RING-MODIFIED PROLINE SHORT PEPTIDE COMPOUND AND USE THEREOF

Abstract

Disclosed are a ring-modified proline short peptide compound and the use thereof, and specifically disclosed is a compound represented by formula (X) or a pharmaceutically acceptable salt thereof.

##STR00001##

Claims

1. A compound represented by formula (X) or a pharmaceutically acceptable salt thereof, ##STR00552## wherein, G is selected from ##STR00553## ##STR00554## ring A is selected from C.sub.3-10 cycloalkyl, 3- to 10-membered heterocycloalkyl, C.sub.6-10 aryl and 5- to 10-membered heteroaryl; R.sub.1 is each independently selected from halogen, OR.sub.11, CN, CH.sub.3S(O).sub.m-, -NH(R.sub.12), C.sub.1-3 alkyl and C.sub.1-3 haloalkyl; R.sub.11 is selected from H, C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, CH.sub.3(OCH.sub.2CH.sub.2).sub.p- and H(OCH.sub.2CH.sub.2).sub.q-; R.sub.12 is selected from C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, CH.sub.3CO— and CH.sub.3SO.sub.2—; m is selected from 0, 1 and 2; p and q are selected from 1, 2, 3, 4, 5 and 6; n is selected from 0, 1, 2, 3 and 4; X is selected from -CH(R.sub.3)-, —CH.sub.2CH.sub.2—, O, S, Se, SO.sub.2 and -N(R.sub.3)-, and the —CH.sub.2CH.sub.2— is optionally substituted by 1, 2, 3 or 4 R; R is each independently selected from halogen, OH, NH.sub.2, CN, C.sub.1-3 alkyl and C.sub.1-3 haloalkyl; R.sub.3 is each independently selected from H, C.sub.1-3 alkyl, C.sub.1-3 alkoxy, C.sub.1-3 haloalkyl, C.sub.1-3 haloalkoxy, C.sub.6-10 aryl and 5- to 10-membered heteroaryl, and the C.sub.6-10 aryl and 5- to 10-membered heteroaryl are optionally substituted by 1, 2 or 3 R.sub.31; R.sub.2 and R.sub.4 together with the atoms to which they are attached form C.sub.5-8 cycloalkyl, 5- to 6-membered heterocycloalkyl or 5- to 6-membered heterocycloalkenyl, and the C.sub.5-8 cycloalkyl, 5- to 6-membered heterocycloalkyl or 5- to 6-membered heterocycloalkenyl is optionally and independently substituted by 1 or 2 R.sub.a; R.sub.a is each independently selected from H, halogen, C.sub.1-3 alkyl, C.sub.1-3 alkoxy, C.sub.1-3 haloalkoxy, C.sub.3-6 cycloalkyl, C.sub.6-10 aryl and 5- to 10-membered heteroaryl, and the C.sub.3-6 cycloalkyl, C.sub.6-10 aryl and 5- to 10-membered heteroaryl are optionally substituted by 1, 2 or 3 R.sub.41; R.sub.21, R.sub.31 and R.sub.41 are each independently selected from halogen, OH, NH.sub.2, CN, C.sub.1-3 alkyl, C.sub.1-3 alkoxy, C.sub.1-3 haloalkyl and C.sub.1-3 haloalkoxy; R.sub.5 is selected from C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, C.sub.1-3 alkoxy, C.sub.1-3 haloalkoxy, -CH.sub.2-R.sub.6 and -CH.sub.2-O-R.sub.6; R.sub.6 is selected from phenyl, and the phenyl is optionally substituted by 1, 2 or 3 R.sub.61; R.sub.61 is selected from halogen, C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, C.sub.1-3 alkoxy and C.sub.1-3 haloalkoxy.

2. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from structures represented by formulas (X-1) and (X-2), ##STR00555## ##STR00556## wherein, R.sub.b is each independently selected from H, halogen, C.sub.1-3 alkyl, C.sub.1-3 alkoxy, C.sub.1-3 haloalkoxy and C.sub.3-6 cycloalkyl; or, two R.sub.b on adjacent carbon atoms or the same carbon atom together with the atoms to which they are attached form cyclopropyl; t is selected from 1 and 2.

3. The compound or the pharmaceutically acceptable salt thereof according to claim 2, wherein, R.sub.b is each independently selected from H, F, methyl, ethyl, isopropyl and cyclopropyl.

4. The compound or the pharmaceutically acceptable salt thereof according to claim 2, wherein, the structural moiety ##STR00557## is selected from ##STR00558## ##STR00559## ##STR00560## ##STR00561## ##STR00562## ##STR00563## ##STR00564## ##STR00565## ##STR00566## ##STR00567## ##STR00568## .

5. The compound or the pharmaceutically acceptable salt thereof according to claim 4, wherein, the structural moiety ##STR00569## is selected from ##STR00570## ##STR00571## ##STR00572## ##STR00573## ##STR00574## ##STR00575## ##STR00576## ##STR00577## ##STR00578## ##STR00579## ##STR00580## .

6. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.1 is each independently selected from halogen, C.sub.1-3 alkyl, C.sub.1-3 alkoxy, C.sub.1-3 haloalkyl and C.sub.1-3 haloalkoxy.

7. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.1 is each independently selected from F, Cl and methyl.

8. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, ring A is selected from C.sub.5-10 cycloalkyl and phenyl.

9. The compound or the pharmaceutically acceptable salt thereof according to claim 8, wherein, ring A is selected from cyclohexyl, spiro[3.3]heptyl, bicyclo[2.2.2]octyl, adamantyl and phenyl.

10. The compound or the pharmaceutically acceptable salt thereof according to claim 9, wherein, ring A is selected from ##STR00581## ##STR00582## ##STR00583## ##STR00584## ##STR00585## .

11. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, the structural moiety ##STR00586## is selected from ##STR00587## ##STR00588## ##STR00589## ##STR00590## ##STR00591## ##STR00592## ##STR00593## .

12. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein, R.sub.5 is selected from —CF.sub.3, —OCH.sub.3, ##STR00594## ##STR00595## .

13. A compound represented by the following formula or a pharmaceutically acceptable salt thereof, selected from ##STR00596## ##STR00597## ##STR00598## ##STR00599## ##STR00600## ##STR00601## ##STR00602## ##STR00603## ##STR00604## ##STR00605## ##STR00606## ##STR00607## ##STR00608## ##STR00609## ##STR00610## ##STR00611## ##STR00612## ##STR00613## ##STR00614## ##STR00615## ##STR00616## .

14. A pharmaceutical composition, comprising the compound or the pharmaceutically acceptable salt thereof according to claim 1; further, the pharmaceutical composition can also comprise a pharmaceutically acceptable excipient.

15. A method for the treatment of a disease related to 3CL protease in a subject in need thereof, comprising: administering the compound or the pharmaceutically acceptable salt thereof according to claim 1 to the subject.

16. The method according to claim 15, wherein the disease related to 3CL protease is coronavirus infection.

17. The method according to claim 16, wherein the coronavirus infection is infection with COVID-19.

18. A compound represented by formula (I′) or a pharmaceutically acceptable salt thereof, ##STR00617## wherein, the structural moiety ##STR00618## ##STR00619## ##STR00620## ##STR00621## ##STR00622## ##STR00623## and ##STR00624## are optionally and independently substituted by 1, 2 or 3 R .sub.a′; ring A′ is selected from C.sub.3-6 cycloalkyl, and the C.sub.3-6 cycloalkyl is optionally substituted by 1 or 2 R.sub.a′; ring B is selected from C.sub.4-8 cycloalkyl, C.sub.5-8 cycloalkenyl, 3- to 8-membered heterocycloalkyl and 5- to 8-membered heterocycloalkenyl, and the C.sub.4-8 cycloalkyl, C.sub.5-8 cycloalkenyl, 3- to 8-membered heterocycloalkyl and 5- to 8- membered heterocycloalkenyl are optionally substituted by 1 or 2 R.sub.a′; ring D is selected from C.sub.4-8 cycloalkyl, C.sub.5-8 cycloalkenyl and 5- to 8- membered heterocycloalkyl, and the C.sub.4-8 cycloalkyl is optionally substituted by 1 or 2 R.sub.a′; R.sub.a′ is each independently selected from F and methyl; the heterocycloalkyl contains 1, 2 or 3 heteroatoms or heteroatom groups independently selected from O, S, N and NH.

19. The compound or the pharmaceutically acceptable salt thereof according to claim 18, wherein, the structural moiety ##STR00625## is selected from ##STR00626## ##STR00627## ##STR00628## ##STR00629## or, the structural moiety ##STR00630## is selected from ##STR00631## ##STR00632## ##STR00633## ##STR00634## ##STR00635## ##STR00636## wherein W is selected from CH .sub.2, NH, N(CH.sub.3), O, S and SO.sub.2; or, the structural moiety ##STR00637## is selected from ##STR00638## ##STR00639## .

20. The compound or the pharmaceutically acceptable salt thereof according to claim 19, wherein, the structural moiety ##STR00640## is selected from ##STR00641## ##STR00642## ##STR00643## ##STR00644## ##STR00645## ##STR00646## ##STR00647## ##STR00648## ##STR00649## .

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0253] FIG. 1. The binding mode pattern of compound 15 and 6WTT protein;

[0254] FIG. 2. the binding mode pattern of compound 16 and 6WTT protein;

[0255] FIG. 3. the binding mode pattern of compound 17 and 6WTT protein;

[0256] FIG. 4. the binding mode pattern of compound 18 and 6WTT protein;

[0257] FIG. 5. the binding mode pattern of compound 19 and 6WTT protein;

[0258] FIG. 6. the binding mode pattern of compound 20 and 6WTT protein;

[0259] FIG. 7. the binding mode pattern of compound 21 and 6WTT protein;

[0260] FIG. 8. the binding mode pattern of compound 22 and 6WTT protein;

[0261] FIG. 9. the binding mode pattern of compound 23 and 6WTT protein;

[0262] FIG. 10. the binding mode pattern of compound 24 and 6WTT protein;

[0263] FIG. 11. the binding mode pattern of compound 25 and 6WTT protein;

[0264] FIG. 12. the binding mode pattern of compound 26 and 6WTT protein; FIG. 13. the binding mode pattern of compound 27 and 6WTT protein.

DETAILED DESCRIPTION OF THE INVENTION

[0265] The present disclosure is described in detail by the embodiments below, but it does not mean that there are any adverse restrictions on the present disclosure. The present disclosure has been described in detail herein, and its specific embodiments have also been disclosed; for one skilled in the art, it is obvious to make various modifications and improvements to the embodiments of the present disclosure without departing from the spirit and scope of the present disclosure.

Calculation Embodiment 1

Simulation of the Binding Mode of Compounds 15 to 27 to Protein:

##STR00517##

##STR00518##

##STR00519##

##STR00520##

##STR00521##

##STR00522##

##STR00523##

##STR00524##

##STR00525##

##STR00526##

##STR00527##

##STR00528##

##STR00529##

[0266] The molecular docking process was carried out by using GlideSP.sup.[1] in Maestro (Schrödinger version 2017-2) and the default options. The cocrystal structure with the PDB ID code of 6 WTT was selected as the docking template. For the preparation of protein, hydrogen atoms were added using the protein preparation wizard module of Maestro .sup.[2] and the OPLS3 force field was used. For the preparation of ligands, 3D structures were generated, and the energy minimization was performed by LigPrep.sup.[3]. A 30 Å docking grid was generated using the ligand centroids from the 7BV2 crystal structure. The ligands were then removed and embodiment compounds were placed during molecular docking. The type of interaction between protein receptor and ligand was analyzed, and then the reasonable docking conformation was selected and saved according to the calculated docking score and globalStrain values. The simulation results of compounds 15-27 binding to 6WTT protein are shown in FIGS. 1 to 13.

[0267] [1] Glide, Schrödinger, LLC, New York, NY, 2017.

[0268] [2] Maestro, Schrödinger, LLC, New York, NY, 2017.

[0269] [3] LigPrep, Schrödinger, LLC, New York, NY, 2017.

[0270] Conclusion: The compounds of the present disclosure have a good combination with 6WTT protein.

Embodiment 1

##STR00530##

[0271] Synthetic route:

##STR00531##

Step 1: Synthesis of Compound BB-1-2

[0272] Compound BB-1-1 (11 g, 38.42 mmol) was added to ammonia methanol solution (7 M, 54.88 mL), and the tube was sealed, and then the reaction was gradually heated to 50° C. and stirred continuously for 16 hours. The reaction mixture was concentrated under reduced pressure, and dissolved with an appropriate amount of DCM then concentrated again. The residue was not purified to obtain compound BB-1-2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ = 7.23 - 7.07 (m, 1H), 6.57 - 6.36 (m, 1H), 6.12 - 5.96 (m, 1H), 5.95 - 5.82 (m, 1H), 4.44 - 4.28 (m, 1H), 3.46 - 3.23 (m, 2H), 2.60 - 2.49 (m, 1H), 2.46 - 2.29 (m, 1H), 2.15 - 2.00 (m, 1H), 1.96 - 1.78 (m, 2H), 1.55 - 1.37 (m, 9H).

Step 2: Synthesis of Hydrochloride of Compound BB-1

[0273] Compound BB-1-2 (2 g, 7.37 mmol) was added to ethyl acetate (10 mL), and 4 M ethyl acetate solution (20 mL) of hydrogen chloride was added thereto, and then the reaction was stirred continuously at 20° C. for 3 hours. The reaction mixture was filtered to obtain a white solid, which was quickly transferred to a flask (highly susceptible to moisture absorption), and concentrated under reduced pressure. Hydrochloride of compound BB-1 was obtained. .sup.1HNMR (400 MHz, CD.sub.3OD) δ = 4.06 - 3.99 (m, 1H), 3.69 - 3.60 (m, 2H), 3.44 - 3.37 (m, 2H), 2.83 - 2.70 (m, 1H), 2.48 - 2.37 (m, 1H), 2.11 - 2.02 (m, 2H), 1.94 - 1.81 (m, 1H), 1.65 - 1.55 (m, 2H).

Step 3: Synthesis of Compound 1-2

[0274] At 0° C., toluene (4 mL), (trimethylsilyl)diazomethane (2 M, 1.66 mL) were added to a solution of compound 1-1 (400.00 mg, 1.66 mmol) in methanol (2 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 10:1) to obtain compound 1-2. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 4.39 - 4.20 (m, 1H), 3.87 - 3.68 (m, 4H), 2.74 -2.62 (m, 1H), 1.98 - 1.87 (m, 1H), 1.83 - 1.61 (m, 3H), 1.56 - 1.49 (m, 1H), 1.48 - 1.36 (m, 9H), 1.30 - 1.21 (m, 1H).

Step 4: Synthesis of Hydrochloride of Compound 1-3

[0275] Ethyl acetate hydrochloride (4 M, 5 mL) was added to a reaction flask with compound 1-2 (0.28 g, 1.10 mmol) and the reaction mixture was reacted at 20° C. for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product. Hydrochloride of compound 1-3 was obtained. .sup.1HNMR (400 MHz, CD.sub.3OD) δ = 4.21 - 4.07 (m, 2H), 3.94 - 3.79 (m, 3H), 3.03 - 2.91 (m, 1H), 2.04 - 2.03 (m, 1H), 2.05 - 2.00 (m, 1H), 1.93 - 1.80 (m, 3H), 1.73 (s, 2H).

Step 5: Synthesis of Compound 1-4

[0276] At 0° C., (S)-2-((tert-butoxycarbonyl)amino)-2-(4-fluorophenyl)acetic acid (0.5 g, 1.86 mmol), N,N-diisopropylethylamine (719.95 mg, 5.57 mmol) and 2-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (1.06 g, 2.79 mmol) were added to a solution of hydrochloride of compound 1-3 (427.06 mg, 2.23 mmol) in N,N-dimethylformamide (5 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was poured into 5% citric acid solution to separate the phases, then the aqueous phase was extracted with ethyl acetate (50 mL×2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=3:1) to obtain compound 1-4. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 7.60 - 7.33 (m, 2H), 7.16 - 6.93 (m, 2H), 5.77 (br d, J= 7.9 Hz, 1H), 5.50 - 5.36 (m, 1H), 3.87 - 3.68 (m, 3H), 2.81 - 2.59 (m, 1H), 1.88 - 1.68 (m, 3H), 1.60 (s, 1H), 1.48 - 1.39 (m, 9H), 1.34 - 1.19 (m, 4H).

Step 6: Synthesis of Compound 1-5

[0277] Lithium hydroxide monohydrate (148.66 mg, 3.54 mmol) was added to a solution of compound 1-4 (0.72 g, 1.77 mmol) in tetrahydrofuran (10 mL) and water (5 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was added with 50 mL of 5% citric acid aqueous solution, and 50 mL of ethyl acetate was added to separate the phases, then the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. Compound 1-5 was obtained. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 7.52 - 7.34 (m, 2H), 7.13 - 7.01 (m, 2H), 5.99 (br d, J= 7.3 Hz, 1H), 5.55 - 5.36 (m, 1H), 4.45 - 4.18 (m, 1H), 4.08 - 3.98 (m, 1H), 3.06 - 2.87 (m, 1H), 2.01 - 1.72 (m, 3H), 1.69 - 1.52 (m, 1H), 1.47 - 1.37 (m, 10H), 1.27 (br t, J= 7.1 Hz, 1H).

