Anti-coronavirus polypeptides and their uses

Abstract

The present application relates to polypeptides and uses thereof for preventing or treating novel coronavirus. [Solution] The polypeptide is at least one of P3 polypeptide and P3-1 polypeptide, P3-2 polypeptide, P3-3 polypeptide, P3-4 polypeptide, and P3-5 polypeptide induced by the P3 polypeptide, and their amino acid sequences are respectively set forth in SEQ ID NOs: 1 to 6. The polypeptide of the present application has a strong inhibitory effect against both the parent strain of the novel coronavirus and multiple mutant strains, and can therefore be used to produce drugs or vaccines for preventing and / or treating diseases caused by the novel coronavirus, and the polypeptide is expected to have the potential to prevent and / or treat new mutant strains and sarbecoviruses that will appear in the future.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to a Chinese patent application filed on August 16, 2021, bearing application number 202110939740.1 and entitled "Anti-coronavirus polypeptide and use thereof," which is incorporated herein by reference in its entirety. [Technical Field]

[0002] This application relates to the field of biopharmaceuticals, and in particular to polypeptides for preventing or treating the novel coronavirus, polynucleotides encoding the same, nucleic acid constructs comprising the polynucleotides, expression vectors comprising the nucleic acid constructs, transformed cells and pharmaceutical compositions comprising the foregoing, as well as their use in the manufacture of drugs or vaccines for preventing and / or treating the novel coronavirus. [Background technology]

[0003] The novel coronavirus pneumonia (COVID-19) pandemic poses a serious threat to global public health and human health. The SARS-CoV-2 virus is the pathogen causing this pandemic and the third coronavirus to cause a human pandemic since the beginning of the 21st century. Currently, several vaccines and monoclonal antibody drugs against SARS-CoV-2 have received emergency use authorization. However, numerous studies have shown that their protective efficacy against widely circulating SARS-CoV-2 variants, such as alpha (B.1.1.7), beta (B.1.351), and gamma (P.1), is reduced. Therefore, there is an urgent need to develop broad-spectrum drugs or vaccines against currently circulating and future emerging variants.

[0004] Currently, several coronaviruses related to the novel coronavirus (belonging to the sarbecovirus) have been reported, such as bat-derived RaTG13, RmYN02, ZC45, and ZXC21, as well as pangolin-derived GX / P2V / 2017 and GD / 1 / 2019. Research has revealed that RaTG13, GX / P2V / 2017, and GD / 1 / 2019 also have the potential to infect humans. Therefore, there is an urgent need to develop drugs or vaccines against these related coronaviruses in order to deal with future pandemics. Summary of the Invention [Problem to be solved by the invention]

[0005] The object of the present application is to provide polypeptides for preventing or treating the novel coronavirus, polynucleotides encoding the same, nucleic acid constructs containing the polynucleotides, expression vectors containing the nucleic acid constructs, transformed cells and pharmaceutical compositions containing the above, as well as their use in the manufacture of drugs or vaccines for preventing or treating the novel coronavirus.

[0006] The coronavirus spike (S) protein plays a key role in mediating the viral entry process and is divided into two subunits, S1 and S2. The S1 subunit recognizes receptors, while the S2 subunit mediates membrane fusion between the viral envelope and the host cell membrane. The heptapeptide repeats HR1 and HR2 in S2 mediate membrane fusion by forming a six-helix bundle structure. Research has shown that the addition of exogenous HR1 or HR2 polypeptides can inhibit the formation of the six-helix bundle structure by the viral HR1 and HR2, further inhibiting the membrane fusion process. The present polypeptide, designed based on the HR2 region of the SARS-CoV-2 S protein, effectively inhibits the S protein-mediated membrane fusion process and inhibits novel coronavirus infection. [Means for solving the problem]

