Capsid inhibitor and its preparation method and use

Abstract

The present invention discloses a capsid inhibitor and a preparation method and use thereof. The capsid inhibitor of the present invention can occupy the central hole of the viral capsid protein hexamer, thereby inhibiting the introduction of nucleotides, thereby effectively blocking the reverse transcription in the viral capsid, and at the same time interfering with the formation of the capsid protein lattice, and has excellent antiviral activity, for example, showing significant anti-HIV activity.

Description

Technical Field

[0001] The present invention relates to the field of antiviral drugs, and in particular to a capsid inhibitor and a preparation method and use thereof. Background Art

[0002] AIDS, also known as "Acquired immunodeficiency syndrome" (AIDS), is a malignant disease caused by human immunodeficiency virus (HIV) infecting T cells, resulting in defects in the body's defense function, which in turn leads to various opportunistic infections and even tumors. At present, the clinical anti-AIDS therapeutic drugs mainly include the following eight categories: adsorption inhibitors, CCR5 antagonists, non-nucleoside reverse transcriptase inhibitors, nucleoside reverse transcriptase inhibitors, integrase chain transfer inhibitors, protease inhibitors, capsid regulators and pharmacokinetic enhancers. In order to reduce the impact of viral resistance and improve the efficacy of treatment, three or more drugs that have effects on different links of HIV replication are usually used in combination to treat AIDS, namely "highly effective antiretroviral therapy". Although this therapy can maximally inhibit viral replication and reduce the HIV load in patients, inhibitors have problems such as drug resistance after long-term use, poor patient compliance, and serious toxic side effects. Therefore, developing new, highly effective, and low-toxic anti-HIV drugs with new mechanisms or binding to new sites for targets with druggable potential is an effective way to overcome the problem of drug resistance in existing therapies and enrich clinical treatment options.

[0003] HIV capsid protein is a structural protein necessary for the formation of a morphologically mature and infectious virus particle. It contains RNA genome and multiple key enzymes that are essential for viral replication. In addition, it is a multifunctional protein that plays an important regulatory role in multiple links such as reverse transcription, nuclear entry, uncoating in the early stage of viral replication, and assembly and maturation in the late stage. Further studies have found that capsid protein can form a hexamer with a central pore, which can recruit nucleotides into viral particles in the early stage, thereby promoting the synthesis of DNA in the capsid. In the late stage, it interacts with IP6 to assemble the capsid into mature viral particles. Capsid protein has become a highly anticipated target for anti-HIV drug research, but there is currently no clinical drug targeting the central pore of the capsid protein hexamer.

[0004] The information in the background technology is only for illustrating the general background of the present invention and should not be regarded as admitting or suggesting in any form that such information constitutes the prior art known to a person skilled in the art. Summary of the invention

[0005] In order to solve at least some of the technical problems in the prior art, the present invention has conducted in-depth research and designed and developed a new compound for the central hole of the capsid protein hexamer. The new compound obtained by the present invention is a small molecule compound that can occupy the central hole site of the capsid protein hexamer, thereby inhibiting nucleotide introduction, thereby effectively blocking reverse transcription in the viral capsid, and interfering with the formation of the capsid protein lattice. Specifically, the present invention includes the following contents.

[0006] In a first aspect of the present invention, a capsid inhibitor is provided, which has a structure shown in the following formula (I):

[0007] Formula (I);

[0008] Wherein: R1 represents a carboxyl group or an ester group, and R2 represents a substituted phenyl group; wherein the substituted phenyl group includes a phenyl group in which at least one hydrogen on the benzene ring is replaced by a hydroxyl group.

[0009] In certain embodiments, according to the capsid inhibitor of the present invention, the substituted phenyl group is selected from one of the following:

[0010] .

[0011] In certain embodiments, the capsid inhibitor according to the present invention has a structural formula preferably selected from one of the following:

[0012] , , , , , .

[0013] The second aspect of the present invention provides a method for preparing a capsid inhibitor, which comprises the following steps:

[0014] (1) reacting a thiazolidinedione with a halogen-substituted alkyl ester nucleophile to obtain an intermediate; and

[0015] (2) further reacting the intermediate with a substituted benzaldehyde to generate an inhibitor compound, wherein at least one hydrogen on the benzene ring of the substituted benzaldehyde is replaced by a hydroxyl group.

