CN106957290B - A kind of aromatic carboxamide compound and its application in preparing anti-influenza A virus medicine - Google Patents

Abstract

The invention discloses an aromatic carboxamide compound and its application in preparing an anti-influenza A virus drug, belonging to the technical field of medicine. Specifically, using aromatic heterocyclic carboxylic acid and 2-(benzylthio)ethylamine derivatives as raw materials, a series of aromatic carboxamides were prepared by amide reaction. The aromatic carboxamide compounds and pharmaceutically acceptable salts thereof can effectively inhibit the activity of influenza A virus, and have low toxicity to cells, showing the application prospect of such compounds in the preparation of anti-influenza A virus drugs.

Description

A kind of aromatic carboxamide compound and its preparation anti-influenza A virus medicine Applications

technical field

The invention belongs to the technical field of medicine, and relates to an aromatic carboxamide compound and its application in preparing an anti-influenza A virus medicine.

Background technique

Influenza (hereinafter referred to as influenza) is a serious upper respiratory tract disease caused by influenza virus. Influenza viruses can cause influenza in various organisms such as humans, pigs, horses, dogs and birds. Influenza viruses are RNA viruses that belong to the family Orthomyxoviridae. Influenza viruses can be divided into three subtypes based on their nucleoproteins and matrix proteins: A, B, and C. Influenza A virus (IAV) can be classified into influenza A virus according to the antigenic difference of two surface glycoproteins, Hemagglutinin (HA) and Neuraminidase (NA). For a variety of subtypes, such as H1N1, H2N2, H5N1 subtype and so on. The H1N1 influenza in Spain in 1918 killed more than 20 million people, and the H5N1 influenza in 2005 caused hundreds of deaths, and the mortality rate was as high as 60%. In 2013, the first H7N9 case was discovered in my country; according to the National Health and Family Planning Commission's Bureau of Disease Control and Prevention, 79 people died in January 2017 due to avian influenza H7N9 infection. In addition, every year avian influenza (H5N1, H7N9, etc.) has caused huge economic losses to my country's poultry industry.

At present, there are two main types of drugs for the clinical treatment of influenza: M2 ion channel inhibitors, which inhibit the uncoating process of the virus by preventing the acidification of the intracellular virus. The FDA-approved drugs include amantadine and amantadine; Neuraminidase inhibitors have anti-influenza virus effects by inhibiting the release of progeny viruses from host cells. The FDA-approved drugs include oseltamivir and zanamivir.

In recent years, the frequent emergence of resistant mutants of M2 ion channel inhibitors and neuraminidase inhibitors makes it particularly important to develop anti-influenza drugs with new structural backbones or new targets.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a class of aromatic carboxamide compounds represented by general formula I or a pharmacologically or physiologically acceptable salt thereof,

in,

R ¹ is H, 4-F, 4-Cl, 4-Br, 4-CN, 4-CF ₃ , 4-CH ₃ , 4-OCH ₃ , 2-NO ₂ , 3-NO ₂ , 4-NO ₂ or 3-Cl;

^R2 is

X is C or S; n is 0 or 3.

Preferably, the present invention provides the compounds shown in Table 1 below:

The present invention finds that the above-mentioned aromatic carboxamide compounds can be used to prepare anti-influenza A virus drugs through in vitro anti-influenza A virus activity experiments.

More preferably, especially the following compounds:

N-(2-(benzylthio)ethyl)-2,5-dimethylfuran-3-carboxamide,

N-(2-((4-Chlorobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide,

N-(2-((4-cyanobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide,

N-(2-((3-nitrobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide,

N-(2-((4-nitrobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide,

N-(2-((4-Chlorobenzyl)thio)ethyl)-2,5-dimethylthiophene-3-carboxamide,

N-(2-((4-fluorobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide,

N-(2-((4-Bromobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide,

2,5-Dimethyl-N-(2-((4-nitrobenzyl)thio)ethyl)thiophene-3-carboxamide,

N-(2-((4-cyanobenzyl)thio)ethyl)-2,5-dimethylthiophene-3-carboxamide,

N-benzyl 2,5-dimethylfuran-3-carboxamide.

The present invention also provides a method for preparing the aromatic carboxamide compounds represented by the above structural formula.

