CN113101283A

CN113101283A - Application of polyphenyl ring conjugated molecule in preparation of antiviral drug and antiviral drug

Info

Publication number: CN113101283A
Application number: CN202110405894.2A
Authority: CN
Inventors: 蒋兴宇; 贾跃晓
Original assignee: Southern University of Science and Technology
Current assignee: Southern University of Science and Technology
Priority date: 2021-04-15
Filing date: 2021-04-15
Publication date: 2021-07-13
Anticipated expiration: 2041-04-15
Also published as: CN113101283B

Abstract

The invention provides application of a polyphenyl ring conjugated molecule in preparation of an antiviral drug and the antiviral drug. The multi-benzene ring conjugated molecule has a core unit and a conjugated arm structure connected with the core unit, and contains at least two benzene rings. Dissolving the polyphenyl ring conjugated molecule in a dispersion medium, and quickly diluting to prepare the antiviral drug; the obtained antiviral drug has good antiviral effect, and can effectively inhibit and kill various viruses. The invention utilizes the polyphenyl ring conjugated molecule to prepare the medicine with high-efficiency antiviral effect, and provides an effective treatment scheme for treating in-vivo infection diseases caused by drug-resistant viruses and novel viruses.

Description

Application of polyphenyl ring conjugated molecule in preparation of antiviral drug and antiviral drug

Technical Field

The invention belongs to the technical field of antivirus, and particularly relates to application of a polyphenyl ring conjugated molecule in preparation of an antivirus medicament and the antivirus medicament.

Background

Viruses (viruses) are non-cellular, parasitically living organisms consisting of a single nucleic acid molecule (DNA or RNA) and a protein. Infectious diseases caused by viruses have the characteristics of strong infectivity, wide prevalence, high mortality and the like, and currently, the novel coronavirus (COVID-19) pneumonia has great abuse, thereby seriously threatening human life health and global public health safety. In addition, respiratory diseases caused by influenza virus and coronavirus, AIDS caused by HIV virus, hepatitis B caused by hepatitis B virus, hand-foot-and-mouth disease caused by enterovirus and the like have serious influence on the life health of human beings.

At present, the clinical use and the antiviral drugs reported in the prior art mainly comprise the following drugs:

(1) chemical drugs, in addition to common chemical antiviral drugs such as nucleosides, non-nucleosides, biological and polysaccharides, CN103768073A discloses the use of a steroid derivative with a dihydrazone structure as an antiviral drug, and also provides the use of a pharmaceutically acceptable salt of the steroid derivative with a dihydrazone structure as an antiviral drug, and an antiviral drug preparation consisting of an effective amount of the steroid derivative with a dihydrazone structure and acceptable auxiliary materials;

(2) chinese herbal medicines such as radix isatidis, honeysuckle, copperleaf antipyretic capsules and the like, for example, CN105168374A discloses an application of a fennel fruit extract in preparing antiviral drugs. The hypecoum leptocarpum extract is an extract which is obtained by extracting dry whole herbs of hypecoum leptocarpum of the genus hypecoum of the family papaveraceae with alcohol and purifying with macroporous resin column, has a total alkaloid content of 30-95% and a total flavone content of 5-60%, and at least comprises the hypecoum leptocarpum extract in various dosage forms of antiviral drugs which are composed of chemical drugs, traditional Chinese medicines or natural medicines. In vitro and in vivo experiments prove that the hypecoum leptocarpum extract can be used for preparing antiviral drugs;

(3) new antiviral drugs such as nano antiviral drugs, polymers, liposomes, etc.; for example, CN101288646A discloses a lipid nanoparticle with anti-hepatitis b virus activity, which is composed of an antiviral drug and a lipid material, wherein the drug is adefovir dipivoxil. The solid lipid nanoparticles and the nanostructure lipid carrier have rapid cell uptake and cytoplasm retention functions, and the molecular target is encapsulated with antiviral drugs in cells, so that the drug uptake of virus cells and the drug concentration of the drug molecular target part can be greatly increased. The intake of virus cells to the medicine is increased, which is beneficial to reducing the distribution of the medicine in normal tissues or cells and reducing the toxic and side effects of the medicine; the increase of the drug concentration of the drug molecule target part is beneficial to improving the curative effect of the antiviral drug, and can be applied to the preparation of the anti-hepatitis B virus drug.

However, due to the long-term use of the existing antiviral chemical drugs, most viruses generate drug resistance to the drugs, so that the curative effect of the viruses is reduced; antiviral Chinese herbal medicines have poor specificity to viruses and are often required to be combined with chemical medicines to achieve antiviral curative effect; the stability, toxicity and in vivo drug metabolism of novel antiviral drugs remain to be further studied. Meanwhile, new outbreaks of viral infectious diseases often require new antiviral drugs for treatment, and therefore, the development of new antiviral drugs is urgently needed.

Disclosure of Invention

In view of the problems in the prior art, the invention provides the application of the polyphenyl conjugated molecule in preparing the antiviral drug and the antiviral drug.

