CN106928206B - Aldehyde compound and its preparation method and use - Google Patents

Abstract

The present invention relates to the field of medicinal chemistry and pharmacotherapeutics, in particular to a compound of general formula I as an enterovirus protease inhibitor, which also has significant inhibitory activity on the main protease of coronavirus (SARS), and can be used for the treatment of related diseases. The present invention also relates to methods for the preparation of such compounds, pharmaceutical compositions of these compounds, their pharmaceutically acceptable salts, enantiomeric forms, diastereoisomers and racemic compounds.

Description

Aldehyde compound and its preparation method and use

technical field

The present invention relates to the field of medicinal chemistry and pharmacotherapy, in particular to aldehyde-based compounds as inhibitors of enterovirus protease or coronavirus main protease, a preparation method thereof, pharmaceutical compositions containing such compounds and uses thereof.

Background technique

Infections caused by enteroviruses are more common in children. Its clinical characteristics are fatigue, fatigue, low-grade fever, etc. in mild patients. Severe patients can suffer from systemic infection and damage to vital organs such as the brain, heart, liver, and spinal cord. The prognosis is poor, and there are sequelae. Such diseases are distributed all over the world, with high incidence in humid and warm, poor sanitation, and densely populated areas. The infection of some viruses often occurs in epidemics, and the epidemics in different years can be caused by different types of viruses, and the epidemics of some virus infections are cyclical.

Among them, the large-scale outbreaks of hand, foot and mouth disease in the Asia-Pacific region in recent years are caused by a variety of enteroviruses, among which coxsackievirus A group 16 (CVA16) and enterovirus 71 (EV71) are the most common. At present, there is no prevention and treatment method for hand, foot and mouth disease, and only broad-spectrum antiviral drugs are used for clinical treatment. Therefore, it is a very meaningful and challenging work to develop an EV71 and CVA16 inhibitor for the treatment of hand, foot and mouth disease.

EV71 and CVA16 are highly homologous in evolution, and both viruses belong to Picronaviridae, Enterovirus, Human enterovirus A in taxonomic status. Usually the clinical symptoms caused by CVA16 infection are milder and less associated with neurological lesions; while EV71 often causes severe central nervous system diseases in addition to hand, foot and mouth disease, such as encephalitis, meningitis, aseptic meningitis, acute Flaccid paralysis, etc., can lead to pulmonary edema and heart failure in more serious cases, and the mortality rate is extremely high.

Enteroviruses are naked virions with an icosahedral structure. Because they are not coated with a lipid outer membrane, conventional disinfectants cannot effectively inactivate the virus. Although the two pathogens are sensitive to temperature and completely inactivated at 56°C in 30 minutes, the virus can survive for several weeks at 4°C, several years at -20°C, and can survive for a long time in the natural environment. Therefore, it is difficult to effectively prevent and control enterovirus infection.

Enteroviruses are single-stranded positive-sense RNAs with 5'- and 3'-untranslated regions at both ends; the viral protein coding region in the middle. The coding region contains only one open reading frame, so the original translation product of the virus is a polyprotein precursor with a molecular weight of about 243KDa. The polyprotein needs to be further cleaved by the 2A and 3C proteases encoded by the virus itself, and processed into 11 mature functional protein subunits (Vp1-Vp4, 2A-2C, 3A-3D) to complete the replication and assembly of the virus. The 2A protease is responsible for the cleavage of the Vp1/2A junction sequence; while the 3C protease is responsible for the cleavage of the other 8 sites in the polyprotein (including Vp2/Vp3, Vp3/Vp1, 2A/2B, 2B/2C, 2C/3A, 3A /3B, 3B/3C, 3C/3D junction sequence), because it can recognize multiple different sites and play a major role in the processing of viral polyprotein precursors, so the 3C protease is also called the main protease. In addition, 3C protease also has RNA binding activity. Shin-Ru13 et al. demonstrated that EV71 3C can directly participate in virus replication by binding to the 5'-UTR of the viral genome through the two functional motifs "KFRDI" and "VGK". On the other hand, 3C protease promotes virus proliferation by interacting with various host factors. For example, the host's own protein translation system is shut down by cleaving the CstF-64 host factor, thereby providing more raw materials for viral protein synthesis. Inhibits the production of the host antiviral factor interferon beta (IFN-beta) by binding to retinoic acid-inducible gene I (RIG-I). The key role of 3C in the life cycle of enteroviruses makes this protease an effective antiviral target; while humans lack the homologous protein of 3C protease, therefore, effective 3C protease inhibitors have the potential to be used in clinical, The possibility of treating related diseases caused by enterovirus infection such as hand, foot and mouth disease.

In conclusion, there is an urgent need in the art to develop 3C protease inhibitors.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a 3C protease inhibitor.

The first aspect of the present invention provides an aldehyde-based compound represented by the general formula (I), or a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof :

in,

Chiral carbon atoms C*, C* ² , C* ³ are each independently S-type, R-type, or a combination thereof;

n=0 or 1;

X is CH ₂ or NR ₅ ;

Y is a linking group selected from the group consisting of -CON(R ₄ )R ₃ -, -CH ₂ =CH ₂ -;

R ₁ is selected from the following groups unsubstituted or substituted by 1-3 substituents: C3-C7 cycloalkyl, trifluoromethyl, C2-C6 alkynyl, 4-7 membered heterocyclyl, C5-C7 Aryl, 5-7 membered heteroaryl; the heteroaryl contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; the substituents are each independently selected from the following group: halogen, C1-C4 Straight-chain or branched alkyl, C1-C4 straight-chain or branched alkenyl, C2-C4 straight-chain or branched alkynyl, C1-C4 straight-chain or branched alkoxy, C1-C4 straight-chain or branched Alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, mercapto, C1-C4 acyl, amido, sulfonyl, aminosulfonyl, C1-C4 alkyl substituted The sulfonyl group, or two adjacent substituents together with the carbon atoms attached to it form a 5-7 membered ring;

R ₂ is selected from the following groups unsubstituted or substituted by 1-3 substituents: C3-C7 cycloalkyl, 5-12-membered heterocyclic group (preferably 5-7-membered heterocyclic group or 6-membered aryl group and 5-7-membered heterocyclic group), C6-C12 aryl group, 5-12-membered heteroaryl group, or -Cbz; wherein, each of the heterocyclic group or heteroaryl group contains 1-3 selected from oxygen, sulfur and nitrogen heteroatoms; the substituents are independently selected from halogen, C1-C6 straight-chain or branched-chain alkyl, C2-C6 straight-chain or branched-chain alkenyl, C2-C6 straight-chain or branched-chain alkynyl , C1-C6 linear or branched alkoxy, C1-C6 linear or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, mercapto, C1-C4 acyl group, amide group, sulfonyl group, aminosulfonyl group, C1-C4 alkyl substituted sulfonyl group, or two adjacent substituent groups together with the carbon atoms attached to it form a 5-7 membered ring;

R ₃ is a C1-C6 alkylene group unsubstituted or substituted with 1-3 substituents; the substituents are independently selected from C1-C6 straight or branched chain alkyl, C1-C6 straight or Branched alkoxy, substituted or unsubstituted C3-C7 cycloalkyl, substituted or unsubstituted C6-C12 aryl, substituted or unsubstituted 5-12-membered heteroaryl, the heteroaryl contains 1- 3 heteroatoms selected from oxygen, sulfur and nitrogen; wherein, the cycloalkyl, aryl, and heteroaryl are replaced by one or more selected from halogen, C1-C6 straight or branched chain alkyl, cyano , nitro, amino, hydroxyl, methylol, trifluoromethyl, trifluoromethoxy, carboxyl, C1-C4 alkoxy, mercapto, C1-C4 acyl groups;

R ₄ and R ₅ are each independently selected from the group consisting of hydrogen, halogen, C1-C6 straight-chain or branched-chain alkyl, C2-C6 straight-chain or branched-chain alkenyl, C2-C6 straight-chain or branched-chain alkynyl, C3-C7 cyclic hydrocarbon group, C1-C6 acyl group, C5-C7 aryl group, benzyl group or 5-7 membered heteroaryl group; the heteroaryl group contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; wherein , the aryl group, benzyl group or 5-7 membered heteroaryl group is optionally selected from one or more halogens, C1-C6 straight or branched chain hydrocarbon groups, cyano groups, nitro groups, amino groups, hydroxyl groups, hydroxyl groups Methyl group, trifluoromethyl group, trifluoromethoxy group, carboxyl group, C1-C4 alkoxy group, mercapto group, C1-C4 acyl group are substituted.

In another preferred example, R ₁ is a substituted or unsubstituted phenyl group, preferably an unsubstituted phenyl group.

In another preferred example, R ₂ is a 5- to 6-membered heterocyclic group containing N, O or S, preferably a 5- to 6-membered heterocyclic group containing N, benzoyl.

In another preferred embodiment, in general formula (I):

Y is -CON(R ₄ )R ₃ -;

R ₁ is selected from the following groups unsubstituted or substituted with 1-3 substituents: trifluoromethyl, alkynyl, cyclopropanyl, cyclobutanyl, cyclopentyl, cyclohexyl, phenyl , thienyl, pyrazolyl, thiazolyl, pyridyl, furyl.

In another preferred embodiment, in general formula (I):

Chiral carbon atoms C*, C* ² are S type, chiral carbon atom C* ³ is S type, R type, or a combination thereof;

R ₃ is a C1-C6 alkylene group unsubstituted or substituted with 1-3 substituents; the substituents are independently selected from C1-C6 straight or branched chain alkyl, C1-C6 straight or Branched alkoxy, substituted or unsubstituted C3-C7 cycloalkyl;

R ₄ and R ₅ are each independently selected from hydrogen, C1-C4 straight-chain or branched-chain alkyl, C2-C4 straight-chain or branched alkenyl, and C2-C4 straight-chain or branched alkynyl.

In another preferred embodiment, in general formula (I):

n=1;

R ₃ is an unsubstituted or C1-C3 alkylene group substituted with 1-3 substituents; the substituents are each independently selected from the group consisting of C1-C6 straight or branched chain alkyl, substituted or unsubstituted Substituted C3-C6 cycloalkyl; said alkyl or cycloalkyl is optionally selected from one or more groups selected from halogen, C1-C4 straight or branched chain alkyl, cyano, nitro, amino, hydroxyl , hydroxymethyl, trifluoromethyl, trifluoromethoxy, carboxyl, C1-C4 alkoxy, mercapto, C1-C4 acyl groups.