[0016] In certain embodiments, the method for preparing the capsid inhibitor according to the present invention further comprises the step of subjecting the inhibitor compound obtained in step (2) to ester hydrolysis reaction.

[0017] In certain embodiments, according to the method for preparing a capsid inhibitor of the present invention, the alkyl ester comprises a C1-C10 alkyl acid.

[0018] The third aspect of the present invention provides a pharmaceutical composition comprising the capsid inhibitor described in the first aspect and a pharmaceutically acceptable carrier.

[0019] The fourth aspect of the present invention provides an in vitro method for inhibiting or resisting viruses, which comprises the step of contacting the capsid inhibitor described in the first aspect with viruses in vitro.

[0020] The fifth aspect of the present invention provides use of the capsid inhibitor described in the first aspect in the preparation of an antiviral drug, wherein the virus comprises a capsid protein, in particular a capsid protein hexamer.

[0021] The capsid inhibitors of the present invention have excellent antiviral activity, for example, exhibit significant anti-HIV activity, among which compounds ZA-29 and ZA-30 have particularly outstanding activity. DETAILED DESCRIPTION

[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0023] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that the upper and lower limits of the scope and each intermediate value therebetween are specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0024] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which the invention belongs. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0025] [Capsid inhibitors]

[0026] In the first aspect of the present invention, a capsid inhibitor is provided, sometimes referred to herein as "the inhibitor of the present invention", which is a compound that can act on viral capsid protein, especially capsid protein derived from human immunodeficiency virus, and regulate its activity, especially regulate its multimer activity such as hexamer. Exemplarily, the regulation of hexamer activity of the present invention includes but is not limited to occupying the central pore site of the capsid protein hexamer, thereby inhibiting nucleotide introduction, thereby effectively blocking the reverse transcription of viruses such as HIV-1 in the capsid, and also includes interfering with the formation of capsid protein lattice.

[0027] The inhibitor of the present invention is a compound having a structure shown in the following formula (I):

[0028] Formula (I);

[0029] Wherein: R1 represents a carboxyl group or an ester group, and R2 represents a substituted phenyl group.

[0030] As used herein, the term "carboxy" refers to a carboxylic acid-containing group, such as -COOH.

[0031] Herein, the term "ester group" refers to a structure obtained by the reaction of an alcohol and a carboxylic acid group, examples of which include esters formed by a carboxyl group and a C1-C20 short-chain alcohol, such as a short-chain monohydric alcohol. Short-chain alcohols include short straight-chain or short-branched alcohols. Exemplary ester group examples include carboxymethyl ester, carboxyethyl ester, carboxy-n-propyl ester, carboxy-isopropyl ester, carboxy-n-butyl ester, carboxy-isobutyl ester, carboxy-n-pentyl ester, carboxy-isopentyl ester, etc.

[0032] Herein, the term "substituted phenyl" refers to a phenyl structure in which at least one hydrogen on a phenyl ring is substituted, and the substituent replacing hydrogen includes a hydroxyl group. In certain embodiments, the substituted phenyl group includes a hydroxyl group, and its position on the phenyl ring is not limited, and can be any position. In certain embodiments, the substituted phenyl group includes two or more hydroxyls. At this time, the position of two or more hydroxyls is not limited, and can be any position of the phenyl ring. Exemplarily, the present invention is two hydroxyls, and the two hydroxyls are adjacent positions, or spaced positions. In certain embodiments, the phenyl ring of the present invention each contains a hydroxyl group in adjacent positions, and the two hydroxyls form a ring structure.

[0033] In certain embodiments, the substituted phenyl groups of the present invention are selected from one of the following:

[0034] .

[0035] In certain preferred embodiments, the capsid inhibitor of the present invention has a structural formula selected from one of the following:

[0036] , , , , , .

