Preparation and application of (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives

Abstract

The present invention discloses the preparation and application of (E)-N-[(1H-indazole-7-yl)methyl]-3-arylacrylamide derivatives, belonging to the field of medical technology. Specifically, the (E)-N-[(1H-indazole-7-yl)methyl]-3-arylacrylamide derivatives, their prodrugs, pharmaceutically active metabolites and pharmaceutically acceptable salts thereof are shown in the structural formula I, wherein Ar is as described in the claims and the specification. The corresponding (E)-N-[(1H-indazole-7-yl)methyl]-3-arylacrylamide derivatives and pharmaceutically applicable acids and pharmaceutically acceptable salts of such compounds can be combined with existing drugs or used alone as influenza virus inhibitors, and have good therapeutic effects on various influenza.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to an (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative and a preparation method thereof, as well as an application thereof in the preparation of an anti-influenza virus drug. Background Art

[0002] Influenza is an acute respiratory infectious disease caused by influenza virus, with the characteristics of high morbidity and high mortality (Loregian A, Mercorelli B, Nannetti G, et al. Antiviral strategies against influenza virus: toward new therapeutic approaches. Cellular and Molecular Life Sciences, 2014, 71 (19): 3659-3683.). At present, the main prevention and treatment measures for influenza virus are vaccination and the use of small molecule anti-influenza drugs. To adapt to antigenic variability, it is necessary to continuously develop new vaccines, but due to the relative lag in its development, and the inability to produce effective immunity to immunocompromised patients, small molecule drugs have become the main means of influenza prevention and treatment. Currently, there are three types of anti-influenza drugs approved by the US FDA: M2 ion channel blockers including amantadine and rimantadine; neuraminidase inhibitors including oseltamivir, zanamivir and peramivir; and baloxavirmarboxil, an acidic endonuclease inhibitor of influenza virus RNA-dependent RNA polymerase (RdRp), which was approved for marketing in October 2018. However, M2 ion channel resistance sites are widely found in influenza A virus, and alkylamine drugs are no longer used as clinical treatment drugs for influenza virus. The emergence of oseltamivir-resistant strains has also reduced its therapeutic effect; the bioavailability of peramivir is very low and it is only administered intravenously. For baloxavir mapositol, drug resistance has been reported in clinical trials and patients treated with the drug (Massari S, Goracci L, Desantis J, et al. Polymerase Acidic Protein-Basic Protein 1 (PA-PB1) Protein-Protein Interaction as a Target for Next-Generation Anti-influenza Therapeutics. Journal of Medicinal Chemistry, 2016, 59 (17): 7699-7718.). Therefore, it is urgent to develop new anti-influenza virus drugs.

[0003] RdRp is a heterotrimer composed of PA, PB1 and PB2. The PB1 subunit is located in the core of RNA polymerase. The N-terminus and C-terminus of the PB1 subunit bind to the C-terminus of the PA subunit and the N-terminus of the PB2 subunit, respectively, to form a stable protein complex. PB1 contains the RNA polymerase active site, PB2 is involved in the binding of the 5' cap structure of the host cell pre-mRNA, and PA has an endonuclease domain. The three subunits bind to each other and play a vital role in influenza virus RNA transcription and replication. The structure of RdRp is highly conserved among influenza A, influenza B, and influenza C virus strains, and there is no homologous protein in mammalian cells. Therefore, RNA polymerase has become an attractive target for anti-influenza drugs (He X, Zhou J, Bartlam M, et al. Crystal structure of the polymerase PA C -PB1 N complex from an avian influenza H5N1 virus. Nature, 2008, 454(7208): 1123-1126. Shen Z, LouK, Wang W. New small-molecule drug design strategies for fighting resistant influenza A. Acta Pharmaceutica Sinica B, 2015, 5(5): 419-430.).