[0007] To achieve the above objectives, the present application provides the following technical solutions: In a first aspect, the present application provides a polypeptide for anti-novel coronavirus, the polypeptide being at least one of P3 polypeptide and P3-1 polypeptide, P3-2 polypeptide, P3-3 polypeptide, P3-4 polypeptide, and P3-5 polypeptide derived therefrom, with the proviso that: The amino acid sequence of the P3 polypeptide is ISGINASVVNIQKEIDRLNEVAKNLNESLIDLQEL (set forth in SEQ ID NO: 1), The amino acid sequence of the P3-1 polypeptide is ISGINASVVNIQKEIDRLNEVAKNLNESLIDLKEL (shown in SEQ ID NO: 2), The amino acid sequence of the P3-2 polypeptide is VDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQEL (shown in SEQ ID NO: 3), The amino acid sequence of the P3-3 polypeptide is VKFGDISGINASVVNIKEEIDRLYEVVKNLNESLIDLQEL (shown in SEQ ID NO: 4), The amino acid sequence of the P3-4 polypeptide is ISGINASVVNIKEEIDRLNEVAKNLNESLIDLQEL (shown in SEQ ID NO: 5), The amino acid sequence of the P3-5 polypeptide is ISGINASVVNIQKEIDRLNEVAKELNESLIDLQEL (shown in SEQ ID NO: 6).

[0008] In a preferred specific embodiment, the polypeptide is at least one selected from a P3-1 polypeptide, a P3-2 polypeptide, a P3-3 polypeptide, and a P3-4 polypeptide; Preferably, the polypeptide is a P3-3 polypeptide.

[0009] In another preferred embodiment, the polypeptide is linked to a carrier protein, such as human serum albumin, the purpose of adding the carrier protein being to extend the half-life of the polypeptide in the body.

[0010] In a second aspect, the present application provides a polynucleotide encoding a polypeptide according to the first aspect.

[0011] Preferably, the polynucleotide is DNA or mRNA, In a specific embodiment, the polynucleotide comprises a nucleotide sequence selected from the nucleotide sequences set forth in SEQ ID NOs: 7 to 12; Specifically, the nucleotide sequence encoding the P3 polypeptide is set forth in SEQ ID NO:7, the nucleotide sequence encoding the P3-1 polypeptide is set forth in SEQ ID NO:8, the nucleotide sequence encoding the P3-2 polypeptide is set forth in SEQ ID NO:9, the nucleotide sequence encoding the P3-3 polypeptide is set forth in SEQ ID NO:10, the nucleotide sequence encoding the P3-4 polypeptide is set forth in SEQ ID NO:11, and the nucleotide sequence encoding the P3-5 polypeptide is set forth in SEQ ID NO:12. SEQ ID NO:7: ATTAGCGGCATTAACGCCTCTGTGGTGAACATTCAGAAGGAGATTGACAGACTGAACGAGGTGGCCAAGAACCTGAACGAGTCTCTCATTGACCTGCAGGAGCTG; SEQ ID NO:8: ATTAGCGGCATTAACGCCTCTGTGGTGAACATTCAGAAGGAGATTGACAGACTGAACGAGGTGGCCAAGAACCTGAACGAGTCTCTCATTGACCTGAAGGAGCTG; SEQ ID NO:9: GTGGACCTGGGCGACATTAGCGGCATTAACGCCTCTGTGGTGAACATTCAGAAGGAGATTGACAGACTGAACGAGGTGGCCAAGAACCTGAACGAGTCTCTCATTGACCTGCAGGAGCTG; SEQ ID NO:10: GTGAAGTTCGGCGACATTAGCGGCATTAACGCCTCTGTGGTGAACATTAAGGAGGAGATTGACAGACTGTACGAGGTGGTGAAGAACCTGAACGAGTCTCTCATTGACCTGCAGGAGCTG; SEQ ID NO:11: ATTAGCGGCATTAACGCCTCTGTGGTGAACATTAAGGAGGAGATTGACAGACTGAACGAGGTGGCCAAGAACCTGAACGAGTCTCTCATTGACCTGCAGGAGCTG; SEQ ID NO: 12: ATTAGCGGCATTAACGCCTCTGTGGTGAACATTCAGAAGGAGATTGACAGACTGAACGAGGTGGCCAAGGAGCTGAACGAGTCTCTCATTGACCTGCAGGAGCTG.

[0012] In a third aspect, the present application provides a nucleic acid construct comprising a polynucleotide according to the second aspect above.