[0037] [Method for preparing capsid inhibitor]

[0038] The second aspect of the present invention provides a method for preparing a capsid inhibitor, which is mainly prepared by using a thiazolidinedione compound as a raw material through a nucleophilic reaction and a reaction with a substituted benzaldehyde. The method of the present invention comprises at least the following steps:

[0039] (1) reacting thiazolidinedione with a halogen-substituted alkyl ester nucleophile to obtain an intermediate;

[0040] (2) allowing the intermediate to further react with a substituted benzaldehyde to generate a compound;

[0041] Optionally, the method further comprises the step of (3) subjecting the compound obtained in step (2) to ester hydrolysis reaction. Each step is described in detail below.

[0042] Step (1):

[0043] The step (1) of the present invention is to react thiazolidinedione as a raw material with a nucleophilic reagent to obtain an intermediate. Wherein, the nucleophilic reagent is generally a halogen-substituted alkyl ester. The halogen as a substituent group is not limited, and includes but is not limited to fluorine, chlorine, bromine, iodine, etc. The number of halogen substitutions is not limited, and preferably there is only one halogen. The halogen substitution position is not limited. In the case of only one halogen substitution, it is usually located at one end of the alkyl ester.

[0044] Herein, the term "alkyl ester" refers to a compound obtained by reacting a short-chain alkyl acid with an alcohol (preferably a monohydric alcohol). The alcohol herein is not limited, and includes, for example, C1-C10 alcohols, preferably C1-C6 straight-chain or branched alcohols, examples of which include, but are not limited to, methanol, ethanol, propanol, butanol, pentanol, etc. The alkyl acid herein is not limited, and includes C1-C20 alkyl acids, such as formic acid, acetic acid, propionic acid, butyric acid, valeric acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, etc.

[0045] The step (1) of the present invention is usually carried out in a solvent when reacting, and the solvent is generally a solvent that can dissolve the thiazolidinedione and the nucleophilic reagent at the same time. Such solvents are generally lipophilic solvents, such as amide solvents, examples of which include but are not limited to dimethylformamide, dimethylacetamide, etc. When performing step (1), a soluble carbonate is usually further added to the solvent, examples of which include but are not limited to potassium carbonate, sodium carbonate, magnesium carbonate, etc. The present invention can use one or more of the above-mentioned carbonates. The amount of carbonate used is not limited, and those skilled in the art can freely add it as needed.

[0046] The reaction temperature of step (1) of the present invention is not limited, and generally there is no need to specifically control the temperature, which is usually in the range of 5-30°C, such as 6°C, 10°C, 12°C, 14°C, 15°C, 16°C, 18°C, 20°C, 22°C, 24°C, 25°C, 26°C, 28°C, etc. Exemplarily, step (1) of the present invention is carried out at room temperature. The reaction time of step (1) of the present invention is not limited, and is usually 1-30 hours, such as 5-25 hours, 8-20 hours, 10-18 hours, 12-15 hours, etc. It should be noted that the above reaction time is only an exemplary time. In some cases, the reaction time may also be less than 1 hour, or greater than 30 hours.

[0047] Step (2):

[0048] The step (2) of the present invention includes the step of further reacting the intermediate with a substituted benzaldehyde to generate a compound. The step (2) is preferably carried out in a solvent, and the solvent of the step (2) includes a polar solvent, such as an alcohol solvent, a ketone solvent, an ether solvent, and the like.

[0049] The step (2) of the present invention is preferably carried out in the presence of a catalyst. The catalyst is not limited and any catalyst known in the art may be used, examples of which include but are not limited to piperidine.

[0050] The step (2) of the present invention is preferably carried out under reflux conditions. The reflux reaction time is not limited, and is generally 1-20 hours, for example, 2-18 hours, 3-15 hours, 5-12 hours, such as 6 hours, 8 hours, 10 hours, etc. It should be noted that the above reaction time is only an exemplary time, and in some cases, the reaction time may also be less than 1 hour, or greater than 20 hours. Those skilled in the art can freely select the reaction time according to the specific circumstances.

