CN113425724A

CN113425724A - Application of loratadine or ebastine in preparation of drugs for preventing and/or treating COVID-19 inflammation

Info

Publication number: CN113425724A
Application number: CN202110887654.0A
Authority: CN
Inventors: 王建华; 郑永唐; 吴梦丽; 刘丰亮; 李欣
Original assignee: Guangzhou Institute of Biomedicine and Health of CAS
Current assignee: Guangzhou Institute of Biomedicine and Health of CAS
Priority date: 2021-08-03
Filing date: 2021-08-03
Publication date: 2021-09-24

Abstract

The invention provides an application of loratadine or ebastine in preparing a medicament for preventing and/or treating COVID-19 inflammation. The invention is proved by in vivo and in vitro experiments that: the loratadine or ibastine can reduce inflammatory reaction caused by SARS-CoV-2spike protein by inhibiting mast cell degranulation, inhibit inflammatory factor storm, and has certain protection to tissue injury. Therefore, the loratadine or ibastine combined antiviral drug has important clinical application value in the aspect of treating severe infection of COVID-19.

Description

Application of loratadine or ebastine in preparation of drugs for preventing and/or treating COVID-19 inflammation

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of loratadine or ebastine in preparation of a medicine for preventing and/or treating COVID-19 inflammation.

Background

Loratadine (Loratadine, C)₂₂H₂₃CIN₂O₂) The system name is 4- (8-chloro-5, 6-dihydro-11H-benzo [5,6 ]]-cyclohepta [1,2-b ]]Pyridine-11-alkenyl) -1-piperidine carboxylic acid ethyl ester, the structural formula of which is shown in formula I; ibastine (Ebastine, C)₃₂H₃₉NO₂) The systematic name is 1- (4-tert-butylphenyl) -4- [4- (diphenylmethoxy) -1-piperidyl]The structural formula of the-1-butanone is shown as a formula II.

Loratadine and ibastine belong to the second generation of antihistamines and are clinically used for treating allergic diseases, including allergic rhinitis in children, rhinitis in adults for the last year, seasonal rhinitis, hay fever, chronic urticaria and the like.

The novel coronavirus (SARS-CoV-2) causes a novel coronavirus pneumonia (COVID-19), and severe people can rapidly progress to acute respiratory distress syndrome and multiple organ failure and death. The existing research results show that abnormal increase of inflammatory factors caused by virus infection and massive infiltration of inflammatory cells (also called as 'cytokine storm') in lung are important factors for organ damage and severe or death.

At present, the cytological mechanism of SARS-CoV-2 virus for inducing body inflammatory cytokine storm is not clear, and the clinical immunotherapy of COVID-19 inflammation lacks effective immunoregulation means.

Therefore, providing an effective immunomodulator to inhibit the production of inflammatory factors is of great significance for the clinical treatment of COVID-19.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide the application of loratadine or ebastine in preparing the medicament for preventing and/or treating COVID-19 inflammation. The invention verifies that the loratadine or the ebastine can inhibit the mast cell degranulation process induced by SARS-CoV-2spike-RBD protein on a plurality of levels through in vivo and in vitro experiments, thereby inhibiting the generation of inflammatory factors.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the invention provides an application of loratadine or ebastine in preparing a medicament for preventing and/or treating COVID-19 inflammation.

In-vivo and in-vitro experiments prove that both loratadine and ibastine can inhibit the generation of inflammatory factors caused by SARS-CoV-2, such as IL-6, IL-8, TNF alpha, IL-1 beta, INF gamma, CCL20, CCL5, CRP (C reactive protein) and the like, protect the lung and prevent injury. The occurrence of inflammatory factors can cause phenomena of alveolar space thickening, bleeding, mucosa abscission, transparent membrane formation, inflammatory cell infiltration and fiber exudation, and the like, and the loratadine or the ibastine can inhibit the degranulation of mast cells induced by SARS-CoV-2spike-RBD protein, thereby inhibiting the generation of the inflammatory factors.

Therefore, compared with other medicines, the loratadine or ibastine can be used as an immunomodulator, especially a mast cell degranulation inhibitor, reduces the COVID-19 inflammatory factor response, can be combined with an antiviral medicine to treat severe infection of the new coronavirus, and has important application value in treating the COVID-19.

