PHARMACEUTICAL USE OF ALDEHYDE-BASED COMPOUND
20230138310 · 2023-05-04
Inventors
- Hong Liu (Shanghai, CN)
- Jian Li (Shanghai, CN)
- Wenhao DAI (Shanghai, CN)
- Jingjing PENG (Shanghai, CN)
- Xiong XIE (Shanghai, CN)
- Shulei HU (Shanghai, CN)
- Chunpu LI (Shanghai, CN)
- Yechun XU (Shanghai, CN)
- Haitao Yang (Shanghai, CN)
- Leike ZHANG (Shanghai, CN)
- Haixia SU (Shanghai, CN)
- Hualiang Jiang (Shanghai, CN)
- Zhenming Jin (Shanghai, CN)
- Gengfu XIAO (Shanghai, CN)
- Kaixian Chen (Shanghai, CN)
Cpc classification
C07D409/12
CHEMISTRY; METALLURGY
C07D411/12
CHEMISTRY; METALLURGY
A61P29/00
HUMAN NECESSITIES
A61K31/4025
HUMAN NECESSITIES
A61K31/4184
HUMAN NECESSITIES
C07D207/27
CHEMISTRY; METALLURGY
A61K31/403
HUMAN NECESSITIES
A61K31/498
HUMAN NECESSITIES
A61K31/416
HUMAN NECESSITIES
A61K31/4439
HUMAN NECESSITIES
A61K31/4709
HUMAN NECESSITIES
A61K31/40
HUMAN NECESSITIES
C07D405/12
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
A61K31/437
HUMAN NECESSITIES
A61K31/422
HUMAN NECESSITIES
C07D401/12
CHEMISTRY; METALLURGY
A61K31/4155
HUMAN NECESSITIES
A61P1/00
HUMAN NECESSITIES
International classification
C07D403/12
CHEMISTRY; METALLURGY
C07D405/12
CHEMISTRY; METALLURGY
C07D413/12
CHEMISTRY; METALLURGY
C07D401/12
CHEMISTRY; METALLURGY
C07D411/12
CHEMISTRY; METALLURGY
Abstract
Use of an aldehyde-based compound as represented by general formula I, a pharmaceutical composition, a pharmaceutical salt, an enantiomer, a diastereomer and a racemic compound thereof as a novel coronavirus 2019 (2019-nCov) 3CL protease inhibitor in preparation of a medicament for treating and/or preventing and relieving respiratory tract infections, pneumonia and other related diseases caused by the novel coronavirus infection 2019.
##STR00001##
Claims
1. A method for treatment, prevention and/or alleviation of a related disease caused by 2019-nCov infection, which comprising the step: administrating aldehyde compound according to General Formula I, or a pharmaceutically acceptable salts, enantiomer, diastereomer or racemate thereof to a subject in need thereof: ##STR00184## wherein, the chiral carbon atoms C*, C*.sup.2, C*.sup.3, and C*.sup.4 are each independently in S configuration, R configuration, or the combinations thereof; n=0 or 1; R.sub.1 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C3-C7 cycloalkyl, trifluoromethyl, C2-C6 alkynyl, 4-7 membered heterocyclyl, C5-C7 aryl, and 5-7 membered heteroaryl; wherein each of the heterocyclyl and heteroaryl contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; and the substituents are each independently selected from the groups consisting of halogen, C1-C4 straight or branched alkyl, C1-C4 straight or branched alkenyl, C2-C4 straight or branched alkynyl, C1-C4 straight or branched alkoxy, C1-C4 straight or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C1-C4 acyl, amide, sulfonyl, aminosulfonyl, and C1-C4 alkyl-substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are connected form 5-7 membered ring; R.sub.2 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C3-C7 cycloalkyl, 5-12 membered heterocyclyl, C6-C12 aryl, 5-12 membered heteroaryl, styryl, and -Cbz; wherein each of the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; the substituents are each independently selected from halogen, C1-C6 straight or branched alkyl, C2-C6 straight or branched alkenyl, C2-C6 straight or branched alkynyl, C1-C6 straight or branched alkoxy, C1-C6 straight or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C1-C4 acyl, amide, sulfonyl, aminosulfonyl, and C1-C4 alkyl-substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are connected form 5-7 membered ring; R.sub.3 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C1-C6 straight or branched alkyl, C1-C6 straight or branched alkoxy, C3-C7 cycloalkyl, C6-C12 aryl, and 5-12 membered heteroaryl, wherein the heteroaryl contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; wherein, the substituents are each independently selected from the group consisting of halogen, C1-C6 straight or branched alkyl, C2-C6 straight or branched alkenyl, C2-C6 straight or branched alkynyl, C1-C6 straight or branched alkoxy, C1-C6 straight or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C1-C4 acyl, amide, sulfonyl, aminosulfonyl, and C1-C4 alkyl-substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are connected form 5-7 membered ring.
2. The method of claim 1, wherein the related diseases caused by 2019 n-Cov infection is selected from the groups consisting of respiratory tract infection, pneumonia and complications thereof, or the combinations thereof.
3. The method of claim 1, wherein R.sub.1 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of trifluoromethyl, alkynyl, cyclopropanyl, cyclobutanyl, cyclopentyl, cyclohexyl, phenyl, thienyl, pyrazolyl, thiazolyl, pyridyl, and furyl.
4. The method of claim 1, wherein R.sub.2 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of phenyl, styryl, benzoheterocyclyl, and 5-12 membered heteroaryl.
5. The method of claim 1, wherein the chiral carbon atoms C*, C*.sup.2, C*.sup.3, and C*.sup.4 are in S configuration R.sub.3 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C1-C6 straight or branched alkyl, C3-C7 cycloalkyl, and phenyl.
6. The method of claim 1, wherein the compounds in General formula I is selected from the groups consisting of: TABLE-US-00005 No. Name Structure 1 6-Chloro-N-{(S)-3-methyl-1-carbonyl-1- {{(S)-1-carbonyl-1-{{(S)-1-carbonyl-3- [(S)-2-carbonylpyrrolidine-3-yl]propan-2-yl} amino}-3-phenylpropan-2-yl}amino}butan- 2-yl}-2H-chromene-3-carboxamide
7.-9. (canceled)
10. An aldehyde compound according to General Formula I, or a pharmaceutically acceptable salts, enantiomers, diastereomers or racemates thereof; ##STR00273## wherein, the chiral carbon atoms C*, C*.sup.2, C*.sup.3, and C*.sup.4 are each independently in S configuration, in R configuration, or the combinations thereof; n=1; R.sub.1 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C3-C7 cycloalkyl, trifluoromethyl, C2-C6 alkynyl, 4-7 membered heterocyclyl, and 5-7 membered heteroaryl; wherein each of the heterocyclyl and heteroaryl contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; the substituents are each independently selected from the groups consisting of halogen, C1-C4 straight or branched alkyl, C1-C4 straight or branched alkenyl, C2-C4 straight or branched alkynyl, C1-C4 straight or branched alkoxy, C1-C4 straight or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C1-C4 acyl, amide, sulfonyl, aminosulfonyl, and C1-C4 alkyl-substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are connected form 5-7 membered ring; R.sub.2 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C3-C7 cycloalkyl, 5-12 membered heterocyclyl, 6-membered aryl fused 5-7 membered heterocyclyl), C6-C12 aryl, 5-12 membered heteroaryl, styryl, and -Cbz; wherein each of the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; the substituents are each independently selected from halogen, C1-C6 straight or branched alkyl, C2-C6 straight or branched alkenyl, C2-C6 straight or branched alkynyl, C1-C6 straight or branched alkoxy, C1-C6 straight or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C1-C4 acyl, amide, sulfonyl, aminosulfonyl, and C1-C4 alkyl-substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are connected form 5-7 membered ring; R.sub.3 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C1-C6 straight or branched alkyl, C1-C6 straight or branched alkoxy, C3-C7 cycloalkyl, C6-C12 aryl, and 5-12 membered heteroaryl, wherein the heteroaryl contains 1-3 heteroatoms selected from oxygen, sulfur and nitrogen; wherein, the substituents are each independently selected from halogen, C1-C6 straight or branched alkyl, C2-C6 straight or branched alkenyl, C2-C6 straight or branched alkynyl, C1-C6 straight or branched alkoxy, C1-C6 straight or branched alkylcarbonyloxy, cyano, nitro, hydroxyl, amino, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C1-C4 acyl, amide, sulfonyl, aminosulfonyl, and C1-C4 alkyl-substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are connected form 5-7 membered ring.
11. The compound of claim 10, wherein R.sub.1 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of trifluoromethyl, alkynyl, cyclopropanyl, cyclobutanyl, cyclopentyl, cyclohexyl, phenyl, thienyl, pyrazolyl, thiazolyl, pyridyl, and furyl.
12. The compound of claim 10, wherein R.sub.2 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of phenyl, styryl, benzoheterocyclyl, and 5-12 membered heteroaryl.
13. The compound of claim 10, wherein the chiral carbon atoms C*, C*.sup.2, C*.sup.3, and C*.sup.4 are in S configuration, R.sub.3 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of C1-C6 straight or branched alkyl, C3-C7 cycloalkyl, and phenyl.
14. The compound of claim 10, wherein the compounds in General Formula I is selected from the group consisting of: TABLE-US-00006 No. Name Structure 1 6-Chloro-N-{(S)-3-methyl-1-carbonyl-1- {{(S)-1-carbonyl-1-{{(S)-1-carbonyl-3- [(S)-2-carbonylpyrrolidine-3-yl]propan-2- yl}amino}-3-phenylpropan-2-yl}amino}butan- 2-yl}-2H-chromene-3-carboxamide
15. (canceled)
16. The compound of claim 10, wherein R.sub.2 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of phenyl, styryl, benzodioxole, indole, isoxazole, 2-hydroproppyran, pyridine, pyrazole, dihydroimidazopyridine, imidazopyridine, benzothiophene, dihydrobenzodioxane, quinoxaline, benzofuran, indazole, benzimidazole, and quinoline.
17. An aldehyde compound according to General Formula I, or a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof; wherein the compound is: ##STR00298##
18. The method of claim 1, wherein R.sub.2 is a group unsubstituted or substituted with 1-3 substituents, and the group is selected from the group consisting of phenyl, styryl, benzodioxole, indole, isoxazole, 2-hydroproppyran, pyridine, pyrazole, dihydroimidazopyridine, imidazopyridine, benzothiophene, dihydrobenzodioxane, quinoxaline, benzofuran, indazole, benzimidazole, and quinoline.
19. A pharmaceutical composition, wherein the pharmaceutical composition comprises (a) the aldehyde compound of claim 10, or the pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof; and (b) a pharmaceutically acceptable carrier.
Description
DESCRIPTION OF THE DRAWINGS
[0051]
[0052]
[0053]
EMBODIMENTS FOR CARRYING OUT THE INVENTION
[0054] After extensive and in-depth research and extensive screening, the inventors first time unexpectedly developed a class of active ingredient that can effectively inhibit the 2019-nCov, that is, the compounds according to Formula I, or pharmaceutically acceptable salts, enantiomers, diastereomers or racemates thereof. Tests have shown that the active ingredient of the present invention can effectively inhibit the 3CL protease activity of 2019-nCov, thereby inhibiting the replication and viability of 2019-nCov. The present invention has been completed on this basis.
[0055] Terms
[0056] Unless otherwise specified, the term “substituted” herein refers to the substitution of one or more hydrogen atoms on the group with substituents selected from the group consisting of C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.10 cycloalkyl, C.sub.1-C.sub.10 alkoxy, halogen, hydroxyl, carboxyl (—COOH), C.sub.1-C.sub.10 aldehyde group, C.sub.2-C.sub.10 acyl, C.sub.2-C.sub.10 ester group, amino, phenyl; the phenyl includes unsubstituted or substituted phenyl with 1-3 substituents, the substituents are selected from halogen, C.sub.1-C.sub.10 alkyl, cyano, OH, nitro, C.sub.3-C.sub.10 cycloalkyl, C.sub.1-C.sub.10 alkoxy, and amino.
[0057] Unless otherwise specified, each chiral carbon atom in all the compounds herein may optionally be in the R configuration or the S configuration, or mixtures of the R configuration and the S configuration.
[0058] The term “C.sub.1-C.sub.6 alkyl” refers to straight or branched chain alkyl groups having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or similar groups.
[0059] The term “3-8 membered heterocyclyl” refers to groups formed by losing one hydrogen atom of 3-8 membered saturated rings with 1-3 heteroatoms selected from the group consisting of N, S, O; for example, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, or similar groups.
[0060] The term “6-10 membered aryl” refers to groups formed by losing one hydrogen atom of 6-10 membered aryl groups; for example, phenyl group, naphthyl group, or similar groups.
[0061] The term “5-10 membered heteroaryl” refers to groups formed by losing one hydrogen atom of 5-8 membered aryl groups h with 1-3 heteroatoms selected from the group consisting of N, S, O, where each ring system of heteroaryl can be monocyclic or polycyclic; for example, pyrrolyl, pyridyl, thienyl, furyl, imidazolyl, pyrimidinyl, benzothienyl, indolyl, imidazopyridyl, quinolinyl, or similar groups.
[0062] The term “C1-C6 alkoxy” refers to straight or branched alkoxy groups having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy, tert-butoxy, or similar groups.
[0063] The term “C2-C6 ester group” refers to R—O—C(═O)— groups having 2-6 carbon atoms, such as —COOCH.sub.3, —COOC.sub.2H.sub.5, —COOC.sub.3H.sub.7, —COOC.sub.4H.sub.9, or similar groups.
[0064] The term “C2-C6 alkenyl” refers to groups formed by losing one or two hydrogen atoms of olefins with 2-6 carbon atoms. The olefin may be monoolefin, diolefin or triolefin, for example, —CH═CH.sub.2, —C.sub.2H.sub.4═CH.sub.2, —CH═C.sub.2H.sub.4, or similar groups.
[0065] The term “halogen” refers to F, Cl, Br, and I.
[0066] Unless otherwise specified, the structural formula described herein are intended to include all isomeric forms (such as enantiomeric, diastereomeric, and geometric isomers (or conformational isomers)): for example, R, S configuration of asymmetrical centers, (Z), (E) isomers of double bonds, and (Z), (E) conformational isomers, etc. Therefore, the single stereochemical isomers or enantiomers, diastereomers or geometric isomers (or conformers) of the compounds of the invention, or mixtures thereof all fall within the scope of the invention.
[0067] The term “tautomer” means that structural isomers having different energies can exceed the low energy barrier and thereby transform between each other. For example, proton tautomers (proton shift) includes interconversion by proton transfer, such as 1H-indazole and 2H-indazole, 1H-benzo[d]imidazole and 3H-benzo[d]imidazole. Valence tautomers include interconversion through some bonding electron recombination.
[0068] The form “C1-C6” herein means that the group can have 1 to 6 carbon atoms, such as 1, 2, 3, 4, or 5.
[0069] Active Ingredient
[0070] In the present invention, an active ingredient that can effectively inhibit the replication of the 2019-nCov is provided. The active ingredient is the compound r according to General Formula I, and the active ingredient can effectively prevent, treat and/or alleviate 2019-nCov related diseases.
[0071] Tests have shown that the active ingredient of the present invention can effectively inhibit the 3CL protease of the 2019-nCov, thereby inhibiting the replication of the 2019-nCov, thereby preventing, treating and/or alleviating 2019-nCov related diseases. 2019-nCov is also known as SARS-CoV-2.
[0072] It should be understood that, the active ingredient of the present invention includes the aldehyde compounds according to Formula (I), or pharmaceutically acceptable salts, enantiomers, diastereomers, or racemates thereof, or prodrugs thereof. It should be understood that, the active ingredient of the present invention also includes the crystalline forms, amorphous compounds, and deuterated compounds of the compound of Formula (I).
[0073] The “pharmaceutically acceptable salt” refers to the conventional non-toxic salts formed by reacting the compounds of Formula (I) with inorganic acids or organic acids. For example, the conventional non-toxic salts can be prepared by reacting the compounds of Formula (I) with inorganic acids or organic acids, the inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid and phosphoric acid, etc, organic acids include citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalene disulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid and isethionic acid, etc.; or the sodium, potassium, calcium, aluminum or ammonium salts formed with inorganic bases after the compounds of Formula (I) formed esters with propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, aspartic acid, or glutamic acid; or amine, ethylamine or ethanolamine salts formed by the compounds of Formula (I) with organic bases; or the corresponding inorganic acid salts formed with hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid or phosphoric acid after the compounds of Formula (I) formed esters with lysine, arginine, ornithine or the corresponding organic acid salts formed with formic acid, acetic acid, picric acid, methanesulfonic acid or ethanesulfonic acid.
[0074] Pharmaceutical Composition and Use
[0075] The present invention also provides the use of the mixtures of one or more of the aldehyde compounds according to Formula (I), or pharmaceutically acceptable salts, enantiomers, diastereomers or racemates and prodrugs thereof as active ingredients for the preparation of drugs for the treatment and/or prevention, alleviation of respiratory infections, pneumonia and other related diseases caused by 2019 n-Cov infection.
