POLYARYL CARBOXYLIC FULLERENE DERIVATIVE AND USE THEREOF IN ANTI-CORONAVIRUS INFECTION

Abstract

A polyaryl carboxylic fullerene derivative and its use in anti-coronavirus infection are disclosed. Specifically, a compound shown in formula A: fullerene-RR.sub.1R.sub.2R.sub.3R.sub.4R.sub.5 formula A, or a stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound is provided. The compound or a derivative thereof has promising application prospects in anti-coronavirus infection.

Claims

1. A compound of a general formula shown in formula A: ##STR00252## or a stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound, wherein fullerene in the formula A is a cage-like all-carbon structure comprising a five-membered carbon ring, a six-membered carbon ring, and/or a seven-membered carbon ring and/or a four-membered carbon ring, or the fullerene is a metal or cluster-embedded structure; the fullerene is selected from the group consisting of C.sub.60, C.sub.70, C.sub.84, C.sub.90, and C.sub.100 wherein R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy, ; (2) ##STR00253## wherein Y .sup.1, Y.sup.2, Y.sup.3, Y.sup.4, and Y.sup.5 are not all H and each are independently selected from the group consisting of the following: 1) H, and 2) ##STR00254## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; and (3) ##STR00255## wherein Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 are not all H; R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 each are independently selected from the group consisting of the following: (1) ##STR00256## wherein Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 are not all H; (2) ##STR00257## wherein Y .sup.1, Y.sup.2, Y.sup.3, Y.sup.4, and Y.sup.5 are not all H and each are independently selected from the group consisting of the following: 1) H, and 2) ##STR00258## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; the compound shown in formula A does not comprise the following compounds: TABLE-US-00007 .

2. The compound according to claim 1, wherein R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy ; and (2) ##STR00277## wherein Y.sup.3 is ##STR00278## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00279## wherein X.sup.3 is C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups; or R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 each are independently selected from the group consisting of the following: (1) ##STR00280## wherein Z.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups ; and (2) ##STR00281## wherein Y.sup.3 is ##STR00282## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00283## wherein X.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups.

3. The compound according to claim 1, wherein the compound is a compound selected from the group consisting of the following: TABLE-US-00008 compound No. compound structure 3 TABLE-US-00009 5 6 or a stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound 3, the compound 4, the compound 5, or the compound 6.

4. A method for preparing the compound according to claim 3, comprising: (1) subjecting C.sub.60Cl.sub.6 to a nucleophilic substitution reaction with α-methylhydrocinnamic acid, 5-phenylvaleric acid, or 3-(4-biphenyl)propionic acid to obtain the compound; or subjecting C.sub.60Cl.sub.6 to a nucleophilic substitution reaction with methyl 3-(4-biphenyl)propionate to obtain a first resulting reaction system, and after the nucleophilic substitution reaction is complete, cooling the first resulting reaction system to room temperature, and subjecting a product of the nucleophilic substitution reaction to a hydrolysis reaction to obtain the compound, wherein a molar ratio of the α-methylhydrocinnamic acid, the 5-phenylvaleric acid, the methyl 3-(4-biphenyl)propionate, or the 3-(4-biphenyl)propionic acid to the C.sub.60Cl.sub.6 is (20-30):1; the nucleophilic substitution reaction is conducted in the presence of SnCl.sub.4; the nucleophilic substitution reaction is conducted under water-free and oxygen-free conditions; a solvent for the nucleophilic substitution reaction is nitrobenzene; the nucleophilic substitution reaction is conducted at 60° C. to 100° C. for 1 h to 3 h; a solvent for the hydrolysis reaction is toluene; the hydrolysis reaction is conducted in the presence of acetic acid and hydrochloric acid; the hydrolysis reaction is conducted at 60° C. to 100° C. for 60 h to 80 h; before the hydrolysis reaction, the method further comprises: subjecting the product of the nucleophilic substitution reaction to purification, wherein the purification is conducted through column chromatography and a mobile phase for the column chromatography is toluene/methanol in a volume ratio of 85/15; and (2) after the nucleophilic substitution reaction of the C.sub.60Cl.sub.6 with the α-methylhydrocinnamic acid, the 5-phenylvaleric acid, or the 3-(4-biphenyl)propionic acid in step (1) is complete, cooling a second resulting reaction system to room temperature, adding acetonitrile to the second resulting reaction system for precipitation to obtain a resulting mixture, and subjecting a the resulting mixture to filtration to obtain a first filter cake; or after the hydrolysis reaction in step (1) is complete, conducting extraction with toluene, rotary evaporation, adding acetonitrile to a reaction product obtained after the rotary evaporation for precipitation to obtain a resulting mixture, and subjecting the resulting mixture to filtration to obtain a first filter cake; (3) dissolving the first filter cake with a potassium hydroxide solution to obtain a resulting solution, subjecting the resulting solution to filtration to remove insoluble matters, and collecting a filtrate; (4) adding hydrochloric acid dropwise to the filtrate for neutralization until a pH of the filtrate is 7.0, such that a precipitate is produced; and (5) subjecting a mixture obtained in step (4) to filtration to obtain a second filter cake, wherein the second filter cake is the compound .