As a preferable technical scheme of the invention, the dosage of the loratadine is set according to the actual condition of a patient, and the dosage of the loratadine is preferably 0.1-10 mg/kg/d, for example, 0.1mg/kg/d, 0.2mg/kg/d, 0.5mg/kg/d, 1mg/kg/d, 2mg/kg/d, 3mg/kg/d, 5mg/kg/d, 6mg/kg/d, 7mg/kg/d, 8mg/kg/d or 10mg/kg/d, etc., preferably 0.2-0.5 mg/kg/d.

In the mouse experiment, the concentration is 5-20 mg/kg/d, for example, 5mg/kg/d, 6mg/kg/d, 8mg/kg/d, 10mg/kg/d, 12mg/kg/d, 14mg/kg/d, 15mg/kg/d, 16mg/kg/d, 18mg/kg/d or 20 mg/kg/d.

As a preferable embodiment of the present invention, the dose of ebastine is set according to the actual condition of the patient, and the dose of ebastine is preferably 0.1 to 20mg/kg/d, and may be, for example, 0.1mg/kg/d, 0.2mg/kg/d, 0.5mg/kg/d, 1mg/kg/d, 2mg/kg/d, 3mg/kg/d, 5mg/kg/d, 6mg/kg/d, 8mg/kg/d, 10mg/kg/d, 12mg/kg/d, 15mg/kg/d, 18mg/kg/d or 20mg/kg/d, and preferably 0.1 to 1 mg/kg/d.

In the mouse experiment process, the dose of the ebastine is 2-6 mg/kg/d, and can be, for example, 2mg/kg/d, 2.5mg/kg/d, 3mg/kg/d, 3.5mg/kg/d, 4mg/kg/d, 4.5mg/kg/d, 5mg/kg/d, 5.5mg/kg/d or6 mg/kg/d.

As a preferable technical scheme of the invention, the medicine also comprises pharmaceutically acceptable auxiliary materials.

Preferably, the pharmaceutically acceptable auxiliary materials comprise any one or a combination of at least two of diluents, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, antioxidants, bacteriostats or buffers.

Preferably, the dosage form of the medicament comprises any one of suspension, granules, capsules, powder, tablets, emulsions, solutions, dripping pills, injections, suppositories, enemas, aerosols, patches or drops or the combination of at least two of the suspension, the granules, the capsules, the powders, the tablets, the emulsions, the solutions, the pills, the injections, the suppositories, the enemas, the aerosols, the patches or the drops.

Preferably, the administration route of the medicament comprises any one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration or a combination of at least two of the same.

In a second aspect, the present invention also provides the use of loratadine or ebastine for the preparation of an inhibitor of SARS-CoV-2 inflammatory factor, said SARS-CoV-2 inflammatory factor comprising an inflammatory factor induced by SARS-CoV-2spike-RBD protein.

In a third aspect, the invention provides the use of loratadine or ebastine in the preparation of mast cell degranulation inhibitors.

In a fourth aspect, the present invention also provides a pharmaceutical composition for treating COVID-19, said pharmaceutical composition comprising loratadine and/or ebastine;

preferably, the pharmaceutical composition further comprises an antiviral drug.

In the invention, the pharmaceutical composition can be a single compound preparation, and can also be prepared by combining preparations prepared from all components.

Likewise, the individual formulations of the pharmaceutical composition may be administered simultaneously or sequentially.

The recitation of numerical ranges herein includes not only the above-recited values, but also any values between any of the above-recited numerical ranges not recited, and for brevity and clarity, is not intended to be exhaustive of the specific values encompassed within the range.

Compared with the prior art, the invention has the beneficial effects that:

(1) the invention provides an application of loratadine or ibastine in preparing a medicament for preventing and/or treating COVID-19 inflammation. In vitro experiments, the lung epithelial cells A549 are cultured by using loratadine or ibastine to inhibit spike protein to stimulate the degranulation supernatant of LAD2, so that the inflammatory response of the A549 cells can be reduced, and further, the loratadine and ibastine are verified to reduce the inflammatory response caused by SARS-CoV-2 by inhibiting the degranulation of mast cells. In vivo experiments, the mouse model humanized with ACE2, C57BL/6N-ACE2^{em2(hACE2-WPRE,pgk-puro)/CCLA}It is further confirmed that loratadine and ibastine can inhibit inflammatory factor storm by inhibiting mast cell degranulation, and simultaneously prevent lung injury.

