WO2006042478A1 - Small-molecule inhibitors of coronaviral main protease, their preparation and use - Google Patents

Abstract

The present invention is directed to a series of small-molecule inhibitors of general formula (I), which are designed on the crystal structure of SARS coronaviral main protease. Preparation of the said small-molecule inhibitors, pharmaceutical composition containing the same as well as their use in the manufacture of a medicament for treating or preventing a variety of coronaviral infections are also provided.

Description

 Small molecule inhibitor of coronavirus main protease, preparation method thereof and application thereof

Technical field

 The present invention provides a series of small molecule inhibitors designed based on the crystal structure of SARS coronavirus main protease, a preparation method thereof, a pharmaceutical composition comprising the same, and use thereof for preparing a medicament for treating or preventing various coronavirus infections . Background technique

 A new type of coronavirus has been identified as the culprit of the Severe Acute Respiratory Sydrome (SARS) and is named SARS Coronavirus (abbreviated as SARS-CoV). The SARS coronavirus belongs to the genus "Nidovirales", "Coronaviridae", and "Coronavirus" in the species classification. It is a new variant of the coronavirus family, with a total length of 29,736 bp (Urbani Strain) o SARS is a potent infectious disease that poses a great threat to humans. There are no specific drugs and vaccines available.

 According to serological classification, the coronavirus family (Coronavirus) includes four types (Group). Type I includes porcine transmissible gastroenteritis virus (TGEV), human coronavirus (HCoV) strain 229E, and feline infectious peritonitis virus (FIPV). Type II includes Bovine coronavirus (BCoV), Murine hepatitis virus (MHV), etc. Type III currently includes only three viruses, one of which is called avian infectious. Avian Infectious Bronchitis Virus (abbreviated as AIBV); SARS-CoV belongs to type IV. Various coronaviruses pose a huge threat to the health of people and animals.

The genome of the SARS coronavirus encodes two large replicase polyproteins (replicase polyproteins) ppla (486 kDa) and pplab (790 kDa), these two proteins are encoded by the genome of 2/3 to 3/4 of the coronavirus. These two proteins are hydrolyzed to produce many functional subunits of the viral replication complex. In this hydrolysis process, the main proteolytic enzyme of SARS coronavirus (main protease, abbreviated as M ^pro , molecular weight 33.8 kDa, sometimes called 3C-Like Protein) plays a very important role. The main protease is also a segment of ppla and pplab, its release is achieved by autocatalytic hydrolysis, and autocatalytic hydrolysis occurs at the Gln S _er , Al _a ) site beside the protease, by trans-splicing (bi-molecular reaction) carry out. Under the action of the main protease, the replicase polyproteins ppla and pplab are hydrolyzed into more than a dozen functional peptides, which further play a role. If it can inhibit the hydrolysis of SARS coronavirus main protease, it will effectively resist the infection of SARS coronavirus. Therefore, SARS coronavirus primary protease is an ideal target for anti-SARS drug design. Summary of invention

 It is an object of the present invention to provide a small molecule inhibitor capable of effectively inhibiting the activity of a coronavirus main protease.

 Another object of the present invention is to provide a process for the preparation of the small molecule inhibitor.

 It is still another object of the present invention to provide a pharmaceutical composition comprising the small molecule inhibitor. It is still another object of the present invention to provide the use of the small molecule inhibitor for the preparation of a medicament for the treatment or prevention of a coronavirus infection.

 Thus, in one aspect, the invention provides a compound of formula (I): or a pharmaceutically acceptable salt or solvate thereof:

其中，

among them,

， 其中 X为 NH或 CH₂; R,选自 由如下基团构成的组： 任选被卤素取代的（^〜0₆ (优选 C₃〜C₆) 垸 基羰基、 C₃〜C₆环垸基羰基、 任选被 C₆〜C₁₍₎芳基取代的（^〜05垸 氧羰基、 C₆〜 。芳基羰基和任选被 (^〜 ₄垸基取代的 C₅〜C_W杂芳 基羰基； R₂选自由如下基团构成的组：其中芳基任选被卤素取代的任 选被 C₆〜C_1Q芳基取代的 〜C₆ (优选 〜C₄)烷基和 C₆〜C_1Q芳基； R₃选自由如下基团构成的组： 其中芳基任选被卤素取代的任选被 C₆〜C_{1 ()}芳基取代的 〜C₆ (优选 C,〜C₄) 垸基、 任选被（^〜( ₄烷 基取代的 C₆〜C_1Q芳基； 选自由如下基团构成的组： 其中芳基任选 被卤素取代的任选被 C₆〜C_{1 Q}芳基取代的 〜C₆(优选 (^〜( 垸基、 任选被 〜C₄垸基取代的 C,。芳基； R₅选自由如下基团构成的 组： 其中芳基任选被卤素取代的任选被 C₆〜C_{1 ()}芳基取代的 （^〜 ₆ (优选 (^〜( ₄) 垸基、 任选被 〜C₄垸基和 /或卤素取代的 C₆〜C₁₀ 芳基。 U is

Wherein X is NH or CH _2; R, groups selected from the group consisting of: optionally substituted by halogen ₍₆ ~ 0 ^ (preferably C ₃ ~C ₆₎ alkyl with a carbonyl group, C ₃ ~C ₆ cycloalkyl embankment a carbonyl group, optionally substituted by a C ₆ ～C ₁₍₎ aryl group (^~05垸 oxycarbonyl, C ₆ 〜. arylcarbonyl, and optionally C ₅ ～C _W substituted by (^~ ₄ fluorenyl) Arylcarbonyl; R _{2 is} selected from the group consisting of: -C ₆ (preferably -C ₄ )alkyl and C ₆ wherein the aryl group is optionally substituted by halogen, optionally substituted by C ₆ -C _1Q aryl ~C _1Q aryl; R _{3 is} selected from the group consisting of: -C ₆ (preferably C, ~C _{4 )} wherein the aryl group is optionally substituted by halogen, optionally substituted by C ₆ -C _{1 ()} aryl a fluorenyl group, optionally (^~( ₄ alkyl-substituted C ₆ -C _1Q aryl; group selected from the group consisting of: wherein the aryl group is optionally substituted by halogen, optionally C ₆ -C _{1 Q} aryl substituted ~C ₆ (preferably (^~( fluorenyl, C, optionally substituted by ~C ₄ fluorenyl); R _{5 is} selected from the group consisting of: wherein aryl is optionally Halogen substituted optionally substituted by C ₆ ～C _{1 ()} aryl (^~ ₆ (preferably (^~( ₄ ) Embankment, optionally substituted alkyl with ~C ₄ and / or halogen-substituted C ₆ ~C ₁₀ aryl group.