[0076] The pharmaceutical composition provided by the present invention preferably contains the active ingredient in a weight ratio of 0.001-99 wt %, the preferred ratio is that the compound of Formula I as the active ingredient accounts for 0.1 wt %-90 wt % of the total weight, and the rest is pharmaceutically acceptable carrier, diluent, solution or salt solution.
[0077] When needed, one or more pharmaceutically acceptable carriers can be added to the drug. The carriers include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants and the like in the pharmaceutical field.
[0078] The compounds and pharmaceutical compositions provided by the present invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, etc., and can be present in suitable solid or liquid carriers or diluents, also in the sterilization equipments for injection or drip infusion.
[0079] Various dosage forms of the pharmaceutical compositions of the present invention can be prepared according to conventional preparation methods in the pharmaceutical field. The unit measurement of the preparation formula usually contains 0.05-400 mg of the compounds of Formula I, preferably, the unit measurement of the preparation formula contains 1 mg-500 mg of the compounds of Formula I.
[0080] The compounds and pharmaceutical compositions of the present invention can be used on mammals in clinical usage, including humans and animals, and can be administered via pathways such as oral, nose, skin, lung, or gastrointestinal. Most preferred is oral administration. The most preferred daily dose is 0.01-400 mg/kg body weight, taken at one time, or 0.01-200 mg/kg body weight, taken in divided doses. Regardless of the administration method, the individual's optimal dosage should be determined based on the specific treatment. Usually it should start with smaller dose and increase the dose gradually until the most suitable dose is found. The drugs or inhibitors of the present invention can be administered in various ways, for example, it can be introduced into the body such as muscle, intradermal, subcutaneous, vein, mucosal tissue by methods such as injection, spraying, nose drops, eye drops, penetration, absorption, physical or chemically mediation; or be introduced into the body by mixed or wrapped by other substances.
[0081] The main advantages of the present invention include:
[0082] (a) The compounds of the present invention can effectively inhibit 2019-nCoV 3CL protease, and some of the compounds have IC.sub.50 values at about 70 nM.
[0083] (b) The compounds of the present invention has a higher inhibitory rate on 2019-nCoV at the viral level than the positive control CQ, showing a better anti-2019-nCoV potentiality.
[0084] (c) The compounds of the present invention have low toxic and side effects and good drug-making properties.
[0085] The present invention will be further illustrated below with reference to the specific examples. It should be understood that these examples are only to illustrate the invention but not to limit the scope of the invention. The experimental methods with no specific conditions described in the following examples are generally performed under the conventional conditions, or according to the manufacturer's instructions. Unless indicated otherwise, parts and percentage are calculated by weight.
[0086] The analytical data of samples were measured by the following instruments: NMR was measured by GEMINI-300, Bruker AMX-400 and INVOA-600 NMR instruments, TMS (tetramethylsilane) as the internal standard, the unit of chemical shift was ppm, the unit of coupling constant was Hz; the mass spectrum was measured by Finnigan MAT-711, MAT-95 and LCQ-DECA mass spectrometers and IonSpec 4.7 Tesla mass spectrometer. 200-300 Mesh silica gel (produced by Qingdao Ocean Chemical Plant) was used for column chromatography; TLC silica gel plate was HSGF-254 thin-layer chromatography prefabricated plate produced by Yantai Chemical Plant; boiling range of petroleum ether was 60-90° C.; UV lamp and iodine cylinder was used for color development. Unless otherwise specified, the conventional reagents and medicines used in the following examples were purchased from Sinopharm Group. The reagents and solvents used in the experiment were handled according to the specific conditions of the reaction.
Example 1: Synthesis of Compound 1
[0087] ##STR00092##
synthetic route:
##STR00093## ##STR00094##
Synthesis of Compound 1-2
[0088] Under argon protection, dimethyl N-tert-butoxycarbonyl-L-glutamate 1-1 (6 g, 21.8 mmol) was dissolved in 60 mL of anhydrous tetrahydrofuran, and LiHMDS (1M in THF) in tetrahydrofuran solution (47 mL, 47 mmol) was slowly added dropwise at −78° C., while the temperature was kept stable at −78° C. during the process for about 1 hour. The mixture was stirred at −78° C. for 1 hour. Bromoacetonitrile (2.79 g, 23.3 mmol) was dissolved in 20 ml of tetrahydrofuran, and then the solution was slowly added dropwise into the system for 1-2 hours. The temperature was controlled at −78° C., and the reaction was continued for 20 hours. After THL monitoring (alkaline potassium permanganate was used for color development) showed that the reaction was completed, 3 mL of methanol and 22.7 mL (v/v=1/7.5) of the mixture of glacial acetic acid and tetrahydrofuran were added to the reaction solution to quench. The mixture was warmed to room temperature after stirred for 10 min. 40 mL of saturated sodium chloride solution was poured and well stirred. The reaction system was observed to have layered. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (EA). The combined organic layer was dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography (PE:EA=4:1) to obtain a pale yellow oily substance 1-2 3.9 g, yield 58%.
Synthesis of Compound 1-3
[0089] Compound 1-2 (1 g, 3.15 mmol) was dissolved in 25 mL of anhydrous methanol, after stirred under an ice bath to 0° C., cobalt dichloride hexahydrate (450 mg, 1.89 mmol) was added and the solution turned to purple-red. 10 minutes later, sodium borohydride (715 mg, 18.9 mmol) was added in small portions, and the color of the solution turned to purple-black. The reaction solution continued to react in an ice bath for 1 hour and then warmed to room temperature. After 15 h, the mixture was quenched with 5 mL of saturated NH.sub.4Cl solution and stirred for 10 min. The solid was filtered off with diatomite, and the filtrate was evaporated to dryness under reduced pressure, anextracted with 20 mL of water and 30×3 mL of ethyl acetate. The combined organic phase was dried with anhydrous Na.sub.2SO.sub.4 for 1 h, then concentrated under reduced pressure and separated by column chromatography [PE:EA=1:2] to obtain white powdery solid Compound 1-3 460 mg, yield 51%.
Synthesis of Compound 1-4
[0090] Intermediate 1-3 (1 g, 3.5 mmol) was dissolved in dichloromethane, 4 M HCl in dioxane solution (9 mL, 35 mmol) was added at 0° C., and the reaction was stirred at room temperature for 12 hours. The solution was evaporated to dryness to obtain Intermediate 1-4 and used directly to the next reaction.
Synthesis of Compound 1-6
[0091] Compound 1-5 (1.1 g, 3.5 mmol) was dissolved in dichloromethane (40 mL), the reaction solution was cooled to −20° C., then HATU (1.9 g, 4.9 mmol) was added to the reaction solution. After stirred for twenty minutes, Intermediate 1-4 obtained in the previous step was added to the reaction solution and stirred at −20° C. for 30 minutes. DIPEA (1.7 mL, 10.5 mmol) was added dropwise to the reaction solution. After stirred for 12 hours, the reaction was extracted with ammonium chloride (40×3 mL), sodium bicarbonate (40×3 mL), and sodium chloride (40×3 mL). The combined organic phase was dried with anhydrous sodium sulfate for 1 hour, then distillated under reduced pressure and separated by column chromatography (DCM: CH.sub.3OH, 40:1 v/v) to obtain white powder solid Compound 1-6 1.3 g, yield 83%.
Synthesis of Compound 1-7
[0092] Compound 1-6 (1.5 g, 3.5 mmol) was dissolved in dichloromethane, 4 M HCl dioxane solution (9 mL, 35 mmol) was added at 0° C., and the reaction was stirred at room temperature for 12 hours. Then the solution was evaporated to dryness to obtain Intermediate 1-7 and used directly to the next step.
Synthesis of Compound 1-9
[0093] Compound 1-8 (0.76 g, 3.5 mmol) was dissolved in dichloromethane (40 mL), and the reaction solution was cooled to −20° C., then HATU (1.9 g, 4.9 mmol) was added to the reaction solution. After stirred for twenty minutes, Intermediate 1-7 obtained in the previous step was added to the reaction solution, then stirred at −20° C. for 30 minutes. DIPEA (1.7 mL, 10.5 mmol) was added dropwise to the reaction solution. After the reaction was stirred for 12 hours, it was extracted with ammonium chloride (40×3 mL), sodium bicarbonate (40×3 mL) and sodium chloride (40×3 mL). The combined organic phase was dried with anhydrous sodium sulfate for 1 hour, then distillated under reduced pressure and separated by column chromatography (DCM: CH.sub.3OH, 40:1 v/v), to obtain white powder solid Compound 1-9 1.6 g, yield 85%.
Synthesis of Compound 1-10
[0094] Intermediate 1-9 (1.86 g, 3.5 mmol) was dissolved in dichloromethane, 4 M HCl dioxane solution (9 mL, 35 mmol) was added at 0° C., and the reaction was stirred at room temperature for 12 h. Then the solution was evaporated to dryness to obtain Intermediate 1-10 and used directly in the next step.
Synthesis of Compounds 1-12
[0095] Compound 1-11 (0.61 g, 3.5 mmol) was dissolved in dichloromethane (40 mL), the reaction solution was cooled to −20° C., then HATU (1.9 g, 4.9 mmol) was added to the reaction solution. After stirred for twenty minutes, Intermediate (10) obtained in the previous step was added to the reaction solution, stirred at −20° C. for 30 minutes, and then DIPEA (1.7 mL, 10.5 mmol) was added dropwise to the reaction solution. After stirred for 12 hours, the reaction was extracted with ammonium chloride (40×3 mL), sodium bicarbonate (40×3 mL) and sodium chloride (40×3 mL). The combined organic phase was dried with anhydrous sodium sulfate for 1 hour, distillated under reduced pressure and separated by column chromatography (DCM: CH.sub.3OH, 20:1 v/v) to obtain white powder solid compound (12) 1.7 g, yield 81%.
Synthesis of Compound 1-13
[0096] Compound 1-12 (304 mg, 0.51 mmol) was dissolved in 20 ml of dichloromethane, sodium borohydride (107 mg, 2.9 mmol) was slowly added in batches, then 1 ml of methanol was added dropwise. The mixture was stirred at room temperature for about 2 hours. After the reaction was completed, about 20 ml of saturated brine was added to quench the reaction, and dichloromethane was added for extraction. The organic phase was washed with saturated brine, dried with anhydrous sodium sulfate, and separated by column chromatography (DCM: CH.sub.3OH, 20:1 v/v) to obtain white solid (13) 216 mg, yield 74%.
Synthesis of Compound 1
[0097] Compound 1-12 (165 mg, 0.29 mmol) was dissolved in 20 ml of dichloromethane, and Dess-Martin oxidant (147 mg, 0.35 mmol) was added, and stirred at room temperature. After TLC monitoring (UV) showed that the reaction was completed, the solution was extracted with sodium thiosulfate until clearation, then dried with anhydrous sodium sulfate and concentrated, column chromatography separated (DCM: CH.sub.3OH, 20:1 v/v) to obtain Compound 1 98 mg, yield 60%.
[0098] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 7.75 (d, J=9.0 Hz, 1H), 7.66-7.57 (m, 3H), 7.25 (t, J=7.8 Hz, 1H), 7.07 (s, 1H), 7.00-6.93 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.91 (s, 2H), 4.76 (dd, J=9.1, 6.5 Hz, 1H), 4.28-4.14 (m, 2H), 3.29 (m, 1H), 3.16 (m, 1H), 2.35 (m, 1H), 2.11-1.95 (m, 3H), 1.85-1.43 (m, 10H), 1.40-1.18 (m, 5H), 0.67 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 566.31[M+H].sup.+
Example 2: Synthesis of Compound 2
[0099] ##STR00095##
[0100] The Compound 2 was synthesized according to the synthesis of Compound 1 by using Compound 2-1 to replace the acid 1-11 in Example 1.
[0101] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 7.73 (s, 1H), 7.67-7.59 (m, 3H), 7.31-7.23 (m, 2H), 6.91 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.91 (s, 2H), 4.67 (dd, J=9.1, 6.5 Hz, 1H), 4.44 (dt, J=8.8, 6.4 Hz, 1H), 4.29 (dt, J=9.3, 6.9 Hz, 1H), 3.35-3.19 (m, 2H), 2.53 (m, 1H), 2.18 (dt, J=12.7, 6.3 Hz, 1H), 2.13-2.00 (m, 1H), 1.97 (dt, J=12.9, 6.4 Hz, 1H), 1.91-1.68 (m, 4H), 1.65-1.59 (m, 1H), 1.59-1.44 (m, 3H), 1.44-1.32 (m, 6H), 1.23 (m, 2H), 0.67 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 600.27[M+H].sup.+
Example 3: Synthesis of Compound 3
[0102] ##STR00096##
[0103] The Compound 3 was synthesized according to the synthesis of Compound 1 by using Compound 2-1 to replace the acid 1-11 in Example 1, and using 3-1 to replace the 1-5 in Example 1.
[0104] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 7.82 (d, J=9.2 Hz, 1H), 7.74 (s, 1H), 7.62 (dd, J=9.2, 2.2 Hz, 2H), 7.28 (d, J=8.4 Hz, 1H), 7.23 (s, 1H), 6.91 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.91 (s, 2H), 4.78 (dd, J=9.1, 6.5 Hz, 1H), 4.32 (dt, J=9.0, 6.4 Hz, 1H), 4.19 (dt, J=9.3, 6.9 Hz, 1H), 3.29 (m, 1H), 3.17 (m, 1H), 2.35 (m, 1H), 2.09 (dt, J=12.8, 6.3 Hz, 1H), 2.05-1.94 (m, 1H), 1.94-1.87 (m, 1H), 1.87-1.77 (m, 2H), 1.77-1.67 (m, 3H), 1.67-1.64 (m, 1H), 1.64-1.56 (m, 3H), 1.56-1.45 (m, 4H), 0.67 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 586.26[M+H].sup.+
Example 4: Synthesis of Compound 4
[0105] ##STR00097##
[0106] The Compound 4 was synthesized according to the synthesis of Compound 3 by using Compound 4-1 to replace the 3-1 in Example 3.
[0107] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 7.77 (d, J=9.3 Hz, 1H), 7.66-7.58 (m, 3H), 7.41 (s, 1H), 7.31-7.22 (m, 3H), 6.93 (t, J=8.1 Hz, 2H), 6.82 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.97 (dd, J=9.2, 6.6 Hz, 1H), 4.91 (s, 2H), 4.73 (dt, J=9.3, 7.7 Hz, 1H), 4.40 (dt, J=9.0, 6.4 Hz, 1H), 3.33 (m, 1H), 3.21 (m, 1H), 3.04 (dd, J=14.0, 7.8 Hz, 1H), 2.92 (dd, J=14.0, 7.8 Hz, 1H), 2.44 (m, 1H), 2.16-2.01 (m, 2H), 1.96-1.81 (m, 2H), 1.73 (m, 1H), 0.86 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 612.22[M+H].sup.+
Example 5: Synthesis of Compound 5
[0108] ##STR00098##
[0109] The Compound 5 was synthesized according to the synthesis of Compound 1 by using Compound 5-1 to replace the acid 1-11 in Example 1.
[0110] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.12 (s, 1H), 8.55 (d, J=9.2 Hz, 1H), 7.69-7.55 (m, 3H), 7.44-7.32 (m, 2H), 7.28-7.12 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 5.11 (dd, J=9.2, 6.6 Hz, 1H), 4.50-4.23 (m, 2H), 3.34 (m, 1H), 3.20 (m, 1H), 2.40 (m, 1H), 2.29-2.19 (m, J=6.7 Hz, 1H), 2.14 (dt, J=12.9, 6.4 Hz, 1H), 1.99-1.70 (m, 5H), 1.58-1.32 (m, 10H), 1.26-1.18 (m, 2H), 0.87 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 552.31[M+H].sup.+
Example 6: Synthesis of Compound 6
[0111] ##STR00099##
[0112] The Compound 6 was synthesized according to the synthesis of Compound 1 by using Compound 6-1 to replace the acid 1-11 in Example 1.
[0113] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 8.18 (d, J=9.2 Hz, 1H), 7.86 (d, J=9.3 Hz, 1H), 7.65 (d, J=9.0 Hz, 1H), 6.58-6.52 (m, 2H), 5.18 (dd, J=9.1, 6.5 Hz, 1H), 4.41 (dt, J=8.8, 6.3 Hz, 1H), 4.33 (dt, J=9.3, 6.9 Hz, 1H), 3.31 (m, 1H), 3.19 (m, 1H), 2.47 (s, 3H), 2.39 (m, 1H), 2.18-2.04 (m, 2H), 1.92 (m, 2H), 1.82 (m, 1H), 1.78-1.69 (m, 2H), 1.55-1.43 (m, 5H), 1.43-1.31 (m, 5H), 1.19 (m, 2H), 0.88 (dd, J=24.9, 6.6 Hz, 6H). ESI-MS m/z 517.29 [M+H].sup.+
Example 7: Synthesis of Compound 7
[0114] ##STR00100##
[0115] The Compound 7 was synthesized according to the synthesis of Compound 1 by using Compound 5-1 to replace the acid 1-11 in Example 1, and using Compound 7-1 to replace the 1-8 in Example 1.
