL × 2) to obtain the final product as a yellow solid in 57% yield.

¹H NMR(400MHz,DMSO-d₆)δ9.12(s,1H),8.78(s,1H),8.71(d,J＝2.0Hz,1H),7.74(dd,J＝14.9,9.2Hz,2H),7.63(dd,J＝14.4,7.8Hz,1H),7.34(t,J＝8.2Hz,1H).

Example 3: 6- (3-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was obtained in the same procedure as in example 1 except for using 3 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde instead of 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1, and the product was obtained as a yellow solid with a yield of 64%.

¹H NMR(400MHz,DMSO-d₆)δ9.00(s,1H),8.71(d,J＝2.2Hz,1H),8.48(d,J＝2.2Hz,1H),7.48(t,J＝8.1Hz,1H),7.37(d,J＝6.5Hz,2H),7.11-7.02(m,1H),3.86(s,3H).

Example 4: 6- (4-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was obtained in the same procedure as in example 1 except for using 4 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde instead of 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1, and the product was a yellow solid with a yield of 79%.

¹H NMR(400MHz,DMSO-d₆)δ9.00(s,1H),8.69(d,J＝2.2Hz,1H),8.45(d,J＝2.2Hz,1H),7.87(dd,J＝8.7,5.3Hz,2H),7.42(t,J＝8.8Hz,2H).

Example 5: 6- (4-fluorophenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 58% yield by the same procedure of example 2, substituting 6- (4-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.15(s,1H),8.72(d,J＝2.1Hz,1H),8.66(d,J＝2.2Hz,1H),7.91(dd,J＝8.8,5.3Hz,2H),7.42(t,J＝8.8Hz,2H).

Example 6: 8-Nitro-2-oxo-6- (thien-2-yl) -2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 71% yield using the same procedure as in example 1, substituting 2-hydroxy-3-nitro-5- (thiophen-2-yl) benzaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.02(s,1H),8.69(d,J＝2.3Hz,1H),8.39(d,J＝2.2Hz,1H),7.76(dd,J＝5.9,4.4Hz,2H),7.24(dd,J＝5.0,3.7Hz,1H).

Example 7: 6- (4-ethylphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 81% yield using the same procedure as in example 1, substituting 4 '-ethyl-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.02(s,1H),8.67(s,1H),8.45(s,1H),7.74(d,J＝7.9Hz,2H),7.41(d,J＝7.9Hz,2H),2.69(q,J＝7.4Hz,2H),1.22(d,J＝7.6Hz,3H).

Example 8: 6- (4-ethylphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 58% yield using the same procedure of example 2, substituting 6- (4-ethylphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.15(s,1H),8.72(d,J＝2.2Hz,1H),8.63(d,J＝2.2Hz,1H),7.77(d,J＝8.2Hz,2H),7.41(d,J＝8.2Hz,2H),2.69(q,J＝7.5Hz,2H),1.23(d,J＝7.6Hz,3H).

Example 9: 6- (2-fluoro-4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 65% yield using the same procedure as in example 1, substituting 2' -fluoro-4-hydroxy-4 ' -methoxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3' -fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.05(s,1H),8.49(d,J＝2.2Hz,1H),8.25(d,J＝2.2Hz,1H),7.50(dd,J＝8.5,6.8Hz,1H),7.15(dd,J＝11.4,2.4Hz,1H),6.97(td,J＝8.4,2.5Hz,1H),3.85(s,3H).

Example 10: 6- (2-fluoro-4-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 77% yield using the same procedure of example 2, substituting 6- (2-fluoro-4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.16(s,1H),8.50(d,J＝2.1Hz,1H),8.46(d,J＝2.1Hz,1H),7.52(dd,J＝8.4,6.8Hz,1H),7.15(dd,J＝11.4,2.4Hz,1H),6.97(td,J＝8.4,2.4Hz,1H),3.87(s,3H).

Example 11: 6- (2-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 78% yield by the same procedure of example 1, substituting 2 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.03–8.95(m,1H),8.71(d,J＝2.2Hz,1H),8.48(d,J＝2.2Hz,1H),7.48(t,J＝8.1Hz,1H),7.40–7.28(m,2H),7.07(dd,J＝7.3,2.1Hz,1H),3.90–3.80(m,3H).

