CN115504940A

CN115504940A - Amide compound, preparation method and pharmaceutical application thereof

Info

Publication number: CN115504940A
Application number: CN202110699107.XA
Authority: CN
Inventors: 蒋翔锐; 王延轶; 张波; 彭珂; 付玉志; 李锐; 张哲瑞; 张玉兰; 蒋华良; 沈敬山
Original assignee: Shanghai Institute of Materia Medica of CAS; Wuhan Institute of Virology of CAS
Current assignee: Shanghai Institute of Materia Medica of CAS; Wuhan Institute of Virology of CAS
Priority date: 2021-06-23
Filing date: 2021-06-23
Publication date: 2022-12-23
Also published as: WO2022268145A1; CN118076589A

Abstract

The disclosure relates to an amide compound represented by a general formula I, a prodrug, a pharmaceutically acceptable salt, a complex or a solvate thereof. The compound disclosed by the invention has a novel structure and remarkable antiviral activity and anti-inflammatory activity.

Description

Amide compound, preparation method and pharmaceutical application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to an amide compound, and a preparation method and pharmaceutical application thereof.

Background

Niclosamide was approved by the U.S. food and drug administration in 1960 for the treatment of intestinal tapeworm infection, and its action mechanism is to inhibit the oxidative phosphorylation process of mitochondria in somatic cells of the worm, to block the absorption and uptake of glucose, and further to kill parasites. Niclosamide is safe for use for over fifty years by now and is listed in the world health organization's basic drug list. In addition, niclosamide can also be used for treating animal tapeworm infection, killing Oncomelania hupensis Gredler and preventing schistosomiasis.

Recent studies have shown that niclosamide has a wide range of biological activities and can modulate a variety of signaling pathways and biological processes, such as Wnt/β -catenin, mTORC1, STAT3, NF- κ B, notch, NS2B-NS protein interactions, etc. (Cancer Lett,2014,349,8-14 cell res,27, 1046-1064. Furthermore, niclosamide exhibits pharmacological antiviral effects (Tuberculosis, 2019,116, S28-S33; nat. Med,2016,22, 1101-1107.)

Coronaviruses are a widely distributed single-stranded positive-strand RNA virus that infects a variety of higher animals, and are also common human pathogens. For example, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1 often cause mild cold symptoms in humans (Academic Press,2018,100, 163-188). However, coronaviruses such as SARS-CoV, MERS-CoV and SARS-CoV-2 cause serious respiratory diseases and even death in human body. Effective treatment methods for patients infected with coronavirus still remain to be developed. Wu et al found that niclosamide inhibited the replication of SARS-CoV and completely inhibited the synthesis of viral antigen at 1.5. Mu.M by screening old drugs for new drugs on the market (Antimicrob. Agents Chemother,2004,48, 2693-2696.). Salix et al found that niclosamide inhibited the cytopathic effect (CPE) of SARS-CoV at 1. Mu.M and inhibited SARS-CoV replication in Vero E6 cells with an EC50 value of less than 0.1. Mu.M (J.Med.chem, 2007,50, 4087-4095).

Flavivirus is also a single-stranded positive-strand RNA virus, and mainly includes Zika virus (Zika virus), dengue virus (dengue virus), japanese encephalitis virus (Japanese encephalitis virus), west Nile virus (West Nile virus), yellow fever virus (yellow fever virus), and many of these viruses are human pathogens. Niclosamide is known to be a potential inhibitor of zika virus infection by enzyme screening methods, with an intracellular level of IC50 of 0.37 μ M, and is found to directly inhibit flavivirus NS2B-NS3 interactions, while niclosamide is also found to be a broad-spectrum inhibitor of flaviviruses (Cell Res,2017,27,1046-1064 nat. Med,2016,22,1101-1107.

Hepatitis C Virus (HCV) is also an envelope positive single stranded RNA virus of the flaviviridae family. About 7100 million people worldwide suffer from chronic HCV infection, with niclosamide having an EC50 for inhibiting HCV viral replication of 0.16 μ M, and this mechanism of action for inhibiting viral replication is similar to that of nitazoxanide and tizoxanide which regulates host cell progression (antipicrob. Agents chemicother, 2008,52,4069-4071, antipiral res,2011,91,233-240 acs med. Chem.lett,2011,2, 849-854.

Ebola virus is a virulent infectious disease virus that causes ebola hemorrhagic fever in humans and primates. Niclosamide has a strong ebola virus inhibitory activity with an EC50 of 1.5 μ M (ACS infection. Dis,2015,1, 317-326).

Human rhinovirus, a member of the family picornaviridae, rhinovirus, is the leading cause of common cold and presents a serious health risk to patients with asthma, chronic lung disease, and infantile severe bronchiolitis. Niclosamide is a pH-dependent HRV infection inhibitor, with IC50 reaching the micromolar level; it acts as a proton carrier, inhibiting HRV virus entry by blocking acidification of the endolysosomal compartment (clin. Microbiol. Rev,2013,26,135-162 plos pathog,2012 8, e 1002976.

Human adenovirus (Human adenoviruses) is a non-enveloped, double-stranded DNA virus, with seven subspecies in total. The prevalence of adenoviruses in the human population, especially infants and adolescents, is mainly associated with respiratory, ocular-and gastrointestinal infections. Currently, there are no truly viable drugs for treating such viral infections (FEMS microbiol.rev.2019,43, 380-388. The research finds that niclosamide can remarkably inhibit the infection of human adenovirus, and the IC50 is 0.6 mu M.

However, niclosamide has some obvious disadvantages, such as low oral bioavailability, insufficient antiviral activity, etc., which limit the further development and application of niclosamide as an antiviral active compound. On the basis of the structure of niclosamide, the structural optimization is carried out, and the compound with stronger antiviral activity and higher oral bioavailability is found, and the broad-spectrum antiviral active compound with the anti-inflammatory effect has very important significance.

Disclosure of Invention

Object of the Invention

In order to solve the disadvantages of the prior art, it is an object of the present invention to provide a class of amide compounds having antiviral activity, in particular, activity against coronavirus, bunyavirus and dengue virus.

It is an object of the present invention to provide a process for the preparation of the above compounds.

It is a further object of the present invention to provide the use of the above compounds in the manufacture of a medicament for the treatment or prevention of viral infections.

Technical scheme

According to one aspect of the present invention, there is provided an amide compound represented by the following general formula I, a prodrug, a pharmaceutically acceptable salt, a complex, or a solvate thereof:

wherein the content of the first and second substances,

y is a phenyl ring, a 5-6 membered heteroaromatic ring, a phenyl and 5-6 membered heteroaromatic ring or a 5-6 membered heteroaromatic and 5-6 membered heteroaromatic ring; preferably a benzene ring, a pyridine ring or a thiazole ring;

z is O or S;

A ₁ is C-OR ₁₁ 、C-SR ₁₁ 、C-NHR ₁₁ N or CR ₁₂ ；

R ₁₁ Is a hydrogen atom, a C1-C20 alkyl group, a C1-C20 alkylcarbonyl group, a C1-C20 alkylaminocarbonyl group, a C1-C20 alkylcarbonyloxy C1-C20 alkyl group,

C1-C20 alkylphosphoryl, C1-C20 alkylsulfonyl, C1-C20 alkyl-substituted 6-to 20-membered heterocyclylcarbonyl or C1-C20 alkyl-substituted heterocyclylcarbonyl containing 1 to 3 heteroatoms selected from O, N or S

The C1-C20 alkyl group is optionally selected from hydroxyl, amino (-NH) ₂ ) C1-C20 alkylamino, cyano, carboxyl, halogen,

C1-C10 alkyl sulfonyl, C1-C10 alkyl phosphoryl and C1-C10 acyl;

preferably, R ₁₁ Is hydrogen atom, C1-C10 alkyl, C1-C10 alkylcarbonyl, C1-C10 alkylaminocarbonyl, C1-C10 alkylcarbonyloxy, C1-C10 alkylSubstituted 6-to 10-membered heterocyclylcarbonyl or

The above C1-C10 alkyl group is optionally substituted by a substituent selected from the group consisting of a hydroxyl group, an amino group, a cyano group, a carboxyl group, a halogen,

C1-C10 alkyl sulfonyl, C1-C10 alkyl phosphoryl and C1-C10 alkyl carbonyl;

more preferably, R ₁₁ Is a hydrogen atom, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, an aminoC 1-C6 alkyl group, a hydroxyl group, a C1-C6 alkyl group, a C1-C6 alkylcarbonyl group, a C1-C6 alkylaminocarbonyl group, a C1-C6 alkylcarbonyloxy C1-C6 alkyl group, a C1-C6 alkylaminoC 1-C6 alkylcarbonyl group, an aminoC 1-C6 alkylcarbonyl group, a,

Substituted C1-C6 alkylcarbonyl, C1-C6 alkyl-substituted 6-to 8-membered heterocyclylcarbonyl containing 1 to 3 heteroatoms selected from O, N or S or

Most preferably, R ₁₁ Is a hydrogen atom, a C1-C4 alkyl group, a C1-C6 alkylcarbonyl group, a C1-C5 alkylaminocarbonyl group, a C1-C5 alkylcarbonyloxy C1-C5 alkyl group,

Substituted C1-C4 alkyl substituted C1-C4 alkylcarbonyl,

Or

R ₇ Is C1-C10 alkyl, C1-C10 alkylamino C1-C10 alkyl or C3-C10 cycloalkyl, preferably C1-C6 alkyl, C1-C6 alkylaminoC1-C6 alkyl or C3-C7 cycloalkyl, more preferably C1-C4 alkyl, C1-C3 alkylamino C1-C4 alkyl or C3-C6 cycloalkyl;

R ₈ is hydrogen atom, halogen or C1-C10 alkyl; preferably a hydrogen atom, a halogen or a C1-C2 alkyl group, more preferably a hydrogen atom or a methyl group;

R ₁₂ is hydrogen atom, halogen, cyano, nitro or C1-C20 alkyl, preferably hydrogen atom, halogen, cyano, nitro or C1-C6 alkyl; more preferably a hydrogen atom, a halogen, a cyano group or a C1-C4 alkyl group;

A ₂ 、A ₃ 、A ₄ 、A ₅ each independently is N or CR ₂ (ii) a Preferably, A ₂ 、A ₃ 、A ₄ 、A ₅ At most two of N;

R ₂ are the same or different from each other and are each independently selected from hydrogen, hydroxy, halogen, C1-C6 alkyloxy, C1-C6 alkyl, amino (-NH) ₂ ) Nitro, cyano, carboxyl, aldehyde, C1-C6 alkylcarbonyl, C1-C6 alkylsulfonyl, C1-C6 alkylsulfonylamino and C1-C6 alkyloxycarbonyl, said hydroxyl, amino, carboxyl, C1-C6 alkyl being optionally substituted by one or more of hydroxyl, amino, cyano, carboxyl, halogen, C1-C10 alkylsulfonyl, C1-C10 alkylphosphoryl, C1-C10 alkylcarbonyl;

preferably, R ₂ Are the same or different from each other and are each independently selected from the group consisting of hydrogen, hydroxy, halogen, C1-C4 alkyloxy, C1-C4 alkyl, amino, nitro, cyano and C1-C4 alkylcarbonyl, said C1-C4 alkyl being optionally substituted by one or more of hydroxy, amino, cyano, carboxy, halogen, C1-C4 alkylsulfonyl, C1-C4 alkylphosphoryl, C1-C4 alkylcarbonyl;

more preferably, R ₂ Are identical or different from one another and are each independently selected from the group consisting of hydrogen, hydroxy, halogen, C1-C2 alkoxy, C1-C2 alkyl, C1-C2 alkylcarbonyl, haloC 1-C2 alkoxy, haloC 1-C2 alkyl, haloC 1-C2 alkylcarbonyl, amino and nitro;

R ₃ is a hydrogen atom or a C1-C6 alkyl group, preferably R ₃ Is a hydrogen atom or a C1-C4 alkyl group, more preferably R ₃ Is a hydrogen atom;

R ₄ to R ₆ Each independently selected from the group consisting of hydrogen, hydroxy, amino, halogen, nitro, cyano, C1-C6 alkyl, C1-C6 alkyloxy and C1-C6 alkylthio, said C1-C6 alkyl being optionally substituted by one or more of hydroxy, amino, cyano, carboxy, halogen, C1-C6 alkylsulfonyl, C1-C6 alkylphosphoryl, C1-C6 alkylacyl;

preferably, R ₄ To R ₆ Each independently selected from hydrogen atom, hydroxyl, amino, halogen, nitro, cyano, C1-C4 alkyl and C1-C4 alkyloxy, the C1-C4 alkyl is optionally substituted by one or more of hydroxyl, amino, cyano, carboxyl and halogen;

more preferably, R ₄ To R ₆ Each independently selected from a hydrogen atom, a halogen, a C1-C4 alkyl group, a halogenated C1-C4 alkyl group, a C1-C4 alkoxy group, and a halogenated C1-C4 alkoxy group.