In another preferred embodiment, in general formula (I):

n = 0;

X is NR ₅ ;

R ₁ is a group that is unsubstituted or substituted by 1-3 substituents and is selected from the group consisting of cyclopentyl, cyclohexyl, phenyl, thienyl;

R ₂ is a group that is unsubstituted or substituted with 1-3 substituents and is selected from the group consisting of phenyl, benzoheterocyclyl, and 5-12-membered heteroaryl; preferably, the benzoheterocycle and 5-12-membered heteroaromatic ring is selected from benzodioxole, indole, isoxazole, 2-hydropropopyran, pyridine, pyrazole, dihydroimidazopyridine, imidazopyridine, benzo Thiophene, dihydrobenzodioxane, quinoxaline, benzofuran, indazole, benzimidazole, quinoline;

R ₅ is selected from hydrogen, C1-C4 straight-chain or branched-chain alkyl, C2-C4 straight-chain or branched-chain alkenyl, C2-C4 straight-chain or branched-chain alkynyl.

In another preferred example, the aldehyde-based compound is selected from the compounds shown in Table A.

The second aspect of the present invention provides a preparation method of the compound represented by the general formula (I) as described in the first aspect of the present invention, comprising the steps:

(1) In an inert solvent, in the presence of a condensing agent, react with a compound of formula II and a compound of formula Ic to obtain a compound of formula Id; preferably, the condensing agent is EDCI (1-ethyl-(3-dimethylene) aminopropyl) carbodiimide hydrochloride);

(2) in an inert solvent, carry out a reduction reaction with a compound of formula Id to obtain a compound of formula Ie; preferably, the reduction reaction uses borohydride as a reducing agent;

(3) in an inert solvent, use the compound of formula Ie to carry out oxidation reaction to obtain the compound of formula If, namely the compound of formula (I); preferably, the oxidation reaction uses Dess-Martin oxidant or dimethyl sulfoxide and oxalyl chloride as an oxidant;

In the above formulas, the definitions of each group are as described in the first aspect of the present invention.

In another preferred example, the step (3) is carried out in the presence of a base, and the base is selected from the group consisting of sodium bicarbonate or triethylamine.

The third aspect of the present invention provides a pharmaceutical composition comprising: a therapeutically effective amount of one or more compounds represented by the general formula (I) in the first aspect of the present invention, or its Pharmaceutically acceptable salts.

The third aspect of the present invention provides a pharmaceutical composition for treating or preventing related diseases caused by enterovirus infection.

In another preferred embodiment, the pharmaceutical composition is used for inhibiting the replication of enterovirus and coronavirus; preferably, the pharmaceutical composition is used for inhibiting enterovirus 3C protease.

In another preferred embodiment, the related diseases caused by the enterovirus infection include: respiratory tract infection, herpetic angina, epidemic rash disease, hand, foot and mouth disease or meningitis.

The fourth aspect of the present invention provides the use of the compound of general formula (I) as described in the first aspect of the present invention for preparing a medicine for inhibiting the replication of enteroviruses and coronaviruses.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (eg, the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, it is not repeated here.

Detailed ways

Through long-term and in-depth research, the inventors have prepared a class of compounds of formula I that can inhibit the replication of enteroviruses and coronaviruses. And compared with the enterovirus and coronavirus replication inhibitory compounds in the prior art, the compound has higher inhibitory activity. Based on the above findings, the inventors have completed the present invention.

One object of the present invention is to provide an aldehyde-based compound represented by the general formula (I), a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof.

Another object of the present invention is to provide a preparation method of the compound represented by the above general formula (I).

Another object of the present invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of one or more compounds represented by the above general formula (I) or pharmaceutically acceptable salts thereof.

Another object of the present invention is to provide the compound represented by the above-mentioned general formula (I) in preparation for related diseases caused by enterovirus infection, such as respiratory tract infection, herpetic angina, epidemic rash, hand, foot and mouth disease, Use in medicines such as meningitis.

The compounds of the present invention can be used to inhibit the replication of enteroviruses and coronaviruses, especially to inhibit enterovirus 3C protease.

the term

As used herein, unless otherwise specified, the term "substituted" refers to the substitution of one or more hydrogen atoms on a group with a substituent selected from the group consisting of C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkane group, C ₁ -C ₁₀ alkoxy group, halogen, hydroxyl, carboxyl group (-COOH), C ₁ -C ₁₀ aldehyde group, C ₂ -C ₁₀ acyl group, C ₂ -C ₁₀ ester group, amino group, phenyl group; Said phenyl group includes unsubstituted phenyl group or substituted phenyl group with 1-3 substituents selected from: halogen, C ₁ -C ₁₀ alkyl, cyano, OH, nitro, C ₃ ~C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, amino.

Unless otherwise specified, in all compounds of the present invention, each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

The term "C1-C6 alkyl" refers to straight or branched chain alkyl groups having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or similar groups.

The term "3-8 membered heterocyclic group" refers to a group formed by the loss of a hydrogen atom from a 3- to 8-membered saturated ring having 1-3 heteroatoms selected from the group consisting of N, S, O; for example, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, or the like.

The term "6-10-membered aryl group" refers to a group formed by the loss of a hydrogen atom of a 6-10-membered aryl group; for example, a phenyl group, a naphthyl group, or the like.

The term "5-10 membered heteroaryl" refers to a group formed by the loss of one hydrogen atom of a 5-8 membered aryl group having 1-3 heteroatoms selected from the group consisting of N, S, O, wherein each heteroaryl The ring system of the radical may be monocyclic or polycyclic; for example, pyrrolyl, pyridyl, thienyl, furyl, imidazolyl, pyrimidinyl, benzothienyl, indolyl, imidazopyridyl, quinolinyl or similar groups.

The term "C1-C6 alkoxy" refers to a straight or branched chain alkoxy having 1-6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isopropoxy Butoxy, sec-butoxy, tert-butoxy, or similar groups.

The term "C2-C6 ester group" refers to a ROC(=O)- group having 2-6 carbon atoms, such as -COOCH ₃ , -COOC ₂ H ₅ , -COOC ₃ H ₇ , -COOC ₄ H ₉ , or similar groups.

The term "C2-C6 alkenyl" refers to a group formed by the loss of one or two hydrogen atoms from an olefin having 2-6 carbon atoms, which may be a mono-, di- or tri-olefin, such as -CH= _CH2 , _-C2H4 = _CH2 , _-CH = _C2H4 , or the like _.

The term "halogen" refers to F, Cl, Br and I.

Unless otherwise specified, the structural formulae described herein are intended to include all isomeric forms (such as enantiomers, diastereomers, and geometric isomers (or conformational isomers): such as those containing asymmetric centers The R, S configuration of the double bond, the (Z), (E) isomer and the (Z), (E) conformational isomer. Therefore the compound of the present invention is a single stereochemical isomer or its enantiomer Isomers, diastereomers or mixtures of geometric isomers (or conformational isomers) are within the scope of the present invention.

The term "tautomers" means that structural isomers of different energies can exceed the low energy barrier and thus interconvert. For example, proton tautomers (i.e., protonation) include interconversion by proton transfer, such as 1H-indazole with 2H-indazole, 1H-benzo[d]imidazole with 3H-benzo[d]imidazole , valence tautomers include interconversion through some bonding electron recombination.

Herein, the form "C1-C6" means that the group may have 1 to 6 carbon atoms, eg 1, 2, 3, 4 or 5 carbon atoms.

Aldehyde compounds represented by formula (I)

The present invention provides an aldehyde-based compound represented by the general formula (I), its enantiomer, diastereomer, racemate and mixture thereof or a pharmaceutically acceptable salt thereof,

Here, the definition of each group is as described above.

In another preferred embodiment, n, X, Y, R ₁ , R ₂ , R ₃ , R ₄ and R ₅ are each independently the corresponding group corresponding to each specific compound in the embodiment.

In particular, the aldehyde-based compounds of the present invention are preferably selected from the compounds shown in Table A below:

Table A

Preparation of compounds of formula (I)

The present invention also provides a method for synthesizing a compound of the general formula I, specifically, the compound of the formula I is prepared by the following scheme:

Step a: dissolving dimethyl tert-butoxycarbonyl glutamate in a solvent, adding alkali to stir at -78 ° C, then adding bromoacetonitrile, and continuing to stir to obtain compound I _a , and the solvent is tetrahydrofuran or dioxane; The alkali is lithium hexamethyldisilazide or lithium diisopropylamide;

Step b: dissolve I _a in a solvent, add catalytic amount of platinum dioxide, stir until the raw material reacts completely, filter, add alkali, reflux and stir to obtain compound I _b ; the alkali is sodium carbonate or sodium acetate; the solvent It is a mixed solvent of methanol and chloroform;

Step c: _Ib is dissolved in a solvent, stirred until the reaction is complete, and the solvent is spin-dried to obtain compound _Ic ; the solvent is a mixed solvent of dichloromethane and trifluoroacetic acid;

Step _d : the substituted carboxylic acid and _Ic are dissolved in a solvent, and a condensation reaction is carried out to obtain compound Id under the assistance of a condensing agent; the solvent is dichloromethane or DMF;

Step e: Compound I _d is dissolved in a solvent, sodium borohydride is added, stirred to obtain Compound I _e , and the solvent is methanol, tetrahydrofuran, ethanol;

Step _f : Compound _Ie is dissolved in a solvent, an oxidant is added, an alkali is added, and stirring is performed to obtain the final product If, and the solvent is dichloromethane or tetrahydrofuran; the oxidant is Dess-Martin oxidant or dimethyl sulfoxide and oxalyl chloride; the base is sodium bicarbonate or triethylamine;

X, Y, R ₁ , R ₂ are as defined in the preceding claims.

Pharmaceutical compositions containing compounds of formula (I)

The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount selected from the aldehyde compounds represented by formula (I), their pharmaceutically acceptable salts, their prodrugs and their hydrates and solvates One or more of the and optionally, pharmaceutically acceptable carriers, which can be used for the treatment of enterovirus and coronavirus replication and other related diseases. The pharmaceutical composition can be prepared in various forms according to different administration routes.

One or more of the aldehyde-based compounds represented by formula (I), their pharmaceutically acceptable salts, their prodrugs, and their hydrates and solvates according to the present invention, or the above-mentioned compounds comprising a therapeutically effective amount selected from the formula The pharmaceutical composition of one or more of the aldehyde compounds represented by (I), their pharmaceutically acceptable salts, their prodrugs and their hydrates and solvates can be used as enterovirus 3C protease inhibitors for the treatment of Enterovirus infection or illness associated with coronavirus infection.