[0051] Step (3):

[0052] Step (3) of the present invention is a step of hydrolyzing to obtain an acid, which is an optional step. Specifically, step (3) includes hydrolyzing the compound obtained in step (2) under acidic conditions. The "acidic conditions" herein include reacting the product of step (2) in an acid solution. "Acid solution" herein refers to an acid solution or a mixed solution thereof with an aqueous solvent such as water. The type of acid is not limited, and examples thereof include organic acids such as formic acid, acetic acid, propionic acid, and inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, etc. The present invention can use one of the above acids, or a combination of multiple thereof. In certain embodiments, the acid of the present invention is a mixture of acetic acid and hydrochloric acid, and preferably the volume ratio of the two is in the range of 1:1 to 10:1. The amount ratio of the acid to the aqueous solvent in the acid solution is not limited, and those skilled in the art can freely choose according to needs.

[0053] It should be understood by those skilled in the art that, herein, the numbering of steps (1), (2) or (3) is only for the purpose of distinguishing different steps, and does not mean the order of the steps. As long as the purpose of the present invention can be achieved, the order of the above steps is not particularly limited. In addition, it should be understood by those skilled in the art that other steps or operations may be included before or after the above steps (1)-(3), or between these arbitrary steps, for example, to further optimize and / or improve the method described in the present invention.

[0054] In certain embodiments, the preparation method of the present invention comprises: using DMF as a solvent, 2,4-thiazolidinedione and 6-bromohexanoic acid methyl ester undergo a nucleophilic reaction to generate an intermediate Z2; then, using ethanol as a solvent, reacting with a substituted benzaldehyde under the catalysis of piperidine to generate ZA-26, ZA-28 and ZA-30; finally, undergoing an ester hydrolysis reaction with acetic acid / hydrochloric acid in water to obtain ZA-25, ZA-27 and ZA-29.

[0055]

[0056] Reagents and conditions: (i) potassium carbonate, DMF, room temperature, 12 h; (ii) piperidine, ethanol, reflux, 8 h; (iii) acetic acid:hydrochloric acid (3:1), water, 100°C, 16 h.

[0057] In an exemplary embodiment, the method of the present invention is the synthesis of the target compound ZA-25-30, which comprises:

[0058] (1) Add methyl 6-bromohexanoate to 50 mL of DMF, then add potassium carbonate and 2,4-thiazolidinedione, and react at room temperature under magnetic stirring. After 12 h, the reaction is complete as monitored by TLC, and the solvent DMF is removed by rotary evaporation. Ethyl acetate is added to the reaction bottle and the reaction system is transferred to a 250 mL separatory funnel, and water is added for extraction, and the lower organic phase is collected. The aqueous phase is washed with ethyl acetate, and the organic phases are combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove ethyl acetate. The crude product is mixed with 100-200 mesh silica gel and separated by column chromatography to obtain a white oil Z2.

[0059] (2) The intermediate Z1 was added to 10 mL of ethanol, and then substituted benzaldehyde and 0.5 mL of piperidine were added, and the mixture was refluxed at 80°C under magnetic stirring. After 8 h, the reaction was completed by TLC monitoring. 1 M hydrochloric acid was added to the reaction flask to adjust the pH to neutral. Solids precipitated and were filtered to obtain ZA-26, ZA-28 and ZA-30.

[0060] (3) ZA-26, ZA-28 and ZA-30 were added to 3 mL of water and placed under magnetic stirring. Then 3 mL of acetic acid and 1 mL of water were added to the bottle and the temperature was raised to 100 °C for reflux reaction. After 12 h, the reaction was completed by TLC monitoring. The solvent was removed by rotary evaporation, and then 5 mL of water was added and stirred at room temperature for 10 min. The mixture was filtered to obtain ZA-25, ZA-27 and ZA-29.

[0061] [Drug composition]

[0062] The third aspect of the present invention provides a pharmaceutical composition comprising the capsid inhibitor described in the first aspect of the present invention and a pharmaceutically acceptable carrier.

[0063] In the present invention, pharmaceutically acceptable carriers are well known in the art, and those skilled in the art can determine whether they meet clinical standards. Pharmaceutically acceptable carriers include diluents and excipients.