[0004] Cinnamic acid and its derivatives have a wide range of biological activities, such as anticancer, antioxidant, antibacterial, anti-inflammatory, hypoglycemic, liver protection, antimalarial, antituberculosis, antifungal, antiviral, etc. Cinnamic acid is a class of active substances with Michael receptor fragments (Sova, M. Antioxidant antimicrobial activities of cinnamic acid derivatives. Mini-reviews in medicinal chemistry, 2012, 12 (8): 749-767.). Inspired by the structural fragments of cinnamic acid, the present invention selects aryl vinyl fragments with Michael receptor characteristics and designs a series of (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives. Through biological activity tests, it is expected that the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives have certain anti-influenza virus activity.

[0005] Therefore, the study of inhibitors targeting influenza virus RNA polymerase has potential advantages. The discovery of (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives as influenza virus inhibitors is of great significance for the expansion of the influenza virus inhibitor molecular library and the study of new influenza virus inhibitors. Summary of the invention

[0006] The technical problem solved by the present invention is to provide an (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative and its preparation and application, specifically the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative as shown in formula I, its prodrug and pharmaceutically active metabolite, and the stereoisomers of the compound and pharmaceutically acceptable salts thereof, and provide its application in the preparation of drugs for treating influenza-related diseases.

[0007] (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives having a structural formula as shown in Formula I, their prodrugs and pharmaceutically active metabolites, and stereoisomers of the compound and pharmaceutically acceptable salts thereof:

[0008]

[0009] Wherein, Ar is selected from halogen-substituted phenyl, halogen-substituted methylphenyl, nitro-substituted phenyl, methoxyphenyl, pyridyl, furyl, thienyl, the halogen includes fluorine, chlorine and bromine, and the number of the halogen atoms is 1-3.

[0010] Further, Ar is selected from 4-bromophenyl, 4-chlorophenyl, 4-trifluoromethylphenyl, 4-nitrophenyl, 4-fluorophenyl, 4-methoxyphenyl, 3,4-difluorophenyl, pyridin-4-yl, furan-2-yl, thiophen-2-yl.

[0011] Furthermore, the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative is any one of the following compounds:

[0012] (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-bromophenyl)acrylamide (JX01);

[0013] (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-chlorophenyl)acrylamide (JX02);

[0014] (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-trifluoromethylphenyl)acrylamide (JX03);

[0015] (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-nitrophenyl)acrylamide (JX04);

[0016] (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-fluorophenyl)acrylamide (JX05);

[0017] (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-methoxyphenyl)acrylamide (JX06);

[0018] (E)-N-[(1H-indazol-7-yl)methyl]-3-(3,4-difluorophenyl)acrylamide (JX07);

[0019] (E)-N-[(1H-indazol-7-yl)methyl]-3-(pyridin-4-yl)acrylamide (JX08);

[0020] (E)-N-[(1H-indazol-7-yl)methyl]-3-(furan-2-yl)acrylamide (JX09);

[0021] (E)-N-[(1H-indazol-7-yl)methyl]-3-(thiophen-2-yl)acrylamide (JX10).

[0022] The present invention also provides a pharmaceutical composition comprising a (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative as shown in formula I and a pharmaceutically acceptable carrier or diluent.

[0023] The pharmaceutical composition is used to prepare a drug for treating influenza virus diseases. For example, a saline aqueous solution is used as a carrier with a pH value of 7.4, and the pharmaceutical composition is introduced into the patient's bloodstream in the form of a solution by local injection.

[0024] The compounds of the present invention can be administered alone or in combination with pharmaceutically acceptable carriers or diluents according to conventional pharmaceutical practices in the form of a pharmaceutical composition. The administration routes include oral administration or parenteral administration including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical administration.