(2) The invention proves that the loratadine and ibastine can effectively inhibit the inflammatory factor storm caused by SARS-CoV-2 and have certain protection to tissue damage, so the loratadine and ibastine can be used as immunomodulators to reduce the COVID-19 inflammatory factor reaction; in addition, the composition is expected to be combined with antiviral drugs to treat severe infection, and in conclusion, the loratadine or ibastine combined antiviral drugs have important clinical application value in treating COVID-19.

Drawings

FIG. 1 is a graph comparing the degranulation of cells after 30 minutes and 2 hours of incubation for each experimental group and control group in example 1.

FIG. 2 is a bar graph showing the relative expression amounts of mRNAs of cytokines IL-6, IL-8, IL-1. beta., TNF. alpha., CCL20 and CCL5 in each of the experimental group and the control group in example 2, wherein I represents IL-6, II represents IL-8, III represents IL-1. beta., IV represents TNF. alpha., V represents CCL20, and VI represents CCL 5.

FIG. 3 is a graph showing the results of flow-based assay of the expression levels of cytokines IL-6, IL-8 and IL-1. beta. in each of the experimental group and the control group in example 2.

FIG. 4 is a toluidine blue staining pattern (scale 50 μm) of lung tissue of SARS-CoV-2 challenge group of example 3, wherein a ', b ', and c ' are magnified images of the regions indicated by a, b, and c, respectively.

FIG. 5 is a graph showing H & E staining of lung tissue of SARS-CoV-2 challenge group in example 3 (scale: 50 μm).

FIG. 6(A) is a toluidine blue staining pattern (scale 50 μm) of lung tissue of group Lor. + SARS-CoV-2 in example 3, wherein a' is an enlarged view of the region shown in a in the figure.

FIG. 6(B) is a H & E staining pattern (scale 50 μm) of lung tissue of group Lor. + SARS-CoV-2 in example 3, wherein a' is an enlarged view of the region indicated by a in the figure.

FIG. 7(A) is a toluidine blue staining pattern (scale 50 μm) of lung tissue of group Eba. + SARS-CoV-2 in example 3, wherein a' is an enlarged view of the region indicated by a in the figure.

FIG. 7(B) is an H & E staining pattern (scale 50 μm) of lung tissue of Eba. + SARS-CoV-2 group in example 3, wherein a' is an enlarged view of the region indicated by a in the figure.

FIG. 8(A) is a graph showing a comparison of the results of H & E staining of lung tissue damage in each of the experimental groups and the control group in example 3.

Fig. 8(B) is a graph comparing the statistics of the number of mast cells in toluidine blue staining of each experimental group and the control group in example 3.

FIG. 9 is a graph showing a comparison of the expression levels of the mRNAs of the cytokines IL-6, IL-8, IL-1. beta., TNF. alpha., IFN-. gamma., CRP, CCL20 and CCL5 in each of the experimental group and the control group in example 3, wherein I represents IL-6, II represents IL-8, III represents IL-1. beta., IV represents TNF. alpha., V represents CRP, VI represents IFN-. gamma., VII represents CCL20, and VIII represents CCL 5.

Detailed Description

The technical solutions of the present invention are further described in the following embodiments with reference to the drawings, but the following examples are only simple examples of the present invention and do not represent or limit the scope of the present invention, which is defined by the claims.