[0013] Preferably, the nucleic acid construct further comprises at least one expression control element operably linked to the polynucleotide.

[0014] In a fourth aspect, the present application provides an expression vector comprising the nucleic acid construct according to the third aspect above.

[0015] In a fifth aspect, the present application provides a transformed cell comprising a polynucleotide according to the second aspect above, a nucleic acid construct according to the third aspect above, or an expression vector according to the fourth aspect above.

[0016] In a sixth aspect, the present application provides a pharmaceutical composition comprising a polypeptide according to the first aspect above, a polynucleotide according to the second aspect above, a nucleic acid construct according to the third aspect above, an expression vector according to the fourth aspect above or a transformed cell according to the fifth aspect above, together with a pharmaceutically acceptable carrier and / or excipient.

[0017] Preferably, the pharmaceutical composition is in the form of a nasal spray formulation or an oral formulation or a parenteral formulation, More preferably, the oral formulation is selected from tablets, capsules, granules, suspensions and pills; More preferably, the parenteral formulation is an injectable or bolus-administrable formulation; Preferably, the pharmaceutical composition is a vaccine composition.

[0018] In a seventh aspect, the present application provides use of a polypeptide according to the first aspect above, a polynucleotide according to the second aspect above, a nucleic acid construct according to the third aspect above, an expression vector according to the fourth aspect above, or a transformed cell according to the fifth aspect above, or a pharmaceutical composition according to the sixth aspect above, in the manufacture of a medicament or vaccine for the prevention and / or treatment of a novel coronavirus infection.

[0019] Preferably, the novel coronavirus is a SARS-CoV-2 parent strain and / or a SARS-CoV-2 mutant strain and / or a sarbecovirus; More preferably, the SARS-CoV-2 variant is the D614G strain, alpha (B.1.1.7), beta (B.1.351), gamma (P.1), kappa (B.1.617.1) and / or delta (B.1.617.2) strain, and the sarbecovirus is a virus of the genus Betacoronavirus, subgenus Sarbecovirus.

[0020] In an eighth aspect, the present application provides a method for preventing or treating a novel coronavirus, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of the polypeptide according to the first aspect above, the polynucleotide according to the second aspect above, the nucleic acid construct according to the third aspect above, the expression vector according to the fourth aspect above, or the transformed cell according to the fifth aspect above, or the pharmaceutical composition according to the sixth aspect above.

[0021] Preferably, the novel coronavirus is a SARS-CoV-2 parent strain and / or a SARS-CoV-2 mutant strain and / or a sarbecovirus; More preferably, the SARS-CoV-2 variant is the D614G strain, alpha (B.1.1.7), beta (B.1.351), gamma (P.1), kappa (B.1.617.1) and / or delta (B.1.617.2) strain, and the sarbecovirus is a virus of the genus Betacoronavirus, subgenus Sarbecovirus. [Effects of the Invention]

[0022] The inventors of the present application designed polypeptide P3 based on the HR2 region of the S protein of the SARS-CoV-2 virus and made a series of modifications to it (including the addition or substitution of one or more amino acids) to form derived peptides P3-1, P3-2, P3-3, P3-4, and P3-5. Polypeptide P3 and each derived peptide have strong inhibitory effects against the parent strain of SARS-CoV-2 and multiple mutant strains, and can be used to manufacture drugs or vaccines for preventing or treating the new coronavirus. It is expected that the polypeptides will also have the potential to prevent or treat new mutant strains and sarbecoviruses that may emerge in the future. [Brief explanation of the drawings]

[0023] One or more embodiments are illustratively illustrated in the images in the corresponding drawings, and these illustrative illustrations do not constitute limitations on the embodiments. As used herein, the word "exemplary" means "serving as an example, example, or illustration." Any embodiment described "exemplary" herein should not necessarily be construed as superior or better than other embodiments. [Figure 1] Sequence comparison of the HR1 and HR2 regions of the S protein of SARS-CoV-2 and related coronaviruses. [Figure 2]Inhibitory effects of different polypeptides of the present application on pseudoviruses of the parental strain of SARS-CoV-2 and its mutant strains. DETAILED DESCRIPTION OF THE INVENTION

[0024] In order to make the objectives, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions of the embodiments of the present application are described below clearly and completely. It goes without saying that the described embodiments are not all embodiments, but only some embodiments of the present application. All other embodiments that a person skilled in the art can obtain based on the embodiments of the present application without making any inventive work fall within the scope of protection of the present application. Unless otherwise clearly stated, throughout the specification and claims, the term "comprise" or its variations, such as "comprises" or "contains", shall be understood to include the elements or components described, but not to exclude other elements or components.