Example 12: 6- (2-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 65% yield using the same procedure of example 2, substituting 6- (2-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.18(s,1H),8.52(d,J＝2.1Hz,1H),8.48(d,J＝2.1Hz,1H),7.47(ddd,J＝7.4,6.0,1.7Hz,2H),7.22(d,J＝8.2Hz,1H),7.13(dd,J＝11.2,3.7Hz,1H),3.85(s,3H).

Example 13: 6- (4-ethoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 78% yield by the same procedure of example 1, substituting 4 '-ethoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.00(s,1H),8.64(d,J＝2.3Hz,1H),8.41(d,J＝2.3Hz,1H),7.78–7.70(m,2H),7.10(d,J＝8.8Hz,2H),4.11(q,J＝7.0Hz,2H),1.37(t,J＝7.0Hz,3H).

Example 14: 6- (4-ethoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 71% yield using the same procedure of example 2, substituting 6- (4-ethoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.14(s,1H),8.68(d,J＝2.2Hz,1H),8.60(d,J＝2.2Hz,1H),7.78(d,J＝8.8Hz,2H),7.10(d,J＝8.8Hz,2H),4.11(q,J＝6.9Hz,2H),1.37(t,J＝7.0Hz,3H).

Example 15: 6- (3-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 72% yield by the same procedure of example 2, substituting 3 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.12(s,1H),8.74(d,J＝1.9Hz,1H),8.66(d,J＝1.6Hz,1H),7.48(t,J＝7.8Hz,1H),7.41(d,J＝7.3Hz,2H),7.09–7.03(m,1H),3.87(s,3H).

Example 16: 6- (4-isopropylphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 81% yield using the same procedure as in example 1, substituting 4 '-isopropyl-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.02(s,1H),8.67(s,1H),8.44(s,1H),7.74(d,J＝7.7Hz,2H),7.44(d,J＝7.8Hz,2H),2.98(dt,J＝13.8,7.0Hz,1H),1.25(d,J＝6.9Hz,6H).

Example 17: 6- (4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 65% yield by the same procedure of example 1, substituting 4 '-methoxy-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.00(s,1H),8.64(d,J＝2.3Hz,1H),8.41(d,J＝2.3Hz,1H),7.79–7.73(m,2H),7.12(d,J＝8.8Hz,2H),3.84(s,3H).

Example 18: 6- (4-isopropylphenyl) -8-methoxy-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 87% yield by the same procedure of example 1, substituting 4-hydroxy-4 '-isopropyl-5-methoxy- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ8.95(s,1H),7.68(dd,J＝10.1,4.9Hz,3H),7.61(d,J＝1.6Hz,1H),7.39(d,J＝8.2Hz,2H),4.04(s,3H),2.96(dt,J＝13.7,6.8Hz,1H),1.25(d,J＝6.9Hz,6H).

Example 19: 8-methoxy-6- (4-methoxyphenyl) -2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 87% yield by the same procedure of example 1, substituting 4-hydroxy-4 ', 5-dimethoxy- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ8.93(s,1H),7.70(d,J＝8.7Hz,2H),7.67(d,J＝1.8Hz,1H),7.58(d,J＝1.7Hz,1H),7.08(d,J＝8.7Hz,2H),4.04(s,3H),3.82(s,3H).

Example 20: 6- (4-methoxyphenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 82% yield using the same procedure of example 2, substituting 6- (4-methoxyphenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.14(s,1H),8.68(d,J＝2.1Hz,1H),8.60(d,J＝2.2Hz,1H),7.80(d,J＝8.8Hz,2H),7.13(d,J＝8.8Hz,2H),3.84(s,3H).

Example 21: 6- (4-isopropylphenyl) -8-methoxy-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 82% yield using the same procedure of example 2, substituting 6- (4-isopropylphenyl) -8-methoxy-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.06(s,1H),7.85(d,J＝1.9Hz,1H),7.70(d,J＝8.2Hz,2H),7.65(d,J＝1.8Hz,1H),7.39(d,J＝8.2Hz,2H),4.73(s,1H),4.06(s,3H),2.96(dt,J＝13.8,6.9Hz,1H),1.26(d,J＝6.9Hz,6H).