In the following examples, the main reagents and antibodies used were as follows:

spike-RBD protein (Genscript); ibastine (selelck); loratadine (selelck); anti-Avidin-FITC (Invitrogen); ReverTra qPCR RT Master Mix gDNA remove (Toyobo); SYBR qPCR Mix (Genestar);

immunofluorescence antibodies: alexa Fluor647-anti-IL-1 beta (Biolegend, JK1B-1), PE-anti-IL-6(BD, MQ2-6A3), Alexa Fluor488-anti-TNF alpha (BD, MAb11), BV421-anti-IL-8(BD, G265-8);

RPMI-1640 medium (GIBCO); fetal Bovine Serum (FBS) (Millipore); dimethyl sulfoxide (DMSO) (Sigma); paraformaldehyde Fixative (PFA) (Sigma); phosphate Buffer (PBS) (Hyclone); Triton-X100 (Sigma); trizol (invitrogen); toluidine blue (Sigma); FACS buffer (Fluorescence activated Cell licensing, FACS); leukocyte activation reagents (BD, 550583); fixative/rupture fluid (BD, 554722).

The rest of the reagents and consumables are purchased from conventional reagent manufacturers in the field unless otherwise specified.

In the following examples, unless otherwise specified, all experimental methods and technical means are those conventional in the art. Wherein, the related primer sequences are as follows:

human source: IL-6-F (SEQ ID NO.1), 5'-CAGACAGCCACTCACCTCTTCAG-3';

IL-6-R(SEQ ID NO.2)，5′-CAGCCATCTTTGGAAGGTTCAG-3′；

TNF-α-F(SEQ ID NO.3)，5′-CCCAGGCAGTCAGATCATCTTC-3′；

TNF-α-R(SEQ ID NO.4)，5′-GTGAGGAGCACATGGGTGGAG-3′；

MMP9-F(SEQ ID NO.5)，5′-CCTTCTACGGCCACTACTGTGC-3′；

MMP9-R(SEQ ID NO.6)，5′-GCCAGTACTTCCCATCCTTGAAC-3′；

GAPDH-F(SEQ ID NO.7)，5′-ATCCCATCACCATCTTCCAGG-3′；

GAPDH-R(SEQ ID NO.8)，5′-CCTTCTCCATGGTGGTGAAGAC-3′；

IL-1β-F(SEQ ID NO.9)，5′-CGTCAGTTGTTGTGGCCATGGA-3′；

IL-1β-R(SEQ ID NO.10)，5′-GAGCGTGCAGTTCAGTGATCGTA-3′；

IL-8-F(SEQ ID NO.11)，5′-CTGATTTCTGCAGCTCTGTGTGA-3′；

IL-8-R(SEQ ID NO.12)，5′-GGTCCAGACAGAGCTCTCTTCCA-3′；

CCL20-F(SEQ ID NO.13)，5′-GCTGCTTTGATGTCAGTGCT-3′；

CCL20-R(SEQ ID NO.14)，5′-TGTCACAGCCTTCATTGGC-3′；

CCL5-F(SEQ ID NO.15)，5′-ACCACACCCTGCTGCTTTG-3′；

CCL5-R(SEQ ID NO.16)，5′-GCGGTTCTTTCGGGTGACA-3′；

mouse source: IL-6-F (SEQ ID NO.17), 5'-CTTCCATCCAGTTGCCTTCTTG-3';

