CN115813893A - Application of naphthyl skeleton compound in preparing coronavirus 3CL protease inhibitor - Google Patents

Abstract

The invention provides application of naphthyl skeleton compounds in preparing coronavirus 3CL protease inhibitors, wherein the naphthyl skeleton compounds can effectively inhibit 3CL ^pro The activity of (3) has high therapeutic index and low half effective concentration, further inhibits the activity of coronavirus, and can be potentially used for developing anti-coronavirus medicines.

Description

Application of naphthyl skeleton compound in preparing coronavirus 3CL protease inhibitor

Technical Field

The invention belongs to the field of medicines, and relates to application of a compound taking naphthalene as a framework in preparation of a coronavirus 3CL protease inhibitor.

Background

Research on new coronaviruses, which mainly include novel coronavirus pneumonia (SARS-CoV-2), severe acute respiratory syndrome-associated coronavirus (SARS-CoV) and middle east syndrome-associated coronavirus (MERS-CoV), mainly positive-strand single-stranded RNA virus of mantle (envelope), and have entered the athermalization stage, has been conducted, and the diameter thereof is about 80 to 120nm. An attractive drug target in coronaviruses is the major protease (M) ^pro ，3CL ^pro ) Since it plays a crucial role in the processing of polyproteins translated from viral RNA. 3CL ^pro Have been shown to be drug discovery targets for SARS, MERS and SARS-CoV-2 coronavirus. Most of the functional proteins (non-structural proteins) of coronaviruses are encoded by ORF1ab gene, translated into a polyprotein (7096 aa) and then 3CL ^pro Cleavage into multiple active proteins such as the viral replication protein RdRp. As an important non-structural protein in coronaviruses, the protein has similar cleavage site specificity with 3CL protease of microRNA virus. 3CL ^pro Is composed of 306 amino acids, a cysteine protease of about 33kDa (much smaller than S protein) that specifically recognizes and cleaves 11 cleavage sites of the nonstructural protein NSP4-NSP16 to release the other nonstructural proteins of coronavirus, and 3CL therefor ^pro The method is a very hot target in the research of the development of the current new crown drugs. However, no effective anti-neocoronal drug exists at present.

Disclosure of Invention

The invention provides application of naphthyl skeleton compounds in preparation of coronavirus 3CL protease inhibitors, and is expected to solve the problem that no anti-corona compound exists at presentThe current state of the art for the treatment of viroids, compounds with naphthalene as backbone, are effective in inhibiting 3CL ^pro Thus inhibiting the activity of coronavirus, and can be potentially used for developing anti-coronavirus medicines.

The invention is realized by the following technical scheme:

the invention provides application of a naphthyl skeleton compound in preparing a coronavirus 3CL protease inhibitor.

The invention also provides application of the naphthyl skeleton compound in preparing medicaments for preventing and/or treating diseases caused by coronavirus.

Preferably, the naphthyl skeleton compound is a pharmaceutically acceptable salt, solvate, enantiomer, diastereoisomer, tautomer or mixture thereof in any proportion.

Preferably, the naphthyl skeleton compound is one of the following compounds:

preferably, the naphthyl skeleton compound is 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde or 1,4-dimethoxy-2-naphthaldehyde.

Preferably, the coronavirus is MERS-CoV, SARS-CoV or SARS-CoV-2.

Preferably, the coronavirus-caused disease is MERS-CoV, SARS-CoV-2-caused infectious disease or a complication thereof.

Preferably, the inhibitor or the medicament is an orally administered tablet, capsule, granule, syrup, premix or pellet, or a liniment or injection administered in a non-oral manner.

Preferably, the inhibitor is a naphthyl backbone compound used alone or a combination of the naphthyl backbone compound and other coronavirus 3CL protease inhibitors.

Preferably, the medicament is a naphthyl skeleton compound used alone or a composition of the naphthyl skeleton compound and other medicaments for preventing and/or treating diseases caused by coronavirus.