The aromatic heterocyclic carboxylic acid derivative with R is synthesized by the reaction shown in the following formula i. The specific operation steps can be as follows:

Calcium hypochlorite (3.1 g, 21.71 mmol) was dissolved in 6 mL of warm water, and then 3 mL of warm aqueous solution dissolved with potassium carbonate (2.6 g, 18.82 mmol) and potassium hydroxide (727.5 mg, 30.39 mmol) was slowly added dropwise. Add to the above solution and stir well until the semi-solid gel becomes liquid. The suspension was filtered and the solid was washed with 10 mL to obtain a filtrate, and the filtrate was heated to 55° C. Then 3-acetyl-2,5-dimethylfuran (5d) (1 g, 7.237 mmol) was slowly added dropwise while stirring, and the addition was completed. , the reaction temperature was kept at 60-70 °C. After 1 hour of reaction, the reaction was monitored by TLC. After the reaction was completed, sodium sulfite was added to remove excess hypochlorous acid, the solution was cooled and acidified, then extracted with ethyl acetate (3×30 mL), dried with saturated brine and anhydrous sodium sulfate, and concentrated. After separation by column chromatography, 2,5-dimethyl-3-furanic acid (6d) was obtained as the raw material of reaction iv. The preparation methods of other aromatic heterocyclic carboxylic acid derivatives with R are the same as above.

1-(Bromomethyl)-2-nitrobenzene (2a) and 1-(bromomethyl)-3-nitrobenzene (2b) were synthesized by the reaction shown in the following formula ii. The specific operation steps can be as follows:

PBr ₃ (494.8 mg, 1.828 mmol) was slowly added dropwise to a solution of (2-nitrophenyl) methanol or (3-nitrophenyl) methanol (400 mg, 2.612 mmol) in ether under ice bath conditions, The ice bath is then stirred for 2 to 4 hours. The reaction was monitored by TLC, the reaction was completed, the reaction was quenched with 10 mL of water and extracted with ethyl acetate (3×30 mL), dried with saturated brine and anhydrous sodium sulfate, concentrated and separated by column chromatography to obtain 1-(bromomethyl)-2 - Nitrobenzene (2a) or 1-(bromomethyl)-3-nitrobenzene (2b) as starting material for reaction iii.

The 2-(benzylthio)ethylamine derivative with R ¹ is synthesized by the reaction shown in the following formula iii. The specific operation steps can be as follows:

Lithium hydroxide (105.4 mg, 4.40 mmol) was dissolved in 3 mL of water, then 9 mL of ethanol was added, the above solution was added dropwise to a 25 mL round-bottomed flask containing cysteamine hydrochloride (200 mg, 1.76 mmol), and finally Benzyl chloride (234.2 mg, 1.85 mmol) was added, and the mixture was stirred in an oil bath at 35°C for 40 min. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was concentrated, then extracted with ethyl acetate (3×30 mL), dried with saturated brine and anhydrous sodium sulfate, and concentrated by column chromatography to obtain 2-(benzylthio)ethylamine (2i ) as the starting material for reaction iv. The preparation methods of other 2-(benzylthio)ethylamine derivatives with R ¹ are the same as above.

Through the synthesis, the aromatic heterocyclic carboxylic acid derivative with R was obtained, and then 1.05 equivalents of the 2-(benzylthio)ethylamine derivative with R ¹ , HOBT and EDCI were placed in a 5 mL round-bottomed flask, and argon was passed through. Finally, 3 mL of DMF was added to react overnight at room temperature, and the reaction was monitored by TLC. After the reaction was completed, the target compound was separated by column chromatography. The reaction formula is shown in the following formula iv.

in,

R ¹ is H, 4-F, 4-Cl, 4-Br, 4-CN, 4-CF ₃ , 4-CH ₃ , 4-OCH ₃ , 2-NO ₂ , 3-NO ₂ , 4-NO ₂ or 3-Cl;

^R2 is

X is C or S; n is 0 or 3.

The specific operation process can be as follows:

Take 0.4 mmol of the carboxylic acid derivative of reaction i and 0.42 mmol (1.05 eq.) of the 2-(benzylthio)ethylamine derivative of reaction iii with R ¹ and HOBT (0.4 mmol) and EDCI (0.4 mmol) were added to Into a 5mL single-neck round-bottomed flask containing a magneton, argon was introduced, and finally 3mL of DMF was added to dissolve it, stirred at room temperature overnight, and TLC monitored the reaction. compound.

Another object of the present invention is to provide a pharmaceutical composition against influenza A virus, comprising the above aromatic carboxamide compounds or their pharmacologically or physiologically acceptable salts and a pharmaceutically acceptable carrier or excipient. .

The aromatic carboxamide compounds involved in the present invention can effectively inhibit the activity of influenza A virus, have low toxicity to cells, and can be used for preparing medicines against influenza A virus.