 In another aspect, the invention also provides a process for the preparation of a compound of formula (I), which comprises the steps of:

(a) removing a protecting group R ₆ of an amino group in the compound of the formula (II), which is selected from the group consisting of a group of ( ₆ to ₆ alkoxycarbonyl groups optionally substituted by a C ₆ -C _1Q aryl group; Optionally substituted by halogen (^~( ₆ alkyl acyl, especially tert-butoxycarbonyl, trifluoroacetyl or benzyloxycarbonyl;

(b) condensing the product of step (a) with a compound of formula (III) in the presence of a condensing reagent to provide a compound of formula (I),

 (I)

Wherein, the definitions of the formulas, R ₂ , R 4 and U are the same as defined in the above formula (I). In another aspect, the invention provides a pharmaceutical composition comprising a compound of the above formula (I), or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carriers.

 In another aspect, the invention also provides the use of a compound of formula (I) for the manufacture of a medicament for the treatment or prevention of a coronavirus infection.

 The test proves that the compound of the present invention can significantly inhibit the activity of the main protease of coronavirus such as TGEV, HCoV, FIPV, AIBV, SARS-CoV, and has a good application prospect in the preparation of a medicament for treating or preventing coronavirus infection. BRIEF DESCRIPTION OF THE DRAWINGS

 The drawings are not necessarily to scale unless the The invention may be better understood by considering the drawings in conjunction with the specific embodiments.

Figure 1 is a surface view of the small molecule inhibitor N1 of the present invention combined with SARS-CoV M ^pro monomer A;

Figure 2 is a graph showing the activity of the small molecule inhibitor N1 of the present invention and the SARS-CoV M ^pro monomer A Electronic density map of sexual pockets;

Figure 3 shows the small molecule inhibitors Nl, N2, N3 and N4 of the present invention versus SARS-CoV M ^p r . Inhibitory activity curve, wherein SARS-3CL represents SARS coronavirus main protease, ie SARS-CoV M ^p r° ;

 Figure 4 is a graph showing the inhibitory activity curve of the small molecule inhibitor N1 of the present invention against the main protease of Transmissible Gastroenteritis Virus (TGEV) (abbreviated as TGEV-3CL);

 Figure 5 is a graph showing the inhibitory activity curve of the small molecule inhibitor N1 of the present invention against the human coronavirus (HCoV) 229 main protease (abbreviated as HCoV-3CL in the figure);

 Figure 6 is a graph showing the inhibitory activity curve of the small molecule inhibitor N1 of the present invention against Feline Infectious Peritonitis Virus (FIPV) main protease (abbreviated as FIPV_3CL in the figure);

 Fig. 7 is a graph showing the inhibitory activity curve of the small molecule inhibitor N1 of the present invention against the Avian Infectious Bronchitis Virus (AIBV) main protease (abbreviated as AIBV-3CL). Detailed description of the invention

Definition of Terms

 For the convenience of the description, some specific terms are used herein, which are explained one by one below.

"Nl", "N2", "N3" and "N4" are particularly preferred small molecule inhibitors of the present invention, the structural formulas of which are:

 Structural formula of N4

The terms "main proteolytic enzymes of SARS coronavirus", "SARS-CoV M ^pro ", "SARS-CoV 3CL ^pro ", "SARS coronavirus main protease", etc., are used herein to refer to the main SARS coronavirus. Proteolytic enzymes.

"C,~C ₆ fluorenyl" means a straight or branched alkyl group having from 1 to 6 carbon atoms, which includes but is not limited to: methyl, ethyl, n-propyl, isopropyl , n-butyl, isobutyl Base, tert-butyl, n-pentyl, n-hexyl, and the like.

 "~0^alkyl" means a straight or branched fluorenyl group having from 1 to 4 carbon atoms, which includes: methyl, ethyl, n-propyl, isopropyl, n-butyl, Isobutyl, tert-butyl and the like.

The "C ₃ -C ₆ cycloalkyl group" means a cyclic alkyl group having 3 to 6 carbon atoms, and includes: a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group and the like.

"C ₆ -C _1Q aryl" means a hetero atom-free (monocyclic and bicyclic) aromatic ring having from 6 to 10 carbon atoms, which includes: phenyl, naphthyl such as 1-naphthyl or 2-naphthyl.

The "C ₅ -C _1Q heteroaryl group" means an aromatic 5- to 10-membered (monocyclic or bicyclic) heterocyclic ring containing 1 to 3 hetero atoms each selected from an oxygen atom, a sulfur atom and a nitrogen atom, and includes: Oxazolyl, isoxazolyl, furyl, imidazolyl, pyridyl, quinolyl, isoquinolyl and benzimidazolyl, and the like.

 "Halogen" means fluorine, chlorine, bromine and iodine.

 "Fluorobenzyl" includes p-fluorobenzyl, m-fluorobenzyl, o-fluorobenzyl and the like.

 "Fluorophenyl" includes p-fluorophenyl, m-fluorophenyl, o-fluorophenyl and the like.

 "Methylphenyl" includes p-methylphenyl, m-methylphenyl, o-methylphenyl and the like. Abbreviation

 In the partial structural formula, Pr" represents an isopropyl group, "Et" represents an ethyl group, "Βι represents a phenyl group, and "Boc" represents a tert-butoxycarbonyl group.

"TFA" stands for trifluoroacetic acid, "THF" stands for tetrahydrofuran; "DMF" stands for Ν, Ν-dimethylformamide; "DMSO" stands for dimethyl sulfoxide; "PhH" stands for benzene; "ipr ₂ NEt" stands for two Isopropylethylamine; "NEt ₃ " stands for triethylamine; "DCC" stands for dicyclohexylcarbodiimide; "DIEA" stands for diisopropylethylamine; "DMAP" stands for 4-Ν,Ν-dimethyl Aminopyridine; "EDCI" stands for 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride; "HATU" stands for N-[(dimethylamino)(3//-1 , 2,3-triazolo(4,5-b)pyridin-3-oxy)methylene]methylammonium hexafluorophosphate ( N- [(dimethylamino)(3 - 1 ,2,3-triazolo(4,5-b)pyridine-3-yloxy)methyle ne]-iV-methylmethanaminium hexafluorophosphate ); " HBTU " stands for ( benzotriazole - 1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (0-(benzotriazol- 1 -yl)-1,1,3,3-tetramethyluronium hexafluorophosphate); "HOBt" stands for 1 - Hydroxybenzotriazole. Structure of the compound of the present invention

 As described above, the structure of the compound of the present invention is as shown in the formula (I). , and X is NH; or U

， 且 X为 NH; 或者 U为 ， 且 X为 CH: 也就是说， 本发明的一些优选化合物符合下述通式： for

And X is NH; or U is, and X is CH: that is, some preferred compounds of the invention conform to the following formula:

 (la)

 Further preferred compounds of the invention conform to the general formula:

 ( lb )

Further preferred compounds of the invention are in accordance with the following formula:

更优选地， 在通式 （I) 中， 选自由如下基团构成的组： 三氟 甲基羰 、 叔丁氧羰基、 异噁唑基羰基、 呋喃基

More preferably, in the formula (I), a group consisting of the following groups is selected: trifluoromethylcarbonyl, tert-butoxycarbonyl, isoxazolylcarbonyl, furyl

R₂为甲基、 异丙基、 苯基、 苄基、 氟 代苄基； R₃选自由如下基团构成的组： 甲基、 异丙基、 苄基、 氟代苄 基 （特别是对氟苄基）、 苯基和甲基苯基 （特别是对甲基苯基）； R4 选自由如下基团构成的组： 乙基、 苯基、 苄基、 甲基苯基（特别是对 甲基苯基）和氟代苄基； R₅为异丁基、 苯基、 苄基、 甲基苯基（特别 是对甲基苯基）、 氟代苄基 （特别是对氟苄基） 和氟代苯基。 Carbonyl,

R ₂ is methyl, isopropyl, phenyl, benzyl, fluorobenzyl; R _{3 is} selected from the group consisting of methyl, isopropyl, benzyl, fluorobenzyl (especially Fluorobenzyl), phenyl and methylphenyl (especially p-methylphenyl); R4 is selected from the group consisting of ethyl, phenyl, benzyl, methylphenyl (especially Rhenyl) and fluorobenzyl; R ₅ is isobutyl, phenyl, benzyl, methylphenyl (especially p-methylphenyl), fluorobenzyl (especially p-fluorobenzyl) and Fluorophenyl.

 Some particularly preferred specific compounds of the invention are listed below,

 The compound of the present invention may be in the form of a pharmaceutically acceptable acid addition salt thereof, which may be combined with a mineral acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., and an organic acid such as acetic acid, trifluoroacetic acid, Propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, a salt formed of toluenesulfonic acid, salicylic acid or the like. Method for preparing compounds of the invention

 The preparation method of the compound of the formula (I) provided by the invention comprises the following steps:

(a) a protecting group RJ of an amino group in the compound of the formula (II), which is selected from the group consisting of <^~0: ₆ alkane optionally substituted by a C ₆ -C _I() aryl group An oxycarbonyl group and a c,~c ₆ alkyl acyl group optionally substituted by a halogen, particularly a tert-butoxycarbonyl group, a trifluoroacetyl group or a benzyloxycarbonyl group;

(b) condensing the product of step (a) with a compound of formula (III) in the presence of a condensing reagent to provide a compound of formula (I),

 (I)

Ze i. 2001， 42， 6808-6809。 式 （III) 的化合物的合成可参 考文献： Dawei Ma, Weiqing Xie, Bin Zou, Qiong Lei and Duanqing Pei Tetrahedron Lett. 2004, 45, 8103-8105。 Wherein, the definitions of R ₂ , R 4 and U are the same as defined in the above formula (I). The synthesis of compounds of the formula (Π) can be found in the literature: Qingping Tian, Naresh K. Nayyar, Srinivasan Babu, Lijian Chen, Junhua Tao, Steven Lee, Anthony Tibbetts, Terence Moran, Jason Liou, Ming Guo and Timothy P. Kennedy

Ze i. 2001, 42, 6808-6809. The synthesis of compounds of formula (III) can be found in the literature: Dawei Ma, Weiqing Xie, Bin Zou, Qiong Lei and Duanqing Pei Tetrahedron Lett. 2004, 45, 8103-8105.

In a preferred embodiment of the present invention, the compound of the formula (Π) is reacted with an acid (for example, a mixture of dichloromethane and trifluoroacetic acid in a volume ratio of 1 to 4:1) in an organic solvent at room temperature for 1 to 4 Hour solvent; dissolve the product with a compound of formula (III) in an aprotic solvent (eg CH ₂ C1 ₂ , THF, CHCI3), add a condensation reagent (eg HATU, HBTU or EDCI), then add an organic base ( The reaction is carried out at room temperature for 8 to 24 hours, such as ipr ₂ Net or NEt ₃ ), to give a compound of the formula (I).

叔丁氧羰基、 异噁唑基羰 It is also possible to replace a group of the compound of the formula (I) with the intended group IV by derivatization, thereby obtaining a series of derivatives of the compound of the formula (I), wherein the group consisting of the following groups is selected group: halogen optionally substituted C ₃ ~ C ₆ cycloalkyl alkyl with a carbonyl group (^ - (^ alkyl with a carbonyl group, optionally <embankment ₆ ~ 0 ^ oxycarbonyl group is C ₆ ~C _{1 ()} of the substituted aryl group, C ₆ ~ a C _{1 ()} arylcarbonyl group and a C ₅ ～C _1Q heteroarylcarbonyl group optionally substituted by (^~0: ₄ fluorenyl); the derivatization described includes the following steps: (c) the obtained formula (I) Deprotection of the compound; (d) condensation of the product of step (c) with the carboxylic acid IV-OH in the presence of a condensing reagent to give a derivatized compound of formula (I) wherein the group is replaced by a group I; wherein 'a group selected from the group consisting of: optionally substituted by halogen (^ - (^ alkylcarbonyl, C ₃ ~C ₆ cycloalkyl alkyl with a carbonyl group, optionally substituted C ₆ ~C _{1 ()} aryl substituted (^~( ₆垸 oxycarbonyl, 0 ₆ 〜. arylcarbonyl and optionally substituted by C, ~C ₄ alkyl C ₅ ～d. Heteroarylcarbonyl, such as trifluoromethyl) Carbonyl, benzyloxycarbonyl,

Tert-butoxycarbonyl, isoxazolylcarbonyl

Base, furanylcarbonyl,

In some preferred embodiments of the invention, the compound of formula (I) to be derivatized is reacted in an organic solvent with an acid (for example, a mixture of dichloromethane and trifluoroacetic acid in a volume ratio of from 1 to 4:1) at room temperature. 1~4 hours; remove the solvent, dissolve the product with the carboxylic acid R-OH in an aprotic solvent (such as CH ₂ C1 ₂ , THF, CHC1 ₃ ), add a condensation reagent (such as HATU, HBTU or EDCI), then add An organic base (e.g., ipr ₂ Net or NEt ₃ ) is allowed to react at room temperature for 8 to 24 hours to provide a series of derivatives of the compound of formula (I).

Wherein, the acid described above is preferably trifluoroacetic acid or hydrochloric acid; a preferred organic solvent is selected from the group consisting of CH ₂ C1 ₂ , tetrahydrofuran, CHC1 ₃ , N,N-dimethylformamide and dioxane Ring; preferred aprotic solvent is selected from the group consisting of CH ₂ C1 ₂ , tetrahydrofuran, CHC1 ₃ , hydrazine, hydrazine-dimethylformamide, dimethyl sulfoxide and benzene; preferred condensation reagents are selected from the following members Groups constituted: HATU, HBTU and EDCI; preferred organic bases are selected from the group consisting of diisopropylethylamine and triethylamine. Pharmaceutical compositions and preparations

The pharmaceutical compositions of the present invention comprise a small molecule inhibitor of the invention together with one or more pharmaceutically acceptable carriers. The compositions may take a wide variety of formulations, and the pharmaceutically acceptable carrier may be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules and capsules. The solid carrier can be one or more substances which may also act as a diluent, a flavoring agent, a solubilizer, a lubricant, a suspending agent, a binder or a tablet disintegrating agent; it may also be an encapsulating material. The preparation of various preparations can be carried out in accordance with a conventional method in the field of pharmacy.

