CN115068454A - Application of salmeterol medicine for preventing and treating coronavirus infection - Google Patents

Abstract

The invention belongs to the technical field of medicines, and relates to a medicinal application of an antiasthmatic medicament Salmeterol (Salmeterol) in preventing and treating coronavirus infection. The innovation of the invention is that the medicine is relocated: the research finds that salmeterol has the function of inhibiting the replication of a novel coronavirus (SARS-CoV-2); has effects in inhibiting replication of coronavirus HCoV-OC43 and HCoV-229E. The invention provides a scientific experimental basis for expanding the clinical indications of salmeterol and provides a feasible and effective treatment scheme for clinically preventing and treating coronavirus infectious diseases.

Description

Application of salmeterol medicine for preventing and treating coronavirus infection

Technical Field

The invention belongs to the technical field of medicines, and relates to a medicinal application of salmeterol in preventing and treating coronavirus infectious diseases.

Background

The prevention and treatment of these viruses presents challenges due to the lack of effective therapeutic modalities for the prevention and treatment of the novel coronavirus (SARS-CoV-2), the common coronavirus (α -type, β -type coronavirus). Therefore, there is a need to find drugs with a broad spectrum of anti-coronavirus effects.

Disclosure of Invention

The present invention relates to methods and results for the prevention and treatment of novel, common coronavirus infections according to formula I.

It has been found that the antiasthmatic drug Salmeterol (Salmeterol), herein designated Compound I (LG0014), exhibits antiviral activity against the novel coronaviruses (SARS-CoV-2), coronaviruses (HCoV-OC43) and coronaviruses (HCoV-229E).

The technical problem to be solved by the invention is to provide the effect of salmeterol or pharmaceutically acceptable salt thereof in preparing medicaments for preventing and treating coronavirus infectious diseases.

Specifically, in order to solve the technical problem of the present invention, the following technical scheme is adopted:

the first aspect of the technical proposal of the invention provides the application of salmeterol shown as a structural formula I or pharmaceutically acceptable salt thereof in preparing the medicaments for preventing or treating coronavirus infection,

the salmeterol pharmaceutically acceptable salt comprises pharmaceutically acceptable organic salt or inorganic salt, wherein the organic salt comprises sulfonate, carboxylate, amino acid salt and fatty acid salt, and the inorganic salt comprises hydrochloride, bromate, iodate, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate and nitrate. The sulfonate comprises alkyl sulfonate containing 1-15 carbon atoms, benzene sulfonate, p-toluene sulfonate, o-toluene sulfonate and m-toluene sulfonate; carboxylates including tartrate, maleate, fumarate, citrate, malate, cinnamate, benzoate, malonate, succinate, glutarate, adipate, pamoate, and lactate; amino acid salts include glutamate and aspartate; the fatty acid salt comprises a long chain fatty acid salt having 2 to 18 carbon atoms.

Wherein the coronavirus comprises novel coronavirus SARS-CoV-2 and novel coronary pneumonia COVID-19 induced by the coronavirus SARS-CoV-2.

The coronavirus infection comprises infectious diseases induced by HCoV-OC43 and HCoV-229E coronavirus.

In a second aspect of the technical scheme of the invention, the invention provides an application of a pharmaceutical composition in preparing a medicament for preventing or treating coronavirus infection, wherein the pharmaceutical composition comprises salmeterol shown in a structural formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

general description

The invention is first described in detail with reference to the noun terms that are used to facilitate a better understanding of the invention.

The term "antiviral drug" as used herein refers to a drug (compound, crude extract or biological agent) effective in inhibiting the formation or replication of a virus in a human body, including but not limited to drugs that interfere with host proteins or viral mechanisms necessary for the virus during the formation or replication of the human body.

"prevention" (or prevention) refers to any treatment that prevents the disease or disorder from further developing. The term "preventing" also includes the use of a therapeutically effective amount of a compound or composition of the invention prior to exposure of an individual to a virus (i.e., pre-exposure prevention) to prevent the symptoms of disease from developing and/or to prevent the virus from reaching detectable levels in a sample of the patient's virus.

"treating" (or treating) means reversing, alleviating, or inhibiting one or more symptoms of the disease for which prevention is indicated. In certain embodiments, "treatment" refers to a compound or composition according to the present invention to reduce or eliminate symptoms of viral infection and/or reduce the viral load of a patient's viral sample.

The term "pharmaceutical composition" refers to a pharmaceutical composition prepared by mixing the compound of the present invention with adjuvants or other active ingredients in any ratio.

