Application of sulbactam in preparation of medicine for treating novel coronavirus infection

Abstract

The invention discloses an application of sulbactam in preparation of a medicine for treating novel coronavirus infection, which is characterized in that the expression or activity of USP22 is inhibited, and the deubiquitination level of N protein is reduced, so that the stability of the N protein is reduced, and the aim of inhibiting virus survival and transmission is fulfilled; according to the invention, the technical blank of the existing treatment means in the aspect of N protein specific targeting regulation is filled, and a new thought is provided for anti-new coronavirus therapy.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to the use of sulbactam in the preparation of drugs for treating novel coronavirus infection. Background Art

[0002] Current studies have shown that the nucleocapsid protein (N protein) of SARS-CoV-2 plays an important role in the life cycle of the virus, including viral RNA packaging, replication, genome assembly, and evasion of host immune responses. However, in existing antiviral treatments, the mechanism of action of the N protein has not been fully utilized, especially the regulatory mechanism of its ubiquitination and deubiquitination process is still significantly insufficient. Deubiquitination of the nucleocapsid protein contributes to its stability in host cells, which directly affects the survival and spread of the virus.

[0003] In addition, the N protein can evade the host's defense mechanisms by inhibiting the host's immune signals (such as interferon responses). Studies have shown that the SARS-CoV-2 N protein can inhibit PKR-mediated cellular stress responses and interfere with the formation of stress granules (SGs), thereby limiting the activation of host immune sensing and antiviral responses. Although some current antiviral drugs can inhibit viral replication, due to this immune escape mechanism of the N protein, the targeting effect of these drugs on the N protein is often ineffective, especially when dealing with mutant strains.

[0004] Therefore, the existing technology lacks effective therapeutic strategies for regulating the deubiquitination of N protein and enhancing the host immune response. By inhibiting the deubiquitination of N protein to reduce its stability (such as through the action of USP22 deubiquitinase), it can effectively interfere with the stability and reproduction of the virus, thereby improving the antiviral effect. This idea provides an important research direction for the design of new antiviral therapies and fills the gap in the regulatory mechanism of existing antiviral drugs against N protein.

[0005] In addition, studies have shown that sulbactam can play an important role in treating secondary bacterial infections in patients with severe COVID-19, especially for patients with resistant bacterial infections such as Acinetobacter baumannii. Recent multi-drug combination studies have found that sulbactam combined with other antibiotics (such as polymyxins and carbapenems) can help reduce the mortality rate of secondary infections in critically ill patients. This application is mainly aimed at severe bacterial infections caused by COVID-19, rather than direct antiviral treatment.

[0006] However, to date, there have been no studies on the antiviral effects of sulbactam. Summary of the invention

[0007] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0008] In view of the above problems and / or the problems existing in the prior art, the present invention is proposed.

[0009] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide the use of sulbactam in the preparation of drugs for treating novel coronavirus infection.

[0010] As a preferred embodiment of the application described in the present invention, the disease caused by the new coronavirus is a respiratory tract infection.

[0011] As a preferred embodiment of the application of the present invention, the application concentration of sulbactam at the cellular level is 4 mg / ml.

[0012] As a preferred embodiment of the application of the present invention, the deubiquitinating enzyme USP22 interacts with the N protein on the surface of the virus to reduce the ubiquitination level of the N protein.

[0013] As a preferred embodiment of the application of the present invention, the deubiquitinase USP22 interacts with the N protein on the surface of the virus to prevent the degradation of the N protein.

[0014] As a preferred embodiment of the application of the present invention, the sulbactam inhibits the deubiquitination function of USP22, interferes with the stability of the N protein and inhibits viral replication.

[0015] As a preferred embodiment of the application of the present invention, the sulbactam downregulates the expression levels of USP22 and N proteins in cells.

[0016] As a preferred embodiment of the application of the present invention, the effect of sulbactam on intracellular USP22 and N protein is dose-dependent.

[0017] As a preferred embodiment of the application of the present invention, the sulbactam downregulates the USP22 protein level, thereby reducing the N protein level.

[0018] As a preferred embodiment of the application of the present invention, the application of sulbactam in inhibiting the expression of deubiquitinating enzyme USP22 in the preparation of therapeutic antiviral drugs.

[0019] Another aspect of the present invention overcomes the deficiencies in the prior art and provides a composition of sulbactam and an antiviral drug for use as an active ingredient in the preparation of a drug for treating diseases caused by the new coronavirus.