 Preferably, the pharmaceutical preparation is in unit dosage form, such as a packaged preparation, the package containing discrete amounts of preparation such as packaged tablets, capsules and vials or powders in ampoules.

 The dosage can vary depending on the needs of the patient, the severity of the disease, and the particular compound employed. For convenience, the total daily dose can be divided into several doses throughout the day. The skilled person in the medical field is able to determine the appropriate dosage depending on the specific situation. In order to explain the present invention in more detail, specific embodiments of the present invention will be described below in conjunction with the accompanying drawings. In describing the embodiments, well-known experimental methods, apparatus, reagents, materials, and the like have not been described in detail to avoid the disadvantages of the present invention. Example 1

溶解在 2 ml CH₂C1₂中， 加入 0.5 mlWill be 42 mg

Dissolved in 2 ml CH ₂ C1 ₂ , added 0.5 ml

TFA, reacted at room temperature for 1 hour, and the solvent was evaporated. Dissolve the obtained de-Boc substrate

Ipr ₂ NEt, then add 63 mg HATU. After reacting for 12 hours at room temperature, it was washed with 1 M HCK saturated NaHCO^ solution, saturated brine and dried over Na ₂ SO ₄ . filter, The solvent was evaporated under reduced pressure and flash column chromatography afforded 52 mg.

 The spectrum data is as follows:

Ή NMR: δ (500 MHz, CDC1 ₃ ) 0.76 (d, 3H, J = 6.9 Hz), 0.98 (d, 3H _; J = 5.7 Hz), 1.30 (t, 3H, J = 7.3 Hz), 1.42 (s , 9H), 1.74-1.94 (m, 4H): 2.10-2.40 (m, 2H), 2.49-2.52 (m, IH), 2.67-2.71 (m, IH), 2.85-3.00 (m _; IH), 3.05 -3.17 (m, 1H), 3.23-3.39 (m, 3H), 4.18 (q, 2H, J = 7.1 Hz): 4.41-4.51 (m, 1H), 4.58-4.65 (m, IH), 5.03-5.10 (m, IH), 5.49 (t, IH, J = 14.7 Hz), 5.98 (dd, 1H, J = 15.8 Hz, 4.4 Hz), 6.70-7.00 (m, 3H) _; 7.12-7.16 (m, 3H) ;

ESI-MS: [M + H +] 590.2, HRMS Found _{m / z 612.3062: C 31 H} 44 N 3 0 7 Fna calcd 612.3065;

[ ] _D 24.0 (c 0.69, CHC1 ₃ ). Example 1

溶解在 2 ml CH₂C1₂中， 加入 0.5 ml TFA， 在室温下反应 1小时， 抽干溶剂。 将得到的脱 Boc Will be 45 mg

It was dissolved in 2 ml of CH ₂ C1 ₂ , added with 0.5 ml of TFA, and reacted at room temperature for 1 hour, and the solvent was evaporated. Will get off Boc

，再加入 48 μΐ ipr₂NEt， 然后加入 44 mg HATU。在室温下反应 12小时， 依次用 1 M HC 饱和 NaHCO^ 溶液、 饱和食盐水洗涤， 用 Na₂S0₄干燥。 过 减压蒸去溶剂， 快速柱层析， 得到 34 mg 产物 The substrate is dissolved in 2 ml CH ₂ C1 ₂ and 12 mg is added.

Then add 48 μΐ ipr ₂ NEt and then add 44 mg HATU. The reaction at room temperature for 12 hours, washed with 1 M HC saturated NaHCO ^ solution, saturated brine, dried with Na ₂ S0 _4. Over The solvent was evaporated under reduced pressure and flash column chromatography afforded

 The spectrum data is as follows:

Ή NMR: δ (300 MHz, CDC1 ₃ ) 0.85 (d, 3H, J = 6.6 Hz), 1.03 (d, 3H, J = 6.9 Hz), 1.30 (t, 3H, J = 7.2 Hz), 1.52-1.61 (m, IH), 1.71-1.90 (m, 2H), 2.26-2.42 (m, 2H), 2.48 (s, 3H), 2.53-2.73 (m, 3H), 2.84-2.98 (m, 2H), 3.13 -3.23 (m, IH), 3.30-3.42 (m, 2H), 4.18 (q, 2H, J = 7.2 Hz), 4.41-4.56 (m, IH), 4.66-4.76 (m, IH), 5.50 (d , 1H, J = 15.6 Hz), 5.91 (s, IH), 6.39 (s, IH), 6.63 (dd, 1H, J = 15.3 Hz, 4.8 Hz), 6.94-7.03 (m, 2H), 7.11-7.35 (m, 4H) _;

ESI-MS: [M + H +] 599.3, [M + Na +] 621.3, HRMS Found _{m / z 621.2717, C 31 H} 44 N 3 0 7 FNa calcd 621.2695;

[ ] _D 32.8 (c 0.51, CHCl ₃ ).

Example 1 and Example 2 can be briefly summarized by the following reaction formula:

采用与上述方法类似的方法制备以下化合物:

The following compounds were prepared in a similar manner to the above procedure:

NMR NMR (300MHz, CDC1 ₃ ) δ 8.13-8.05 (m, IH), 7.63-7.54 (m, 1H), 7.412 (m, 2H), 7.29 (m, 3H), 6.86-6.66 (m, 2H), 5.95-5.87 (m, IH), 5.67-4.82 (m, 2H), 4.74-4.50 (m, 1H), 4.21-4.02 (m, 3H), 3.34-3.21 (m, 2H), 2.71-2.31 (m , 2H), 2.14-1.91 (m, 3H), 1.84-1.68 (m, IH), 1.66-1.53 (m, 1H), 1.42 (s, 9H), 1.30-1.21 (m, 3H), 0.96-0.81 (m, 6H).

NMR NMR (300 MHz, CDC1 ₃ ) δ 8.25-8.04 (m, IH), 7.75-7.61 (m, IH) 7.47 (m, IH), 7.40 (m, 2H), 7.30 (m, 3H), 6.85- 6.67 (m, 1H), 6.42 (d, J = 8.7 Hz, IH), 5.97-5.80 (m, IH), 5.62 (m, IH), 4.77 (m, 1H), 4.57 (m, IH) 4.12 (m, 2H), 3.33 (m, 2H), 2.45 (s, 3H), 2.41 (m, IH), 2.24-2.01 (m, 4H) _: 1.98- 1.76 (m, IH), 1.71-1.44 (m, IH), 1.25 (m, 3H), 1.00-0.88 (m, 6H).