[0064] In certain embodiments, examples of suitable pharmaceutically acceptable carriers include, but are not limited to: (1) Dulbecco's phosphate buffered saline, pH about 7.4, with or without about 1 mg / ml to 25 mg / ml human serum albumin; (2) 0.9% saline (0.9% w / v sodium chloride), and (3) 5% (w / v) glucose; antioxidants such as tryptamine and stabilizers such as Tween 20 may also be included.

[0065] In certain embodiments, the pharmaceutically acceptable carrier of the present invention includes a pharmaceutically acceptable salt, which means that within the scope of reliable medical evaluation, the salt of the compound is suitable for contact with the tissue of humans or lower animals without undue toxicity, irritation and allergic reaction, etc., has a fairly reasonable benefit and risk ratio, is usually water or oil soluble or dispersible, and can be effectively used for its intended use. Including pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts, which can be used for the intended purpose and are compatible with the chemical properties of the compound of formula I. A list of suitable salts is, for example, see SM Birge et al., J. Pharm. Sci., 1977, 66, 1-19 pages.

[0066] The pharmaceutical composition of the present invention can be any suitable dosage form. For example, injection, suspension, emulsifier, etc. The pharmaceutical composition of the present invention can be applied to the body by known means. For example, delivered to the tissue of interest by intramuscular injection, optionally administered via intravenous, percutaneous, intranasal, oral, mucosal or other delivery methods. Such administration can be carried out via a single dose or multiple doses. It is understood by those skilled in the art that the actual dose to be administered herein can vary depending on a variety of factors to a great extent, such as target cells, biological types or their tissues, the general condition of the subject to be treated, the route of administration, the mode of administration, etc.

[0067] [In vitro methods]

[0068] In a fourth aspect of the present invention, an in vitro method for inhibiting or resisting viruses is provided, referred to herein as an "in vitro method". In addition, "in vitro" refers to an event that occurs in an artificial environment, such as in a test tube or reaction vessel, in a cell culture, in a culture dish, etc., rather than in an organism (such as an animal, plant or microorganism). Therefore, the in vitro method of the present invention is different from the method for treating a disease associated with a viral infection in a subject, which involves the action of a compound on a virus in vivo.

[0069] The in vitro method of the present invention includes the step of contacting the capsid inhibitor described in the first aspect with the virus in vitro. As long as the step of treating the virus with the inhibitor of the present invention or contacting the virus is included, the other steps are not limited and can be freely selected by those skilled in the art as needed.

[0070] The in vitro method of the present invention can be any type of in vitro test method, including but not limited to antiviral test method, compound activity test method or drug screening test method. The in vitro method of the present invention includes molecular level test method and cell level test method.

[0071] [use]

[0072] The fifth aspect of the present invention provides use of the capsid inhibitor described in the first aspect in the preparation of antiviral drugs.

[0073] The antiviral activity of the present invention includes at least one of the following activities:

[0074] The infectivity of the virus decreases or disappears;

[0075] The level of viral DNA replication decreases;

[0076] The amount of virus decreases or disappears;

[0077] The virus activity decreases or disappears;

[0078] The clinical symptoms after viral infection are alleviated or disappear.

[0079] The compounds of the present invention work by acting on the central pore of the capsid protein hexamer. Therefore, as long as the virus contains a capsid protein, especially a capsid protein hexamer, the type of virus is not limited. An exemplary virus is human immunodeficiency virus.

[0080] Example 1

[0081] This example is an example of preparing compounds ZA-25 ​​and ZA-26, which specifically includes the following steps:

[0082] Methyl 6-bromohexanoate (Z1, 3.18 g, 15.23 mmol, 1.2 eq) was added to 50 mL DMF, and then potassium carbonate (5.2 g, 38.07 mmol, 3 eq) and 2,4-thiazolidinedione (1.5 g, 12.69 mmol, 1 eq) were added, and the mixture was reacted at room temperature under magnetic stirring. After 12 h, the reaction was completed by TLC monitoring, and the solvent DMF was removed by rotary evaporation. 50 mL of ethyl acetate was added to the reaction bottle and the reaction system was transferred to a 250 mL separatory funnel, and 60 mL of water was added for extraction, and the lower organic phase was collected. The aqueous phase was washed with ethyl acetate (15 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove ethyl acetate. The crude product was mixed with 100-200 mesh silica gel and separated by column chromatography (ethyl acetate: petroleum ether = 1:8) to obtain a white oil Z2, which was directly used in the next step.