[0020] As a preferred embodiment of the application of the present invention, the antiviral drug is selected from one or more of ceftizoxime, ceftazidime, ceftriaxone, penicillin, latamoxef, meropenem, ceftriaxone / sulbactam, piperacillin, and cefotaxime.

[0021] Beneficial effects of the present invention:

[0022] 1. This invention reveals for the first time the mechanism of action of SARS-CoV-2 nucleocapsid protein (N protein) that depends on the host deubiquitinase USP22, indicating that the stability of N protein can be reduced by regulating USP22 expression. Most existing antiviral drugs focus on viral replication or protein synthesis, while this patent provides a new antiviral target through the inhibitory effect of USP22, thereby interfering with the stability of N protein and inhibiting viral replication;

[0023] 2. The present invention utilizes the USP22 inhibitory ability of sulbactam to show new application potential in the treatment of new coronavirus, and can achieve good antiviral effects without being used in combination with other antibiotics;

[0024] 3. Compared with newly developed antiviral drugs, the sulbactam of the present invention is easier to promote and use in clinical practice, thus accelerating its practical application and industrialization process;

[0025] 4. Sulbactam's inhibition of USP22 shows significant dose-dependency, and the dose can be flexibly adjusted to achieve the best antiviral effect. This dose-dependency feature facilitates precise use in patients with different degrees of infection and reduces the risk of side effects;

[0026] 5. The technical solution disclosed in this patent is suitable for combating SARS-CoV-2, and has an inhibitory effect on viral stability and replication ability. For strains known to have immune escape characteristics, sulbactam targets the host protein USP22 to reduce the stability of the N protein, which is expected to overcome the limitations of existing therapies and provide wider applicability for the new coronavirus and its variants. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor. Among them:

[0028] Figure 1 The N protein plasmid and the deubiquitinating enzyme plasmid in the DUB library were transfected into HEK-293T cells in Example 1 of the present invention, and the N protein level was detected by Western Blot technology.

[0029] Figure 2 The N protein plasmid and the USP22 plasmid were transfected into HEK-293T cells in Example 1 of the present invention, and the interaction between the N protein and USP22 was detected by immunoprecipitation technology.

[0030] Figure 3 The N protein level was detected by Western Blot technique using the N protein plasmids and increasing concentrations of USP22 plasmids in HEK-293T and A549 cells in Example 1 of the present invention.

[0031] Figure 4 In the HEK-293T and A549 cells lacking Usp22 in Example 1 of the present invention, N protein was transfected and the level of N protein was detected by Western Blot technology.

[0032] Figure 5 The ubiquitination level of N protein was detected by immunoprecipitation technique using N protein plasmids and increasing concentrations of USP22 plasmids and Ub plasmids exogenously transferred into HEK-293T cells in Example 1 of the present invention.

[0033] Figure 6 The N protein plasmid was transfected into HEK-293T cells lacking Usp22 in Example 1 of the present invention, and the ubiquitination level of the N protein was detected by immunoprecipitation technology.

[0034] Figure 7 In Example 2 of the present invention, HEK-293T cells were treated with 8 antibiotics, and the USP22 protein level was detected using Western Blot technology.

[0035] Figure 8 The HEK-293T cells in Example 2 of the present invention were treated with increasing concentrations of cefotaxime sulbactam, and the levels of USP22 and N proteins were detected using Western Blot technology.

[0036] Fig. 9 The HEK-293T cells in Example 2 of the present invention were treated with increasing concentrations of cefotaxime, and the USP22 protein level was detected using Western Blot technology.

[0037] Fig.10 HEK-293T and A549 cells in Example 2 of the present invention were treated with increasing concentrations of sulbactam, and the USP22 protein level was detected using Western Blot technology.

[0038] Fig.11 HEK-293T cells in Example 2 of the present invention were treated with the same concentration of sulbactam for different time periods, and the USP22 protein level was detected using Western Blot technology.

[0039] Fig.12 HEK-293T and A549 cells were transfected with N protein plasmid in Example 3 of the present invention, and then treated with increasing concentrations of sulbactam. The levels of N protein and USP22 protein were detected by Western Blot technology.

[0040] Fig.13 After the mice in Example 4 of the present invention were injected with sulbactam or normal saline, the USP22 protein level in the lung tissue of the mice was detected by Western Blot technology.