Step 7: Synthesis of Compound 1-6

[0278] At 0° C., hydrochloride of compound BB-1 (412.75 mg, 1.99 mmol), N,N-diisopropylethylamine (642.23 mg, 4.97 mmol, 865.54 .Math.L), 1-hydroxybenzotriazole (268.58 mg, 1.99 mmol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-acetaldehyde (381.04 mg, 1.99 mmol) were added to a solution of compound 1-5 (0.65 g, 1.66 mmol) in butanone (10 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was poured into 20 mL of water, and a mixed solution of dichloromethane and methanol (volume ratio of 5:1) (50 mL×2) was added for extraction, and the organic phase was washed with 5% citric acid solution (50 mL× 1) and washed with water (50 mL× 1), then the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (dichloromethane/methanol = 15:1) to obtain compound 1-6. .sup.1HNMR (400 MHz, DMSO-d.sub.6) δ = 8.52 - 7.98 (m, 1H), 7.71 - 7.58 (m, 1H), 7.54 - 7.39 (m, 2H), 7.29 - 7.08 (m, 3H), 7.03 (br s, 1H), 5.46 (br d, J= 8.5 Hz, 1H), 4.50 - 4.18 (m, 1H), 4.13 (br s, 1H), 3.51 (br s, 1H), 3.15 - 3.05 (m, 1H), 2.61 - 2.46 (m, 5H), 2.37 - 1.84 (m, 3H), 1.82 - 1.49 (m, 5H), 1.45 - 1.28 (m, 9H), 1.25 - 1.16 (m, 1H).

Step 8: Synthesis of Trifluoroacetate of Compound 1-7

[0279] At 0° C., trifluoroacetic acid (1 mL) was added to a solution of compound 1-6 (0.3 g, 515.41 .Math.mol) in dichloromethane (3 mL), and the reaction mixture was reacted at 20° C. for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product. Trifluoroacetate of compound 1-7 was obtained.

Step 9: Synthesis of Compound 1

[0280] At 0° C., pyridine (492.38 mg, 6.22 mmol, 502.43 .Math.L) and trifluoroacetic anhydride (272.37 mg, 1.30 mmol, 180.38 .Math.L) were added to a solution of trifluoroacetate of compound 1-7 (0.25 g, 518.73 .Math.mol) in tetrahydrofuran (2 mL), and the reaction mixture was reacted at 20° C. for 2 hours. The reaction mixture was quenched with 20 mL of water, and ethyl acetate (20 mL× 2) was added for extraction, then the organic phase was washed with 5% citric acid (20 mL×1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative HPLC (column type: Phenomenex C18 75*30 mm*3 .Math.m; mobile phase: [H.sub.2O (NH.sub.4HCO.sub.3)-ACN]; ACN%: 20%-60%, 8 min) to obtain compound 1. .sup.1HNMR (400 MHz, DMSO-d.sub.6) δ = 9.42 - 9.07 (m, 1H), 8.33 - 7.97 (m, 1H), 6.97 - 6.83 (m, 1H), 6.78 - 6.61 (m, 2H), 6.46 - 6.34 (m, 2H), 4.95 - 4.82 (m, 1H), 4.28 - 4.02 (m, 1H), 3.28 - 3.15 (m, 1H), 3.06 - 2.98 (m, 1H), 2.40 - 2.16 (m, 2H), 1.58 - 1.50 (m, 1H), 1.41 - 1.16 (m, 2H), 1.04 - 0.81 (m, 6H), 0.64 - 0.47 (m, 1H), 0.41 (br d, J= 9.3 Hz, 1H).

Embodiment 2

[0281] ##STR00532##

[0282] Synthetic route:

##STR00533##

Step 1: Synthesis of Compound 2-1

[0283] At 0° C., Boc-L-cyclohexylglycine (0.5 g, 2.61 mmol), N,N-diisopropylethylamine (1.01 g, 7.83 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N,N tetramethyluronium hexafluorophosphate (1.49 g, 3.92 mmol) were added to a solution of hydrochloride of compound 1-3 (671.62 mg, 2.61 mmol) in N,N-dimethylformamide (5 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was poured into 5% citric acid solution to separate the phases, then the aqueous phase was extracted with ethyl acetate (50 mL×2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 3:1) to obtain compound 2-1. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 5.16 (br d, J= 9.4 Hz, 1H), 4.48 - 4.34 (m, 1H), 4.32 - 4.23 (m, 1H), 4.08 - 4.00 (m, 1H), 3.71 (s, 3H), 2.77 - 2.69 (m, 1H), 2.05 - 1.99 (m, 1H), 1.91 - 1.62 (m, 11H), 1.45 - 1.41 (m, 9H), 1.23 - 1.03 (m, 4H).

Step 2: Synthesis of Compound 2-2

[0284] Lithium hydroxide monohydrate (212.72 mg, 5.07 mmol) was added to a solution of compound 2-1 (1 g, 2.53 mmol) in tetrahydrofuran (10 mL) and water (5 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was added with 50 mL of 5% citric acid aqueous solution, and 50 mL of ethyl acetate was added to separate the phases, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. Compound 2-2 was obtained. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 5.17 (br d, J= 9.7 Hz, 1H), 4.39 (br s, 1H), 4.35 - 4.24 (m, 1H), 4.16 - 4.11 (m, 1H), 3.04 - 2.93 (m, 1H), 1.96 (br d, J= 10.3 Hz, 1H), 1.87 - 1.53 (m, 11H), 1.44 (s, 9H), 1.19 - 0.95 (m, 4H).

Step 3: Synthesis of Compound 2-3

[0285] At 0° C., hydrochloride of compound BB-1 (438.81 mg, 2.11 mmol), N,N-diisopropylethylamine (682.77 mg, 5.28 mmol, 920.18 .Math.L), 1-hydroxybenzotriazole (285.53 mg, 2.11 mmol), hydrochloride of 1-(3-dimethylaminopropyl)-3-acetaldehyde (405.09 mg, 2.11 mmol) were added to a solution of compound 2-2 (0.65 g, 1.66 mmol) in butanone (10 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was poured into 20 mL of water, and extracted with a mixed solution of dichloromethane and methanol (dichloromethane: methanol = 10:1, 50 mL×2), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was separated by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound 2-3. .sup.1HNMR (400 MHz, DMSO-d.sub.6) δ = 8.08 (br d, J= 8.4 Hz, 1H), 7.75 - 7.55 (m, 1H), 7.32 - 7.18 (m, 1H), 7.08 - 6.94 (m, 1H), 6.87 - 6.64 (m, 1H), 4.42 - 4.31 (m, 1H), 4.20 (ddd, J= 3.8, 8.2, 11.6 Hz, 1H), 4.11 - 4.00 (m, 3H), 3.14 - 2.96 (m, 2H), 2.55 (br s, 1H), 2.42 - 2.32 (m, 1H), 2.21 - 2.09 (m, 1H), 2.04 - 1.95 (m, 1H), 1.93 - 1.81 (m, 1H), 1.78 - 1.49 (m, 11H), 1.40 - 1.24 (m, 11H), 1.14 - 0.86 (m, 5H).

Step 4: Synthesis of Trifluoroacetate of Compound 2-4

[0286] At 0° C., trifluoroacetic acid (5 mL) was added to a solution of compound 2-3 (0.86 g, 1.61 mmol) in dichloromethane (15 mL), and the reaction mixture was reacted at 20° C. for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product. Trifluoroacetate of compound 2-4 was obtained.

Step 5: Synthesis of Compound 2

[0287] At 0° C., pyridine (1.41 g, 17.87 mmol) and trifluoroacetic anhydride (782.03 mg, 3.72 mmol) were added to a solution of trifluoroacetate of compound 2-4 (0.7 g, 1.49 mmol) in tetrahydrofuran (10 mL), and the reaction mixture was reacted at 20° C. for 2 hours. The reaction mixture was added with 20 mL of water for quenching, and ethyl acetate (20 mL× 2) was added for extraction, then the organic phase was washed with 5% citric acid (20 mL×1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative HPLC (column type: Phenomenex C18 75*30 mm*3 .Math.m; mobile phase: [H.sub.2O (NH.sub.4HCO.sub.3)-ACN]; ACN%: 25%-65%,8 min) to obtain compound 2. .sup.1HNMR (400 MHz, DMSO-.sub.d6) δ = 9.67 (br d, J= 7.2 Hz, 1H), 8.90 - 8.74 (m, 1H), 7.76 - 7.60 (m, 1H), 5.05 - 4.88 (m, 1H), 4.56 - 4.46 (m, 1H), 4.45 - 4.29 (m, 1H), 3.75 (s, 1H), 3.18 - 3.02 (m, 2H), 2.46 - 2.38 (m, 1H), 2.16 - 2.04 (m, 3H), 1.87 - 1.59 (m, 11H), 1.39 - 1.26 (m, 2H), 1.16 - 0.95 (m, 5H).

Embodiment 3

[0288] ##STR00534##

[0289] Synthetic route:

##STR00535##

Step 1: Synthesis of Compound 3-2

[0290] Compound 3-1 (1 g, 3.23 mmol) was dissolved in N,N-dimethylformamide (10 mL), and O-(7-azabenzotriazol-1-yl)-N,N,N,N tetramethyluronium hexafluorophosphate (1.84 g, 4.85 mmol) was added thereto. The reaction was stirred at 20° C. for 0.5 hours, then diisopropylethylamine (2.09 g, 16.16 mmol) and hydrochloride of compound 1-3 (601.92 mg, 3.88 mmol) were added and the reaction mixture was reacted at 20° C. for 16 hours. The reaction system was added with ethyl acetate (50 mL), and added with 3% citric acid solution (25 mL) and saturated brine (25 mL) in turn for extraction to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to obtain compound 3-2. .sup.1HNMR (400 MHz, CD.sub.3OD) δ = 6.27 (br d, J=9.6 Hz, 1H), 4.62 (br s, 1H), 4.19 (d, J=9.6 Hz, 1H), 3.99 (s, 1H), 3.74 - 3.68 (m, 3H), 2.71 (br s, 1H), 1.92 - 1.49 (m, 21H), 1.44 (s, 9H).

Step 2: Synthesis of Compound 3-3

[0291] Compound 3-2 (1.2 g, 2.69 mmol) was dissolved in tetrahydrofuran (9 mL) and water (4 mL), and lithium hydroxide monohydrate (225.52 mg, 5.37 mmol) was added thereto, and the reaction mixture was stirred at 20° C. for 16 hours. The reaction system was added with ethyl acetate (60 mL) and 3% citric acid (30 mL) for extraction to separate the organic phase, and the organic phase was washed with saturated brine (30 mL×2) to neutral, dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 3-3 was obtained. .sup.1HNMR (400 MHz, CD.sub.3OD) δ = 4.60 (br s, 1H), 4.22 - 4.15 (m, 1H), 3.95 (s, 1H), 2.74 (br s, 1H), 2.05 - 1.94 (m, 6H), 1.87 - 1.61 (m, 15H), 1.46 - 1.40 (m, 9H).

Step 3: Synthesis of Compound 3-4

[0292] Compound 3-3 (1.1 g, 2.54 mmol) was dissolved in 2-butanone (12 mL), then 1-hydroxybenzotriazole (343.62 mg, 2.54 mmol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (585.01 mg, 3.05 mmol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours. Then diisopropylethylamine (1.64 g, 12.72 mmol) and hydrochloride of compound BB-1 (580.89 mg, 2.80 mmol) were added thereto, and the reaction mixture was stirred continuously at 20° C. for 16 hours. The reaction system was added with dichloromethane (60 mL) and 3% citric acid (30 mL) for extraction to separate the organic phase, and the organic phase was washed with saturated brine (30 mL×2) to neutral to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation. The residue was purified by silica gel column chromatography (dichloromethane: methanol = 1:0 to 10:1) to obtain compound 3-4. .sup.1H NMR (400 MHz, CD.sub.3OD) δ = 4.58 (br s, 1H), 4.51 - 4.41 (m, 1H), 4.20 (s, 1H), 3.95 (s, 1H), 3.36 - 3.30 (m, 2H), 2.72 (br s, 1H), 2.69 - 2.59 (m, 1H), 2.43 - 2.30 (m, 1H), 2.22 (br d, J=9.8 Hz, 1H), 2.15 - 2.07 (m, 1H), 1.97 (br s, 4H), 1.77 - 1.61 (m, 18H), 1.44 (s, 9H).

Step 4: Synthesis of Trifluoroacetate of Compound 3-5

[0293] Compound 3-4 (1.1 g, 1.88 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (5 mL) was added thereto, and the reaction mixture was stirred at 20° C. for 1 hour, and the reaction system was directly evaporated to dryness by an oil pump, added with a small amount of dichloromethane and evaporated by rotary evaporation, and the above steps were repeated until the shape of the product was a white foam. Trifluoroacetate of compound 3-5 was obtained. .sup.1HNMR (400 MHz, CD.sub.3OD) δ = 4.55 (s, 1H), 4.49 (dd, J=3.6, 11.9 Hz, 1H), 4.02 (s, 1H), 3.85 (s, 1H), 3.30 - 3.19 (m, 2H), 2.77 (br s, 1H), 2.72 -2.63 (m, 1H), 2.45 - 2.31 (m, 1H), 2.23 (br d, J=10.0 Hz, 1H), 2.17 - 2.10 (m, 1H), 2.04 (br s, 4H), 1.94 -1.67 (m, 21H). [M+1].sup.+ = 486.3.

Step 5: Synthesis of Compound 3

[0294] Trifluoroacetate of compound 3-5 (900 mg, 1.85 mmol) was dissolved in tetrahydrofuran (10 mL), then pyridine (1.47 g, 18.53 mmol, 1.50 mL) and trifluoroacetic anhydride (973.13 mg, 4.63 mmol, 644.46 .Math.L) were added thereto at 0° C., and the temperature was slowly raised to 20° C., then the reaction mixture was stirred for 16 hours. The reaction system was added with dichloromethane (50 mL) and 3% citric acid (25 mL) for extraction to separate the organic phase, and the organic phase was extracted with saturated brine (25 mL) to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the crude product was separated by preparative HPLC to obtain compound 3. .sup.1HNMR (400 MHz, CD.sub.3OD) δ = 5.06 (dd, J=4.7, 11.3 Hz, 1H), 4.66 (s, 1H), 4.61 (s, 1H), 3.87 (s, 1H), 3.32 - 3.18 (m, 2H), 2.73 - 2.63 (m, 2H), 2.39 - 2.27 (m, 3H), 2.01 (br s, 4H), 1.84 (br s, 2H), 1.81 (br s, 2H), 1.77 -1.65 (m, 11H), 1.63 - 1.41 (m, 3H).

Embodiment 4

[0295] ##STR00536##

[0296] Synthetic route:

##STR00537##

Step 1: Synthesis of Compound 4-2

[0297] Compound 4-1 (2.74 g, 17.76 mmol) was dissolved in tetrahydrofuran (27.4 mL), and the reaction system was replaced with nitrogen for three times, cooled to 0° C., then a solution of borane in tetrahydrofuran (35.52 mL, 1 M) was slowly added dropwise thereto. The reaction was stirred at 20° C. for 16 hours, added with sodium hydroxide solution (80 mL, 1 M) at 0° C., extracted twice with methyl tert-butyl ether (100 mL), and the organic phases were combined and washed with 10% citric acid (80 mL×2) and saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered and concentrated. Compound 4-2 was obtained. .sup.1H NMR (400 MHz, CDCl.sub.3) δ ppm 3.19 (s, 2 H) 1.50 - 1.59 (m, 7 H) 1.29 - 1.38 (m, 6 H).

Step 2: Synthesis of Compound 4-3

[0298] Compound 4-2 (2.26 g, 16.09 mmol) was dissolved in dichloromethane (67.8 mL), and Dess-Martin periodinane (10.24 g, 24.14 mmol) was added to the reaction system. The reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with sodium thiosulfate (50 mL) and saturated sodium bicarbonate solution (70 mL), and extracted twice with dichloromethane (100 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 4-3 was obtained. .sup.1HNMR (400 MHz, CDCl.sub.3) δ ppm 9.41 (s, 1 H) 1.60 (s, 13 H).

Step 3: Synthesis of Compound 4-4

[0299] Compound 4-3 (1.97 g, 14.25 mmol) was dissolved in methanol (137.9 mL), and compound R-phenylglycinol (2.35 g, 17.10 mmol) was added to the reaction system, and the reaction mixture was stirred at 20° C. for 2 hours. The reaction mixture was cooled to 0° C., added with trimethylsilyl cyanide (9.90 g, 99.78 mmol), and stirred at 50° C. for 16 hours. The reaction mixture was directly evaporated to dryness by rotary evaporation. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound 4-4. .sup.1HNMR (400 MHz, CDCl.sub.3) δ ppm 7.28 - 7.39 (m, 5 H) 4.06 (dd, J=9.10, 4.06 Hz, 1 H) 3.80 (dd, J=10.96, 3.95 Hz, 1 H) 3.58 (t, J=9.98 Hz, 1 H) 2.89 (s, 1H) 1.56 - 1.65 (m, 13 H).

Step 4: Synthesis of Hydrochloride of Compound 4-5

[0300] Compound 4-4 (1.42 g, 4.99 mmol) was dissolved in hydrochloric acid (28.4 mL) and glacial acetic acid (7.1 mL), and the reaction was stirred at 80° C. for 16 hours. The reaction mixture was cooled to 0° C. to precipitate the solid, and filtered. Hydrochloride of compound 4-5 was obtained. [M+1].sup.+ = 303.2.