[0083] The intermediate Z2 (0.1 g, 0.4065 mmol, 1 eq) was added to 10 mL of ethanol, and then 3,4-dihydroxybenzaldehyde (56 mg, 0.4065 mmol, 1 eq) and 0.5 mL of piperidine were added, and the mixture was refluxed at 80°C under magnetic stirring. After 8 h, the reaction was completed by TLC monitoring, and 1 M hydrochloric acid was added to the reaction bottle to adjust the pH to neutral. Solid precipitated and was filtered to obtain ZA-26.

[0084] ZA-26: dark yellow solid, yield 88.7%, Mp: 128 - 130℃. 1 H NMR (400 MHz, DMSO- d 6)δ 7.74 (s, 1H, CH=), 7.06 - 6.98 (m, 2H, Ph-H), 6.89 (d, J = 8.5 Hz, 1H, Ph-H),3.63 (t, J = 7.3 Hz, 2H, CH2), 3.58 (s, 3H, OCH3), 2.30 (d, J= 16.8 Hz, 2H, CH2), 1.61 - 1.50 (m, 4H, CH2× 2), 1.30 - 1.25 (m, 2H, CH2). 13 C NMR (100 MHz, DMSO- d 6) δ 173.69 (C=O), 168.03 (C=O), 166.31 (C=O), 149.35, 146.40, 134.25,124.73, 124.63, 116.96, 116.87, 116.78, 51.65, 41.62, 33.49, 27.25, 25.81,24.41. MS: m / z 365.97 (M + H) + . C 17 H 19 NO6S [365.09].

[0085] ZA-26 (50 mg) was added to 3 mL of water and placed under magnetic stirring. Then 3 mL of acetic acid and 1 mL of water were added to the bottle and the temperature was raised to 100°C for reflux reaction. After 12 h, the reaction was completed by TLC monitoring, the solvent was removed by rotary evaporation, and then 5 mL of water was added and stirred at room temperature for 10 min, and filtered to obtain ZA-25.

[0086] ZA-25: white solid, yield 92.0 %, Mp: 174 - 175 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ7.74 (s, 1H, CH=), 7.05 - 6.97 (m, 2H, Ph-H), 6.88 (d, J = 8.1 Hz, 1H, Ph-H),3.62 (t, J = 7.2 Hz, 2H, CH2), 2.19 (t, J = 7.4 Hz, 2H, CH2), 1.62 - 1.48 (m,4H, CH2× 2), 1.26 (q, J = 6.3 Hz, 2H, CH2). 13 C NMR (100 MHz, DMSO- d6) δ 174.83(C=O), 168.02 (C=O), 166.30 (C=O), 149.32, 146.38, 134.23, 124.75, 124.63,116.97, 116.89, 116.77, 41.67, 33.87, 27.34, 26.09, 24.47. MS: m / z 351.95 (M+ H) + . C 16 H 17 NO6S [351.08].

[0087] Example 2

[0088] This example is an example for preparing compounds ZA-27 and ZA-28. The operation is the same as that of Example 1, except that m-hydroxybenzaldehyde is used.

[0089] ZA-27: white solid, yield 85.0 %, Mp: 172 - 174 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ7.82 (s, 1H, CH=), 7.15 - 7.10 (m, 2H, Ph-H), 7.02 (d, J = 11.7 Hz, 1H, Ph-H),4.34 - 4.27 (m, 4H, CH2× 2), 3.62 (t, J = 7.4 Hz, 2H, CH2), 2.29 (t, J = 7.3 Hz, 2H, CH2), 1.59 - 1.50 (m, 4H, CH2× 2), 1.31 - 1.23 (m, 2H, CH2). 13 C NMR (100MHz, DMSO- d 6) δ 174.83 (C=O), 167.86 (C=O), 166.15 (C=O), 158.38, 134.62,133.53, 130.91, 121.88, 121.58, 118.41, 116.47, 41.85, 33.87, 27.28, 26.08,24.52. MS: m / z 335.95 (M + H) + . C 16 H 17 NO5S [335.08].