[0025] For oral administration, in addition to tablets or capsules, it can also be administered in the form of an aqueous solution or suspension. When administered orally, the active pharmaceutical ingredient is combined with an oral, non-toxic, pharmaceutically acceptable inert carrier to form a pharmaceutical composition. For oral administration in the form of tablets or capsules, the carrier includes lactose, starch, sucrose, glucose, methylcellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol; for oral administration in liquid form, the carrier includes ethanol, glycerol, water and a combination thereof. In addition, pharmaceutically acceptable binders, lubricants, disintegrants and colorants can also be added to the pharmaceutical composition. Binders include starch, gelatin, natural sugars such as glucose or lactose, corn sweeteners, natural and synthetic gums such as gum arabic, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, wax. Lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride. When an aqueous suspension is used orally, the active ingredient is combined with an emulsifier and a suspending agent. Sweeteners or flavoring agents may also be added. For intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared, and the pH of the solution should be appropriately adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled to maintain isotonicity of the preparation.

[0026] The (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives of the present invention can also be used in combination with known agents that can be used to treat or prevent influenza. Preferred combinations include compounds of the present invention and M2 ion channel protein inhibitors, compounds of the present invention and neuraminidase inhibitors, compounds of the present invention and interferon inducers, compounds of the present invention and antisense oligonucleotides, and compounds of the present invention and inosine monophosphate dehydrogenase inhibitors.

[0027] The term "administering" and its variants (e.g., "administering" a compound) in connection with the present invention means: introducing the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a prodrug of the compound into the system of an animal in need of treatment. When the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a prodrug of the compound is provided in combination with one or more other active agents, "administering" and its variants can be understood to include both simultaneous and sequential introduction of the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a prodrug of the compound and the other agent. Typically, the prodrug is a functional derivative of the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative that is readily converted into the desired compound in vivo. In the present invention, the term "administering" shall include administering the specifically disclosed (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives or compounds that may not be specifically disclosed but can be converted into specific compounds in vivo after administration to a patient.

[0028] When the compound of the present invention is administered to a human subject, the daily dose will generally be determined by the prescribing physician, and the dose generally varies according to the age, weight and reaction of the individual patient and the severity of the patient's symptoms. In an exemplary application, a suitable amount of the compound is administered to a mammal receiving treatment. When used for the indicated effect, the oral dose of the present invention will be about 0.01 mg per kg body weight per day (mg / kg / day) to about 100 mg / kg / day, preferably 0.01 mg / kg / day to 10 mg / kg / day, and most preferably 0.1 mg / kg / day to 5.0 mg / kg / day. For oral administration, the composition is preferably provided in the form of a tablet, wherein the tablet comprises 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100 and 500 mg of active ingredient, preferably 1 mg to 100 mg of active ingredient. For intravenous injection, during constant rate infusion, the most preferred dosage is 0.1 mg / kg / min to 10 mg / kg / min. The compound of the present invention or the pharmaceutical composition comprising the compound can be administered in a dose once a day, or the total daily dose can be divided into two, three or four doses per day. For administration in the form of a transdermal delivery system, the dosage administration will certainly be continuous rather than intermittent throughout the dosage regimen.

[0029] The dosage regimen using the compounds of the present invention will be selected based on a variety of factors, including the type, species, age, weight, sex and medical condition of the patient; the severity of the condition being treated; the route of administration; the patient's renal and liver function; and the specific compound or salt thereof used. An ordinarily skilled physician, veterinarian or clinician can readily determine and prescribe the effective amount of the drug required to prevent, combat or arrest the development of a condition.

[0030] In the method of the present invention, the compounds described in detail herein can form the active ingredient and be mixed with a suitable pharmaceutical diluent, excipient or carrier (collectively referred to herein as "carrier" material) appropriately selected according to the form of administration, i.e., oral tablets, capsules, elixirs, syrups, etc., and in accordance with conventional pharmaceutical practices.