The preparation of the pharmaceutically acceptable salts of the compounds of the present invention can be carried out by the direct salt-forming reaction of the free bases of the compounds with inorganic or organic acids. The inorganic or organic acid may be selected from hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, hydrofluoric acid, hydrobromic acid, formic acid, acetic acid, picric acid, citric acid, maleic acid, methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid acid and p-toluenesulfonic acid, etc.

Since the compound of the present invention has excellent inhibitory activity against the replication of enteroviruses and coronaviruses, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the present invention The pharmaceutical composition in which the compound of the invention is the main active ingredient can be used for the treatment, prevention and alleviation of diseases related to the replication of enteroviruses and coronaviruses, for example, the prevention and/or treatment of diseases related to abnormal replication of enteroviruses and coronaviruses. According to the prior art, the compounds of the present invention can be used to treat the following diseases: respiratory tract infection, herpetic angina, epidemic rash disease, hand, foot and mouth disease, meningitis and the like.

The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier within a safe and effective amount. The "safe and effective amount" refers to: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 5-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。"Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid filler or gelling substances which are suitable for human use and which must be of sufficient purity and sufficiently low toxicity. "Compatibility" as used herein means that the components of the composition can be admixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid) , magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween)

), wetting agents (such as sodium lauryl sulfate), colorants, flavors, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include, but are not limited to: oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and topical.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or compatibilizers, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as, for example, hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, For example, glycerol; (d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate; (e) slow solvents such as paraffin; (f) absorption Accelerators, eg, quaternary amine compounds; (g) wetting agents, eg, cetyl alcohol and glyceryl monostearate; (h) adsorbents, eg, kaolin; and (i) lubricants, eg, talc, stearin Calcium acid, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions may be in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric substances and waxes. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, and the like.

Besides these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

The compounds according to the present invention as described above can be used clinically in mammals, including humans and animals, by oral, nasal, dermal, pulmonary, or gastrointestinal routes of administration, more preferably oral administration. The daily dose is preferably 0.01-200 mg/kg body weight, taken at one time, or 0.01-100 mg/kg body weight in divided doses. Regardless of the method of administration, the optimal dose for an individual will depend on the specific treatment. It is common to start with a small dose and gradually increase the dose until the most suitable dose is found. Of course, the specific dosage should also take into account the route of administration, the patient's health and other factors, which are all within the skill of the skilled physician.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. In the following examples, the experimental methods without specific conditions are usually in accordance with conventional conditions, or in accordance with the conditions suggested by the manufacturer. Percentages and parts are by weight unless otherwise indicated.

The invention will be further illustrated in the following examples. These examples are only intended to illustrate the invention, but not to limit the invention in any way. All parameters in the examples and the rest of the description, unless otherwise stated, are based on the quality of the description.

The analytical data of the samples were determined by the following instruments: nuclear magnetic resonance was determined by GEMINI-300, Bruker AMX-400 and INVOA-600 nuclear magnetic resonance instruments, TMS (tetramethylsilane) was the internal standard, chemical shift units were ppm, coupled The unit of constant is Hz; mass spectra were determined by Finnigan type MAT-711, MAT-95 and LCQ-DECA type mass spectrometers and an IonSpec 4.7 Tesla mass spectrometer.

Silica gel 200-300 mesh for column chromatography (produced by Qingdao Ocean Chemical Plant); TLC silica gel plate is HSGF-254 thin-layer chromatography prefabricated plate produced by Yantai Chemical Plant; petroleum ether boiling range is 60-90 ℃; UV lamp is used , iodine cylinder color. Unless otherwise stated, conventional reagents and medicines used in the following examples were purchased from Sinopharm Group. The reagents and solvents used in the experiments were handled according to the specific conditions of the reaction.

Example 1: Synthesis of Compound 1

synthetic route:

Synthesis of compounds 1-3:

Under argon protection, N-tert-butoxycarbonyl-L-glutamate dimethyl ester (1-1) (6 g, 21.8 mmol) was dissolved in 60 mL of anhydrous tetrahydrofuran, and LiHMDS ( 1 M in THF) in tetrahydrofuran (47 mL, 47 mmol), and the temperature was kept stable at -78 °C during the dropwise addition for about 1 hour. After the dropping was completed, the mixture was stirred at -78°C for 1 hour. Bromoacetonitrile (2.79 g, 23.3 mmol) was dissolved in 20 ml of tetrahydrofuran, and then the solution was slowly dropped into the reaction system, and the dropwise addition process continued for 1 to 2 hours. The temperature was controlled to -78°C, and the reaction was continued for 20 hours. THL monitoring (color development of alkaline potassium permanganate) after the reaction was completed, 3 mL of methanol and 22.7 mL of a mixed solution of glacial acetic acid and tetrahydrofuran (v/v=1/7.5) were added to the reaction solution to quench the reaction, and the reaction was stirred for 10 min. to room temperature. Pour into 40 mL of saturated sodium chloride solution and stir well, and it can be seen that the reaction system is separated into layers. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (EA). The organic layers were combined, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (Flash, PE:EA=4:20) to obtain pale yellow oil 1-33.9 g, the yield is 58%.

¹ H NMR (CDCl ₃ , 400MHz) δ5.23(d, J=9.0Hz, 1H), 4.43-4.36(m, 1H), 3.77(s, 1H), 3.76(s, 1H), 2.89-2.69( m,3H),2.20-2.14(m,2H),1.45(s,9H)ppm.

ESI-MS m/z 215[M+H-boc] ⁺ .

Synthesis of compounds 1-5:

Compound 1-3 (5 g, 15.9 mmol) was dissolved in a mixed solution of 3 mL of chloroform and 50 mL of methanol, platinum dioxide (0.3 g) was added, and the mixture was stirred at room temperature under hydrogen. TLC monitoring (basic potassium permanganate), after the reaction was completed, was filtered under reduced pressure, the filtrate was evaporated to dryness, 50 mL of methanol and sodium acetate (8.46 g, 31.8 mmol) were added, and the reaction was completed in about 12 hours. The reaction solution was concentrated under reduced pressure, the residue was extracted with ethyl acetate/saturated sodium chloride, dried over anhydrous sodium sulfate, and subjected to column chromatography (Flash, PE:EA=8:20) to obtain a colorless oily substance 1-5 (2.6 g). ) with a yield of 57%.

¹ H NMR(CDCl ₃ , 400MHz)δ6.02(br,1H),5.49(d,J=7.8Hz,1H),4.33-4.27(m,1H),3.72(s,3H),3.36-3.31( m,2H),2.49-2.40(m,2H),2.16-2.06(m,1H),1.89-1.77(m,2H),1.41(s,9H)ppm.

ESI-MS m/z 309[M+Na] ⁺ .

Synthesis of compounds 1-6:

Compound 1-5 (2.6 g) was dissolved in a dichloromethane solution of trifluoroacetic acid (1/1, v/v), stirred at room temperature for 1 hour, concentrated and added with 100 ml of dichloromethane, washed with saturated sodium carbonate solution, organic The layer was dried over anhydrous sodium sulfate, and concentrated to give an oily product 1-6 (2.7 g) with a yield of 99%.

Synthesis of compounds 1-9:

Cbz-valine (1.26 g, 5 mmol), EDCI (1.36 g, 6 mmol), and HOBt (0.822 g, 6 mmol) were added to 80 ml of dichloromethane solution, and stirred at room temperature for 30 min. Subsequently, phenylalanine methyl ester (0.896 g, 5 mmol) was added, 1.2 equivalents of triethylamine were added dropwise, and the mixture was stirred at room temperature. TLC monitoring (ultraviolet) after the reaction was completed, extracted with dichloromethane, washed with dilute hydrochloric acid, saturated sodium bicarbonate solution and saturated sodium chloride, combined the organic layers, dried over anhydrous sodium sulfate, and concentrated to obtain 1.8 g of a white viscous solid , the yield is 90%.

Synthesis of compounds 1-10:

1-9 (1.8 g, 4.36 mmol) was dissolved in 80 ml of methanol, 0.2 g of Pd/C was added, and the mixture was stirred at room temperature under a hydrogen atmosphere. The reaction was complete after about 24 hours. The reaction solution was suction filtered through diatomaceous earth, and concentrated to obtain compound 1-101.15 g with a yield of 94.7%.

Synthesis of compounds 1-13:

Acid 1-11 (5 mmol), EDCI (1.36 g, 6 mmol), HOBt (0.822 g, 6 mmol) were added to 30 ml of dichloromethane solution, and stirred at room temperature for 30 min. Then 1-10 (5 mmol) was added, 1.2 equivalents of triethylamine were added dropwise, and the mixture was stirred at room temperature. TLC monitoring (ultraviolet) after the reaction is complete, extract with dichloromethane, wash with dilute hydrochloric acid, saturated sodium bicarbonate solution and saturated sodium chloride, combine the organic layers, dry with anhydrous sodium sulfate, and concentrate to obtain a white oily substance, which can be directly used for next reaction.

The oily intermediate obtained in the previous step was dissolved in a mixed solution of methanol/water (1:2, v/v), 3 equivalents of 1M aqueous sodium hydroxide solution were added, and the mixture was stirred at room temperature. After TLC monitoring (ultraviolet) the reaction was completed, the pH was adjusted to -2, extracted with ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried with anhydrous sodium sulfate, and concentrated to obtain 1.4 g of white solid with a yield of 61.5%.

Synthesis of compounds 1-14:

Dissolve 1-13 (592 mg, 1.3 mmol) and intermediate 1-6 (242 mg, 1.3 mmol) in 20 ml of DMF solution, then add condensing agent HATU (592 mg, 1.6 mmol) and 1.2 equivalents of triethylamine, and stir at room temperature . After TLC monitoring (ultraviolet) the reaction was completed, the mixture was extracted with ethyl acetate/saturated ammonium chloride solution, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic layer was concentrated. After separation by flash column chromatography (CH ₂ Cl ₂ :MeOH=30:1), 1-14320 mg of light yellow oily product was obtained, with a yield of 39.5%.