5. A pharmaceutical composition comprising the compound or the stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound according to claims 1 and an optional pharmaceutically acceptable carrier or excipient, wherein the pharmaceutical composition is in a dosage form of a pill, a tablet, a capsule, an aqueous solution, or an aqueous suspension for an oral administration, in a dosage form of a micronized suspension or solution for a topical ocular administration, in a dosage form of a paste, an ointment, a lotion, a spray, or a cream for a topical transdermal or mucosal administration, or in a dosage form of sterile injection water, an oil suspension, or a sterile injection solution for an injection.

6. A vesicle with a diameter of 40 nm to 140 nm, wherein the vesicle is produced using the compound or the pharmaceutically acceptable salt thereof the compound according to claims 1; and the vesicle is produced by a method comprising: dissolving the compound or the pharmaceutically acceptable salt thereof the compound in N,N-dimethylformamide (DMF) or acetonitrile to obtain a solution, and subjecting the solution to an ultrasonic treatment at 30° C. to 40° C. for 1 h to 2 h to obtain the vesicle, wherein a ratio of a mass of the compound or the pharmaceutically acceptable salt thereof the compound to a volume of the DMF or the acetonitrile is 1 mg : 1 mL.

7. A method of a use of a compound shown in formula A or a stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate thereof the compound in a preparation of a drug, wherein the drug is provided for preventing and/or treating a disease caused by a coronavirus infection, the disease is a respiratory disease comprising a simple infection, pneumonia, an acute or severe acute respiratory infection, hypoxemic respiratory failure (HRF) and acute respiratory distress syndrome (ARDS), sepsis, and septic shock, pneumonia, the pneumonia is pneumonia COVID-2019, the simple infection comprises fever, cough, and sore throat ; or the drug is provided to serve as an inhibitor for a coronavirus ; or the drug is provided to inhibit a replication or a reproduction of a coronavirus in a cel, ##STR00287## wherein fullerene in the formula A is a cage-like all-carbon structure comprising a five-membered carbon ring, a six-membered carbon ring, and/or a seven-membered carbon ring and/or a four-membered carbon ring, or the fullerene is a metal or cluster-embedded structure; the fullerene is selected from the group consisting of C.sub.60, C.sub.70, C.sub.84, C.sub.90, and C.sub.100; wherein R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy ; (2) ##STR00288## wherein Y .sup.1, Y.sup.2, Y.sup.3, Y.sup.4, and Y.sup.5 are not all H and each are independently selected from the group consisting of the following: 1) H, and 2) ##STR00289## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; and (3) ##STR00290## wherein Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 are not all H; R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 each are independently selected from the group consisting of the following: (1) ##STR00291## wherein Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 are not all H; and (2) ##STR00292## wherein Y .sup.1, Y.sup.2, Y.sup.3, Y.sup.4, and Y.sup.5 are not all H and each are independently selected from the group consisting of the following: 1) H, and 2) ##STR00293## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H.

8. The method of the use according to claim 7, wherein R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy, and (2) ##STR00294## wherein Y.sup.3 is ##STR00295## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00296## wherein X.sup.3 is C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, or R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 each are independently selected from the group consisting of the following: (1) ##STR00297## wherein Z.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, ; and (2) ##STR00298## wherein Y.sup.3 is ##STR00299## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00300## wherein X.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups.

9. The method of the use according to claim 7, wherein the compound shown in formula A is selected from the group consisting of the following: TABLE-US-00010 compound No. compound structure 1 2 3 4 5 6 .