Example 22: 6- (4- (benzyloxy) phenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 65% yield using the same procedure as example 1, substituting 4'- (benzyloxy) -4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ8.99(s,1H),8.64(d,J＝2.3Hz,1H),8.42(d,J＝2.2Hz,1H),7.76(d,J＝8.8Hz,2H),7.49(d,J＝7.1Hz,2H),7.42(t,J＝7.3Hz,2H),7.35(t,J＝7.2Hz,1H),7.20(d,J＝8.8Hz,2H),5.20(s,2H).

Example 23: 6- (4- (benzyloxy) phenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 78% yield using the same procedure of example 2, substituting 6- (4- (benzyloxy) phenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.14(s,1H),8.68(s,1H),8.61(d,J＝2.1Hz,1H),7.80(d,J＝8.7Hz,2H),7.49(d,J＝7.3Hz,2H),7.42(t,J＝7.3Hz,2H),7.36(d,J＝7.0Hz,1H),7.21(d,J＝8.7Hz,2H),5.21(s,2H).

Example 24: 4- (3-cyano-8-nitro-2-oxo-2H-chromen-6-yl) benzoic acid ethyl ester

This compound was prepared as a yellow solid in 74% yield by the same procedure of example 1, substituting 4 '-formyl-4' -hydroxy-5 '-nitro- [1,1' -biphenyl ] -4-carboxylic acid ethyl ester for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.03(s,1H),8.78(d,J＝2.2Hz,1H),8.55(d,J＝2.2Hz,1H),8.12(d,J＝8.4Hz,2H),7.98(d,J＝8.4Hz,2H),4.37(q,J＝7.1Hz,2H),1.36(t,J＝7.1Hz,3H).

Example 25: 4- (8-Nitro-2-oxo-3- (1H-tetrazol-5-yl) -2H-chromen-6-yl) benzoic acid ethyl ester

This compound was prepared as a yellow solid in 45% yield using the same procedure of example 2, substituting 4- (3-cyano-8-nitro-2-oxo-2H-chromen-6-yl) benzoic acid ethyl ester for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromen-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.16(s,1H),8.82(s,1H),8.74(s,1H),8.13(d,J＝8.2Hz,2H),8.01(d,J＝8.2Hz,2H),4.37(q,J＝7.0Hz,2H),1.36(t,J＝7.1Hz,3H).

Example 26: 3- (8-Nitro-2-oxo-3- (1H-tetrazol-5-yl) -2H-chromen-6-yl) benzoic acid ethyl ester

This compound was prepared as a yellow solid in 68% yield using the same procedure of example 2, substituting 3- (3-cyano-8-nitro-2-oxo-2H-chromen-6-yl) benzoic acid ethyl ester for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromen-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.20(s,1H),8.81(d,J＝1.8Hz,1H),8.72(d,J＝2.0Hz,1H),8.36(s,1H),8.13(d,J＝7.8Hz,1H),8.07(d,J＝7.8Hz,1H),7.73(t,J＝7.8Hz,1H),4.39(q,J＝7.1Hz,2H),1.37(t,J＝7.1Hz,3H).

Example 27: 6- (4- (dimethylamino) phenyl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 36% yield using the same procedure of example 2, substituting 6- (4- (dimethylamino) phenyl) -8-nitro-2-oxo-2H-methylene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.05(s,1H),8.59(d,J＝2.2Hz,1H),8.51(d,J＝2.2Hz,1H),7.69(d,J＝8.8Hz,2H),6.86(d,J＝8.9Hz,2H),2.98(s,6H).

Example 28: 8-Nitro-2-oxo-6- (p-tolyl) -2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 87% yield by the same procedure of example 1, substituting 4-hydroxy-4 '-methyl-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.01(s,1H),8.67(d,J＝2.3Hz,1H),8.45(d,J＝2.3Hz,1H),7.72(d,J＝8.2Hz,2H),7.38(d,J＝8.0Hz,2H),2.39(s,3H).