NMR NMR (300 MHz, CDC1 ₃ ) δ 8.30 (m, IH), 8.10 (m, IH), 7.52 (m, IH), 7.41-7.30 (m, 5H), 7.01 (m, IH), 6.85-6.65 (m, 1H), 5.97-5.78 (m, IH), 5.50 (m, IH), 4.54 (m, 1H), 4.38 (m, IH), 4.18 (m, 2H), 3.35-3.25 (m, 2H ), 2.47 (m, 1H), 2.22-2.09 (m, 2H), 1.87 (m, 2H), 1.43 (m, 1H), 1.26 (m, 4H), 0.96-0.85 (m, 8H), 0.74 ( m, 2H).

ESI-MS m / z 549 ( M + Na) +; HRMS found ^{mlz 549.2681 (M + Na) +} , C 28 H 38 N 4 0 6 Na calc 549.2684.

NMR NMR (300 MHz, CD ₃ OD) δ 7.88-7.80 (m, 3H), 7.56-7.41 (m _: 7H), 7.40-7.33 (m, 4H), 6.97-6.93 (m, 1H), 6.04 (m , 1H), 5.45 (m, 1H): 4.60 (m, IH), 4.41 (m, IH), 4.17 (m, 2H), 3.71 (m, 2H), 2.19 (m, 2H) _: 1.93 (m, 2H), 1.54 (m, 2H), 1.24 (m, 4H), 1.03 (m, 6H) _;

ESI-MS m / z 585 ( M + Na) +; HRMS found ^{mlz 563.2862 (M + H) +} : C 31 H 39 N 4 0 6 calcd 549.2684. Example 3

 In describing the present embodiment, reference is made to the following reaction formula (where one or two Arabic numerals below some of the compounds are numbered):

92%

N1 Preparation of compound 9

， 0 4 ml二异丙基乙胺， 162 mg HOBt,最后加入 247 mg DCC, 在室温下搅拌过夜。 将反应体系过滤， 加入 10 ml二氯甲烷， 依次用 1 M HC1、 饱和碳酸氢钠水溶液、 饱和氯化钠水溶液洗涤， 用 无水硫酸钠干燥， 过滤， 蒸去溶剂， 快速柱层析， 得到 387 mg化合 物 9， 收率 92%。 321 mg (1 mmol) of Compound 8 was dissolved in 2 ml of dichloromethane, and 1 ml of trifluoroacetic acid was added thereto, and the mixture was stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure and the oil pump was drained. The obtained product was dissolved in 4 ml of dichloromethane, and 260 mg of the compound was added in sequence.

, 0 4 ml of diisopropylethylamine, 162 mg of HOBt, and finally 247 mg of DCC, and stirred at room temperature overnight. The reaction system was filtered, and 10 ml of methylene chloride was added, and the mixture was washed with 1 M EtOAc, EtOAc. 387 mg of compound 9, yield 92%.

 The spectrum data is as follows:

Ή NMR: δ (300 MHz, CDC1 ₃ ) 0.90-0.95 (m, 12H), 1.44 (s, 9H), 1.45-1.68 (m, 3H), 2.03 -2.15 (m, 1H), 3.88 (dd, IH , J = 6.6 Hz, 9.3 Hz), 4.64-4.72 (m, IH), 5.06 (d, IH, J = 9.9 Hz), 5.16 (d, 2H, J = 3.3 Hz), 6.24 (d, IH, J = 10.5 Hz), 7.29-7.30 (m, 5H). Preparation of Compound 10

331 mg of Compound 9 was dissolved in 4 ml of dichloromethane, and 1 ml of trifluoroacetic acid was added thereto, and the mixture was stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure and the oil pump was drained. The obtained product was dissolved in 4 ml of dichloromethane, and 19 mg of compound ^B ° ^{cH N} "^ ^C0 2H, _0.3 ml of diisopropylethylamine, 128 mg of HOBt, and finally 195. mg of DCC were added. After stirring at room temperature, the reaction system was filtered, and 10 ml of dichloromethane was added, and then washed with 1 M HCl, saturated aqueous sodium hydrogencarbonate and saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, evaporated Solvent, flash column chromatography gave 357 mg of compound 10, yield 92%.

 The spectrum data is as follows:

Ή NMR: δ (300 MHz, CDC1 ₃ ) 0.89-0.94 (m, 12H), 1.35 (d, 3H, J = 7.2 Hz), 1.44 (s, 9H), 1.56-1.69 (m, 3H), 2.13 - 2.24 (m, IH), 4.09-4.19 (m, 1H), 4.24 (dd, 1H, J = 6.6 Hz, 8.7 Hz), 4.92-5.03 (m, 1H), 4.95 (d, 2H _: J = 5.4 Hz), 6.35-6.44 (m, 1H), 6.69-6.78 (m, 2H), 7.31-7.42 (m, 5H). Preparation of Compound 11

 310 mg of compound 10 was dissolved in 5 ml of methanol, 62 mg of 20% palladium on carbon was added, hydrogenation was carried out at atmospheric pressure for 3 hours, palladium carbon was filtered off, the solvent was evaporated, and the column was purified by column chromatography to give 258 mg of compound 11. 100%.

 The spectrum data is as follows:

Ή NMR: δ (300 MHz, CDC1 ₃ ) 0.91-0.96 (m, 12H), 1.34 (d, 3H, J = 6.9 Hz), 1.44 (s, 9H), 1.60-1.75 (m, 3H), 2.08 - 2.20 (m, IH), 4.18-4.28 (m, 1H), 4.33 (t, 1H, J = 9.0 Hz), 4.51-4.61 (m, IH), 5.27-5.35 (m, IH), 7.08-7.17 ( m, 1H), 7.27-7.33 (m, 1H). Preparation of compound N2

溶解在 2 ml CH₂C1₂中， 加入 0.5 mlWill be 50 mg

Dissolved in 2 ml CH ₂ C1 ₂ , added 0.5 ml

TFA, reacted at room temperature for 1 hour, and the solvent was drained. The obtained Boc-depleted substrate was dissolved in 2 ml of CH ₂ C1 ₂ , 62 mg of compound 11 was added, and 97 μM of 'P^NEt was added, followed by 75 mg of HATU. The reaction at room temperature for 12 hours, washed with 1 M HC1, saturated aqueous NaHC0 _3, and saturated brine, dried over Na ₂ S0 _4, filtered, and the solvent was evaporated under reduced pressure, flash column chromatography to give 76 mg product N2, close The rate is 81%.