[0116] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.28 (d, J=9.0 Hz, 1H), 7.68 (d, J=9.3 Hz, 1H), 7.65 (d, J=8.8 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 6.39 (d, J=6.0 Hz, 1H), 6.30 (dd, J=8.7, 7.3 Hz, 1H), 6.21 (dd, J=8.7, 7.3 Hz, 1H), 6.14 (d, J=8.6 Hz, 1H), 6.06 (dd, J=8.7, 5.7 Hz, 1H), 5.84 (dd, J=8.7, 5.3 Hz, 1H), 4.61 (dd, J=25.3, 9.0 Hz, 1H), 4.50 (dt, J=9.3, 6.9 Hz, 1H), 4.31 (dt, J=8.8, 6.4 Hz, 1H), 3.77 (dt, J=8.6, 5.2 Hz, 1H), 3.37-3.29 (m, 2H), 3.20 (m, 1H), 2.41 (m, 1H), 2.13 (dt, J=12.7, 6.3 Hz, 1H), 1.95-1.71 (m, 5H), 1.65 (t, J=25.1 Hz, 6H), 1.59-1.34 (m, 9H), 1.29 (m, 2H). ESI-MS m/z 572.32[M+H].sup.+.
Example 8: Synthesis of Compound 8
[0117] ##STR00101##
[0118] The Compound 8 was synthesized according to the synthesis of Compound 1 by using Compound 6-1 to replace the acid 1-11 in Example 1, and using compound 8-1 to replace the 1-5 in Example 1.
[0119] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.35 (s, 1H), 8.30 (d, J=6.8 Hz, 1H), 7.81-7.69 (m, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.31-7.22 (m, 4H), 7.17 (t, J=7.1 Hz, 1H), 6.94 (s, 1H), 6.52 (d, J=0.7 Hz, 1H), 4.79 (m, 1H), 4.54-4.43 (m, 1H), 4.27 (m, 1H), 3.32-3.18 (m, 3H), 3.04 (dd, J=13.8, 8.1 Hz, 1H), 2.44-2.28 (m, 2H), 2.22 (dd, J=13.4, 6.7 Hz, 1H), 1.93 (d, J=5.6 Hz, 1H), 1.81-1.72 (m, 2H), 1.31 (s, 2H), 0.93 (dd, J=10.3, 6.8 Hz, 6H). ESI-MS m/z 511.24 [M+H].sup.+.
Example 9: Synthesis of Compound 9
[0120] ##STR00102##
[0121] The Compound 9 was synthesized according to the synthesis of Compound 1 by using Compound 5-1 to replace the acid 1-11 in Example 1, and using Compound 8-1 to replace the 1-5 in Example 1.
[0122] .sup.1H NMR (600 MHz, Acetone-d6) δ 11.20 (s, 1H), 9.46 (s, 1H), 8.22 (d, J=7.4 Hz, 1H), 7.96-7.88 (m, 1H), 7.75 (dd, J=11.9, 4.0 Hz, 1H), 7.62 (m, 2H), 7.33-7.29 (m, 1H), 7.26-7.20 (m, 3H), 7.19-7.06 (m, 4H), 7.03 (dd, J=16.8, 9.5 Hz, 1H), 4.88-4.80 (m, 1H), 4.54-4.32 (m, 2H), 3.38 (dd, J=19.0, 10.1 Hz, 1H), 3.32-3.24 (m, 2H), 3.00 (m, 1H), 2.43 (m, 1H), 2.32 (m, 2H), 2.23 (m, 1H), 1.87-1.81 (m, 1H), 1.31 (d, J=1.7 Hz, 1H), 0.96 (dd, J=10.7, 6.9 Hz, 6H). ESI-MS m/z 546.26[M+H].sup.+.
Example 10: Synthesis of Compound 10
[0123] ##STR00103##
[0124] The Compound 10 was synthesized according to the synthesis of Compound 1 by using Compound 10-1 to replace the acid 1-11 in Example 1, and using Compound 8-1 to replace the 1-5 in Example 1.
[0125] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 2H), 8.32 (d, J=9.2 Hz, 2H), 7.74-7.67 (m, 4H), 7.63 (dd, J=12.5, 8.3 Hz, 4H), 7.58 (d, J=9.3 Hz, 2H), 7.39 (t, J=7.7 Hz, 2H), 7.33-7.23 (m, 7H), 7.23-7.18 (m, 1H), 7.16-7.08 (m, 4H), 6.55 (t, J=4.4 Hz, 2H), 5.01 (dd, J=9.2, 6.6 Hz, 2H), 4.75 (dt, J=9.3, 7.8 Hz, 2H), 4.38 (dt, J=8.8, 6.3 Hz, 2H), 3.38-3.28 (m, 4H), 3.26-3.10 (m, 4H), 2.42 (m, 2H), 2.28-2.16 (m, J=6.6 Hz, 2H), 2.13 (dt, J=12.8, 6.3 Hz, 2H), 1.92 (dt, J=12.8, 6.3 Hz, 2H), 1.83 (m, 2H), 1.74 (m, 2H), 0.88 (dd, J=25.1, 6.6 Hz, 12H). ESI-MS m/z 547.25[M+H]±.
Example 11: Synthesis of Compound 11
[0126] ##STR00104##
[0127] The Compound 11 was synthesized according to the synthesis of Compound 1 by using Compound 11-1 to replace the acid 1-11 in Example 1, and using compound 8-1 to replace the 1-5 in Example 1.
[0128] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.40 (s, 1H), 8.74 (d, J=8.5 Hz, 1H), 8.52 (d, J=8.5 Hz, 1H), 8.27 (d, J=8.5 Hz, 1H), 8.14 (d, J=8.5 Hz, 1H), 8.06 (d, J=8.0 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.88-7.82 (m, 1H), 7.72 (dd, J=11.2, 4.0 Hz, 1H), 7.29 (d, J=7.4 Hz, 2H), 7.19 (t, J=7.6 Hz, 2H), 7.09 (d, J=7.5 Hz, 1H), 4.85 (dd, J=11.2, 5.0 Hz, 1H), 4.62-4.55 (m, 1H), 4.40-4.30 (m, 1H), 3.33-3.20 (m, 3H), 3.06-3.00 (m, 1H), 2.50-2.41 (m, 1H), 2.38-2.27 (m, 2H), 2.06-2.00 (m, 1H), 1.80 (dt, J=12.9, 9.6 Hz, 2H), 1.30 (d, J=5.1 Hz, 1H), 0.99 (dd, J=19.0, 6.8 Hz, 6H). ESI-MS m/z 547.25[M+H].sup.+.
Example 12: Synthesis of Compound 12
[0129] ##STR00105##
synthetic route:
##STR00106## ##STR00107##
Synthesis of Compound 12-2
[0130] Compound 12-1 (1.05 g, 3.5 mmol) was dissolved in dichloromethane (40 mL), the reaction solution was cooled to −20° C., then HATU (1.9 g, 4.9 mmol) was added to the reaction solution. After stirred for twenty minutes, Compound 1-4 was added to the reaction solution, stirred at −20° C. for 30 minutes, and then DIPEA (1.7 mL, 10.5 mmol) was added dropwise to the reaction solution. After stirred for 12 hours, the reaction was extracted with ammonium chloride (40×3 mL), sodium bicarbonate (40×3 mL), and sodium chloride (40×3 mL). The combined organic phases was dried with anhydrous sodium sulfate for 1 hour, then distillated under reduced pressure and separated by column chromatography (DCM: CH.sub.3OH, 40:1 v/v) to obtain white powder solid Compound 12-2 1.3 g, yield 79%.
Synthesis of Compound 12-3
[0131] Compound 12-2 (1.64 g, 3 5 mmol) was dissolved in dichloromethane, 4 M HCl dioxane solution (9 mL, 35 mmol) was added at 0° C., and the reaction was stirred at room temperature for 12 hours. The solution was evaporated to dryness to obtain Compound 12-3 and used directly in the next step.
Synthesis of Compound 12-4
[0132] Compound 5-1 (0.56 g, 3.5 mmol) was dissolved in dichloromethane (40 mL), the reaction solution was cooled to −20° C., then HATU (1.9 g, 4.9 mmol) was added to the reaction solution. After stirred for twenty minutes, Intermediate 12-3 obtained in the previous step was added to the reaction solution, stirred at −20° C. for 30 minutes, and then DIPEA (1.7 mL, 10.5 mmol) was added dropwise to the reaction solution. After stirred for 12 hours, the reaction was extracted with ammonium chloride (40×3 mL), sodium bicarbonate (40×3 mL), and sodium chloride (40×3 mL). The combined organic phases was dried with anhydrous sodium sulfate for 1 hour, then distillated under reduced pressure and separated by column chromatography (DCM: CH.sub.3OH, 40:1 v/v) to obtain white powder solid Compound 12-4 1.4 g, with a yield of 78%.
Synthesis of Compound 12-5
[0133] Compound 12-4 (261 mg, 0.51 mmol) was dissolved in 20 ml of dichloromethane, sodium borohydride (107 mg, 2.9 mmol) was slowly added in batches, then 1 ml of methanol was added dropwise, and stirred at room temperature for about 2 hours. After the reaction was completed, about 20 ml of saturated brine was added to quench the reaction, and dichloromethane was added for extraction. The organic phase was washed with saturated brine, dried with anhydrous sodium sulfate, and separated by column chromatography (DCM: CH.sub.3OH, 20:1 v/v) to obtain white solid compound 12-5 173 mg, yield 70%
Synthesis of Compound 12
[0134] Compound 12-5 (140 mg, 0.29 mmol) was dissolved in 20 ml of dichloromethane, and Dess-Martin oxidant (147 mg, 0.35 mmol) was added and stirred at room temperature. After TLC monitoring (UV) showed that the reaction was completed, the reaction solution was extracted with sodium thiosulfate until clearation, then dried with anhydrous sodium sulfate and concentrated. Column chromatography separated (DCM: CH.sub.3OH, 20:1 v/v) to obtain Compound 12 84 mg, yield 60%.
[0135] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.11 (s, 1H), 8.63 (d, J=9.3 Hz, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.66 (d, J=7.7 Hz, 1H), 7.54 (s, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.25-7.13 (m, 3H), 7.06 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 5.01 (dt, J=9.3, 7.8 Hz, 1H), 4.40 (dt, J=8.9, 6.4 Hz, 1H), 3.31 (m, 1H), 3.19 (m, 1H), 3.07 (dd, J=14.0, 7.8 Hz, 1H), 2.96 (dd, J=13.9, 7.7 Hz, 1H), 2.41 (m, 1H), 2.16 (dt, J=12.8, 6.3 Hz, 1H), 1.92 (dt, J=12.9, 6.4 Hz, 1H), 1.84-1.70 (m, 2H). ESI-MS m/z 482.18[M+H].sup.+.
Example 13: Synthesis of Compound 13
[0136] ##STR00108##
[0137] The Compound 13 was synthesized according to the synthesis of Compound 11 by using Compound 13-1 to replace the Compound 11-1 in Example 11, and using Compound 15-1 to replace the 1-5 in Example 1.
[0138] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.37 (s, 1H), 9.21 (s, 1H), 8.47 (d, J=9.1 Hz, 1H), 7.63 (d, J=8.8 Hz, 1H), 7.58 (d, J=9.3 Hz, 1H), 7.50 (d, J=8.4 Hz, 1H), 7.38 (s, 1H), 7.29 (s, 1H), 7.28-7.20 (m, 3H), 7.14-7.09 (m, 2H), 6.80 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.98 (dd, J=9.2, 6.6 Hz, 1H), 4.74 (dt, J=9.3, 7.8 Hz, 1H), 4.39 (dt, J=8.8, 6.4 Hz, 1H), 3.83 (s, 3H), 3.39-3.29 (m, 2H), 3.27-3.10 (m, 2H), 2.44 (m, 1H), 2.26 (m, 1H), 2.11 (dt, J=12.7, 6.3 Hz, 1H), 1.94 (dt, J=12.9, 6.4 Hz, 1H), 1.83 (m, 1H), 1.75 (m, 1H), 0.88 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 576.27[M+H].sup.+.
Example 14: Synthesis of Compound 14
[0139] ##STR00109##
[0140] The Compound 14 was synthesized according to the synthesis of Compound 11 by using Compound 14-1 to replace the Compound 11-1 in Example 11.
[0141] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.04 (s, 1H), 8.44 (d, J=9.2 Hz, 1H), 7.67 (s, 1H), 7.63 (dd, J=9.1, 3.9 Hz, 2H), 7.43 (d, J=10.3 Hz, 2H), 7.28-7.20 (m, 3H), 7.14-7.08 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.93 (dd, J=9.2, 6.6 Hz, 1H), 4.71 (dt, J=9.3, 7.7 Hz, 1H), 4.42 (dt, J=9.0, 6.4 Hz, 1H), 3.38-3.29 (m, 2H), 3.23-3.11 (m, 2H), 2.40 (m, 1H), 2.18 (m, 2H), 1.97 (dt, J=12.9, 6.4 Hz, 1H), 1.85 (m, 1H), 1.74 (m, 1H), 0.90 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 614.18[M+H].sup.+
Example 15: Synthesis of Compound 15
[0142] ##STR00110##
[0143] The Compound 15 was synthesized according to the synthesis of Compound 11 by using Compound 5-1 to replace the acid 1-11 in Example 1.
[0144] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 2H), 9.12 (s, 2H), 8.54 (d, J=9.0 Hz, 2H), 7.64 (dd, J=12.7, 8.3 Hz, 4H), 7.58 (d, J=9.3 Hz, 2H), 7.42-7.36 (m, 4H), 7.22 (t, J=7.5 Hz, 2H), 7.19-7.13 (m, 2H), 6.55 (t, J=4.4 Hz, 2H), 4.92 (dd, J=9.1, 6.5 Hz, 2H), 4.68 (m, 1H), 4.58 (m, 1H), 4.46-4.33 (m, 4H), 3.33 (m, 2H), 3.21 (m, 2H), 2.41 (m, 2H), 2.19 (m, 2H), 2.12 (dd, J=12.9, 6.4 Hz, 2H), 2.01-1.55 (m, 22H), 1.47-1.38 (m, 2H), 1.32 (m, 4H), 0.87 (dd, J=25.0, 6.7 Hz, 12H). ESI-MS m/z 569.30[M+H].sup.+
Example 16: Synthesis of Compound 16
[0145] ##STR00111##
[0146] The Compound 16 was synthesized according to the synthesis of Compound 1 by using Compound 11-1 to replace the Compound 1-11 in Example 1.
[0147] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.61 (d, J=9.2 Hz, 1H), 8.30 (s, 2H), 8.15 (d, J=8.1 Hz, 1H), 7.95 (d, J=7.5 Hz, 1H), 7.84 (dd, J=8.2, 7.4 Hz, 1H), 7.72-7.49 (m, 3H), 6.55 (t, J=4.4 Hz, 1H), 4.78 (dd, J=9.0, 6.6 Hz, 1H), 4.46-4.28 (m, 2H), 3.33 (m, 1H), 3.23 (m, 1H), 2.54 (m, 1H), 2.27-2.16 (m, J=6.6 Hz, 1H), 2.11 (dt, J=12.8, 6.3 Hz, 1H), 2.02-1.90 (m, 3H), 1.89-1.80 (m, 2H), 1.74 (m, 1H), 1.62-1.47 (m, 5H), 1.45-1.24 (m, 5H), 0.88 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 564.31[M+H].sup.+.
Example 17: Synthesis of Compound 17
[0148] ##STR00112##
[0149] The Compound 17 was synthesized according to the synthesis of Compound 1 by using Compound 10-1 to replace the Compound 1-11 in Example 1.
[0150] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.32 (d, J=9.2 Hz, 1H), 7.71 (s, 1H), 7.68-7.56 (m, 4H), 7.39 (t, J=7.7 Hz, 1H), 7.29 (t, J=7.5 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.82 (dd, J=9.0, 6.6 Hz, 1H), 4.50 (dt, J=9.3, 6.9 Hz, 1H), 4.37 (dt, J=8.8, 6.3 Hz, 1H), 3.32 (m, 1H), 3.19 (m, 1H), 2.40 (m, 1H), 2.22-2.08 (m, 2H), 1.92 (dt, J=12.9, 6.4 Hz, 1H), 1.88-1.79 (m, 2H), 1.78-1.68 (m, 2H), 1.58 (m, 2H), 1.54-1.37 (m, 7H), 1.27 (m, 2H), 0.88 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 553.29[M+H].sup.+.
Example 18: Synthesis of Compound 18
[0151] ##STR00113##
[0152] The Compound 18 was synthesized according to the synthesis of Compound 1 by using Compound 18-1 to replace the Compound 1-11 in Example 1.