Detailed ways

The features and advantages of the present invention can be further understood from the following detailed description. The examples provided are merely illustrative of the methods of the present invention, and are not intended to limit the remainder of the present disclosure in any way.

[Example 1]: Preparation of N-(2-(benzylthio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS1)

Take 2,5-dimethyl-3-furanic acid (56 mg, 0.4 mmol), 2-(benzylthio)ethan-1-amine (70.3 mg, 0.42 mmol), HOBT (54 mg, 0.4 mmol) and EDCI ( 76.7mg, 0.4mmol), was added to the 5mL single-necked round-bottomed flask containing the magneton, passed into argon, and finally added 3mL of DMF to dissolve it, stirred at room temperature overnight, and TLC monitored the reaction. The pure target compound YYS-1 was obtained by analytical separation, and the product was a colorless liquid with a yield of 78%. ¹ HNMR (400MHz, CDCl ₃ ) δ 7.32-7.27(m, 4H), 7.26-7.20(m, 1H), 6.19(s, 1H), 6.00(d, J=0.7Hz, 1H), 3.72(s) , 2H), 3.49(dd, J=12.5, 6.2Hz, 2H), 2.62(t, J=6.4Hz, 2H), 2.52(s, 3H), 2.22(s, 3H). ¹³ C NMR(100MHz, CDCl ₃ )δ164.19,155.09,149.92,138.09,128.86,128.62,127.17,115.95,104.14,37.75,35.76,31.22,13.51,13.30.HRMS(ESI)calcd for C ₁₆ H ₁₉ NO ₂ S[M ⁺ H] 290.1215, found 290.1208.

[Example 2]: Preparation of N-(2-((3-chlorobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS2)

The preparation method was as in Example 1, and the product was a colorless liquid with a yield of 39%. ¹ H NMR (400MHz, CDCl ₃ )δ7.33(s,1H), 7.26-7.15(m,3H), 6.11(s,1H), 5.99(s,1H), 3.69(s,2H), 3.51( q, J=6.2Hz, 2H), 2.64 (t, J=6.4Hz, 2H), 2.53 (d, J=2.8Hz, 3H), 2.24 (s, 3H). ¹³ C NMR (100MHz, CDCl ₃ ) δ164.24,155.25,150.02,140.22,134.41,129.84,128.90,127.39,127.03,115.80,104.01,37.73,35.30,31.33 ₊ 13.51,13.29.HRMS(ESI) calcd for C ₁₆ H ₁₈ ⁺ 324.0825, found 324.0818.

[Example 3]: Preparation of N-(2-((4-chlorobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS3)

The preparation method is as in Example 1, the product is a colorless liquid, and the yield is 11%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.31-7.21 (m, 4H), 6.07 (s, 1H), 5.98 (d, J=0.5Hz, 1H), 3.69 (s, 2H), 3.51 (dd, J=12.4, 6.3Hz, 2H), 2.62 (t, J=6.4Hz, 2H), 2.53 (s, 3H), 2.24 (s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 164.19, 155.22, 150.04 ,136.60,132.93,130.20,128.76,115.79,103.98,37.72,35.09,31.19,13.51,13.31.HRMS(ESI)calcd for C ₁₆ H ₁₈ ClNO ₂ S[M+H] ⁺ 324.0825,found 324.0818.

[Example 4]: Preparation of N-(2-((4-cyanobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS4)

The preparation method was as in Example 1, and the product was a colorless liquid with a yield of 56%. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.66-7.55(m, 2H), 7.45(d, J=8.4Hz, 2H), 6.12(s, 1H), 5.99(s, 1H), 3.77(s, 2H), 3.52(q, J=6.5Hz, 2H), 2.62(t, J=6.6Hz, 2H), 2.53(s, 3H), 2.24(s, 3H). ¹³ C NMR (100MHz, CDCl ₃ ) δ164.24,155.29,150.07,143.90,132.40,129.65,118.75,115.73,110.91,103.94,37.89,35.54,31.28,13.50,13.30.HRMS(ESI)calcd for C ₁₇ H ₁₈ N ₂ O ₂ S[M+H 18 N 2 O 2 S ⁺ 315.1167,found315.1160.