IL-6-R(SEQ ID NO.18)，5′-AATTAAGCCTCCGACTTGTGAAG-3′；

TNF-α-F(SEQ ID NO.19)，5′-CAGACCCTCACACTCAGATCATCT-3′；

TNF-α-R(SEQ ID NO.20)，5′-CCTCCACTTGGTGGTTTGCTA-3′；

IL-1β-F(SEQ ID NO.21)，5′-CTTTCAGAGGCCAGAGAGTCC-3′；

IL-1β-R(SEQ ID NO.22)，5′-TCCCTGTAGTGACAGCACCT-3′；

IL-8-F(SEQ ID NO.23)，5′-CGGCAATGAAGCTTCTGTAT-3′；

IL-8-R(SEQ ID NO.24)，5′-CCTTGAAACTCTTTGCCTCA-3′；

INFγ-F(SEQ ID NO.25)，5′-ATGAACGCTACACACTGCATC-3′；

INFγ-R(SEQ ID NO.26)，5′-CCATCCTTTTGCCAGTTCCTC-3′；

GAPDH-F(SEQ ID NO.27)，5′-TGCACCACCAACTGCTTAG-3′；

GAPDH-R(SEQ ID NO.28)，5′-GATGCAGGGATGATGTTC-3′；

CRP-F(SEQ ID NO.29)，5′-CAGAGATTCCTGAGGCTCCAACA-3′；

CRP-R(SEQ ID NO.30)，5′-AGTCACCGCCATACGAGTCCTG-3′；

CCL20-F(SEQ ID NO.31)，5′-AAGACAGATGGCCGATGAAG-3′；

CCL20-R(SEQ ID NO.32)，5′-AGGTTCACAGCCCTTTTCAC-3′；

CCL5-F(SEQ ID NO.33)，5′-GTGCCCACGTCAAGGAGTAT-3′；

CCL5-R(SEQ ID NO.34)，5′-GGGAAGCTATACAGGGTCA-3′；

the qRT-PCR detection viral load primers are as follows:

N-F(SEQ ID NO.35)，5′-GGGGAACTTCTCCTGCTAGAAT-3′；

N-R(SEQ ID NO.36)，5′-CAGACATTTTGCTCTCAAGCTG-3′；

the probe sequence (SEQ ID NO.37) is:

5′-FAM-TTGCTGCTGCTTGACAGATT-TAMRA-3′。

all animal experiments were approved by the welfare and ethics committees of experimental animals of kunming animal institute, national academy of sciences, Guangzhou institute of biology and Chinese academy of sciences. All SARS-CoV-2 infection experiments were performed in a biosafety class III laboratory.

Example 1

This example is presented to demonstrate that loratadine or ibastine is able to inhibit the degranulation of LAD 2.

(1) Will be 5X 10⁵LAD2 cells/well were plated in 24-well cell culture plates and incubated for 20 hours with ebastine (3. mu.g/mL) and loratadine (5. mu.g/mL), respectively;

(2) stimulating cells (namely Eba. + RBD and Lor. + RBD) by using SARS-CoV-2sipke-RBD (10 mu g/mL), stimulating cells by using spike-packaged lentiviruses (namely Eba. + pseudoviruses and Lor. + pseudoviruses) and collecting cells at 30 minutes and 2 hours respectively;

(3) fixing the cells with 4% Paraformaldehyde (PFA) at room temperature for 30 minutes, washing with PBS for 3 times, incubating with anti-Avidin-FITC at 4 ℃ in the dark for 1 hour, washing with PBS for 3 times, and performing flow detection;

the resulting spike-RBD protein or spike-packaged lentivirus induced mast cell degranulation as shown in FIG. 1, whereas the level of degranulation of cells after treatment with ibastine or loratadine was significantly lower than that stimulated by spike-RBD alone, indicating that the use of loratadine or ibastine alone inhibited the degranulation induced by spike-RBD.

Example 2

This example is presented to demonstrate the inhibitory effect of loratadine and ibastine on the inflammatory factor storm induced by SARS-CoV-2.

(2) after stimulating the cells with sipke-RBD (10 μ g/mL) for 2 hours, cell supernatants were collected for culturing a549 cells, yielding Eba. + RBD and Lor. + RBD groups;

(3) a549 cells were collected 24 hours later and cytokine expression was determined by quantifying mRNA production and intracellular immunostaining with specific antibodies, respectively;

(4) at mRNA level, Trizol lyses cells to extract RNA, reverse transcribes the RNA into cDNA by using ReverTra qPCR RT Master Mix gDNA remover, and detects the expression of IL-6, IL-8, TNF alpha and IL-1 beta on a Bio-rad CFX96 PCR instrument by using Thunderbird SYBR qPCR Mix;

the sequence of the primer is shown as SEQ ID NO. 1-16;

the results of stimulating the degranulation of LAD2 with ibastine (Eba. + RBD group) or loratadine (Lor. + RBD group) by inhibiting the Spike-RBD protein and culturing A549 cells with the supernatant are shown in FIG. 2, and it can be seen that the mRNA expression levels of IL-6 (panel I), IL-8 (panel II), TNF α (panel III), IL-1 β (panel IV), CCL20 (panel V) and CCL5 (panel VI) are significantly increased on average;

wherein, the experimental group and the control group comprise: culture in Medium blank Medium (Medium), culture in Medium containing sipke-RBD (RBD Medium group), culture in LAD2 supernatant without any treatment (LAD2 Medium group), culture in LAD2 cell supernatant after sipke-RBD treatment (RBD group), pretreatment with ibastine, culture in LAD2 cell supernatant after sipke-RBD treatment (Eba. + RBD group), pretreatment with loratadine, and culture in LAD2 cell supernatant after sipke-RBD treatment (Lor. + RBD group).