Step 5: Synthesis of Hydrochloride of Compound 4-6

[0301] Hydrochloride of compound 4-5 (2.24 g, 7.38 mmol) was dissolved in methanol (112 mL) and glacial acetic acid (22.4 mL), then 20% wet palladium hydroxide (0.448 g, 638.02 .Math.mol) was added thereto. 50 psi hydrogen was introduced to the reaction system, and the reaction mixture was heated to 50° C. and stirred for 18 hours. The reaction mixture was filtrated and directly evaporated to dryness by rotary evaporation, and slurried with methyl tert-butyl ether (40 mL), and filtered. Hydrochloride of compound 4-6 was obtained. .sup.1HNMR (400 MHz, CD.sub.3OD) δ ppm 3.53 (s, 1 H) 1.47 - 1.71 (m, 13 H) 3.53 (s, 1 H).

Step 6: Synthesis of Compound 4-7

[0302] Hydrochloride of compound 4-6 (50 mg, 272.86 .Math.mol) was dissolved in 1,4- dioxane (0.375 mL) and water (1 mL), then anhydrous sodium carbonate (115.68 mg, 1.09 mmol) and di-tert-butyl dicarbonate (119.10 mg, 545.71 .Math.mol) were added thereto, and the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (5 mL) and 5% citric acid (10 mL), and extracted twice with ethyl acetate (20 mL), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation. Compound 4-7 was obtained. .sup.1HNMR (400 MHz, CDCl.sub.3) δ ppm 4.98 (br d, J=8.63 Hz, 1 H) 3.97 - 4.05 (m, 1 H) 1.43 - 1.62 (m, 22 H).

Step 7: Synthesis of Compound 4-8

[0303] Compound 4-7 (250 mg, 882.26 .Math.mol) was dissolved in N,N-dimethylformamide (2.5 mL), then 1-hydroxybenzotriazole (357.64 mg, 2.65 mmol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (338.26 mg, 1.76 mmol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours, then N,N-diisopropylethylamine (342.08 mg, 2.65 mmol) and hydrochloride of compound 1-3 (136.92 mg, 882.26 .Math.mol) were added thereto, and the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (10 mL), extracted twice with ethyl acetate (20 mL), then the organic phases were combined, washed twice with 5% citric acid (15 mL), and washed four times with brine (10 mL), then dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (petroleum ether: ethyl acetate=8:1) to obtain compound 4-8. .sup.1HNMR (400 MHz, CDCl.sub.3) δ ppm 5.20 (br d, J=9.63 Hz, 1 H) 4.48 - 4.54 (m, 1 H) 4.22 - 4.30 (m, 1 H) 4.02 - 4.11 (m, 1 H) 3.76 (s, 3 H) 2.76 (br s, 1 H) 2.05 (br d, J=10.38 Hz, 1 H) 1.68 - 1.84 (m, 4 H) 1.60 (s, 13 H) 1.46 (s, 9 H).

Step 8: Synthesis of Compound 4-9

[0304] Compound 4-8 (300 mg, 713.37 .Math.mol) was dissolved in tetrahydrofuran (3 mL) and water (1 mL), then lithium hydroxide monohydrate (59.87 mg, 1.43 mmol) was added thereto and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with water (10 mL) and 5% citric acid (15 mL), extracted twice with ethyl acetate (15 mL). The organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. Compound 4-9 was obtained without purification. .sup.1HNMR (400 MHz, CDCl.sub.3) δ ppm 5.07 - 5.17 (m, 1 H) 4.42 - 4.47 (m, 1 H) 4.26 (br d, J=9.88 Hz, 1H) 4.17 (s, 1H) 3.07 (br s, 1 H) 1.70-1.98 (m, 5 H) 1.46 - 1.67 (m, 13 H).

Step 9: Synthesis of Compound 4-10

[0305] Compound 4-9 (266 mg, 654.36 .Math.mol) was dissolved in 2-butanone, then 1-hydroxybenzotriazole (88.42 mg, 654.34 .Math.mol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (150.53 mg, 785.21 .Math.mol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours, and N,N-diisopropylethylamine (338.28 mg, 2.62 mmol) and hydrochloride compound BB-1 (134.42 mg, 785.21 .Math.mol) were added thereto, and the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (10 mL), extracted twice with dichloromethane (20 mL), and the organic phases were combined and washed twice with 5% citric acid (15 mL), washed twice with brine (10 mL), and then dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (dichloromethane: methanol =20:1) to obtain compound 4-10. [M+1].sup.+ = 560.4.

Step 10: Synthesis of Trifluoroacetate of Compound 4-11

[0306] Compound 4-10 (166 mg, 296.59 .Math.mol) was dissolved in dichloromethane (1.8 mL) and trifluoroacetic acid (0.6 mL), and the reaction was stirred at 20° C. for 2 hours. The reaction was directly evaporated to dryness by an oil pump, added with a small amount of dichloromethane and evaporated by rotary evaporation, and the above steps were repeated until the shape of the product was a light yellow foam. Trifluoroacetate of compound 4-11 was obtained. [M+1].sup.+ = 460.4.

Step 11: Synthesis of Compound 4

[0307] Trifluoroacetate of compound 4-11 (136 mg, 295.92 .Math.mol) was dissolved in tetrahydrofuran (1.4 mL), cooled to 0° C., then pyridine (79.10 mg, 2.07 mmol) and trifluoroacetic anhydride (210.03 mg, 1.18 mmol) were added thereto. The reaction was raised to room temperature of 20° C. and stirred for 16 hours. The reaction mixture was extracted twice with water (10 mL) and extracted twice with dichloromethane (10 mL), and the organic phases were combined and washed twice with 3% citric acid (10 mL), and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by preparative HPLC (column type: C18-2 100*30 mm*5 .Math.m; mobile phase: [H.sub.2O (NH.sub.4HCO.sub.3)-ACN]; ACN%: 30%-50%, 20 min) to obtain compound 4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ = 9.39 - 8.06 (m, 1H), 7.14 - 6.89 (m, 1H), 6.10 - 5.81 (m, 1H), 4.99 - 4.69 (m, 1H), 4.63 - 4.39 (m, 1H), 4.03 - 3.86 (m, 1H), 3.48 - 3.26 (m, 2H), 2.98 - 2.79 (m, 1H), 2.61 - 1.17 (m, 25H).

Embodiment 5

##STR00538##

[0308] Synthetic route:

Step 1: Synthesis of Compound 5-2

[0309] Compound 5-1 (5 g, 54.32 mmol) was dissolved in methanol (50 mL), and refluxed at 70° C. for 48 hours. The reaction system was concentrated under reduced pressure to obtain a crude product of the target product. The crude product with high purity was directly used in the next reaction to obtain compound 5-2. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 4.81 (s, 1H), 3.77 (s, 3H), 3.43 (s, 3H).

Step 2: Synthesis of Compound 5-3

[0310] Compound 5-2 was dissolved in toluene (3 mL), cooled to 0° C., and compound (R)-(+)-phenethylamine (1.5 g, 12.38 mmol, 1.60 mL) was slowly added dropwise thereto, and the reaction mixture was stirred at 20° C. for 1 hour. The reaction system was added with ethyl acetate (60 mL) and saturated brine (30 mL) for extraction to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate and evaporated to dryness by rotary evaporation to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to obtain target compound 5-3. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 7.95 - 7.56 (m, 1H), 7.31 - 7.17 (m, 5H), 4.71 - 4.40 (m, 1H), 3.95 - 3.71 (m, 3H), 1.67 - 1.51 (m, 3H).

Step 3: Synthesis of Compound 5-4

[0311] Compound 5-3 (0.5 g, 2.61 mmol) was dissolved in 2,2,2-trifluoroethanol (5 mL), and trifluoroacetic acid (313.04 mg, 2.75 mmol, 203.28 .Math.L) was added thereto, cooled to -10° C., stirred for 1 hour, and the temperature was controlled to -10° C., and cyclopentadiene (207.40 mg, 3.14 mmol) was slowly added dropwise thereto, and the reaction mixture was stirred continuously for 0.5 hours. The reaction system was concentrated under reduced pressure, then methyl tert-butyl ether (60 mL) and saturated sodium bicarbonate solution (30 mL×2) were added thereto, stirred for 10 minutes, extracted to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to obtain compound 5-4. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 7.34 - 7.18 (m, 5H), 6.59 -6.41 (m, 1H), 6.31 (dd, J= 1.6, 5.6 Hz, 1H), 4.35 (brd, J= 1.3 Hz, 1H), 3.39 (s, 3H), 3.18 - 3.03 (m, 1H), 2.95 (br s, 1H), 2.33 - 2.22 (m, 1H), 2.14 (brd, J= 8.4 Hz, 1H), 1.54 - 1.41 (m, 4H). [M+1].sup.+ =258.2.

Step 4: Synthesis of Compound 5-5

[0312] Compound 5-4 (100.00 mg, 388.61 .Math.mol) was dissolved in tetrahydrofuran (1.25 mL), cooled to -70° C., and borane tetrahydrofuran complex (1 M, 427.47 .Math.L) was slowly added dropwise thereto and the reaction mixture was slowly raised to 20° C. and stirred for 1 hour. The reaction mixture was cooled to 0° C., and 10% sodium hydroxide aqueous solution (0.55 mL) and 30% hydrogen peroxide (220.28 mg, 1.94 mmol, 186.68 .Math.L) solution were added thereto, then the temperature was slowly raised to 20° C., and the reaction mixture was stirred for 1 hour. The reaction system was added with saturated sodium thiosulfate solution (10 mL) and stirred for 10 minutes to quench, and saturated brine (20 mL) and ethyl acetate (60 mL×2) were added for extraction to separate the organic phase. A small amount of sample solution was taken, and the pH was adjusted with 3% citric acid to less than 8, and after starch potassium iodide test paper showed negative results, the mixture was dried over anhydrous sodium sulfate and concentrated at 30° C. under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to obtain compound 5-5. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 7.30 - 7.13 (m, 5H), 3.93 (br d, J= 6.5 Hz, 1H), 3.78 (br s, 1H), 3.70 - 3.54 (m, 1H), 3.39 - 3.32 (m, 1H), 3.31 - 3.24 (m, 3H), 2.49 - 2.40 (m, 1H), 2.26 (s, 1H), 2.09 - 2.00 (m, 1H), 1.72 (br d, J= 10.1 Hz, 1H), 1.46 (br d, J= 6.5 Hz, 1H), 1.41 -1.33 (m, 3H). [M+1].sup.+=276.1.

Step 5: Synthesis of Compound 5-6

[0313] Compound 5-5 (800 mg, 2.91 mmol) was dissolved in acetonitrile (10 mL), and 2-iodoxybenzoic acid (976.31 mg, 3.49 mmol) was added thereto and the reaction mixture was reacted at 75° C. for 1 hour. The reaction system was dried over anhydrous sodium sulfate, filtered, and the filter cake was washed with acetonitrile (10 mL), and the filtrates were combined and concentrated to obtain compound 5-6. [M+1].sup.+ =274.0.

Step 6: Synthesis of Compound 5-7

[0314] Compound 5-6 was dissolved in tetrahydrofuran (20 mL) and cooled to 0° C., and under nitrogen atmosphere, 0.5 M toluene solution of bis(cyclopentadienyl)-.Math.-chloro(dimethylaluminum)-.Math.-methylenetitanium (0.5 M, 4.02 mL) was slowly added dropwise thereto, and the temperature was slowly raised to 20° C., and the reaction mixture was stirred for 2.5 hours. The temperature was lowered to 0° C., and saturated sodium bicarbonate solution (1 mL) was added to the reaction system for quenching, then methyl tert-butyl ether (10 mL) was added thereto, and anhydrous sodium sulfate was added to dry, and the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to obtain compound 5-7. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 7.28 - 7.09 (m, 5H), 4.92 (br s, 1H), 4.71 (s, 1H), 3.81 (s, 1H), 3.48 (q, J= 6.5 Hz, 1H), 3.28 -3.15 (m, 3H), 2.76 - 2.69 (m, 2H), 2.65 (br d, J= 16.8 Hz, 1H), 2.21 (br d, J= 9.8 Hz, 1H), 2.08 - 1.95 (m, 1H), 1.43 (d, J= 9.8 Hz, 1H), 1.31 (d, J= 6.5 Hz, 3H). [M+1].sup.+=272.0.

Step 7: Synthesis of Hydrochloride of Compound 5-8

[0315] Compound 5-7 (310 mg, 1.14 mmol) was dissolved in ethanol (5 mL), and 12 M of hydrochloric acid (285.61 .Math.L) and wet palladium on carbon (1 g, palladium content of 10%) were added thereto, and the reaction mixture was reacted under a hydrogen balloon atmosphere at 15 psi and 20° C. for 16 hours. The reaction system was filtered through diatomite and the filtrate was evaporated to dryness by rotary evaporation to obtain hydrochloride of compound 5-8. .sup.1HNMR (400 MHz, CD.sub.3OD) δ = 4.17 - 4.00 (m, 1H), 3.88 - 3.81 (m, 3H), 3.74 - 3.41 (m, 1H), 2.55 - 2.20 (m, 1H), 2.17 - 1.99 (m, 1H), 1.96 - 1.71 (m, 3H), 1.36 - 1.23 (m, 2H), 1.18 - 1.01 (m, 3H). [M+1].sup.+=170.0.

Step 8: Synthesis of Compound 5-9

[0316] Compound 1-b (311.63 mg, 1.35 mmol) was dissolved in N,N-dimethylformamide (3 mL), then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (640.38 mg, 1.68 mmol) and diisopropylethylamine (725.55 mg, 5.61 mmol, 977.83 .Math.L) were added thereto, and the reaction mixture was stirred for 30 minutes, then hydrochloride of compound 5-8 (190 mg, 1.12 mmol) was added thereto, and the reaction mixture was stirred continuously at 20° C. for 2.5 hours. The reaction system was added with ethyl acetate (60 mL) and 3% citric acid solution (30 mL) for extraction to separate the organic phase, and the organic phase was extracted with saturated brine (30 mL) to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EA=1:0 to 10:1) to obtain compound 5-9. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 6.62 - 6.36 (m, 1H), 5.26 - 5.12 (m, 1H), 4.68 - 4.63 (m, 1H), 4.34 - 4.26 (m, 1H), 3.74 - 3.63 (m, 3H), 2.30 - 2.18 (m, 1H), 2.11 - 2.02 (m, 1H), 1.62 -1.52 (m, 4H), 1.50 (br s, 3H), 1.13 -1.07 (m, 9H), 1.05 (br s, 9H). [M+1].sup.+=383.3.

Step 9: Synthesis of Compound 5-10

[0317] Compound 5-9 (0.2 g, 522.89 .Math.mol) was dissolved in tetrahydrofuran (1.5 mL) and water (0.5 mL), and lithium hydroxide monohydrate (43.88 mg, 1.05 mmol) was added thereto, and the reaction mixture was reacted at 20° C. for 16 hours. The reaction system was added with dichloromethane (60 mL) and 3% citric acid (30 mL) for extraction to separate the organic phase, and the organic phase was added with saturated brine (30 mL) for extraction to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 5-10. [M+1].sup.+ =369.3.

Step 10: Synthesis of Compound 5-11

[0318] Compound 5-10 (190 mg, 515.65 .Math.mol) was dissolved in N,N-dimethylformamide (2 mL), then 1-hydroxybenzotriazole (69.67 mg, 515.65 .Math.mol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (118.62 mg, 618.78 .Math.mol) were added thereto, and the reaction mixture was stirred for 30 minutes, then diisopropylethylamine (333.21 mg, 2.58 mmol, 449.07 .Math.L) and hydrochloride of compound BB-1 (128.49 mg, 618.78 .Math.mol) were added thereto, and the reaction mixture was reacted at 20° C. for 2 hours. The reaction system was added with ethyl acetate (50 mL) and 3% citric acid (25 mL) for extraction to separate the organic phase, and the organic phase was extracted with saturated brine (25 mL) to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol = 1:0 to 30:1) to obtain compound 5-11. [M+1].sup.+=522.4.

Step 11: Synthesis of Trifluoroacetate of Compound 5-12

[0319] Compound 5-11 was dissolved in dichloromethane (1 mL), then trifluoroacetic acid (0.3 mL) was added thereto, and the reaction mixture was reacted at 20° C. for 1 hour. The reaction system was concentrated under reduced pressure to obtain trifluoroacetate of compound 5-12. [M+1].sup.+=422.3.

Step 12: Synthesis of Compound 5

[0320] Trifluoroacetate of compound 5-12 (80 mg) was dissolved in dichloromethane (1 mL), then pyridine (150.12 mg, 1.90 mmol, 153.18 .Math.L) and trifluoroacetic anhydride (99.65 mg, 474.46 .Math.mol, 65.99 .Math.L) were added thereto at 0° C., and the reaction mixture was slowly raised to 20° C. and reacted for 2 hours. The reaction system was added with 3% citric acid solution (30 mL) and dichloromethane (60 mL×2) for extraction to separate the organic phase, and the organic phase was washed with saturated brine (60 mL) until neutral, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol=10:1), and then purified by preparative high performance liquid chromatography (column type: Phenomenex Luna 80*30 mm*3 .Math.m; mobile phase: [water (hydrochloric acid)-acetonitrile]; acetonitrile%: 20%-40%, 8 min) to obtain compound 5. .sup.1HNMR (400 MHz, CDCl.sub.3) δ = 9.55 - 8.15 (m, 1H), 7.21 - 7.02 (m, 1H), 6.63 - 5.95 (m, 1H), 4.66 (br d, J= 6.0 Hz, 5H), 3.72 - 3.31 (m, 2H), 2.33 - 1.33 (m, 9H), 1.19 - 1.02 (m, 12H). [M+1].sup.+=500.3.