[0025] In addition, for the purpose of further explaining the present application, many specific details are provided in the specific embodiments described below. Those skilled in the art will understand that the present application can be implemented without certain specific details. In some examples, in order to enhance the subject matter of the present application, materials, elements, methods, means, etc. that are familiar to those skilled in the art are not described in detail.

[0026] The present application will be described in detail below.

[0027] In this application, a polypeptide P3 was designed based on the HR2 region of the S protein of SARS-CoV-2, and a series of modifications were made to the P3 polypeptide (including the addition or substitution of one or more amino acids), and the modified polypeptides were designated P3-1, P3-2, P3-3, P3-4, and P3-5, respectively. They were directly synthesized by methods commonly used in the art.

[0028] Experimental equipment and materials: HEK293T cells (purchased from the ATCC cell bank), Skeleton virus G*VSV-delG packaged by pseudovirus (purchased from Wuhan Privilege Neuroscience and Technology Co., Ltd.); Eukaryotic protein pCAGGS expression vector (provided by Suzhou Jin Weizhi Co., Ltd.); Hela hACE2 cells (hACE2 stable transfected cell line constructed by the applicant, see Example 1), Polypeptide P3, whose coding sequence is SEQ ID NO: 1, synthesized by Shenzhen Hanyu Pharmaceutical Co., Ltd.; Polypeptide P3-1, whose coding sequence is SEQ ID NO: 2, synthesized by Shenzhen Hanyu Pharmaceutical Co., Ltd.; Polypeptide P3-2, whose coding sequence is SEQ ID NO: 3, synthesized by Shenzhen Hanyu Pharmaceutical Co., Ltd.; Polypeptide P3-3, whose coding sequence is SEQ ID NO: 4, synthesized by Shenzhen Hanyu Pharmaceutical Co., Ltd.; Polypeptide P3-4, whose coding sequence is SEQ ID NO: 5, synthesized by Shenzhen Hanyu Pharmaceutical Co., Ltd.; Polypeptide P3-5, whose coding sequence is SEQ ID NO:6, synthesized by Shenzhen Hanyu Pharmaceutical Co., Ltd. The sequences of the above six polypeptides are shown in Table 1, in which the bolded parts are the amino acids that have been added or substituted relative to polypeptide P3. [Table 1]

[0029] The present pseudoviruses of the parental SARS-CoV-2 strain and its mutant strains were packaged in the inventor's laboratory (see Example 2).

[0030] Example 1 Construction of Hela hACE2 cell line First, the hACE2 gene was fused with a flag tag and expressed. The sequence was artificially synthesized (synthesis services were provided by Jin Weizhi, Suzhou), and then ligated into the pLVX lentiviral expression vector to obtain the hACE2 expression vector pLVX-hACE2.

[0031] The construction of the cell line was as follows. a. Cell preparation. HEK293T cells were plated onto a 6-well cell culture plate one day in advance to reach 80-90% confluence the next day.

[0032] b. Transfection: The target plasmids (pLVX-hACE2 and the lentiviral-packaged plasmids psPAX2 and pMD2.G) were mixed at a ratio of 2:2:1, and the plasmids were mixed uniformly with PEI at a ratio of 1:3. After transfection, the cell culture medium was replaced with DMEM containing 2% FBS after 4-6 hours, and the cells were cultured for another 48 hours.

[0033] c. Virus harvest and infection: The cell culture supernatant contained packaged lentivirus, which was filtered through a 0.45 μm sterile filter to remove cell debris, and added to HeLa cells prepared one day earlier.

[0034] d. Screening of clones: HeLa cells were cultured in a medium containing 2 μg / mL of puromycin to screen for positive clones.