[Example 5]: Preparation of N-(2-((3-nitrobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS5)

The preparation method is as in Example 1, the product is a white solid, the yield is 47%, and the melting point is 113-115°C. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.21 (d, J=1.5 Hz, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.67 (d, J=7.5 Hz, 1H), 7.56-7.43 (m, 1H), 6.28–6.13 (m, 1H), 6.01 (s, 1H), 3.83 (s, 2H), 3.54 (q, J=6.2Hz, 2H), 2.65 (t, J=6.4Hz, 2H), 2.53(s, 3H), 2.23(s, 3H). ¹³ C NMR (100MHz, CDCl ₃ )δ164.27, 155.31, 150.06, 148.33, 140.49, 135.07, 129.53, 123.70, 122.22, 115.71, 103.95, 37.9 35.23, 31.38, 13.48, 13.26. HRMS(ESI) calcd for C ₁₆ H ₁₈ N ₂ O ₄ S[M+H] ⁺ 335.1066, found 335.1057.

[Example 6]: Preparation of N-(2-((4-nitrobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS6)

The preparation method is as in Example 1, the product is a white solid, the yield is 19%, and the melting point is 105-106°C. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.19-8.15 (m, 2H), 7.51 (d, J=8.7 Hz, 2H), 6.72 (d, J=0.8 Hz, 1H), 6.10 (s, 1H) , 3.82(s, 2H), 3.55(q, J=6.4Hz, 2H), 2.65(t, J=6.5Hz, 2H), 2.63(s, 3H), 2.39(s, 3H). ¹³ C NMR( 100MHz, CDCl ₃ )δ164.60,147.03,145.95,142.80,136.18,131.01,129.74,123.99,123.88,38.02,35.24,31.34,15.03,14.87.HRMS(ESI)calcd for C ₁₆ H ₁₈ N ₂ O ₄ S[ M H 18 N 2 O +H] ⁺ 335.1066,found335.1058.

[Example 7]: Preparation of N-(2-((4-chlorobenzyl)thio)ethyl)-2,5-dimethylthiophene-3-carboxamide (YYS7)

The preparation method is as in Example 1, the product is a colorless liquid, and the yield is 29%. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.35-7.14(m, 4H), 6.72(d, J=0.9Hz, 1H), 6.15(s, 1H), 3.69(s, 2H), 3.52(dd, J=12.5, 6.1Hz, 2H), 2.66-2.60(m, 5H), 2.39(s, 3H). ¹³ C NMR (100, CDCl ₃ ) δ 164.62, 142.59, 136.59, 136.04, 132.95, 131.22, 130.21, 128.77 ,124.15,37.90,35.06,31.17,15.04,14.85.HRMS(ESI)calcd for C ₁₆ H ₁₈ ClNOS ₂ [M+H] ⁺ 340.0597,found 340.0590.

[Example 8]: Preparation of N-(2-((4-fluorobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS8)

The preparation method is as in Example 1, the product is a yellow solid, the yield is 21%, and the melting point is 79-80°C. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.35-7.22 (m, 2H), 7.02-6.95 (m, 2H), 6.73 (d, J=0.9Hz, 1H), 6.18 (s, 1H), 3.71 ( s, 2H), 3.53 (dd, J=12.5, 6.1 Hz, 2H), 2.68–2.60 (m, 5H), 2.38 (s, 3H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 164.61, 142.57, 136.03, 133.77,133.74,131.23,130.44,130.36,124.15,115.59,115.38,37.88,34.98,31.16,15.02,14.84.HRMS(ESI)calcd forC ₁₆ H ₁₈ FNO ₂ S[M+H] ⁺ 301115.121,found

[Example 9]: Preparation of N-(2-((4-bromobenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS9)

The preparation method was as in Example 1, and the product was a colorless liquid with a yield of 37%. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.47-7.37 (m, 2H), 7.24-7.14 (m, 2H), 6.06 (s, 1H), 5.98 (d, J=0.6Hz, 1H), 3.67 ( s, 2H), 3.51(dd, J=12.5, 6.2Hz, 2H), 2.62(t, J=6.5Hz, 2H), 2.53(s, 3H), 2.24(s, 3H). ¹³ C NMR(100MHz) , _{CDCl 3 ) δ164.20,155.19,150.02,137.15,131.70,130.56,121.00,115.82,104.01,77.44,77.12,76.80,37.77,35.15,31.19,13.51,13.16} S[M+H] ⁺ 370.0299, found 370.0291.