 The spectrum data is as follows:

'H NMR (300MHz, CDC1 ₃ ) δ 0.89-1.02(m, 12H) 1.25-1.31 (m, 3H) 1.36-1.40 (m, 2H) 1.42 and 1.45 (2 xs, 9H) 1.51 -1.68 (m, 3H) ) 1.69-1.97 (m, 2H) 2.04-2.15 (m, IH) 2.19-2.57 (m, 4H) 3.22-3.40 (m, 2H) 4.11-4.35 (m, 3H) 4.47-4.61 (m, IH) 4.67 -4.78 (m, 2H) 5.03-5.12 (m, IH) 5.24 (d, IH, J=3.3) 5.87-6.00 (m, 2H), 6.64-6.73 (m, IH), 6.83-7.00 (m, 2H) ); ESI-MS m/z 610.4 (M+H)+. Preparation of Compound Nl

41 mg of the compound N2 was dissolved in 2 ml of CH ₂ C1 ₂ , 0.5 ml of TFA was added, and the mixture was reacted at room temperature for 1 hour, and the solvent was evaporated. Dissolve the resulting deBoc substrate in 2 ml THF

， 再加入 42 μΓΡΓ₂ΝΕί， 然后加入 33 mg HATU。 在室温下反应 12小时， 依次用 1 M HC1、 饱和 NaHCO^_ 液、 饱和食盐水洗涤， 用 Na₂S0₄干燥。 过滤， 减压蒸去溶剂， 快速 柱层析， 得到 34 mg产物 N1 , 收率 82%。 Medium, add 10 mg

, add 42 μΓΡΓ ₂ ΝΕί, then add 33 mg HATU. After reacting for 12 hours at room temperature, it was washed with 1 M HCl, saturated NaHCO^ solution, brine, and dried over Na ₂ SO ₄ . Filtration, evaporation of the solvent under reduced pressure and flash column chromatography afforded,

 The spectrum data is as follows:

Ή NMR: δ (300 MHz, CDC1 ₃ ) 0.89 (s, 12H), 1.25-1.29 (m, 3H), 1.44 (d, 3H, J = 7.4 Hz), 1.50-1.92 (m, 5H), 2.04- 2.17 (m, 2H), 2.20 -2.41 (m, 3H), 2.47 (s, 3H), 3.20-3.41 (m, 2H), 4.18 (q, 2H, J = 7.2 Hz), 4.32-4.43 (m, 1H), 4.57-4.79 (m, 2H), 4.85-4.97 (m, 1H), 5.93-6.00 (m, 1H), 6.46 (s, 1H), 6.82-6.92 (m, 1H), 7.47-7.93 ( m, 3H);

 ESI-MS m/z 619.3 (M+H)+. The following compounds were prepared in a similar manner to the above procedure:

NMR NMR (300 MHz, CDC1 ₃ ) δ 0.75-0.95 (m, 12H), 1.42 (d, J = 7.2 Hz, 3H), 1.48-1.85 (m, 5H), 1.95-2.40 (m, 7H), 2.44 (s, 3H), 3.20-3.37 (m, 2H), 4.26-4.40 (m, 1H), 4.53-4.77 (m, 2H), 4.82-4.92 (m, 1H), 5.16 (s, 2H), 6.02 (dd, J = 5.7 Hz, 15.9 Hz, 1H), 6.44 (s, 1H), 6.67 and 6.73 (2 x brs, 1H), 6.87-7.95 (m, 1H), 7.28-7.43 (m, 5H), 7.45-7.67 (m, 1H), 7.34-7.84 (m 1H); ESI-MS w/ 681 (M+H) ten.

NMR NMR (300 MHz, CDC1 ₃ ) δ 0.86-1.02 (m, 12H), 1.37 (dd, J = 3.9 Hz, 6.9 Hz, 3H), 1.42, 1.43 and 1.44 (3 xs, 9H), 1.50-1.95 ( m, 6H), 2.22—2.55(m, 4H), 3.22-3.48 (m, 2H), 4.05-4.13 (m, 2H), 4.47-4.78 (m, 2H), 5.02-5.10 (m, 1H), 5.14-5.21 (m, 2H), 5.83-5.88 (m, 1H), 5.95-6.12 (m, 1H), 6.61-6.68 (m, 1H), 6.88-6.99 (m, 1H), 7.30-7.40 (m , 6H);

ESI-MS m/z 672 (M+H) ⁺ . Example 4

Crystal preparation, data collection and structural analysis of a complex of SARS-CoV M ^pro with small molecule inhibitors

I. Materials and methods

1. Crystal preparation of a complex of small molecule compound N1 with 1 8 8 11⁄2-( 0¥ ]\ ^°

 Will SARS-CoV! ^^ is further isolated, purified and crystallized after expression in E. coli strain BL21 (DE3) (see: Yang H et al. 2003. The Crystal Structures of SARS Virus Main Protease Mpro and Its Complex with an Inhibitor. PNAS, 100 (23) ): 13190-13195). After the N1 was prepared into a 10 mM solution with a pool of 7.5% PEG 6000, 6% DMSO, 0.1 M MES (pH 6.0), an equal volume was added to the hanging suspension of the crystal growth, and 291 K was allowed to stand for 2 days.

2. 8 8 1 ^ 0 ^ ] \ ^ ° and small molecular compound N1 complex data collection and structural analysis

 (1) Data collection and data processing

Completed with Rigaku αιΛ "α rotating target X-ray diffractometer, system parameters for data collection The voltage is 40 KV, the current is 20 mA, the wavelength is 1.5418A, and the temperature is 100K. The crystals were immersed in an antifreeze of 30% PEG 400 0.1 M MES (pH 6.0) and rapidly cooled to 100 K with a liquid nitrogen stream. The diffraction resolution of the crystal is 1.88 Å. All final diffraction data was processed to HKD2000 to 2.0 Α

(2) Structure analysis

Composite structure analysis by using the parent structure of the SARS-CoV M ^pTO (PDB code: 1UJ1) as a starting search model, structural analysis by molecular replacement. Using the CNS program to do the rotation function and the translation function search, a clear solution of a symmetric two-body can be obtained. The small molecule model was then constructed using the difference maps of IFo-Fc and Fo-Fc in program 0. After correction, R _k 20.0, R _free 23.8

 The method of crystal preparation, data collection and structural analysis of the composite of SARS-CoV ^^° and N2 N3 and N4 is basically the same as that of N1, and will not be described here. Second, structural analysis

Structural Analysis of the Complex of 1 5 8 1^-0^ 1^" ⁾ with Small Molecule Compound N1

8 八 1^-0^ ^ ¹ "° is a homodimeric structure containing two monomers A and B in one asymmetric unit, since the combination of the small molecule compound Nl and AB is basically similar. Now only analyze the combination mode of N1 and A.