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

the invention proves that the naphthyl skeleton compound is applied to coronavirus 3CL for the first time ^pro The therapeutic index is high, and the half effective concentration is low; and the anti-virus effect can be effectively increased by using a compound taking naphthalene as a framework before infection. The naphthyl skeleton compound can be used as an effective medicament for treating coronavirus infection.

Drawings

FIG. 1 is a graph of the inhibition of SARS-CoV 3CL protease by 2,3-dimethoxy-1-naphthaldehyde in accordance with the present invention;

FIG. 2 is the inhibition of SARS-CoV 3CL protease by 2-hydroxy-7-methoxy-1-naphthaldehyde in accordance with the present invention;

FIG. 3 is a graph of the inhibition of SARS-CoV 3CL protease by 1,4-dimethoxy-2-naphthaldehyde in accordance with the present invention;

FIG. 4 shows the inhibition of SARS-CoV-2 3CL protease by 2,3-dimethoxy-1-naphthaldehyde in accordance with the present invention;

FIG. 5 shows the inhibition of SARS-CoV-2 3CL protease by 2-hydroxy-7-methoxy-1-naphthaldehyde in the present invention;

FIG. 6 shows the inhibition of SARS-CoV-2 3CL protease by 1,4-dimethoxy-2-naphthaldehyde in the present invention.

Detailed Description

For a further understanding of the invention, reference will now be made to the following examples, which are provided to illustrate further features and advantages of the invention, and are not intended to limit the scope of the invention as set forth in the following claims.

The following sources of raw materials are illustrative

2,3-dimethoxy-1-naphthaldehyde (CAS: 56252-09-6), 2-hydroxy-7-methoxy-1-naphthaldehyde (CAS: 75965-66-1), 1,4-dimethoxy-2-naphthaldehyde (CAS: 75965-83-2).

Example 1

2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde, and 1,4-dimethoxy-2-naphthaldehyde are used for in vitro enzyme activity inhibition detection of SARS-CoV 3CL protease.

The inhibition of SARS-CoV 3CL protease activity by 3 compounds was evaluated using the fluorescence resonance energy transfer method (each compound was tested in triplicate).

Weighing appropriate amount of the above compounds, preparing a solution with appropriate concentration gradient with DMSO, mixing 5 μ L of the prepared solution with 91 μ L of Assay Reagent (Assay Buffer: SARS-CoV M) ^pro /3CL ^pro =90:1, purchased from shanghai bi yunnan biotechnology limited) were sequentially added to black 96-well plates, mixed, incubated at 37 ℃ for 10 minutes in the dark, and 4 μ L of Substrate (100 μ M Dabcyl-KTSAVLQSGFRKME-Edans, purchased from shanghai bi yunnan biotechnology limited) was rapidly added to each well, mixed. After incubation for 5min at 37 ℃ in the dark, the signal tends to be stable, fluorescence measurement is carried out within 5-30min by using a multifunctional microplate reader (Serimei Feishell science and technology Co., ltd., varioskan Flash) and the inhibition percentage of the sample is calculated, wherein the excitation wavelength is 340nm and the emission wavelength is 490nm. Assay Reagent without compound was used as 100% enzyme activity control, without SARS-CoV M ^pro The Assay Buffer of/3 CL protease is a blank control, and the rest of the treatment methods are the same. IC of the samples was calculated using GraphPad Prism software for non-linear regression analysis ₅₀ The value is obtained.

The experimental results are shown in Table 1, and examples 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde, 1,4-dimethoxy-2-naphthaldehyde all have inhibitory activity against SARS-CoV 3CL protease, wherein 2,3-dimethoxy-1-naphthaldehyde has strong inhibitory effect against SARS-CoV 3CL protease, and IC is IC ₅₀ The values were all around 20. Mu.M. FIGS. 1-3 show the inhibition of SARS-CoV 3CL protease by 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde, 1,4-dimethoxy-2-naphthaldehyde, respectively.

TABLE 1 inhibition ratio of SARS-CoV 3CL protease (%)

Example 2

2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde, and 1,4-dimethoxy-2-naphthaldehyde are used for in vitro enzyme activity inhibition detection of SARS-CoV-2 3CL protease.