Synthesis of compounds 1-15:

1-14 (320 mg, 0.51 mmol) was dissolved in 20 ml of methanol, sodium borohydride (107 mg, 2.9 mmol) was slowly added in batches, and the reaction was completed by stirring at room temperature for about 2 hours. After the reaction was completed, about 20 ml of saturated brine was added to quench the reaction, and after the methanol was concentrated in the reaction system, dichloromethane was added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain light yellow oily substance 1-15, which could be directly used in the next reaction.

Synthesis of compound 1:

Intermediate 1-15 (173 mg, 0.29 mmol) was dissolved in 20 ml of dichloromethane, and Dess-Martin oxidant (147 mg, 0.35 mmol) and solid sodium bicarbonate (29 mg, 0.35 mmol) were added at room temperature. Stir. After TLC monitoring (ultraviolet) the reaction was completed, the reaction system was suction filtered, the obtained filtrate was dissolved in saturated sodium bicarbonate for extraction, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated. After separation and purification by flash column chromatography (CH ₂ Cl ₂ :MeOH=20:1), a total of 62 mg of compound 1 was obtained as a white solid powder with a yield of 35.9%.

¹ H NMR (400MHz, CDCl3): δ9.45-9.36 (m, 1H), 7.76 (d, J=7.6Hz, 1H), 7.54-7.18 (m, 8H), 7.03-6.98 (m, 1H), 6.54-6.18(m, 1H), 5.18-4.90(m, 1H), 4.61(m, 2H), 4.49-4.14(m, 2H), 3.48-3.13(m, 4H), 2.65(m, 1H), 2.44-2.28(m,1H),2.26-2.17(m,1H),2.02-1.64(m,3H),0.92(m,6H).ESI-MS m/z 595.22[M+H] ⁺ .

Example 2: Synthesis of Compound 2

The acid 1-13 in Example 1 is replaced by compound 1-11, and the synthesis method refers to the synthesis of compound 1 to obtain compound 2. ¹ H NMR (400MHz, CDCl3): δ9.32-9.26 (m, 1H), 7.81 (m, 1H), 7.54-7.18 (m, 8H), 7.13-6.91 (m, 1H), 6.44-6.08 (m ,1H),5.15-4.90(m,1H),4.65(m,2H),4.47-4.04(m,2H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16 (m,1H),2.12-1.54(m,3H).ESI-MSm/z 496.15[M+H] ⁺ .

Example 3: Synthesis of Compound 3

Substitute compound 3-1 for 1-11 in Example 1, and the synthesis method refers to the synthesis of compound 1 to obtain compound 3. ¹ H NMR (400MHz, CDCl3): δ9.52-9.43 (m, 1H), 7.54-7.25 (m, 5H), 6.47-6.18 (m, 2H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,2H),3.45-3.03(m,4H),2.65(m,1H),2.54-2.30(m,4H),2.24-2.16(m,1H),2.12-1.54(m,3H),0.92 (m,6H).ESI-MS m/z 512.24[M+H] ⁺ .

Example 4: Synthesis of Compound 4

Substitute compound 4-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 4. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 9H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H) .ESI-MS m/z 447.20[M+H] ⁺ .

Example 5: Synthesis of Compound 5

Substitute compound 5-1 for 1-11 in Example 1, and the synthesis method refers to the synthesis of compound 1 to obtain compound 5. ¹ H NMR (400MHz, CDCl3): δ9.52-9.39(m,1H), 7.64-7.17(m,8H), 6.47-6.18(m,1H), 6.070(s,2H), 5.14-4.96(m) ,1H),4.45-4.04(m,2H),3.45-3.03(m,4H),2.65(m,1H),2.54-2.30(m,1H),2.25-2.08(m,1H),2.12-1.44 (m,3H),0.96(m,6H).ESI-MS m/z551.24[M+H] ⁺ .

Example 6: Synthesis of Compound 6

Substitute compound 4-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 6. ¹ H NMR (400MHz, CDCl3): δ9.46-9.34 (m, 1H), 7.54-7.25 (m, 9H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,2H),3.45-3.03(m,4H),2.65(m,1H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12 -1.54(m,3H),0.92(m,6H).ESI-MS m/z546.26[M+H] ⁺ .

Example 7: Synthesis of Compound 7

Substitute compound 5-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 7. ¹ H NMR (400MHz, CDCl3): δ9.52-9.41 (m, 1H), 7.64-7.17 (m, 8H), 6.47-6.18 (m, 1H), 6.07 (s, 2H), 5.14-4.96 (m ,1H),4.45-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.25-2.08(m,1H),2.12-1.44(m,3H).ESI -MS m/z 452.17[M+H] ⁺ .

Example 8: Synthesis of Compound 8

Substitute compound 8-1 for 1-11 in Example 1, and the synthesis method refers to the synthesis of compound 1 to obtain compound 8. ¹ H NMR(400MHz, CDCl3): δ9.58-9.42(m,1H), 8.78(s,1H), 7.92(s,1H), 7.54-7.25(m,5H), 6.47-6.18(m,1H) ), 5.15-4.96(m, 1H), 4.47-4.04(m, 2H), 3.91(s, 3H), 3.45-3.03(m, 4H), 2.65(m, 1H), 2.54-2.30(m, 1H ),2.24-2.16(m,1H),2.12-1.54(m,3H),0.94(m,6H).ESI-MSm/z 511.26[M+H] ⁺ .

Example 9: Synthesis of Compound 9

Substitute compound 9-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 9. ¹ H NMR (400MHz, CDCl3): δ9.48-9.36(m,1H), 8.78(s,1H), 8.68(s,1H), 7.92(s,1H), 7.54-7.15(m,5H), 6.47-6.18(m, 1H), 5.15-4.96(m, 1H), 4.47-4.04(m, 2H), 3.45-3.03(m, 4H), 2.65(m, 1H), 2.54-2.30(m, 1H) ),2.24-2.16(m,1H),2.12-1.54(m,3H),0.94(m,6H).ESI-MSm/z 526.24[M+H] ⁺ .

Example 10: Synthesis of Compound 10

Substitute compound 9-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 10. ¹ H NMR (400MHz, CDCl3): δ9.50-9.45(m, 1H), 8.80(s, 1H), 8.65(s, 1H), 7.89(s, 1H), 7.54-7.15(m, 5H), 6.47-6.18(m, 1H), 5.15-4.96(m, 1H), 4.47-4.04(m, 1H), 3.45-3.03(m, 4H), 2.54-2.30(m, 1H), 2.24-2.16(m ,1H),2.12-1.54(m,3H).ESI-MS m/z427.17[M+H] ⁺ .

Example 11: Synthesis of Compound 11

Substitute compound 8-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 11. ¹ H NMR(400MHz, CDCl3): δ9.58-9.42(m,1H), 8.78(s,1H), 7.92(s,1H), 7.54-7.25(m,5H), 6.47-6.18(m,1H) ), 5.15-4.96(m, 1H), 4.47-4.04(m, 1H), 3.87(s, 3H), 3.45-3.03(m, 4H), 2.54-2.30(m, 1H), 2.24-2.16(m ,1H),2.12-1.54(m,3H).ESI-MS m/z427.17[M+H] ⁺ .

Example 12: Synthesis of Compound 12

Synthesis of compound 12-3:

D-phenylalanine (5 g, 30.3 mmol) was dissolved in 60 mL of 48% hydrobromic acid, cooled to 0 °C, and sodium nitrite (2.6 g, 37.5 mmol) was added in batches, and the temperature was stirred for 1 hour. At room temperature, the reaction was continued for 10 hours. The reaction was completed, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain 5.75 g of colorless oily compound 12-3 with a yield of 83%.

Synthesis of compound 12-4:

Compound 12-3 (4 g) was dissolved in 30 mL of methanol, 1 mL of concentrated sulfuric acid was added, heated, and the temperature was raised to 70° C. to react for 2 hours. Concentrate and evaporate methanol, add 10 mL of water, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and column chromatography to obtain colorless oil 12-4, 3.95 g, yield 93%.

Synthesis of compound 12-5:

Dissolve L-benzyloxycarbonylvaline (3 g, 11.9 mmol) in 15 mL of anhydrous tetrahydrofuran, under argon protection, add 1,1'-dicarbonyl imidazole (CDI, 2.13 g, 13.1 mmol), and stir at room temperature for 1 hour , cooled to -78 ℃, set aside. The lithium diisopropylamide solution (2 equiv. LDA solution in tetrahydrofuran, 19 mL) was diluted with 10 mL of tetrahydrofuran, protected with argon, cooled to -78°C, added with tert-butyl acetate (4.87 g, 42 mmol), and stirred for 1 hour. The reaction solution of L-benzyloxycarbonylvaline and CDI was added, the temperature was controlled to -78°C, and the reaction was continued for 2 hours. 2 mL of water was added to quench, warmed to room temperature, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain 2.1 g of yellow oil 12-5 with a yield of 60%.

Synthesis of compound 12-6:

Compound 12-4 (2 g, 8.23 mmol) was dissolved in 30 mL of THF, compound 12-5 (5.75 g, 16.5 mmol), potassium carbonate (2.56 g, 18.54 mmol), tetrabutylammonium iodide (200 mg) were added, The reaction was carried out at room temperature for 36 hours. The reaction solution was poured into 60 mL of water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain compound 12-61.26 g with a yield of 30%.

Synthesis of compound 12-7:

Compound 12-6 (1.26 g, 2.47 mmol) was dissolved in 15 mL of a mixed solution of trifluoroacetic acid and dichloromethane (v/v=3/1), and the temperature was controlled at 30° C. to react for 4 hours. Concentrated, added 10 mL of water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain 760 mg of compound 12-7 with a yield of 75%.

Synthesis of compounds 12-8:

Compound 12-7 (0.76 g, 1.85 mmol) was dissolved in 20 mL of methanol, 10% Pd-C (80 mg) was added,

Air was expelled, hydrogen gas was introduced, and the mixture was stirred at room temperature for 10 hours. Suction filtration and concentration to obtain 0.44 g of compound 12-8 with a yield of 85%.

Synthesis of compound 12:

The acid 1-11 in Example 1 was replaced with compound 8-1, and the compound 12-8 was replaced with 1-10 in Example 1. The synthesis method was referred to the synthesis of compound 1, and compound 12 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.56-9.41(m,1H), 8.79(s,1H), 7.90(s,1H), 7.54-7.18(m,5H), 6.47-6.18(m,1H) ), 5.15-4.96(m, 1H), 4.47-4.04(m, 2H), 3.91(s, 3H), 3.45-3.03(m, 5H), 2.65-2.57(m, 2H), 2.54-2.30(m ,1H),2.24-2.16(m,1H),2.12-1.54(m,3H),0.94(m,6H).ESI-MS m/z 510.26[M+H] ⁺ .