In the 2Fo-Fc difference map (countered at 1σ) of monomer A, it was clearly observed that N1 binds to the substrate binding pocket of the enzyme. Among them, C 3 in N1 forms a covalent bond with S _{7 of} A145-Cys having a bond length of 1.8A. The carbonyl oxygen in the N1 ester group forms a hydrogen bond with the amino group on the A145-Cys main carbon chain; the ethyl group in the ester forms a hydrophobic interaction with the side chains of A27-Len His-A41 and Thr-A25. At the P1 site, the oxygen in the five-membered ring of the lactam forms a 2.6A hydrogen bond with NE2 of His-163. A water molecule near the lactam ring can form a 2.6A 3.2A, 2.6 person and 2.7 human hydrogen bond with NE2 of N His-172 on the lactam ring of N1 and carbonyl oxygen of 140-Phe and Ser-Bl, respectively. , allowing the lactam ring to be firmly bonded in the S1 pocket. P1 due to the major protease of the entire coronavirus family The site is very conserved and always appears in the form of Q, therefore, occupying the corresponding substrate binding pocket S1 is a key to inhibiting protease activity. For the P2 site of N1, the small molecule Leu side chain is easily inserted into the side chain of His-41, Met-49 and Phe-181, and the alkyl moiety of the side chain of Gln-189 and Asp-187. In the hydrophobic pocket. The carbonyl oxygen in His-164 forms a hydrogen bond of 2.9A with N in the peptide bond near the ester group side in N1, and the carbonyl oxygen of Val in N1 forms a 2.9A hydrogen bond with N in GKI-A166, Val N in the peptide bond forms a hydrogen bond of 3.0A with the G1U-A166 carbonyl oxygen, and N in the Ala peptide bond in N1 forms a hydrogen bond of 3.2A with the carbonyl oxygen of Thr-190. The side chain of Ala is inserted into a hydrophobic pocket consisting of Phe-185, Glu-192, Leu-167, Met-165 side chains. The heterocyclic ring at the end of N1 has a hydrophobic interaction with the five-membered ring of Pro-168. All of these covalent bonds, hydrogen bonds, and hydrophobic interactions allow the inhibitor compound N1 to bind tightly to the substrate active pocket of the enzyme, thereby causing inactivation of the enzyme.

2. The combination of N2 and N3 is basically similar to that of N1, and will not be described here. Example 5

Small molecule compounds Nl, N2, N3 and N4 inhibit the main protease from various coronaviruses

I. Materials and methods

1. Determination of inhibitory activities of N1, N2, N3 and N4 on SARS-CoV M ^pro

In a buffer (20 mM Tris-HCl H 7.0, 1 mM DTT), add SARS-CoV M ^pro (final concentration 1 μΜ), compound Nl (final concentration 100 μΜ), 298 Κ for 10 minutes, then quickly add fluorescence Label the substrate (MCA-AVLQ SGFRL (DNP) L-NH ₂ , 5 μΜ). The excitation and emission wavelengths were 330 η Μ and 395 nm, respectively, and the temperature was maintained at 298 K. Fluorescence readings were recorded every 2 seconds.

 The method for measuring the inhibitory activities of Ν2, Ν3 and Ν4 is basically the same as that of N1.

 Control: No inhibitor was added and the rest of the conditions were the same. The results are shown in Figure 3.

2. Determination of inhibitory activity of N1 against infectious gastrointestinal virus (TGEV) major protease In the buffer (20 mM Tris-HCl pH 7.0, ImMDTT), the protease (1 μM), compound Nl (100 μM), 298 Κ was added for 10 minutes, and then the fluorescently labeled substrate (MCA-AVLQSGFRL (DNP) L was quickly added. -NH ₂ , 5 μΜ). The excitation and emission wavelengths were 330 η Μ and 395 nm, respectively, and the temperature was maintained at 298 K. Fluorescence readings were recorded every 2 seconds. Then, the inhibitor concentration was changed, and the inhibitory activity was measured at 10 η Torr and 1 μ 分别, respectively. Control: No inhibitor was added and the rest of the conditions were the same. The results are shown in Figure 4.

3. Determination of the inhibitory activity of N1 on human coronavirus (HCoV) 229E main protease

In the buffer (20 mM Tris-HCl pH 7.0, 1 mMDTT), the protease (0.1 μM), compound Nl (100 μM), 298 Κ was placed for 10 minutes, and then the fluorescently labeled substrate (MCA-AVLQSGFRL (DNP) was rapidly added. ) L-NH ₂ , 5 μΜ). The excitation and emission wavelengths were 330 η Μ and 395 nm, respectively, and the temperature was maintained at 298 K. Fluorescence readings were recorded every 2 seconds. Control: No inhibitor was added and the rest of the conditions were the same. The results are shown in Figure 5.

4. Determination of the inhibitory activity of N1 against the main protease of feline infectious peritonitis virus (FIPV) The same method as in 3, the results are shown in Fig. 6.

5. Determination of the inhibitory activity of N1 against the main protease of avian infectious bronchitis virus (AIBV)

In the buffer (20 mMTris-HCl pH 7.0, 1 mMDTT), the protease (0.1 μΜ), compound Nl (100 μΜ), 298 Κ was added for 10 minutes, and then the fluorescently labeled substrate (MCA-AVLQSGFRL (DNP) was quickly added. L-NH ₂ , 5 μΜ). The excitation and emission wavelengths were 330 η Μ and 395 nm, respectively, and the temperature was maintained at 298 K. Fluorescence readings were recorded every 2 seconds. Control: No inhibitor was added and the rest of the conditions were the same. The results are shown in Fig. 7.

Expression and purification of FIPV, HCoV, TGEV References: Conservation of substrate specificities among coronavirus main proteases. J. Gen. Virol. 2002; 83 (Pt 3): 595-9.

Second, the results and analysis

1. Inhibitory activity of Nl, N2, N3 and N4 on SARS-CoV M ^pf °

As can be seen from Fig. 3, Nl, N2, N3 and N4 have inhibitory activities against SARS-CoV M ^pro , and N1 has the strongest inhibitory activity.

 2. Inhibitory activity of N1 against other major proteases of coronavirus

 From the inhibition activity of N1 against other coronavirus main proteases (ie, Fig. 4 to Fig. 7), N1 has an inhibitory activity against the main proteases of TGEV, HCoV, FIPV and AIBV, and the most inhibitory activity against the main protease of TGEV. Strong, under the condition of ΙΟ ιιΜ, N1 still has inhibitory activity against TGEV main protease.

 Since TGEV, HCoV, and FIPV belong to the type I (serotype) of the Coronavirus family, AIBV belongs to type III, and SARS-CoV belongs to type IV. Therefore, it can be inferred that N1 has inhibitory activity against the major protease of the entire coronavirus family. References referred to herein, including patent documents, academic papers, publications, etc., are hereby incorporated by reference in their entirety.

 It should be noted that the various experimental operations involved in the present invention are conventional techniques in the art. If not specifically described herein, those skilled in the art can refer to various commonly used reference books and technologies before the filing date of the present application. Documents or related manuals, manuals, etc. are implemented.