The inhibition of SARS-CoV-2 3CL protease activity by 3 compounds was evaluated using the fluorescence resonance energy transfer method (each compound was tested in triplicate).

Weighing appropriate amount of the above compounds, preparing a solution with appropriate concentration gradient with DMSO, mixing 5 μ L of the prepared solution with 91 μ L of Assay Reagent (Assay Buffer: SARS-CoV-2M) ^pro /3CL ^pro =90:1, purchased from shanghai bi yunnan biotechnology limited) were sequentially added to black 96-well plates, mixed, incubated at 37 ℃ for 10 minutes in the dark, and 4 μ L of Substrate (100 μ M Dabcyl-KTSAVLQSGFRKME-Edans, purchased from shanghai bi yunnan biotechnology limited) was rapidly added to each well, mixed. After incubation for 5min at 37 ℃ in the dark, the signal tends to be stable, fluorescence measurement is carried out within 5-30min by using a multifunctional microplate reader (Serimei Feishell science and technology Co., ltd., varioskan Flash) and the inhibition percentage of the sample is calculated, wherein the excitation wavelength is 340nm and the emission wavelength is 490nm. Assay Reagent without compound was used as 100% enzyme activity control, without SARS-CoV-2M ^pro The Assay Buffer of the/3 CL protease is a blank control, and the rest of the treatment methods are the same. Non-linear regression analysis using GraphPad Prism software to calculate the IC of the samples 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde, 1,4-dimethoxy-2-naphthaldehyde ₅₀ The value is obtained.

As shown in Table 2, examples 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde, 1,4-dimethoxy-2-naphthaldehyde all have inhibition effect on SARS-CoV-2 3CL proteaseThe activity of 2,3-dimethoxy-1-naphthaldehyde is stronger in inhibiting effect on SARS-CoV-2 3CL protease, and IC is ₅₀ The values are all around 20 mu M, and the inhibition effect of 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde and 1,4-dimethoxy-2-naphthaldehyde on SARS-CoV-2 CL protease is respectively shown in FIGS. 4-6.

TABLE 2 inhibition ratio of SARS-CoV-2 3CL protease (%)

In conclusion, 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde and 1,4-dimethoxy-2-naphthaldehyde show excellent inhibitory activity to 3CL protease of SARS-CoV and SARS-CoV-2 coronavirus, and can be used as an anti-coronavirus medicine for development and research.

Claims (10)

1. Application of naphthyl skeleton compound in preparing coronavirus 3CL protease inhibitor.

2. Application of naphthyl skeleton compound in preparing medicine for preventing and/or treating coronavirus caused diseases.

3. Use according to claim 1 or 2, characterized in that the naphthyl skeleton compound is a pharmaceutically acceptable salt, solvate, enantiomer, diastereomer, tautomer or mixture thereof in any proportion.

4. Use according to claim 1 or 2, characterized in that the naphthyl skeleton-like compound is one of the following compounds:

5. use according to claim 1 or 2, characterized in that the naphthyl skeleton like compound is 2,3-dimethoxy-1-naphthaldehyde, 2-hydroxy-7-methoxy-1-naphthaldehyde or 1,4-dimethoxy-2-naphthaldehyde.

6. Use according to claim 1 or 2, wherein the coronavirus is MERS-CoV, SARS-CoV or SARS-CoV-2.

7. The use according to claim 2, wherein the coronavirus-caused disease is an infectious disease caused by MERS-CoV, SARS-CoV-2 or a complication thereof.

8. The use according to claim 1 or 2, wherein the inhibitor or drug is in the form of tablets, capsules, granules, syrups, premixes or pellets for oral administration, or in the form of liniments or injections for non-oral administration.

9. The use of claim 1, wherein the inhibitor is a naphthyl backbone alone or in combination with other coronavirus 3CL protease inhibitors.

10. The use according to claim 2, wherein the medicament is a naphthyl scaffold alone or in combination with other medicaments for the prevention and/or treatment of diseases caused by coronaviruses.

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