[0153] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.44 (d, J=9.2 Hz, 1H), 7.66-7.58 (m, 3H), 7.55 (d, J=7.9 Hz, 1H), 7.46 (d, J=7.9 Hz, 1H), 7.19 (t, J=7.8 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.85 (dd, J=9.2, 6.6 Hz, 1H), 4.41 (dt, J=9.0, 6.3 Hz, 1H), 4.34 (dt, J=9.3, 6.9 Hz, 1H), 3.33 (m, 1H), 3.23 (m, 1H), 2.51 (m, 1H), 2.26 (m, 1H), 2.12 (dt, J=12.8, 6.3 Hz, 1H), 1.94 (dt, J=12.9, 6.4 Hz, 1H), 1.86-1.43 (m, 11H), 1.39-1.17 (m, 6H), 0.90 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 631.20[M+H].sup.+.
Example 19: Synthesis of Compound 18
[0154] ##STR00114##
[0155] The Compound 19 was synthesized according to the synthesis of Compound 1 by using Compound 19-1 to replace the Compound 1-11 in Example 1.
[0156] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.36 (d, J=9.0 Hz, 1H), 8.28 (s, 1H), 8.07 (d, J=8.1 Hz, 1H), 7.81-7.74 (m, 2H), 7.62 (dd, J=9.2, 4.4 Hz, 2H), 7.52 (t, J=7.7 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.74 (dd, J=9.2, 6.6 Hz, 1H), 4.38 (dt, J=9.3, 6.9 Hz, 1H), 4.29 (dt, J=8.8, 6.4 Hz, 1H), 3.35 (m, 1H), 3.18 (m, 1H), 2.49 (s, 3H), 2.38 (m, 1H), 2.29-2.10 (m, 2H), 1.99-1.90 (m, 1H), 1.87-1.69 (m, 4H), 1.66-1.54 (m, 3H), 1.54-1.34 (m, 6H), 1.30-1.20 (m, 2H), 0.88 (dd, J=25.1, 6.6 Hz, 6H). ESI-MS m/z 578.33[M+H].sup.+.
Example 20: Synthesis of Compound 20
[0157] ##STR00115##
[0158] The Compound 20 was synthesized according to the synthesis of Compound 1 by using Compound 20-1 to replace the Compound 1-11 in Example 1.
[0159] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 8.31 (d, J=9.0 Hz, 1H), 7.82 (s, 1H), 7.76-7.71 (m, 2H), 7.60 (dd, J=11.4, 9.1 Hz, 2H), 7.42 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 5.16 (dd, J=9.0, 6.6 Hz, 1H), 4.48-4.29 (m, 2H), 3.27 (m, 2H), 2.41 (m, 1H), 2.21 (m, 1H), 2.13 (dt, J=12.8, 6.3 Hz, 1H), 2.00-1.86 (m, 2H), 1.83 (m, 1H), 1.78-1.67 (m, 3H), 1.50 (m, 2H), 1.47-1.33 (m, 6H), 1.18 (m, 2H), 0.88 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 587 0.26 [M+H].sup.+.
Example 21: Synthesis of Compound 21
[0160] ##STR00116##
[0161] The Compound 21 was synthesized according to the synthesis of Compound 12 by using Compound 21-1 to replace the Compound 12-1 in Example 12.
[0162] .sup.1H NMR (500 MHz, Chloroform) δ 9.72 (s, 4H), 8.62 (s, 4H), 8.40 (s, 4H), 7.98 (s, 4H), 7.60 (s, 4H), 7.23 (s, 4H), 7.10 (s, 3H), 6.98 (s, 3H), 6.03 (s, 4H), 5.82 (s, 4H), 5.70 (s, 4H), 5.04 (s, 2H), 3.65 (s, 2H), 3.55 (s, 2H), 2.87 (d, J=12.0 Hz, 5H), 2.61 (s, 3H), 2.23 (s, 3H), 2.02 (s, 4H), 1.93 (d, J=15.1 Hz, 6H). ESI-MS m/z 395.16[M+H].sup.+.
Example 22: Synthesis of Compound 22
[0163] ##STR00117##
[0164] The Compound 22 was synthesized according to the synthesis of Compound 12 by using Compound 3-1 to replace the Compound 12-1 in Example 12 .sup.1H NMR (500 MHz, Chloroform) δ 9.72 (s, 7H), 8.63 (s, 7H), 7.98 (s, 7H), 7.60 (s, 7H), 7.10 (s, 6H), 7.00 (d, J=18.2 Hz, 14H), 6.62 (s, 7H), 6.16 (s, 7H), 6.02 (s, 7H), 5.29 (s, 5H), 4.65 (s, 4H), 3.65 (s, 4H), 3.55 (s, 4H), 2.80 (s, 4H), 2.22 (s, 5H), 2.02 (s, 13H), 1.91 (s, 3H), 1.76 (s, 13H), 1.64 (d, J=15.3 Hz, 18H), 1.53 (s, 3H), 1.33 (s, 7H). ESI-MS m/z 439.23 [M+H]±.
Example 23: Synthesis of Compound 23
[0165] ##STR00118##
[0166] The Compound 23 was synthesized according to the synthesis of Compound 12 by using Compound 8-1 to replace the Compound 12-1 in Example 12, and using Compound 23-1 to replace the 5-1 in Example 1.
[0167] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.31 (d, J=9.0 Hz, 1H), 7.75 (s, 1H), 7.61 (m, 3H), 7.41 (d, J=8.1 Hz, 1H), 7.29-7.19 (m, 3H), 7.16-7.06 (m, 3H), 6.55 (t, J=4.4 Hz, 1H), 4.84 (dt, J=9.3, 7.8 Hz, 1H), 4.68 (dd, J=9.2, 6.6 Hz, 1H), 4.36 (dt, J=9.0, 6.4 Hz, 1H), 3.38-3.27 (m, 2H), 3.22-3.10 (m, 2H), 2.39 (m, 1H), 2.14 (dt, J=12.8, 6.3 Hz, 1H), 2.05 (m, 1H), 1.91 (dt, J=12.7, 6.3 Hz, 1H), 1.83 (m, 1H), 1.74 (m, 1H), 0.88 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 564.24[M+H].sup.+.
Example 24: Synthesis of Compound 24
[0168] ##STR00119##
[0169] The Compound 24 was synthesized according to the synthesis of Compound 1 by using Compound 24-1 to replace the Compound 1-11 in Example 1.
[0170] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 8.30 (s, 1H), 8.24 (d, J=9.0 Hz, 1H), 7.99-7.93 (m, 1H), 7.80-7.75 (m, 1H), 7.63 (d, J=9.0 Hz, 1H), 7.58 (d, J=9.3 Hz, 1H), 7.37-7.32 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 5.16 (dd, J=9.0, 6.6 Hz, 1H), 4.39 (m, 2H), 3.33 (m, 1H), 3.21 (m, 1H), 2.42 (m, 1H), 2.21-2.08 (m, 2H), 1.99-1.87 (m, 2H), 1.83 (m, 1H), 1.78-1.66 (m, 3H), 1.52 (m, 2H), 1.47-1.28 (m, 6H), 1.20 (m, 2H), 0.86 (dd, J=25.1, 6.6 Hz, 6H). ESI-MS m/z 569.27[M+H].sup.+.
Example 25: Synthesis of Compound 25
[0171] ##STR00120##
[0172] The Compound 25 was synthesized according to the synthesis of Compound 1 by using Compound 25-1 to replace the Compound 1-11 in Example 1.
Example 26: Synthesis of Compound 26
[0173] ##STR00121##
[0174] The Compound 26 was synthesized according to the synthesis of Compound 1 by using Compound 26-1 to replace the Compound 1-11 in Example 1.
[0175] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.60 (d, J=9.0 Hz, 1H), 8.30-8.19 (m, 3H), 7.90 (d, J=8.4 Hz, 1H), 7.70 (d, J=8.4 Hz, 1H), 7.63 (dd, J=9.2, 4.9 Hz, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.90 (dd, J=9.1, 6.5 Hz, 1H), 4.41 (dt, J=9.0, 6.4 Hz, 1H), 4.30 (dt, J=9.3, 6.9 Hz, 1H), 3.34 (m, 1H), 3.23 (m, 1H), 2.47 (m, 1H), 2.26-2.16 (m, J=6.6 Hz, 1H), 2.12 (dt, J=12.9, 6.4 Hz, 1H), 1.94 (dt, J=12.9, 6.4 Hz, 1H), 1.90-1.78 (m, 3H), 1.74 (m, 1H), 1.68-1.57 (m, 1H), 1.54-1.41 (m, 4H), 1.41-1.34 (m, 4H), 1.22 (m, 2H), 0.90 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 642.22[M+H].sup.+.
Example 27: Synthesis of Compound 27
[0176] ##STR00122##
[0177] The Compound 27 was synthesized according to the synthesis of Compound 1 by using Compound 24-1 to replace the Compound 1-11 in Example 1, and using Compound 8-1 to replace the 1-5 in Example 1.
[0178] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.34 (s, 1H), 8.20 (d, J=9.2 Hz, 1H), 7.98-7.93 (m, 1H), 7.80-7.75 (m, 1H), 7.61 (dd, J=19.5, 9.1 Hz, 2H), 7.38-7.32 (m, 2H), 7.29-7.20 (m, 3H), 7.15-7.08 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 5.14 (dd, J=9.1, 6.5 Hz, 1H), 4.74 (dt, J=9.3, 7.7 Hz, 1H), 4.39 (dt, J=8.8, 6.4 Hz, 1H), 3.42-3.26 (m, 2H), 3.24-3.07 (m, 2H), 2.34 (m, 1H), 2.20-2.06 (m, 2H), 1.94 (dt, J=12.7, 6.3 Hz, 1H), 1.82 (m, 1H), 1.74 (m, 1H), 0.86 (dd, J=25.1, 6.6 Hz, 6H). ESI-MS m/z 562.22[M+H].sup.+.
Example 28: Synthesis of Compound 28
[0179] ##STR00123##
[0180] The Compound 28 was synthesized according to the synthesis of Compound 9 by using Compound 28-1 to replace the Compound 1-8 in Example 9.
[0181] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.16 (s, 1H), 8.46 (d, J=9.2 Hz, 1H), 7.68-7.56 (m, 3H), 7.39 (d, J=7.9 Hz, 1H), 7.32 (s, 1H), 7.29-7.14 (m, 5H), 7.14-7.08 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.90 (dd, J=9.1, 6.5 Hz, 1H), 4.72 (dt, J=9.3, 7.8 Hz, 1H), 4.44 (dt, J=9.0, 6.3 Hz, 1H), 3.40-3.30 (m, 2H), 3.21 (m, 1H), 3.12 (dd, J=14.0, 7.8 Hz, 1H), 2.43 (m, 1H), 2.17 (dt, J=12.8, 6.3 Hz, 1H), 1.97-1.80 (m, 3H), 1.79-1.61 (m, 2H), 1.46 (m, 1H), 0.90-0.83 (m, 6H). ESI-MS m/z 559.28[M+H].sup.+.
Example 29: Synthesis of Compound 29
[0182] ##STR00124##
[0183] The Compound 29 was synthesized according to the synthesis of Compound 5 by using Compound 28-1 to replace the Compound 1-8 in Example 5.
[0184] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.16 (s, 1H), 8.46 (d, J=9.2 Hz, 1H), 7.68-7.60 (m, 3H), 7.44 (s, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.22 (t, J=7.5 Hz, 1H), 7.19-7.14 (m, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.83 (dd, J=9.0, 6.5 Hz, 1H), 4.41 (dt, J=8.9, 6.4 Hz, 1H), 4.30 (dt, J=9.3, 6.9 Hz, 1H), 3.38-3.10 (m, 2H), 2.42 (m, 1H), 2.08 (dt, J=12.9, 6.4 Hz, 1H), 2.00-1.91 (m, 2H), 1.86-1.62 (m, 5H), 1.60-1.42 (m, 7H), 1.35-1.20 (m, 5H), 0.91-0.84 (m, 6H). ESI-MS m/z 565.33[M+H].sup.+.
Example 30: Synthesis of Compound 30
[0185] ##STR00125##
[0186] The Compound 30 was synthesized according to the synthesis of Compound 1 by using Compound 5-1 to replace the acid 1-11 in Example 1, and using Compound 4-1 to replace the 1-5 in Example 1.
[0187] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.12 (s, 1H), 8.48 (d, J=9.0 Hz, 1H), 7.68-7.56 (m, 3H), 7.45 (s, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.31-7.13 (m, 4H), 7.02-6.95 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.99 (dd, J=9.2, 6.6 Hz, 1H), 4.75 (dt, J=9.2, 7.7 Hz, 1H), 4.39 (dt, J=8.8, 6.4 Hz, 1H), 3.33 (m, 1H), 3.22 (m, 1H), 3.04 (dd, J=14.1, 7.7 Hz, 1H), 2.92 (dd, J=14.0, 7.8 Hz, 1H), 2.44 (m, 1H), 2.33-2.18 (m, J=6.6 Hz, 1H), 2.12 (dt, J=12.8, 6.3 Hz, 1H), 1.94 (dt, J=12.9, 6.3 Hz, 1H), 1.83 (m, 1H), 1.74 (m, 1H), 0.88 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 563.25[M+H].sup.+.
Example 31: Synthesis of Compound 31
[0188] ##STR00126##
[0189] The Compound 31 was synthesized according to the synthesis of Compound 1 by using Compound 5-1 to replace the acid 1-11 in Example 1, and using Compound 3-1 to replace the 1-5 in Example 1.
[0190] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.12 (s, 1H), 8.55 (d, J=9.2 Hz, 1H), 7.71-7.61 (m, 3H), 7.39 (d, J=7.9 Hz, 1H), 7.32 (s, 1H), 7.24-7.14 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 5.02 (dd, J=9.1, 6.5 Hz, 1H), 4.41 (dt, J=9.0, 6.4 Hz, 1H), 4.30 (dt, J=9.3, 6.9 Hz, 1H), 3.34 (m, 1H), 3.23 (m, 1H), 2.47 (m, 1H), 2.27-2.07 (m, 2H), 1.95 (dt, J=12.9, 6.4 Hz, 1H), 1.88-1.79 (m, 3H), 1.78-1.63 (m, 4H), 1.59-1.48 (m, 5H), 0.87 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 537.29[M+H].sup.+.
Example 32: Synthesis of Compound 32
[0191] ##STR00127##
[0192] The Compound 32 was synthesized according to the synthesis of Compound 1 by using Compound 6-1 to replace the acid 1-11 in Example 1, and using Compound 4-1 to replace the 1-5 in Example 1.
[0193] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.17 (d, J=9.0 Hz, 1H), 7.98 (d, J=9.3 Hz, 1H), 7.72 (d, J=8.8 Hz, 1H), 7.28 (dd, J=8.4, 4.9 Hz, 2H), 7.03 (t, J=8.2 Hz, 2H), 6.58-6.52 (m, 2H), 4.75 (dd, J=9.0, 6.6 Hz, 1H), 4.58-4.44 (m, 2H), 3.30 (m, 1H), 3.18 (m, 1H), 3.09 (dd, J=13.9, 7.7 Hz, 1H), 2.92 (dd, J=14.1, 7.7 Hz, 1H), 2.55-2.47 (m, 4H), 2.24-2.11 (m, J=6.6 Hz, 1H), 2.01 (dt, J=12.8, 6.3 Hz, 1H), 1.87-1.76 (m, 2H), 1.73 (m, 1H), 0.88 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 529.23[M+H].sup.+.
Example 33: Synthesis of Compound 33
[0194] ##STR00128##
[0195] The Compound 33 was synthesized according to the synthesis of Compound 9 by using Compound 33-1 to replace the Compound 1-8 in Example 9.
[0196] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.12 (s, 1H), 8.49 (d, J=8.8 Hz, 1H), 7.81 (d, J=9.3 Hz, 1H), 7.69-7.60 (m, 2H), 7.41 (d, J=7.9 Hz, 1H), 7.32-7.08 (m, 8H), 6.55 (t, J=4.4 Hz, 1H), 4.79 (dt, J=9.3, 7.7 Hz, 1H), 4.48 (dt, J=9.0, 6.3 Hz, 1H), 4.42 (d, J=8.8 Hz, 1H), 3.37-3.30 (m, 2H), 3.24-3.11 (m, 2H), 2.43 (m, 1H), 2.24 (dt, J=12.7, 6.3 Hz, 1H), 1.99 (dt, J=12.9, 6.3 Hz, 1H), 1.87-1.71 (m, 2H), 0.98 (s, 9H). ESI-MS m/z 559.28[M+H].sup.+.
Example 34: Synthesis of Compound 34
[0197] ##STR00129##
[0198] The Compound 34 was synthesized according to the synthesis of Compound 9 by using Compound 33-1 to replace the Compound 1-8 in Example 5.
[0199] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.12 (s, 1H), 8.53 (d, J=8.8 Hz, 1H), 7.69-7.55 (m, 3H), 7.39 (d, J=7.9 Hz, 1H), 7.35 (s, 1H), 7.22 (t, J=7.5 Hz, 1H), 7.19-7.12 (m, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.61-4.55 (m, 2H), 4.33 (dt, J=9.3, 6.8 Hz, 1H), 3.34 (m, 1H), 3.22 (m, 1H), 2.46 (m, 1H), 2.06 (dt, J=12.8, 6.3 Hz, 1H), 1.98-1.79 (m, 3H), 1.78-1.58 (m, 5H), 1.57-1.39 (m, 3H), 1.34-1.17 (m, 5H), 0.98 (s, 9H). ESI-MS m/z 565.33[M+H].sup.+.