10. A method of a use of a pharmaceutical composition or a vesicle in a preparation of a drug, wherein the pharmaceutical composition comprises the compound shown in formula A or the stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound according to claim 7 and an optional pharmaceutically acceptable carrier or excipient; the vesicle has a diameter of 40 nm to 140 nm and is produced using the compound shown in formula A or the pharmaceutically acceptable salt thereof the compound ; the drug is provided for preventing and/or treating a disease caused by a coronavirus infection, the disease is a respiratory disease comprising a simple infection, pneumonia, an acute or severe acute respiratory infection, hypoxemic respiratory failure (HRF) and acute respiratory distress syndrome (ARDS), sepsis, and septic shock, the pneumonia is pneumonia COVID-2019 the simple infection comprises fever, cough, and sore throat ; or the drug is provided to serve as an inhibitor for a coronavirus ; or the drug is provided to inhibit a replication or a reproduction of a coronavirus ; in a cell the pharmaceutical composition is in a dosage form of a pill, a tablet, a capsule, an aqueous solution, or an aqueous suspension for an oral administration, in a dosage form of a micronized suspension or solution for a topical ocular administration, in a dosage form of a paste, an ointment, a lotion, a spray, or a cream for a topical transdermal or mucosal administration, or in a dosage form of sterile injection water, an oil suspension, or a sterile injection solution for an injection; and the vesicle is produced by a method comprising: dissolving the compound shown in formula A or the pharmaceutically acceptable salt of the compound in DMF or acetonitrile to obtain a solution, and subjecting the solution to an ultrasonic treatment at 30° C. to 40° C. for 1 h to 2 h to obtain the vesicle, wherein a ratio of a mass of the compound shown in formula A or the pharmaceutically acceptable salt thereof the compound to a volume of the DMF or the acetonitrile is 1 mg : 1 mL.

11. The compound according to claim 1, wherein the fullerene is C.sub.60 having a general formula shown in formula I, and the fullerene is a hollow cage-like structure; ##STR00307##

12. The compound accorrding to claim 1, wherein R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy; and (2) ##STR00308## wherein Y .sup.1, Y.sup.2, Y.sup.3, Y.sup.4, and Y.sup.5 are not all H and each are independently selected from the group consisting of the following: 1) H, and 2) ##STR00309## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H.

13. The compound according to claim 2, wherein the compound is a compound selected from the group consisting of the following: TABLE-US-00011 compound No. compound structure 3 4 5 6 or a stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound 3, the compound 4, the compound 5, or the compound 6.

14. The method according to claim 4, wherein after step (2) and before step (3), the method further comprises: washing and drying the first filter cake; and after step (5), the method further comprises: drying the second filter cake to obtain the compound.

15. The pharmaceutical composition according to claim 5, wherein in the compound, R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy; and (2) ##STR00314## wherein Y.sup.3 is ##STR00315## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00316## wherein X.sup.3 is C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups; or R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 each are independently selected from the group consisting of the following: (1) ##STR00317## wherein Z.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups; and (2) ##STR00318## wherein Y.sup.3 is ##STR00319## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00320## wherein X.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups.

16. The pharmaceutical composition according to claim 5, wherein the compound is a compound selected from the group consisting of the following: TABLE-US-00012 compound No. compound structure 3 4 5 6 or a stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound 3, the compound 4, the compound 5, or the compound 6.

17. The vesicle according to cliam 6, wherein in the compound, R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy; and (2) ##STR00325## wherein Y.sup.3 is ##STR00326## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00327## wherein X.sup.3 is C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups; or R.sup.1, R.sup.2, R.sup.3, R.sup.4, and R.sup.5 each are independently selected from the group consisting of the following: (1) ##STR00328## wherein Z.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups; and (2) ##STR00329## wherein Y.sup.3 is ##STR00330## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H; or, Y.sup.3 is ##STR00331## wherein X.sup.3 is independently C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups.

18. The vesicle according to cliam 6, wherein the compound is a compound selected from the group consisting of the following: TABLE-US-00013 compound No. compound structure 3 4 TABLE-US-00014 5 6 or a stereoisomer, prodrug, pharmaceutically acceptable salt, or pharmaceutically acceptable solvate of the compound 3, the compound 4, the compound 5, or the compound 6.

19. The method of the use according to claim 7, wherein the fullerene is C.sub.60 having a general formula shown in formula I, and the fullerene is a hollow cage-like structure; ##STR00336##

20. The method of the use according to claim 7, wherein R is selected from the group consisting of the following: (1) H, F, Cl, Br, I, C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy; and (2) ##STR00337## , wherein Y .sup.1, Y.sup.2, Y.sup.3, Y.sup.4, and Y.sup.5 are not all H and each are independently selected from the group consisting of the following: 1) H, and 2) ##STR00338## wherein X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 each are independently selected from the group consisting of H and C.sub.1-C.sub.6 alkyl substituted with 1, 2, or 3 carboxyl groups, and X.sup.1, X.sup.2, X.sup.3, X.sup.4, and X.sup.5 are not all H.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0404] FIG. 1 is a schematic diagram illustrating the structure of the vesicle of the present disclosure;

[0405] FIG. 2 shows the survival results of African green monkey kidney (AGMK) cells in the presence of a potassium salt of a polyaryl carboxylic fullerene derivative 4 (compound 4) in an embodiment of the present disclosure at a concentration of 109.2 .Math.M;