Embodiment 6

[0321] ##STR00539##

##STR00540##

[0322] Synthetic route:

Step 1: Synthesis of Compound 6-2

[0323] Compound 6-1 (30 g, 198.46 mmol) and glyoxylic acid (18.27 g, 198.46 mmol, 3.68 mL) were dissolved in anhydrous toluene (300 mL), and the reaction mixture was stirred at 40° C. for 3 hours. The reaction mixture was filtered and concentrated to obtain compound 6-2. [M+1].sup.+ = 226.2.

Step 2: Synthesis of Compound 6-3

[0324] Compound 6-2 was dissolved in anhydrous methanol (420 mL), and sulfuric acid (18.73 g, 190.94 mmol, 10.18 mL) was added thereto, and the reaction mixture was stirred at 20° C. for 48 hours. The reaction mixture was added with methyl tert-butyl ether (300 mL) and water (150 mL) for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product of the target product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound 6-3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 8.52 -8.46 (m, 1H), 7.40 - 7.30 (m, 6H), 5.08 (s, 2H), 3.68 - 3.64 (m, 3H), 3.26 (s, 3H).

Step 3: Synthesis of Compound 6-4

[0325] Compound 6-3 (26 g, 102.67 mmol) was dissolved in sulfuric acid (10.07 g, 102.67 mmol, 5.47 mL), and the mixture was added to anhydrous toluene (260 mL), heated to 70° C. Phosphorus trichloride (49.35 mg, 359.33 .Math.mol) was slowly added dropwise thereto, and the reaction mixture was stirred continuously at 75° C. for 16 hours. Then, the reaction mixture was concentrated under reduced pressure, and added with anhydrous toluene (200 mL) for dilution, and then evaporated by rotary evaporation under reduced pressure, which was repeated for three times. Finally, anhydrous toluene (200 mL) was added, and trimethyl phosphite (15.29 g, 123.20 mmol, 14.56 mL) was slowly added dropwise to the concentrated solution at 75° C., and then heated to 90° C. and reacted for 1.5 hours. The reaction mixture was poured into saturated sodium bicarbonate aqueous solution (360 mL) for quenching, added with ethyl acetate (360 mL) to separate the phases, and the organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound 6-4. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 7.41 - 7.30 (m, 5H), 5.09 -5.07 (m, 2H), 4.89 - 4.83 (m, 1H), 3.73 - 3.63 (m, 9H).

Step 4: Synthesis of Compound 6-6

[0326] At 20° C., compound 6-4 (20 g, 60.38 mmol) was dissolved in acetonitrile (50 mL), and 1,8-diazabicyclo[5.4.0]undec-7-ene (11.03 g, 72.45 mmol, 10.92 mL) was added thereto, and the reaction mixture was stirred for 0.5 hours, then a solution of compound 6-5 (6.65 g, 60.38 mmol) in acetonitrile (20 mL) was added thereto and the reaction mixture was stirred for 16 hours. Ethyl acetate (40 mL) and water (40 mL) were added to the reaction mixture to separate the phases, and the organic phases were combined and evaporated by rotary evaporation under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound 6-6. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 7.41 - 7.30 (m, 5H), 6.12 (br s, 1H), 5.13 (s, 2H), 3.77 - 3.71 (m, 3H), 3.11 (s, 2H), 2.87 - 2.81 (m, 2H), 2.10 (br s, 4H), 1.89 - 1.79 (m, 2H).

Step 5: Synthesis of Hydrochloride of Compound 6-7

[0327] Compound 6-6 (11 g, 34.88 mmol) was dissolved in anhydrous methanol (5 mL), and 10% wet palladium/carbon (2.20 g, 7.33 mol) was added thereto, then 15 psi hydrogen was introduced, and the reaction mixture was stirred at 30° C. for 18 hours. The reaction mixture was filtered through diatomite, and then concentrated under reduced pressure to obtain a crude product. The crude product was not further purified. Hydrochloride of compound 6-7 was obtained. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 3.60 - 3.57 (m, 3H), 3.32 (s, 2H), 3.17 - 3.12 (m, 1H), 1.98 - 1.73 (m, 11H).

Step 6: Synthesis of Compound 6-8

[0328] Compound 6-7 (6 g, 32.74 mmol) was added to a solution of water (30 mL) and anhydrous tetrahydrofuran (30 mL), then potassium carbonate (13.58 g, 98.23 mmol) and Boc anhydride (21.44 g, 98.23 mmol, 22.57 mL) were added thereto, and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with 5% citric acid (15 mL), and then added with water (40 mL), extracted with ethyl acetate (40 mL) to separate the phases. The aqueous phase was extracted with ethyl acetate (30 mL), then the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether: ethyl acetate=5:1) to obtain compound 6-8. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 3.84 -3.83 (m, 1H), 3.57 - 3.52 (m, 1H), 1.99 - 1.91 (m, 12H), 1.87 - 1.76 (m, 11H).

Step 7: Synthesis of Compound 6-9

[0329] Compound 6-8 (0.8 g, 2.82 mmol) was dissolved in a solution of anhydrous tetrahydrofuran (6 mL) and water (2 mL), and lithium hydroxide monohydrate (236.95 mg, 5.65 mmol) was added to the reaction mixture, then the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with ethyl acetate (5 mL), washed with water (5 mL), then the aqueous phase was added with 5% citric acid (15 mL) and then added with water (5 mL), and extracted with ethyl acetate (5 mLx2). The organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was not further purified to obtain compound 6-9. [M+1].sup.+ = 270.34.

Step 8: Synthesis of Compound 6-10

[0330] Compound 6-9 (3 g, 11.14 mmol) was added to N,N-dimethylformamide (30 mL), then 1-hydroxybenzotriazole (6.02 g, 44.55 mmol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (5.80 g, 44.90 mmol, 7.82 mL) were added thereto, and the reaction was stirred at 20° C. for 0.5 hours, then N,N-diisopropylethylamine (6.41 g, 33.42 mmol) and hydrochloride of compound 1-3 (2.13 g, 13.76 mmol) were added thereto, and the reaction mixture was stirred at 20° C. for 20 hours. The reaction mixture was added with 5% citric acid (15 mL), then added with water (40 mL), and extracted with ethyl acetate (40 mL). Then the aqueous phase was extracted with ethyl acetate (30 mL), and the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether: ethyl acetate=1:1) to obtain compound 6-10. [M+1].sup.+ = 406.25.

Step 9: Synthesis of Compound 6-11

[0331] Compound 6-10 (300.00 mg, 737.98 .Math.mol) was added to a mixture of anhydrous tetrahydrofuran (2.5 mL) and water (1 mL), and then lithium hydroxide monohydrate (61.94 mg, 1.48 mmol) was added to the reaction system, and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with ethyl acetate (10 mL), washed with water (10 mL), then the aqueous phase was added with 5% citric acid (2 mL), extracted with ethyl acetate (10 mLx2). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. Compound 6-11 was obtained from the crude product without further purification. [M+1].sup.+ = 392.23.

Step 10: Synthesis of Compound 6-12

[0332] Compound 6-11 (300.00 mg, 764.35 .Math.mol) was added to 2-butanone (6 mL), then 1-hydroxybenzotriazole (103.28 mg, 764.35 .Math.mol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (175.83 mg, 917.23 .Math.mol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours. N,N-Diisopropylethylamine (263.43 mg, 2.04 mmol, 355.03 .Math.L) and hydrochloride of compound BB-1 (158.72 mg, 764.35 .Math.mol) were added to the reaction system and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with 5% citric acid (5 mL), then added with water (10 mL), and extracted with ethyl acetate (10 mLx2). The aqueous phase was extracted with ethyl acetate (10 mL), and the organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane: anhydrous methanol = 10:1) to obtain compound 6-12. [M+1].sup.+ =545.32.

Step 11: Synthesis of Compound 6-13

[0333] Compound 6-12 (0.1 g, 183.26 .Math.mol) was dissolved in dichloromethane (1 mL), then trifluoroacetic acid (41.79 mg, 366.52 .Math.mol, 27.14 .Math.L) was added thereto, and the reaction mixture was stirred at 20° C. for 1 hour. The reaction mixture was then evaporated to dryness by rotary evaporation, added with dichloromethane and concentrated under reduced pressure, and the above steps were repeated for three times, then the concentrated solid was dissolved in anhydrous methanol (5 mL). Methyl trifluoroacetate (1.44 g, 11.22 mmol, 1.13 mL) was added to the reaction system, and triethylamine (681.33 mg, 6.73 mmol, 937.17 .Math.L) was added thereto. The reaction mixture was stirred at 38° C. for 12 hours. The reaction mixture was directly concentrated under reduced pressure, and purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound 6-13. [M+1].sup.+ =541.25.

Step 12: Synthesis of Compound 6

[0334] Compound 6-13 (0.1 g, 184.65 .Math.mol) was dissolved in dichloromethane (1 mL), then methyl N-(triethylammoniosulfonyl)carbamate (110.01 mg, 461.63 .Math.mol) was added thereto, and the reaction mixture was stirred at 25° C. for 16 hours, then dichloromethane (10 mL) was added to the reaction mixture, and saturated sodium bicarbonate solution (5 mL) was added thereto for extraction to separate the phases, and the organic phase was added with saturated brine (5 mL) to separate the phases, and then the organic phase was dried over sodium sulfate and concentrated under reduced pressure to obtain a crude product, and the crude product was separated by preparative HPLC (column type: C18 100*30 mm*10 .Math.m; mobile phase: [H.sub.2O (NH.sub.4HCO.sub.3)-ACN]; ACN%: 40%-60%, 8 min) to obtain a crude product. The crude product was separated by SFC analysis method: column type: Chiralpak AD-3, 150x4.6 mm I.D., 3 .Math.m, mobile phase: A: CO.sub.2, B: EtOH (0.1% IPAm, v/v), gradient: time A% B%, 0.0-0.5 min, B% from 10% to 50%, maintained for 4.5 min, 4.5-5.0 min, B% from 50% to 10%, flow rate: 2.5 mL/min, column temperature: 35° C., ABPR: 2000 psi. The retention time of compound 6A was 1.97 minutes, and the retention time of compound 6 was 2.28 minutes. The crude product was separated by SFC (column type: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 .Math.m); mobile phase: [0.1% NH.sub.3H.sub.2O-EtOH]%: 16%-16%, 10 min) to obtain compound 6. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 7.22 - 7.17 (m, 1H), 4.88 - 4.79 (m, 1H), 4.75 - 4.67 (m, 1H), 4.38 -4.33 (m, 1H), 3.53 - 3.32 (m, 2H), 2.93 - 2.84 (m, 1H), 2.71 - 2.58 (m, 1H), 2.33 - 2.05 (m, 3H), 2.04 - 1.74 (m, 16H), 1.72 - 1.60 (m, 2H), 1.57 - 1.41 (m, 3H).

Embodiment 7

[0335] ##STR00541##

Step 1: Synthesis of Compound 7-2

[0336] Compound 7-1 (5 g, 35.16 mmol) was dissolved in tetrahydrofuran (50 mL), and the reaction system was replaced with nitrogen for three times, cooled to 0° C., and a solution of borane in tetrahydrofuran (70.33 mL, 1 M) was slowly added dropwise thereto. The reaction was stirred for 16 hours at 20° C. Sodium hydroxide solution (80 mL, 1 M) was added to the reaction at 0° C., extracted twice with methyl tert-butyl ether (100 mL), and the organic phases were combined and washed with 10% citric acid (80 mLx2) and saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered and concentrated. Compound 7-2 was obtained. .sup.1H NMR (400 MHz, CDC1.sub.3) δ ppm 3.28 (s, 2 H) 1.22 - 1.46 (m, 10 H) 0.85 - 0.87 (m, 3H).

Step 2: Synthesis of Compound 7-3

[0337] Compound 7-2 (4 g, 31.20 mmol) was dissolved in acetonitrile (40 mL), and 2-iodobenzoic acid (13.10 g, 46.80 mmol) was added to the reaction system. The reaction was stirred at 50° C. for 16 hours. The reaction system was filtered and the filtrate was used directly in the next reaction without purification to obtain a solution of compound 7-3 in acetonitrile.

Step 3: Synthesis of Compound 7-4

[0338] The solution of compound 7-3 (4 g, 31.37 mmol) in acetonitrile was dissolved in methanol (60 mL), and the compound R-phenylglycinol (5.22 g, 38.04 mmol) was added to the reaction system, and the reaction mixture was stirred at 20° C. for 2 hours. The reaction system was cooled to 0° C., and trimethylsilyl cyanide (22.01 g, 221.88 mmol) was added thereto, and the reaction mixture was stirred at 50° C. for 16 hours. The reaction mixture was directly evaporated to dryness by rotary evaporation. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound 7-4. [M+1].sup.+ = 273.3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ = 7.42 - 7.29 (m, 5H), 4.17 - 4.01 (m, 1H), 3.99 - 3.68 (m, 2H), 3.58 - 3.39 (m, 1H), 1.60 - 1.16 (m, 10H), 1.10 - 1.04 (m, 3H).

Step 4: Synthesis of Compound 7-5

[0339] Compound 7-4 (5 g, 18.36 mmol) was dissolved in methanol (50 mL) and dichloromethane (50 mL), cooled to 0° C., and lead tetraacetate (13.56 g, 27.53 mmol) was added thereto, and the reaction was stirred at 0° C. for 15 minutes. The reaction mixture was poured into saturated sodium bicarbonate solution (45 mL), extracted three times with dichloromethane (45 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. Compound 7-5 was obtained. [M+1].sup.+ = 241.2.

Step 5: Synthesis of Hydrochloride of Compound 7-6

[0340] Compound 7-5 (3 g, 12.48 mmol) was dissolved in 6 M hydrochloric acid (300 mL), heated to 100° C., and the reaction mixture was stirred for 24 hours. The reaction system was extracted three times with chloroform (300 mL), and the aqueous phase was taken and concentrated under reduced pressure. Hydrochloride of compound 7-6 was obtained. .sup.1H NMR (400 MHz, CD.sub.3OD) δ = 4.47 (s, 1H), 1.66 - 1.42 (m, 10H), 1.22 - 1.14 (m, 3H).

Step 6: Synthesis of Compound 7-7

[0341] Hydrochloride of compound 7-6 (1 g, 5.84 mmol) was dissolved in tetrahydrofuran (10 mL) and water (10 mL), then anhydrous potassium carbonate (2.42 g, 17.52 mmol) and di-tert-butyl dicarbonate (2.55 g, 11.68 mmol) were added thereto, and the reaction was stirred at 20° C. for 16 hours. The pH of reaction mixture was adjusted to 3 with 1 M KHSO.sub.4, and the mixture was extracted four times with dichloromethane (40 mL), then the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 7-7 was obtained. [M-1].sup.- = 270.3.

Step 7: Synthesis of Compound 7-8

[0342] Compound 7-7 (350 mg, 1.29 mmol) was dissolved in N,N-dimethylformamide (4 mL), and 2-(7-azabenzotriazol-1-yl)-N,N,N;N′-tetramethyluronium hexafluorophosphate (735.65 mg, 1.93 mmol) was added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours. N,N-Diisopropylethylamine (833.51 mg, 6.45 mmol) and hydrochloride of compound 1-3 (260.23 mg, 1.68 mmol) were added thereto, then the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (10 mL), extracted twice with ethyl acetate (20 mL), and the organic phases were combined, washed twice with 5% citric acid (15 mL), and washed four times with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (petroleum ether: ethyl acetate=5:1) to obtain compound 7-8. [M+1].sup.+ = 409.4.

Step 8: Synthesis of Compound 7-9

[0343] Compound 7-8 (120 mg, 293.74 .Math.mol) was dissolved in tetrahydrofuran (6 mL), methanol (2 mL) and water (2 mL), then lithium hydroxide monohydrate (36.98 mg, 881.21 .Math.mol) was added thereto and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with 3% citric acid (20 mL), extracted twice with ethyl acetate (20 mL), and the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation to obtain compound 7-9. [M+1].sup.+ = 391.3.

Step 9: Synthesis of Compound 7-10

[0344] Compound 7-9 (30 mg, 76.04 .Math.mol) was dissolved in N,N-dimethylformamide (2 mL), then 1-hydroxybenzotriazole (10.28 mg, 76.04 .Math.mol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (17.49 mg, 91.25 .Math.mol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours. N,N-Diisopropylethylamine (39.31 mg, 304.18 .Math.mol) and hydrochloride of compound BB-1 (19.53 mg, 114.07 .Math.mol) were added thereto, then the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (10 mL), extracted twice with dichloromethane (20 mL), and the organic phases were combined and washed twice with 5% citric acid (15 mL) and washed twice with brine (10 mL), then dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (dichloromethane: methanol =20:1) to obtain compound 7-10. [M+1].sup.+ = 548.4.

Step 10: Synthesis of Trifluoroacetate of Compound 7-11

[0345] Compound 7-10 (150 mg, 273.88 .Math.mol) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (1 mL), and the reaction was stirred at 20° C. for 2 hours. The reaction was directly evaporated to dryness by rotary evaporation by an oil pump, added with dichloromethane and evaporated by rotary evaporation, and the above steps were repeated. Trifluoroacetate of compound 7-11 was obtained.

Step 11: Synthesis of Compound 7-12

[0346] Trifluoroacetate of compound 7-11 (120 mg, 268.11 .Math.mol) was dissolved in methanol (2 mL), then triethylamine (162.78 mg, 1.61 mmol) and methyl trifluoroacetate (343.32 mg, 2.68 mmol) were added thereto. The reaction was heated to 38° C. and stirred for 16 hours. The reaction system was directly concentrated under reduced pressure, then dissolved in water (10 mL) and ethyl acetate (20 mL), and the pH was adjusted to acidic with 3% citric acid (10 mL), then the mixture was extracted for three times with ethyl acetate (20 mL), and then washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 7-12 was obtained. [M+1].sup.+ = 544.3.