[0041] Fig.14 The mice were infected by intranasal drops with adenovirus expressing N protein after modification in Example 4 of the present invention, and then injected with the same dose of sulbactam, while the control group was injected with normal saline. The levels of N protein and USP22 protein in the lung tissue of mice were detected by Western Blot technology. DETAILED DESCRIPTION

[0042] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with specific embodiments.

[0043] In the following description, many specific details are set forth to facilitate a full understanding of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. The present invention can also be implemented in other ways different from those described herein, and those skilled in the art can make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed below.

[0044] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0045] Raw materials and reagents used in the present invention:

[0046] Drugs: Cefuroxime (Sinopharm Zhijun Pharmaceutical Co., Ltd.); ceftazidime (Tiger Pharmaceuticals Industrial Development Co., Ltd.); ceftriaxone (Haikou Qili Pharmaceutical Co., Ltd.); penicillin (Shijiazhuang Pharmaceutical Group Zhongnuo Pharmaceutical Co., Ltd.); latamoxef (Taiji Southwest Co., Ltd.); meropenem (Shenzhen Haibin Pharmaceutical Co., Ltd.); ceftriaxone (Xiangbei Wellman Pharmaceutical Co., Ltd.); piperacillin (Suzhou Erye Pharmaceutical Co., Ltd.); ceftriaxone (Yuekang Pharmaceutical Group Co., Ltd.); sulbactam (Chengdu Beite Pharmaceutical Co., Ltd.).

[0047] Reagents: NEM (Thermo Scientific; Cat#23030); RIPA (Biyuntian Biotechnology; Cat#P0013K); Longtrans liposome transfection reagent (Wuxi Yaokemei Biotechnology Co., Ltd.); Protein G beads (Millipore; Cat#16-266).

[0048] Virus: Adenovirus-2019-nCoV-N (Vigen Biotech Co., Ltd.).

[0049] Antibodies: anti-USP22 (Abcam, Cat#ab195289), anti-HA (Abcam, Cat#ab9110), anti-Flag (Sigma, Cat#F7425), anti-Myc (Abmart, Cat#M20002H), anti-GAPDH (Proteintech, Cat#60004-1-lg), anti-Ubiquitin (Santa Cruz, Cat#sc-66180), anti-SARS-CoV-2NP (ABclonal, Cat#A20142).

[0050] Experiments involved in the present invention:

[0051] 1. Experimental Animals

[0052] This animal experiment was authorized by the Ethics Committee of the First Affiliated Hospital of Soochow University and complied with the ARRIVE guidelines. C57BL6 / J adult male mice weighed 20-25 g. Animals were housed under standard animal conditions with a temperature of 25 ± 1 °C, a humidity of 50-60%, a light / dark cycle of 12 h, and free access to food and water. All animals were randomly assigned to different groups, and the data were evaluated under blind conditions.

[0053] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in the field of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields.

[0054] Example 1

[0055] 1. Screening for deubiquitinases that regulate N protein

[0056] Using the DUB library, potential DUBs of N protein (amino acid sequence as shown in SEQ ID NO.1) were preliminarily screened, that is, Flag-NP plasmid and DUB plasmid in the library were co-transfected into HEK293T cells.

[0057] HEK293T cells were plated on 12-well cell culture plates (about 0.5×10 6 / well), transfected with LongTrans (Ucallm); lysed the cells with RIPA lysis buffer after 24 hours, extracted total protein; lysed the cells with NP-40 lysis buffer, prepared protein samples, electrophoresed with SDS-PAGE, transferred to PVDF membrane, blocked with 5% skim milk powder at room temperature for 1 hour, added primary antibody and incubated at 4°C overnight, washed with PBST, added secondary antibody and incubated at room temperature for 1 hour, washed with PBST, added luminescent substrate for exposure, development and fixation, Western blot detected the expression level of N protein, used anti-Flag tag antibody and GAPDH (amino acid sequence as shown in SEQ ID NO.2) as internal reference, the results are shown in Figure 1 shown.

[0058] 2.USP22 interacts with N protein.