[0031] The pharmaceutically acceptable salts of the compounds of the present invention refer to conventional acid addition salts or base addition salts which retain the biological effectiveness and properties of the compounds of formula I and are formed with suitable non-toxic organic or inorganic acids or organic or inorganic bases. Acid addition salts include hydrochlorides, hydrobromides, hydroiodides, nitrates, phosphates, sulfates, perchlorates, thiocyanates, bisulfates, persulfates, borates, formates, acetates, propionates, valerates, pivalates, caproates, heptanoates, caprylates, isooctanoates, undecanoates, laurates, palmitates, stearates, oleates, cyclopropionates, oxalates, malonates, succinates, maleates, fumarates, adipates, azelaates, acrylates, crotonates, tiglates, itaconates, sorbates, cinnamates, glycolates, lactates, malates, tartrates, citrates, tartrates, mandelates, benzilates, tropicates, ascorbic acids, gluconates, glucoheptonates, glucarates, mannonates, lactobionates, benzoates, phthalates, terephthalates, furoates, nicotinates, isonicotinates, Salicylate, acetylsalicylate, butyrate, gallate, caffeate, ferulate, picrate, camphorate, camphorsulfonate, methanesulfonate, ethanesulfonate, propanesulfonate, benzenesulfonate, p-toluenesulfonate, p-aminobenzenesulfonate, sulfamate, taurine, 2-hydroxyethanesulfonate, glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartate, asparagine, glutamate, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine, arginine, edetate, pyruvate, α-ketoglutarate, alginate, cyclopentanepropionate, 3-phenylpropionate, 3-cyclohexylpropionic acid, 2-naphthoate, 2-naphthalenesulfonate, pamoate, lauryl sulfate, glycerophosphate, lauryl sulfate, pectinate, etc. Base addition salts include ammonium salts, alkali metal salts, such as sodium and potassium salts, alkaline earth metal salts, such as calcium and magnesium salts, salts with organic bases, such as dicyclohexylamine salts, N-methyl-D-glucamine salts, and the like. Moreover, basic nitrogen-containing groups can be quaternized with such agents as lower alkyl halides, such as the chloride, bromide and iodide of methyl, ethyl, propyl and butyl; dialkyl sulfates, such as dimethyl sulfate, diethyl, dibutyl and diamyl sulfate; long chain halides, such as the chloride, bromide and iodide of decyl, lauryl, myristyl and stearyl; aralkyl halides, such as the bromide of benzyl and phenethyl, and the like.

[0032] The present invention also provides a method for preparing the (E)-N-[(1H-indazol-7-yl)methyl]-3-aryl acrylamide derivative shown in general formula I, comprising the following steps:

[0033]

[0034] Beneficial effects of the present invention

[0035] The compound provided by the present invention is used to inhibit influenza virus and is a new type of influenza virus inhibitor acting on RNA polymerase. Compared with the anti-influenza virus drugs currently on the market that act on other targets, the influenza virus inhibitor acting on RNA polymerase has stability. This type of influenza virus inhibitor compound has a wide range of therapeutic effects on influenza. In addition, the preparation method of the compound is simple, the yield is stable, and the prepared compound can better treat influenza-related diseases. DETAILED DESCRIPTION

[0036] The technical solution of the present invention is described in detail with the following embodiments. However, it should be understood that the present invention is not limited to the following examples specifically described.

[0037] Example 1: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-bromophenyl)acrylamide (JX01)

[0038]

[0039] 0.68g (3.00mmol) of 3-(4-bromophenyl)acrylic acid, 1.37g (3.60mmol) of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), and 30mL of N,N-dimethylformamide (DMF) were added to a 50mL eggplant-shaped bottle. After reacting at room temperature for 1 hour, 0.53g (3.60mmol) of 1H-indazole-7-methylamine and 0.58g (4.50mmol) of N,N-diisopropylethylamine (DIPEA) were added. The reaction was continued at room temperature. TLC monitored the complete reaction of the raw materials (24 hours). The reaction solution was added to 100mL of water, extracted with 20mL of ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. Column chromatography (dichloromethane: methanol = 150: 1 v / v) gave 0.25g of a white solid with a yield of 88.0%. mp:214.5-215.4℃; 1 H-NMR (400MHz, DMSO-d6); δ13.05(s,1H),8.75(t,J=5.9Hz,1H),8.11(s,1H),7.69-7.67(m,1H),7.63-7.61(m,2H ),7.57-7.45(m,3H),7.27-7.19(m,1H),7.09-7.07(m,1H),6.77(d,J=15.8Hz,1H),4.71(d,J=5.9Hz,2H); ESI-MS m / z:356.0([M+H] + ).