Step 12: Synthesis of Compound 7

[0347] Compound 7-12 (130 mg, 239.16 .Math.mol) was dissolved in dichloromethane (4 mL), then tetrahydrofuran (0.4 mL) and Burgess reagent (142.48 mg, 597.89 .Math.mol) were added thereto and the reaction mixture was stirred at 25° C. for 2 hours. The reaction system was added with dichloromethane (10 mL), washed with saturated sodium bicarbonate (5 mL) and then washed with saturated brine (5 mL) to obtain the organic phase, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated by preparative HPLC (column type: C18 100*30 mm*10 .Math.m; mobile phase: [water (NH.sub.4HCO.sub.3)-ACN]; ACN %: 35%-55%, 8 min) to obtain compound 7. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 4.92 - 4.81 (m, 1H), 4.78 - 4.63 (m, 1H), 4.60 (br s, 1H), 3.50 - 3.34 (m, 2H), 2.93 - 2.79 (m, 1H), 2.30 (br s, 2H), 2.10 (br s, 2H), 1.94 - 1.86 (m, 1H), 1.75 (br s, 2H), 1.71 -1.60 (m, 4H), 1.33 (br d, J= 16.5 Hz, 10H), 1.30 - 1.17 (m, 1H), 1.16 - 0.96 (m, 3H).

Embodiment 8

##STR00542##

[0348] Synthetic route:

Step 1: Synthesis of Compound 8-1

[0349] Compound 5-6 (2 g, 7.32 mmol) was dissolved in tetrahydrofuran (20 mL), and the reaction system was replaced with nitrogen for three times, cooled to -70° C., then lithium diisopropylamide (2 M, 7.32 mL) was slowly added dropwise thereto, and the reaction mixture was reacted for 1 hour. Subsequently, at -70° C., methyl iodide (2.60 g, 73.17 mmol, 4.56 mL) was slowly added dropwise, slowly raised to 20° C., and the reaction was continued for 1 hour. Under nitrogen atmosphere, the reaction system was slowly added dropwise with saturated ammonium chloride solution (100 mL) for quenching the reaction, then extracted twice with ethyl acetate (200 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to obtain compound 8-1. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 7.34 - 7.27 (m, 3H), 7.26 - 7.18 (m, 2H), 3.87 - 3.77 (m, 1H), 3.67 - 3.59 (m, 1H), 3.25 (s, 3H), 2.96 (s, 1H), 2.67 - 2.64 (m, 1H), 2.64 - 2.55 (m, 2H), 1.94 (d, J = 10.7 Hz, 1H), 1.43 (d, J= 6.5 Hz, 3H), 1.14 - 1.09 (m, 3H).

Step 2: Synthesis of Compound 8-2

[0350] Compound 8-1 (600 mg, 2.09 mmol) was dissolved in N,N-dimethylformamide (6 mL), then p-toluenesulfonyl hydrazide (466.62 mg, 2.51 mmol) and trifluoromethanesulfonic acid (36.69 mg, 239.58 .Math.mol, 21.58 .Math.L) were added thereto, heated to 100° C., then sodium cyanoborohydride (393.65 mg, 6.26 mmol) was added, and the reaction mixture was reacted for 2 hours. The reaction system was added with ethyl acetate (60 mL), washed with saturated sodium bicarbonate solution (30 mL) and saturated brine (30 mL) in turn to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to obtain compound 8-2. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 7.37 - 7.29 (m, 2H), 7.25 (br d, J = 7.1 Hz, 3H), 3.67 - 3.50 (m, 1H), 3.48 - 3.34 (m, 1H), 3.32 - 3.15 (m, 3H), 2.51 (s, 1H), 2.35 - 2.23 (m, 2H), 2.02 - 1.94 (m, 1H), 1.70 - 1.61 (m, 1H), 1.46 - 1.41 (m, 1H), 1.36 (d, J = 6.5 Hz, 3H), 1.18 - 1.10 (m, 1H), 0.96 (d, J = 7.1 Hz, 3H).

Step 3: Synthesis of Hydrochloride of Compound 8-3

[0351] Compound 8-2 (150 mg, 548.71 .Math.mol) was dissolved in ethanol (4 mL), then 12 M hydrochloric acid (137.18 .Math.L) and wet palladium on carbon (0.5 g, palladium content of 10%) were added thereto, and the reaction mixture was reacted under a hydrogen balloon atmosphere at 15 psi and 20° C. for 16 hours. The reaction system was filtered through diatomite and evaporated to dryness by rotary evaporation to obtain hydrochloride of compound 8-3. [M+1].sup.+=170.1.

Step 4: Synthesis of Compound 8-4

[0352] Compound 1-b (114.81 mg, 496.39 .Math.mol) was dissolved in N,N-dimethylformamide (2 mL), then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (235.93 mg, 620.49 .Math.mol) and diisopropylethylamine (267.31 mg, 2.07 mmol, 360.26 .Math.L) were added thereto, and the reaction mixture was stirred for 30 min, then hydrochloride of compound 8-3 (70 mg) was added thereto, and the reaction mixture was stirred continuously at 20° C. for 2.5 hours. The reaction system was added with ethyl acetate (40 mL), then extracted with 3% citric acid solution (30 mL) and saturated sodium bicarbonate solution (20 mL) in turn to separate the organic phase, and extracted with saturated brine (20 mL) to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 8-4. [M+1].sup.+=383.3.

Step 5: Synthesis of Compound 8-5

[0353] Compound 8-4 (125 mg, 326.80 .Math.mol) was dissolved in a mixed solvent of tetrahydrofuran (3 mL), water (1 mL) and methanol (1 mL), and lithium hydroxide monohydrate (41.14 mg, 980.41 .Math.mol) was added thereto, and the reaction mixture was reacted at 30° C. for 16 hours. The reaction system was added with dichloromethane (40 mL) and 3% citric acid (20 mL) for extraction to separate the organic phase. Then, saturated brine (20 mL) was added for extraction to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 8-5. [M+1].sup.+=369.3.

Step 6: Synthesis of Compound 8-6

[0354] Compound 8-5 (115 mg, 312.10 .Math.mol) was dissolved in N,-dimethylformamide (2 mL), then 1-hydroxybenzotriazole (50.61 mg, 374.53 .Math.mol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (89.75 mg, 468.16 .Math.mol) were added thereto, stirred for 30 min, then diisopropylethylamine (161.35 mg, 1.25 mmol, 217.45 .Math.L) and hydrochloride of BB-1 (64.12 mg, 374.53 .Math.mol) were added thereto and the reaction mixture was reacted at 20° C. for 2 hours. The reaction system was added with ethyl acetate (50 mL) and 3% citric acid (25 mL) for extraction to separate the organic phase, and the organic phase was extracted with saturated brine (25 mL) to separate the organic phase, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduce pressure. The residue was purified by silica gel chromatography (dichloromethane: methanol = 1:0 to 30:1) to obtain compound 8-6. [M+1].sup.+=522.4.

Step 7: Synthesis of Trifluoroacetate of Compound 8-7

[0355] Compound 8-6 (240 mg, 460.08 .Math.mol, 1 eq) was dissolved in dichloromethane (1 mL), then trifluoroacetic acid (0.3 mL) was added thereto, and the reaction mixture was reacted at 20° C. for 1 hour. The reaction system was directly concentrated under reduced pressure to obtain trifluoroacetate of compound 8-7. [M+1].sup.+=422.3.

Step 8: Synthesis of Compound 8-8

[0356] Trifluoroacetate of compound 8-7 (180 mg, 427.01 .Math.mol) was dissolved in methanol (2 mL), then triethylamine (172.84 mg, 1.71 mmol, 237.74 .Math.L) and methyl trifluoroacetate (546.79 mg, 4.27 mmol, 430.54 .Math.L) were added thereto and the reaction mixture was reacted for 3 hours at 38° C. The reaction system was concentrated under reduced pressure, then dissolved with water (10 mL) and ethyl acetate (30 mLx2). The pH was adjusted to acidic with 3% citric acid (5 mL), and the mixture was extracted to separate the organic phase, and the organic phase was washed with saturated brine (15 mL), dried and concentrated to obtain compound 8-8. [M+1].sup.+=518.3.

Step 9: Synthesis of Compound 8

[0357] Compound 8-8 (200 mg, 386.44 .Math.mol) was dissolved in dichloromethane (2 mL), and Burgess reagent (230.23 mg, 966.11 .Math.mol) was added thereto, and the reaction mixture was stirred at 25° C. for 16 hours. .sup.1H NMR (400 MHz, CD.sub.3OD) δ = 5.07 - 4.99 (m, 2H), 4.79 - 4.71 (m, 1H), 4.29 - 4.18 (m, 1H), 3.83 - 3.74 (m, 1H), 3.28 - 3.23 (m, 1H), 2.76 - 2.65 (m, 1H), 2.63 - 2.57 (m, 1H), 2.38 - 2.26 (m, 2H), 2.19 - 2.10 (m, 1H), 2.02 - 1.95 (m, 1H), 1.90 - 1.75 (m, 3H), 1.67 - 1.60 (m, 1H), 1.33 - 1.26 (m, 1H), 1.15 - 1.04 (m, 9H), 0.99 (d, J= 7.1 Hz, 3H). [M+1].sup.+=500.3.

Embodiment 9

[0358] ##STR00543##

[0359] Synthetic route:

Step 1: Synthesis of Compound 9-2

[0360] Compound 9-1 (2 g, 6.15 mmol) was dissolved in methanol (8 mL) and toluene (24 mL), and cooled to 0° C., and a solution of (trimethylsilyl)diazomethane (6.15 mL, 2 M) in n-hexane was slowly added dropwise thereto. The reaction was stirred for 16 hours at 20° C. The reaction mixture was directly evaporated to dryness by rotary evaporation, and then purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound 9-2. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.15 - 5.07 (m, 1H), 3.74 (s, 3H), 2.25 (br s, 1H), 1.72 - 1.41 (m, 23H). [M-99].sup.+ = 240.2.

Step 2: Synthesis of Compound 9-3

[0361] Compound 9-2 (100 mg, 294.62 .Math.mol) was dissolved in dichloromethane (1 mL), cooled to 0° C., and diethylaminosulfurtrifluoride (94.98 mg, 589.23 .Math.mol) was added to the reaction system. The reaction was stirred at 20° C. for 16 hours. The reaction mixture was slowly added to saturated sodium bicarbonate solution (20 mL) at 0° C., and extracted twice with dichloromethane (20 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 9-3. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.20 - 4.99 (m, 1H), 3.75 (s, 3H), 2.32 (br s, 1H), 1.88 - 1.43 (m, 23H). [M-55].sup.+ =286.1.

Step 3: Synthesis of Compound 9-4

[0362] Compound 9-3 (700 mg, 2.05 mmol) was dissolved in tetrahydrofuran (7 mL), water (2.3 mL) and methanol (2.3 mL), then lithium hydroxide monohydrate (430.19 mg, 10.25 mmol) was added thereto and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with water (10 mL) and 5% citric acid (15 mL), extracted twice with ethyl acetate (15 mL), and the organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation. Compound 9-4 was obtained without purification. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.31 (s, 1H), 5.10 (brd, J= 9.5 Hz, 1H), 4.20 - 4.09 (m, 1H), 2.34 (br s, 1H), 1.90 -1.75 (m, 7H), 1.58 (br s, 5H), 1.46 (s, 9H). [M-55].sup.+ = 272.2.

Step 4: Synthesis of Compound 9-5

[0363] Compound 9-4 (776 mg, 2.37 mmol) was dissolved in N,N-dimethylformamide (8 mL), then 1-hydroxybenzotriazole (960.83 mg, 7.11 mmol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (908.77 mg, 4.74 mmol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours. N,N-Diisopropylethylamine (919.02 mg, 7.11 mmol) and hydrochloride of compound 1-3 (367.85 mg, 2.37 mmol) were added thereto, and the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (15 mL), extracted twice with ethyl acetate (30 mL), and the organic phases were combined, washed twice with 5% citric acid (30 mL), and washed four times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (petroleum ether: ethyl acetate=8:1) to obtain compound 9-5. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.32 - 5.21 (m, 1H), 4.49 (br s, 1H), 4.32 (br d, J= 9.8 Hz, 1H), 4.07 (s, 1H), 3.73 (s, 3H), 2.75 (br s, 1H), 2.33 (br s, 1H), 1.89 - 1.53 (m, 18H), 1.44 (s, 9H). [M+1].sup.+ = 465.3.

Step 5: Synthesis of Compound 9-6

[0364] Compound 9-5 (740 mg, 1.59 mmol) was dissolved in tetrahydrofuran (7.4 mL), water (2.47 mL) and methanol (2.47 mL), then lithium hydroxide monohydrate (200.51 mg, 4.78 mmol) was added thereto and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with water (20 mL) and 5% citric acid (25 mL), extracted twice with ethyl acetate (40 mL), and the organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation. Compound 9-6 was obtained without purification. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.30 (brd, J= 9.8 Hz, 1H), 4.48 (brs, 1H), 4.36 (brd, J= 9.9 Hz, 1H), 4.14 (s, 1H), 2.97 (brs, 1H), 2.31 (brs, 1H), 2.01 - 1.50 (m, 19H), 1.44 (s, 9H). [M-55].sup.+ = 395.2.

Step 6: Synthesis of Compound 9-7

[0365] Compound 9-6 (710 mg, 1.58 mmol) was dissolved in 2-butanone (7 mL), then 1-hydroxybenzotriazole (255.52 mg, 1.89 mmol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (453.14 mg, 2.36 mmol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours. N,N-diisopropylethylamine (814.67 mg, 6.30 mmol) and hydrochloride of compound BB-1 (323.74 mg, 1.89 mmol) were added thereto and the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (15 mL), extracted twice with ethyl acetate (30 mL), and the organic phases were combined, washed twice with 5% citric acid (30 mL), and washed four times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (dichloromethane: methanol =20:1) to obtain compound 9-7. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 7.96 (br d, J= 6.3 Hz, 1H), 5.83 - 5.74 (m, 1H), 5.57 - 5.44 (m, 1H), 5.25 (br d, J= 9.9 Hz, 1H), 4.54 - 4.48 (m, 1H), 4.37 (br d, J= 9.9 Hz, 1H), 4.05 - 3.97 (m, 1H), 3.43 - 3.27 (m, 2H), 2.95 - 2.76 (m, 1H), 2.60 - 1.30 (m, 34H). [M+1].sup.+ = 604.4.

Step 7: Synthesis of Trifluoroacetate of Compound 9-8

[0366] Compound 9-7 (760 mg, 1.26 mmol) was dissolved in dichloromethane (7.6 mL) and trifluoroacetic acid (2.7 mL), and the reaction was stirred at 20° C. for 2 hours. The reaction was directly evaporated to dryness by rotary evaporation by an oil pump, added with dichloromethane and evaporated by rotary evaporation, and the above steps were repeated. Trifluoroacetate of compound 9-8 was obtained. [M+1].sup.+ = 504.4.

Step 8: Synthesis of Compound 9-9

[0367] Trifluoroacetate of compound 9-8 (300 mg, 595.70 .Math.mol) was dissolved in methanol (6 mL), then triethylamine (241.12 mg, 2.38 mmol) and methyl trifluoroacetate (762.79 mg, 5.96 mmol) were added thereto. The reaction was heated to 38° C. and stirred for 16 hours. The reaction mixture was dried directly and dissolved by adding water (10 mL) and ethyl acetate (10 mL), and the pH of the solution was adjusted to acidic by adding 5% citric acid (10 mL). The phases were separated, extracted twice with ethyl acetate (10 mL), and the organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 9-9 without purification. [M+1].sup.+ = 600.3.

Step 9: Synthesis of Compound 9

[0368] Compound 9-9 was dissolved in dichloromethane (2.8 mL), and then Burgess reagent (278.20 mg, 1.17 mmol) was added thereto. The reaction was heated to 25° C. and stirred for 2 hours. The reaction mixture was added with sodium bicarbonate solution (10 mL) and saturated brine (5 mL), extracted twice with dichloromethane (15 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by preparative HPLC (column type: C18 100*30 mm* 10 .Math.m; mobile phase: [water (NH.sub.4HCO.sub.3)-ACN]; ACN%: 35%-55%, 8 min) to obtain compound 9. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 9.50 - 8.17 (m, 1H), 7.12- 6.93 (m, 1H), 5.92 - 5.73 (m, 1H), 4.95 - 4.77 (m, 1H), 4.72 - 4.57 (m, 1H), 4.46 (s, 1H), 3.97 - 3.86 (m, 1H), 3.38 (br dd, J= 4.0, 8.9 Hz, 1H), 2.87 - 2.78 (m, 1H), 2.63 - 1.22 (m, 26H). [M+1].sup.+ = 582.3.

Embodiment 10

[0369] ##STR00544##

[0370] Synthetic route:

Step 1: Synthesis of Hydrochloride of Compound 10-1

[0371] Compound 5-5 (3 g, 10.90 mmol) was dissolved in ethanol (80 mL), then hydrochloric acid (1.19 g, 32.69 mmol) and wet palladium on carbon (15 g, 10.68 mmol) were added thereto. The reaction was stirred at 20° C. for 16 hours. The reaction mixture was filtered through diatomite and directly evaporated to dryness by rotary evaporation to obtain a crude product of hydrochloride of compound 10-1. [M+1].sup.+ = 172.0.