[0406] FIGS. 3A-3C show the inhibition results of a potassium salt of a polyaryl carboxylic fullerene derivative 4 (compound 4) in an embodiment of the present disclosure on a novel coronavirus (SARS-CoV-2) at a concentration of 54.6 .Math.M (3 replicates);

[0407] FIG. 4 shows the control results of AGMK cells in the absence of a viral infection and a polyaryl carboxylic fullerene derivative 4 (compound 4) according to an embodiment of the present disclosure;

[0408] FIG. 5 shows the control results of novel coronavirus (SARS-CoV-2)-infected AGMK cells in the absence of a polyaryl carboxylic fullerene derivative 4 (compound 4) according to an embodiment of the present disclosure;

[0409] FIGS. 6A-6B show scanning electron microscopy (SEM) images of a vesicle according to an embodiment of the present disclosure;

[0410] FIGS. 7A-7B show SEM images of a vesicle according to another embodiment of the present disclosure; and

[0411] FIGS. 8A-8B show SEM images of a vesicle according to another embodiment of the present disclosure.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0412] The embodiments of the present disclosure are described in detail below through specific examples, but in any case, the examples should not be interpreted as limitations on the present disclosure.

[0413] An objective of the present disclosure is to provide a water-soluble polyaryl carboxylic fullerene derivative and a use thereof in the preparation of a drug for preventing or treating a coronavirus.

[0414] In particular, to solve the technical problem of the present disclosure, the following technical solution is adopted:

##STR00245##

where n is any integer from 1 to 6.

[0415] In some embodiments, the coronavirus is not limited to the novel coronavirus (SARS-CoV-2).

[0416] In some embodiments, the drug is a drug with the water-soluble polyaryl carboxylic fullerene derivative shown in formula (O) as an active ingredient.

[0417] In some embodiments, the drug is a monomer compound with the water-soluble polyaryl carboxylic fullerene derivative shown in formula (O) as an active ingredient.

[0418] In some embodiments, the drug includes the water-soluble polyaryl carboxylic fullerene derivative shown in formula (O) and a pharmaceutically acceptable carrier or excipient.

[0419] In some embodiments, the drug has a dosage form of a pill, a tablet, a capsule, or an oral liquid.

[0420] The present disclosure is further explained below in conjunction with specific examples.

[0421] 6 water-soluble polyaryl carboxylic fullerene derivatives (compounds 1 to 6) and potassium salts thereof are synthesized by the inventors, and a role of the potassium salts in the inhibition of a coronavirus in vitro is evaluated. A BHK21-hACE2 cell and an rVSV-SARS2 pseudocoronavirus were used to conduct an in vitro experiment, and an AGMK cell (Vero E6) and a novel coronavirus (SARS-CoV-2) were also used to conduct an in vitro experiment, such as to screen candidate compounds.

Example 1 Preparation of Compounds

[0422] Specific molecular structures of the 6 water-soluble polyaryl carboxylic fullerene derivatives (compounds 1 to 6) synthesized by the inventors were shown in Table 1 below.

TABLE-US-00005 Compound structures Compound No. Compound structure 1 [00246] 2 [00247] 3 [00248] 4 [00249] 5 [00250] 6 [00251]

[0423] 1. Preparation methods of compounds 1 and 2 can be found in Org. Biomol. Chem., 2019, 17, 7155-7160.

[0424] 2. A preparation method for compound 3 is as follows:

[0425] C.sub.60Cl.sub.6 (200 mg, 0.214 mmol, 1 equiv) and α-methylhydrocinnamic acid (878 mg, 5.36 mmol, 25 equiv) were dissolved in 50 mL of nitrobenzene under water-free and oxygen-free conditions, then 0.1 mL of SnCl.sub.4 was added, and a reaction was conducted at 80° C. for 2 h. After the reaction was complete, the resulting system was cooled to room temperature, and 300 mL of acetonitrile was added for precipitation. The resulting precipitate was filtered out through suction filtration, washed with n-hexane three times, dried, and then dissolved with a potassium hydroxide solution. The resulting solution was filtered to remove insoluble matters, a filtrate was collected, and hydrochloric acid was added dropwise to the filtrate until a pH of the filtrate was 7.0, such that a red precipitate was produced. The red precipitate was filtered out through suction filtration and then dried to obtain a product 3 (compound 3).

[0426] Identification of compound 3:

[0427] .sup.1H NMR (500 MHz, DMSO-d6, δ, ppm) 12.14 (br.s, 5H), 7.80 (m, 4H), 7.53 (m, 4H), 7.36-7.14 (m, 6H), 7.11-6.97 (m, 4H), 6.85-6.61 (m, 2H), 2.96 (m, 4H), 2.64 (m, 1H), 2.42-1.89 (m, 5H), 1.24 (m, 5H), 1.02 (m, 12H).