In the present invention, halogen means fluorine, chlorine, bromine, iodine; halogenated C1-C4 alkyl means C1-C4 alkyl substituted with one or more halogen atoms, preferably trifluoromethyl; halogenated C1-C4 alkoxy means C1-C4 alkyloxy substituted with one or more halogen atoms, preferably trifluoromethoxy.

In some embodiments, the amide compound represented by formula I is a compound represented by formula II, IIIA or IIIB below:

wherein the content of the first and second substances,

A ₁ is C-OR ₁₁ 、C-SR ₁₁ 、C-NHR ₁₁ Or CR ₁₂ ；

R ₁₁ 、R ₁₂ 、R ₂ 、R ₄ To R ₆ And Y is as defined for formula I.

In some embodiments, the amide compound represented by formula I is a compound represented by formula IV below:

wherein the content of the first and second substances,

R ₁₁ 、R ₂ 、R ₄ and R ₅ Are as defined in formula I.

In some embodiments, the above R ₂ Each independently selected from hydrogen, halogen (especially fluorine and chlorine), hydroxyl, formyl, acetyl, nitro.

In some embodiments, R is as defined above ₄ And R ₅ Each independently selected from the group consisting of hydrogen, halogen, cyano, methyl, trifluoromethyl, methoxy, trifluoromethoxy, methylthio, trifluoromethylthio.

In some embodiments, the above R ₁₁ Is hydrogen.

In some embodiments, the compound or salt is selected from the following structures:

the invention also provides a preparation method of the compound with the general formula.

Wherein, the general formula II, IIIA or IIIB can be prepared by the following method I or method II:

the method comprises the following steps:

preparing compounds II and IIIA with general formulas by condensation reaction, and obtaining condensation products II and IIIA under the condition of a condensing agent or phosphorus trichloride;

the second method comprises the following steps:

taking the condensation product as a raw material, and carrying out alkylation, acylation, phosphorylation or sulfonylation reaction on the compound I and/or IIIA to obtain a compound IIIB.

Wherein each group is as defined above.

In another aspect, the present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the group consisting of the above-described compounds of the present invention, prodrugs, pharmaceutically acceptable salts, complexes and solvates thereof, and optionally a pharmaceutically acceptable carrier.

In another aspect, the present invention provides the use of said compound, a prodrug, a pharmaceutically acceptable salt, complex or solvate thereof, or said pharmaceutical composition, in the manufacture of an antiviral medicament.

In some embodiments, the virus is at least one selected from the group consisting of a DNA virus, an RNA virus.

In some embodiments, the DNA virus is selected from the group consisting of hepatitis b virus, human papilloma virus, herpes virus, phage virus; the RNA virus is selected from influenza virus, avian influenza virus, zika virus, dengue fever virus, respiratory syncytial virus, SARS virus, MERS virus, novel coronavirus (SARS-CoV-2), bunyavirus, AIDS virus, ebola virus, hepatitis C virus, encephalitis B virus, rhinovirus, poliovirus, coxsackie virus, rotavirus, tobacco mosaic virus, bacteriophage virus, marburg virus and enterovirus.

In some embodiments, the RNA virus is at least one of a coronavirus, a bunyavirus, and a dengue virus.

In some embodiments, the coronavirus is SARS-CoV-2.

In some embodiments, the bunyavirus is rift valley fever virus.

In some embodiments, the virus is dengue virus.

In another aspect, the present invention provides the use of said compound, prodrug, pharmaceutically acceptable salt, complex or solvate thereof, or said pharmaceutical composition in the manufacture of a medicament against an inflammatory disease.

In some embodiments, the inflammatory disease is an inflammatory skin disease, an inflammatory bowel disease, an inflammatory joint disease, and other autoimmune diseases.

Advantageous effects

The compound of the invention has the following beneficial effects: novel structure and remarkable antiviral activity.

Detailed Description

For further understanding of the present invention, the present invention is further illustrated by the following examples, which are described only to illustrate the features of the present invention in further detail, and are not intended to limit the scope of the present invention or the scope of the claims of the present invention. The invention is not limited to the above embodiments, but may be modified in various ways.

Wherein the compounds of formula (la) can be prepared by a variety of synthetic methods, a series of synthetic steps, well known to those skilled in the art.

Example 1: synthesis of N- (4-trifluoromethylthiazole) -5-chlorosalicylamide (IIIA-1)

Sequentially adding weighed 2-hydroxy-5-chloro-benzoic acid into an eggplant-shaped bottle, adding toluene (10 times volume) dried by a molecular sieve, adding 2-amino-4-trifluoromethylthiazole with equivalent weight, adding phosphorus trichloride (about 1.1 equivalent weight), immediately replacing with nitrogen, heating to 115 ℃ from room temperature, stirring at the temperature for 8 hours, and gradually clarifying the reaction solution. Detecting aniline by TLC, post-treating, adding a little methanol to quench phosphorus trichloride, concentrating to remove most solvent, extracting with ethyl acetate and water, separating, and saturating with sodium chlorideThe organic layer was washed with solution, dried over anhydrous sodium sulfate, filtered, and sample-mixed, eluting with petroleum ether and ethyl acetate (50: 51.8 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ12.34(s,1H),11.73(s,1H),8.05(d,J＝1.1Hz,1H),7.90(d,J＝2.7Hz,1H),7.52(dd,J＝8.8,2.8Hz,1H),7.08(d,J＝8.8Hz,1H).ESI-MS m/z 307.9(M-H) ^- .

Example 2: synthesis of N- (2-chloro-4-trifluoromethoxyphenyl) -5-chlorosalicylamide (IIIA-2)

IIIA-2 was prepared in the same manner as in example 1 except that 2-chloro-4-trifluoromethoxyaniline was used in place of 2-amino-4-trifluoromethylthiazole and the reaction time was 5 hours, in the following yield: 65.9 percent. ¹ H NMR(600MHz,DMSO-d6)δ12.31(s,1H),10.96(s,1H),8.51(d,J＝9.1Hz,1H),7.98(d,J＝2.8Hz,1H),7.73(d,J＝2.8Hz,1H),7.52(dd,J＝8.7,2.8Hz,1H),7.47(dt,J＝9.1,1.8Hz,1H),7.08(d,J＝8.8Hz,1H).ESI-MS m/z 363.9(M-H) ^- .

Example 3: synthesis of N- (4-trifluoromethylthiazole) -5-chlorosalicylamide (IIIA-3)

IIIA-3 was prepared in the same manner as in example 1 except that 2-amino-5-trifluoromethylthiazole was used instead of 2-amino-4-trifluoromethylthiazole, with the following yields: and (5) percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ12.33(s,2H),8.22(s,1H),7.87(d,J＝2.7Hz,1H),7.52(dd,J＝8.8,2.8Hz,1H),7.06(d,J＝8.8Hz,1H).ESI-MS m/z 320.9(M-H) ^- .

Example 4: synthesis of N- (4-trifluoromethylphenyl) -5-chlorosalicylamide (IIIA-4)

IIIA-4 was prepared in the same manner as in example 1 except that 4-trifluoromethylaniline was used instead of 2-amino-4-trifluoromethylthiazole and the reaction time was 7 hours, in such a yield that: 67.36 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.57(s,1H),10.64(s,1H),7.94(d,J＝8.5Hz,2H),7.88(d,J＝2.7Hz,1H),7.77–7.71(m,2H),7.48(dd,J＝8.8,2.7Hz,1H),7.03(d,J＝8.8Hz,1H).ESI-MS m/z 314.0(M-H) ^- .

Example 5: synthesis of N- (4-trifluoromethylphenyl) salicyloyl (IIIA-5)

IIIA-5 was prepared in the same manner as in example 1 except for using 2-hydroxybenzoic acid in place of 2-hydroxy-5-chloro-benzoic acid and 4-trifluoromethylaniline in place of 2-amino-4-trifluoromethylthiazole, with the following yields: 7.11 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ10.41(s,1H),9.48(s,1H),7.82(d,J＝8.4Hz,2H),7.66(d,J＝8.5Hz,2H),7.14(dd,J＝7.5,1.7Hz,1H),7.07(td,J＝7.7,1.8Hz,1H),6.80(dd,J＝8.1,1.2Hz,1H),6.75(td,J＝7.4,1.2Hz,1H),3.63(s,2H).ESI-MS m/z 280.0(M-H) ^- .

Example 6: synthesis of N- (4-trifluoromethoxyphenyl) -5-chlorosalicylamide (IIIA-6)

IIIA-6 was prepared in the same manner as in example 1 except that 4-trifluoromethoxyaniline was used in place of 2-amino-4-trifluoromethylthiazole and the reaction time was 5 hours, in such a yield that: 54.38 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.69(s,1H),10.53(s,1H),7.91(d,J＝2.7Hz,1H),7.85–7.77(m,2H),7.47(dd,J＝8.8,2.7Hz,1H),7.41–7.36(m,2H),7.02(d,J＝8.8Hz,1H).ESI-MS m/z 330.0(M-H) ^- .

Example 7: synthesis of N- (4-trifluoromethoxyphenyl) pyrazinamide (IIIA-7)

Compound IIIA-7 was prepared in the same manner as in example 1 except that pyrazine-2-carboxylic acid was used instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethoxyaniline was used instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 6 hours, in the following yield: 58.30 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ10.97(s,1H),9.31(d,J＝1.5Hz,1H),8.95(d,J＝2.5Hz,1H),8.83(dd,J＝2.5,1.5Hz,1H),8.06–7.99(m,2H),7.42–7.37(m,2H).ESI-MS m/z 284.0(M+H) ⁺ .

Example 8: synthesis of N- (4-trifluoromethylphenyl) pyrazinamide (IIIA-8)

Compound IIIA-8 was prepared in the same manner as in example 1 except for using pyrazine-2-carboxylic acid instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethylaniline instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 6 hours, with the yield of: 56.17 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.10(s,1H),9.32(d,J＝1.5Hz,1H),8.96(d,J＝2.5Hz,1H),8.84(dd,J＝2.5,1.5Hz,1H),8.15(d,J＝8.5Hz,2H),7.75(d,J＝8.6Hz,2H).ESI-MS m/z 269.0(M+H) ⁺ .

Example 9: synthesis of N- (3, 5-bis (trifluoromethyl) phenyl) -5-chlorosalicylamide (IIIA-9)

Compound IIIA-9 was prepared in the same manner as in example 1 except for using 3, 5-bis (trifluoromethyl) aniline instead of 2-amino-4-trifluoromethylthiazole and the reaction time was 7 hours, in the following yield: 44.38 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.38(s,1H),10.84(s,1H),8.84(m,2H),7.83-7.86(m,2H),7.47-7.51(m,1H),7.03-7.05(m,1H).ESI-MS m/z 382.0(M-H) ^- .