[0090] ZA-28: yellow solid, yield 90.2 %, Mp: 100 - 102 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ9.88 (s, 1H, OH), 7.83 (s, 1H, CH=), 7.35 (t, J = 7.6 Hz, 1H, Ph-H), 7.05 (d, J =7.6 Hz, 1H, Ph-H), 7.00 (s, 1H, Ph-H), 6.90 (d, J = 10.8 Hz, 1H, Ph-H), 3.63(t, J = 7.2 Hz, 2H, CH2), 3.58 (s, 3H, OCH3), 2.30 (t, J = 7.4 Hz, 2H, CH2), 1.62- 1.52 (m, 4H, CH2× 2), 1.27 (dd, J = 16.5, 7.4 Hz, 2H, CH2). 13 C NMR (100 MHz, DMSO- d 6) δ 173.70 (C=O), 167.85 (C=O), 166.14 (C=O), 158.39, 134.61, 133.54,130.90, 121.86, 121.56, 118.41, 116.47, 51.65, 41.79, 33.49, 27.20, 25.98,24.40. MS: m / z 350.04 (M + H) + . C 17 H 19 NO5S [349.10].

[0091] Example 3

[0092] This example is the preparation of compounds ZA-29 and ZA-30. The operation is the same as that of Example 1, except that 1,4-benzodioxane-6-carboxaldehyde is used.

[0093] ZA-29: white solid, yield 83.9 %, Mp: 128 - 130 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ7.82 (s, 1H, CH=), 7.16 - 7.11 (m, 2H, Ph-H), 7.02 (d,J = 11.2 Hz, 1H, Ph-H),4.35 - 4.26 (m, 4H, CH2× 2), 3.62 (t, J = 7.3 Hz, 2H, CH2), 2.29 (t, J = 7.3 Hz, 2H, CH2), 1.60 - 1.52 (m, 4H, CH2× 2), 1.31 - 1.23 (m, 2H, CH2). 13 C NMR (100MHz, DMSO- d 6) δ 174.83 (C=O), 167.78 (C=O), 166.17 (C=O), 146.31, 144.22,133.26, 126.56, 124.41, 119.37, 119.22, 118.53, 64.97, 64.46, 41.80, 33.87,27.30, 26.08, 24.46. MS: m / z 377.96 (M + H) + . C 18 H 19 NO6S [377.09].

[0094] ZA-30: yellow solid, yield 91.4 %, Mp: 74 - 76 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ7.82 (s, 1H, CH=), 7.16 - 7.11 (m, 2H, Ph-H), 7.02 (d, J = 11.7 Hz, 1H, Ph-H),4.34 - 4.27 (m, 4H, CH2× 2), 3.62 (t, J = 7.2 Hz, 2H, CH2), 3.57 (s, 3H, OCH3), 2.29 (t, J = 7.4 Hz, 2H, CH2), 1.59 - 1.50 (m, 4H, CH2× 2), 1.29 - 1.23 (m, 2H, CH2). 13 C NMR (100 MHz, DMSO- d6) δ 173.69 (C=O), 167.75 (C=O), 166.16 (C=O),146.31, 144.21, 133.26, 126.73, 124.40, 119.36, 119.18, 118.51, 64.97, 64.46,51.65, 41.74, 33.73, 27.22, 25.99. MS: m / z 392.05 (M + H) + . C 19 H 21 NO6S[391.11].

[0095] Example 4

[0096] This example is an antiviral activity experiment of the target compound, which is as follows:

[0097] 1. Experimental principle:

[0098] HIV-1 pNL4-3-luc virus is generated by removing HIV Env and Vpr genes from lentiviral plasmids, into which the firefly luciferase reporter gene is inserted. This plasmid and VSVG envelope plasmid co-transfected cells can produce viruses that can infect MT4 cells. By detecting the fluorescence content in the infected cell precipitate, the degree of cell infection can be quantified. After adding the test compound to the cells, HIV-1 replication can be inhibited and the expression of the luciferase gene can be reduced. Therefore, the anti-HIV-1 activity of the compound can be reflected by quantitatively detecting the content of luciferase using a luciferase detection kit.