[0428] .sup.13CNMR (126 MHz, DMSO-d.sub.6, δ, ppm) 176.27, 148.81, 148.76, 148.68, 148.68, 148.49, 148.39, 148.31, 148.25, 148.08, 148.08, 147.95, 147.87, 147.64, 147.29, 144.54, 144.42, 144.41, 144.24, 144.14, 144.07, 144.04, 143.88, 143.69, 143.63, 139.58, 139.14, 139.02, 138.88, 138.86, 138.70, 138.54, 138.39, 138.07, 135.43, 135.20, 130.13, 129.96, 129.75, 129.47, 129.43, 129.38, 129.35, 129.28, 129.18, 129.18, 128.95, 128.90, 128.78, 128.69, 128.54, 128.43, 128.26, 128.20, 128.06, 128.03, 127.91, 127.83, 127.73, 127.52, 125.03, 77.27, 77.27, 77.22, 77.02, 76.76, 51.63, 41.28, 39.22, 32.21, 29.71, 29.33, 26.40, 23.43.

[0429] Molecular formula: C.sub.110H.sub.57ClO.sub.10; and mass spectrometry (MS) (MALDI-TOF), m/z: 1571.7.

[0430] 3. A preparation method for compound 4 is as follows:

[0431] C.sub.60Cl.sub.6 (200 mg, 0.214 mmol, 1 equiv) and 5-phenylvaleric acid (953 mg, 5.36 mmol, 25 equiv) were dissolved in 50 mL of nitrobenzene under water-free and oxygen-free conditions, then 0.1 mL of SnCl.sub.4 was added, and a reaction was conducted at 80° C. for 2 h. After the reaction was complete, the resulting system was cooled to room temperature, and 300 mL of acetonitrile was added for precipitation. The resulting precipitate was filtered out through suction filtration, washed with n-hexane three times, dried, and then dissolved with a potassium hydroxide solution; the resulting solution was filtered to remove insoluble matters, a filtrate was collected, and hydrochloric acid was added dropwise to the filtrate until a pH of the filtrate was 7.0, such that a red precipitate was produced. The red precipitate was filtered out through suction filtration and then dried to obtain a product 4 (compound 4).

[0432] Identification of compound 4:

[0433] .sup.1H NMR (500 MHz, DMSO-d.sub.6, δ, ppm) 11.96 (br.s, 5H), 7.72 (m, 1H), 7.54 (m, 3H), 7.47-7.24 (m, 3H), 7.13 (m, 6H), 7.00 (m, 2H), 6.85 (m, 3H), 6.67 (m, 2H), 2.57 (m, 3H), 2.46 (m, 4H), 2.21 (m, 8H), 2.15 (m, 3H), 1.55 (m, 4H), 1.48 (m, 14H), 1.23(m, 4H).

[0434] .sup.13CNMR (126 MHz, DMSO-d.sub.6, δ, ppm)δ 174.83, 174.78, 156.80, 156.66, 155.33, 154.68, 153.07, 151.92, 151.37, 150.95, 148.49, 148.43, 148.41, 148.37, 148.19, 148.03, 147.90, 147.88, 147.84, 147.68, 147.53, 147.36, 147.32, 147.18, 147.09, 147.03, 146.90, 146.90, 146.87, 146.82, 146.76, 146.73, 146.69, 146.58, 146.51, 146.40, 146.39, 146.15, 145.96, 145.93, 145.75, 137.28, 136.96, 136.83, 136.52, 129.86, 129.83, 129.72, 129.53, 129.38, 129.33, 129.22, 129.17, 129.05, 129.04, 128.79, 128.70, 128.51, 128.36, 128.17, 128.02, 127.65, 127.63, 127.63, 127.55, 127.38, 127.28, 127.04, 64.81, 61.06, 60.82, 58.69, 58.10, 57.04, 40.80, 40.52, 40.35, 40.19, 40.02, 39.85, 39.69, 39.52, 34.70, 34.01, 33.97, 30.68, 24.37, 24.32.

[0435] Molecular formula: C.sub.115H.sub.67ClO.sub.10; and MS (MALDI-TOF), m/z: 1641.7.