Example 10: synthesis of N- (4-trifluoromethylphenyl) -3, 5-dichlorosalicylamide (IIIA-10)

Compound IIIA-10 was prepared in the same manner as in example 1 except that 2-hydroxy-3, 5-dichlorobenzoic acid was used instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethylaniline was used instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 6 hours, in the following yield: 67.33 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ12.22(s,1H),10.88(d,J＝4.2Hz,1H),8.05(d,J＝2.5Hz,1H),7.93(d,J＝8.4Hz,2H),7.83(d,J＝2.5Hz,1H),7.77(d,J＝8.5Hz,2H).ESI-MS m/z 348.2.

Example 11: synthesis of N- (4-trifluoromethylphenyl) -2-cyano-5-chlorobenzamide (IIIA-11)

Compound IIIA-11 was prepared in the same manner as in example 1 except that 2-cyano-5-chlorobenzoic acid was used instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethylaniline was used instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 6 hours, in the following yield: 37.57 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ10.53(s,1H),8.34–8.28(m,1H),8.00–7.96(m,2H),7.90(d,J＝8.4Hz,2H),7.74(d,J＝8.2Hz,2H).ESI-MS m/z 323.1(M-H) ^- .

Example 12: synthesis of N- (4-trifluoromethoxyphenyl) -4-fluoro-5-chlorosalicylamide (IIIA-12)

Except that 2-hydroxy-5-chloro-benzoic acid is replaced by 2-hydroxy-4-fluoro-5-chlorobenzoic acid and 4-trifluoro-benzoic acidCompound IIIA-12 was prepared in the same manner as in example 1 except for substituting methoxyaniline for 2-amino-4-trifluoromethylthiazole and the reaction time was 10 hours, with the yield: 51.72 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ12.20(s,1H),10.51(s,1H),8.12(d,J＝8.5Hz,1H),7.86–7.79(m,2H),7.43–7.38(m,2H),7.04(d,J＝10.8Hz,1H).ESI-MS m/z 348.2(M-H) ^- .

Example 13: synthesis of N- (5-trifluoromethylthiazole) -4-fluoro-5-chlorosalicylamide (IIIA-13)

Compound IIIA-13 was prepared in the same manner as in example 1 except for using 2-hydroxy-4-fluoro-5-chlorobenzoic acid instead of 2-hydroxy-5-chloro-benzoic acid, 2-amino-5-trifluoromethylthiazole instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 3 hours, with the yield of: 51.47 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ12.65(s,2H),8.25(s,1H),8.05(d,J＝8.7Hz,1H),7.03(d,J＝8.7Hz,1H).ESI-MS m/z 339.1(M-H) ^- .

Example 14: synthesis of N- (4-trifluoromethylphenyl) -5-fluorosalicylamide (IIIA-14)

Compound IIIA-14 was prepared in the same manner as in example 1 except for using 2-hydroxy-5-fluorobenzoic acid instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethylaniline instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 9 hours, with the yield of: 53.29 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.55(s,1H),10.56(s,1H),7.86(d,J＝8.3Hz,2H),7.80(d,J＝2.7Hz,1H),7.77–7.73(m,2H),7.48(dd,J＝8.3,2.7Hz,1H),7.03(d,J＝8.8Hz,1H).ESI-MS m/z 298.0(M-H) ^- .

Example 15: synthesis of N- (4-trifluoromethylphenyl) -3, 5-difluorosalicylamide (IIIA-15)

Compound IIIA-15 was prepared in the same manner as in example 1 except that 2-hydroxy-3, 5-difluorobenzoic acid was used instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethylaniline was used instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 6 hours, in the following yield: 75.23 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.32(s,1H),10.72(s,1H),7.93(d,J＝8.5Hz,2H),7.76(d,J＝8.5Hz,2H),7.55(s,1H),7.53(d,J＝3.5Hz,1H).ESI-MS m/z 316.2(M-H) ^- .

Example 16: synthesis of N- (4-trifluoromethoxyphenyl) -3, 5-difluorosalicylamide (IIIA-16)

Compounds IIIA-16 were prepared in the same manner as in example 1 except that 2-hydroxy-3, 5-difluorobenzoic acid was used instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethoxyaniline was used instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 11 hours (whether or not other different reaction conditions exist), to obtain the following yields: 50.05 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.48(s,1H),10.60(s,1H),7.84–7.79(m,2H),7.60–7.52(m,2H),7.40(m,2H).ESI-MS m/z 332.2(M-H) ^- .

Example 17: synthesis of N- (3, 5-bis (trifluoromethyl) phenyl) -4-fluoro-5-chlorosalicylamide (IIIA-17)

Compound IIIA-17 was prepared in the same manner as in example 1 except that 2-hydroxy-4-fluoro-5-chlorobenzoic acid was used instead of 2-hydroxy-5-chloro-benzoic acid, 3, 5-bis (trifluoromethyl) aniline was used instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 9 hours, in the following yield: 65 percent. ¹ H NMR(500MHz,DMSO-d6)δ11.90(s,1H),10.85(s,1H),8.44(d,J＝1.6Hz,2H),8.07(d,J＝8.6Hz,1H),7.87(s,1H),7.05(d,J＝10.8Hz,1H).ESI-MS m/z 400.2(M-H) ^- .

Example 18: synthesis of N- (4-trifluoromethylphenyl) -4-fluoro-5-chlorosalicylamide (IIIA-18)

Compound IIIA-18 was prepared in the same manner as in example 1 except for using 2-hydroxy-4-fluoro-5-chlorobenzoic acid instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethylaniline instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 7 hours, with the yield of: 45.04 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ12.07(s,1H),10.63(s,1H),8.07(d,J＝8.6Hz,1H),7.93(d,J＝8.5Hz,2H),7.74(d,J＝8.5Hz,2H),7.02(d,J＝10.8Hz,1H).ESI-MS m/z 332.1(M-H) ^- .

Example 19: synthesis of N- (4-trifluoromethylphenyl) -2-hydroxy-5-chloronicotinamide (IIIA-19)

Compound IIIA-19 was prepared in the same manner as in example 1 except for using 2-hydroxy-5-chloronicotinic acid instead of 2-hydroxy-5-chloro-benzoic acid, 4-trifluoromethylaniline instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 12 hours, in the following yield: 69.72 percent. ¹ H NMR(500MHz,DMSO-d ₆ )δ13.25(s,1H),12.32(s,1H),8.36(d,J＝3.0Hz,1H),8.13(d,J＝3.1Hz,1H),7.90(d,J＝8.4Hz,2H),7.73(d,J＝8.5Hz,2H).ESI-MS m/z 315.0(M-H) ^- .

Example 20: synthesis of N- (2-chloro-4-trifluoromethylphenyl) -3,4, 5-trihydroxybenzamide (IIIA-20)

To an eggplant-shaped bottle were added 3,4, 5-trihydroxybenzoic acid (500 mg), methanol (10 mL) and sulfuric acid (100 uL), and the mixture was refluxed with stirring for 12 hours, after which the reaction of the starting materials was substantially completed. The reaction mixture was spin-dried, extracted and dried to give methyl 3,4, 5-trihydroxybenzoate (510 mg), which was directly used in the next step.

Acetone and potassium (2.8 g) were added to the above product, benzyl bromide (1.4 mL) was added dropwise, the mixture was refluxed at 66 ℃ for 6 hours, and the completion of the reaction of the starting materials was checked by TLC. Directly filtering the reaction solution, and spin-drying the filtrate to obtain 3,4, 5-tribenzyloxy methyl benzoate, and directly feeding into the next step.

To the above product was added methanol/water (3). And (3) post-treatment: removing methanol by rotary evaporation, extracting with ethyl acetate, adjusting pH to 5-6, separating, washing with water, washing with saturated saline solution, separating, drying, spin-drying the filtrate, and pulping with n-hexane to obtain light yellow 3,4, 5-tribenzyloxybenzoic acid (800 mg).

To a solanaceous bottle, a pale yellow solid (330 mg), dry toluene (8 mL), and 2-chloro-4-trifluoromethylaniline (78 uL) were sequentially added, and phosphorus trichloride (56 uL) was added at room temperature under nitrogen protection. At this time, the reaction mixture was milky white, and the mixture was refluxed at 115 ℃ overnight. To the reaction solution were added water, a small amount of sodium hydrogencarbonate solution, extracted with ethyl acetate, washed with saturated brine, and dried to elute with petroleum ether and ethyl acetate (100) to give 220mg of a white solid of N- (2-chloro-4-trifluoromethylphenyl) -3,4, 5-tribenzyloxybenzamide.

Dissolving white solid in methanol: tetrahydrofuran (1), palladium on carbon (20 mg) was added, hydrogen substitution was performed, the reaction was performed overnight, TLC detection was performed the next day, the reaction of the raw materials was complete, and the reaction was performed completely. And (3) post-treatment: celite assisted filtration, spin drying of the filtrate, sample mixing, elution with dichloromethane and methanol (50. ¹ H NMR(500MHz,DMSO-d ₆ )δ9.71(s,1H),9.24(s,2H),8.91(s,1H),8.00–7.93(m,2H),7.74(dd,J＝8.6,2.1Hz,1H),6.99(s,2H).ESI-MS m/z 453.1(M-H) ^- .

Example 21: synthesis of N- (4-trifluoromethoxyphenyl) -2,3, 4-trihydroxybenzamide (IIIA-21)

2,3, 4-Tribenzyloxybenzoic acid (330 mg), dry toluene (8 mL), and 4-trifluoromethoxyaniline (100 uL) were sequentially added to an eggplant-shaped bottle, and phosphorus trichloride (75 uL) was added thereto at room temperature under nitrogen protection. At the moment, the reaction solution is milky white suspension, the temperature is raised to 115 ℃, the reflux is carried out overnight, the clear TLC detection is carried out the next day, and the raw materials are completely reacted. To the reaction mixture was added water, a small amount of sodium hydrogencarbonate solution, extracted with ethyl acetate, washed with saturated brine, and dried, and eluted with petroleum ether and ethyl acetate (100.

Dissolving white solid in methanol: tetrahydrofuran (1), palladium on carbon (100 mg) was added, hydrogen substitution was performed, the reaction was performed overnight, TLC detection was performed the next day, the reaction of the raw materials was completed, and the reaction was completely performed. And (3) post-treatment: celite assisted filtration and spin dried, sample stirred and eluted with dichloromethane and methanol (50. ¹ H NMR(500MHz,DMSO-d ₆ )δ12.11(s,1H),10.27(s,1H),9.73(s,1H),8.57(s,1H),7.82–7.75(m,2H),7.43(d,J＝8.9Hz,1H),7.39–7.34(m,2H),6.42(d,J＝8.8Hz,1H).ESI-MS m/z 328.2(M-H) ^- .

Example 22: synthesis of N- (4-trifluoromethylphenyl) -2,3, 4-trihydroxybenzamide (IIIA-22)

2,3, 4-Tribenzyloxybenzoic acid (330 mg), dry toluene (8 mL), and 4-trifluoromethylaniline (94 uL) were sequentially added to an eggplant-shaped bottle, and phosphorus trichloride (78 uL) was added thereto at room temperature under nitrogen protection. At the moment, the reaction solution is milky white suspension, the temperature is raised to 115 ℃, the reflux is carried out overnight, the clear TLC detection is carried out the next day, and the raw materials are completely reacted. To the reaction mixture was added water, a small amount of sodium hydrogencarbonate solution, extracted with ethyl acetate, washed with saturated brine, and dried, and eluted with petroleum ether and ethyl acetate (100.