Example 13: Synthesis of Compound 13

Substitute compound 5-1 for 8-1 in Example 12, and the synthesis method refers to the synthesis of compound 12 to obtain compound 13. ¹ H NMR (400MHz, CDCl3): δ9.52-9.41 (m, 1H), 7.64-7.17 (m, 8H), 6.47-6.18 (m, 1H), 6.08 (s, 2H), 5.15-4.96 (m ,1H),4.47-4.04(m,2H),3.45-3.03(m,5H),2.65-2.57(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12 -1.54(m,3H),0.94(m,6H).ESI-MS m/z 550.25[M+H] ⁺ .

Example 14: Synthesis of Compound 14

Substitute compound 14-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 14. ¹ H NMR (400MHz, CDCl ₃ )δ9.72-9.08(m,1H),8.34-8.21(m,2H),8.16-8.10(m,1H),8.06-7.94(m,2H),7.42(ddd , J=15.1, 6.6, 4.7Hz, 2H), 7.29 (dd, J=13.9, 2.0Hz, 2H), 7.18–7.12 (m, 2H), 6.51 (dd, J=36.8, 13.4Hz, 1H), 5.04 (tt, J=15.2, 7.6Hz, 1H), 4.47–4.18 (m, 1H), 3.24 (dd, J=14.3, 7.5Hz, 4H), 2.24–2.16 (m, 1H), 2.01–1.83 ( m,3H),1.76–1.60(m,2H).ESI-MS m/z 526.10[M+H] ⁺ .

Example 15: Synthesis of Compound 15

Substitute compound 15-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 15.

¹ H NMR (400MHz, CDCl ₃ ) δ 9.40 (dd, J=53.9, 44.1 Hz, 1H), 8.34-8.18 (m, 1H), 7.40-7.28 (m, 3H), 7.26-7.19 (m, 4H) ), 6.87 (dd, J=8.4, 2.5Hz, 1H), 6.80–6.61 (m, 1H), 5.69 (dd, J=9.7, 6.1Hz, 1H), 5.05–4.92 (m, 1H), 4.36 ( dt,J=8.5,5.1Hz,5H),3.49(s,1H),3.34-3.24(m,3H),3.20-3.09(m,1H),2.35-2.25(m,1H),1.86(dddd, J=20.6,17.0,13.2,4.2Hz,3H).ESI-MS m/z 466.19[M+H] ⁺ .

Example 16: Synthesis of Compound 16

Substitute compound 16-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 16.

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.68-9.22 (m, 1H), 8.61-8.18 (m, 1H), 7.92-7.67 (m, 4H), 7.44-7.35 (m, 2H), 7.31-7.27 (m, 2H), 7.25–7.15 (m, 2H), 7.13–6.99 (m, 1H), 6.08–5.81 (m, 1H), 5.12–4.97 (m, 1H), 4.48–4.21 (m, 1H) ,3.50–3.13(m,5H),2.38–2.16(m,2H),2.02–1.84(m,2H).ESI-MS m/z 464.16[M+H] ⁺ .

Example 17: Synthesis of Compound 17

Substitute compound 17-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 17.

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.56-9.25 (m, 1H), 8.23-7.99 (m, 2H), 7.68-7.61 (m, 1H), 7.28 (dd, J=7.4, 6.1 Hz, 3H ), 7.26–7.20 (m, 2H), 7.01–6.94 (m, 1H), 6.89 (dd, J=15.2, 7.3Hz, 1H), 6.04 (dd, J=78.3, 34.4Hz, 1H), 5.03 ( dq, J=10.5, 7.3Hz, 1H), 4.34–4.23 (m, 5H), 3.50–3.34 (m, 1H), 3.32–3.23 (m, 3H), 3.18–3.08 (m, 1H), 2.37– 2.21(m,1H),2.00–1.85(m,2H),1.77–1.69(m,1H).ESI-MS m/z 466.19[M+H] ⁺ .

Example 18: Synthesis of Compound 18

Substitute compound 18-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 18. ¹ H NMR (400MHz, CDCl ₃ ) δ 9.61-9.48(m, 1H), 9.46-9.20(m, 1H), 8.63-8.42(m, 2H), 8.24-7.93(m, 2H), 7.89-7.78 (m,2H),7.34-7.27(m,3H),7.24-7.17(m,1H),6.54-6.18(m,1H),5.18-4.90(m,1H),4.49-4.14(m,1H) ,3.48-3.13(m,4H),2.44-2.28(m,1H),2.26-2.17(m,1H),2.02-1.64(m,3H).ESI-MS m/z 460.19[M+H] ⁺ .

Example 19: Synthesis of Compound 19

Substitute compound 19-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 19.

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.53-9.19 (m, 1H), 8.45 (ddd, J=17.0, 10.0, 5.9 Hz, 1H), 7.94-7.68 (m, 1H), 7.65 (dd, J =7.8,0.8Hz,1H),7.51–7.37(m,4H),7.31–7.27(m,4H),7.22(dd,J=9.3,4.8Hz,1H),6.32–5.97(m,1H), 5.10–5.01 (m, 1H), 4.47–4.07 (m, 2H), 3.51–3.34 (m, 1H), 3.32–3.18 (m, 4H), 2.37–2.21 (m, 1H), 1.92–1.83 (m ,2H).ESI-MS m/z 448.18[M+H] ⁺ .

Example 20: Synthesis of Compound 20

Substitute compound 20-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 20. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 8.32-7.52 (m, 4H), 7.48-7.25 (m, 5H), 6.35-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H) .ESI-MS m/z 448.19[M+H] ⁺ .

Example 21: Synthesis of Compound 21

Substitute compound 21-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 21. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 8.72-7.82 (m, 5H), 7.42-7.25 (m, 5H), 6.35-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H) .ESI-MS m/z 460.19[M+H] ⁺ .

Example 22: Synthesis of Compound 22

Substitute compound 22-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 22.

¹ H NMR (400MHz, CDCl ₃ )δ12.79-12.45(m,1H), 9.53-9.29(m,1H), 9.11-8.80(m,1H), 8.54-8.17(m,2H), 8.01-7.68 (m, 2H), 7.55–7.42 (m, 1H), 7.35–7.27 (m, 3H), 7.22–7.15 (m, 2H), 6.57 (dd, J=27.5, 23.5Hz, 1H), 5.24–4.99 (m, 1H), 4.35–4.22 (m, 1H), 3.48–3.23 (m, 3H), 2.98–2.60 (m, 2H), 2.25 (ddd, J=13.0, 12.4, 5.9Hz, 1H), 2.14 –2.06(m,1H),1.94–1.81(m,2H).ESI-MS m/z 448.19[M+H] ⁺ .

Example 23: Synthesis of Compound 23

Substitute compound 23-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 23.

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.54-9.26 (m, 1H), 8.43 (ddd, J=11.5, 10.8, 4.1 Hz, 1H), 8.04 (dddd, J=36.5, 9.5, 5.8, 1.9 Hz ,2H),7.74–7.60(m,1H),7.45–7.27(m,5H),7.25–7.14(m,2H),7.03–6.82(m,1H),6.61(s,1H),6.00(d , J=53.6Hz, 1H), 5.15-5.00(m, 1H), 4.49-4.22(m, 1H), 3.36-3.19(m, 3H), 2.35-2.20(m, 2H), 2.05-1.95(m ,2H),1.84–1.72(m,2H).ESI-MS m/z 447.19[M+H] ⁺ .

Example 24: Synthesis of Compound 24

Substitute compound 24-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 24. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 10H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.99(s,3H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H).ESI -MS m/z 461.21[M+H] ⁺ .

Example 25: Synthesis of Compound 25

Substitute compound 25-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 25. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 9H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H).ESI-MS m/z 465.19[ M+H] ⁺ .

Example 26: Synthesis of Compound 26

Substitute compound 26-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 26. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 9H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.28(s,3H),2.24-2.16(m,1H),2.12-1.54(m,3H).ESI -MS m/z 461.21[M+H] ⁺ .

Example 27: Synthesis of Compound 27

Substitute compound 27-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 27. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 9H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.83(s,3H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H).ESI -MS m/z 477.21[M+H] ⁺ .

Example 28: Synthesis of Compound 28

Substitute compound 28-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 28. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 8H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,6H).ESI-MS m/z 462.20[ M+H] ⁺ .

Example 29: Synthesis of Compound 29

Substitute compound 29-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 29. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.72 (s, 1H), 7.52-7.18 (m, 8H), 6.35-6.18 (m, 1H), 5.15-4.96 (m ,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H).ESI -MS m/z 481.16[M+H] ⁺ .

Example 30: Synthesis of Compound 30

Substitute compound 30-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 30. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 8.85-7.87 (m, 6H), 7.52-7.18 (m, 5H), 6.35-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H) .ESI-MS m/z 459.20[M+H] ⁺ .

Example 31: Synthesis of Compound 31

Substitute compound 31-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 31. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 8.98-7.96 (m, 5H), 7.52-7.18 (m, 5H), 6.35-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H) .ESI-MS m/z 527.18[M+H] ⁺ .

Example 32: Synthesis of Compound 32

Substitute compound 32-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 32. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 8.31-7.86 (m, 5H), 7.52-7.18 (m, 5H), 6.35-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H) .ESI-MS m/z 537.11[M+H] ⁺ .

Example 33: Synthesis of Compound 33

Substitute compound 33-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 33. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.82-7.64 (m, 4H), 7.52-7.18 (m, 5H), 6.35-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.63(s,3H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12 -1.54(m,3H).ESI-MS m/z474.21[M+H] ⁺ .

Example 34: Synthesis of Compound 34

Substitute compound 18-1 for 1-11 in Example 2, and substitute compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 34. ¹ H NMR (400MHz, CDCl ₃ ) δ 9.61-9.48 (m, 1H), 9.46-9.20 (m, 1H), 8.63-8.42 (m, 2H), 8.24-7.93 (m, 2H), 7.24-7.17 (m,1H),6.35-6.18(m,1H),5.15-4.96(m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H) ,2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H).ESI-MS m/z466.24[M+H] ⁺ .