[0436] 4. A preparation method for compound 5 is as follows:

[0437] C.sub.60Cl.sub.6 (200 mg, 0.214 mmol, 1 equiv) and methyl 3-(4-biphenyl)propionate (1.286 g, 5.36 mmol, 25 equiv) were dissolved in 50 mL of nitrobenzene under water-free and oxygen-free conditions, then 0.1 mL of SnCl.sub.4 was added, and a reaction was conducted at 90° C. for 2 h. After the reaction was complete, the resulting system was cooled to room temperature, and the reaction product was directly separated through column chromatography with toluene/methanol in a volume ratio of 85/15 as a mobile phase. The solvent was removed through rotary evaporation, then 30 mL of toluene, 30 mL of acetic acid, and 5 mL of hydrochloric acid were added, and the reaction was conducted at 80° C. for 72 h. After the reaction was complete, an organic phase was subjected to extraction with toluene, and the toluene was removed through rotary evaporation, and 300 mL of acetonitrile was added for precipitation. The resulting precipitate was filtered out through suction filtration, washed with n-hexane three times, dried, and then dissolved with a potassium hydroxide solution. The resulting solution was subjected to suction filtration, a filtrate was collected, and hydrochloric acid was added dropwise to the filtrate until a pH of the filtrate was 7.0, such that a red precipitate was produced. The red precipitate was filtered out through suction filtration and then dried to obtain a product 5 (compound 5).

[0438] Identification of compound 5:

[0439] .sup.1H NMR (500 MHz, DMSO) δ 12.05 (br.s, 5H), 8.04-7.37(m 20H), 7.25(m, 5H), 7.18(m, 5H), 3.37(m, 10H), 2.81(m, 10H), 2.30(m, 5H), 1.23(m, 5H).

[0440] .sup.13C NMR (126 MHz, DMSO) δ 174.13, 174.11, 150.68, 150.23, 149.75, 149.59, 149.03, 148.81, 148.67, 148.34, 148.07, 147.81, 147.53, 147.35, 147.21, 146.69, 146.23, 145.70, 145.55, 145.15, 144.74, 144.59, 144.15, 143.85, 143.52, 143.16, 142.84, 142.61, 142.52, 142.08, 141.55, 141.15, 140.92, 140.79, 140.70, 140.63, 140.53, 140.45, 140.28, 140.13, 140.06, 139.68, 139.46, 139.30, 138.96, 138.57, 137.89, 137.80, 137.50, 136.48, 133.43, 132.63, 132.17, 130.58, 129.48, 129.35, 129.26, 129.11, 129.03, 128.94, 128.79, 128.65, 127.21, 127.07, 127.01, 126.85, 126.75, 126.70, 125.95, 125.76, 40.52, 40.35, 40.18, 40.02, 39.85, 39.68, 39.52, 37.82, 35.50, 30.42, 29.49, 26.33, 22.70, 21.51, 20.90.

[0441] Molecular formula: C.sub.133H.sub.65ClO.sub.10; and MS (MALDI-TOF), m/z: 1847.3

[0442] 5. A preparation method for compound 6 is as follows:

[0443] C.sub.60Cl.sub.6 (200 mg, 0.214 mmol, 1 equiv) and 3-(4-biphenyl)propionic acid (1.211 g, 5.36 mmol, 25 equiv) were dissolved in 50 mL of nitrobenzene under water-free and oxygen-free conditions, then 0.1 mL of SnCl.sub.4 was added, and a reaction was conducted at 90° C. for 2 h. After the reaction was complete, the resulting system was cooled to room temperature, and 300 mL of acetonitrile was added for precipitation. The resulting precipitate was filtered out through suction filtration, washed with n-hexane three times, dried, and then dissolved with a potassium hydroxide solution. The resulting solution was subjected to suction filtration, a filtrate was collected, and hydrochloric acid was added dropwise to the filtrate until a pH of the filtrate was 7.0, such that a red precipitate was produced. The red precipitate was filtered out through suction filtration and then dried to obtain a product 6 (compound 6).

[0444] Identification of compound 6:

[0445] .sup.1H NMR (500 MHz, DMSO) δ 12.08 (br.s, 6H), 8.14-7.47(m 24H), 7.35-6.78(m, 24H), 3.37(m, 12H), 2.77(m, 12H), 2.30-1.22(m, 12H).

[0446] .sup.13C NMR (126 MHz, DMSO) δ 174.14, 174.14, 174.12, 150.01, 149.36, 148.77, 148.51, 148.24, 147.58, 147.34, 147.24, 146.80, 146.13, 145.52, 145.05, 144.35, 144.03, 143.94, 143.61, 143.50, 142.84, 141.07, 140.99, 140.90, 140.77, 140.50, 140.48, 140.35, 140.23, 140.13, 139.98, 139.48, 138.46, 137.47, 137.35, 137.21, 137.02, 136.89, 136.79, 129.34, 129.25, 129.22, 129.14, 129.09, 129.06, 128.92, 127.98, 127.31, 127.17, 127.12, 127.07, 127.01, 126.94, 126.89, 126.83, 126.78, 126.73, 126.64, 126.49, 126.33, 126.16, 116.13, 40.48, 40.31, 40.14, 39.98, 39.81, 39.64, 39.48, 35.59, 35.49, 31.74, 30.38, 29.54, 29.48, 29.43, 29.32, 29.29, 29.04, 27.02.