[0035] e. Establishment of monoclonal cell lines. Monoclonal cells were selected using flow cytometry, placed in a 96-well cell culture plate, and cultured until single cell clusters were visible under a microscope. After expansion, the target gene was sequenced and other methods were used to confirm the identity of the cells.

[0036] Example 2 Packaging of pseudoviruses of SARS-CoV-2 parent and mutant strains Preparation of expression plasmid for truncated S protein 1) The nucleotides encoding the 18th amino acid from the end of the S protein of the SARS-CoV-2 parent strain (WT) and mutant strains (D614G, alpha (B.1.1.7), beta (B.1.351), gamma (P.1), kappa (B.1.617.1), and delta (B.1.617.2)) were removed to obtain the nucleotide sequences SARS-CoV-2-WT-S-del18, D614G-S-del18, B.1.1.7-S-del18, B.1.351-S-del18, P.1-S-del18, B.1.617.1-S-del18, and B.1.617.2-S-del18, respectively (these sequences are shown in SEQ ID NOs: 13 to 19, respectively), which were synthesized by Suzhou Jinweizhi Co., Ltd.

[0037] 2) Each nucleotide sequence obtained in 1) was cloned into the pCAGGS expression vector to obtain the expression plasmids pCAGGS-SARS-CoV-2-WT-S-del18, pCAGGS-D614G-S-del18, pCAGGS-B.1.1.7-S-del18, pCAGGS-B.1.351-S-del18, pCAGGS-P.1-S-del18, pCAGGS-B.1.617.1-S-del18, and pCAGGS-B.1.617.2-S-del18.

[0038] Packaging of pseudoviruses of SARS-CoV-2 parent and mutant strains a. Cell preparation: HEK293T cells were plated on a 10 cm cell culture dish to reach approximately 80% confluence the next day.

[0039] b. Transfection. The expression plasmids for each S protein obtained in step 2) above were transfected using PEI at 30 μg of plasmid per 10 cm cell culture dish. The target plasmid and PEI were uniformly mixed at a ratio of 1:3 and then transfected. The culture medium was replaced every 4 to 6 hours (DMEM medium containing 10% FBS) and the cells were cultured at 37°C for 24 hours.

[0040] c. Virus addition: The skeleton virus G*VSV-delG (purchased from Wuhan Privilege Brain Science and Technology Co., Ltd.) packaged by the pseudovirus was added to the transfected HEK293T cells and incubated at 37°C for 2 hours. The culture medium was then replaced with DMEM containing 10% FBS, and VSV-G antibody was added (hybridoma cells expressing this antibody were purchased from the ATCC Cell Bank). The cells were then cultured in an incubator for 30 hours.

[0041] d. Virus collection: The supernatant was collected and centrifuged at 3000 rpm for 10 minutes, filtered through a 0.45 μm sterile filter in a clean bench to remove cell debris, aliquoted, and stored frozen in a −80° C. refrigerator.

[0042] Through the above steps, pseudoviruses of the SARS-CoV-2 parent strain (SARS-CoV-2 WT) and mutant strains (D614G, alpha (B.1.1.7), beta (B.1.351), gamma (P.1), kappa (B.1.617.1), and delta (B.1.617.2)) were obtained, respectively.

[0043] Example 3 Evaluation of the inhibitory effect of polypeptides against pseudoviruses of the parental strain of SARS-CoV-2 and its mutant strains The purpose of this example was to determine the inhibitory effect of polypeptide P3 and its five derived polypeptides (P3-1, P3-2, P3-3, P3-4 and P3-5) against pseudoviruses of the parent strain (SARS-CoV-2 WT) and mutant strains (D614G, alpha (B.1.1.7), beta (B.1.351), gamma (P.1), kappa (B.1.617.1) and delta (B.1.617.2)).