[Example 10]: Preparation of 2,5-dimethyl-N-(2-((4-(trifluoromethyl)benzyl)thio)ethyl)furan-3-carboxamide (YYS10)

The preparation method is as in Example 1, the product is a white solid, the yield is 36%, and the melting point is 62-63°C. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.57 (d, J=8.1 Hz, 2H), 7.44 (d, J=8.0 Hz, 2H), 6.06 (s, 1H), 5.98 (s, 1H), 3.77 (s, 2H), 3.53(q, J=6.3Hz, 2H), 2.63(t, J=6.5Hz, 2H), 2.53(s, 3H), 2.24(s, 3H). ¹³ CNMR (100MHz, CDCl) ₃ )δ164.22,155.27,150.07,142.28,129.19,125.58,125.54,115.76,103.93,37.75,35.30,31.27,13.48,13.25.HRMS(ESI)calcd for C ₁₇ H ₁₈ F ₃ NO ₂ S[M+H] ⁺ 358.1089, found 358.1082.

[Example 11]: Preparation of 2,5-dimethyl-N-(2-((4-methylbenzyl)thio)ethyl)furan-3-carboxamide (YYS11)

The preparation method was as in Example 1, and the product was a colorless liquid with a yield of 61%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.20 (d, J=8.0 Hz, 2H), 7.11 (d, J=7.8 Hz, 2H), 6.12 (s, 1H), 5.99 (s, 1H), 3.69 (s, 2H), 3.49(dd, J=12.4, 6.0Hz, 2H), 2.62(t, J=6.4Hz, 2H), 2.52(s, 3H), 2.32(s, 3H), 2.24(s, 3H). ¹³ C NMR (100MHz, CDCl ₃ )δ164.16,155.10,149.94,136.84,134.94,129.32,128.74,115.92,104.10,37.64,35.12,31.18,21.12,13.50,13.31.HdRMS( _ESI7 )calc for H ₂₁ NO ₂ S[M+H] ⁺ 304.1371, found304.1364.

[Example 12]: Preparation of N-(2-((4-methoxybenzyl)thio)ethyl)-2,5-dimethylfuran-3-carboxamide (YYS12)

The preparation method is as in Example 1, the product is a white solid, the yield is 49%, and the melting point is 79-81°C. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.26-7.15(m, 2H), 6.89-6.74(m, 2H), 6.23(s, 1H), 6.00(s, 1H), 3.77(s, 3H), 3.68(s, 2H), 3.49(q, J=6.2Hz, 2H), 2.61(t, J=6.4Hz, 2H), 2.52(s, 3H), 2.22(s, 3H). ¹³ C NMR(100MHz) , _{CDCl 3} )δ164.22,158.67,155.08,149.90,129.99,129.93,115.95,113.97,104.14,55.24,37.81,35.13,31.07,13.49,13.27.HRMS(ESI)calcd for C ₁₇ H ₂₁ H] ⁺ 320.1320,found320.1311.

[Example 13]: Preparation of 2,5-dimethyl-N-(2-((2-nitrobenzyl)thio)ethyl)furan-3-carboxamide (YYS13)

The preparation method was as in Example 1, and the product was a yellow liquid with a yield of 67%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.97 (dd, J=8.1, 1.2 Hz, 1H), 7.56 (m, 1H), 7.50-7.37 (m, 2H), 6.19 (s, 1H), 6.04 ( d, J＝0.7Hz, 1H), 4.10(s, 2H), 3.52(q, J＝6.2Hz, 2H), 2.68(t, J＝6.4Hz, 2H), 2.53(d, J＝3.0Hz, 3H), 2.24(s, 3H). ¹³ C NMR (100MHz, CDCl ₃ )δ164.21, 155.19, 150.00, 148.60, 133.83, 133.24, 132.12, 128.44, 125.49, 115.81, 104.07, 37.84, 33.20, 32.3 .HRMS(ESI)calcd for C ₁₆ H ₁₈ N ₂ O ₄ S[M+H] ⁺ 335.1066, found 335.1058.

[Example 14]: Preparation of N-(2-((4-chlorobenzyl)thio)ethyl)furan-3-carboxamide (YYS14)

The preparation method is as in Example 1, the product is a white solid, the yield is 60%, and the melting point is 126-128°C. ¹ H NMR (400MHz, CDCl ₃ )δ7.93(s,1H), 7.43(s,1H), 7.32-7.22(m,4H), 6.61(s,1H), 6.39(s,1H), 3.69( s, 2H), 3.53 (q, J=6.3Hz, 2H), 2.62 (t, J=6.5Hz, 2H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 162.72, 144.84, 143.85, 136.55, 132.97, 130.20, 128.79,122.38,108.25,38.03,35.13,31.00.HRMS(ESI)calcd for C ₁₄ H ₁₄ ClNO ₂ S[M+H] ⁺ 296.0512,found 296.0505.