Example 35: Synthesis of Compound 35

Substitute compound 18-1 for 1-11 in Example 2, and substitute compound 35-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 35. ¹ H NMR (400MHz, CDCl ₃ ) δ 9.61-9.48 (m, 1H), 9.46-9.20 (m, 1H), 8.63-8.42 (m, 2H), 8.24-7.93 (m, 2H), 7.24-7.17 (m,1H),6.35-6.18(m,1H),5.15-4.96(m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H) ,2.24-2.16(m,1H),2.12-1.57(m,5H),1.55-1.35(m,9H).ESI-MS m/z452.22[M+H] ⁺ .

Example 36: Synthesis of Compound 36

Substitute compound 5-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 36. ¹ H NMR (400MHz, CDCl3): δ9.54-9.47(m,1H), 7.64-7.17(m,3H), 6.37-6.18(m,1H), 6.07(s,2H), 5.15-4.96(m) ,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62 -1.41(m,11H).ESI-MS m/z 458.22[M+H] ⁺ .

Example 37: Synthesis of Compound 37

Compound 15-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 37 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.51-9.43 (m, 1H), 7.68-7.13 (m, 3H), 6.37-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,5H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H) .ESI-MS m/z471.24[M+H] ⁺ .

Example 38: Synthesis of Compound 38

Compound 17-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 38 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.55-9.47 (m, 1H), 7.68-7.13 (m, 3H), 6.32-6.11 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,5H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H) .ESI-MS m/z471.24[M+H] ⁺ .

Example 39: Synthesis of Compound 39

Substitute compound 39-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 39. δ9.51-9.46(m,1H),7.56-8.12(m,4H),7.25-7.43(m,5H),6.35-6.13(m,1H),5.14-4.95(m,1H),4.46-4.04 (m,1H), 3.44-3.02(m,4H), 2.51-2.31(m,1H), 2.27-2.18(m,1H), 2.12-1.89(m,3H), 1.79(s,3H).ESI -MS m/z 462.21[M+H] ⁺ .

Example 40: Synthesis of Compound 40

Substitute compound 40-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 40. ¹ H NMR (400MHz, CDCl3): δ9.51-9.46 (m, 1H), 7.59-8.17 (m, 4H), 7.25-7.51 (m, 5H), 6.37-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.75-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.27-1.52(m,3H) .ESI-MS m/z 466.18[M+H] ⁺ .

Example 41: Synthesis of Compound 41

Substitute compound 41-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 41. ¹ H NMR (400MHz, CDCl3): δ9.49-9.43 (m, 1H), 7.59-8.18 (m, 4H), 7.21-7.49 (m, 5H), 6.37-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.75-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.27-1.52(m,3H) .ESI-MS m/z 482.15[M+H] ⁺ .

Example 42: Synthesis of Compound 42

Substitute compound 42-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 42. δ9.50-9.44(m,1H),7.52-8.13(m,4H),7.18-7.47(m,5H),6.37-6.13(m,1H),5.15-4.95(m,1H),4.48-4.04 (m,1H),3.51(s,3H),3.44-3.02(m,4H),2.51-2.31(m,1H),2.27-2.18(m,1H),2.12-1.89(m,3H).ESI -MS m/z 478.20[M+H] ⁺ .

Example 43: Synthesis of Compound 43

Compound 39-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 43 was obtained. δ9.51-9.46(m,1H),7.45-8.07(m,4H),6.35-6.13(m,1H),5.14-4.95(m,1H),4.47-4.04(m,1H),3.45-3.03 (m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.79(s,3H),1.62-1.41(m,11H).ESI -MS m/z468.25[M+H] ⁺ .

Example 44: Synthesis of Compound 44

Substitute compound 42-1 for 1-11 in Example 2, and substitute compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 44. δ9.50-9.44(m,1H),7.45-8.07(m,4H),6.37-6.13(m,1H),5.15-4.95(m,1H),4.47-4.04(m,1H),3.51(s ,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H).ESI -MSm/z 484.25[M+H] ⁺ .

Example 45: Synthesis of Compound 45

Substitute compound 14-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 45. ¹ H NMR (400MHz, CDCl3): δ9.56-9.42 (m, 1H), 7.42-8.13 (m, 4H), 6.37-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H) .ESI-MS m/z532.15[M+H] ⁺ .

Example 46: Synthesis of Compound 46

Substitute compound 46-1 for 1-11 in Example 2, and the synthesis method refers to the synthesis of compound 2 to obtain compound 46. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.55-7.13 (m, 8H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,3H).ESI-MS m/z 515.12[ M+H] ⁺ .

Example 47: Synthesis of Compound 47

Compound 24-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 47 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 5H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.99(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62 -1.41(m,11H).ESI-MS m/z 467.26[M+H] ⁺ .

Example 48: Synthesis of Compound 48

Substitute compound 48-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 48. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 4H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.99(s,3H),3.81(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54 (m,5H),1.62-1.41(m,11H).ESI-MS m/z 497.26[M+H] ⁺ .

Example 49: Synthesis of Compound 49

Substitute compound 24-1 for 1-11 in Example 2, and compound 49-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 49. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.55-7.13 (m, 9H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.99(s,3H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,4H),2.12-1.54(m,3H).ESI -MSm/z 475.23[M+H] ⁺ .

Example 50: Synthesis of Compound 50

Compound 26-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 50 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.52-7.18 (m, 4H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.28(s,3H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62 -1.41(m,11H).ESI-MS m/z 467.26[M+H] ⁺ .

Example 51: Synthesis of Compound 51

Compound 46-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 51. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 7.55-7.13 (m, 3H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H) .ESI-MS m/z521.44[M+H] ⁺ .

Example 52: Synthesis of Compound 52

Compound 4-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 52. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H) ,1.62-1.41(m,11H).ESI-MS m/z 453.24[M+H] ⁺ .

Example 53: Synthesis of Compound 53

Compound 3-1 was used to replace 1-11 in Example 2, and compound 35-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 53 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.58-9.42(m,1H), 6.23(m,1H), 6.47-6.18(m,1H), 5.15-4.96(m,1H), 4.47-4.04(m ,1H),3.87(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.57(m,5H),1.55-1.35 (m,9H).ESI-MS m/z 405.21[M+H] ⁺ .

Example 54: Synthesis of Compound 54

Substitute compound 3-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 54. ¹ H NMR (400MHz, CDCl3): δ9.58-9.42(m,1H), 6.23(m,1H), 6.47-6.18(m,1H), 5.15-4.96(m,1H), 4.47-4.04(m ,1H),3.87(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41 (m,11H).ESI-MSm/z 419.23[M+H] ⁺ .

Example 55: Synthesis of Compound 55

Compound 1-1 was used to replace 1-11 in Example 2, and compound 35-1 was used to replace 1-8 of Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 55 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.32-9.26 (m, 1H), 7.81 (m, 1H), 7.54-7.18 (m, 2H), 7.13-6.91 (m, 1H), 6.44-6.08 (m ,1H),5.15-4.90(m,1H),4.65(m,2H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16 (m,1H),2.12-1.57(m,5H),1.55-1.35(m,9H).ESI-MS m/z 488.19[M+H] ⁺ .

Example 56: Synthesis of Compound 56

Compound 30-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 56 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.50-9.42 (m, 1H), 8.85-7.87 (m, 6H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H) .ESI-MS m/z465.24[M+H] ⁺ .

Example 57: Synthesis of Compound 57

Compound 57-1 is used to replace 1-11 in Example 2, and compound 34-1 is used to replace 1-8 in Example 2. The synthesis method is referred to the synthesis of compound 2 to obtain compound 57. ¹ H NMR (400MHz, CDCl3): δ9.52-9.45 (m, 1H), 8.80-7.65 (m, 6H), 6.35-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.62-1.41(m,11H) .ESI-MS m/z465.24[M+H] ⁺ .

Example 58: Synthesis of Compound 58

Substitute compound 18-1 for 1-11 in Example 2, and compound 58-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 58. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.61-9.48 (m, 1H), 9.46-9.20 (m, 1H), 8.63-8.42 (m, 2H), 8.24-7.93 (m, 3H), 7.89-7.78 (m,2H),7.34-7.27(m,4H),7.24-7.17(m,2H),6.54-6.18(m,1H),5.18-4.90(m,1H),4.49-4.14(m,1H) ,3.48-3.13(m,4H),2.44-2.28(m,1H),2.26-2.17(m,1H),2.02-1.64(m,3H).ESI-MS m/z 466.15[M+H] ⁺ .

Example 59: Synthesis of Compound 59

Compound 4-1 was used to replace 1-11 in Example 2, and compound 59-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 59. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,12H) .ESI-MS m/z425.21[M+H] ⁺ .

Example 60: Synthesis of Compound 60

Compound 4-1 was used to replace 1-11 in Example 2, and compound 60-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 60 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H) ,0.5-0.7(m,5H).ESI-MS m/z 411.20[M+H] ⁺ .

Example 61: Synthesis of Compound 61

Compound 4-1 was used to replace 1-11 in Example 2, and compound 61-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2, and compound 61 was obtained. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H) ,1.62-1.41(m,10H).ESI-MS m/z 471.23[M+H] ⁺ .

Example 62: Synthesis of Compound 62

Compound 4-1 was used to replace 1-11 in Example 2, and compound 62-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 62. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H) ,1.32-1.49(m,9H).ESI-MS m/z 489.22[M+H] ⁺ .

Example 63: Synthesis of Compound 63

Compound 4-1 was used to replace 1-11 in Example 2, and compound 63-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 63. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H) ,1.34-1.53(m,9H).ESI-MS m/z 489.22[M+H] ⁺ .

Example 64: Synthesis of Compound 64

Substitute compound 4-1 for 1-11 in Example 2, and compound 64-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 64. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 7.03-7.18 (m, 3H), 6.99-6.85 (m, 2H), 6.47-6.18 (m,1H),5.15-4.96(m,1H),4.47-4.04(m,1H),3.45-3.03(m,4H),2.54-2.30(m,1H),2.24-2.16(m,1H) ,2.12-1.54(m,3H).ESI-MS m/z 483.18[M+H] ⁺ .

Example 65: Synthesis of Compound 65

Compound 65-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 65. ¹ H NMR (400MHz, CDCl3): δ9.31-9.46 (m, 1H), 7.36-7.89 (m, 3H), 6.47-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.93(s,3H),3.87(s,3H),3.83(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m ,1H),2.12-1.54(m,5H),1.34-1.53(m,11H).ESI-MS m/z 543.27[M+H] ⁺ .