Example 35: Synthesis of Compound 35
[0200] ##STR00130##
[0201] The Compound 35 was synthesized according to the synthesis of Compound 1 by using Compound 5-1 to replace the acid 1-11 in Example 1, and using Compound 35-1 to replace the 1-5 in Example 1.
[0202] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.12 (s, 1H), 8.54 (d, J=9.0 Hz, 1H), 7.67-7.56 (m, 3H), 7.42-7.36 (m, 2H), 7.23 (t, J=7.6 Hz, 1H), 7.16 (dd, J=8.0, 7.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.89 (dd, J=9.0, 6.6 Hz, 1H), 4.74-4.52 (m, 2H), 4.52-4.43 (m, 1H), 4.33 (dt, J=8.8, 6.4 Hz, 1H), 3.33 (m, 1H), 3.19 (m, 1H), 2.40 (m, 1H), 2.20 (m, 1H), 2.15-2.09 (m, 1H), 2.00-1.56 (m, 12H), 0.87 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 587.29[M+H].sup.+.
Example 36: Synthesis of Compound 36
[0203] ##STR00131##
[0204] The Compound 36 was synthesized according to the synthesis of Compound 35 by using Compound 36-1 to replace the Compound 35-1 in Example 35.
[0205] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.07 (s, 1H), 8.54 (d, J=9.2 Hz, 1H), 7.73-7.62 (m, 3H), 7.39 (d, J=7.9 Hz, 1H), 7.32 (s, 1H), 7.25-7.14 (m, 2H), 6.55 (s, 1H), 4.99 (dd, J=9.2, 6.6 Hz, 1H), 4.46 (dt, J=8.8, 6.4 Hz, 1H), 4.42-4.32 (m, 1H), 3.38-3.18 (m, 2H), 2.45 (t, J=6.4 Hz, 1H), 2.25-2.15 (m, J=6.7 Hz, 1H), 2.10 (dt, J=12.9, 6.4 Hz, 1H), 2.01-1.91 (m, 2H), 1.89-1.55 (m, 10H), 0.87 (dd, J=25.0, 6.7 Hz, 6H). ESI-MS m/z 523.28[M+H].sup.+.
Example 37: Synthesis of Compound 37
[0206] ##STR00132##
[0207] The Compound 37 was synthesized according to the synthesis of Compound 12 by using Compound 8-1 to replace the Compound 12-1 in Example 12.
[0208] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.11 (s, 1H), 8.53 (d, J=9.3 Hz, 1H), 7.80 (d, J=9.0 Hz, 1H), 7.67 (d, J=7.7 Hz, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.36 (s, 1H), 7.29-7.12 (m, 7H), 6.55 (t, J=4.4 Hz, 1H), 5.04 (dt, J=9.3, 7.7 Hz, 1H), 4.36 (dt, J=8.8, 6.4 Hz, 1H), 3.25 (m, 1H), 3.15 (m, 1H), 3.11-3.01 (m, 2H), 2.41 (m, 1H), 2.18 (dt, J=12.8, 6.3 Hz, 1H), 1.87 (dt, J=12.9, 6.4 Hz, 1H), 1.82-1.68 (m, 2H). ESI-MS m/z 446.19[M+H].sup.+.
Example 38: Synthesis of Compound 38
[0209] ##STR00133##
[0210] The Compound 38 was synthesized according to the synthesis of Compound 12 by using Compound 38-1 to replace the Compound 5-1 in Example 37.
[0211] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.56 (d, J=9.3 Hz, 1H), 7.77 (d, J=9.0 Hz, 1H), 7.68 (d, J=7.8 Hz, 1H), 7.37 (d, J=8.6 Hz, 2H), 7.30 (dd, J=8.2, 7.3 Hz, 1H), 7.27-7.18 (m, 6H), 6.55 (t, J=4.4 Hz, 1H), 4.97 (dt, J=9.3, 7.8 Hz, 1H), 4.60 (dt, J=8.8, 6.4 Hz, 1H), 3.94 (s, 3H), 3.33 (m, 1H), 3.16 (m, 1H), 3.10 (dd, J=13.9, 7.7 Hz, 1H), 3.00 (dd, J=14.1, 7.7 Hz, 1H), 2.37 (m, 1H), 2.13 (dt, J=12.9, 6.4 Hz, 1H), 1.88 (dt, J=12.9, 6.4 Hz, 1H), 1.82-1.69 (m, 2H). ESI-MS m/z 460.21 [M+H].sup.+.
Example 39: Synthesis of Compound 39
[0212] ##STR00134##
[0213] The Compound 39 was synthesized according to the synthesis of Compound 12 by using Compound 39-1 to replace the Compound 5-1 in Example 37.
[0214] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.14 (s, 1H), 8.53 (d, J=9.3 Hz, 1H), 7.86 (s, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.74 (d, J=8.8 Hz, 1H), 7.52 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.27-7.18 (m, 5H), 6.55 (t, J=4.4 Hz, 1H), 4.91 (dt, J=9.3, 7.7 Hz, 1H), 4.53 (dt, J=9.0, 6.3 Hz, 1H), 3.37 (m, 1H), 3.22 (m, 1H), 3.09 (dd, J=14.0, 7.8 Hz, 1H), 3.00 (dd, J=13.9, 7.7 Hz, 1H), 2.44 (m, 1H), 2.17 (dt, J=12.9, 6.4 Hz, 1H), 1.97 (dt, J=12.9, 6.4 Hz, 1H), 1.84 (m, 1H), 1.76 (m, 1H). ESI-MS m/z 514.18[M+H].sup.+.
Example 40: Synthesis of Compound 40
[0215] ##STR00135##
[0216] The Compound 40 was synthesized according to the synthesis of Compound 12 by using Compound 13-1 to replace the Compound 5-1 in Example 37.
[0217] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 9.18 (s, 1H), 8.51 (d, J=9.3 Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.51 (d, J=8.4 Hz, 1H), 7.33 (s, 1H), 7.30 (s, 1H), 7.28-7.18 (m, 5H), 6.80 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 5.05 (dt, J=9.3, 7.8 Hz, 1H), 4.34 (dt, J=9.0, 6.3 Hz, 1H), 3.84 (s, 3H), 3.25 (m, 1H), 3.19-3.11 (m, 1H), 3.11-3.07 (m, 1H), 3.00 (dd, J=14.1, 7.7 Hz, 1H), 2.41 (m, 1H), 2.18 (dt, J=12.9, 6.4 Hz, 1H), 1.86 (dt, J=12.9, 6.4 Hz, 1H), 1.82-1.68 (m, 2H). ESI-MS m/z 476.21[M+H].sup.+.
Example 41: Synthesis of Compound 41
[0218] ##STR00136##
[0219] The Compound 41 was synthesized according to the synthesis of Compound 37 by using Compound 20-1 to replace the Compound 5-1 in Example 37.
[0220] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.15 (s, 1H), 8.53 (d, J=9.3 Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.61 (s, 1H), 7.39 (d, J=8.4 Hz, 1H), 7.32-7.07 (m, 7H), 6.55 (t, J=4.4 Hz, 1H), 5.06 (dt, J=9.3, 7.8 Hz, 1H), 4.48 (dt, J=8.8, 6.4 Hz, 1H), 3.36 (m, 1H), 3.22 (m, 1H), 3.09 (dd, J=14.1, 7.7 Hz, 1H), 3.00 (dd, J=13.9, 7.7 Hz, 1H), 2.43 (m, 1H), 2.14 (dt, J=12.9, 6.4 Hz, 1H), 1.99 (dt, J=12.9, 6.3 Hz, 1H), 1.85-1.71 (m, 2H). ESI-MS m/z 480.16[M+H].sup.+.
Example 42: Synthesis of Compound 42
[0221] ##STR00137##
[0222] The Compound 42 was synthesized according to the synthesis of Compound 37 by using Compound 42-1 to replace the Compound 5-1 in Example 37.
[0223] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.13 (s, 1H), 8.61 (d, J=9.3 Hz, 1H), 7.83 (d, J=4.9 Hz, 1H), 7.77 (d, J=9.0 Hz, 1H), 7.33 (d, J=7.8 Hz, 1H), 7.29-7.19 (m, 4H), 7.16-7.05 (m, 3H), 6.55 (t, J=4.4 Hz, 1H), 5.01 (dt, J=9.3, 7.7 Hz, 1H), 4.49 (dt, J=9.0, 6.4 Hz, 1H), 3.33 (m, 1H), 3.21-3.00 (m, 3H), 2.33 (m, 1H), 2.11 (dt, J=12.9, 6.4 Hz, 1H), 1.99 (dt, J=12.9, 6.3 Hz, 1H), 1.84-1.71 (m, 2H). ESI-MS m/z 464.19[M+H].sup.+.
Example 43: Synthesis of Compound 43
[0224] ##STR00138##
[0225] The Compound 43 was synthesized according to the synthesis of Compound 37 by using Compound 43-1 to replace the Compound 5-1 in Example 37.
[0226] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.77 (s, 1H), 9.21 (s, 1H), 8.61 (d, J=9.3 Hz, 1H), 7.77 (d, J=9.0 Hz, 1H), 7.59 (d, J=7.9 Hz, 1H), 7.39 (s, 1H), 7.32 (td, J=7.9, 4.9 Hz, 1H), 7.29-7.18 (m, 5H), 7.01 (t, J=8.0 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 5.07 (dt, J=9.3, 7.7 Hz, 1H), 4.38 (dt, J=8.8, 6.4 Hz, 1H), 3.37 (m, 1H), 3.22 (m, 1H), 3.10 (dd, J=14.0, 7.8 Hz, 1H), 3.00 (dd, J=13.9, 7.7 Hz, 1H), 2.44 (m, 1H), 2.20 (dt, J=12.8, 6.3 Hz, 1H), 1.93 (dt, J=12.9, 6.4 Hz, 1H), 1.85-1.70 (m, 2H). ESI-MS m/z 464.19[M+H].sup.+.
Example 44: Synthesis of Compound 44
[0227] ##STR00139##
[0228] The Compound 44 was synthesized according to the synthesis of Compound 37 by using Compound 44-1 to replace the Compound 5-1 in Example 37.
[0229] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.76 (s, 1H), 9.21 (s, 1H), 8.58 (d, J=9.3 Hz, 1H), 7.74 (d, J=8.8 Hz, 1H), 7.44-7.36 (m, 2H), 7.28-7.18 (m, 5H), 6.90 (t, J=8.1 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 5.06 (dt, J=9.3, 7.7 Hz, 1H), 4.41 (dt, J=9.0, 6.3 Hz, 1H), 3.37 (m, 1H), 3.22 (m, 1H), 3.10 (dd, J=13.9, 7.7 Hz, 1H), 3.00 (dd, J=14.1, 7.7 Hz, 1H), 2.44 (m, 1H), 2.19 (dt, J=12.9, 6.4 Hz, 1H), 1.92 (dt, J=12.8, 6.3 Hz, 1H), 1.85-1.70 (m, 2H). ESI-MS m/z 482.18[M+H].sup.+.
Example 45: Synthesis of Compound 45
[0230] ##STR00140##
[0231] The Compound 45 was synthesized according to the synthesis of Compound 37 by using Compound 45-1 to replace the Compound 5-1 in Example 37.
[0232] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.11 (s, 1H), 8.51 (d, J=9.3 Hz, 1H), 7.82 (d, J=9.0 Hz, 1H), 7.57 (d, J=7.9 Hz, 1H), 7.50 (dd, J=8.4, 4.9 Hz, 1H), 7.36 (s, 1H), 7.29-7.18 (m, 3H), 7.13 (dd, J=6.1, 1.1 Hz, 2H), 7.03 (t, J=8.2 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 5.06 (dt, J=9.3, 7.8 Hz, 1H), 4.48 (dt, J=9.0, 6.4 Hz, 1H), 3.36 (m, 1H), 3.22 (m, 1H), 3.09 (dd, J=14.1, 7.7 Hz, 1H), 3.00 (dd, J=13.9, 7.7 Hz, 1H), 2.44 (m, 1H), 2.14 (dt, J=12.9, 6.3 Hz, 1H), 1.99 (dt, J=12.8, 6.3 Hz, 1H), 1.85-1.70 (m, 2H). ESI-MS m/z 464.19[M+H].sup.+.
Example 46: Synthesis of Compound 46
[0233] ##STR00141##
[0234] The Compound 46 was synthesized according to the synthesis of Compound 37 by using Compound 46-1 to replace the Compound 5-1 in Example 37.
[0235] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.23 (s, 1H), 8.51 (d, J=9.3 Hz, 1H), 7.93 (dd, J=8.4, 4.9 Hz, 1H), 7.82 (d, J=9.0 Hz, 1H), 7.45 (s, 1H), 7.29-7.22 (m, 3H), 7.22-7.16 (m, 1H), 7.16-7.06 (m, 3H), 6.55 (t, J=4.4 Hz, 1H), 5.07 (dt, J=9.3, 7.8 Hz, 1H), 4.48 (dt, J=9.0, 6.4 Hz, 1H), 3.36 (m, 1H), 3.22 (m, 1H), 3.10 (dd, J=13.9, 7.7 Hz, 1H), 3.00 (dd, J=14.1, 7.7 Hz, 1H), 2.44 (m, 1H), 2.15 (dt, J=12.8, 6.3 Hz, 1H), 1.99 (dt, J=12.9, 6.3 Hz, 1H), 1.85-1.70 (m, 2H). ESI-MS m/z 464.19[M+H].sup.+.
Example 47: Synthesis of Compound 47
[0236] ##STR00142##
[0237] The Compound 47 was synthesized according to the synthesis of Compound 37 by using Compound 14-1 to replace the Compound 5-1 in Example 37, and using 1-5 to replace the compound 8-1.
[0238] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.17 (s, 1H), 8.57 (d, J=9.3 Hz, 1H), 7.81 (s, 1H), 7.74 (d, J=8.8 Hz, 1H), 7.41 (s, 1H), 7.31 (s, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.51 (dt, J=8.8, 6.4 Hz, 1H), 4.43 (dt, J=9.3, 6.9 Hz, 1H), 3.36 (m, 1H), 3.21 (m, 1H), 2.43 (m, 1H), 2.16 (dt, J=12.8, 6.3 Hz, 1H), 1.97 (dt, J=12.8, 6.3 Hz, 1H), 1.88-1.66 (m, 5H), 1.64-1.44 (m, 5H), 1.41-1.20 (m, 5H). ESI-MS m/z 520.16[M+H].sup.+.
Example 48: Synthesis of Compound 48
[0239] ##STR00143##
[0240] The Compound 48 was synthesized according to the synthesis of Compound 37 by using Compound 5-1 to replace the Compound 5-1 in Example 37, and using 1-5 to replace the Compound 8-1.
[0241] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.11 (s, 1H), 8.58 (d, J=9.2 Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.67 (d, J=7.6 Hz, 1H), 7.44 (s, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.25-7.13 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.48 (m, 2H), 3.35 (m, 1H), 3.20 (m, 1H), 2.42 (m, 1H), 2.15 (dt, J=12.9, 6.4 Hz, 1H), 2.02-1.89 (m, 2H), 1.85-1.44 (m, 9H), 1.39-1.20 (m, 5H). ESI-MS m/z 452.24[M+H].sup.+.
Example 49: Synthesis of Compound 49
[0242] ##STR00144##
[0243] The Compound 49 was synthesized according to the synthesis of Compound 47 by using Compound 38-1 to replace the Compound 14-1 in Example 47.
[0244] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.62 (d, J=9.3 Hz, 1H), 7.76 (d, J=8.8 Hz, 1H), 7.68 (d, J=7.8 Hz, 1H), 7.37 (d, J=8.3 Hz, 1H), 7.33-7.26 (m, 2H), 7.20 (t, J=7.6 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.33 (m, 2H), 3.94 (s, 3H), 3.24 (m, 1H), 3.13 (m, 1H), 2.39 (m, 1H), 2.18 (dt, J=12.9, 6.3 Hz, 1H), 1.98 (dt, J=13.8, 6.9 Hz, 1H), 1.91-1.84 (m, 2H), 1.84-1.74 (m, 2H), 1.74-1.67 (m, 1H), 1.67-1.44 (m, 7H), 1.39-1.21 (m, 5H). ESI-MS m/z 466.26[M+H].sup.+.
Example 50: Synthesis of Compound 50
[0245] ##STR00145##
[0246] The Compound 50 was synthesized according to the synthesis of Compound 47 by using Compound 50-1 to replace the Compound 14-1 in Example 47.
[0247] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.15 (s, 1H), 8.58 (d, J=9.3 Hz, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.40 (d, J=8.9 Hz, 2H), 7.27 (s, 1H), 6.80 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.55 (dt, J=9.0, 6.4 Hz, 1H), 4.42 (dt, J=9.3, 6.9 Hz, 1H), 3.84 (s, 3H), 3.35 (m, 1H), 3.20 (m, 1H), 2.46 (m, 1H), 2.13 (dt, J=12.8, 6.4 Hz, 1H), 1.98 (dt, J=12.9, 6.4 Hz, 1H), 1.86-1.65 (m, 5H), 1.64-1.53 (m, 2H), 1.53-1.43 (m, 3H), 1.40-1.20 (m, 5H). ESI-MS m/z 482.25[M+H].sup.+.