[0044] The experimental groups were Hela hACE2 cells (blank control group, cells not infected with virus), Hela hACE2 cells + parental or mutant pseudovirus + DMEM medium (negative control group, cells infected with virus but not treated with polypeptide), Hela hACE2 cells + parental or mutant pseudovirus + polypeptide P3 (P3 treatment group), Hela hACE2 cells + parental or mutant pseudovirus + P3-1 (P3-1 treatment group), Hela hACE2 cells + parental or mutant pseudovirus + P3-2 (P3-2 treatment group), Hela hACE2 cells + parental or mutant pseudovirus + P3-3 (P3-3 treatment group), Hela hACE2 cells + parental or mutant pseudovirus + P3-4 (P3-4 treatment group), and Hela hACE2 cells + parental or mutant pseudovirus + P3-5 (P3-5 treatment group).

[0045] Preparation of polypeptide stock solutions: Polypeptides P3, P3-1, P3-2, P3-3, P3-4, and P3-5 were prepared as 10 mM stock solutions in DMSO, and the stock solutions were further diluted to 20 μM with DMEM medium containing 10% FBS, which was used as stock solutions for further gradient dilutions.

[0046] Preparation of polypeptide gradient dilutions: The 20 μM stock polypeptide solutions were diluted two-fold with DMEM medium containing 10% FBS to create nine gradients (10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.625 μM, 0.3125 μM, 0.156 μM, 0.078 μM, and 0.039 μM, respectively). Each gradient was performed in triplicate, with each well containing 50 μL.

[0047] Determination of the pseudovirus dose. The pseudovirus stock solution and a series of dilutions of the parent or mutant SARS-CoV-2 strain were quantified in HeLa hACE2 cells, and the dilution level at which 1,000 FFU appeared was used as the virus dose for evaluating the inhibitory effect of the polypeptide (dilution factors ranged from 6 to 20 times).

[0048] The virus inhibitory effect was measured as follows. a. HeLa hACE2 cells were plated in a 96-well cell culture plate and cultured to reach 80-90% confluence the next day.

[0049] b. Based on the above experimental grouping, an equal volume (50 μL) of the quantified pseudovirus dilution was added to each of the above polypeptide gradient dilutions, mixed uniformly, and then incubated at 37°C for 1 hour.

[0050] c. The supernatant was carefully discarded from the 96-well cell culture plate in step a, and the polypeptide-pseudovirus liquid mixture in step b was added (100 μL / well), followed by further culturing in an incubator for 15 to 24 hours.

[0051] d. Using a high-content microscope, the number of infected cells was counted, and the inhibition rate of each polypeptide at different concentrations was calculated. The IC value of each polypeptide was also calculated using GraphPad. 50 The results are shown in Table 2. The fold improvement in the inhibitory effect of the derived polypeptides P3-1 to P3-5 on the pseudovirus of the parent strain of SARS-CoV-2 was also calculated (see Figure 2). [Table 2] [Table 3]

[0052] As can be seen from Tables 2 and 3, the P3 polypeptide had a very good inhibitory effect on both the parent strain and multiple mutant strains of SARS-CoV-2 (0.45 μM ≦ IC 50Compared with the P3 polypeptide, the inhibitory effects of the derived polypeptides P3-1, P3-2, P3-3, and P3-4 against the parent strain and multiple mutant strains of SARS-CoV-2 were all improved to varying degrees, with the most obvious improvement being seen for polypeptide P3-3. The inhibitory effect of P3-5 was slightly reduced, but its inhibitory effect against kappa mutant strains was slightly greater than that of P3.

[0053] Tables 2 and 3 show that the polypeptides of the present application can be used to prevent or treat diseases caused by the novel coronavirus, and therefore it is presumed that they can also be widely used for diseases caused by sarbecoviruses.

[0054] During the process of SARS-CoV-2 invading host cells, the polypeptide of the present application inhibits viral entry by inhibiting the membrane fusion process mediated by the S protein. It is suitable for the treatment or prevention of the parent strain and mutant strains of the new coronavirus, such as D614G, B.1.1.7 (alpha), B.1.351 (beta), P.1 (gamma), kappa (B.1.617.1) or delta (B.1.617.2), and is also effective in inhibiting sarbecoviruses and new new coronavirus mutant strains that may emerge in the future, and is expected to have wide potential useful value.