[Example 15]: Preparation of N-(2-((4-chlorobenzyl)thio)ethyl)furan-2-carboxamide (YYS15)

The preparation method is as in Example 1, the product is a white solid, the yield is 34%, and the melting point is 91-93°C. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.45 (dd, J=1.7, 0.8 Hz, 1H), 7.31-7.24 (m, 4H), 7.11 (dd, J=3.5, 0.8 Hz, 1H), 6.70 ( s, 1H), 6.50(dd, J=3.5, 1.8Hz, 1H), 3.72(s, 2H), 3.57(q, J=6.4Hz, 2H), 2.64(t, J=6.5Hz, 2H). ¹³ C NMR(100MHz, CDCl ₃ )δ158.41,147.76,144.02,136.55,132.95,130.22,128.77,114.40,112.22,37.75,35.17,31.02.HRMS(ESI)calcd for C ₁₄ H ₁₄ ClNO ₂ S[M+H ] ⁺ 296.0512, found 296.0505.

[Example 16]: Preparation of N-(2-((4-chlorobenzyl)thio)ethyl)-5-methylfuran-2-carboxamide (YYS16)

The preparation method was as in Example 1, and the product was a yellow liquid with a yield of 55%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.45 (dd, J=1.7, 0.8 Hz, 1H), 7.27 (s, 4H), 7.11 (dd, J=3.5, 0.8 Hz, 1H), 6.70 (s, 1H), 6.50(dd, J=3.5, 1.8Hz, 1H), 3.72(s, 2H), 3.57(q, J=6.4Hz, 2H), 2.64(t, J=6.5Hz, 2H). ¹³ C NMR(100MHz, CDCl ₃ )δ158.69,154.70,146.04,136.59,132.90,130.23,128.74,115.79,108.64,37.81,35.16,31.05,13.88.HRMS(ESI)calcd for C ₁₅ H ₁₆ ClHNO ₂ S[M ] ⁺ 310.0669, found 310.0662.

[Example 17]: Preparation of N-(2-((4-chlorobenzyl)thio)ethyl)-2-methylthiazole-4-carboxamide (YYS17)

The preparation method is as in Example 1, the product is a white solid, the yield is 61%, and the melting point is 190-191°C. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.74(t, J=5.6Hz, 1H), 8.17(s, 1H), 7.43-7.32(m, 4H), 3.78(s, 2H), 3.44- 3.37(m, 2H), 2.66(s, 3H), 2.58–2.53(m, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ169.50, 159.84, 142.63, 137.71, 134.82, 131.35, 130.67, 128.31, 38.67, 33.91, 29.89, 19.12. HRMS(ESI) calcd for C ₁₅ H ₁₇ ClN ₂ OS ₂ [M+H] ⁺ 341.0549, found 341.0542.

[Example 18]: Preparation of N-(2-((4-chlorobenzyl)thio)ethyl)-2-methylbenzamide (YYS18)

The preparation method is as in Example 1, the product is a white solid, the yield is 21%, and the melting point is 104-105°C. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.36-7.28(m, 2H), 7.25(d, J=5.5Hz, 4H), 7.19(m, 2H), 6.22(s, 1H), 3.70(s, 2H), 3.54(dd, J=12.6, 6.2Hz, 2H), 2.63(t, J=6.4Hz, 2H), 2.41(s, 3H). ¹³ C NMR (100MHz, CDCl ₃ )δ170.21, 136.59, 136.17 ,136.06,132.95,131.05,130.25,129.99,128.78,126.76,125.77,38.11,35.04,31.10,19.85.HRMS(ESI)calcd for C ₁₇ H ₁₈ ClNOS[M+H] ⁺ 320.0876,found320.086

[Example 19]: Preparation of 2,5-dimethyl-N-(2-((4-nitrobenzyl)thio)ethyl)thiophene-3-carboxamide (YYS19)

The preparation method is as in Example 1, the product is a yellow solid, the yield is 67%, and the melting point is 103-106°C. ¹ H NMR (400MHz, CDCl ₃ ) δ 8.21-8.13(m, 2H), 7.50(d, J=8.7Hz, 2H), 6.04(s, 1H), 5.97(s, 1H), 3.81(s, 2H), 3.53(q, J=6.4Hz, 2H), 2.64(t, J=6.6Hz, 2H), 2.53(s, 3H), 2.24(s, 3H). ¹³ C NMR (100MHz, CDCl ₃ ) δ164.15,155.33,150.12,147.04,145.98,129.71,123.86,115.68,103.85,37.90,35.32,31.42,13.47,13.26.HRMS(ESI)calcd for C ₁₆ H ₁₈ N ₂ O ₃ S ₂ [M ⁺ H] 351.0837, found 351.0830.