Step 2: Synthesis of Compound 10-2

[0372] Compound 1-b (1.87 g, 10.90 mmol) was dissolved in N,N-dimethylformamide (20 mL), then O-(7-azabenzotriazol-1-yl)-N,N,N,N tetramethyluronium hexafluorophosphate (4.78 g, 12.58 mmol) and diisopropylethylamine (4.34 g, 33.55 mmol) were added thereto, after stirring for 30 min, hydrochloride of compound 10-1 (190 mg, 1.12 mmol) was added thereto. The reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (15 mL), extracted twice with ethyl acetate (60 mL), and the organic phases were combined, washed twice with 5% citric acid (30 mL) and washed four times with brine (20 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (petroleum ether: ethyl acetate= 3:1) to obtain compound 10-2. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.28 - 5.16 (m, 1H), 4.50 (br s, 1H), 4.28 (d, J= 9.8 Hz, 1H), 3.92 (s, 1H), 3.74 (s, 3H), 2.81 (s, 1H), 2.67 (s, 1H), 2.17 (br dd, J= 6.1, 12.7 Hz, 1H), 1.99 - 1.93 (m, 1H), 1.90 - 1.84 (m, 1H), 1.59 (brd, J= 13.3 Hz, 2H), 1.43 (s, 9H), 1.04 (s, 9H). [M+1].sup.+ =385.2.

Step 3: Synthesis of Compound 10-3

[0373] Compound 10-2 (500 mg, 1.30 mmol) was dissolved in acetonitrile (7.5 mL), then 2-iodobenzoic acid (976.31 mg, 3.49 mmol) was added thereto, and the reaction mixture was stirred at 60° C. for 16 hours. The reaction mixture was filtered through diatomite and evaporated to dryness by rotary evaporation. Compound 10-3 was obtained without purification. [M-55].sup.+ = 327.1.

Step 4: Synthesis of Compound 10-4

[0374] Compound 10-3 (480 mg, 1.26 mmol) was dissolved in dichloromethane (4.8 mL), cooled to 0° C., and diethylaminosulfur trifluoride (1.01 g, 6.28 mmol) was added thereto, and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was slowly added to saturated sodium bicarbonate solution (20 mL) at 0° C., extracted twice with dichloromethane (20 mL), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 10-4 was obtained without purification. [M-100].sup.+ = 304.0.

Step 5: Synthesis of Compound 10-5

[0375] Compound 10-4 (475 mg, 1.17 mmol) was dissolved in tetrahydrofuran (5.5 mL), water (1.84 mL) and methanol (1.84 mL), then lithium hydroxide monohydrate (147.84 mg, 3.52 mmol) was added thereto and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with water (20 mL) and 5% citric acid (25 mL), extracted twice with ethyl acetate (40 mL), and the organic phases were combined and washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. Compound 10-5 was obtained without purification. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.18 (br d, J= 9.8 Hz, 1H), 4.65 - 4.53 (m, 2H), 4.31 (d, J= 9.9 Hz, 1H), 3.33 (br d, J= 7.3 Hz, 1H), 2.55 - 2.41 (m, 1H), 2.33 - 2.07 (m, 3H), 1.45 (s, 9H), 1.03 (s, 9H). [M-55].sup.+ = 335.1.

Step 6: Synthesis of Compound 10-6

[0376] Compound 10-5 (200 mg, 512.27 .Math.mol) was dissolved in 2-butanone (2 mL), then 1-hydroxybenzotriazole (83.06 mg, 614.72 .Math.mol) and hydrochloride of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (264.83 mg, 2.05 mmol) were added thereto, and the reaction mixture was stirred at 20° C. for 0.5 hours. N,N-Diisopropylethylamine (264.83 mg, 2.05 mmol) and hydrochloride of compound BB-1 (105.24 mg) were added thereto, then the reaction was stirred at 20° C. for 16 hours. The reaction mixture was added with water (15 mL), extracted twice with ethyl acetate (30 mL), and the organic phases were combined, washed twice with 5% citric acid (30 mL) and washed four times with brine (20 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (dichloromethane: methanol =20:1) to obtain compound 10-6. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 8.19 (br d, J= 6.8 Hz, 1H), 5.78 (br s, 1H), 5.49 (br s, 1H), 5.19 (br d, J= 10.0 Hz, 1H), 4.61 (br s, 1H), 4.44 (s, 1H), 4.34 (d, J = 10.3 Hz, 1H), 3.38 (br d, J= 6.5 Hz, 2H), 3.10 (br d, J= 6.5 Hz, 1H), 2.32 - 2.20 (m, 3H), 2.02 - 1.82 (m, 6H), 1.44 (s, 9H), 1.05 (s, 9H). [M+1].sup.+ = 544.3.

Step 7: Synthesis of Trifluoroacetate of Compound 10-7

[0377] Compound 10-6 (180 mg, 331.12 .Math.mol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.7 mL), and the reaction was stirred at 20° C. for 2 hours. The reaction was directly dried by an oil pump, added with a small amount of dichloromethane and dried, and the above steps were repeated until the shape of the product was a light yellow foam. Trifluoroacetate of compound 10-7 was obtained. [M+1].sup.+ = 444.3.

Step 8: Synthesis of Compound 10-8

[0378] Trifluoroacetate of compound 10-7 (145 mg, 326.95 .Math.mol) was dissolved in methanol (3.2 mL), and then triethylamine (132.34 mg, 1.31 mmol) and methyl trifluoroacetate (418.66 mg, 3.27 mmol) were added thereto. The reaction was heated to 38° C. and stirred for 16 hours. The reaction mixture was directly evaporated to dryness by rotary evaporation, dissolved with water (10 mL) and ethyl acetate (10 mL), and the pH of the solution was adjusted to acidic by adding 5% citric acid (10 mL). The phases were separated, extracted twice with ethyl acetate (10 mL), and the organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated to obtain compound 10-8 without purification. [M+1].sup.+ = 540.3.

Step 9: Synthesis of Compound 10

[0379] Compound 10-8 (170 mg, 315.11 .Math.mol) was dissolved in dichloromethane (2.8 mL), and then Burgess reagent (187.73 mg, 787.77 .Math.mol) was added thereto. The reaction was heated to 25° C. and stirred for 2 hours. The reaction mixture was added with sodium bicarbonate solution (10 mL) and saturated brine (5 mL), extracted twice with dichloromethane (15 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by preparative HPLC (column type: C18 100*30 mm*10 .Math.m; mobile phase: (H.sub.2O (NH.sub.4HCO.sub.3)-ACN]; ACN%: 35%-55%, 8 min) to obtain compound 10. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 9.79 - 8.65 (m, 1H), 7.18 - 7.01 (m, 1H), 6.34 - 6.20 (m, 1H), 4.64 - 4.58 (m, 2H), 4.45 - 4.37 (m, 1H), 3.44 - 3.30 (m, 3H), 3.12 - 3.05 (m, 1H), 2.63 - 2.47 (m, 3H), 2.40 - 2.12 (m, 6H), 1.06 (s, 9H). [M+1].sup.+ = 522.3.

Embodiment 11

[0380] ##STR00545##

[0381] Synthetic route:

Step 1: Synthesis of Compound 11-1

[0382] Compound 10-3 (0.7 g, 1.83 mmol) was dissolved in tetrahydrofuran (14 mL), and TEBBE reagent (0.5 M, 14.64 mL) was added thereto at 0° C. The reaction mixture was stirred at 0° C. for 1 hour then heated to 15° C. and stirred continuously for 3 hours. The reaction mixture was slowly poured into saturated sodium bicarbonate solution (50 mL), filtered through diatomite, extracted with ethyl acetate (30 mLx3), and washed with saturated brine (30 mLx2). The residue was purified by column chromatography (petroleum ether: ethyl acetate= 5:1) to obtain compound 11-1. [M+1].sup.+ = 381.1.

Step 2: Synthesis of Compound 11-2

[0383] Under nitrogen atmosphere, diethyl zinc (1 M, 13.14 mL) was slowly added to 1,2-dichloroethane (80 mL) at 0° C. After stirring for 0.25 hours, the reaction mixture was slowly added with diiodomethane (7.04 g, 26.28 mmol, 2.12 mL) at 0° C., and stirred for 0.25 hours. Trifluoroacetic acid (149.84 mg, 1.31 mmol, 97.30 .Math.L) was slowly added to the reaction system, stirred continuously for 0.5 hours. A solution of compound 11-1 (0.5 g, 1.31 mmol) in 1,2-dichloroethane (5 mL) was added to the reaction system, heated to 20° C. and stirred continuously for 12 hours. The reaction was quenched with saturated sodium bicarbonate solution (200 mL), extracted with dichloromethane (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by preparative HPLC (column type: Phenomenex luna C18 80*40 mm*3 .Math.m; mobile phase: [H.sub.2O (HCl)-acetonitrile]; acetonitrile%: 1%-30%,7 min) to obtain compound 11-2. [M+1].sup.+ = 295.2.

Step 3: Synthesis of Compound 11-3

[0384] Compound 11-2 (0.1 g, 339.69 .Math.mol) was dissolved in 1,4-dioxane (3 mL), and then a solution of potassium carbonate (187.79 mg, 1.36 mmol) and di-tert-butyldicarbonate (111.20 mg, 509.53 .Math.mol, 117.06 .Math.L) in water (1 mL) was added thereto, and the reaction was stirred at 15° C. for 12 hours. The reaction mixture was poured into water (30 mL), extracted with ethyl acetate (20 mLx3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 11-3. [M+1].sup.+ = 395.2.

Step 4: Synthesis of Compound 11-4

[0385] Compound 11-3 (88.13 mg, 223.40 .Math.mol) was dissolved in tetrahydrofuran (2 mL) and methanol (0.6 mL), and lithium hydroxide monohydrate (28.12 mg, 670.21 .Math.mol) dissolved in water (0.6 mL) was added thereto. The reaction was stirred at 15° C. for 2 hours. The pH was adjusted to 5 with 3% citric acid, and the mixture was extracted with ethyl acetate (20 mLx3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 11-4. [M+1].sup.+ = 381.3.

Step 5: Synthesis of Compound 11-5

[0386] Compound 11-4 (0.056 g, 148.79 .Math.mol) was dissolved in N,N-dimethylforrnamide (2 mL), then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (84.86 mg, 223.18 .Math.mol) was added to the reaction system, and the reaction was stirred at 15° C. for 0.5 hours. Then diisopropylethylamine (76.92 mg, 595.16 .Math.mol, 103.67 .Math.L) was added to the reaction mixture, and a solution of hydrochloride of compound BB-1 (43.26 mg, 208.31 .Math.mol) dissolved in N,N-dimethylformamide (0.5 mL) was added to the reaction system, and the reaction was stirred at 15° C. for 12 hours. The reaction was diluted with water (20 mL), extracted with ethyl acetate (20 mLx3), and the organic phase was washed with 3% citric acid (20 mL), washed with saturated sodium chloride (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 11-5. [M+1].sup.+ = 534.4.

Step 6: Synthesis of Compound 11-6

[0387] Compound 11-5 (0.02 g, 37.48 .Math.mol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (141.02 mg, 1.24 mmol, 91.57 .Math.L) was added to the reaction system. The reaction mixture was stirred at 15° C. for 1 hour. The reaction was directly quenched with sodium bicarbonate solution (10 mL), extracted with dichloromethane (5 mLx5), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 11-6. [M+1].sup.+ = 434.2.

Step 7: Synthesis of Compound 11

[0388] Compound 11-6 (0.03 g, 69.20 .Math.mol) was dissolved in dichloromethane (1 mL), and trifluoroacetic anhydride (58.13 mg, 276.79 .Math.mol, 38.50 .Math.L) was added to the reaction system. The reaction mixture was stirred at 15° C. for 1 hour. The reaction was directly quenched with sodium bicarbonate solution (10 mL), extracted with dichloromethane (5 mLx5), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by preparative HPLC (column type: Waters Xbridge BEH C18 100*30 mm*10 .Math.m; mobile phase: [H.sub.2O (NHaHCO.sub.3)-acetonitrile]; acetonitrile%: 10%-50%, 8 min) to obtain compound 11. [M+1].sup.+ = 512.2.

Embodiment 12

[0389] ##STR00546##

[0390] Synthetic route:

Step 1: Synthesis of Compound 12-2

[0391] Compound 12-1 (2.36 g, 7.64 mmol) was dissolved in N,N-dimethylformamide (17 mL), then O-(7-azabenzotriazol-1-yl)-N,N,N,N tetramethyluronium hexafluorophosphate (4.36 g, 11.46 mmol) and diisopropylethylamine (3.95 g, 30.55 mmol) were added thereto, after stirring for 30 min, hydrochloride of compound 10-1 (1.7 g, 8.19 mmol) was added thereto, and the reaction was stirred at 15° C. for 1 hour. The reaction mixture was added with water (15 mL), extracted twice with ethyl acetate (60 mL), and the organic phases were combined, washed twice with 5% citric acid (30 mL), and washed four times with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (petroleum ether: ethyl acetate= 1:1) to obtain compound 12-2. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.22 (br d, J=9.76 Hz, 1 H), 4.50 - 4.62 (m, 1 H), 4.09 -4.18 (m, 2 H), 3.74 (br s, 3 H), 1.93 - 2.04 (m, 4 H), 1.87 (br d, J=9.88 Hz, 1H), 1.53 - 1.77 (m, 15 H), 1.39 - 1.46 (m, 9 H), 1.27 (t, J=7.13 Hz, 2 H). [M+1].sup.+ =463.58.

Step 2: Synthesis of Compound 12-3

[0392] Compound 12-2 (3 g, 6.49 mmol) was dissolved in acetonitrile (45 mL), and 2-iodobenzoic acid (3.63 g, 12.97 mmol) was added thereto, and the reaction mixture was reacted at 60° C. for 16 hours. The reaction mixture was filtered through diatomite, and evaporated to dryness by rotary evaporation under reduced pressure. Compound 12-3 was obtained. [M+1].sup.+= 461.56.

Step 3: Synthesis of Compound 12-4

[0393] Compound 12-3 (4 g, 8.69 mmol) was dissolved in tetrahydrofuran (80 mL), cooled to 0° C., and diethylaminosulfurtrifluoride (10.24 g, 34.74 mmol) was added thereto, and the reaction mixture was reacted at 15° C. for 3 hours. The reaction mixture was slowly added to saturated sodium bicarbonate solution (100 mL), and extracted three times with ethyl acetate (50 mL), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 12-4 was obtained. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.227 - 5.251 (br d, J=9.6 Hz, 2 H), 4.862 (br s, 1 H), 4.636 (br s, 1 H), 4.191 - 4.215 (m, 2 H), 3.768 (br s, 3 H), 3.117 (br s, 1 H), 2.369 (br s, 2 H), 2.115 (t, J=14.4 Hz, 1 H), 2.010 - 2.046 (br s, 3 H), 1.618 - 1.701 (m, 13 H), 1.432 (br s, 9 H). [M+1].sup.+ = 459.59.

Step 4: Synthesis of Compound 12-5

[0394] Compound 12-4 (1.90 g, 4.14 mmol) was dissolved in methanol (191 mL), added to another single-necked flask containing a solution of wet palladium on carbon (9.55 g, palladium content of 5%) in methanol, and the reaction system was replaced with a hydrogen balloon for three times, and the reaction mixture was reacted at 15° C. and 15 Psi for 2 hours. The reaction mixture was filtered through diatomite, and evaporated to dryness by rotary evaporation under reduced pressure. Compound 12-5 was obtained. [M+1].sup.+ = 461.60.

Step 5: Synthesis of Compound 12-6

[0395] Compound 12-5 (1.8 g, 3.91 mmol) was dissolved in tetrahydrofuran (40 mL) and methanol (13 mL), then cooled to 0° C., and lithium hydroxide monohydrate (983.94 mg, 23.46 mmol) was added thereto, and the reaction mixture was reacted at 15° C. for 40 hours. The reaction mixture was added with water (30 mL), and the pH was adjusted to 4 to 5 with 5% citric acid (20 mL), then the mixture was extracted twice with ethyl acetate (50 mL), and the organic phases were combined, washed twice with saturated sodium chloride (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. Compound 12-6 was obtained without purification. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 1.424 (br s, 9 H) 1.594 - 1.655 (m, 14 H) 1.846 (br s, 8 H) 2.030 (br s, 1 H) 2.540 - 2.548 (m, 1 H) 3.240 -3.367 (br d, J=50.8 Hz, 1 H) 3.633 - 3.668 (m, 1 H) 3.777 - 3.823 (m, 1 H) 4.446 (br s, 1 H) 6.309 - 6.332 (br d, J=9.2, 1 H). [M+1].sup.+ = 447.58.