Dissolving the white solid in methanol: tetrahydrofuran (1), palladium on carbon (100 mg) was added, hydrogen substitution was performed, the reaction was performed overnight, TLC detection was performed the next day, the reaction of the raw materials was completed, and the reaction was completely performed. And (3) post-treatment: celite assisted filtration and spin dried, sample stirred and eluted with dichloromethane and methanol (50. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.91(s,1H),10.40(s,1H),9.79(s,1H),8.62(s,1H),7.92(d,J＝8.5Hz,2H),7.72(d,J＝8.5Hz,2H),7.45(d,J＝8.8Hz,1H),6.43(d,J＝8.8Hz,1H).ESI-MS m/z 312.2(M-H) ^- .

Example 23: synthesis of N- (4-trifluoromethylphenyl) -2-methoxy-5-chlorobenzamide (IIIA-23)

To an eggplant-shaped bottle were added 5-chloro-2-methoxybenzoic acid (186 mg), dry toluene (5 ml), and p-trifluoromethylaniline (125 uL) in this order, and phosphorus trichloride (105 uL) was added at room temperature under nitrogen protection. At this time, the reaction solution was a white suspension, heated to 115 ℃ and refluxed overnight, and the completion of the reaction of the starting materials was checked by TLC. To the reaction mixture was added water, a small amount of sodium hydrogencarbonate solution, extracted with ethyl acetate, washed with saturated brine, dried, and eluted with petroleum ether and ethyl acetate (100. ¹ H NMR(500MHz,DMSO-d6)δ10.54(s,1H),7.93(d,J＝8.4Hz,2H),7.71(d,J＝8.5Hz,2H),7.61(d,J＝2.7Hz,1H),7.56(dd,J＝8.9,2.8Hz,1H),7.22(d,J＝8.9Hz,1H),3.88(s,3H).ESI-MS m/z 328.0(M-H) ^- .

Example 24: synthesis of N- (4-trifluoromethylphenyl) -5-chloro-2-acetamidobenzamide (IIIA-24)

2-amino-5-chlorobenzoic acid (1.368 g) was dissolved in acetic acid (15 mL), acetic anhydride (1.34 mL) was added, the reaction was complete after 30 minutes by TLC at room temperature to 75 ℃. And (3) post-treatment: pouring the reaction solution into ice water, stirring, carrying out suction filtration, washing a filter cake with water, and drying to obtain the 2-acetamido-5-chlorobenzoic acid until the next step.

To the solid was added dry toluene, p-trifluoromethylaniline (878 uL), phosphorus trichloride (800 uL), and refluxing overnight at 115 ℃ until the next day, which was not dissolved to be about 90% reaction. And (3) post-treatment: water and a small amount of sodium bicarbonate solution were added to the reaction mixture, toluene was removed by rotary evaporation, extraction was performed with ethyl acetate, and the mixture was washed with saturated brine, dried and eluted with petroleum ether and ethyl acetate (100. ¹ H NMR(500MHz,DMSO-d ₆ )δ8.06(d,J＝2.5Hz,1H),7.99(d,J＝8.2Hz,2H),7.91(d,J＝8.7,1H),7.77(d,J＝8.2Hz,2H),7.73(d,J＝8.7Hz,1H),2.14(s,3H).ESI-MS m/z 355.1(M-H) ^- .

Example 25: synthesis of N- (4-trifluoromethoxyphenyl) -5-chloro-2-acetamidobenzamide (IIIA-25)

A round-bottomed flask was charged with the white solid (277 mg) obtained in the first step in example 24, dried toluene (5 mL), p-trifluoromethoxyaniline (171.6 uL), phosphorus trichloride (146 uL), nitrogen substitution, and stirring at room temperature to 115 ℃ for 12 hours, so that the reaction phase was always cloudy, and after cooling, the starting material was reacted by about 90% by TLC. And (3) post-treatment: adding a small amount of methanol to quench the reaction, removing partial toluene by rotary evaporation, adding ethyl acetate and water into the system, layering, separating, washing an organic phase by saturated saline solution, drying, filtering, mixing a sample, eluting by petroleum ether and ethyl acetate (100): methanol =30:1, obtaining the target compound IIIA-25, 304mg with correct nuclear magnetism. ¹ H NMR(500MHz,DMSO-d ₆ )δ8.04(s,1H),7.90(d,J＝8.7Hz,1H),7.73(d,J＝8.6Hz,1H),7.65(d,J＝8.3Hz,2H),7.60(d,J＝8.3Hz,2H),2.15(s,3H).ESI-MS m/z 395.0(M+Na) ⁺ .

Example 26: n-, (Synthesis of 4-trifluoromethylphenyl) -5-chloro-3-acetylsalicylamide (IIIA-26)

5-Chlorosalicylic acid (5 g), dichloromethane (15 mL), and DMF (10 drops) were added to the flask in this order, oxalyl chloride (3.5 mL) was added dropwise to the ice bath, and the reaction was allowed to proceed to room temperature for about 3 hours to completion. The solution was added dropwise to methanol (25 mL), and after completion of the addition, the mixture was stirred for 10 minutes and then concentrated, ethyl acetate and water were used for liquid separation, and the organic phase was washed with saturated brine, dried, filtered, and the filtrate was concentrated to give methyl 5-chloro-2-hydroxybenzoate (5.1 g) for addition.

To methyl 5-chloro-2-hydroxybenzoate (5.1 g) obtained in the previous step, acetic anhydride (5 mL), sulfuric acid (100 uL) were added, and the mixture was stirred at room temperature for 30 minutes. Ethyl acetate (15 mL) and saturated sodium bicarbonate solution (added slowly in portions) were added to the reaction until no air bubbles were formed after the addition. Separating, washing the organic phase with saturated salt solution, drying, filtering, concentrating to obtain 5-chloro-2-acetoxyl methyl benzoate, and putting into the next step.

Transferring the 5-chloro-2-acetoxyl methyl benzoate into a double-mouth bottle, adding aluminum trichloride (30 g) and sodium chloride (16 g), mechanically stirring for 3 hours at 120 ℃ under the protection of nitrogen, and detecting by TLC that the raw materials are basically reacted completely. Adding 1M hydrogen chloride solution (slowly adding) into the reaction system under ice bath, adding ethyl acetate (30 mL) after stirring uniformly, extracting and separating liquid, and extracting the water layer for multiple times. The combined organic phases were washed with brine, dried, filtered, concentrated and stirred, and eluted with petroleum ether and ethyl acetate (10.

5-chloro-3-acetyl 2-hydroxybenzoic acid (990 mg), dry toluene (25 mL), p-trifluoromethylaniline (554 uL), phosphorus trichloride (446 uL), and nitrogen were added to the reaction flask in this order, and the temperature was raised from room temperature to 115 ℃ for 10 hours. TLC showed the reaction was not complete. And (3) post-treatment: a small amount of methanol was added to the reaction system, concentrated, and the sample was stirred and eluted with petroleum ether and ethyl acetate (100. ¹ H NMR(500MHz,DMSO-d ₆ )δ13.01(s,1H),10.80(s,1H),8.13(d,J＝2.7Hz,1H),8.06(d,J＝2.8Hz,1H),7.95(d,J＝8.3Hz,2H),7.76(d,J＝8.4Hz,2H),2.74(s,3H).ESI-MS m/z 356.1(M-H) ^- .

Example 27: synthesis of N- (4-trifluoromethoxyphenyl) -5-chloro-3-acetylsalicylamide (IIIA-27)

The solid obtained in the third step of example 26 (85 mg), dry toluene (2 mL), p-trifluoromethoxyaniline (54 uL), phosphorus trichloride (45 uL), and nitrogen were added to a reaction flask in this order, and the temperature was raised from room temperature to 115 ℃ for reaction for 13 hours. TLC showed the reaction was not complete. And (3) post-treatment: a small amount of methanol was added to the reaction system, concentrated, extracted with ethyl acetate and water, washed with saturated brine, dried, and eluted with petroleum ether and ethyl acetate (100. ¹ H NMR(500MHz,DMSO-d ₆ )δ13.07(s,1H),10.72(s,1H),8.09–8.04(m,2H),7.85–7.79(m,2H),7.39(d,J＝8.6Hz,2H),2.71(s,3H).ESI-MS m/z 372.0(M-H) ^- .

Example 28: synthesis of N- (4-trifluoromethylphenyl) -5-chloro-3-nitrosalicylamide (IIIA-28)

5-chloro-salicylic acid (342 mg) is dissolved in concentrated sulfuric acid, ice bath is carried out, concentrated nitric acid (232 mg) is added, after ice bath reaction for 4 hours, TLC detection is carried out, and the raw materials are completely reacted. Dropping the reaction solution into ice water, stirring for 30 minutes, filtering, washing a filter cake with water, drying and concentrating to obtain the 3-nitro-5-chloro-2-hydroxybenzoic acid, and putting the mixture into the next step.

3-Nitro-5-chloro-2-hydroxybenzoic acid (310 mg), p-trifluoromethylaniline (230 mg), dry toluene (20 mL), phosphorus trichloride (149 uL), at 115 ℃ under reflux. Adding water and a small amount of sodium bicarbonate solution into the reaction solution, removing toluene by rotary evaporation, extracting with ethyl acetate, washing with saturated saline, drying, and adding petroleum ether and ethyl acetateEthyl acid (100. ¹ H NMR(500MHz,DMSO-d ₆ )δ11.83(s,1H),8.17–8.15(m,1H),8.14–8.10(m,1H),7.91(d,J＝8.5Hz,2H),7.75(d,J＝8.4Hz,2H).ESI-MS m/z 359.0(M-H) ^- .

Example 29: synthesis of N- (4-trifluoromethylmercaptophenyl) -4-fluoro-5-chlorosalicylamide (IIIA-29)

Compound IIIA-29 was prepared in the same manner as in example 1 except that 2-hydroxy-4-fluoro-5-chlorobenzoic acid was used instead of 2-hydroxy-5-chloro-benzoic acid and 4-trifluoromethylmercaptoaniline was used instead of 2-amino-4-trifluoromethylthiazole for 5 hours, to give compound IIIA-29 in the following yield: 69.23 percent. ¹ H NMR(400MHz,DMSO-d6)δ12.08(s,1H),10.58(s,1H),8.06(d,J＝8.6Hz,1H),7.90–7.83(m,2H),7.76–7.69(m,2H),7.02(d,J＝10.8Hz,1H).ESI-MS m/z363.0(M-H) ^- .

Example 30: synthesis of N- (3-trifluoromethyl-4-cyanophenyl) -4-fluoro-5-chlorosalicylamide (IIIA-30)

Compound IIIA-30 was prepared in the same manner as in example 1 except for using 2-hydroxy-4-fluoro-5-chlorobenzoic acid instead of 2-hydroxy-5-chloro-benzoic acid and 3-trifluoromethyl-4-cyanoaniline instead of 2-amino-4-trifluoromethylthiazole, with the following yield: 37.23 percent. ¹ H NMR(500MHz,DMSO-d6)δ11.83(s,1H),10.90(s,1H),8.40(s,1H),8.16(s,2H),7.99(d,J＝8.6Hz,1H),7.02(d,J＝10.8Hz,1H).ESI-MS m/z 358.0(M-H) ^- .