Example 51: Synthesis of Compound 51
[0248] ##STR00146##
[0249] The Compound 51 was synthesized according to the synthesis of Compound 47 by using Compound 42-1 to replace the Compound 14-1 in Example 47.
[0250] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.13 (s, 1H), 8.63 (d, J=9.3 Hz, 1H), 7.80-7.71 (m, 2H), 7.27 (d, J=7.8 Hz, 1H), 7.21 (td, J=7.8, 4.9 Hz, 1H), 7.09 (t, J=7.8 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.51 (dt, J=9.0, 6.4 Hz, 1H), 4.43 (dt, J=9.3, 6.9 Hz, 1H), 3.36 (m, 1H), 3.21 (m, 1H), 2.42 (m, 1H), 2.16 (dt, J=12.9, 6.4 Hz, 1H), 2.00 (dt, J=12.9, 6.3 Hz, 1H), 1.86-1.66 (m, 5H), 1.64-1.44 (m, 5H), 1.41-1.20 (m, 5H). ESI-MS m/z 470.23[M+H].sup.+.
Example 52: Synthesis of Compound 52
[0251] ##STR00147##
[0252] The Compound 52 was synthesized according to the synthesis of Compound 47 by using Compound 45-1 to replace the Compound 14-1 in Example 47.
[0253] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.11 (s, 1H), 8.54 (d, J=9.2 Hz, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.53 (d, J=7.9 Hz, 1H), 7.47 (dd, J=8.4, 4.9 Hz, 1H), 7.37 (s, 1H), 7.03 (t, J=8.2 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.52 (dt, J=9.0, 6.4 Hz, 1H), 4.45 (dt, J=9.3, 6.9 Hz, 1H), 3.35 (m, 1H), 3.20 (m, 1H), 2.43 (m, 1H), 2.12 (dt, J=12.8, 6.3 Hz, 1H), 1.99 (dt, J=12.9, 6.3 Hz, 1H), 1.92-1.68 (m, 4H), 1.68-1.44 (m, 7H), 1.40-1.24 (m, 5H). ESI-MS m/z 470.23[M+H].sup.+.
Example 53: Synthesis of Compound 53
[0254] ##STR00148##
[0255] The Compound 53 was synthesized according to the synthesis of Compound 47 by using Compound 46-1 to replace the Compound 14-1 in Example 47.
[0256] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.23 (s, 1H), 8.54 (d, J=9.2 Hz, 1H), 7.90 (dd, J=8.4, 4.9 Hz, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.49 (s, 1H), 7.18-7.07 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.51 (dt, J=8.8, 6.3 Hz, 1H), 4.43 (dt, J=9.3, 6.9 Hz, 1H), 3.36 (m, 1H), 3.21 (m, 1H), 2.42 (m, 1H), 2.16 (dt, J=12.9, 6.4 Hz, 1H), 1.99 (dt, J=12.8, 6.3 Hz, 1H), 1.86-1.66 (m, 5H), 1.59 (m, 2H), 1.55-1.50 (m, 2H), 1.50-1.44 (m, 2H), 1.41-1.20 (m, 5H). ESI-MS m/z 470.23[M+H].sup.+.
Example 54: Synthesis of Compound 54
[0257] ##STR00149##
[0258] The Compound 54 was synthesized according to the synthesis of Compound 47 by using Compound 43-1 to replace the Compound 14-1 in Example 47.
[0259] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.78 (s, 1H), 9.21 (s, 1H), 8.63 (d, J=9.3 Hz, 1H), 7.74 (d, J=8.8 Hz, 1H), 7.57 (d, J=7.9 Hz, 1H), 7.47 (s, 1H), 7.30 (td, J=7.9, 5.0 Hz, 1H), 7.00 (t, J=7.9 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.47-4.35 (m, 2H), 3.37 (m, 1H), 3.22 (m, 1H), 2.43 (m, 4.5 Hz, 1H), 2.21 (dt, J=12.8, 6.3 Hz, 1H), 1.94 (dt, J=12.9, 6.3 Hz, 1H), 1.88-1.66 (m, 5H), 1.64-1.44 (m, 5H), 1.41-1.21 (m, 5H). ESI-MS m/z 470.23[M+H].sup.+.
Example 55: Synthesis of Compound 55
[0260] ##STR00150##
[0261] The Compound 55 was synthesized according to the synthesis of Compound 47 by using Compound 23-1 to replace the Compound 14-1 in Example 47.
[0262] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.50 (d, J=9.3 Hz, 1H), 7.83-7.76 (m, 2H), 7.64 (dd, J=9.8, 7.7 Hz, 2H), 7.39 (t, J=7.5 Hz, 1H), 7.32 (t, J=7.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.38 (dt, J=8.8, 6.4 Hz, 1H), 4.25 (dt, J=9.3, 6.9 Hz, 1H), 3.32 (m, 1H), 3.20 (m, 1H), 2.42 (m, 1H), 2.16 (dt, J=12.9, 6.4 Hz, 1H), 1.98-1.65 (m, 6H), 1.59 (m, 2H), 1.57-1.44 (m, 3H), 1.41-1.18 (m, 5H). ESI-MS m/z 453.23[M+H].sup.+.
Example 56: Synthesis of Compound 56
[0263] ##STR00151##
[0264] The Compound 56 was synthesized according to the synthesis of Compound 47 by using Compound 24-1 to replace the Compound 14-1 in Example 47.
[0265] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 8.46 (d, J=9.3 Hz, 1H), 8.30 (s, 1H), 7.96 (d, J=7.8 Hz, 1H), 7.83-7.77 (m, 2H), 7.42 (t, J=7.6 Hz, 1H), 7.35 (t, J=7.6 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.48 (dt, J=8.8, 6.4 Hz, 1H), 4.29 (dt, J=9.3, 6.9 Hz, 1H), 3.32-3.16 (m, 2H), 2.36 (m, 1H), 2.17 (dt, J=12.9, 6.4 Hz, 1H), 1.96 (m, 2H), 1.86-1.44 (m, 10H), 1.41-1.17 (m, 6H). ESI-MS m/z 469.20[M+H].sup.+.
Example 57: Synthesis of Compound 57
[0266] ##STR00152##
[0267] The Compound 49 was synthesized according to the synthesis of Compound 47 by using Compound 38-1 to replace the Compound 14-1 in Example 47.
[0268] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 8.63 (d, J=9.3 Hz, 1H), 7.83-7.73 (m, 2H), 7.65-7.57 (m, 1H), 7.29-7.21 (m, 2H), 6.55 (t, J=4.4 Hz, 1H), 4.56-4.47 (m, 2H), 3.36 (m, 1H), 3.21 (m, 1H), 2.42 (m, 1H), 2.15 (dt, J=12.8, 6.3 Hz, 1H), 1.98 (dt, J=12.8, 6.3 Hz, 1H), 1.85-1.66 (m, 5H), 1.63-1.45 (m, 6H), 1.42-1.21 (m, 6H). ESI-MS m/z 453 0.24 [M+H]±.
Example 58: Synthesis of Compound 58
[0269] ##STR00153##
[0270] The Compound 58 was synthesized according to the synthesis of Compound 47 by using Compound 11-1 to replace the Compound 14-1 in Example 47.
[0271] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.57 (d, J=9.3 Hz, 1H), 8.35-8.26 (m, 2H), 8.15 (d, J=8.1 Hz, 1H), 7.93 (d, J=7.5 Hz, 1H), 7.85 (dd, J=8.2, 7.4 Hz, 1H), 7.71 (d, J=8.8 Hz, 1H), 7.59 (t, J=7.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.45-4.35 (m, 2H), 3.27 (m, 1H), 3.12 (m, 1H), 2.40 (m, 1H), 2.10 (dt, J=12.8, 6.3 Hz, 1H), 2.00-1.89 (m, 2H), 1.84-1.65 (m, 3H), 1.65-1.43 (m, 6H), 1.39-1.19 (m, 5H). ESI-MS m/z 464.24[M+H]±.
Example 59: Synthesis of Compound 59
[0272] ##STR00154##
[0273] The Compound 59 was synthesized according to the synthesis of Compound 47 by using Compound 59-1 to replace the Compound 14-1 in Example 47.
[0274] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.36 (s, 1H), 9.21 (s, 1H), 8.64 (d, J=9.3 Hz, 1H), 7.99 (dd, J=7.8, 3.7 Hz, 2H), 7.86 (t, J=7.6 Hz, 1H), 7.75 (d, J=9.0 Hz, 1H), 7.61-7.54 (m, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.43 (dt, J=9.3, 6.9 Hz, 1H), 4.37 (dt, J=8.8, 6.4 Hz, 1H), 3.23 (m, 1H), 3.02 (m, 1H), 2.36 (m, 1H), 2.09 (dt, J=12.7, 6.3 Hz, 1H), 1.99-1.89 (m, 2H), 1.83-1.44 (m, 10H), 1.40-1.19 (m, 6H). ESI-MS m/z 465.24[M+H].sup.+.
Example 60: Synthesis of Compound 60
[0275] ##STR00155##
[0276] The Compound 60 was synthesized according to the synthesis of Compound 9 by using Compound 60-1 to replace the Compound 1-8 in Example 5.
[0277] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.12 (s, 1H), 8.79 (d, J=9.1 Hz, 1H), 7.71-7.60 (m, 3H), 7.42-7.35 (m, 2H), 7.23 (t, J=7.6 Hz, 1H), 7.20-7.13 (m, 1H), 6.55 (t, J=4.4 Hz, 1H), 4.50 (dt, J=8.8, 6.4 Hz, 1H), 4.38 (dt, J=9.3, 6.8 Hz, 1H), 4.18 (dd, J=9.1, 5.2 Hz, 1H), 3.30 (m, 1H), 3.15 (m, 1H), 2.39 (m, 1H), 2.11 (dt, J=12.8, 6.3 Hz, 1H), 1.97 (dt, J=12.9, 6.3 Hz, 1H), 1.92-1.65 (m, 5H), 1.62-1.52 (m, 1H), 1.52-1.31 (m, 9H), 1.23-1.08 (m, 4H), 0.89-0.80 (m, 2H). ESI-MS m/z 549.29[M+H].sup.+.
Example 61: Synthesis of Compound 61
[0278] ##STR00156##
[0279] The Compound 61 was synthesized according to the synthesis of Compound 47 by using Compound 61-1 to replace the Compound 14-1 in Example 47.
[0280] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.21 (s, 1H), 8.48 (d, J=9.3 Hz, 1H), 7.66 (d, J=8.8 Hz, 1H), 7.54 (d, J=8.4 Hz, 1H), 7.42 (s, 1H), 6.88 (d, J=8.4 Hz, 1H), 6.55 (t, J=4.4 Hz, 1H), 6.05 (d, J=6.8 Hz, 2H), 4.35 (m, 2H), 3.32 (m, 1H), 3.19 (m, 1H), 2.40 (m, 1H), 2.19 (dt, J=12.8, 6.3 Hz, 1H), 1.93-1.67 (m, 6H), 1.59 (m, 2H), 1.57-1.44 (m, 3H), 1.41-1.20 (m, 6H). ESI-MS m/z 457.22[M+H].sup.+.
Example 62: Synthesis of Compound 62
[0281] ##STR00157##
[0282] The Compound 62 was synthesized according to the synthesis of Compound 47 by using Compound 6-1 to replace the Compound 14-1 in Example 47.
[0283] .sup.1H NMR (600 MHz, Acetone-d6) δ 11.14 (d, J=8.4 Hz, 1H), 9.41 (s, 1H), 8.28-8.23 (m, 1H), 7.78 (dd, J=12.5, 8.8 Hz, 1H), 7.58 (d, J=1.8 Hz, 1H), 7.47 (d, J=8.8 Hz, 1H), 7.37-7.33 (m, 2H), 7.23 (d, J=7.7 Hz, 2H), 7.16 (dd, J=6.3, 1.9 Hz, 4H), 5.44 (dd, J=70.6, 8.0 Hz, 1H), 5.06 (dd, J=7.1, 4.5 Hz, 1H), 4.51 (m, 1H), 4.13-3.99 (m, 1H), 3.32 (d, J=2.0 Hz, 1H), 3.20 (m, 2H), 2.44-2.34 (m, 1H), 2.33-2.25 (m, 1H), 1.79-1.55 (m, 2H). ESI-MS m/z 413.17[M+H].sup.+.
Example 63: Synthesis of Compound 63
[0284] ##STR00158##
[0285] The Compound 63 was synthesized according to the synthesis of Compound 47 by using Compound 63-1 to replace the Compound 14-1 in Example 47.
[0286] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.42 (s, 1H), 8.87 (d, J=12.0 Hz, 1H), 8.75 (d, J=6.0 Hz, 1H), 8.63 (d, J=2.7 Hz, 1H), 8.24 (d, J=8.1 Hz, 1H), 7.98-7.94 (m, 1H), 7.37 (d, J=7.3 Hz, 2H), 7.30 (dd, J=10.4, 4.7 Hz, 2H), 7.22 (m, 1H), 5.01 (m, 1H), 4.32-4.27 (m, 1H), 3.38-3.34 (m, 1H), 3.31-3.27 (m, 1H), 3.19 (dd, J=13.9, 8.9 Hz, 1H), 2.51-2.40 (m, 1H), 2.39-2.33 (m, 1H), 1.97-1.88 (m, 1H), 1.82 (m, 1H), 1.39 (s, 1H), 1.31 (d, J=2.5 Hz, 2H). ESI-MS m/z 427.16[M+H].sup.+.
Example 64: Synthesis of Compound 64
[0287] ##STR00159##
[0288] The Compound 64 was synthesized according to the synthesis of Compound 12 by using Compound 64-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0289] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.43 (s, 1H), 8.59 (d, J=6.8 Hz, 1H), 8.18 (dd, J=15.6, 1.8 Hz, 1H), 8.08-7.99 (m, 1H), 7.92-7.82 (m, 2H), 7.53 (t, J=8.1 Hz, 1H), 7.38 (d, J=7.3 Hz, 2H), 7.28 (t, J=7.5 Hz, 2H), 7.19 (t, J=7.3 Hz, 1H), 7.13 (s, 1H), 6.97-6.90 (m, 1H), 5.07 (m, 1H), 4.37 (m, 1H), 3.37-3.33 (m, 1H), 3.29-3.20 (m, 3H), 2.52-2.24 (m, 2H), 2.05-1.98 (m, 1H), 1.87-1.62 (m, 2H). ESI-MS m/z 448.17[M+H].sup.+.
Example 65: Synthesis of Compound 65
[0290] ##STR00160##
[0291] The Compound 65 was synthesized according to the synthesis of Compound 12 by using Compound 65-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0292] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.22 (s, 1H), 8.36 (d, J=6.3 Hz, 1H), 7.32 (d, J=2.0 Hz, 1H), 7.23 (s, 5H), 7.02 (d, J=8.1 Hz, 1H), 6.83 (d, J=8.4 Hz, 1H), 6.55 (s, 1H), 5.07 (m, 1H), 4.27-4.21 (m, 6H), 3.31-3.24 (m, 2H), 3.20 (dd, J=14.4, 6.7 Hz, 2H), 2.35-2.28 (m, 2H), 1.90 (dd, J=10.2, 6.4 Hz, 1H), 1.80 (m, 2H). ESI-MS m/z 466.18[M+H].sup.+.
Example 66: Synthesis of Compound 66
[0293] ##STR00161##
[0294] The Compound 66 was synthesized according to the synthesis of Compound 12 by using Compound 66-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0295] .sup.1H NMR (600 MHz, Acetone-d6) δ 11.14 (d, J=8.8 Hz, 1H), 9.41 (s, 1H), 8.28-8.23 (m, 1H), 7.78 (m, 1H), 7.58 (d, J=1.8 Hz, 1H), 7.47 (d, J=8.8 Hz, 1H), 7.37-7.33 (m, 2H), 7.23 (d, J=7.7 Hz, 2H), 7.16 (dd, J=6.3, 1.9 Hz, 4H), 5.44 m, 1H), 5.06 (dd, J=7.1, 4.5 Hz, 1H), 4.51 (m, 1H), 4.13-3.99 (m, 1H), 3.32 (d, J=2.0 Hz, 1H), 3.20 (m, 2H), 2.44-2.34 (m, 1H), 2.33-2.25 (m, 1H), 1.79-1.55 (m, 2H). ESI-MS m/z 466.18[M+H].sup.+.