Step 6: Synthesis of Compound 12-7

[0396] Compound 12-6 (1.3 g, 2.91 mmol) was dissolved in N,N-dimethylformamide (12 mL), then 1-hydroxybenzotriazole (472.01 mg, 3.49 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (837.07 mg, 4.37 mmol) were added thereto, and the reaction was stirred at 15° C. for 30 min. Hydrochloride of compound BB-1 (747.27 mg, 4.37 mmol) and diisopropylethylamine (1.50 g, 11.64 mmol) were added thereto, and the reaction mixture was reacted at 15° C. for 2 hours. The reaction mixture was added with water (50 mL), extracted twice with ethyl acetate (50 mL), and the organic phases were combined, washed twice with 5% citric acid (30 mL) and washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (dichloromethane: methanol = 20:1) to obtain compound 12-7. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 7.730 - 7.748 (br d, J=7.2 Hz, 1 H), 6.162 (br s, 1 H) 7.270 (br s, 1H), 5.569 (br d, 1H), 5.288 (br s, 1 H), 4.510 - 4.567 (m, 1H), 4.430 (brd, 1H), 4.331 (brd, 1 H), 4.183 - 4.208 (br d, J=10 Hz, 1 H), 3.322 - 3.360 (t, J=7.6 Hz, 2 H), 2.658 - 2.666 (m, 1 H), 2.459 (m, 2 H), 2.017 - 2.076 (m, 1 H), 1.990 (m, 8 H), 1.602 - 1.780 (m, 13 H), 1.424 (m, 9 H), 1.046 (br s, 1 H), 0.976 - 1.029 (br d, J=21.2 Hz, 3 H). [M+1]+ = 600.76.

Step 7: Synthesis of Compound 12-8

[0397] Compound 12-7 (920.00 mg, 1.53 mmol) was dissolved in dichloromethane (18.4 mL), then trifluoroacetic acid (7.89 g, 69.22 mmol) was added thereto, and the reaction mixture was reacted at 15° C. for 1 hour. The reaction mixture was directly evaporated to dryness by rotary evaporation. Compound 12-8 was obtained. [M+1].sup.+= 500.64.

Step 8: Synthesis of Compound 12-9

[0398] Compound 12-8 (420.00 mg, 840.60 .Math.mol) was dissolved in methanol (8.4 mL), then triethylamine (510.36 mg, 5.04 mmol) was added thereto, and methyl trifluoroacetate (1.29 g, 10.09 mmol) was added thereto, and the reaction mixture was heated to 38° C. and stirred for 16 hours. The reaction mixture was concentrated under reduced pressure, added with ethyl acetate (50 mL) and water (20 mL). The pH of the reaction mixture was adjusted to acidic by adding 3% citric acid, and the mixture was extracted three times by adding ethyl acetate (30 mL), and the organic phases were combined, washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (dichloromethane: methanol =10:1) to obtain compound 12-9. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 7.819 - 7.832 (br d, J=5.2 Hz, 1 H), 7.006 - 7.176 (m, 2 H), 5.636 - 5.947 (m, 3 H), 4.343 (br s, 3 H), 3.339 - 3.477 (m, 3 H), 2.666 (br s, 1 H), 2.487 -2.503 (m, 3 H), 2.161 - 2.186 (m, 3 H), 2.000-2.020 (br d, J=8 Hz, 4 H), 1.856 - 1.878 (m, 2H), 1.567 - 1.652 (m, 10 H), 1.003 - 1.061 (m, 5 H). [M+1].sup.+ =596.65.

Step 9: Synthesis of Compound 12

[0399] Compound 12-9 (390.00 mg, 654.74 .Math.mol) was dissolved in dichloromethane (7.8 mL) and tetrahydrofuran (0.78 mL), cooled to 0° C., then Burgess reagent (390.07 mg, 1.64 mmol) was added thereto, and the reaction mixture was reacted at 15° C. for 1 hour. The reaction mixture was added with water (30 mL), extracted twice with ethyl acetate (30 mL), and the organic phases were combined, added with sodium bicarbonate solution (30 mL) and stirred for 20 min, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by preparative HPLC (column type: C18 100*30 mm* 10 .Math.m; mobile phase: [H.sub.2O (NH.sub.4HCO.sub.3)-acetonitrile]; acetonitrile%: 35%-55%, 8 min) to obtain compound 12. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 8.23 (br d, J=7.00 Hz, 1 H), 6.89 - 7.04 (m, 1 H), 5.77 - 5.86 (m, 1 H), 4.94 (br d, J=9.44, 6.91 Hz, 1 H), 4.51 (br d, J=9.26 Hz, 1 H), 4.40 (br s, 1 H), 4.22 - 4.35 (m, 1 H), 3.05 - 3.81 (m, 3 H), 2.71 (br d, J=3.63 Hz, 1 H), 2.39 - 2.66 (m, 2 H), 2.12 - 2.34 (m, 3 H), 1.79 - 2.09 (m, 6 H), 1.61 - 1.73 (m, 11 H), 1.43 (m, 1 H), 1.17 (br d, J=6.88 Hz, 1 H), 0.98 - 1.14 (m, 3 H). [M+1].sup.+ =578.64.

Embodiment 13

[0400] ##STR00547##

[0401] Synthetic route:

Step 1: Synthesis of Compound 13-2

[0402] Compound 13-1 (100.50 mg, 669.25 .Math.mol) was dissolved in N,N-dimethylformamide (25 mL), then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (293.62 mg, 772.21 .Math.mol) and N,N-diisopropylethylamine (266.14 mg, 2.06 mmol, 358.67 .Math.L) were added thereto, and the reaction mixture was stirred for 0.5 hours. Trifluoroacetate of compound 3-5 (0.25 g) was added thereto, and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was extracted by adding with water (15 mL) and ethyl acetate (30 mL× 2), and the organic phases were combined, washed with 5% citric acid (10 mL) and brine (20 mLx4), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (dichloromethane: methanol =5:1) to obtain compound 13-2. [M+1].sup.+ = 618.7.

Step 2: Synthesis of Compound 13

[0403] Compound 13-2 (0.2 g, 323.74 .Math.mol) was dissolved in dichloromethane (6 mL) and tetrahydrofuran (0.6 mL), and Burgess reagent (115.73 mg, 485.61 .Math.mol) was added thereto, and the reaction mixture was reacted at 15° C. for 1 hour. The reaction mixture was washed with saturated sodium bicarbonate (5 mL), extracted with dichloromethane (10 mL), washed with saturated brine solution (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by preparative HPLC (column type: Waters Xbridge Prep OBD C18 150*40 mm* 10 .Math.m; mobile phase: [water (NHaHCO.sub.3)-acetonitrile]; acetonitrile%: 35%-65%, 8 min), and the fraction was concentrated under reduced pressure to obtain compound 13. [M+1].sup.+ = 600.7. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 8.16 (d, J= 7.5 Hz, 1H), 7.18 (s, 4H), 4.98 (q, J= 7.9 Hz, 1H), 4.62 - 4.43 (m, 2H), 3.98 (s, 1H), 3.69 - 3.48 (m, 3H), 3.43 - 3.27 (m, 2H), 2.37 - 2.34 (m, 3H), 1.88 - 1.36 (m, 27H).

Embodiment 14

[0404] ##STR00548##

[0405] Synthetic route:

Step 1: Synthesis of Compound 14-2

[0406] Compound 14-1 (36.44 mg, 214.16 .Math.mol) was dissolved in N,N-dimethylformamide (10 mL), then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (93.96 mg, 247.11 .Math.mol) and N,N-diisopropylethylamine (85.17 mg, 658.95 .Math.mol, 114.78 .Math.L) were added thereto, and the reaction mixture was stirred for 0.5 hours, and trifluoroacetate of compound 3-5 (0.08 g) was added thereto, and the reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (30 mLx 2), and the organic phases were combined, washed with 5% citric acid (10 mL) and brine (20 mLx4), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness by rotary evaporation. The residue was purified by column chromatography (dichloromethane: methanol =5:1) to obtain compound 14-2. [M+1].sup.+ = 638.7.

Step 3: Synthesis of Compound 14

[0407] Compound 14-2 (0.1 g, 156.80 .Math.mol) was dissolved in dichloromethane (3 mL) and tetrahydrofuran (0.3 mL), and Burgess reagent (56.05 mg, 235.21 .Math.mol) was added thereto, and the reaction mixture was reacted at 15° C. for 1 hour. The reaction mixture was washed with saturated sodium bicarbonate (5 mL), extracted with dichloromethane (10 mL), washed with saturated brine solution (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by preparative HPLC (column type: Waters Xbridge Prep OBD C18 150*40 mm* 10 .Math.m; mobile phase: [water (NH.sub.4HCO.sub.3)-acetonitrile]; acetonitrile%: 20%-70%, 8 min), and the fraction was concentrated under reduced pressure to obtain compound 14. [M+1].sup.+ = 620.7. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 8.23 (br d, J= 7.5 Hz, 1H), 7.03 - 7.03 (m, 1H), 7.21 - 6.85 (m, 3H), 5.00 (q, J= 7.8 Hz, 1H), 4.69 - 4.40 (m, 3H), 4.04 - 3.94 (m, 1H), 3.45 - 3.26 (m, 2H), 2.95 - 2.79 (m, 1H), 2.66 - 1.24 (m, 23H).

Embodiment 15

[0408] ##STR00549##

[0409] Synthetic route:

Step 1: Synthesis of Compound 15-1

[0410] At 0° C., N-Boc-L-tert-leucine (0.21 g, 1.10 mmol), N,N-diisopropylethylamine (426.49 mg, 3.30 mmol, 574.79 .Math.L), 2-(7-azobenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (627.38 mg, 1.65 mmol) were added to a solution of hydrochloride of compound 1-3 (305.30 mg, 1.32 mmol) in N,N-dimethylfonnamide (2 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was poured into 5% citric acid solution to separate the phases, and the aqueous phase was extracted with ethyl acetate (20 mL*2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 3:1) to obtain compound 15-1. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.21 (br d,J= 9.6 Hz, 1H), 4.48 - 4.37 (m, 1H), 4.30 - 4.21 (m, 1H), 4.01 - 3.95 (m, 1H), 3.68 - 3.60 (m, 3H), 2.73 - 2.61 (m, 1H), 1.97 - 1.87 (m, 1H), 1.79 - 1.56 (m, 4H), 1.39 - 1.33 (m, 10H), 0.97 (s, 9H).

Step 2: Synthesis of Compound 15-2

[0411] Lithium hydroxide monohydrate (51.25 mg, 1.22 mmol) was added to a solution of compound 15-1 (0.3 g, 814.19 .Math.mol) in tetrahydrofuran (2 mL) and water (1 mL), and the reaction mixture was reacted at 20° C. for 16 hours. The reaction mixture was added with 20 mL of 5% citric acid aqueous solution, and added with 20 mL of ethyl acetate to separate the phases, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. Compound 15-2 was obtained. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.24 (d, J= 9.9 Hz, 1H), 4.51 - 4.45 (m, 1H), 4.41 - 4.33 (m, 1H), 4.18 - 4.15 (m, 1H), 3.06 - 2.99 (m, 1H), 1.99 - 1.89 (m, 1H), 1.85 - 1.75 (m, 3H), 1.59 - 1.49 (m, 2H), 1.46 - 1.42 (m, 9H), 1.05 - 1.01 (m, 9H).

Step 3: Synthesis of Compound 15-3

[0412] At 0° C., hydrochloride of compound BB-1 (196.85 mg, 947.97 .Math.mol), N-methylimidazole (291.87 mg, 3.55 mmol), N,N,N,N-tetramethylchlorofonnamidinium hexafluorophosphate (265.98 mg, 947.97 .Math.mol) were added to a solution of compound 15-2 (0.28 g, 789.98 .Math.mol) in N,N dimethylformamide (3 mL), and reacted at 20° C. for 16 hours. The reaction mixture was poured into 20 mL of water, and a mixed solution of dichloromethane and methanol (volume ratio of 10:1) was added for extraction (20 mL*2), and the organic phase was washed with 5% citric acid solution (20 mL*1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (dichloromethane/methanol = 20:1) to obtain compound 15-3. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 4.71 - 4.23 (m, 3H), 3.97 (br s, 1H), 3.35 (brd, J= 7.1 Hz, 2H), 2.83 - 2.74 (m, 1H), 2.61 - 2.31 (m, 2H), 2.09 (br s, 1H), 2.04 - 1.92 (m, 2H), 1.88 - 1.64 (m, 4H), 1.57 - 1.34 (m, 11H), 1.11 -0.89 (m, 9H).

Step 4: Synthesis of Trifluoroacetate of Compound 15-4

[0413] At 0° C., trifluoroacetic acid (2 mL) was added to a solution of compound 15-3 (0.2 g, 393.99 .Math.mol) in dichloromethane (6 mL), and the reaction mixture was reacted at 20° C. for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain trifluoroacetate of compound 15-4.

Step 5: Synthesis of Compound 15

[0414] At 0° C., pyridine (187.07 mg, 2.37 mmol, 190.89 .Math.L), trifluoroacetic anhydride (206.97 mg, 985.43 .Math.mol, 137.07 .Math.L) were added to a solution of trifluoroacetate of compound 15-4 (0.175 g, 394.17 .Math.mol) in tetrahydrofuran (2 mL), and the reaction mixture was reacted at 20° C. for 4 hours. The reaction mixture was quenched with 20 mL of water, extracted with ethyl acetate (20 mL*2), and the organic phase was washed with 5% citric acid (20 mL*1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by pre-HPLC (column type: Waters Xbridge BEH C18 100*25 mm*5 .Math.m; mobile phase: [H.sub.2O (NHaHCO.sub.3)-ACN]; ACN%: 20%-50%, 10 min) to obtain compound 15. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 9.43 - 8.31 (m, 1H), 7.11 - 6.95 (m, 1H), 5.93 - 5.71 (m, 1H), 4.92 - 4.76 (m, 1H), 4.72 - 4.62 (m, 1H), 4.55 - 4.44 (m, 1H), 4.01 - 3.89 (m, 1H), 3.48 - 3.30 (m, 2H), 2.88 - 2.78 (m, 1H), 2.62 - 2.38 (m, 2H), 2.34 - 2.15 (m, 2H), 2.04 - 1.75 (m, 4H), 1.71 - 1.58 (m, 3H), 1.57 - 1.41 (m, 2H), 1.07 - 0.92 (m, 9H).

Embodiment 16

##STR00550##

[0415] Synthetic route:

##STR00551##

Step 1: Synthesis of Compound 16-2

[0416] Hydrochloride of compound 16-1 (949.93 mg, 4.32 mmol) was added to N,N-dimethylformamide (10 mL), then 2-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (1.97 g, 5.19 mmol) was added thereto and the reaction was stirred for 0.5 hours, then diisopropylethylamine (1.40 g, 10.81 mmol) and N-Boc-L-tert-leucine (1 g, 4.32 mmol) were added thereto, and the reaction was stirred at 20° C. for 16 hours. The reaction mixture was washed with methyl tert-butyl ether (50 mL), water (20 mL), 3% citric acid (20 mL*2) and saturated sodium chloride solution (20 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound 16-2. .sup.1H NMR (400 MHz, CDC1.sub.3) δ = 5.27 - 5.18 (m, 1H), 4.36 (d, J= 4.1 Hz, 1H), 4.33 - 4.28 (m, 1H), 4.23 - 4.13 (m, 2H), 3.89 - 3.80 (m, 1H), 3.79 - 3.70 (m, 1H), 2.77 - 2.61 (m, 2H), 1.98 - 1.82 (m, 2H), 1.80 - 1.70 (m, 1H), 1.69 - 1.60 (m, 2H), 1.54 - 1.47 (m, 1H), 1.46 - 1.41 (m, 9H), 1.29 - 1.25 (m, 3H), 1.06 - 1.00 (m, 9H).

Step 2: Synthesis of Compound 16-3

[0417] Compound 16-2 (0.2 g, 504.39 .Math.mol) was added to tetrahydrofuran (3 mL), and a solution of lithium hydroxide monohydrate (63.50 mg, 1.51 mmol) in water (1.5 mL) was added thereto, and the reaction was stirred at 20° C. for 16 hours. The crude product was neutralized with 3% citric acid solution (20 mL), extracted with ethyl acetate (30 mL), and the organic phase was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Compound 16-3 was obtained without purification. .sup.1H NMR (400 MHz, CD.sub.3OD) δ = 4.32 - 4.25 (m, 2H), 3.90 - 3.81 (m, 2H), 2.85 - 2.67 (m, 2H), 2.01 - 1.86 (m, 2H), 1.79 - 1.50 (m, 5H), 1.44 (s, 9H), 1.06 - 1.00 (m, 9H).

Step 3: Synthesis of Compound 16-4

[0418] Compound 16-3 (0.35 g, 949.88 .Math.mol), hydrochloride of compound BB-1 (197.25 mg, 949.88 .Math.mol) were added to N,N-dimethylformamide (4 mL), cooled to 0° C., then 1-methylimidazole (272.95 mg, 3.32 mmol) and N,N,N,N-tetramethylchloroformamidinium hexafluorophosphate (399.78 mg, 1.42 mmol) were added thereto, and the reaction was gradually warmed to 20° C. and stirred for 16 hours. The reaction mixture was washed with ethyl acetate (50 mL), water (20 mL), 3% citric acid (20 mL*2) and saturated sodium chloride solution (20 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound 16-4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ = 8.15 - 7.87 (m, 1H), 7.24 - 7.14 (m, 1H), 6.19 - 5.95 (m, 1H), 5.81 - 5.55 (m, 1H), 5.44 - 5.24 (m, 1H), 4.51 - 3.72 (m, 5H), 3.49 - 3.27 (m, 2H), 2.86 - 2.64 (m, 2H), 2.55 - 2.26 (m, 2H), 1.97 - 1.75 (m, 5H), 1.73 - 1.55 (m, 3H), 1.51 - 1.36 (m, 10H), 1.06 - 0.90 (m, 9H).