[0055] Finally, it should be clearly stated that the above embodiments are only intended to illustrate, not to limit, the technical solutions of the present application. Although the above embodiments have described the present application in detail, those skilled in the art will understand that the technical solutions described in each embodiment may be modified or some technical features may be replaced with equivalents. Such modifications or replacements will not deviate from the spirit and scope of the corresponding technical solutions of each embodiment of the present application. [Industrial Applicability]

[0056] The present application provides polypeptides for preventing or treating novel coronavirus and uses thereof, the polypeptides being P3 polypeptides and at least one of P3-1 polypeptide, P3-2 polypeptide, P3-3 polypeptide, P3-4 polypeptide, and P3-5 polypeptide derived from P3 polypeptide. The polypeptides provided by the present application have strong inhibitory effects against both the parent strain of the novel coronavirus and multiple mutant strains, and can therefore be used to produce drugs or vaccines for preventing and / or treating diseases caused by novel coronaviruses. These drugs are also expected to have the potential to prevent and / or treat new mutant strains and sarbecoviruses that may emerge in the future, making them highly promising and valuable for clinical use.

Claims

1. An anti-coronavirus polypeptide, at least one of a P3-2 polypeptide, a P3-1 polypeptide, a P3-3 polypeptide, and a P3-4 polypeptide; The amino acid sequence of the P3-2 polypeptide is set forth in SEQ ID NO:3, The amino acid sequence of the P3-1 polypeptide is shown in SEQ ID NO:2, The amino acid sequence of the P3-3 polypeptide is set forth in SEQ ID NO:4, The amino acid sequence of the P3-4 polypeptide is set forth in SEQ ID NO:

5. Polypeptide.

2. The polypeptide of claim 1, wherein the polypeptide is linked to a carrier protein.

3. The polypeptide described in Claim 2, characterized in that the carrier protein is human serum albumin.

4. A polynucleotide encoding the polypeptide according to any one of claims 1 to 3.

5. The polynucleotide of claim 4, wherein the polynucleotide is DNA or mRNA.

6. The polynucleotide described in claim 4, wherein the polynucleotide comprises a nucleotide sequence selected from the nucleotide sequences shown in SEQ ID NOs: 8 to 11.

7. A nucleic acid construct comprising the polynucleotide according to any one of claims 4 to 6.

8. 8. The nucleic acid construct of claim 7, further comprising at least one expression control element operably linked to the polynucleotide.

9. An expression vector comprising the nucleic acid construct of claim 7 or 8.

10. A transformed cell comprising the polynucleotide according to any one of claims 4 to 6, the nucleic acid construct according to claim 7 or 8, or the expression vector according to claim 9.

11. A pharmaceutical composition comprising the polypeptide according to any one of claims 1 to 3, the polynucleotide according to any one of claims 4 to 6, the nucleic acid construct according to claim 7 or 8, the expression vector according to claim 9, or the transformed cell according to claim 10, and a pharmaceutically acceptable carrier and / or excipient.

12. 12. The pharmaceutical composition according to claim 11, wherein the pharmaceutical composition is in the form of a nasal spray formulation, an oral formulation, or a parenteral formulation.

13. The oral formulation is selected from tablets, capsules, granules, suspensions, and pills, 13. The pharmaceutical composition of claim 12, wherein the parenteral formulation is an injectable or bolus-administerable formulation.

14. The pharmaceutical composition of claim 11, wherein the pharmaceutical composition is a vaccine composition.

15. Use of a polypeptide according to any one of claims 1 to 3, a polynucleotide according to any one of claims 4 to 6, a nucleic acid construct according to claim 7 or 8, an expression vector according to claim 9, a transformed cell according to claim 10 or a pharmaceutical composition according to any one of claims 11 to 14 in the manufacture of a drug or vaccine for preventing and / or treating a novel coronavirus.

16. The use according to claim 15, characterized in that the novel coronavirus is a SARS-CoV-2 parent strain and / or a SARS-CoV-2 mutant strain and / or a sarbecovirus.

17. The use according to claim 16, wherein the SARS-CoV-2 mutant strain is a D614G strain, an alpha (B.1.1.7), a beta (B.1.351), a gamma (P.1), a kappa (B.1.617.1) and / or a delta (B.1.617.2) strain, and the sarbecovirus is a virus of the genus Betacoronavirus, subgenus Sarbecovirus.

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