The compound names provided above are named according to literature, and one skilled in the art will appreciate that other recognized naming systems and symbols can be used to name or identify the compound structure. For example, a compound may be named or identified by a common name, a system or non-system name. Commonly accepted nomenclature systems and symbols in the chemical arts include, but are not limited to, the Chemical Abstracts Service (CAS) and the International Union of Pure and Applied Chemistry (IUPAC). Thus, the structure of compound formula I (LG0014) provided above is named 4- (1-hydroxy-2- ((6- (4-phenylbutoxy) hexyl) amino) ethyl) -2- (hydroxymethy) phenol using the ChemBioDraw Ultra name.

The beneficial technical effects are as follows: salmeterol is a selective beta 2 receptor agonist, has a bronchodilating effect and is a drug which is already on the market. The research shows that the salmeterol has broad-spectrum antiviral effect, including novel coronavirus (SARS-CoV-2), coronavirus HCoV-OC43 and HCoV-229E, and has good clinical application prospect.

Drawings

FIG. 1 of formula I, LG0014 ¹ H NMR spectrum

FIG. 2 of formula I, LG0014 ¹³ C NMR spectrum

FIG. 3H-COSY-MOF map of formula I, LG0014

FIG. 4 Mass Spectroscopy of formula I, LG0014

FIG. 5 shows the pharmacodynamic curves of LG0014 against neocoronaviruse-induced cytopathic effect

Detailed Description

Experimental example 1 evaluation of salmeterol action against novel coronavirus (SARS-CoV-2)

Cell: vero cells.

Virus strain: C-Tan-nCoV strain 01, the viral infection corresponds to 100 TCID.

Experimental materials and methods:

(1) sample treatment: the sample concentration in DMSO, 10mg/ml, was used as stock solution. The drug was diluted with a virus culture maintenance medium just before use, the first concentration was 100ug/ml, and then 5 dilutions were made with the culture medium, each at 5 dilutions. The positive drug is Reidesvir.

(2) A therapeutic administration mode is adopted, the virus and the cells are incubated for 2h, then virus liquid is removed, the medicine diluted by the maintenance liquid is added, and the culture is continued for 48 h. And simultaneously setting a medicine control group and a virus control group.

(3) Viral nucleic acid assay: after the cells are treated by the drug for 48 hours, 100 mul of cell culture supernatant is respectively taken to extract nucleic acid, an rRT-PCR experiment (ORF1ab) is carried out, the Ct value of the novel coronavirus nucleic acid in the cell culture supernatant is measured, and the virus inhibition rate is calculated.

(4) Calculating the EC of the sample ₅₀ 。

The experimental results are shown in Table 1, and the pharmacodynamic graph is shown in FIG. 5.

The results show that salmeterol has significant anti-new coronavirus effect, EC ₅₀ 0.88ug/ml, similar to the positive drug Reddesivir (Remdesivir), provides a new protocol for clinical treatment of new coronary pneumonia.

TABLE 1 inhibition of the cytopathic effect induced by the novel coronavirus C-Tan-nCoV strain 01 (SARS-CoV-2) by salmeterol on Vero cells in vitro

^a TC ₅₀ Average 50% cytotoxic compound concentration

^b EC ₅₀ Concentration of average 50% inhibition

^c SI selection index, TC ₅₀ /EC ₅₀ .

Experimental example 2 evaluation of Activity of Salmeterol against coronavirus HCoV-OC43

H460 cells were used as virus hosts, and the extent of cytopathic effect (CPE) of the sample on the virus-induced cells was determined. Virus strain: the coronavirus HCoV-OC43 was stored at-80 ℃. Sample treatment: samples were made up to stock solution with DMSO concentration of 10mg/ml and 3-fold dilutions were made in culture medium, 8 dilutions each. Positive control drug: ribavirin (RBV), Hubei Tian Yao pharmaceutical GmbH (batch No. 31712252).

The test method comprises the following steps: inoculating H460 cells into 96-well culture plate, and placing 5% CO ₂ And cultured at 37 ℃. Infection after 24 hours is about 100TCID ₅₀ Adding maintenance solution containing samples with different dilutions and positive control drug, setting cell control hole and virus control hole, and 5% CO ₂ And cultured at 37 ℃. Observing the pathological change degree (CPE) of each group of cells when the pathological change degree (CPE) of the virus control group reaches 4+, and respectively calculating the half Toxic Concentration (TC) of the sample to the cells by using a Reed-Muench method ₅₀ ) And half inhibitory concentration (EC) against virus ₅₀ ). The results are shown in Table 2.