[0447] Molecular formula: C.sub.150H.sub.78O.sub.12; and MS (MALDI-TOF), m/z: 2072.3

[0448] 6. A preparation method for potassium salts of compounds 1 to 6 is as follows:

[0449] The polyaryl carboxylic fullerene derivatives 1 to 6 (0.07 mmol, 1 equiv) each were added to distilled water (20 mL), anhydrous potassium carbonate (24.2 mg, 0.175 mmol, 2.5 eqiv) was added, and the resulting mixture was stirred until solids were completely dissolved. The resulting solution was then filtered through a PES syringe filter (with an average pore size of 0.45 .Math.m) and lyophilized for 8 h to finally obtain a powdered potassium salt.

Example 2 Toxicity Evaluation and Drug Activity Evaluation

1. Evaluation of Toxicity of the Water-Soluble Polyaryl Carboxylic Fullerene Derivative for Vero E6 cells

[0450] Method:

[0451] The potassium salt of the polyaryl carboxylic fullerene derivative 4 (namely, the potassium salt of compound 4) was completely dissolved in water to obtain an orange-red solution, and the orange-red solution was filtered to remove a small amount of an orange-red precipitate and then thoroughly shaken until the solution was clear. A monolayer of Vero E6 cells on a 96-well plate was washed once with phosphate-buffered saline (PBS), the resulting supernatant was discarded, and the potassium salt of the water-soluble polyaryl carboxylic fullerene derivative (namely, the potassium salt of compound 4) fold-diluted was added. In the normal cell group, an equal volume of a medium was added to each well. The cells were cultivated at 37° C. and 5% CO.sub.2 for 4 d to 5 d. A cytopathic effect (CPE) was observed and recorded under a microscope, and an inhibition rate was calculated. The medium was 10% fetal bovine serum (FBS)-containing DMEM.

[0452] Results:

[0453] The potassium salt of the polyaryl carboxylic fullerene derivative 4 (namely, the potassium salt of compound 4) exhibited no toxicity for AGMK cells (Vero E6) (as shown in FIG. 2) when at a concentration as high as 109.2 .Math.M.

2. Evaluation of Efficacy of the Water-Soluble Polyaryl Carboxylic Fullerene Derivative for A Novel Coronavirus (SARS-CoV-2)

[0454] Method:

Preventive Medication Model: CPE Reduction Assay

[0455] A monolayer of Vero E6 cells were washed once with PBS, the potassium salt of the polyaryl carboxylic fullerene derivative was added to each well at a specified concentration, and controls were set. The cells were incubated at 37° C. for 2 h, infected with a novel coronavirus (SARS-CoV-2) at a dose of about 100 TCID50, and incubated for 2 h. The used medium was replaced with a fresh medium, and the cells were further cultivated at 37° C. for 2 d. A fresh medium treated with the potassium salt of the polyaryl carboxylic fullerene derivative at a specified concentration (maintaining the concentration of the polyaryl carboxylic fullerene derivative) was used to further cultivate the cells for 2 d to 3 d. CPE was observed and recorded under a microscope, and a median effective concentration (IC.sub.50) was calculated by the Reed-Muench method.

[0456] Results:

[0457] The efficacy of the potassium salt of the water-soluble polyaryl carboxylic fullerene derivative 4 (namely, the potassium salt of compound 4) was evaluated by an AGMK cell (Vero) CPE inhibition method, and the median effective concentration (IC.sub.50) of the derivative for the novel coronavirus (SARS-CoV-2) was about 54.6 .Math.M (inhibition results of 3 parallel experiments were shown in FIGS. 3A-3C, and experimental results of the controls were shown in FIG. 4 and FIG. 5).

3. Evaluation of Toxicity of the Water-Soluble Polyaryl Carboxylic Fullerene Derivative for BHK21-hACE2 Cells

[0458] Method: The potassium salts of all water-soluble polyaryl carboxylic fullerene derivatives (namely, potassium salts of compounds 1 to 6) each were diluted to 1,000 .Math.M. Diluted analytes were then prepared by a serial double dilution method. 80 .Math.L of a diluted fullerene compound solution was mixed with 20 .Math.L of an rVSV-SARS-2 pseudovirus, the resulting mixture was added to pre-cultivated BHK21-hACE2 cells (80 .Math.L per well), and the cells were incubated for 12 h. The resulting supernatant was removed, then a mixture of 10 .Math.L of a cck-8 solution and 100 .Math.L of a medium was added, and the cells were cultivated for 2 h; the absorbance at 450 nm was determined. A reduction rate (%) of the absorbance was used to indicate the cytotoxicity of the compound.