Example 31: synthesis of N- (3-trifluoromethyl-4-cyanopyridine) -4-fluoro-5-chlorosalicylamide (IIIA-31)

Compound IIIA-31 was prepared in the same manner as in example 1 except for using 2-hydroxy-4-fluoro-5-chlorobenzoic acid instead of 2-hydroxy-5-chloro-benzoic acid and 2-cyano-3-trifluoromethyl-5-aminopyridine instead of 2-amino-4-trifluoromethylthiazole and the reaction time was 6 hours, in the following yield: 36.13 percent. ¹ H NMR(500MHz,DMSO-d6)δ11.90(s,1H),11.16(s,1H),9.24(d,J＝2.3Hz,1H),8.82(d,J＝2.3Hz,1H),7.99(d,J＝8.6Hz,1H),7.03(d,J＝10.8Hz,1H).ESI-MS m/z 359.0(M-H) ^- .

Example 32: synthesis of N- (3-chloro-4-trifluoromethyl) -5-chlorosalicylamide (IIIA-32)

Compound IIIA-32 was prepared in the same manner as in example 1 except that 2-hydroxy-5-chlorobenzoic acid was used instead of 2-hydroxy-5-chloro-benzoic acid, 3-chloro-4-trifluoromethylaniline was used instead of 2-amino-4-trifluoromethylthiazole, and the reaction time was 5 hours, in the following yield: 40.23 percent. ¹ H NMR(500MHz,DMSO-d6)δ11.92(s,1H),10.78(d,J＝6.0Hz,1H),8.14(d,J＝1.9Hz,1H),8.02(d,J＝8.6Hz,1H),7.89–7.81(m,2H),7.01(d,J＝10.8Hz,1H).ESI-MS m/z 348.01(M-H) ^- .

Example 33: synthesis of (4-chloro-5-fluoro-2- (((4- (trifluoromethyl) phenyl) carbamoyl) phenoxy) methyl isopropyl carbonate (IIIB-1)

Acetone (1 mL), chloromethyl isopropyl carbonate (46 mg), and sodium iodide (45 mg) were added to the reaction flask in this order, and stirred at 60 ℃ for 3 hours. Filtered and the filter cake washed with a small amount of acetone.

Potassium carbonate (40 mg), compound IIIA-18 (70 mg), acetone (2 mL), and DMF (10 drops) were sequentially added to the reaction flask to aid dissolution, the filtrate obtained in the above step was added with stirring, and the reaction was checked by TLC for completion by stirring for 2 hours. Concentrating the reaction solution, adding acetic acid to the concentrateEthyl ester, water, adjusting pH of water phase with saturated ammonium chloride solution, separating, washing organic phase with saturated saline solution, drying, filtering, concentrating, pulping concentrate with petroleum ether (1.5 mL) and ethyl acetate (10 drops), and filtering to obtain compound IIIB-1 60mg with correct nuclear magnetism. ¹ H NMR(500MHz,DMSO-d ₆ )δ10.60(s,1H),7.90(d,J＝8.4Hz,2H),7.88(d,J＝8.4Hz,1H),7.74(d,J＝8.6Hz,2H),7.55(d,J＝8.6Hz,1H),5.88(s,2H),4.76(m,J＝6.3Hz,1H),1.20(d,J＝6.3Hz,6H).ESI-MS m/z 447.8(M-H) ^- .

Example 34 Synthesis of 5-chloro-4-fluoro-2- ((5-methyl-2-oxo-1, 3-dioxolan-4-yl) methoxy) -N- (4- (trifluoromethyl) phenyl) benzamide (IIIB-2)

Acetone (1 mL), 4-chloromethyl-5-methyl-1, 3-dioxol-2-one (44 m), and sodium iodide (45 mg) were added to the reaction flask in this order, and stirred at 60 ℃ for 3 hours. Filtration and washing of the filter cake with a small amount of acetone.

Sequentially adding potassium carbonate (50 mg), compound IIIA-18 (70 mg), acetone (2 mL) and DMF (10 drops) into a reaction bottle for dissolving assistance, adding the filtrate obtained in the step a under stirring, and stirring for 5 hours to detect the residual reaction raw materials by TLC. And (3) post-treatment: spin-drying the organic solvent, adding ethyl acetate and water to the concentrate, adjusting the pH of the aqueous phase to acidic with saturated ammonium chloride, separating, washing the organic phase with saturated saline solution, drying, filtering, concentrating, pulping the concentrate with petroleum ether (1.5 mL) and ethyl acetate (10 drops), filtering to obtain compound IIIB-2 56mg, correct nuclear magnetism. ¹ H NMR(500MHz,DMSO-d ₆ )δ10.53(s,1H),7.89(d,J＝8.4Hz,2H),7.84(d,J＝8.5Hz,1H),7.72(d,J＝8.6Hz,2H),7.57(d,J＝11.4Hz,1H),5.18(s,2H),2.18(s,3H).ESI-MS m/z 444.1(M-H) ^- .

Example 35: synthesis of N- (4-trifluoromethylphenyl) -2-acetoxy-4-fluoro-5-chlorobenzamide (IIIB-3)

The weighed salicylamide derivative IIIA-18 was added to the eggplant-shaped bottle in this order, acetic anhydride (0.5 mL) and concentrated sulfuric acid (1 drop) were added thereto, and the mixture was stirred at room temperature for 45 minutes, and TLC detection was carried out to detect almost complete conversion of the starting material. To the reaction solution were added ethyl acetate (4 mL), ice water (4 mL), extracted and separated, and the ethyl acetate layer was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and stirred to elute with petroleum ether and ethyl acetate (100) to obtain the corresponding target compound IIIB-3 in the yield: 30 percent. ¹ H NMR(500MHz,DMSO-d6)δ10.79(s,1H),8.04(d,J＝8.1Hz,1H),7.89(d,J＝8.4Hz,2H),7.73(d,J＝8.5Hz,3H),7.57(d,J＝9.7Hz,1H),2.21(s,3H).

Example 36: synthesis of N- (4-trifluoromethylphenyl) -2-acetoxy-3, 5-difluorobenzamide (IIIB-4)

IIIB-4 was prepared in the same manner as in example 35 except that the compound IIIA-15 was used in place of IIIA-18 in the following yield: and 72 percent. ¹ H NMR(500MHz,DMSO-d6)δ10.89(s,1H),7.91(d,J＝8.4Hz,2H),7.79–7.69(m,3H),7.57(ddd,J＝8.3,2.9,1.6Hz,1H),2.28(s,3H).

Example 37: synthesis of N- (4-trifluoromethoxyphenyl) -2-acetoxy-4-fluoro-5-chlorobenzamide (IIIB-5)

IIIB-5 was prepared in the same manner as in example 35 except that compound IIIA-12 was used instead of IIIA-18, in the yield: 34 percent. ¹ H NMR(500MHz,DMSO-d6)δ10.64(s,1H),8.01(d,J＝8.2Hz,1H),7.81–7.75(m,2H),7.56(d,J＝9.8Hz,1H),7.42–7.31(m,2H),2.21(s,3H).

Example 38: synthesis of N- (4-trifluoromethylphenyl) -2-isobutyroyloxy-4-fluoro-5-chlorobenzamide (IIIB-6)

IIIB-6 was prepared in the same manner as in example 35 except that isobutyric anhydride was used in place of acetic anhydride and the reaction time was 1 hour, in the yield: 74 percent. ¹ H NMR(500MHz,DMSO-d6)δ10.82(s,1H),8.02(d,J＝8.1Hz,1H),7.90(d,J＝8.4Hz,2H),7.74(d,J＝8.6Hz,2H),7.60(d,J＝9.8Hz,1H),2.76(m,1H),1.15(d,J＝7.0Hz,5H).

Example 39: synthesis of N- (4-trifluoromethylphenyl) -2-isobutyryloxy-3, 5-difluorobenzamide (IIIB-7)

IIIB-7 was prepared in the same manner as in example 35 except that the compound IIIA-15 was used in place of IIIA-18, isobutyric anhydride was used in place of acetic anhydride, and the reaction time was 1 hour, in the yield: 76 percent. ¹ H NMR(500MHz,DMSO-d6)δ10.88(s,1H),7.89(d,J＝8.4Hz,2H),7.74(d,J＝8.8Hz,2H),7.72–7.69(m,1H),7.56–7.51(m,1H),2.82(m,1H),1.15(d,J＝7.0Hz,5H).

Example 40: synthesis of N- (4-trifluoromethoxyphenyl) -2-isobutyryloxy-4-fluoro-5-chlorobenzamide (IIIB-8)

IIIB-8 was prepared in the same manner as in example 36 except that compound IIIA-12 was used in place of IIIA-18, isobutyric anhydride was used in place of acetic anhydride, and the reaction time was 1 hour, in the yield: 65 percent. ¹ H NMR(500MHz,DMSO-d6)δ10.64(s,1H),7.98(d,J＝8.2Hz,1H),7.80–7.75(m,2H),7.58(d,J＝9.8Hz,1H),7.37(d,J＝8.6Hz,2H),2.75(m,1H),1.14(d,J＝7.0Hz,5H).

Example 41: synthesis of N- (4-trifluoromethylphenyl) -2- (L-alanine) oxy-4-fluoro-5-chlorobenzamide hydrochloride (IIIB-9)

IIIA-18 (150 mg), N-triphenylmethyl-L-alanine (150 mg), and N, N-dimethylformamide (3 mL) were sequentially added to an eggplant-shaped bottle, and the mixture was stirred at-10 ℃ and added with cyclohexyl carbodiimide (94 mg), and the mixture was reacted at this temperature for 1 hour, and then, the reaction was switched to room temperature for 6 hours. During the process, white solid is separated out, filtered, the filtrate is extracted by ethyl acetate and water, an ethyl acetate layer is washed by saturated sodium chloride solution, dried and eluted by petroleum ether and ethyl acetate (50).

IIIB-9-1 obtained in the previous step was dissolved in methylene chloride (1.5 mL), and trifluoroacetic acid (0.3 mL) was added thereto in an ice bath and stirred for 1 hour. Spin-drying the solvent, extracting with ethyl acetate and water, washing with saturated sodium chloride solution, drying, concentrating, and eluting with petroleum ether and ethyl acetate (2). Methanol was added to the free compound, 1.5 equivalents of methanolic hydrogen chloride was added, the mixture was stirred for 15 minutes and concentrated, and then water was added to toluene (3 times) to obtain IIIB-9 70mg. ¹ H NMR(500MHz,DMSO-d6)δ12.73(s,1H),10.54(s,1H),9.09(d,J＝6.6Hz,1H),8.26(d,J＝8.6Hz,1H),7.82(d,J＝8.5Hz,2H),7.69(d,J＝8.6Hz,2H),7.01(d,J＝10.8Hz,1H),4.65(p,J＝7.0Hz,1H),1.47(d,J＝7.1Hz,3H).ESI-MS m/z 403.04(M-H) ^- .

Example 42: synthesis of N- (4-trifluoromethylphenyl) -2- (L-alanine) oxy-3, 5-difluorobenzamide hydrochloride (IIIB-10)

IIIA-15 (143 mg), N-triphenylmethyl-L-alanine (150 mg), and N, N-dimethylformamide (3 mL) were sequentially added to an eggplant-shaped bottle, and the mixture was stirred at-10 ℃ and added with cyclohexyl carbodiimide (94 mg), and the reaction was carried out for 1 hour while maintaining the temperature, and then the reaction was carried out for 6 hours while changing to room temperature. During this time, a white solid precipitated, filtered, the filtrate was extracted with ethyl acetate and water, the ethyl acetate layer was washed with saturated sodium chloride solution, dried, and eluted with petroleum ether and ethyl acetate (50.