[Example 20]: Preparation of N-(2-((4-cyanobenzyl)thio)ethyl)-2,5-dimethylthiophene-3-carboxamide (YYS20)

The preparation method is as in Example 1, the product is a colorless liquid, and the yield is 64%. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.60(m, 2H), 7.45(d, J=8.2Hz, 2H), 6.05(s, 1H), 5.98(s, 1H), 3.77(s, 2H) , 3.52(q, J=6.4Hz, 2H), 2.62(t, J=6.6Hz, 2H), 2.53(s, 3H), 2.25(s, 3H). ¹³ C NMR(100MHz, CDCl ₃ )δ164. 17,155.29,150.08,143.85,132.39,129.63,118.69,115.74,110.99,103.89,37.87,35.57,31.35,13.47,13.27.HRMS(ESI)calcd for C ₁₇ H ₁₈ N ₂ OS ₂ [M+H] ⁺ 3 found 331.0932.

[Example 21]: Preparation of 2,5-dimethyl-N-(4-phenylbutyl)furan-3-carboxamide (YYS21)

The preparation method was as in Example 1, and the product was a colorless liquid with a yield of 51%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.33-7.24 (m, 2H), 7.19 (m, 3H), 6.70 (d, J=0.9 Hz, 1H), 5.77 (s, 1H), 3.39 (dd, J=12.9, 6.9Hz, 2H), 2.67–2.61 (m, 5H), 2.37 (s, 3H), 1.76–1.55 (m, 4H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 164.73, 142.13, 135.96, 131.60,128.44,128.37,125.85,124.09,39.41,35.52,29.35,28.79,15.03,14.78.HRMS(ESI) _calcd for C17H21NO2[M+H] ⁺ _272.1651 ,found _272.1642

[Example 22]: Preparation of N-benzyl 2,5-dimethylfuran-3-carboxamide (YYS22)

The preparation method is as in Example 1, the product is a white solid, the yield is 59%, and the melting point is 109-112°C. ¹ H NMR (400MHz, CDCl ₃ ) δ 7.34-7.18(m, 5H), 6.29(s, 1H), 6.02(s, 1H), 4.50(d, J=5.8Hz, 2H), 2.52(s, 3H), 2.20(s, 3H). ¹³ C NMR(100MHz, CDCl ₃ )δ164.25,155.38,149.89,138.65,128.64,127.71,127.35,115.87,104.15,43.22,13.52,13.28.HRMS(ESI)calcd for C ₁₄ H ₁₅ NO ₂ [M+H] ⁺ 30.1181, found 230.1175.

Table 1 Chemical structure of the target compound YYS1-22 synthesized by the method of the present invention

[Experimental Example 23]: Pharmacological Experiment of Aromatic Carboxamide Compounds

(1) Cytotoxicity determination of aromatic formamide compounds:

Yellow thiazolan, or MTT for short, can penetrate the cell membrane and enter the cell. The succinyl dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to insoluble blue-purple needle-like Formazan crystals and deposit in the cells. The crystals can be dissolved by 20% (mass to volume) SDS, and its light absorption value can be measured at a wavelength of 575 nm with an enzyme-linked immunosorbent assay, which can indirectly reflect the number of cells.

During the experiment, MDCK cells were transferred to a 96-well plate at a density of 2 × 10 ⁴ per well. After culturing at 37°C for 24 hours, the medium was aspirated, and cell culture medium containing various concentration gradient compounds was added to each well. hole. After 24 hours, 5 mg/mL MTT solution was added to each well, and the cell plates were incubated in a CO ₂ incubator at 37 °C for 4 h. Then, the co-solution was added to the hemolyzed cells, incubated at 37°C for 3h, and the OD value at a wavelength of 575nm was measured by a microplate reader. Compound inhibition rate (%)=[1-(EN)/(PN)]×100, where "E" represents the OD value of the administration group, "P" represents the OD value of the non-administration group, and "N" represents the OD value of the non-administration group OD value of blank group. The median inhibitory concentration (CC ₅₀ ) of the compound was used as an indicator of the cytotoxicity of the compound.

(2) In vitro anti-H5N1 activity of aromatic carboxamides:

The antiviral activity of compounds was assessed by viral plaque reduction assay. Influenza virus (H5N1) was inserted into a 6-well plate filled with MDCK cells at 70 PFU/well. After 40 minutes, the virus-containing medium was removed and a medium containing a specific concentration of the drug to be tested was added. The medium contained a final concentration of 0.001% DEAE- dextran, 2ug/ml TPCK-trypsin and 0.5% agarose. After culturing for 48-72 hours at 37°C in 5% CO ₂ , cells were fixed with 3% formalin, stained with 0.5% crystal violet, and the number of viral plaques was counted. _EC50 refers to the concentration required for a particular drug to effectively inhibit viral plaque generation by 50% of control wells.