Example 29: 8-Nitro-3- (1H-tetrazol-5-yl) -6- (p-tolyl) -2H-pyran-2-one

This compound was prepared as a yellow solid in 78% yield using the same procedure of example 2, substituting 8-nitro-2-oxo-6- (p-tolyl) -2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.10(s,1H),8.70(d,J＝2.1Hz,1H),8.61(d,J＝2.2Hz,1H),7.75(d,J＝8.1Hz,2H),7.38(d,J＝8.0Hz,2H),2.39(s,3H).

Example 30: 6- (furan-2-yl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile

This compound was prepared as a yellow solid in 69% yield using the same procedure of example 1, substituting 5- (furan-2-yl) -2-hydroxy-3-nitrobenzaldehyde for 3 '-fluoro-4-hydroxy-5-nitro- [1,1' -biphenyl ] -3-carbaldehyde in example 1.

¹H NMR(400MHz,DMSO-d₆)δ9.05(s,1H),8.71(d,J＝2.2Hz,1H),8.41(d,J＝2.1Hz,1H),7.92(d,J＝1.2Hz,1H),7.28(d,J＝3.4Hz,1H),6.72(dd,J＝3.5,1.8Hz,1H).

Example 31: 6- (furan-2-yl) -8-nitro-3- (1H-tetrazol-5-yl) -2H-chromen-2-one

This compound was prepared as a yellow solid in 67% yield using the same procedure of example 2, substituting 6- (furan-2-yl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile for 6- (3-fluorophenyl) -8-nitro-2-oxo-2H-chromene-3-carbonitrile of example 2.

¹H NMR(400MHz,DMSO-d₆)δ9.13(s,1H),8.65(d,J＝5.2Hz,2H),7.92(s,1H),7.24(d,J＝3.0Hz,1H),6.72(s,1H).

Pharmacological characterization-DMR screening model

The material HT-29 cells are purchased from Shanghai cell bank of Chinese academy of sciences; minoxidil was purchased from Sigma. The detection platform is the third generation of kangningAn imager, the detected signal being a wavelength shift caused by a cell Dynamic Mass Reset (DMR).

HT-29 cells in logarithmic growth phase were seeded in different wells of a 384-well plate at a seeding volume of 40. mu.L per well, and the number of cells seeded per well was 3.2X 10⁴And (3) placing the inoculated cell plate in a cell culture box for culturing for 20-22h until the cell fusion degree reaches about 95%.

All compounds described above were then tested separately for gradient DMR activity on this cell line, with the activity results shown in the table below.

TABLE 1 Activity data for compounds of the general formula in DMR screening models.

In this example, the term "alkyl" denotes a straight-chain or branched alkyl-C group of 1 to 4 carbon atoms_1-4. The hydrocarbyl group may be selected from methyl, ethyl, propyl, butyl, and isomers thereof. For example, propyl includes n-propyl and isopropyl, and butyl includes isobutyl, sec-butyl, tert-butyl, and the like.

The term "halogen" denotes-F, -Cl, -Br and-I.

The term "alkoxy" denotes a straight or branched alkyl-O-C group having 1 to 4 carbon atoms, bonded via an oxygen atom_1-4. The following are examples that may be mentioned: methoxy, ethoxy, propoxy, butoxy, and isomers thereof.

Animal model test of IBD:

25 males, 8 weeks, with a body weight of 22-26g, were divided into 6 mice in the blank group, 6 mice in the Dextran Sodium Sulfate (DSS) group, 6 mice in the mesalazine + DSS control group, and 6 mice in the compound 20+ DSS administration group. The building block was dosed daily with drinking water (5% DSS); the control group was administered with water (5% DSS) daily and mesalazine was administered by intragastric administration (5 mg/kg); the compound groups were administered daily with water (5% DSS) and compound 20 was administered by intraperitoneal injection (0.2mg/10 g). After one week of feeding, the colon is taken, and the length of the colon of each group is compared, so that the compound 20 is obviously increased in the length of the colon compared with that of a DSS induced control group, which indicates that the compound has a certain IBD treatment effect, and also indicates that the compounds with the structures have a certain IBD treatment effect.

The above description is only a preferred embodiment of the present patent and does not limit the present patent in any way. It will be understood by those skilled in the art that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.