IIIB-10-1 obtained in the previous step was dissolved in methylene chloride (1.5 mL), and trifluoroacetic acid (0.3 mL) was added thereto in an ice bath and stirred for 1 hour. Spin-drying the solvent, extracting with ethyl acetate and water, washing with saturated sodium chloride solution, drying, concentrating, and eluting with petroleum ether and ethyl acetate (2. Methanol was added to the free compound, 1.5 equivalents of methanolic hydrogen chloride was added, the mixture was stirred for 15 minutes and concentrated, and then water was added to toluene (3 times) to obtain IIIB-10 70mg. ¹ H NMR(500MHz,DMSO-d6)δ12.26(s,1H),10.55(s,1H),9.19(d,J＝6.6Hz,1H),7.83(d,J＝8.6Hz,2H),7.74(dt,J＝10.0,2.4Hz,1H),7.69(d,J＝8.6Hz,2H),7.55(ddd,J＝11.3,8.4,3.1Hz,1H),4.67(p,J＝7.0Hz,1H),1.48(d,J＝7.1Hz,3H).ESI-MS m/z 387.04(M-H) ^- .

Example 43: synthesis of N- (4-trifluoromethoxyphenyl) -2- (L-alanine) oxy-4-fluoro-5-chlorobenzamide hydrochloride (IIIB-11)

IIIA-12 (136 mg), N-triphenylmethyl-L-alanine (132 mg), and N, N-dimethylformamide (3 mL) were sequentially added to an eggplant-shaped bottle, and the mixture was stirred at-10 ℃ and added with cyclohexylcarbodiimide (83 mg), and the mixture was reacted at this temperature for 1 hour, and then, the reaction was switched to room temperature for 6 hours. During the process, white solid is separated out, filtered, the filtrate is extracted by ethyl acetate and water, an ethyl acetate layer is washed by saturated sodium chloride solution, dried and eluted by petroleum ether and ethyl acetate (50.

IIIB-11-1 obtained in the previous step was dissolved in methylene chloride (1.5 mL), and trifluoroacetic acid (0.3 mL) was added thereto in an ice bath and stirred for 1 hour. The solvent was dried, extracted with ethyl acetate, water, washed with saturated sodium chloride solution, dried, concentrated and eluted with petroleum ether and ethyl acetate (2). Methanol was added to the free compound, 1.5 equivalents of methanolic hydrogen chloride was added thereto, and the mixture was stirred for 15 minutesConcentration and addition of toluene with water (3 times) gave IIIB-11 mg of 70mg. ¹ H NMR(500MHz,DMSO-d6)δ12.73(s,1H),10.36(s,1H),9.08(d,J＝6.7Hz,1H),8.25(d,J＝8.6Hz,1H),7.74–7.70(m,2H),7.33(d,J＝8.7Hz,2H),7.00(d,J＝10.8Hz,1H),4.63(p,J＝7.0Hz,1H),1.45(d,J＝7.0Hz,3H).ESI-MS m/z 419.02(M-H) ^- .

Example 44: synthesis of N- (4-trifluoromethylphenyl) -2- (L-alanine) oxy-4-fluoro-5-chlorobenzamide hydrochloride (IIIB-12)

IIIA-18 (166 mg), N-t-butyloxycarbonyl-L-valine (108 mg) and N, N-dimethylformamide (3 mL) were sequentially added to an eggplant-shaped flask, and stirred at-10 ℃ to add cyclohexylcarbodiimide (155 mg), and the reaction was carried out for 1 hour while maintaining the temperature, and then the reaction was carried out for 6 hours while changing to room temperature. During this time, a white solid precipitated, filtered, the filtrate was extracted with ethyl acetate, water, the ethyl acetate layer was washed with saturated sodium chloride solution, dried, and eluted with petroleum ether and ethyl acetate (50.

IIIB-12-1 obtained in the previous step was dissolved in methylene chloride (1.5 mL), and trifluoroacetic acid (0.3 mL) was added thereto in an ice bath and stirred for 1 hour. Spin-drying the solvent, extracting with ethyl acetate and water, washing with saturated sodium chloride solution, drying, concentrating, and eluting with petroleum ether and ethyl acetate (2. Methanol was added to the free compound, 1.5 equivalents of methanolic hydrogen chloride was added, the mixture was stirred for 15 minutes and concentrated, and then water was added to toluene (3 times) to obtain IIIB-12 mg. ¹ H NMR(500MHz,DMSO-d6)δ12.60(s,1H),10.65(s,1H),8.93(d,J＝8.1Hz,1H),8.23(d,J＝8.7Hz,1H),7.85(d,J＝8.5Hz,2H),7.78–7.68(m,2H),7.00(d,J＝10.7Hz,1H),4.57(t,J＝7.6Hz,1H),2.21(h,J＝6.8Hz,1H),0.99(dd,J＝6.7,1.5Hz,6H).ESI-MS m/z431.05(M-H) ^- .

Example 45: synthesis of N- (4-trifluoromethoxyphenyl) -2- (L-valine) oxy-4-fluoro-5-chlorobenzamide hydrochloride (IIIB-13)

IIIA-12 (170 mg), N-t-butoxycarbonyl-L-valine (105 mg), and N, N-dimethylformamide (3 mL) were sequentially added to an eggplant-shaped flask, and stirred at-10 ℃ to add cyclohexylcarbodiimide (83 mg), the temperature was maintained, the reaction was carried out for 1 hour, and then the reaction was carried out at room temperature for 6 hours. During the process, white solid is separated out, filtered, the filtrate is extracted by ethyl acetate and water, an ethyl acetate layer is washed by saturated sodium chloride solution, dried and eluted by petroleum ether and ethyl acetate (50.

IIIB-13-1 obtained in the previous step was dissolved in methylene chloride (1.5 mL), and trifluoroacetic acid (0.3 mL) was added thereto in an ice bath and stirred for 1 hour. The solvent was dried, extracted with ethyl acetate, water, washed with saturated sodium chloride solution, dried, concentrated and eluted with petroleum ether and ethyl acetate (2). To the free compound, methanol was added, 1.5 equivalents of methanolic hydrogen chloride was added, the mixture was stirred for 15 minutes, concentrated, and then charged with toluene and water (3 times) to obtain IIIB-13 mg. ¹ H NMR(500MHz,DMSO-d6)δ12.63–12.60(m,1H),10.48(s,1H),8.93(d,J＝8.2Hz,1H),8.23(d,J＝8.7Hz,1H),7.76–7.71(m,2H),7.35(d,J＝8.6Hz,2H),6.99(d,J＝10.7Hz,1H),4.54(t,J＝7.7Hz,1H),2.20(h,J＝6.6Hz,1H),0.98(dd,J＝6.8,1.6Hz,6H).ESI-MS m/z 447.08(M-H) ^- 。

Example 46: synthesis of (4-chloro-5-fluoro-2- (((4- (trifluoromethoxy) phenyl) carbamoyl) phenoxy) methyl isopropyl carbonate (IIIB-14)

Acetone (1 mL), chloromethyl isopropyl carbonate (61 mg), and sodium iodide (62 mg) were added to the reaction flask in this order, and the mixture was stirred at 60 ℃ for 3 hours. Filtration and washing of the filter cake with a small amount of acetone.

Potassium carbonate (66 mg), compound IIIA-12 (70 mg), acetone (2 mL), DMF (10 drops) were added to the reaction flask in sequence to aid dissolution, and the mixture was stirred and addedThe filtrate was stirred for 2 hours to check the completion of the reaction by TLC. Concentrating the reaction solution, adding ethyl acetate and water into the concentrate, adjusting the pH of the water phase by using a saturated ammonium chloride solution, separating liquid, washing an organic phase by using saturated saline solution, drying, filtering, concentrating, pulping the concentrate by using petroleum ether (1.5 mL) and ethyl acetate (10 drops), filtering to obtain a compound IIIB-14 70mg, and performing nuclear magnetic correction. ¹ H NMR(500MHz,DMSO-d6)δ10.41(s,1H),7.85(d,J＝8.4Hz,1H),7.83–7.77(m,2H),7.53(d,J＝11.0Hz,1H),7.38(d,J＝8.7Hz,2H),5.88(s,2H),4.76(hept,J＝6.2Hz,1H),1.20(d,J＝6.2Hz,6H).

Example 47: synthesis of N- (4-trifluoromethoxyphenyl) -2- (valinemenoxy) -4-fluoro-5-chlorobenzamide hydrochloride (IIIB-15)

Acetone (3 mL), (S) -2- ((tert-butoxycarbonyl) amino) -3-methylbutyrate chloromethyl ester (397 mg), sodium iodide (220 mg) were added in this order to the reaction flask, and stirred at 60 ℃ for 3 hours. Filtration and washing of the filter cake with a small amount of acetone.

To a reaction flask were added potassium carbonate (66 mg), compound IIIA-12 (349 mg), and acetone (10 mL) in this order, and the filtrate obtained in the above step was added with stirring and stirred for 12 hours. The reaction mixture was concentrated, ethyl acetate and water were added to the concentrate, the pH of the aqueous phase was adjusted with a saturated ammonium chloride solution, and the organic phase was washed with a saturated saline solution, dried, filtered, concentrated, and sample-mixed, and eluted with petroleum ether and ethyl acetate (2).

IIIB-15-1 was dissolved in dichloromethane (6 mL), and trifluoroacetic acid (0.75 mL) was added under ice bath, and after addition, the mixture was stirred at room temperature for 3 hours. Concentrating, extracting with dichloromethane and water, adjusting pH of water layer to alkaline, washing with saturated sodium chloride solution, filtering, and concentrating. Methanol was added to the concentrate, 1.5 equivalents of methanolic hydrogen chloride was added, the mixture was stirred for 15 minutes and concentrated, and then water was added to toluene (3 times) to obtain IIIB-15 mg. The nuclear magnetism is correct. ¹ H NMR(500MHz,)δ10.49(d,J＝3.6Hz,1H),8.49(s,2H),7.88–7.79(m,3H),7.56(d,J＝11.0Hz,1H),7.37(d,J＝8.6Hz,2H),6.06(d,J＝6.9Hz,1H),6.01(d,J＝6.8Hz,1H),4.01(s,1H),2.13(dq,J＝11.4,6.9Hz,1H),0.89(t,J＝7.2Hz,6H).ESI-MS m/z 479.1(M+H) ^- 。

Example 48: synthesis of N- (4-trifluoromethoxyphenyl) -2- (4-methylpiperazinoyloxy) -4-fluoro-5-chlorobenzamide hydrochloride (IIIB-16)

Compound IIIA-12 (175 mg), 4-dimethylaminopyridine (18 mg), 4-methylpiperazine-1-carbonyl chloride hydrochloride (199 mg) and pyridine were sequentially added to a reaction flask and refluxed for 3 hours. After concentration, the mixture was extracted with ethyl acetate and water, and the aqueous layer was washed with saturated ammonium chloride, dried over anhydrous sodium sulfate, and then stirred and eluted with petroleum ether and ethyl acetate (1. ¹ H NMR(500MHz,DMSO-d6)δ10.59(s,1H),7.91(d,J＝8.2Hz,1H),7.80–7.74(m,2H),7.56(d,J＝10.0Hz,1H),7.38(d,J＝8.6Hz,2H),3.50(s,2H),3.31(s,2H),2.14(s,4H),2.05(s,3H).ESI-MS m/z 476.1(M+H) ^- 。

Example 49: synthesis of (4, 6-difluoro-2- (((4- (trifluoromethoxy) phenyl) carbamoyl) phenoxy) methyl isopropyl carbonate (IIIB-17)

Acetone (1 mL), chloromethyl isopropyl carbonate (61 mg), and sodium iodide (62 mg) were added to the reaction flask in this order, and the mixture was stirred at 60 ℃ for 3 hours. Filtered and the filter cake washed with a small amount of acetone.