Example 66: Synthesis of Compound 66

Substitute compound 4-1 for 1-11 in Example 2, and compound 66-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 66. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.54-7.25 (m, 4H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,3H),2.24-2.16(m,1H),2.12-1.54(m,3H) .ESI-MS m/z438.15[M+H] ⁺ .

Example 67: Synthesis of Compound 67

Substitute compound 67-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 67. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.56-7.25 (m, 3H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.45-3.03(m,2H),2.54-2.30(m,4H),2.24-2.16(m,1H),2.12-1.54(m,5H) ,1.34-1.53(m,11H).ESI-MS m/z 467.26[M+H] ⁺ .

Example 68: Synthesis of Compound 68

Substitute compound 68-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 68. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.56-7.25 (m, 3H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.83(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12 -1.54(m,5H),1.34-1.53(m,11H).ESI-MS m/z 483.25[M+H] ⁺ .

Example 69: Synthesis of Compound 69

Substitute compound 69-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 69. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.46-7.25 (m, 2H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.93(s,3H),3.83(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16 (m,1H),2.12-1.54(m,5H),1.34-1.53(m,11H).ESI-MS m/z 513.26[M+H] ⁺ .

Example 70: Synthesis of Compound 70

Compound 70-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 70. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.46-7.25 (m, 2H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.96(s,3H),3.81(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16 (m,1H),2.12-1.54(m,5H),1.34-1.53(m,11H).ESI-MS m/z 513.26[M+H] ⁺ .

Example 71: Synthesis of Compound 71

Compound 71-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 71. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.46-7.25 (m, 2H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.92(s,3H),3.85(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16 (m,1H),2.12-1.54(m,5H),1.34-1.53(m,11H).ESI-MS m/z 513.26[M+H] ⁺ .

Example 72: Synthesis of Compound 72

Substitute compound 72-1 for 1-11 in Example 2, and compound 34-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 72. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.46-7.25 (m, 2H), 6.99-6.85 (m, 2H), 6.47-6.18 (m, 1H), 5.15-4.96 (m,1H),4.47-4.04(m,1H),3.94(s,3H),3.81(s,3H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16 (m,1H),2.12-1.54(m,5H),1.34-1.53(m,11H).ESI-MS m/z 513.26[M+H] ⁺ .

Example 73: Synthesis of Compound 73

Compound 73-1 was used to replace 1-11 in Example 2, and compound 34-1 was used to replace 1-8 in Example 2. The synthesis method was referred to the synthesis of compound 2 to obtain compound 73. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.46-7.25 (m, 3H), 6.47-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,2H),2.54-2.30(m,1H),2.24-2.16(m,1H),2.12-1.54(m,5H),1.34-1.53(m,11H) .ESI-MS m/z494.20[M+H] ⁺ .

Example 74: Synthesis of Compound 74

Substitute compound 18-1 for 1-11 in Example 2, and substitute compound 74-1 for 1-8 in Example 2. The synthesis method refers to the synthesis of compound 2 to obtain compound 74. ¹ H NMR (400MHz, CDCl3): δ9.46-9.31 (m, 1H), 7.46-7.25 (m, 4H), 6.47-6.18 (m, 1H), 5.15-4.96 (m, 1H), 4.47-4.04 (m,1H),3.45-3.03(m,3H),2.54-2.30(m,3H),2.24-2.16(m,1H),2.12-1.54(m,3H).ESI-MSm/z 408.16[M +H] ⁺ .

Activity Test Experiment

Example 1: EV71-3C protease inhibition experiment

Determination of compound inhibitory activity against EV71-3C protease: Purified 3C protease and chemically synthesized fluorescent substrate polypeptides were used. The coding sequence of EV71 virus 3C protease was cloned into pET-21a vector, and a hexahistidine tag was introduced at the C-terminus of the protein through the vector and expressed by E.coli; the protease was purified by Ni-NTA affinity chromatography and Superdex200 Size exclusion chromatography was completed. The substrate polypeptide will use Dabcyl-KIGNTIEALFQGPPKFRE-Edans fluorescent polypeptide, the polypeptide sequence corresponds to the junction sequence between EV71 2C/3A, the detection of polypeptide cleavage is excited by 340nm excitation light and monitored by 490nm emission light. After each synthetic compound was mixed with 3C protease at a gradient concentration, the inhibition of polypeptide cleavage efficiency was determined; through three independent experiments, the half-inhibitory concentration IC ₅₀ of each compound on 3C protease was calculated. The experimental results are shown in Table 1.

Table 1: EV71 3C protease inhibitory activity

Experimental conclusion: It can be seen from the above experimental results that the compound has a good inhibitory effect on EV71 3C protease, especially the inhibitory activity of compounds 3, 4, 7, 9, and 13 is less than 1 μM

Example 2: Evaluation of SARS coronavirus main protease inhibitory activity

Determination of the inhibitory activity of compounds against SARS (Severe Acute Respiratory Syndrom) coronavirus main protease (SARS-CoV M ^pro ): The enzyme-level inhibitory activity of the inhibitors against 3C protease was determined by fluorescence resonance energy transfer (FRET) technology. In a 96-well plate, 27.5 μL of buffer (20 mM Tris, 100 mM NaCl, 1 mM EDTA, pH 7.4) was added to each well, along with 2.5 μL of compound (final concentrations of 2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 12 μM, 14 μM, 16 μM, 18 μM, 20 μM) and 5 μL EV713Cpro (final concentration 3 μM). Incubate for 15 min at 37°C. Then 15 μL of buffer diluted fluorescent substrate (final concentration 20 μM) was added. Fluorescence parameters were measured with a Gen5 fluorescence spectrometer, the excitation wavelength and emission wavelength were 340 nm and 490 nm, respectively, kept at 37 °C, and the data was read after 10 min. A negative control was used, in which no compound was added to the control, and the rest were the same. The obtained data were processed using the software GraphPad Prism 5, and the experimental results are shown in Table 2.

Table 2: SARS coronavirus main protease inhibitory activity

Numbering IC₅₀(μM) compound IC₅₀(μM) 1 5.15 16 0.32 2 7.4 17 0.78 3 0.75 18 0.56 4 0.03 19 1.69 5 0.7 20 4.64 6 0.05 twenty one 1.13 7 0.1 twenty two 5.4 8 0.6 twenty three 2.62 9 0.035 twenty four 4.21 10 0.2 25 0.111 11 0.45 26 0.79 12 1.05 AG7088 6.7 13 1.10

Experimental conclusion: It can be seen from the above experimental results that the compounds have a good inhibitory effect on the main protease of SARS coronavirus, among which, the inhibitory activity of most compounds is far better than that of AG7088, indicating that the compounds of the present invention may be used for SARS coronavirus. Treatment of diseases related to viral main protease activity or expression.

Example 3: Evaluation of CVB3 3C protease inhibitory activity

Determination of the inhibitory activity of compounds against CVB3 (Coxsachievirus B3) 3C protease: 20.0 μL of buffer (20 mM Tris, 100 mM NaCl, 1 mM EDTA, 10 mM DTT, pH 7.4) was added to each well in a 96-well plate, and 2.5 μL of compound was added at the same time (final concentrations of 100 μM, 33 μM, 11 μM, 3.7 μM, 1.2 μM, 0.4 μM, 0.13 μM, 0.004 μM, respectively) and 2.5 μL of CVB3 3Cpro (3 μM final concentration). Incubate for 10 min at 37°C. Then add 25 μL of buffer diluted fluorogenic substrate. The fluorescence parameters were measured with a Gen5 fluorescence spectrometer. The excitation wavelength and emission wavelength were 340 nm and 490 nm, respectively, and the data was read after 15 min at 37 °C. A negative control was used, in which no compound was added to the control, and the rest were the same. The obtained data was processed using the software GraphPad Prism 6.0, and the experimental results are shown in Table 3.

Table 3: CVB3 3C protease protease inhibitory activity

Numbering IC₅₀(μM) compound IC₅₀(μM) 3 4.11 13 6.39 4 5.94 17 1.13 5 4.76 18 1.36 6 2.32 19 1.28 7 2.61 20 0.95 8 2.64 twenty one 2.67 9 2.50 twenty two 1.12 10 3.66 twenty three 1.87 11 6.10 25 1.69 12 4.43 26 0.92

Experimental conclusion: It can be seen from the above experimental results that the compound has a good inhibitory effect on CVB3 3C protease, and the IC ₅₀ can reach about 1 μM.

Example 4: Evaluation of Compound Inhibitory Activity for Each Virus Replication

Determination of the compound's inhibitory activity on the replication of each virus: 100 μl/well of gradient concentration of compound was added to 96 wells, followed by 50 μl/well of virus buffer, followed by 50 μl/well of cultured RD cells (rhabdomyosarcoma cells) immediately, at 37°C Incubate for 3-4 days until maximum cytopathic effect is observed. Aspirate the medium, add 75 μl of 5% MTS phenol red medium, incubate at 37°C, 5% CO ₂ for 1.5 hours, measure the fluorescence value of each well at the wavelength of 498 nM, and draw a graph of compound concentration and cell response, which is used in Accelrys' custom software calculates the _EC50 values of the compounds.

Table 4: Evaluation of compound inhibitory activity against EV71-BRCR virus replication

Experimental conclusion: It can be seen from the above experimental results that the compounds have a good inhibitory effect on EV71-BRCR virus replication, and the activity EC ₅₀ of some compounds is less than 1μmol.

Table 5: Evaluation of compound inhibitory activity against EV68-WT virus replication

Some compounds were selected to test their inhibitory effect on EV68-WT virus replication. From the above experimental results, it can be seen that the compounds have a good inhibitory effect on EV68-WT virus replication, and the activity reaches nanomolar level.

Table 6: Evaluation of Compounds for Inhibitory Activity of MNV (Murine norovirus)-CW1 Virus Replication

Experimental conclusion: It can be seen from the above experimental results that the compounds of the present invention also have a certain inhibitory effect on MNV-CW1 virus replication, and the activity of some compounds is better than that of the positive compound AG7088, the EC ₅₀ of compounds 56 and 48 is less than 1 μmol, and the therapeutic index TI>20.

Example 5: In vivo pharmacokinetic study of compound 18 in rats

1. Experimental steps:

6 healthy rats, weighing 150-200 g, were randomly divided into 2 groups with 3 rats in each group. The compound 18 of the present invention was administered by gavage and intravenous injection respectively, and the dosage was 10 mg/kg, and the administration volume was 10 mL/kg. ) preparation. The rats were fasted for 12 h before the test and had free access to water. 2h after the administration of unified food.