[0459] Results:

[0460] The cytotoxicity experiments showed that the potassium salts of the water-soluble polyaryl carboxylic fullerene derivatives (namely, the potassium salts of compounds 1 to 6) exhibited no toxicity for BHK21-hACE2 cells when at a concentration as high as 1,000 .Math.M.

4. Evaluation of Efficacy of the Water-Soluble Polyaryl Carboxylic Fullerene Derivative for an rVSV-SARS2 Pseudocoronavirus

[0461] Method:

[0462] BHK21-hACE2 cells were plated on a 96-well plate at 2 × 10.sup.4 cells/well, such that a cell density could reach 70% to 80% after the cells were cultivated for 12 h. The potassium salts of the water-soluble polyaryl carboxylic fullerene derivatives (namely, the potassium salts of compounds 1 to 6, 2 mM) each were diluted as follows: 10 .Math.l of the potassium salt (2 mM) of the water-soluble polyaryl carboxylic fullerene derivative was mixed with 90 .Math.l of DMEM to obtain a sample solution with a final concentration of 0.2 mM, 50 .Math.l of the 0.2 mM sample solution was taken and mixed with 150 .Math.l of DMEM to obtain a sample solution of gradient 1, 50 .Math.l of the sample solution of gradient 1 was taken and mixed with 100 .Math.l of DMEM to obtain a sample solution of gradient 2, and 3-fold dilution was conducted in turn to obtain 8 gradients in total. 80 .Math.l of the potassium salt of the water-soluble polyaryl carboxylic fullerene derivative diluted at each gradient was taken and mixed with 20 .Math.l of an rVSV-SARS-2 (3 × 10e5 pfu/mL), and the resulting mixture was thoroughly mixed and then incubated at 37° C. for 1 h. A culture supernatant of BHK21-hACE2 cells was removed, and 100 .Math.l of a sample obtained after the incubation of 80 .Math.l of the water-soluble polyaryl carboxylic fullerene derivative and 20 .Math.l of rVSV-SARS-2 was added. 12 h later, the number of positive cells was recorded under a microscope, and CPE was recorded.

[0463] Results:

[0464] The efficacy of the potassium salts of the water-soluble polyaryl carboxylic fullerene derivatives was evaluated by a BHK21-hACE2 cell CPE inhibition method, and median effective concentrations (IC.sub.50) of the derivatives for the pseudocoronavirus (rVSV-SARS-2) were 79.6 .Math.M, 83.7 .Math.M, 189.3 .Math.M, 65.8 .Math.M, 42.2 .Math.M, and 8.5 .Math.M, respectively (as shown in Table 2).

TABLE-US-00006 Efficacy of the potassium salts of the water-soluble polyaryl carboxylic fullerene derivatives evaluated by the BHK21-hACE2 cell CPE inhibition method Compound 1 2 3 4 5 6 IC.sub.50/rVSV-SARS-2 (.Math.M) 79.6 83.7 189.3 65.8 42.2 8.5

Example 3 Assembly of the Polyaryl Carboxylic Fullerene Derivatives in Solutions in the Form of Vesicles

[0465] Method:

[0466] 1 mg of a polyaryl carboxylic fullerene derivative powdery sample was weighed and added to 1 mL of a solvent (compounds 1 to 3 each were added to DMF, and compound 4 was added to acetonitrile), and the resulting mixture was subjected to an ultrasonic treatment at 30° C. to 40° C. for 1 h to 2 h to obtain a solution in which the polyaryl carboxylic fullerene derivative powdery sample was dissolved; the solution was then added dropwise to a silicon wafer, and after the solvent was evaporated, a layer of platinum of about 10 nm was sprayed onto the surface of the sample, and then the sample was observed under a scanning electron microscope.

[0467] Results:

[0468] A Tyndall phenomenon could be observed after the polyaryl carboxylic fullerene derivatives 1 to 6 each were dissolved in the respective solvent. SEM results showed that vesicles formed by compound 1 had a particle size of 60 nm to 70 nm (as shown in FIGS. 6A-6B); vesicles formed by compound 3 had a particle size of 40 nm to 50 nm (as shown in FIGS. 7A-7B); and vesicles formed by compound 4 had a particle size of about 40 nm (as shown in FIGS. 8A-8B), indicating that such compounds each assembled in the form of vesicles in a specified solution.