[0040] Example 2: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-chlorophenyl)acrylamide (JX02)

[0041]

[0042] Referring to the preparation method of Example 1, 0.22 g of white solid was obtained with a yield of 87.6%. mp: 197.7-198.8°C; 1 H-NMR (400MHz, DMSO-d6); 13.05 (s, 1H), 8.75 (t, J = 5.9Hz, 1H), 8.12 (s, 1H), 7.69-7.67 (m, 1H), 7.63-7.61 (m, 2H ),7.56-7.44(m,3H),7.25-7.23(m,1H),7.13-7.06(m,1H),6.75(d,J=15.8Hz,1H),4.72(d,J=5.9Hz,2H); ESI-MS m / z:312.1([M+H] + ).

[0043] Example 3: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-trifluoromethylphenyl)acrylamide (JX03)

[0044]

[0045] Referring to the preparation method of Example 1, 0.16 g of white solid was obtained with a yield of 88.2%. mp: 213.8-214.8°C; 1 H-NMR (400MHz, DMSO-d6); δ13.07(s,1H),8.82(t,J=5.7Hz,1H),8.12(s,1H),7.85-7.74(m,4H),7.70-7.68(m,1H), 7.59(d,J=15.8Hz,1H),7.30-7.21(m,1H),7.15-7.04(m,1H),6.88(d,J=15.8Hz,1H),4.73(d,J=5.7Hz,2H); ESI-MS m / z:346.1([M+H] + ).

[0046] Example 4: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-nitrophenyl)acrylamide (JX04)

[0047]

[0048] Referring to the preparation method of Example 1, 0.17 g of white solid was obtained with a yield of 88.1%. mp: 262.8-263.4°C; 1 H-NMR (400MHz, DMSO-d6); δ13.07(s,1H),8.87(t,J=5.7Hz,1H),8.35-8.19(m,2H),8.12(s,1H),7.92-7.80(m,2H ),7.73-7.55(m,2H),7.31-7.20(m,1H),7.14-7.06(m,1H),6.94(d,J=15.9Hz,1H),4.74(d,J=5.7Hz,2H); ESI-MS m / z:323.1([M+H] + ).

[0049] Example 5: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-fluorophenyl)acrylamide (JX05)

[0050]

[0051] Referring to the preparation method of Example 1, 0.19 g of white solid was obtained with a yield of 89.4%. mp: 175.5-176.6°C; 1 H-NMR (400MHz, DMSO-d6); δ13.04(s,1H),8.72(t,J=5.9Hz,1H),8.12(s,1H),7.72-7.60(m,3H),7.52(d,J=15 .8Hz,1H),7.33-7.19(m,3H),7.14-7.04(m,7.0Hz,1H),6.69(d,J=15.8Hz,1H),4.71(d,J=5.9Hz,2H); ESI-MS m / z:296.1([M+H] + ).

[0052] Example 6: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-methoxyphenyl)acrylamide (JX06)

[0053]

[0054] Referring to the preparation method of Example 1, 0.18 g of white solid was obtained with a yield of 87.7%. mp: 168.7-169.7°C; 1H-NMR (400MHz, DMSO-d6); δ13.03(s,1H),8.65(t,J=5.8Hz,1H),8.11(s,1H),7.68-7.66(m,1H),7.57-7.51(m,2H),7.47(d,J=15.8 Hz,1H),7.6-7.20(m,1H),7.13-7.05(m,1H),7.01-6.95(m,2H),6.60(d,J=15.8Hz,1H),4.70(d,J=5.8Hz,2H),3.79(s,3H); ESI-MS m / z:308.1([M+H] + ).