Potassium carbonate (66 mg), compound IIIA-15 (85 mg), acetone (2 mL), and DMF (10 drops) were sequentially added to the reaction flask to aid dissolution, the filtrate obtained in the above step was added with stirring, and the reaction was checked by TLC for completion by stirring for 2 hours. Concentrating the reaction solution, adding ethyl acetate and water into the concentrate, adjusting pH of the water phase with saturated ammonium chloride solution, separating, washing the organic phase with saturated saline solution, drying, filtering, concentrating, pulping the concentrate with petroleum ether (1.5 mL) and ethyl acetate (10 drops), and filteringThe compound IIIB-17 mg is obtained by filtration. ¹ H NMR(500MHz,DMSO-d6)δ10.74(s,1H),7.90(d,J＝8.4Hz,2H),7.74(d,J＝8.5Hz,2H),7.64(ddd,J＝11.3,8.5,3.1Hz,1H),7.40(dt,J＝8.2,2.3Hz,1H),5.62(s,2H),4.61(hept,J＝6.3Hz,1H),1.12(d,J＝6.2Hz,6H).

Example 50:3- ((((4-chloro-5-fluoro-2- (4- (trifluoromethoxy) phenylcarbamoyl) phenoxy) methoxy) carbonyloxy) -N-methylpropane-1-ammonium chloride)

Acetone (3 mL), 3- (N-methyl-N-t-butoxycarbonyl) propyl chloromethyl carbonate (422mg, 1.5 mmol), sodium iodide (300mg, 2mmol) were added to the reaction flask in this order, and the mixture was stirred at 60 ℃ for 3 hours. Filtered and the filter cake washed with a small amount of acetone.

To a reaction flask were added potassium carbonate (276 mg), compound IIIA-12 (349 mg), acetone (10 mL), and DMF (10 drops) in this order, and the filtrate obtained in the above step was added with stirring and stirred for 12 hours. The reaction mixture was concentrated, ethyl acetate and water were added to the concentrate, the pH of the aqueous phase was adjusted with a saturated ammonium chloride solution, and the organic phase was washed with a saturated saline solution, dried, filtered, concentrated, and sample-mixed, and eluted with petroleum ether and ethyl acetate (2).

IIIB-18-1 (300 mg) was added to the flask in sequence, 2M HCl (diluted from 4M HCl in methanol in THF) was added and stirred at room temperature for 16 h, TLC checked for completion of the starting material reaction. Concentrating, and separating by thin layer chromatography to obtain oily compound IIIB-18 200mg. ¹ H NMR(600MHz,DMSO-d6)δ10.56(s,1H),8.63(d,J＝9.2Hz,1H),8.54(s,2H),8.46(d,J＝2.7Hz,1H),8.31(dd,J＝9.2,2.7Hz,1H),7.97(d,J＝2.7Hz,1H),7.74(dd,J＝8.9,2.8Hz,1H),7.47(d,J＝8.9Hz,1H),6.05(s,2H),4.19(t,J＝6.3Hz,2H),2.95(t,J＝6.8Hz,2H),2.57–2.54(m,3H),1.99–1.95(m,2H).ESI-MS m/z 495.2(M+H) ^- 。

Pharmacological experiments:

test experiment for anti-neocoronaviruses (SRAS-CoV-2) activity

The experimental cells were added at 8 ten thousand per wellCell density of (2) was inoculated into a 24-well cell culture plate, and 5% of CO ₂ And cultured overnight in an incubator at 37 ℃. The next day compound (1 μ M or 10 μ M, double wells) and SRAS-CoV-2 (infection at MOI = 0.01) were added. In 5% of CO ₂ After culturing at 37 ℃ in an incubator for 1 day, the supernatant was collected and the number of RNA copies of the supernatant was determined. If the copy number of the supernatant virus RNA of the wells containing the compound is obviously lower than that of the group without the compound, the compound has an inhibition effect on the tested SRAS-CoV-2.

The anti-SRAS-CoV-2 activity of the compound was expressed by the inhibition (%) of the RNA copy number of the virus supernatant by the compound, as shown in tables 1 and 2. The calculation formula is as follows: inhibition (%) = 1-number of copies of viral RNA in Compound-free group/number of copies of viral RNA in Compound-free group X100%

TABLE 1.10 inhibition of the copy number of the RNA of the novel coronavirus at μ M

Test compounds	10 μ M inhibition (%)
		IIIA-1	N/A
IIIA-2	＊
		IIIA-3	＊＊＊＊
IIIA-4	＊＊＊＊
		IIIA-5	N/A
IIIA-6	＊＊＊＊
		IIIA-7	＊＊
IIIA-8	N/A
		IIIA-20	＊＊＊＊
IIIA-21	＊
		IIIA-22	＊＊＊
IIIA-29	＊＊＊

TABLE 2.1 inhibition of the copy number of the RNA of the new coronavirus at μ M

Description of the drawings: N/A: no inhibition; it is: 1 to 25; it is a new method for preparing the compound: 26-50; twining: 51-75; twining: 76-100

anti-Valley fever Virus (RVFV) and fever-associated-thrombocytopenia syndrome Virus (SFTSV) Activity assays

Vero cells were plated at 1X10 ⁴ Perwell inoculation into 96 well cell culture plates at 5% CO ₂ And cultured overnight in an incubator at 37 ℃. The following day, different concentrations of compound (see table 1) were added, 2 replicates per concentration gradient. Equal volumes of DMSO were used as negative controls and 10 μ M Benidipine as positive control. (ii) 1h after pretreatment of the compounds, addition of RVFV-eGFP, SFTSV infection (MOI = 0.5) to 5% CO ₂ And continuously culturing for 24 hours in an incubator at 37 ℃. Cells were fixed with 4% paraformaldehyde 24h after viral infection. After the RVFV-eGFP infected group is fixed, the cell nucleus is stained by a DAPI dye (1. After fixation of the SFTSV-infected panel, specific NP antibodies (1, 1h at room temperature or overnight at 4 ℃) and fluorescent secondary antibodies (1, 1000, 1h away from light at room temperature) were incubated, followed by staining of nuclei with DAPI dye (1, 2000, 10min away from light at room temperature), and finally, detection was performed using an Operetta high content drug screening system. Nuclei were detected using 405 channels and the lower number of nuclei stained with DAPI compared to the DMSO group indicated that the compound was cytotoxic at this concentration. Viral signals were detected using the 488 channel and the lower number of cells detected compared to the DMSO group indicated that the compound had an antiviral effect at this concentration.

The antiviral activity of the compounds is represented by the inhibition rate (%) of the compounds against viral replication, as shown in tables 3 and 4. The calculation formula is as follows: inhibition (%) = (DMSO group mean reading-drug well mean reading)/DMSO group mean reading × 100.

TABLE 3 inhibition of RVFV by different concentrations of the compound

Test compounds	1 μ M inhibition (%)	0.1. Mu.M inhibition (%)	0.01. Mu.M inhibition (%)
				IIIA-4	＊＊＊＊	＊	＊
IIIA-9	＊＊＊＊	＊	＊
				IIIA-14	＊＊＊＊	＊	＊
IIIA-18	＊＊＊＊	＊	＊
				Niclosamide	＊＊＊	＊	＊

Description of the drawings: N/A: no inhibition; it is: 1 to 25; it is, zhu: 26-50; twining: 51-75; twining the processes: 76-100

TABLE 4 inhibition of SFTSV by various concentrations of Compounds

Test compounds	1 μ M inhibition (%)	0.1. Mu.M inhibition (%)	0.01. Mu.M inhibition (%)
				IIIA-4	＊＊＊	＊	＊
IIIA-9	＊＊＊＊	＊	＊
				IIIA-14	＊＊＊	＊	＊
IIIA-18	＊＊＊＊	＊	＊
				IIIA-25	N/A	N/A	N/A
IIIA-27	＊＊	N/A	N/A
				IIIA-12	＊＊	N/A	N/A
IIIA-13	＊	N/A	N/A
				IIIA-16	＊＊	N/A	N/A
IIIA-17	＊＊＊＊	＊	＊
				IIIA-30	＊＊＊＊	＊	＊
IIIA-31	＊	N/A	N/A
				IIIA-32	＊＊	N/A	N/A
IIIA-33	＊＊＊	＊	N/A
				IIIB-1	＊	N/A	N/A
IIIB-3	＊＊＊＊	＊	＊
				IIIB-6	＊＊＊＊	＊	＊
IIIB-7	＊	N/A	N/A
				IIIB-4	＊	N/A	N/A
IIIB-8	＊＊＊	＊	N/A
				IIIB-9	＊	N/A	N/A
IIIB-5	＊＊＊	＊	N/A
				IIIB-10	＊	N/A	N/A
IIIB-11	＊	N/A	N/A
				IIIB-12	＊	N/A	N/A
IIIB-13	＊	N/A	N/A
				IIIB-14	＊	N/A	N/A
Niclosamide	＊＊＊	＊	＊

Description of the invention: N/A: no inhibition; it is: 1 to 25; it is, zhu: 26-50; twining: 51-75; twining: 76-100

Test for anti-dengue Virus Activity

Inoculating Vero cells into a 6-well cell culture plate at a density of 600,000 cells per well, in 5% CO ₂ And cultured overnight in an incubator at 37 ℃. The next day compounds (1-5 concentration points, single spots) and viruses (40-50 PFU/well) were added. The cells were cultured in an incubator at 37 ℃ for 2 hours under the condition of 5% CO2, followed by blotting the supernatant and adding a low melting point agarose medium containing the corresponding concentration of the compound. The cells were incubated in an incubator at 5% CO2, 33 ℃ or 37 ℃ for 6-7 days until significant viral plaques could be observed microscopically in virus-free control wells. Cells were fixed using 4% paraformaldehyde and stained with crystal violet (see table 5). The number of plaques in each well was counted.

Cytotoxicity experiments were performed in parallel with antiviral experiments. Vero cells were seeded at a density of 20,000 cells per well in 96-well cell culture plates at 5% CO ₂ And cultured overnight in an incubator at 37 ℃. The next day compound (1-5 concentration points, single spots) was added. The cells were cultured in an incubator at 5% CO2, 33 ℃ or 37 ℃ for 6-7 days. Cell viability was then measured for each well using CCK-8.

The antiviral activity and cytotoxicity of the compounds were represented by the inhibition rate (%) of viral plaque and the cell viability (%) of the compounds, respectively, see table 6. The calculation formula is as follows:

inhibition (%) = 100-number of test well plaques/number of virus control well plaques × 100

Cell viability (%) = (test well reading-medium control mean)/(cell control mean-medium control mean) × 100

EC ₅₀ And CC ₅₀ Values were calculated by Prism software (version 5) and the inhibition curve fitting method was "log (inhibitor) vs. response- -Variable slope".

TABLE 5 anti-dengue virus Activity test treatment protocol

TABLE 6 test results for anti-dengue Virus activity

Test compounds	1 μ M inhibition (%)
		Niclosamide	＊＊＊
IIIA-4	＊＊＊＊
		IIIA-6	＊＊＊＊
IIIA-10	＊＊＊＊
		IIIA-21	N/A

Description of the drawings: N/A: no inhibition; it is: 1 to 25; it is, zhu: 26-50; tprovides a method for producing a compound: 51-75; twining: 76-100

Test experiment for interleukin 6 secretion inhibiting activity

Cells were seeded at a density of 50000 cells per well in 48 well cell culture plates at 5% CO ₂ And cultured overnight in an incubator at 37 ℃. The next day the compound (1 concentration point, duplicate wells) was added, the cytokine interleukin 6 was added after 20 minutes, at 5% CO ₂ And the culture box is kept for 8 hours at 37 ℃, and the cells are collected and the reporter gene is detected after 8 hours. The inhibition results are shown in table 7.