[0099] 2. Experimental methods:

[0100] VSVG and pNL4-3-ER-Luc plasmids were transfected into HEK293T cells. After 48 hours, the cell supernatant was collected and centrifuged at 3000 rpm for 15 min at 4 °C to remove cell debris. The supernatant was filtered through a 0.45 μM filter and stored at -80 °C for later use. 5 / well MT4 cells were added to a 96-well plate, and the specified concentration of antiviral drugs was added. At the same time, 50 μL of virus solution and 10 μg / mL polybrene were added. After 48 hours, the culture supernatant was removed by centrifugation at 1200 g, and 50 μL of cell lysis solution was added to lyse on ice for 10 min. The supernatant was centrifuged again and mixed with firefly luciferase substrate to detect the fluorescence value. The experimental results are shown in Table 1

[0101] Table 1 Anti-HIV-1 activity of compounds at the cellular level

[0102]

[0103] 3. Experimental analysis:

[0104] At the initial screening concentration of 10 μM, the lead drug ZA-15 showed a virus inhibition rate of about 70%, while the small molecules (ZA-25, ZA-26, ZA-27, ZA-28, ZA-29 and ZA-30) with structural optimization based on ZA-15 showed a virus inhibition rate of about 95%. NL4-3-luc The antiviral activity of the drugs tested by the virus showed excellent results. As shown in Table 1, ZA-29 (EC 50 =3.7±1.2 nM) and ZA-30 (EC 50 =3.8±1.3nM) was particularly effective. In addition, the R2 substituent (1, 4-benzoxazolidine>phenol>o-diphenol) had a significant effect on the antiviral activity of the compounds, with 1,4-benzoxazolidine being the dominant substituent. In addition, the antiviral activities of carboxylic acid compounds (ZA-25, ZA-27, and ZA-29) and the corresponding ester derivatives (ZA-26, ZA-28, and ZA30) were not much different, indicating that the effects of carboxyl and ester groups on the antiviral activity of the compounds were weaker than those of 1, 4-benzoxazolidine, phenol, and o-diphenol.

[0105] Although the present invention has been described with reference to exemplary embodiments, it should be understood that the present invention is not limited to the disclosed exemplary embodiments. Various adjustments or changes may be made to the exemplary embodiments of the present specification without departing from the scope or spirit of the present invention. The scope of the claims should be based on the broadest interpretation to cover all modifications and equivalent structures and functions.

Claims

1. A capsid inhibitor, characterized in that It has the structure shown in the following formula (I): Formula (I); Wherein: R1 represents a carboxyl group or a carboxylic acid methyl ester group, and R2 represents a substituted phenyl group, wherein the substituted phenyl group is selected from one of the following: 。 2. The capsid inhibitor according to claim 1, characterized in that Its structural formula is selected from one of the following: 、 、 、 、 、 。 3. The method for preparing the capsid inhibitor according to claim 1 or 2, characterized in that: The following steps are involved: (1) allowing thiazolidinedione to undergo a nucleophilic substitution reaction with a halogen-substituted alkyl ester to obtain an intermediate; (2) The intermediate is further reacted with a substituted benzaldehyde to generate an inhibitor compound.

4. The method for preparing the capsid inhibitor according to claim 3, characterized in that: The method further comprises the step of subjecting the inhibitor compound obtained in step (2) to ester hydrolysis reaction.

5. The method for preparing the capsid inhibitor according to claim 3 or 4, characterized in that: The reaction of step (2) is carried out under reflux conditions.

6. A pharmaceutical composition, characterized in that Comprising the capsid inhibitor according to claim 1 or 2 and a pharmaceutically acceptable carrier.

7. An in vitro method for inhibiting viral activity, characterized in that The method comprises the step of contacting the capsid inhibitor according to claim 1 or 2 with a virus in vitro.

8. Use of the capsid inhibitor according to claim 1 or 2 in the preparation of antiviral drugs, characterized in that: The virus comprises a capsid protein.

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