[0055] Example 7: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(3,4-difluorophenyl)acrylamide (JX07)

[0056]

[0057] Referring to the preparation method of Example 1, 0.15 g of white solid was obtained with a yield of 82.3%. mp: 210.5-211.2°C; 1 H-NMR (400MHz, DMSO-d6); δ13.05(s,1H),8.72(t,J=5.6Hz,1H),8.12(d,J=1.4Hz,1H),8.11(s,1H),7.69-7.66(m,2H),7. 62-7.61(m,1H),7.41-7.40(m,1H),7.26-7.22(m,1H),7.10(m,2H),6.50(d,J=15.5Hz,1H),4.70(d,J=5.7Hz,2H); ESI-MS m / z:314.1([M+H] + ).

[0058] Example 8: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(pyridin-4-yl)acrylamide (JX08)

[0059]

[0060] Referring to the preparation method of Example 1, 0.17 g of white solid was obtained with a yield of 88.2%. mp: 189.5-190.1°C; 1H-NMR (400MHz, DMSO-d6); δ13.05(s,1H),8.74(t,J=5.8Hz,1H),8.12(s,1H),7.76-7.65(m ,2H),7.55-7.43(m,3H),7.09-7.08(m,1H),6.73(d,J=15.8Hz,1H),4.73(d,J=5.8Hz,2H). 13 C NMR(100MHz,DMSO-d6)δC:165.51,150.67,149.63,138.95,136.39,134.47,13 4.36,131.09,124.54,124.44,124.30,123.47,121.81,120.84,119.81; ESI-MS m / z:279.1([M+H] + ).

[0061] Example 9: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(furan-2-yl)acrylamide (JX09)

[0062]

[0063] Referring to the preparation method of Example 1, 0.19 g of white solid was obtained with a yield of 88.1%. mp: 175.8-176.6°C; 1 H-NMR (400MHz, DMSO-d6); δ13.04(s,1H),8.77(t,J=5.8Hz,1H),8.11(s,1H),7.79-7.78(m,1H),7.68- 7.67(m,1H),7.35-7.31(m,1H),7.23-7.21(m,1H),7.13-7.05(m,1H),6.81-6.80(m,J=3.3Hz,1H),6.60 -6.59(m,J=3.3,1H),4.70(d,J=5.8Hz,2H); ESI-MSm / z:267.2([M+H] + ).

[0064] Example 10: Preparation of (E)-N-[(1H-indazol-7-yl)methyl]-3-(thiophen-2-yl)acrylamide (JX10)

[0065]

[0066] Referring to the preparation method of Example 1, 0.15 g of white solid was obtained with a yield of 82.3%. mp: 186.2-186.9°C; 1H-NMR (400MHz, DMSO-d6); δ13.04(s,1H),8.72(s,1H),8.11(s,1H),7.69-7.66(m,2H),7.62-7.61(m,1H),7. 41-7.40(m,1H),7.26-7.24(m,1H),7.16-7.05(m,2H),6.50(d,J=15.5Hz,1H),4.70(d,J=5.8Hz,2H); ESI-MS m / z:284.1([M+H] + ).

[0067] Example 11: Cytotoxicity of target compounds

[0068] The target compounds JX01-JX10 were tested for cytotoxicity using the MTT method based on Madin-Daby canine kidney cells (MDCK), and the cytotoxicity of the target compounds was investigated at the cellular level. After adding different concentrations of the target compounds to the MDCK cell culture medium, the absorbance OD value was read by an enzyme-linked immunosorbent assay, and the survival rate of the MDCK cells was calculated (MDCK cell survival rate = absorbance value of the experimental group / absorbance value of the blank control group). The results are shown in Table 1. The lower the MDCK cell survival rate, the stronger the inhibitory effect of the target compound on the growth of MDCK cells, and the greater the cytotoxicity of the compound.