The results show that the EC of salmeterol against coronavirus HCoV-OC43 ₅₀ The activity of the compound is equivalent to that of ribavirin which is a positive drug, namely 4.31ug/ml, which indicates that the salmeterol has remarkable effect of resisting coronavirus HCoV-OC 43.

TABLE 2 salmeterol inhibition of coronavirus HCoV-OC 43-induced cytopathic effects on H460 cells in vitro

^a TC ₅₀ Average 50% cytotoxic compound concentration

^b EC ₅₀ Concentration of average 50% inhibition

^c SI selection index, TC ₅₀ /EC ₅₀ .

Experimental example 3 evaluation of Activity against coronavirus HCoV-229E

Huh7 cells were used as virus hosts to determine the extent of cytopathic effect (CPE) of the samples in inhibiting the virus. Virus strain: the coronavirus HCoV-229E was stored at-80 ℃. Samples were made up as stock solutions in DMSO at a concentration of 10 mg/ml. The culture medium was further diluted 3-fold, 8 dilutions each. Positive control drug: ribavirin (RBV), Hubei Tian Yao pharmaceutical Co., Ltd. (batch No. 31712252).

The test method comprises the following steps: inoculating Huh7 cells into 96-well culture plate, and placing 5% CO ₂ And cultured at 37 ℃. 2Infection after 4 hours was about 100TCID ₅₀ Adding maintenance solution containing samples with different dilutions and positive control drug, setting cell control hole and virus control hole, and 5% CO ₂ And cultured at 37 ℃. Observing the pathological change degree (CPE) of each group of cells when the pathological change degree (CPE) of the virus control group reaches 4+, and respectively calculating the half Toxic Concentration (TC) of the sample to the cells by using a Reed-Muench method ₅₀ ) And half inhibitory concentration (EC) against virus ₅₀ ). The results are shown in Table 3.

The results show that the EC of salmeterol against coronavirus HCoV-229E ₅₀ The activity of the compound is equivalent to that of ribavirin which is a positive drug, namely 1.85ug/ml, which indicates that the salmeterol has obvious effect of resisting coronavirus HCoV-229E.

TABLE 3 salmeterol inhibition of coronavirus HCoV-229E-induced cytopathic effects on Huh7 cells in vitro

^a TC ₅₀ Average 50% cytotoxic compound concentration

^b EC ₅₀ Concentration of average 50% inhibition

^c SI selection index, TC ₅₀ /EC ₅₀ .

In conclusion, salmeterol has the effects of relieving asthma, resisting novel coronavirus SARS-CoV-2 and resisting coronavirus HCoV-OC43 and HCoV-229E, has broad-spectrum coronavirus infection resistance, and is expected to become an effective treatment medicament for preventing and treating coronavirus infectious diseases. The invention lays a scientific experimental foundation for enlarging the clinical indications of salmeterol.

The invention belongs to the relocation of a medicine of Samette which is an antiasthmatic medicine on the market. The broad-spectrum anti-coronavirus effect of Samite provides a feasible and effective treatment scheme for clinically preventing and treating coronavirus infectious diseases.

Claims (6)

1. The application of the salmeterol shown in the formula I or the pharmaceutically acceptable salt thereof in preparing the medicaments for preventing and/or treating the coronavirus infection,

2. the use according to claim 1, wherein the pharmaceutically acceptable salt comprises a pharmaceutically acceptable organic salt or inorganic salt, wherein the organic salt comprises a sulfonate, a carboxylate, an amino acid salt, and a fatty acid salt, and the inorganic salt comprises a hydrochloride, a bromate, an iodate, a sulfate, a hydrogen sulfate, a phosphate, a hydrogen phosphate, a dihydrogen phosphate, and a nitrate.

3. Use according to claim 2, characterized in that said sulfonates comprise alkylsulfonates containing 1-15 carbon atoms, benzenesulfonates, p-toluenesulfonates, o-toluenesulfonates, m-toluenesulfonates; carboxylates include tartrate, maleate, fumarate, citrate, malate, cinnamate, benzoate, malonate, succinate, glutarate, adipate, pamoate, and lactate; amino acid salts include glutamate, aspartate; the fatty acid salt comprises a long chain fatty acid salt having 2 to 18 carbon atoms.

4. Use according to claim 1, characterized in that the coronavirus-neocoronavirus SARS-CoV-2 and infectious diseases induced by it are prevented or treated.

5. Use according to claim 1, characterized in that the coronary diseases-HCoV-OC 43, HCoV-229E and infectious diseases induced by them-are prevented or treated.

6. A pharmaceutical composition is used for preparing a medicament for preventing or treating coronavirus infectious diseases, and is characterized in that the pharmaceutical composition comprises salmeterol shown as a structural formula I or pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

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