[0059] HEK293T cells were transfected with co-transfected Myc-NP and FH-USP22 plasmids; 24 h later, the transfected cells were lysed using RIPA lysis buffer containing protease inhibitors and 1 mM N-ethylmaleimide (NEM) to prevent protein degradation and ubiquitination removal; incubated on ice for 30 min, with occasional gentle shaking to ensure adequate lysis; then, centrifuged at 12000 rpm for 15 min to remove cell debris and collect the supernatant (total protein); 200-500 μg of total protein was taken, the corresponding antibody was added, and incubated with gentle shaking for 4 h to overnight to ensure that the antibody was fully bound to the target protein; 30-50 μL of pre-washed Protein G-agarose beads were added, and the incubation continued with gentle shaking for 2-4 h to capture the antigen-antibody complex; the precipitated beads were washed 4-5 times with cold RIPA lysis buffer, each centrifuged at 3000 rpm for 3 min to remove unbound nonspecific proteins; Western blot was performed. Blot analysis showed that the presence of N protein could be detected in samples immunoprecipitated with anti-FH-USP22 antibody, and the presence of USP22 (amino acid sequence as shown in SEQ ID NO.3) could also be detected in samples immunoprecipitated with anti-Myc-NP antibody, indicating that the two interacted with each other. Figure 2 shown.

[0060] 3. USP22 reduces the ubiquitination level of N protein, thereby preventing the degradation of N protein.

[0061] Flag-NP and different doses of FH-USP22 plasmids were overexpressed in HEK293T cells and A549 cells, respectively. The Flag antibody was used to detect the effect of FH-USP22 on the level of N protein. The results are shown in Figure 3 As shown, FH-USP22 significantly increased the level of N protein in different cell lines in a dose-dependent manner.

[0062] Usp22 - / - HEK293T and Usp22 - / - A549 cells were plated in 12-well cell culture plates (approximately 0.5 × 10 6 / well, transfected with the same dose of Flag-NP, and 24h later, the N protein level was detected by Western blot. The results were as follows Figure 4 shown.

[0063] The ubiquitination level of N protein in USP22-overexpressing and USP22-deficient cells was detected by immunoprecipitation. Figure 5 and Figure 6 shown.

[0064] In summary Figures 1 to 6 As described in the results, USP22 interacts with N protein, reduces its ubiquitination level, and thus prevents the degradation of N protein.

[0065] Example 2

[0066] 1. Eight antibiotics commonly used in clinical practice were screened to observe their effects on intracellular USP22 protein levels.

[0067] Antibiotics (ceftizoxime, ceftazidime, ceftriaxone, penicillin, latamoxef, meropenem, cefotaxime sulbactam, piperacillin) dissolved in saline at a concentration of 2 mg / ml were added to HEK293T cells, and whole cell lysates were collected after 12 hours of treatment (this antibiotic concentration is much lower than the clinical intravenous injection dose). Figure 7 As shown, ceftriaxone-sulbactam could significantly inhibit the level of intracellular USP22 protein.

[0068] Cells transfected with N protein were treated with different concentrations of cefotaxime sulbactam, and cells were collected and protein was extracted after 12 hours of treatment. The results of Western Blot detection are shown in Figure 8 As shown, with the increase of cefotaxime-sulbactam concentration, the intracellular USP22 protein level gradually decreased, and the N protein level also decreased significantly.

[0069] 2. To explore whether the inhibitory effect of ceftriaxone and sulbactam comes from the combination of the two, or whether each component alone is effective.

[0070] Cells were treated with different concentrations of cefotaxime and intracellular proteins were extracted 12 hours later. Fig. 9 As shown, cefotaxime alone failed to significantly downregulate the content of USP22.

[0071] Sulbactam was dissolved in physiological saline and treated with different concentrations of HEK293T and A549 cells, respectively. After 12 hours, the cells were lysed using RIPA lysis buffer containing protease inhibitors and 1mM N-ethylmaleimide (NEM) to prevent protein degradation and deubiquitination reactions; incubated on ice for 30 minutes, with occasional gentle shaking, centrifuged at 12000rpm for 15 minutes, cell debris was removed, and the supernatant was collected as a total protein extract; the extracted total protein was separated by SDS-PAGE electrophoresis, and then the protein was transferred to a PVDF membrane, and Western blot analysis was performed using USP22-specific antibodies and GAPDH antibodies. GAPDH was used as an internal reference protein to correct for differences in protein loading and transfer efficiency. Western blot results are shown in Figure 2. Fig.10 As shown in the figure, the expression level of USP22 protein in the sulbactam-treated group was significantly reduced, and with the increase of sulbactam concentration, the expression level of USP22 further decreased.

[0072] Sulbactam was dissolved in physiological saline and treated with cells at a concentration of 4 mg / ml for different time periods (0, 12, 24). The cells were harvested and proteins were extracted. The results of Western Blot detection are shown in Figure 11. It was found that as time went on, the USP22 protein level in cells treated with sulbactam sodium further decreased.