[0069] Table 1 Inhibitory effects of target compounds on MDCK cells

[0070]

[0071]

[0072] The pharmacological data showed that JX01 and JX10 significantly inhibited the survival of MDCK cells. -1 , 50μmol·L -1 , 25μmol·L -1 , 12.5 μmol·L -1 The survival rates of MDCK cells were 48.63%, 49.17%, 52.93% and 57.10% respectively. -1 , 50μmol·L -1 , 25μmol·L -1 , 12.5 μmol·L -1Under the conditions of 100 μg / mL, the cell survival rates were 39.43%, 53.17%, 44.65% and 64.23% respectively. The inhibitory effects of the other compounds on MDCK cells were not obvious.

[0073] Example 12: Protective effect of compounds on diseased cells

[0074] The MTT method was used to investigate the protective effect of the target compound on the diseased cells infected by influenza virus. After adding different concentrations of the target compound to the infected cell culture medium, the absorbance OD value was read by an enzyme-linked immunosorbent assay, and the survival rate of the diseased cells was calculated (pathological cell survival rate = absorbance value of the experimental group / absorbance value of the virus-free control group). The results are shown in Table 2. The higher the survival rate of the diseased cells, the stronger the protective ability of the target compound on the diseased cells and the stronger the activity of inhibiting influenza virus.

[0075] Table 2 Protective ability of target compounds against diseased cells

[0076]

[0077] The experimental results showed that compounds JX08 and JX10 had a certain protective ability against diseased cells, while the other compounds had a weaker protective ability against diseased cells. -1 , 25μmol·L -1 , 50μmol·L -1 , 100 μmol·L -1 At the concentration of 12.5 μmol·L -1 , 25μmol·L -1 , 50μmol·L -1 , 100 μmol·L -1 Under the different treatment modes, the survival rates of diseased cells were 31.73%, 32.77%, 38.83% and 41.40% respectively.

[0078] Through the cytotoxicity test and pathological cell protection test results of the above target compounds, it can be seen that:

[0079] (1) By comparing the structures of JX01, JX02, and JX05 with those of the other compounds, we found that cytotoxicity increased when the terminal benzene ring was substituted with bromine, while cytotoxicity decreased when the substituent was chlorine or fluorine.

[0080] (2) When the terminal benzene ring is replaced by a bioisosteric pyridine ring, furan ring, or thiophene ring, compounds JX08, JX09, and JX10 are obtained. When the terminal is substituted with a furan ring, compound JX09 has a certain protective ability against diseased cells, and its toxicity to normal cells is also low; when the terminal is substituted with a thiophene ring, compound JX10 has relatively good anti-influenza virus activity, but its cytotoxicity is also increased. In general, when the terminal is substituted with an aromatic heterocyclic ring, the activity is slightly better than that of the benzene ring.

[0081] Example 13: Compound Inhibition of Influenza Virus Experiment

[0082] The inhibitory activity of the target compound on influenza virus was tested by standard plaque assay. MDCK cells were inoculated into 96-well cell culture plates at a certain concentration, and the cells were cultured at 5% CO2 and 37°C for 24 hours. After that, the culture medium was removed, and the influenza virus was inoculated into the MDCK cells and adsorbed at 37°C for 2 hours. The virus solution was poured off, and the target compound was added. Three replicates were made for each concentration. A virus control group (infected with virus, without adding the target compound) was set up and continued to be cultured at 37°C for 24 hours, and the number of virus strains in the cells was determined by the standard plaque assay of MDCK cells. The results of the inhibition of the target compound on influenza virus strains at a concentration of 50 μmol / L are shown in Table 3.

[0083] Table 3 Effects of compounds at a concentration of 50 μmol / L on the survival rates of three influenza virus strains

[0084]

[0085] The target compounds were initially screened for anti-influenza virus activity through influenza virus inhibition experiments. At a concentration of 50 μmol / L, the target compounds had varying degrees of inhibitory effects on influenza virus strains A / WSN / 33 (H1N1), A / PR / 8 / 34 (H1N1) and A / HK / 68 (H3N2). Among them, compounds JX08, JX09 and JX10 performed best, with a survival rate of influenza virus strains below 70%.