TABLE 7 results of the Interleukin 6 secretion inhibiting Activity test

Test compounds	1 μ M inhibition (%)
		Niclosamide	＊＊＊
IIIA-4	＊＊
		IIIA-3	＊＊
IIIA-2	＊
		IIIA-9	＊＊＊
IIIA-24	＊
		IIIA-15	＊
IIIA-26	＊
		IIIA-18	＊＊＊

Description of the invention: N/A: no inhibition; it is: 1 to 25; it is a new method for preparing the compound: 26-50; tprovides a method for producing a compound: 51-75; twining the processes: 76-100

Test experiment for interleukin 33 secretion inhibiting activity

The test method is the same as the test for inhibiting the activity of interleukin 6 except that the cytokine interleukin 6 is changed into the cytokine interleukin 33. The inhibition results are shown in table 8.

TABLE 8 test results of interleukin 33 secretion inhibiting activity

Test compounds	1 μ M inhibition (%)	5 μ M inhibition (%)
			Niclosamide	＊＊	＊＊＊
IIIA-4	＊＊	＊＊
			IIIA-9	N/A	N/A
IIIA-15	＊＊	＊＊＊
			IIIA-18	＊＊＊＊	＊＊＊＊
IIIA-14	＊	＊＊
			IIIA-12	＊＊＊＊	＊＊＊＊
IIIA-30	＊＊＊＊	＊＊＊
			IIIA-31	N/A	＊
IIIA-32	＊＊＊	＊＊＊＊

Test for Interleukin 1b secretion inhibiting Activity

The test method is the same as the test for inhibiting the activity of interleukin 6 except that the cytokine interleukin 6 is changed into the cytokine interleukin 1 b. The inhibition results are shown in table 9.

TABLE 9 results of measurement of interleukin 1b secretion inhibiting activity

Description of the invention: N/A: no inhibition; it is: 1 to 25; it is a new method for preparing the compound: 26-50; twining: 51-75; twining the processes: 76-100

Test experiment for TNF-alpha activity inhibition

The test method is the same as the test for inhibiting the activity of interleukin 6 except that the cytokine interleukin 6 is changed into the cytokine interleukin TNF-alpha. The inhibition results are shown in table 10.

TABLE 10 test results for TNF-alpha inhibitory activity

Test compounds	1 μ M inhibition (%)	5 μ M inhibition (%)
			Niclosamide	＊	＊＊
IIIA-4	＊	＊＊＊＊
			IIIA-9	N/A	N/A
IIIA-15	＊＊	＊＊
			IIIA-18	＊＊＊	＊＊＊＊
IIIA-14	＊＊	＊＊＊
			IIIA-12	＊＊＊＊	＊＊＊＊
IIIA-30	＊＊＊	＊＊＊＊
			IIIA-31	N/A	N/A
IIIA-32	＊＊	＊＊

Description of the drawings: N/A: no inhibition; it is: 1 to 25; it is a new method for preparing the compound: 26-50; tprovides a method for producing a compound: 51-75; twining: 76-100

As can be seen from the data in tables 1-10 above, the compounds of the examples of the present application are superior to niclosamide in the activity against novel coronaviruses, rift valley fever virus and fever with thrombocytopenia syndrome virus, and inhibit the secretion of interleukin-6 induced by LPS, indicating that some of the compounds of the examples of the present application have broad-spectrum antiviral activity and anti-inflammatory activity, and can be developed into antiviral drugs or anti-inflammatory drugs with potential.

Claims

1. An amide compound represented by the following general formula I, a prodrug, a pharmaceutically acceptable salt, a complex or a solvate thereof:

wherein, the first and the second end of the pipe are connected with each other,

y is a phenyl ring, a 5-6 membered heteroaromatic ring, a phenyl and 5-6 membered heteroaromatic ring or a 5-6 membered heteroaromatic and 5-6 membered heteroaromatic ring;

z is O or S;

A ₁ is C-OR ₁₁ 、C-SR ₁₁ 、C-NHR ₁₁ N or CR ₁₂ ；

C1-C20 alkylphosphoryl, C1-C20 alkylsulfonyl, C1-C20 alkyl substituted 6-20 member heterocyclylcarbonyl or C1-C20 alkyl substituted heterocyclylcarbonyl containing 1-3 hetero atoms selected from O, N or S

The C1-C20 alkyl is optionally selected from hydroxy, -NH ₂ C1-C20 alkylamino, cyano, carboxyl, halogen,

C1-C10 alkyl sulfonyl, C1-C10 alkyl phosphoryl and C1-C10 acyl;

R ₇ is C1-C10 alkyl, C1-C10 alkylamino C1-C10 alkyl or C3-C10 cycloalkyl;

R ₈ is hydrogen atom, halogen or C1-C10 alkyl;

R ₁₂ is hydrogen atom, halogen, cyano, nitro or C1-C20 alkyl;

A ₂ 、A ₃ 、A ₄ 、A ₅ each independently is N or CR ₂ ；

R ₃ is a hydrogen atom or a C1-C6 alkyl group;

R ₄ to R ₆ Each independently selected from the group consisting of hydrogen, hydroxy, amino, halogen, nitro, cyano, C1-C6 alkyl, C1-C6 alkyloxy and C1-C6 alkylthio, said C1-C6 alkyl being optionally substituted by one or more of hydroxy, amino, cyano, carboxy, halogen, C1-C6 alkylsulfonyl, C1-C6 alkylphosphoryl, C1-C6 alkylacyl.

2. The amide-based compound according to claim 1, a prodrug, a pharmaceutically acceptable salt, a complex, or a solvate thereof,

y is a benzene ring, a pyridine ring or a thiazole ring;

R ₁₁ is hydrogen atom, C1-C10 alkyl, C1-C10 alkylcarbonyl, C1-C10 alkylaminocarbonyloxy, C1-C10 alkyl-substituted 6-to 10-membered heterocyclylcarbonyl containing 1 to 3 heteroatoms selected from O, N or S or

The C1-C10 alkyl is optionally substituted by a substituent selected from the group consisting of hydroxy, amino, cyano, carboxy, halogen,

C1-C10 alkylsulfonyl, C1-C10 alkylphosphoryl and C1-C10 alkylcarbonyl;

R ₇ is C1-C6 alkyl, C1-C6 alkylamino C1-C6 alkylOr a C3-C7 cycloalkyl group;

R ₈ is hydrogen atom, halogen or C1-C2 alkyl;

R ₁₂ is hydrogen atom, halogen, cyano, nitro or C1-C6 alkyl;

R ₂ are the same or different from each other and are each independently selected from the group consisting of hydrogen, hydroxy, halogen, C1-C4 alkyloxy, C1-C4 alkyl, amino, nitro, cyano and C1-C4 alkylcarbonyl, said C1-C4 alkyl being optionally substituted by one or more of hydroxy, amino, cyano, carboxy, halogen, C1-C4 alkylsulfonyl, C1-C4 alkylphosphoryl, C1-C4 alkylcarbonyl;

R ₃ is a hydrogen atom or a C1-C4 alkyl group;

R ₄ to R ₆ Each independently selected from hydrogen, hydroxy, amino, halogen, nitro, cyano, C1-C4 alkyl and C1-C4 alkyloxy, the C1-C4 alkyl being optionally substituted by one or more of hydroxy, amino, cyano, carboxy, halogen.

3. The amide-based compound according to claim 2, a prodrug, a pharmaceutically acceptable salt, a complex, or a solvate thereof,

R ₁₁ is a hydrogen atom, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, an aminoC 1-C6 alkyl group, a hydroxyl group, a C1-C6 alkyl group, a C1-C6 alkylcarbonyl group, a C1-C6 alkylaminocarbonyl group, a C1-C6 alkylcarbonyloxy C1-C6 alkyl group, a C1-C6 alkylaminoC 1-C6 alkylcarbonyl group, an aminoC 1-C6 alkylcarbonyl group, a,

Substituted C1-C6 alkylcarbonyl, C1-C6 alkyl-substituted 6-to 8-membered heterocyclylcarbonyl containing 1 to 3 heteroatoms from the group consisting of O, N or S or

R ₇ Is C1-C4 alkyl, C1-C3 alkylamino C1-C4 alkyl or C3-C6 cycloalkyl;

R ₈ is a hydrogen atom or a methyl group;

R ₁₂ is hydrogen atom, halogen, cyano or C1-C4 alkyl;

A ₂ 、A ₃ 、A ₄ 、A ₅ at most two of N;

R ₂ are identical or different from one another and are each independently selected from the group consisting of hydrogen, hydroxy, halogen, C1-C2 alkoxy, C1-C2 alkyl, C1-C2 alkylcarbonyl, haloC 1-C2 alkoxy, haloC 1-C2 alkyl, haloC 1-C2 alkylcarbonyl, amino and nitro;

R ₃ is a hydrogen atom;

R ₄ to R ₆ Each independently selected from a hydrogen atom, a halogen, a C1-C4 alkyl group, a halogenated C1-C4 alkyl group, a C1-C4 alkoxy group, and a halogenated C1-C4 alkoxy group.

4. The amide-based compound according to claim 3, a prodrug, a pharmaceutically acceptable salt, a complex, or a solvate thereof,

R ₁₁ is a hydrogen atom, a C1-C4 alkyl group, a C1-C6 alkylcarbonyl group, a C1-C5 alkylaminocarbonyl group, a C1-C5 alkylcarbonyloxy C1-C5 alkyl group,

Substituted C1-C4 alkyl a substituted C1-C4 alkylcarbonyl group,

Or

5. The amide-based compound, a prodrug thereof, a pharmaceutically acceptable salt, a complex or a solvate thereof according to claim 1, wherein the amide-based compound represented by the general formula I is a compound represented by the following general formula II, IIIA or IIIB:

wherein, A ₁ Is C-OR ₁₁ 、C-SR ₁₁ 、C-NHR ₁₁ Or CR ₁₂ ；

R ₁₁ 、R ₁₂ 、R ₂ 、R ₄ To R ₆ And Y is as defined in claim 1.

6. The amide-based compound, a prodrug thereof, a pharmaceutically acceptable salt, a complex or a solvate thereof according to claim 1, wherein the amide-based compound represented by the general formula I is a compound represented by the following general formula IV:

wherein R is ₁₁ 、R ₂ 、R ₄ And R ₅ Is as defined in claim 1.

7. The amide-based compound, a prodrug, a pharmaceutically acceptable salt, a complex, or a solvate thereof according to claim 1, wherein the compound or salt is selected from the following structures:

8. a pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the group consisting of a compound according to any one of claims 1 to 7, a prodrug, a pharmaceutically acceptable salt, a complex and a solvate thereof, and optionally a pharmaceutically acceptable carrier.

9. Use of a compound according to any one of claims 1 to 7, prodrugs, pharmaceutically acceptable salts, complexes and solvates thereof, or a pharmaceutical composition according to claim 8, in the manufacture of an antiviral medicament or a medicament for the treatment of an anti-inflammatory disease.

10. The use according to claim 9, wherein the virus is at least one selected from the group consisting of a DNA virus, an RNA virus;

in particular, the DNA virus is selected from the group consisting of hepatitis b virus, human papilloma virus, herpes virus, bacteriophage virus; the RNA virus is selected from influenza virus, avian influenza virus, zika virus, dengue fever virus, respiratory syncytial virus, SARS virus, MERS virus, novel coronavirus (SARS-CoV-2), bunyavirus, AIDS virus, ebola virus, hepatitis C virus, encephalitis B virus, rhinovirus, poliovirus, coxsackie virus, rotavirus, tobacco mosaic virus, bacteriophage virus, marburg virus and enterovirus,

in particular, the RNA virus is at least one of coronavirus, bunyavirus and dengue virus;

in particular, the coronavirus is SARS-CoV-2; the bunyavirus is rift valley fever virus;

more particularly, the virus is dengue virus;

the inflammatory diseases are inflammatory skin diseases, inflammatory bowel diseases, inflammatory joint diseases and other autoimmune diseases.