[0073] In summary, the combined use of ceftriaxone sodium and sulbactam did not significantly improve the downregulation effect of USP22 expression, which was equivalent to the effect of sulbactam alone. This result shows that sulbactam alone can achieve the expected inhibitory effect, while the combined use of ceftriaxone sodium did not significantly enhance the inhibitory effect on the new coronavirus.

[0074] Example 3

[0075] HEK293T cells and A549 cells were transfected with Flag-NP and treated with different concentrations of sulbactam (0, 4, 8 mg / ml) for 12 h. The intracellular proteins were extracted and the levels of N protein and USP22 protein in the cells were detected by Western Blot using GAPDH as an internal reference.

[0076] The results are as follows Fig.12 As shown, sulbactam can effectively downregulate the expression levels of USP22 and N protein in cells in a dose-dependent manner. This further proves that sulbactam inhibits the replication of the new coronavirus by inhibiting the deubiquitination function of USP22 and reducing the stability of N protein. This result supports the application prospect of sulbactam as a potential therapeutic drug for new coronavirus infection.

[0077] Example 4 Behavioral Experiment

[0078] The C57BL / 6 wild-type mice used in this example were purchased from the Animal Center of Soochow University and raised in the SPF environment of Soochow University.

[0079] Wild-type male C57BL / 6 mice (6-8 weeks) were randomly divided into two groups, 3 mice in each group. 100 mg / kg of sulbactam or the same volume of saline was injected intraperitoneally. The injection was given once a day for three days. Three days later, the lung tissue was immediately removed after the mice were euthanized and quickly rinsed several times with pre-cooled PBS (phosphate buffered saline) to remove blood and other impurities. The excess PBS was gently blotted with filter paper and an appropriate amount of lung tissue (usually 10-50 mg) was weighed. The lung tissue was placed in a 1.5 mL sterile centrifuge tube and 5-10 times the volume of ice-cold RIPA lysis buffer (usually 10 μL lysis buffer was added for 1 mg of tissue). Make sure that the lysis buffer contains protease inhibitors and NEM to prevent protein degradation and deubiquitination reactions. Use a tissue homogenizer to homogenize the lung tissue on ice until the tissue is completely broken and a uniform suspension is obtained. The homogenized sample was incubated on ice for 30 minutes, and gently vortexed every 5 minutes to ensure complete cell lysis and sufficient protein release. Place the homogenate in a precooled microcentrifuge and centrifuge at 12000rpm (or 14000x g) for 15 minutes (4°C) to remove cell debris and insoluble matter. Carefully transfer the supernatant (i.e., total protein solution) to a new 1.5mL centrifuge tube to avoid aspirating the precipitate. Divide the extracted total protein solution into multiple small tubes to avoid repeated freezing and thawing, and immediately freeze at -80°C for later use. Western Blot was used to detect the USP22 protein level in mouse lung tissue. The results are as follows: Fig.13 As shown, the USP22 protein level in the lung tissue of mice treated with sulbactam was greatly reduced.

[0080] Wild-type male C57BL / 6 mice (6-8 weeks) were randomly divided into two groups, with 5 mice in each group. After intraperitoneal injection of sodium pentobarbital, the mice were anesthetized and injected with adenovirus (1x10 11 PFU per gram body) were intranasally infected with mice. Four days later, the two groups of mice were intraperitoneally injected with 100 mg / kg of sulbactam or the same volume of saline. The injection was given once a day for three days. Three days later, the mouse lung tissue protein was extracted, and the levels of USP22 protein and N protein in the mouse lung tissue were detected by Western Blot. The results showed that the USP22 protein level in the lung tissue of mice treated with sulbactam was reduced, and the N protein level was also greatly reduced. The results are as follows Fig.14 shown.

[0081] The results of this example show that sulbactam sodium can significantly downregulate the USP22 protein level in mice, thereby reducing the SARS-CoV-2 N protein level. This effect is dose-dependent, indicating that sulbactam sodium reduces the stability of the N protein and the replication ability of the virus by inhibiting the USP22 protein.