[0086] In the following formulations, "active ingredient" refers to the compound represented by Formula I, or a salt or solvate thereof.

[0087] Example 14: Gelatin Capsules

[0088]

[0089] Example 15: Tablets

[0090]

[0091] Example 16: Tablet

[0092]

[0093] The active ingredient, starch and cellulose are passed through a 45 mesh US sieve and mixed thoroughly. The resulting powder is mixed with polyvinylpyrrolidone and then passed through a 14 mesh US sieve. The granules thus obtained are dried at 50-60°C and passed through a 18 mesh US sieve. Sodium carboxymethylcellulose, magnesium stearate and talc are first passed through a 60 mesh US sieve and then added to the above granules. After mixing, they are compressed into tablets on a tablet press.

[0094] Example 17: Suspension

[0095]

[0096] The drug is passed through a No. 45 mesh US sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a uniform paste. The benzoic acid solution, flavoring agent, and coloring agent are diluted with some of the water and added with stirring. Sufficient water is then added to produce the required volume.

[0097] Example 18: Aerosol

[0098]

[0099] The active ingredient is mixed with ethanol and the resulting mixture is added to the propellant 22, cooled to 30°C and transferred to a container. The required amount is then added to a stainless steel container and diluted with the remaining propellant, and then the valve assembly is installed.

[0100] Example 19: Suppository

[0101]

[0102] The active ingredient is passed through a No. 60 mesh US sieve and suspended in the pre-melted saturated fatty acid glycerides. The mixture is then poured into standard 2 g cavity suppository molds and allowed to cool.

[0103] Example 20: Injectable Formulation

[0104]

[0105] The above solution was administered to the patient by intravenous injection at a rate of about 1 mL / min.

[0106] The above is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still belongs to the protection scope of the technical solution of the present invention.

Claims

1. A (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a pharmaceutically acceptable salt thereof having a structural formula as shown in Formula I: in, Ar is selected from halogen-substituted phenyl, halogen-substituted methylphenyl, nitro-substituted phenyl, methoxyphenyl, pyridyl, furyl, thienyl, the halogen includes fluorine, chlorine and bromine, and the number of the halogen atoms is 1-3.

2. The (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: Ar is selected from 4-bromophenyl, 4-chlorophenyl, 4-trifluoromethylphenyl, 4-nitrophenyl, 4-fluorophenyl, 4-methoxyphenyl, 3,4-difluorophenyl, pyridin-4-yl, furan-2-yl, thiophen-2-yl.

3. The (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: The compound is selected from one of the following compounds: (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-bromophenyl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-chlorophenyl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-trifluoromethylphenyl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-nitrophenyl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-fluorophenyl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(4-methoxyphenyl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(3,4-difluorophenyl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(pyridin-4-yl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(furan-2-yl)acrylamide; (E)-N-[(1H-indazol-7-yl)methyl]-3-(thiophen-2-yl)acrylamide.

4. The method for preparing the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The following steps are involved:

5. A pharmaceutical composition, characterized in that A compound comprising as an active ingredient any one of the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivatives according to any one of claims 1 to 3 or any one of the pharmaceutically acceptable salts of the compound and a pharmaceutically acceptable carrier or diluent.

6. Use of the (E)-N-[(1H-indazol-7-yl)methyl]-3-arylacrylamide derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a drug for treating influenza virus.

7. Use of the pharmaceutical composition according to claim 5 in preparing a drug for treating influenza virus.

8. The use according to claim 6, characterized in that: An effective amount of the compound acts on RNA polymerase of influenza virus to inhibit the replication of influenza virus.

9. The use according to claim 7, characterized in that: An effective amount of the pharmaceutical composition acts on the RNA polymerase of the influenza virus to inhibit the replication of the influenza virus.

Images