Example 67: Synthesis of Compound 67
[0296] ##STR00162##
[0297] The Compound 67 was synthesized according to the synthesis of Compound 12 by using Compound 67-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0298] .sup.1H NMR (600 MHz, Acetone-d6) δ 10.53 (s, 1H), 9.39 (s, 1H), 8.19-8.16 (m, 1H), 7.73 (d, J=8.0 Hz, 1H), 7.66 (dd, J=8.5, 1.7 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 7.40-7.38 (m, 1H), 7.37-7.33 (m, 2H), 7.26 (m, 2H), 7.19-7.15 (m, 1H), 6.96 (s, 1H), 6.54 (m, 1H), 5.01 (dt, J=8.2, 4.0 Hz, 1H), 4.31-4.26 (m, 1H), 3.31 (dd, J=8.2, 5.6 Hz, 1H), 3.26-3.18 (m, 3H), 2.45-2.36 (m, 1H), 2.32-2.21 (m, 1H), 1.98-1.95 (m, 1H), 1.81-1.68 (m, 2H). ESI-MS m/z 447.19[M+H].sup.+.
Example 68: Synthesis of Compound 68
[0299] ##STR00163##
[0300] The Compound 68 was synthesized according to the synthesis of Compound 12 by using Compound 68-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0301] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.40 (s, 1H), 8.79 (s, 1H), 8.53 (d, J=6.2 Hz, 1H), 8.27 (d, J=4.9 Hz, 1H), 8.04 (d, J=7.3 Hz, 1H), 7.58-7.52 (m, 1H), 7.47-7.40 (m, 1H), 7.32 (t, J=6.4 Hz, 2H), 7.28-7.22 (m, 2H), 7.18 (t, J=7.2 Hz, 1H), 6.91 (s, 1H), 5.03-4.93 (m, 1H), 4.44 (m, 1H), 3.32 (t, J=6.9 Hz, 1H), 3.27-3.23 (m, 1H), 2.45-2.26 (m, 2H), 1.99-1.94 (m, 1H), 1.80-1.75 (m, 1H), 1.37 (s, 1H), 1.29 (d, J=3.3 Hz, 2H). ESI-MS m/z 526.11 [M+H].sup.+.
Example 69: Synthesis of Compound 69
[0302] ##STR00164##
[0303] The Compound 69 was synthesized according to the synthesis of Compound 12 by using Compound 24-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0304] .sup.1H NMR (600 MHz, Acetone-d6) δ 11.14 (d, J=8.4 Hz, 1H), 9.41 (s, 1H), 8.28-8.23 (m, 1H), 7.78 (dd, J=12.5, 8.8 Hz, 1H), 7.58 (d, J=1.8 Hz, 1H), 7.47 (d, J=8.8 Hz, 1H), 7.37-7.33 (m, 2H), 7.23 (d, J=7.7 Hz, 2H), 7.16 (dd, J=6.3, 1.9 Hz, 4H), 5.44 (m, 1H), 5.06 (dd, J=7.1, 4.5 Hz, 1H), 4.51 (m, 1H), 4.13-3.99 (m, 1H), 3.32 (d, J=2.0 Hz, 1H), 3.20 (m, 2H), 2.44-2.34 (m, 1H), 2.33-2.25 (m, 1H), 1.79-1.55 (m, 2H). ESI-MS m/z 464.15[M+H].sup.+.
Example 70: Synthesis of Compound 70
[0305] ##STR00165##
[0306] The Compound 70 was synthesized according to the synthesis of Compound 12 by using Compound 10-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0307] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.43 (s, 1H), 8.64 (d, J=6.8 Hz, 1H), 8.11 (d, J=8.0 Hz, 1H), 7.71 (d, J=7.5 Hz, 1H), 7.56-7.50 (m, 1H), 7.49-7.41 (m, 2H), 7.37 (dd, J=9.2, 2.3 Hz, 2H), 7.32-7.22 (m, 3H), 7.19 (d, J=7.5 Hz, 1H), 7.12 (s, 1H), 5.10-5.01 (m, 1H), 4.58-4.32 (m, 1H), 3.38 (m, 1H), 3.25 (dd, J=9.0, 2.5 Hz, 3H), 2.53-2.27 (m, 2H), 2.04 (m, 1H), 1.86-1.62 (m, 2H). ESI-MS m/z 448.17[M+H].sup.+.
Example 71: Synthesis of Compound 71
[0308] ##STR00166##
[0309] The Compound 71 was synthesized according to the synthesis of Compound 12 by using Compound 59-1 to replace the Compound 5-1 in Example 12, and using Compound 8-1 to replace the Compound 12-1 in Example 1.
[0310] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.47 (s, 1H), 8.69 (d, J=8.0 Hz, 1H), 8.15 (m, 2H), 7.95 (m, 2H), 7.80 (d, J=8.0 Hz, 1H), 7.34 (d, J=7.3 Hz, 2H), 7.25 (t, J=7.3 Hz, 2H), 7.18 (t, J=7.3 Hz, 1H), 6.83 (s, 1H), 5.03-4.95 (m, 1H), 3.99 (dd, J=7.3, 4.3 Hz, 1H), 3.50 (m, 2H), 3.31-3.20 (m, 3H), 2.39-2.28 (m, 2H), 1.95-1.87 (m, 1H), 1.78-1.68 (m, 1H), 1.62-1.51 (m, 1H). ESI-MS m/z 460.18[M+H].sup.+.
Example 72: Synthesis of Compound 72
[0311] ##STR00167##
[0312] The Compound 72 was synthesized according to the synthesis of Compound 1 by using Compound 72-1 to replace the acid 1-11 in Example 1, and using
[0313] Compound 8-1 to replace the Compound 1-5 in Example 1.
[0314] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 8.63 (d, J=8.5 Hz, 1H), 8.33 (d, J=6.2 Hz, 1H), 8.06 (d, J=7.5 Hz, 1H), 8.02 (d, J=8.5 Hz, 1H), 7.80 (m, 1H), 7.75 (dd, J=11.0, 4.0 Hz, 1H), 7.39-7.27 (m, 5H), 7.25 (d, J=7.1 Hz, 1H), 6.56 (s, 1H), 5.12 (q, J=7.1 Hz, 1H), 4.32 (m, 1H), 3.31 (dd, J=12.8, 6.2 Hz, 3H), 3.02 (s, 3H), 2.40-2.30 (m, 2H), 1.99-1.93 (m, 1H), 1.88-1.75 (m, 2H). ESI-MS m/z 474.21[M+H].sup.+.
Example 73: Synthesis of Compound 73
[0315] ##STR00168##
[0316] The Compound 73 was synthesized according to the synthesis of Compound 1 by using Compound 11-1 to replace the acid 1-11 in Example 1, and using Compound 8-1 to replace the Compound 1-5 in Example 1.
[0317] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.22 (s, 1H), 8.84 (d, J=8.5 Hz, 1H), 8.28 (d, J=8.5 Hz, 1H), 8.22 (dd, J=10.1, 6.0 Hz, 1H), 8.11 (m, 2H), 7.86 (d, J=8.1 Hz, 1H), 7.77 (dd, J=11.2, 4.1 Hz, 1H), 7.62 (m, 1H), 7.36-7.27 (m, 4H), 7.24 (t, J=7.2 Hz, 1H), 5.08 (m, 1H), 4.33-4.28 (m, 1H), 3.35-3.29 (m, 2H), 3.25 (t, J=6.8 Hz, 2H), 2.38-2.30 (m, 2H), 1.93-1.84 (m, 2H), 1.79-1.71 (m, 2H). ESI-MS m/z 459.19[M+H].sup.+.
Example 74: Synthesis of Compound 74
[0318] ##STR00169##
[0319] The Compound 74 was synthesized according to the synthesis of Compound 1 by using Compound 74-1 to replace the acid 1-11 in Example 1, and using
[0320] Compound 8-1 to replace the Compound 1-5 in Example 1.
[0321] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.46 (s, 1H), 8.91 (dd, J=7.9, 4.6 Hz, 1H), 8.71 (d, J=6.6 Hz, 1H), 8.44-8.39 (m, 1H), 8.24 (m, 1H), 8.17 (t, J=7.9 Hz, 1H), 7.98 (dd, J=14.5, 7.3 Hz, 1H), 7.90 (m, 1H), 7.42-7.38 (m, 2H), 7.30-7.25 (m, 2H), 7.20 (dd, J=10.4, 4.4 Hz, 1H), 7.02 (m, 1H), 5.16-5.07 (m, 1H), 4.62-4.32 (m, 1H), 3.42-3.37 (m, 1H), 3.30 (s, 2H), 2.53-2.30 (m, 2H), 1.99 (m, 1H), 1.88-1.62 (m, 2H). ESI-MS m/z 527.18[M+H].sup.+.
Example 75: Synthesis of Compound 75
[0322] ##STR00170##
[0323] The Compound 75 was synthesized according to the synthesis of Compound 1 by using Compound 24-1 to replace the acid 1-11 in Example 1, and using Compound 8-1 to replace the Compound 1-5 in Example 1.
[0324] .sup.1H NMR (600 MHz, Acetone-d6) δ 11.14 (d, J=8.4 Hz, 1H), 9.41 (s, 1H), 8.28-8.23 (m, 1H), 7.78 (dd, J=12.5, 8.8 Hz, 1H), 7.58 (d, J=1.8 Hz, 1H), 7.47 (d, J=8.8 Hz, 1H), 7.37-7.33 (m, 2H), 7.23 (d, J=7.7 Hz, 2H), 7.16 (dd, J=6.3, 1.9 Hz, 4H), 5.44 (m, 1H), 5.06 (dd, J=7.1, 4.5 Hz, 1H), 4.51 (m, 1H), 4.13-3.99 (m, 1H), 3.32 (d, J=2.0 Hz, 1H), 3.20 (m, 2H), 2.44-2.34 (m, 1H), 2.33-2.25 (m, 1H), 1.79-1.55 (m, 2H). ESI-MS m/z 537.10[M+H].sup.+.
Example 76: Synthesis of Compound 76
[0325] ##STR00171##
[0326] The Compound 76 was synthesized according to the synthesis of Compound 1 by using Compound 2-1 to replace the acid 1-11 in Example 1, and using Compound 8-1 to replace the Compound 1-5 in Example 1.
[0327] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.24 (s, 1H), 8.55 (d, J=6.0 Hz, 1H), 7.29-7.26 (m, 2H), 7.22 (m, 5H), 7.10 (m, 1H), 6.99 (d, J=2.6 Hz, 1H), 6.95 (s, 1H), 6.72 (d, J=8.6 Hz, 1H), 6.58 (s, 1H), 5.01 (dd, J=14.9, 6.9 Hz, 1H), 4.91-4.85 (m, 2H), 4.25-4.20 (m, 1H), 3.32-3.26 (m, 2H), 3.18 (m, 2H), 2.34-2.29 (m, 2H), 1.94-1.87 (m, 1H), 1.82-1.77 (m, 1H). ESI-MS m/z 496.15[M+H].sup.+.
Example 77: Synthesis of Compound 77
[0328] ##STR00172##
[0329] The Compound 77 was synthesized according to the synthesis of Compound 1 by using Compound 65-1 to replace the acid 1-11 in Example 1, and using
[0330] Compound 8-1 to replace the Compound 1-5 in Example 1.
[0331] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.72 (s, 1H), 8.05 (s, 1H), 7.65-7.46 (m, 3H), 7.21-7.09 (m, 3H), 5.99 (s, 1H), 5.78 (s, 1H), 5.08 (s, 1H), 4.91 (m, 2H), 4.29 (m, 4H), 3.64 (s, 1H), 3.54 (s, 1H), 3.29 (s, 1H), 3.04 (s, 1H), 2.81 (s, 1H), 2.73 (s, 1H), 2.22 (s, 1H), 2.02 (s, 1H), 1.91 (s, 1H), 0.96 (s, 6H). ESI-MS m/z 565.26[M+H].sup.+.
Example 78: Synthesis of Compound 78
[0332] ##STR00173##
[0333] The Compound 78 was synthesized according to the synthesis of Compound 1 by using Compound 13-1 to replace the acid 1-11 in Example 1.
[0334] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.69 (s, 1H), 8.57 (s, 1H), 8.39 (s, 1H), 7.56 (s, 1H), 7.42 (s, 1H), 7.14 (s, 1H), 6.89 (s, 1H), 6.77 (s, 1H), 5.95 (s, 1H), 5.43 (s, 1H), 5.16 (s, 1H), 4.92 (s, 1H), 4.49 (s, 1H), 3.80 (s, 3H), 3.64 (s, 1H), 3.53 (s, 1H), 2.85 (s, 1H), 2.72 (s, 1H), 2.22 (s, 1H), 2.02 (s, 1H), 1.91 (s, 1H), 1.75 (s, 1H), 1.60 (s, 1H), 1.54-1.36 (m, 7H), 1.29 (s, 1H), 0.96 (s, 6H). ESI-MS m/z 582.32[M+H].sup.+.
Example 79: Synthesis of Compound 79
[0335] ##STR00174##
[0336] The Compound 79 was synthesized according to the synthesis of Compound 1 by using Compound 14-1 to replace the acid 1-11 in Example 1.
[0337] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.72 (s, 1H), 8.59 (s, 1H), 7.41 (s, 1H), 7.41-7.28 (m, 3H), 7.10 (s, 1H), 6.67 (s, 1H), 6.04 (s, 1H), 4.99 (s, 1H), 4.78 (s, 1H), 4.50 (s, 1H), 3.65 (s, 1H), 3.53 (s, 1H), 2.73 (s, 1H), 2.56 (s, 1H), 2.21 (s, 1H), 2.02 (s, 1H), 1.91 (s, 1H), 1.76 (s, 2H), 1.60 (s, 1H), 1.51 (m, 3H), 1.48-1.41 (m, 4H), 1.38 (s, 1H), 0.96 (s, 6H). ESI-MS m/z 620.23 [M+H].sup.+.
Example 80: Synthesis of Compound 80
[0338] ##STR00175##
[0339] The Compound 80 was synthesized according to the synthesis of Compound 1 by using Compound 80-1 to replace the acid 1-11 in Example 1.
[0340] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.72 (s, 1H), 7.91 (s, 2H), 7.59 (s, 2H), 7.51 (s, 1H), 7.32 (m, 1H), 6.99 (s, 1H), 6.08 (s, 1H), 6.01 (s, 1H), 5.53 (s, 1H), 4.91 (m, 2H), 4.59 (s, 1H), 3.65 (s, 1H), 3.54 (s, 1H), 2.73 (m, 2H), 2.22 (s, 1H), 2.02 (s, 1H), 1.92 (s, 1H), 1.76 (s, 1H), 1.58 (s, 1H), 1.55-1.41 (m, 5H), 1.28 (s, 1H), 0.96 (s, 6H). ESI-MS m/z 539.31[M+H].sup.+.
Example 81: Synthesis of Compound 81
[0341] ##STR00176##
[0342] The Compound 81 was synthesized according to the synthesis of Compound 12 by using Compound 81-1 to replace the Compound 12-1 in Example 12.
[0343] .sup.1H NMR (600 MHz, Acetone-d6) δ 9.30 (s, 1H), 9.11 (s, 1H), 8.57 (d, J=9.3 Hz, 1H), 7.80 (d, J=9.0 Hz, 1H), 7.74 (s, 1H), 7.66 (d, J=7.6 Hz, 1H), 7.39 (d, J=7.8 Hz, 1H), 7.30 (td, J=7.9, 4.9 Hz, 1H), 7.25-7.13 (m, 2H), 6.98 (td, J=8.1, 2.9 Hz, 3H), 6.55 (t, J=4.4 Hz, 1H), 5.07 (dt, J=9.3, 7.7 Hz, 1H), 4.46 (dt, J=8.8, 6.3 Hz, 1H), 3.33 (m, 1H), 3.16 (m, 1H), 3.03 (dd, J=14.0, 7.8 Hz, 1H), 2.96 (dd, J=13.9, 7.7 Hz, 1H), 2.28 (m, 1H), 2.11 (dt, J=12.9, 6.4 Hz, 1H), 1.95 (dt, J=12.9, 6.3 Hz, 1H), 1.84-1.69 (m, 2H). ESI-MS m/z 464.19 [M+H].sup.+.
Example 82: Synthesis of Compound 82
[0344] ##STR00177##
[0345] The Compound 82 was synthesized according to the synthesis of Compound 12 by using Compound 4-1 to replace the Compound 12-1 in Example 12.
[0346] .sup.1H NMR (500 MHz, Chloroform) δ 11.66 (s, 1H), 9.72 (s, 1H), 7.98 (dt, J=14.9, 3.1 Hz, 1H), 7.60 (dd, J=14.9, 3.2 Hz, 1H), 7.31 (d, J=3.0 Hz, 1H), 7.29-6.93 (m, 7H), 6.44 (s, 1H), 5.20 (dd, J=25.8, 13.3 Hz, 1H), 5.04 (t, J=10.4 Hz, 1H), 3.76-3.48 (m, 2H), 3.29 (dd, J=24.7, 10.3 Hz, 1H), 3.04 (dd, J=24.8, 10.4 Hz, 1H), 2.59 (ddd, J=23.1, 15.0, 9.2 Hz, 1H), 2.32-2.14 (m, 1H), 2.07-1.88 (in, 3H). ESI-MS m/z 465.19[M+H].sup.+.
Example 83: Synthesis of Compound 83
[0347] ##STR00178##
[0348] The Compound 83 was synthesized according to the synthesis of Compound 81 by using Compound 83-1 to replace the Compound 5-1 in Example 81.