[0082] In summary, the present invention reveals the mechanism of action of SARS-CoV-2 nucleocapsid protein (N protein) dependent on host deubiquitinase USP22, indicating that the stability of N protein can be reduced by regulating USP22 expression; most existing antiviral drugs focus on viral replication or protein synthesis, while this patent provides a new antiviral target through the inhibition of USP22, thereby interfering with the stability of N protein and inhibiting viral replication; and the present invention can achieve good antiviral effects without being used in combination with other antibiotics; in addition, sulbactam shows significant dose dependence in the inhibition of USP22, and the dose can be flexibly adjusted to achieve the best antiviral effect. This dose-dependent characteristic facilitates precise use in patients with different degrees of infection and reduces the risk of side effects; finally, the present invention has an inhibitory effect on viral stability and replication ability. For strains known to have immune escape characteristics, sulbactam targets the host protein USP22 to reduce the stability of N protein, which is expected to overcome the limitations of existing therapies and provide wider applicability for the new coronavirus and its variants.

[0083] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the present invention.

[0084] synthetic construct

[0085] Sequence Listing

[0086] SEQ ID NO.1

[0087] Artificial sequence

[0088] MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLPNNTAS

[0089] WFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMK

[0090] DLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPAN

[0091] NAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTS

[0092] PARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQGQTVTKKSAAEAS

[0093] KKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKHWPQIA

[0094] QFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHI

[0095] DAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQL

[0096] QQSMSSADSTQA

[0097] SEQ ID NO.2

[0098] MGKVKVGVNGFGRIGRLVTRAAFNSGKVDIVAINDPFIDLNYMVYMFQYD

[0099] STHGKFHGTVKAENGKLVINGNPITIFQERDPSKIKWGDAGAEYVVESTGVF

[0100] TTMEKAGAHLQGGAKRVIISAPSADAPMFVMGVNHEKYDNSLKIISNASCT

[0101] TNCLAPLAKVIHDNFGIVEGLMTTVHAITATQKTVDGPSGKLWRDGRGALQ

[0102] NIIPASTGAAKAVGKVIPELNGKLTGMAFRVPTANVSVVDLTCRLEKPAKYD

[0103] DIKKVVKQASEGPLKGILGYTEHQVVSSDFNSDTHSSTFDAGAGIALNDHF

[0104] VKLISWYDNEFGYSNRVVDLMAHMASKE

[0105] SEQ ID NO.3

[0106] MVSRPEPEGEAMDAELAVAPPGCSHLGSFKVDNWKQNLRAIYQCFVWSGT

[0107] AEARKRKAKSCICHVCGVHLNRLHSCLYCVFFGCFTKKHIHEHAKAKRHN

[0108] LAIDLMYGGIYCFLCQDYIYDKDMEIIAKEEQRKAWKMQGVGEKFSTWEP

[0109] TKRELELLKHNPKRRKITSNCTIGLRGLINLGNTCFMNCIVQALTHTPLLRDF

[0110] FLSDRHRCEMQSPSSCLVCEMSSLFQEFYSGHRSPHIPYKLLHLVWTHARHL

[0111] AGYEQQDAHEFLIAALDVLHRHCKGDDNGKKANNPNHCNCIIDQIFTGGL

[0112] QSDVTCQVCHGVSTTIDPFWDISLDLPGSSTPFWPLSPGSEGNVVNGESHVS

[0113] GTTTLTDCLRRFTRPEHLGSSAKIKCSGCHSYQESTKQLTMKKLPIVACFHL

[0114] KRFEHSAKLRRKITTYVSFPLELDMTPFMASSKESRMNGQYQQPTDSLNND

[0115] NKYSLFAVVNHQGTLESGHYTSFIRQHKDQWFKCDDAIITKASIKDVLDSEG

[0116] YLLFYHKQFLEYE。

Claims

1. The use of sulbactam in the preparation of a drug for treating novel coronavirus infection, characterized in that: The new coronavirus is the new coronavirus SARS-CoV-2.

2. The use according to claim 1, characterized in that: The disease caused by the new coronavirus is a respiratory infection.

3. The use according to claim 1, characterized in that The applied concentration of sulbactam at the cellular level is 4 mg / ml.

4. Use of a combination of sulbactam and antiviral drugs as an active ingredient in the preparation of drugs for treating diseases caused by the new coronavirus.

5. The use according to claim 4, characterized in that :The disease caused by the new coronavirus is an infectious disease or its complications caused by SARS-Cov-2.

6. The use according to claim 4, characterized in that: The antiviral drug is selected from one or more of ceftizoxime, ceftazidime, ceftriaxone, penicillin, latamoxef, meropenem, ceftriaxone / sulbactam, piperacillin, and cefotaxime.

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