Step 4: Synthesis of Trifluoroacetate of Compound 16-5

[0419] Compound 16-4 (0.31 g, 594.27 .Math.mol) was added to dichloromethane (3 mL), and trifluoroacetic acid (1 mL) was added thereto, and the reaction was stirred at 20° C. for 2 hours. The reaction mixture was concentrated under reduced pressure. Trifluoroacetate of compound 16-5 was obtained. [M+1].sup.+= 422.30

Step 5: Synthesis of Compound 16

[0420] Trifluoroacetate of compound 16-5 (240 mg, 448.13 .Math.mol) was added to tetrahydrofuran (3 mL), cooled to 0° C., then pyridine (212.68 mg, 2.69 mmol) was added thereto, and trifluoroacetic anhydride (235.30 mg, 1.12 mmol) was added dropwise thereto, and the reaction was gradually warmed to 20° C. and stirred for 1 hour. The reaction mixture was washed with ethyl acetate (50 mL), water (10 mL), 3% citric acid (20 mL*2) and saturated sodium chloride solution (10 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol= 20:1) to obtain compound 16. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 8.74 - 8.25 (m, 2H), 7.30 - 7.12 (m, 1H), 4.99 - 4.83 (m, 1H), 4.60 - 4.47 (m, 1H), 4.21 - 4.14 (m, 1H), 3.92 - 3.82 (m, 1H), 3.65 - 3.57 (m, 1H), 3.25 - 3.09 (m, 2H), 2.80 - 2.70 (m, 1H), 2.62 - 2.54 (m, 1H), 2.46 - 2.38 (m, 1H), 2.28 - 2.14 (m, 2H), 1.94 - 1.55 (m, 7H), 1.47 - 1.36 (m, 1H), 1.02 (s, 9H).

Biological Test

Experimental Embodiment 1: Evaluation of the in Vitro Anti-Novel Coronavirus Mpro Protease Activity of the Test Compound

1. Experimental Materials:

1.1 Reagents and Consumables:

[0421] TABLE-US-00001 Names and brands of reagents and consumables Name of reagent and consumable Brand 1 Tris Sigma 2 EDTA Sigma 3 NaCl Sigma 4 384 well plate Perkin Elmer 5 Dimethyl sulfoxide (DMSO) Sigma 6 Substrate (Dabcyl-KTSAVLQSGFRKM-(Edans)) (SEQ ID NO: 1) GenScript 7 SARS-CoV-2 Mpro WuXi AppTec 8 GC376 TargetMol

1.2 Instruments

[0422] TABLE-US-00002 Instruments and brands Instrument Brand 1 SpectraMax M2e microplate reader Molecular Devices 2 Echo 655 liquid workstation Labcyte 3 Tabletop high-speed centrifuge Eppendorf

2. Experimental Method:

[0423] The compound was dissolved in DMSO, and diluted in a 3-fold gradient with Echo655 according to the concentration requirements to 10 concentration points, and duplicate tests were set at each concentration, and the diluted solution was added to a 384-well plate. Mpro protein and substrate were diluted with test buffer (100 mM NaCl, 20 mM Tris-HCl, 1 mM EDTA), and Mpro protein was added to the 384-well test plate, incubated with the compound for 30 min at room temperature, and then the substrate was added thereto, and the test concentration of Mpro protein was 25 nM, and the test concentration of substrate was 25 .Math.M. After incubating for 60 minutes in a 30° C. constant temperature incubator, the fluorescence signal value of Ex/Em=340 nm/490 nm was detected by microplate reader. At the same time, the background well containing the substrate and compound but not containing Mpro protein was detected as control.

3. Data Analysis:

[0424] 1) The inhibition rate was calculated using the following formula: [0426] .sup.#HPE: 100% inhibition control, containing 25 nM Mpro protein + 25 .Math.M substrate + 1 .Math.M GC376 [0427] ZPE: No-inhibition control, containing 25 nM Mpro protein + 25 .Math.M substrate, not containing compound [0428] Compound: Test compound well, containing 25 nM Mpro protein + 25 .Math.M substrate + compound [0429] BG: Background control well, containing 25 .Math.M substrate + compound, not containing Mpro protein [0430] 2) Log (agonist) vs. response -- variable slope nonlinear fitting analysis was carried out on the inhibition rate data (inhibition rate%) of the compound by using GraphPad Prism software, and the IC.sub.50 value of the compound was obtained.

TABLE-US-00003 In vitro anti-novel coronavirus Mpro protease activity of test compounds Compound number IC.sub.50 (nM) 2 249 3 21 4 53 5 35 6 135 7 62 9 25 10 17 11 5.4 12 17 13 75 14 119 15 94 16 43

[0431] Conclusion: The compounds of the present disclosure have good anti-novel coronavirus Mpro protease activity in vitro.

Experimental Embodiment 2: Evaluation of in Vitro Anti-Coronavirus Activity of Compounds by Cytopathic Model

1. Experimental Materials

1.1 Reagents and Consumables

[0432] TABLE-US-00004 Names and brands of reagents and consumables Name of reagent and consumable Brand 1 MEM medium Sigma 2 L-Glutamine Gibco 3 Non-essential amino acid Gibco 4 Double antibody (Penicillin-Streptomycin Solution) HyClone 5 Fetal bovine serum (FBS) ExCell 6 Phosphate buffered saline (DPBS) Corning 7 0.25% Trypsin Gibco 8 CellTiter Glo cell activity assay kit Promega 9 Remdesivir MCE 10 96-well plate Grenier

1.2 Instruments

[0433] TABLE-US-00005 Instruments and brands Instrument Brand 1 Microplate reader BioTek 2 Cell counter Beckman 3 CO.sub.2 incubator Thermo

1.3 Cells and Viruses

[0434] MRC5 cells and coronavirus HCoV OC43 were purchased from ATCC.

[0435] MRC5 cells were cultured in MEM (Sigma) medium supplemented with 10% fetal bovine serum (Excell), 1% double antibody (Hyclone), 1%L-glutamine (Gibco) and 1% non-essential amino acids (Gibco). MEM (Sigma) medium supplemented with 5% fetal bovine serum (Excell), 1% double antibody (Hyclone), 1% L-glutamine (Gibco) and 1% non-essential amino acid (Gibco) was used as the experimental culture medium.

2. Experimental Method

[0436] TABLE-US-00006 Virus test methods used in this study Virus (strain) Cell Compound treatment time (day)/endpoint method Control compound Detection reagent HCoV OC43, 100TCID.sub.50/well 20,000 MRC5 cells/well 5/CPE Remdesivir CellTiter Glo.

[0437] Cells were inoculated into a 96 microwell plate at a certain density (Table 6) and cultured overnight in an incubator at 5% CO.sub.2 and 37° C. On the second day, the compound was added after doubling dilution (8 concentration points, duplicate wells), with 50 .Math.L per well. Then the diluted virus was added to the cells at 100 TCID.sub.50 per well, 50 .Math.L per well. Cell control (cell without compound treatment or virus infection), virus control (cell infected with virus without compound treatment) and culture medium control (only culture medium) were set. The final volume of the culture medium in this experiment was 200 .Math.L, and the final concentration of DMSO in the culture medium was 0.5%. Cells were cultured in a 5% CO.sub.2, 33° C. incubator for 5 days. Cell viability was detected using the cell viability assay kit CellTiter Glo (Promega). Cytotoxicity experiments were performed under the same conditions as antiviral experiments, but without virus infection.

3. Data Analysis:

[0438] The antiviral activity and cytotoxicity of the compound were represented by the inhibition rate (%) and cell viability (%) of the compound on the cytopathic effect caused by the virus at different concentrations, respectively. The calculation formula is as follows:

[00004]$\begin{array}{l}Inhibitionrate\left(\%\right)=\left(readingvalueoftestwell\text{-}\right)\\ \left(averagevalueofviruscontrol\right)/\left(averagevalueofcellcontrol\text{-}\right)\\ \left(averagevalueofviruscontrol\right)\times 100\end{array}$

[00005]$\begin{array}{l}Cellviability\left(\%\right)=\\ \left(\begin{array}{l}readingvalueoftestwell\text{-}\\ averagevalueofculturemediumcontrol\end{array}\right)/\left(\begin{array}{l}averagevalueofcellcontrol\text{-}\\ averagevalueofculturemediumcontrol\end{array}\right)\times 100\end{array}$

[0439] GraphPad Prism was used to perform nonlinear fitting analysis on the inhibition rate and cell viability of the compound, and the half effective concentration (EC.sub.50) and half cytotoxic concentration (CC.sub.50) of the compound were calculated.

TABLE-US-00007 Evaluation of anti-coronavirus activity of compounds in vitro by cytopathic model Compound number EC.sub.50 (nM) CC.sub.50 (nM) 2 697 > 10000 3 62 > 10000 4 141 > 10000 5 205 > 10000 11 3.5 > 10000 15 191 > 10000 16 83 > 10000

[0440] Conclusion: The compounds of the present disclosure have good in vitro anti-coronavirus activity at the cellular level, and have no cytotoxicity.

Embodiment 3: Anti-Novel Coronavirus Activity and Toxicity Test

3.1: Cell Source and Type of New Coronavirus

[0441] Nuclear viruses were obtained from African green monkey kidney (Vero) cells from the American Type Culture Collection (ATCC), Cat. No. CCL-81. Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, WelGene) supplemented with 10% fetal bovine serum (Gibco) and 1% double antibody (Gibco). DMEM medium supplemented with 2% fetal bovine serum (Gibco) and 1% double antibody (Gibco) was used as the experimental culture medium.

[0442] Novel coronavirus βCoV/KOR/KCDC03/2020 strain was provided by Korea Centers for Disease Control and Prevention (KCDC), Serial No. NCCP43326.

3.2: Experimental Process

Cell Plating

[0443] After the Vero cells were digested by trypsin, the Vero cells were diluted to 480,000 cells per mL with experimental culture medium. The diluted cells were added to a 384-well cell test plate with 25 .Math.L and 12,000 cells per well using an automatic liquid separator. Cells were cultured overnight in a 5% CO.sub.2 and 37° C. incubator.

Compound Treatment and Viral Infection

[0444] On the second day, the compound and CP-100356 were diluted with DMSO, and the diluted compound was added to the test cell wells using a liquid workstation. Then, 25 .Math.L of SARS-CoV-2 virus diluted with the experimental culture medium was added to each well, with MOI=0.0125. Cell control (cells without compound treatment or virus infection) and no compound treatment control (cell infected with virus without compound treatment with 0.5% DMSO), and CP-100356 control (cell infected with virus, treated with 2 .Math.M CP-100356) were set. The final volume of cell culture medium in each well was 50 .Math.L. Cells were cultured in a 5% CO.sub.2 and 37° C. incubator for 24 hours.

Immunofluorescence Staining

[0445] (1) After 24 hours of virus infection, 17 .Math.L of 16% paraformaldehyde was added to each well. Then the virus was left at room temperature for 30 minutes; [0446] (2) the supernatant was aspirated and the plate was washed twice with DPBS; [0447] (3) 25 .Math.L of 0.25% Tritonx-100 was added to each well and left at room temperature for 20 minutes; [0448] (4) 0.25% TritonX-100 was aspirated, and DPBS was used to wash the plate twice; [0449] (5) 25 .Math.L of diluted primary antibody (1:3000-fold diluted) was added to each well and incubated at 37° C. for 1 hour; [0450] (6) the primary antibody was aspirated and DPBS was used to wash the plate twice; [0451] (7) 25 .Math.L of diluted secondary antibody Alexa Fluor 488-labeled sheep anti-rabbit IgG (1:2000-fold dilution) and 2.5 .Math.g/mL (1:4000-fold dilution) of Hoechst 33342 were added to each well and incubated for 1 hour at 37° C.; [0452] (8) the secondary antibody and Hoechst were aspirated, and the plate was washed twice with DPBS; [0453] (9) high-content imaging analyzer Operetta was used to read the plate, and the instrument was set as: 488/405 emission, 20 × objective, 5 fields of view per well.

Data Analysis

[0454] Columbus software was used to quantitatively analyze the total number of cells (the number of cells stained by Hoechst) and the number of cells infected by the new coronavirus (the number of cells labeled with Alexa Fluor 488) in the images read by the high-content imaging analyzer. The ratio of infected cells and the total number of cells were used to analyze the antiviral activity and cytotoxicity of the compounds. The calculation formula is as follows:

[00006]$\begin{array}{l}Inhibitionrate\left(\%\right)\text{=}\\ \text{100}\text{-}\left(ratioofinfectedcellsintestwells-\right)\\ \left(averageratioofinfectedcellsincellcontrolwells\right)/\left(averageratioof\right)\\ infectedcellsincontrolwellswithoutcompoundtreatment\text{-}\\ \left(averageratioofinfectedcellsincellcontrolwells\right)\times 100\end{array}$

[00007]$\begin{array}{l}Cellviability\left(\%\right)=totalnumberofcellsintestwells/average\\ totalnumberofcellsincontrolwellswithoutcompound\\ treatment\times 100\end{array}$

[0455] XLfit 4 software was used for nonlinear fitting analysis of the inhibitory activity and cell viability of the compounds, and the IC.sub.50 and CC.sub.50 values of the compounds were calculated. The fitting method was “Sigmoidal dose-response”. The calculation formula of IC.sub.50 and CC.sub.50 is: Y = Bottom + (Top Bottom)/(1 + (IC.sub.50/X)Hillslope).

TABLE-US-00008 Evaluation of anti-coronavirus activity of compounds in vitro by wild-type novel coronavirus Compound number EC.sub.50 (nM) CC.sub.50 (nM) 3 35 > 5000 4 30 > 5000 5 20 > 5000 9 19 > 5000 11 3.5 > 5000

[0456] Conclusion: The compounds of the present disclosure have good anti-novel coronavirus activity in vitro.

Experimental Embodiment 4: Pharmacokinetics Test of Mice

[0457] In this study, C57BL/6J male mice were selected as test animals, and LC/MS/MS method was used to quantitatively measure the plasma concentration of the test compound 11 at different time points after oral administration and injection in mice, so as to evaluate the pharmacokinetic characteristics of the test drug in mice.

[0458] The test compound dissolved in 30% PEG400+70% normal saline was administered to mice (overnight fasting, 6-8 weeks old) by intragastric administration. 25 .Math.L of blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12 and 24 hours after administration, respectively, and placed in commercial anticoagulation tubes pre-added with EDTA-K2, centrifuged at 4° C., 3200 g for 10 min to obtain plasma, and after the plasma sample was treated, the plasma concentration was determined by LC-MS/MS method.

TABLE-US-00009 Pharmacokinetic parameters of compound 11 in mice PK parameter i.v.@ 3mpk p.o.@ 10 mpk C.sub.max (nM) NA 1855 T.sub.max (h) NA 0.25 T.sub.½ (h) 0.2 NA Vd.sub.ss (L/kg) 0.9 NA CL(mL/min/kg) 68.5 NA AUC.sub.0-last (nM.Math.h) 1447 1519

[0459] NA means not present;

[0460] Conclusion: The compounds of the present disclosure are cleared quickly in mice, resulting in low exposure of the compounds, and about 30% bioavailability for oral absorption.

Experimental Embodiment 5: Pharmacokinetic Test of the Compound of the Present Disclosure in Combination with Ritonavir

[0461] In this study, C57BL/6J male mice were selected as test animals, and the LC/MS/MS method was used to quantitatively measure the plasma concentrations of mice in combination with Ritonavir at different time points, so as to evaluate the pharmacokinetic characteristics of the test drug in mice.

[0462] Firstly, 10 mpk of Ritonavir was administered to mice by intragastric administration at -12 h and 0 h, and then the test compound was dissolved in 30% PEG400+70% normal saline solution and administered to mice by intragastric administration (overnight fasting, 6 to 8 weeks old). 25 .Math.L of blood was collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12 and 24 hours after administration to animals, placed in a commercial anticoagulant tube pre-added with EDTA-K2, and centrifuged at 4° C., 3200 g for 10 min to obtain plasma. After the plasma samples were treated, the plasma concentration was determined by LC-MS/MS method.

TABLE-US-00010 Pharmacokinetic parameters of compound 11 in combination with Ritonavir in mice PK parameter p.o.@ 10 mpk C.sub.max (nM) 10090 T.sub.max (h) 0.5 AUC.sub.0-last (nM•h) 28683

[0463] Conclusion: After the compound of the present disclosure is used in combination with Ritonavir, the exposure is increased by nearly 20 times compared with single drug.

Experimental Embodiment 6: Tissue Distribution Test in Rats

[0464] In this study, SD male rats were selected as test animals, and the drug concentrations of test compounds in plasma and lungs of rats at different time points were quantitatively measured by LC/MS/MS method to evaluate the pharmacokinetic characteristics of test drugs in rats.

[0465] Firstly, 10 mpk of Ritonavir was administered to rats by intragastric administration at -12 h and 0 h, and 30 mpk of the test compound was dissolved in 10% Solutol + 30% PEG 400 + 2% Tween 80+H.sub.2O 58% solution and administered to rats by intragastric administration (overnight fasting). 40 .Math.L of blood was collected from the saphenous vein of rats at 0.25, 1 and 6 hours after administration, placed in an anticoagulant tube added with EDTA-K2, centrifuged at 4° C., 3200 g for 10 min to obtain plasma, and some animals were killed at 0.25, 1 and 6 hours respectively to collect lung tissue. After the plasma samples were treated, the plasma concentration was determined by LC-MS/MS method.

TABLE-US-00011 Pharmacokinetic parameters of compound 11 in rats Tissue concentration and ratio at each time point Lung/plasma (nM) Ratio 0.25 h 15800/2860 5.5/1 1h 18450/5490 3.2/1 6h 7850/4875 1.6/1 AUC(0-6 h) (h.nmol/L) 76840/29371 2.6/1

[0466] Conclusion: After the compound of the present disclosure is used in combination with Ritonavir, there is higher exposure in the lungs in rats.