(5) In the intracellular immunostaining experiment, a leukocyte activation reagent and a protein transport inhibitor are firstly added into A549 cells for 6 hours, then the cells are fixed by using a fixed membrane-breaking liquid at 4 ℃ for 20 minutes, fluorescent antibodies are dyed overnight at 4 ℃ in a dark place, and after washing, the cells are resuspended in a FACS buffer solution for flow cytometry analysis.

The results are shown in FIG. 3, in which the Isotype group stained with the control antibody to eliminate background staining due to non-specific binding of the antibody; the medium group is a control group, and the same target antibody is used in the experimental group; protein expression of IL-6, IL-8 and IL-1 β were mean-significantly down-regulated in the Eba. + RBD and Lor. + RBD groups compared to the RBD group.

The results also demonstrate that, at the cellular level, ibastine and loratadine are able to inhibit inflammatory factor production by inhibiting mast cell degranulation.

Example 3

In this example, an ACE2 humanized mouse model was used to verify the inhibitory effect of loratadine and ibastine on the inflammatory factor storm induced by SARS-CoV-2 and the protective effect on lung injury.

(1) The ACE2 humanized mouse model is C57BL/6N-ACE2^{em2(hACE2-WPRE,pgk-puro)/CCLA}The mice are intraperitoneally injected with ebastine (5mg/kg) or loratadine (10mg/kg) one day before the mice are attacked by the poison at a dose of 5 x10⁶TCID50；

(2) Injecting the same dose of ebastine or loratadine every day thereafter to day four;

(3) on day 5 post-infection, the mice were anesthetized and euthanized and their lungs were subjected to RNA extraction and histopathological staining (H & E, toluidine blue), respectively; the method comprises the following steps of detecting the expression of IL-6, IL-8, TNF alpha, IL-1 beta, INF gamma, CRP, CCL20 and CCL5 by qPCR, wherein the sequences of primers and probes are shown in SEQ ID NO. 17-37;

pathological lung lesions and mast cell degranulation were detected by H & E and toluidine blue staining.

As shown in the histopathological results of fig. 4, the mice showed massive degranulation of mast cells in the lungs after challenge and more severe lung injury; wherein, the lung injury position is shown in the position shown by the arrow in fig. 5, which comprises: thickening of alveolar spaces, bleeding, mucosal exfoliation, formation of transparent membranes, infiltration of inflammatory cells, and fibrous exudation.

FIGS. 6(A) and 6(B) show the results of the staining of Eba. + SARS-CoV-2 group, FIGS. 7(A) and 7(B) show the results of the staining of Lor. + SARS-CoV-2 group, the degree of lung tissue damage in H & E staining of each group was scored and the number of mast cells in toluidine blue staining was counted;

the scoring and statistical results are shown in fig. 8(a) and fig. 8(B), and it can be seen that the mouse lung tissue damage degree and mast cell degranulation of Lor ++ SARS-CoV-2 and Eba ++ SARS-CoV-2 groups are significantly lower than those of SARS-CoV-2 challenge group.

In addition, the results of quantitative detection of (RT-) RNA are shown in FIG. 9, and after SARS-CoV-2 infection, mRNA expression levels of IL-6 (panel I), IL-8 (panel II), TNF alpha (panel III), IL-1 beta (panel IV), CRP (panel V), INF gamma (panel VI), CCL20 (panel VII) and CCL5 (panel VIII) were significantly up-regulated on a water-average basis, indicating that severe inflammatory factor storm occurred in the lung after mouse challenge; after ebastine or loratadine are used, the expression of IL-6, IL-8, TNF alpha, IL-1 beta, INF gamma, CRP (C-reactive protein), CCL20 and CCL5 induced by SARS-CoV-2 can be inhibited.