The present invention compares with amantadine, carries out cytotoxicity and anti-influenza A virus H5N1 activity check to 22 synthetic compounds, and calculates the selectivity index SI of compounds, the results are shown in Table 2.

Table 2 The results of the anti-H5N1 activity and cytotoxicity of the target compound YYS1-22 synthesized by the present invention

Compounds EC₅₀(μM) CC₅₀(μM) SI(CC₅₀/IC₅₀) YYS 1 7.716±2.909 115.173±30.201 14.9 YYS 2 54.545±16.440 53.514±13.247 0.9 YYS 3 8.541±3.153 51.476±10.561 6.0 YYS 4 5.302±1.450 >100 >18.9 YYS 5 9.970±2.548 109.662±27.423 11.0 YYS 6 1.247±0.335 >100 >80.2 YYS 7 1.639±0.400 68.638±23.978 41.9 YYS 8 13.771±5.095 86.766±15.225 6.3 YYS 9 15.113±4.749 94.029±19.713 6.2 YYS 10 44.763±11.696 113.069±15.837 2.5 YYS 11 27.460±8.477 >100 >3.6 YYS 12 34.431±10.804 >100 >2.9 YYS 13 NA 91.749±21.621 - YYS 14 56.342±15.866 >100 >1.8 YYS 15 31.550±9.281 68.192±21.097 2.2 YYS 16 60.243±17.001 50.418±10.361 0.8 YYS 17 NA >100 - YYS 18 35.429±12.115 78.724±24.168 2.2 YYS 19 2.519±0.763 >100 >48.3 YYS 20 1.338±0.318 >100 >93.0 YYS 21 NA 104.400±19.494 - YYS 22 25.148±8.928 81.429±16.879 3.2 Amantadine 0.551±0.161 >100 >181.5

The above experimental results show that most of the synthesized compounds have good anti-H5N1 activity, such as compounds YYS1 (EC ₅₀ =7.716±2.909μM, SI=14.9), YYS 4 (EC ₅₀ =5.302±1.450μM, SI>18.9) , YYS 5 (EC ₅₀ =9.970±2.548μM, SI=11.0), YYS 6 (EC ₅₀ =1.247±0.335μM, SI>80.2), YYS 7 (EC ₅₀ =1.639±0.400μM, SI=41.9), YYS 19 (EC ₅₀ = 2.519 ± 0.763 μM, SI>48.3), YYS 20 (EC ₅₀ = 1.338 ± 0.318 μM, SI = 93.0), etc., especially compounds YYS 6 and YYS 20 showed biological activities close to sub-micromolar levels .

Claims (4)

1. An aromatic formamide compound or a pharmacologically or physiologically acceptable salt thereof, wherein the compound is represented by the following compounds:

2. the aromatic carboxamide compound as claimed in claim 1, or a pharmacologically or physiologically acceptable salt thereof, which is characterized in that each of the compounds is represented by:

n- (2- ((4-chlorobenzyl) thio) ethyl) -2, 5-dimethylfuran-3-carboxamide,

N- (2- ((4-cyanobenzyl) thio) ethyl) -2, 5-dimethylfuran-3-carboxamide,

N- (2- ((3-nitrobenzyl) thio) ethyl) -2, 5-dimethylfuran-3-carboxamide,

N- (2- ((4-chlorobenzyl) thio) ethyl) -2, 5-dimethyl thiophene-3-formamide,

N- (2- ((4-fluorobenzyl) thio) ethyl) -2, 5-dimethylfuran-3-carboxamide,

N- (2- ((4-bromobenzyl) thio) ethyl) -2, 5-dimethylfuran-3-carboxamide,

2, 5-dimethyl-N- (2- ((4-nitrobenzyl) sulfenyl) ethyl) thiophene-3-formamide,

N- (2- ((4-cyanobenzyl) thio) ethyl) -2, 5-dimethylthiophene-3-carboxamide.

3. Use of the aromatic carboxamide compound as claimed in claim 1 or 2 or a pharmacologically or physiologically acceptable salt thereof for the preparation of a medicament against influenza a viruses.

4. A pharmaceutical composition against influenza a virus comprising the aromatic carboxamide compound as claimed in claim 1 or 2 or a pharmacologically or physiologically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.