 Among the various experimental products (including but not limited to: chemical reagents, biological products, cells, organisms, instruments, etc.) involved in this article, for those special or difficult to obtain, the manufacturer has been identified in the text. Reference documents or detailed preparation methods; unless otherwise specified, are conventional laboratory articles, which can be conveniently obtained by various means (for example, purchase, self-preparation, etc.) before the filing date of the present application.

 It is to be understood that various changes and modifications can be made in the form and details without departing from the spirit and scope of the invention, which are considered to fall within the scope of the present invention. in.

Claims

Rights request 1. A compound of the following formula (I) and a pharmaceutically acceptable acid addition salt or solvate thereof u is

Or CH: R, is selected from the group consisting of group consisting of: ~C optionally substituted by halogen (^ alkyl with _1-6 carbonyl, C ₃ ~C ₆ cycloalkyl alkyl with a carbonyl group, optionally substituted C ₆ ~C _1Q aryl group substituted ₆ Alkoxycarbonyl, C ₆ ～d. arylcarbonyl and optionally substituted by (^~(: ₄ fluorenyl). Heteroarylcarbonyl; R ₂ groups selected from the group consisting of: wherein aryl is optionally substituted by halogen, optionally substituted C ₆ ~ C ₁₀ aryl group substituted ~C ₆ alkyl with C ₆ ~C _IQ and an aryl group; R _{3 is} selected from the group consisting of: wherein the aryl group is optionally substituted by halogen, optionally substituted by a C ₆ -C _1Q aryl group (^~(: ₆ alkyl, optionally by (^~(: ₄₎ Alkyl-substituted C ₆ -C ₁₀ aryl; R4 is selected from the group group consisting of: wherein aryl is optionally substituted with halogen, optionally substituted C _{6 ~C 1Q} aryl ~C ₆ alkyl, optionally substituted (~ ^ _<4 alkyl with C _{6 to} C ₁₀ aryl; R _{5 is} selected from the group consisting of a group in which an aryl group is optionally substituted by a halogen, optionally substituted with a C ₆ -C _1Q aryl group ( ₆ alkyl group, optionally by (^~(: ₄垸) a C ₆ ～C ₁₍₎ aryl group substituted with a base and/or a halogen. 2. A compound of claim 1 or a pharmaceutically acceptable acid addition salt or solvate thereof, wherein U is

And X is NH. 3. A compound of claim 1 or a pharmaceutically acceptable acid addition salt or solvate thereof, Among them, U

, and X is NH. 4. The compound of claim 1 or a pharmaceutically acceptable acid addition salt or solvate thereof, Where U is

, and X is CH ₂ . 5. A compound according to any one of claims 1 to 4, or a pharmaceutically acceptable acid addition salt or solvate thereof, wherein: a group selected from the group consisting of trifluoromethylcarbonyl, benzyloxycarbonyl,

Tert-butoxycarbonyl, isoxazolylcarbonyl, furylcarbonyl,

 Is methyl, isopropyl, phenyl, benzyl, fluorobenzyl;  Selected from the group consisting of methyl, isopropyl, benzyl, fluorobenzyl (especially p-fluorobenzyl), phenyl and methylphenyl (especially p-methylphenyl);  R4 is selected from the group consisting of ethyl, phenyl, benzyl, methylphenyl (especially p-methylphenyl) and fluorobenzyl; R ₅ is isobutyl, phenyl, benzyl, methylphenyl (especially p-methylphenyl), fluorobenzyl (particularly p-fluorobenzyl) and fluorophenyl. 6. A compound according to claim 5, or a pharmaceutically acceptable acid addition salt or solvate thereof, wherein the compound is selected from the group consisting of:

 Wherein, Boc is a tert-butoxycarbonyl group, Ph is a phenyl group, Et is an ethyl group, and Bn is a benzyl group. The compound according to claim 6, wherein the structural formula of the compound is:

 8. A method of preparing a compound according to claim 1, comprising the steps of: (a) removing a protecting group of an amino group in the compound of the formula (II), wherein a group consisting of a group consisting of a < ₆ ~ ₆ alkoxycarbonyl group optionally substituted by a C ₆ -C _1Q aryl group and optionally a ~C ₆ decanoyl group substituted by a halogen;  (b) condensing the product of step (a) with a compound of formula (III) in the presence of a condensing reagent to provide a compound of formula (I),

 (II) (III)

 (I) Wherein R, R ₂ , R 4 and U have the same definitions as in claim 1. 9. The method of claim 8, wherein is a tert-butoxycarbonyl group, a trifluoroacetyl group or a benzyloxycarbonyl group. 10. The method of claim 8 comprising the steps of: (a) reacting a compound of formula (II) with an acid at room temperature in an organic solvent 1 to 4 After hours, remove the protecting group of the amino group and remove the solvent;  (b) The product of the step (a) and the compound of the formula (III) are dissolved in an aprotic solvent, and a condensation reagent and an organic base are added thereto, and the mixture is allowed to react at room temperature for 8 to 24 hours to obtain a compound of the formula (I). 11. The method of any of claims 8-10, further comprising the step of further derivatizing the resulting compound of formula (I) to replace the group therein with the desired group R, ', wherein 'a group selected from the group consisting of: optionally halogen-substituted alkyl with a carbonyl group, C ₃ ~C ₆ cycloalkyl alkyl with a carbonyl group, an optionally substituted C _{6 ~C 1 ()} aryl ~C ₆ oxygen embankment a carbonyl group, a C ₆ ～C _1Q arylcarbonyl group, and a C ₅ ～ heteroaryl carbonyl group optionally substituted by C, —C ₄ alkyl; — the derivatization described includes the following steps:  (c) in the compound of formula (I) obtained in the process of any one of claims 8-10! ^ remove;  (d) condensing the product of step (c) with a carboxylic acid R -OH in the presence of a condensing reagent to give a derivatized compound of formula (I) wherein the group is replaced by a R, ' group. 12. The method of claim 10, wherein  The acid is trifluoroacetic acid or hydrochloric acid; The organic solvent is selected from the group consisting of CH ₂ C1 ₂ , tetrahydrofuran, CHC1 ₃ , hydrazine, hydrazine-dimethylformamide and dioxane; The aprotic solvent is selected from the group consisting of C3⁄4C1 ₂ , tetrahydrofuran, CHC1 ₃ , N,N-dimethylformamide, dimethyl sulfoxide and benzene;  The condensation reagent is selected from the group consisting of: HATU, HBTU and EDCI; The organic base is selected from the group consisting of diisopropylethylamine and triethylamine. 13. A pharmaceutical composition comprising a compound of any one of claims 1-7, or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carriers. 14. Use of a compound according to any one of claims 1-7, or a pharmaceutically acceptable acid addition salt or solvate thereof, for the manufacture of a medicament for the treatment or prevention of a coronavirus infection. The use according to claim 14, wherein the coronavirus is SARS coronavirus, porcine transmissible gastroenteritis virus, human coronavirus, feline infectious peritonitis virus or avian infectious bronchitis virus.