[0349] .sup.1H NMR (500 MHz, Chloroform) δ 10.96 (s, 1H), 9.72 (s, 1H), 7.86 (s, 1H), 7.43 (d, J=5.0 Hz, 1H), 7.39-7.26 (m, 2H), 7.14-6.95 (m, 5H), 5.04 (t, J=5.4 Hz, 1H), 4.84 (q, J=6.0 Hz, 1H), 3.69 (dt, J=12.7, 6.5 Hz, 1H), 3.56 (dt, J=12.7, 6.5 Hz, 1H), 3.29 (dd, J=12.4, 5.3 Hz, 1H), 3.04 (dd, J=12.4, 5.3 Hz, 1H), 2.61 (tt, J=9.1, 6.5 Hz, 1H), 2.23 (ddt, J=12.9, 9.1, 6.4 Hz, 1H), 2.02 (t, J=6.2 Hz, 2H), 1.99-1.88 (in, 1H). ESI-MS m/z 501.17[M+H].sup.+.
Example 84: Synthesis of Compound 84
[0350] ##STR00179##
[0351] The Compound 84 was synthesized according to the synthesis of Compound 47 by using Compound 84-1 to replace the Compound 14-1 in Example 47.
[0352] .sup.1H NMR (500 MHz, Chloroform) δ 11.97 (s, 1H), 9.72 (s, 1H), 7.50 (dd, J=14.9, 3.2 Hz, 1H), 7.11 (s, 1H), 7.03 (t, J=15.0 Hz, 1H), 6.85 (dd, J=15.0, 3.1 Hz, 1H), 6.45 (s, 2H), 5.09 (dd, J=25.7, 13.2 Hz, 1H), 4.46 (t, J=11.8 Hz, 1H), 3.74-3.48 (m, 2H), 2.66-2.53 (m, 4H), 2.21 (ddd, J=25.3, 16.7, 12.5 Hz, 1H), 2.07-1.88 (m, 4H), 1.86-1.63 (m, 7H), 1.31 (p, J=11.6 Hz, 2H), 1.17-0.97 (in, 3H). ESI-MS m/z 467 0.26 [M+H].sup.+.
Example 85: Synthesis of Compound 85
[0353] ##STR00180##
[0354] The Compound 85 was synthesized according to the synthesis of Compound 12 by using Compound 85-1 to replace the Compound 12-1 in Example 12.
[0355] .sup.1H NMR (500 MHz, Chloroform) 611.30 (s, 1H), 9.72 (s, 1H), 8.78 (s, 1H), 7.98 (dt, J=14.9, 3.1 Hz, 1H), 7.60 (dtd, J=14.7, 8.0, 3.2 Hz, 2H), 7.29-6.93 (m, 6H), 6.35 (s, 1H), 6.02 (s, 1H), 5.66 (s, 1H), 5.36 (dd, J=23.9, 11.7 Hz, 1H), 4.96 (t, J=10.8 Hz, 1H), 3.72-3.47 (m, 2H), 3.29 (dd, J=24.7, 10.8 Hz, 1H), 3.04 (dd, J=24.7, 10.8 Hz, 1H), 2.88 (tt, J=18.3, 12.5 Hz, 1H), 2.23 (ddt, J=25.5, 18.3, 12.8 Hz, 1H), 2.07-1.82 (in, 3H). ESI-MS m/z 465.19[M+H].sup.+.
Example 86: Synthesis of Compound 86
[0356] ##STR00181##
[0357] The Compound 86 was synthesized according to the synthesis of Compound 81 by using Compound 57-1 to replace the Compound 5-1 in Example 81.
[0358] .sup.1H NMR (500 MHz, Chloroform) δ 13.79 (s, 1H), 9.71 (s, 1H), 7.59 (dt, J=14.9, 7.4 Hz, 2H), 7.36-7.21 (m, 3H), 7.07-6.94 (m, 3H), 6.35 (s, 1H), 6.15 (s, 1H), 5.41 (s, 1H), 5.15 (dt, J=12.4, 10.3 Hz, 1H), 4.87 (t, J=14.7 Hz, 1H), 3.75-3.60 (m, 1H), 3.60-3.45 (m, 1H), 3.29 (dd, J=24.9, 14.5 Hz, 1H), 3.04 (dd, J=24.9, 14.5 Hz, 1H), 2.69 (tt, J=18.3, 6.8 Hz, 1H), 2.28-2.10 (m, 1H), 2.06-1.89 (in, 3H). ESI-MS m/z 466.18[M+H].sup.+.
Example 87: Synthesis of Compound 87
[0359] ##STR00182##
[0360] The Compound 87 was synthesized according to the synthesis of Compound 81 by using Compound 10-1 to replace the Compound 5-1 in Example 81.
[0361] .sup.1H NMR (500 MHz, Chloroform) δ 9.72 (s, 1H), 8.04 (s, 1H), 7.63-7.55 (m, 1H), 7.50 (dd, J=14.3, 3.6 Hz, 1H), 7.36-7.15 (m, 5H), 7.06-6.94 (m, 2H), 6.06 (s, 1H), 5.65 (td, J=15.4, 12.4 Hz, 1H), 5.55 (s, 1H), 4.90 (t, J=11.8 Hz, 1H), 3.70-3.44 (m, 2H), 3.29 (dd, J=24.8, 11.8 Hz, 1H), 3.04 (dd, J=24.8, 11.8 Hz, 1H), 2.54 (td, J=31.2, 16.3 Hz, 1H), 2.20 (ddt, J=25.0, 16.3, 13.0 Hz, 1H), 2.07-1.84 (in, 3H). ESI-MS m/z 466.18[M+H].sup.+.
Example 88: Synthesis of Compound 88
[0362] ##STR00183##
[0363] The Compound 88 was synthesized according to the synthesis of Compound 81 by using Compound 11-1 to replace the Compound 5-1 in Example 81.
[0364] .sup.1H NMR (500 MHz, Chloroform) δ 12.09 (s, 1H), 9.72 (s, 1H), 8.59 (dd, J=7.5, 1.5 Hz, 1H), 8.49 (s, 1H), 8.20 (dd, J=7.4, 1.3 Hz, 1H), 7.93-7.82 (m, 2H), 7.76 (d, J=7.5 Hz, 1H), 7.57 (td, J=7.5, 1.5 Hz, 1H), 7.39-7.26 (m, 2H), 7.00 (tdd, J=5.6, 2.9, 1.6 Hz, 2H), 6.10 (s, 1H), 5.23 (t, J=7.0 Hz, 1H), 4.55 (q, J=6.5 Hz, 1H), 3.65 (dt, J=12.8, 6.5 Hz, 1H), 3.52 (dt, J=12.6, 6.5 Hz, 1H), 3.29 (dd, J=12.5, 7.0 Hz, 1H), 3.04 (dd, J=12.5, 7.0 Hz, 1H), 2.44 (tt, J=9.1, 6.1 Hz, 1H), 2.21 (ddt, J=12.8, 9.1, 6.5 Hz, 1H), 2.02 (t, J=6.4 Hz, 2H), 1.92 (ddt, J=12.7, 9.3, 6.5 Hz, 1H). ESI-MS m/z 477.19[M+H].sup.+.
Test Example 1: Evaluation of Inhibitory Activity to 3CL Protease of 2019 Novel Coronavirus
[0365] Determination of the inhibitory activity of the compound against the 3CL protease of 2019 Novel Coronavirus (2019-nCoV 3CL.sup.pro): Fluorescence resonance energy transfer (FRET) technology was used to determine the enzyme level inhibitory activity of the 3C protease inhibitor. In a 96-well plate, 27.5 μL of buffer (20 mM Tris, 100 mM NaCl, 1 mM EDTA, pH 7.4) was added to each well, and 2.5 μL of compound (final concentrations 2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 12 μM, 14 μM, 16 μM, 18 μM, 20 μM) and 5 μL of EV713Cpro (final concentration 3 μM) were added together. The system was incubated for 15 min at 37° C., and then 15 μL of fluorescent substrate diluted in buffer (of which the final concentration was 20 μM) was added. The fluorescence parameters were measured with a Ge n5 fluorometer at 340 nm excitation wavelength and 490 nm emission wavelength, respectively. The system was kept at 37° C. for 10 min before the data was read. A negative control without any compound was used (the rest are the same). The obtained data was processed using GraphPad Prism 5 software, and the experimental results are shown in Tables 1A and 1B.
TABLE-US-00002 TABLE 1A Inhibitory Activity of 3CL protease of 2019 Novel Coronavirus Compound (1 μM) number Inhibition rate 5(DC402265) 95.7 6(DC402264) 95.8 7 88.7 8 86.5 9(DC402240) 95.8 10(DC402237) 88.2 11(DC402238) 89.7 12 103.1 16(DC402267) 89.1 (IC.sub.50 = 74 nM)) 17(DC402266) 105.7 18(DC402306) 97.8 19(DC402308) 98.3 20(DC402310) 100.7 24(DC402307) 93.3 25(DC402309) 101.9 26(DC402311) 101.8 35 94.3 45 85.2 46 89.3 47 90.4 48(DC402234) 100.4 49 91.2 50 89.1 51 84.6 56 81.3 57 92.3 58 94.3 59 86.7 60 92.6 61(DC402207) 88.3 65 86.7 66 68.4 81(DC402259) 96.3 82 100.7 83 97.8
[0366] Note: Compound 48, or DC402234, or 2234 (“DC40” in “DC40XXXX” was omitted) referred to the same compound; other compounds were deduced by analogy.
[0367] The IC.sub.50 values of some preferred compounds were listed in Table 1B.
TABLE-US-00003 TABLE IB Inhibition Activity(IC.sub.50) of 2019 n-Cov 3CL Protease Inhibitory Compound IC.sub.50 (nM) of SARS- number CoV-2 3 CL protease 5 33.61 ± 3.45 6 48.53 ± 3.17 9 68.14 ± 12.48 10 67.46 ± 13.88 11 178.90 ± 29.39 12 87 ± 8 16 74 17 61.03 ± 12.50 18 238.33 ± 18.67 19 62.73 ± 8.97 20 150.10 ± 9.55 24 131.83 ± 14.73 25 169.23 ± 37.87 26 105.29 ± 10.12 48 53 ± 0.5 81 40 ± 0.2 82 103 ± 13 83 113 ± 9
[0368] The experimental results show that most compounds have good inhibitory activity against 2019-nCoV 3CL protease at 1 μM, and IC.sub.50 values of some compounds (Compounds 48, 81, 5, 6, 9, 10, 12, 16, 17, 19) were less than 100 nM, in which the inhibitory activity (IC.sub.50) of Compound 16 (DC402267) against 2019-nCoV 3CL protease reached 73.5 nM, and the IC.sub.50 of Compound 48 and 81 reached 53 nM and 40 nM.
Test Example 2: Evaluation of the Inhibitory Activity of Compounds Against the Replication of 2019 n-Cov and Determination of Half Toxic Concentration Thereof
[0369] 2.1 EC.sub.50 Determination
[0370] Determination of the compound's inhibitory activity against the replication of 2019-nCov: 100 μl/well compounds at gradient concentration was added to 96 wells, then 50 μl/well of virus buffer was added, then 50 μl/well of cultured RD cells (rhabdomyosarcoma cells) was immediately added. The system was cultured at 37° C. for 3-4 days until the maximum cytopathic effect was observed. The medium was aspirated, 75 μl 5% MTS phenol red medium was added, and incubated at 37° C. with 5% CO.sub.2 for 1.5 hours. The fluorescence value of each well was measured at 498 nM wavelength, and a graph of compound concentration vs. cell response was drawn, and the EC.sub.50 of the compound against the virus was calculated using customized software purchased from Accelrys Corporation.
[0371] The test results are shown in
[0372] The results show that the aldehyde compounds of the present invention can effectively inhibit the replication of the 2019 n-Cov (
[0373] The existing anti-2019-nCoV positive compound CQ has an inhibitory rate against 2019-nCoV virus replication at EC.sub.50=1.13 μM. The results showed that: when using CQ as a positive control to test the compounds of the application at different concentration gradients, all of Compound 2234 (DC402234, namely Compound 48), 2259 (DC402259, namely Compound 81), 2267 (DC402267, namely Compound 16) have shown excellent anti-virus activity, in which the EC.sub.50 of 2234 was 0.29 μM, and the EC.sub.50 of 2259 was 0.33±0.09 μM, respectively. Therefore, the inhibitory rate of Compounds 48 and 81 to 2019-nCoV at viral level was better than that of the positive control CQ, thus showing good anti-2019-nCoV potentiality (
[0374] 2.2 Determination of Half-Toxic Concentration
[0375] In this example, the half-toxic concentrations (CC.sub.50) of some compounds of the present invention to Vero E6 cells were determined with CCK8 kit in duplicate. The results were shown in
Activity Example 3: Crystal Complex Formed by Compound and SARS-CoV-2 3CL.SUP.pro
[0376] 3.1 Cloning, Expression, Purification and Crystallization of SARS-CoV-2) 3CL.sup.pro (M.sup.pro)
[0377] The full-length gene encoding SARS-CoV-2 3CL.sup.pro was optimized and synthesized, and inserted into the BamHI and XhoI sites of pGEX-6p-1 plasmid DNA (Amersham Biosciences) for Escherichia coli (E. coli) Expression (GENEWIZ). SARS-CoV-2 3CL.sup.pro was further purified and then co-crystallized. SARS-CoV-2 3CL.sup.pro was incubated with 10 mM of Compound 48 or Compound 81 for 30 mins, and the crystallized by pendant drop vapor diffusion method at 20° C. (5 mg/ml). The best crystals were grown with buffer containing 2% polyethylene glycol (PEG) 6000, 3% DMSO, 1 mM DTT, 0.1 M MES (pH 6.0). The cryoprotectant solution contained 30% PEG 400, 0.1 M MES (pH 6.0).
[0378] 3.2 Data Collection and Refinement Statistics of Crystal Complexes
[0379] All data was collected on beamline BL19U1 of Shanghai Synchrotron Radiation Facility (SSRF) using Pilatus3 6M image plate detector at wavelength 0.9785 Å and 100 K. XDS25 program was used for data integration and expansion. SARS-CoV 3CL.sup.pro (PDB: 2H2Z) was used as the search model and the PHASER26 program was used for molecular substitution (MR) to determine the structure. The output model of MR was manually adjusted and iterative looped by Coot27, and refined with Phenix. Compounds 48 and 81 were constructed according to the outline.
[0380] 3.3 Results
[0381] The results are shown in Table 2 and
TABLE-US-00004 TABLE 2 Data collection and refinement statistics of crystal complexes Mpro-48 Mpro-81 PDB code 6LZE 6M0K Data collection Space group C2 C2 Unit cell size a, b, c (Å) 97.70, 80.94, 51.74 98.15, 81.70, 51.67 α, β, γ (°) 90, 114.27, 90 90, 114.69, 90 Wavelength (Å) 0.97852 0.97852 Resolution (Å) 50.00-1.51 (1.54-1.51).sup.a 50.00-1.50 (1.54-1.50).sup.a Completeness (%) 98.0 (91.6) 98.8 (90.3) R.sub.merge (%) 4.2 (55.5) 3.0 (77.1) Redundancy 3.3 (2.7) 3.4 (2.8) I/σ(I) 13.99 (1.80) 18.69 (1.30) Refinement Resolution (Å) 47.16-1.50 43.45-1.50 Number of reflections 57,378 58,412 R.sub.work/R.sub.free (%) 17.8/20.1 18.34/19.66 Number of atoms Protein 2340 2,347 Ligand 49 50 Water 209 163 B factor (Å.sup.2) Protein 28.75 31.92 Ligand 37.60 52.56 Water 37.95 40.62 R.m.s deviation Bond length (Å) 0.014 0.017 Bond angle (°) 1.280 1.440 Ramachandran graph (%) Favored 98.0 98.0 Allowed 2.0 2.0 Outliers 0.0 0.0 .sup.aThe value in parentheses was for the highest resolution structure.
[0382] The coordinates and structure factors of SARS-CoV-2 3CL.sup.pro and Compounds 48 and 81 have been stored in the protein database, of which the PDB numbers were 6LZE and 6M0K, respectively.
[0383] The structures of crystal complexes of Compounds 48, 81 and SARS-CoV-2 3CL.sup.pro were shown in
[0384] The experimental results showed that the aldehyde group C of Compound 48 formed a standard 1.8 Å C—S covalent bond with the catalytic site Cys145 of SARS-CoV-2 MP″ (
[0385] The crystal structure of Compound 81 and SARS-CoV-2 M″ was very similar that of Compound 48, which shows similar inhibitor binding modes (
[0386] In short, these two crystal structures revealed a same inhibitory mechanism, that is, the two compounds occupying the substrate binding pocket, mimicking the intermediate in the catalytic reaction, thus blocking the enzyme activity of SARS-CoV-2 M.sup.pro.
[0387] All literatures mentioned in the present invention are incorporated herein by reference, as though each one is individually incorporated by reference. Additionally, it should be understood that after reading the above teachings, those skilled in the art can make various changes and modifications to the present invention. These equivalents also fall within the scope defined by the appended claims.