Thus, the results of the examples of the present invention show that the results are shown at C57BL/6N-Ace2^{em2(hACE2-WPRE,pgk-puro)/CCLA}In a mouse model, ibastine and loratadine can inhibit the generation of inflammatory factors induced by SARS-CoV-2 and prevent lung injury induced by SARS-CoV-2 by inhibiting mast cell degranulation.

In conclusion, the invention proves that loratadine and ibastine can inhibit the generation of inflammatory factors caused by SARS-CoV-2, protect lung and prevent injury in vivo and in vitro; the result indicates that the loratadine and ibastine can be used as immunomodulators, reduce the COVID-19 inflammatory factor reaction, are expected to be combined with antiviral drugs to treat severe infection, and have important clinical application value in the treatment of COVID-19.

The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are within the scope and disclosure of the present invention.

Sequence listing

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atgaacgcta cacactgcat c 21

<210> 26

<211> 21

<212> DNA

<213> Artificial Synthesis ()

<400> 26

ccatcctttt gccagttcct c 21

<210> 27

<211> 19

<212> DNA

<213> Artificial Synthesis ()

<400> 27

tgcaccacca actgcttag 19

<210> 28

<211> 18

<212> DNA

<213> Artificial Synthesis ()

<400> 28

gatgcaggga tgatgttc 18

<210> 29

<211> 23

<212> DNA

<213> Artificial Synthesis ()

<400> 29

cagagattcc tgaggctcca aca 23

<210> 30

<211> 22

<212> DNA

<213> Artificial Synthesis ()

<400> 30

agtcaccgcc atacgagtcc tg 22

<210> 31

<211> 20

<212> DNA

<213> Artificial Synthesis ()

<400> 31

aagacagatg gccgatgaag 20

<210> 32

<211> 20

<212> DNA

<213> Artificial Synthesis ()

<400> 32

aggttcacag cccttttcac 20

<210> 33

<211> 20

<212> DNA

<213> Artificial Synthesis ()

<400> 33

gtgcccacgt caaggagtat 20

<210> 34

<211> 19

<212> DNA

<213> Artificial Synthesis ()

<400> 34

gggaagctat acagggtca 19

<210> 35

<211> 22

<212> DNA

<213> Artificial Synthesis ()

<400> 35

ggggaacttc tcctgctaga at 22

<210> 36

<211> 22

<212> DNA

<213> Artificial Synthesis ()

<400> 36

cagacatttt gctctcaagc tg 22

<210> 37

<211> 20

<212> DNA

<213> Artificial Synthesis ()

<400> 37

ttgctgctgc ttgacagatt 20

Claims

1. Application of loratadine or ebastine in preparing medicine for preventing and/or treating COVID-19 inflammation is provided.

2. The use according to claim 1, wherein the loratadine is used in an amount of 0.1-10 mg/kg/d, preferably 0.2-0.5 mg/kg/d.

3. The use according to claim 1 or 2, wherein ebastine is used in a dose of 0.1-20 mg/kg/d, preferably 0.1-1 mg/kg/d.

4. The use according to any one of claims 1 to 3, wherein the medicament further comprises a pharmaceutically acceptable excipient.

5. The use according to claim 4, wherein the pharmaceutically acceptable excipient comprises any one or a combination of at least two of diluents, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, tonicity adjusting agents, surfactants, coating materials, antioxidants, bacteriostats or buffers.

6. The use according to any one of claims 1 to 5, wherein the medicament is in a dosage form selected from one or a combination of at least two of suspension, granules, capsules, powders, tablets, emulsions, solutions, drop pills, injections, suppositories, enemas, aerosols, patches and drops.

7. The use according to any one of claims 1 to 6, wherein the route of administration of the medicament comprises any one or a combination of at least two of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration.

8. Use of loratadine or ebastine for the preparation of an inhibitor of SARS-CoV-2 inflammatory factor, wherein the SARS-CoV-2 inflammatory factor comprises an inflammatory factor induced by SARS-CoV-2spike-RBD protein.

9. Application of loratadine or ebastine in preparing mast cell degranulation inhibitor is provided.

10. A pharmaceutical composition for treating COVID-19, wherein the pharmaceutical composition comprises loratadine and/or ebastine;

preferably, the pharmaceutical composition further comprises an antiviral drug.