2. Blood collection time point and sample processing:

Subcutaneous administration: 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 hours after administration;

Intravenous administration: 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 hours after administration;

According to the above set time points, 0.3 mL of blood was collected from the retroocular venous plexus of the rat, placed in a heparinized test tube, centrifuged at 11,000 rpm for 5 min, and the plasma was separated and frozen in a -20°C refrigerator.

3. Sample testing and data analysis

The concentration of compound 18 in rat plasma was determined by LC/MS/MS method.

The non-compartmental model of WinNonlin 5.3 software (Pharsight, USA) was used to calculate the pharmacokinetic parameters after administration.

4. Experimental results

Table 7: Results of pharmacokinetic experiments of compound 18 in rats

After intravenous injection of 10 mg/kg of the compound 18 of the present invention in rats, the time to peak plasma concentration Tmax was 1 h, and the peak concentration Cmax was 1919.4 ng/ml; the area under the drug-time curve AUC0-t was 6876.0 ng·h/ml; terminal elimination Half-life t1/2 is 1.35h. After intravenous injection of 10 mg/kg of compound 18 of the present invention, AUC0-t was 23361.2 ng·h/ml; after dose normalization, the absolute bioavailability of compound 18 after subcutaneous injection of 10 mg/kg in rats was 29.4%.

Experimental conclusion: It can be seen from the above experimental results that in the pharmacokinetic experiments in rats, compound 18 showed good pharmacokinetic properties.

All documents mentioned herein are incorporated by reference in this application as if each document were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (18)

1. an aldehyde-based compound represented by general formula (I) or a pharmaceutically acceptable salt thereof:

in, Chiral carbon atoms C*, C* ² , C* ³ are each independently S-type, R-type, or a combination thereof; n = 0; X is NR ₅ ; Y is: -CON(R ₄ )R ₃ -; R ₁ is selected from the following groups unsubstituted or substituted by 1-3 substituents: C5-C7 aryl, 5-7 membered heteroaryl; the heteroaryl contains 1-3 selected from oxygen, sulfur and nitrogen heteroatoms; the substituents are each independently selected from the group consisting of halogen, C1-C4 linear or branched alkyl, C1-C4 linear or branched alkoxy, C1-C4 linear or Branched-chain alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, methylol, trifluoromethyl, carboxyl; R ₂ is selected from the following groups unsubstituted or substituted by 1-3 substituents: 5-12-membered heteroaryl, and the 5-12-membered heteroaryl is selected from the following group: indole, isooxane azole, pyridine, pyrazole, imidazopyridine, benzothiophene, quinoxaline, benzofuran, indazole, benzimidazole, quinoline; the substituents are independently selected from halogen, C1-C6 straight chain Or branched alkyl, C1-C6 linear or branched alkoxy, C1-C6 linear or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl , carboxyl, mercapto, C1-C4 acyl, aminosulfonyl, C1-C4 alkyl substituted sulfonyl; R ₃ is CH ₂ unsubstituted or substituted with 1-3 substituents; the substituents are independently selected from C1-C6 straight-chain or branched-chain alkyl groups; R ₄ , R ₅ are each independently selected from the group consisting of hydrogen. 2. aldehyde compound as claimed in claim 1 or its pharmaceutically acceptable salt, it is characterized in that, in general formula (I): R ₁ is selected from the following groups unsubstituted or substituted with 1-3 substituents: phenyl, thienyl, pyrazolyl, thiazolyl, pyridyl, furyl. 3. aldehyde-based compound according to claim 2, its pharmaceutically acceptable salt, is characterized in that, in general formula (I), R ₁ is a group selected from the group consisting of phenyl, thienyl, unsubstituted or substituted with 1-3 substituents. 4. The aldehyde-based compound according to claim 1, or a pharmaceutically acceptable salt thereof:

5. An aldehyde-based compound represented by general formula (I), or a pharmaceutically acceptable salt thereof:

in, Chiral carbon atoms C*, C* ² , C* ³ are each independently S-type, R-type, or a combination thereof; n=1; X is CH ₂ or NR ₅ ; Y is -CON(R ₄ )R ₃ -; R ₁ is selected from the following groups unsubstituted or substituted by 1-3 substituents: C3-C7 cycloalkyl, trifluoromethyl, C2-C6 alkynyl, 4-7 membered heterocyclyl, C5-C7 Aryl, 5-7 membered heteroaryl; each of the heterocyclic and heteroaryl contains 1 to 3 heteroatoms selected from oxygen, sulfur and nitrogen; the substituents are each independently selected from the following group: Halogen, C1-C4 linear or branched alkyl, C1-C4 linear or branched alkoxy, C1-C4 linear or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxyl Methyl, trifluoromethyl, carboxyl; R ₂ is selected from the following groups unsubstituted or substituted by 1-3 substituents: C6-C12 aryl, 5-12-membered heteroaryl; wherein, each of the heteroaryls contains 1-3 selected from heteroatoms of oxygen, sulfur and nitrogen; the substituents are each independently selected from halogen, C1-C6 straight or branched alkyl, C1-C6 straight or branched alkoxy, C1-C6 straight or Branched-chain alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, mercapto, C1-C4 acyl, aminosulfonyl, C1-C4 alkyl substituted sulfonyl; R ₃ is CH ₂ unsubstituted or substituted with 1-3 substituents; the substituents are independently selected from C1-C6 straight-chain or branched-chain alkyl groups; R ₄ and R ₅ are each independently selected from the group consisting of hydrogen, C1-C6 straight or branched chain alkyl. 6. aldehyde compound as claimed in claim 5 or its pharmaceutically acceptable salt is characterized in that, in general formula (I): R ₁ is selected from the following groups unsubstituted or substituted with 1-3 substituents: trifluoromethyl, alkynyl, cyclopropanyl, cyclobutanyl, cyclopentyl, cyclohexyl, phenyl , thienyl, pyrazolyl, thiazolyl, pyridyl, furyl. 7. aldehyde-based compound as claimed in claim 5, its pharmaceutically acceptable salt, is characterized in that, in general formula (I): Chiral carbon atoms C*, C* ² are S type, chiral carbon atom C* ³ is S type, R type, or a combination thereof; R ₃ is an unsubstituted or C1-C6 alkylene group substituted with 1-3 substituents; the substituents are independently selected from C1-C6 straight or branched chain alkyl groups; R ₄ and R ₅ are each independently selected from hydrogen, C1-C4 straight-chain or branched-chain alkyl. 8. The aldehyde-based compound and pharmaceutically acceptable salt thereof according to claim 5, wherein the 5-12-membered heteroaryl is selected from the group consisting of indole, isoxazole, pyridine, pyridine azole, imidazopyridine, benzothiophene, quinoxaline, benzofuran, indazole, benzimidazole, quinoline. 9. The aldehyde compound according to claim 5, or a pharmaceutically acceptable salt thereof:

10. An aldehyde compound represented by general formula (I), or a pharmaceutically acceptable salt thereof:

in, Chiral carbon atoms C*, C* ² , C* ³ are each independently S-type, R-type, or a combination thereof; n = 0; X is NR ₅ ; Y is -CON(R ₄ )R ₃ -; R ₁ is selected from the following groups unsubstituted or substituted with 1-3 substituents: C3-C7 cycloalkyl, trifluoromethyl, C2-C6 alkynyl, 4-7 membered heterocyclic group; the heterocyclic group The cyclic group contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; the substituents are each independently selected from the following group: halogen, C1-C4 straight or branched chain alkyl, C1-C4 straight chain or Branched alkoxy, C1-C4 linear or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, methylol, trifluoromethyl, carboxyl; R ₂ is selected from the following groups unsubstituted or substituted by 1-3 substituents: 5-12-membered heteroaryl, and the 5-12-membered heteroaryl is selected from the following group: indole, isooxane azole, pyridine, pyrazole, imidazopyridine, benzothiophene, quinoxaline, benzofuran, indazole, benzimidazole, quinoline; the substituents are independently selected from halogen, C1-C6 straight chain Or branched alkyl, C1-C6 linear or branched alkoxy, C1-C6 linear or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl , carboxyl, mercapto, C1-C4 acyl, aminosulfonyl, C1-C4 alkyl substituted sulfonyl; R ₃ is CH ₂ unsubstituted or substituted with 1-3 substituents; the substituents are independently selected from C1-C6 straight-chain or branched-chain alkyl groups; R ₄ , R ₅ are each independently selected from the group consisting of hydrogen. 11. the aldehyde compound as claimed in claim 10, or its pharmaceutically acceptable salt, is characterized in that, in general formula (I): R ₁ is selected from the following groups unsubstituted or substituted with 1-3 substituents: trifluoromethyl, cyclopropanyl, cyclobutanyl, cyclopentyl, cyclohexyl. 12. The aldehyde-based compound according to claim 10, and its pharmaceutically acceptable salt, characterized in that, in the general formula (I), R ₁ is an unsubstituted or 1-3 substituent substituted option. Groups from the group: cyclopentyl, cyclohexane. 13. The aldehyde-based compound according to claim 10, or a pharmaceutically acceptable salt thereof:

14. a preparation method of the compound shown in general formula (I) as claimed in claim 1,5 or 10, comprising the steps:

(1) in an inert solvent, in the presence of a condensing agent, react with a compound of formula II and a compound of formula Ic to obtain a compound of formula Id;

(2) in an inert solvent, carry out a reduction reaction with a compound of formula Id to obtain a compound of formula Ie;

(3) in inert solvent, carry out oxidation reaction with formula Ie compound, obtain formula If compound, namely formula (I) compound; In the above formulas, the definition of each group is as described in claim 1, 5 or 10. 15. The method of claim 14, wherein in step (1), the condensing agent is 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride; In step (2), described reduction reaction uses borohydride as reducing agent; In step (3), the oxidation reaction uses Dess-Martin oxidant or dimethyl sulfoxide and oxalyl chloride as the oxidant. 16. A pharmaceutical composition, characterized in that, the pharmaceutical composition comprises: a therapeutically effective dose of one or more compounds shown in the general formula (I) of claim 1, 5 or 10, or its pharmacy acceptable salt. 17. The use of the general formula (I) according to claim 1, 5 or 10, characterized in that it is used to prepare a pharmaceutical composition for treating or preventing related diseases caused by enterovirus infection. 18. the purposes of general formula (I) as claimed in claim 1,5 or 10, is characterized in that, is used for preparing the medicine that suppresses enterovirus and/or coronavirus replication.