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

in a first aspect, the present invention provides a use of a polyphenyl cyclic conjugate molecule in the preparation of an antiviral medicament, the polyphenyl cyclic conjugate molecule having a core unit and a conjugate arm structure connected to the core unit; the multi-benzene ring conjugated molecule contains at least two benzene rings.

The polyphenyl ring conjugated molecule is used as an antiviral molecule, has a polyphenyl ring conjugated structure, and can be divided into a core structure and an arm structure, wherein the core structure can be any atom or group, and the arm structure with conjugated property is connected with the core. Such molecules may act as antiviral by binding to the viral 3CL protease. All coronaviruses encode a 3CL protease, which is used for post-translational processing of proteins. Therefore, the inhibition of the function of the 3CL protease can effectively block the reproduction of viruses, thereby playing an antiviral role.

The invention utilizes the polyphenyl ring conjugated molecule to prepare the medicine with high-efficiency antiviral effect, is used for resisting the virus resistant to the traditional medicine and the new outbreak of virus infection, and provides an effective scheme for treating in-vivo infectious diseases caused by drug resistant virus and novel virus.

As a preferable technical scheme of the invention, the number of the conjugated arm structures is 2-6.

Preferably, the conjugated arm structures include, but are not limited to: alkane chains, benzoic acid, diphenic acid,

Or a combination of any one or at least two of carboxyphenylethynyl; wherein,

indicates the position of the group's attachment.

Preferably, the core unit includes, but is not limited to: carbon atom, nitrogen atom, sulfur atom, silicon atom, vinyl group, tetraphenyl ethylene, benzene ring or 1,3, 5-triazine,

Any one or a combination of at least two of them, wherein,

indicates the position of the group's attachment.

Specifically, the multi-benzene ring conjugated molecule is any one of the following 1-46 molecules:

as a preferable technical scheme of the invention, the concentration of the poly-benzene ring conjugated molecule is more than or equal to 0.01 mu g/mL. In the invention, the polyphenyl ring conjugated molecule has better antiviral ability, and can still exert better antiviral ability under lower concentration such as 0.01 mu g/mL; therefore, the concentration of the poly (benzene ring) -conjugated molecule of the present invention may be 0.01. mu.g/mL, 0.05. mu.g/mL, 0.1. mu.g/mL, 0.2. mu.g/mL, 0.5. mu.g/mL, 1. mu.g/mL, 2. mu.g/mL, 10. mu.g/mL, 100. mu.g/mL, 500. mu.g/mL, 1000. mu.g/mL, 5000. mu.g/mL, 10000. mu.g/mL, or the like.

Preferably, the polyphenyl ring conjugated molecule is used against DNA viruses and/or RNA viruses.

Preferably, the multi-phenyl ring conjugate molecule is used against any one of or a combination of at least two of a novel coronavirus, influenza virus or Respiratory Syncytial Virus (RSV).

In a second aspect, the present invention provides an antiviral drug comprising at least one polyphenylenic conjugated molecule and a dispersion medium.

The multi-benzene ring conjugated molecule comprises a core unit and a conjugated arm structure connected with the core unit, and has at least two benzene rings.

Preferably, the antiviral drug comprises any one or a combination of at least two of the polyphenyl ring conjugated molecules 1-46.

As a preferable embodiment of the present invention, the dispersion medium includes a liquid dispersion medium or a solid dispersion medium.

Preferably, the liquid dispersion medium includes any one of N, N-dimethylformamide, ethyl acetate, hexane, tetrahydrofuran, N-dimethylformamide, ethyl acetate, hexane, dimethylsulfoxide, a liquid medium, a phosphate buffer, serum, a glucose injection solution, or a physiological saline, or a combination of at least two thereof;

in the present invention, when the polyphenyl ring conjugate molecule is used to prepare an antiviral drug, the polyphenyl ring conjugate molecule can be dissolved by any organic solvent (including but not limited to tetrahydrofuran, N-dimethylformamide, ethyl acetate, hexane, etc.), and can be dissolved to any concentration less than 20 mg/mL;

thereafter, it may be diluted with any aqueous solution (including but not limited to, e.g., purified water, physiological saline, phosphate buffered saline, cell culture media, bacterial culture media, etc.) or organic solution (e.g., tetrahydrofuran, N-dimethylformamide, ethyl acetate, hexane, etc.) to any concentration for antiviral therapy.

Preferably, the antiviral drug further comprises pharmaceutically acceptable excipients.

Preferably, the dosage form of the antiviral drug comprises any one or the combination of at least two of common compressed tablets, dispersible tablets, enteric-coated tablets, capsules, granules, dripping pills, emulsions, powder, oral liquid or injections.

Preferably, the administration route of the antiviral drug includes 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 them.

In the invention, the antiviral drug prepared by the polyphenyl ring conjugated molecule can be used for treating any virus infection diseases, including but not limited to virus infections such as skin infection, abdominal cavity infection, lung infection, meningitis, intestinal infection and the like; but also for any form of sterilisation and disinfection treatment. The antiviral drug can also be made into any form of antiviral product, apparatus, etc., for preventing and treating viral infection.

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 at least the following beneficial effects:

(1) the polyphenyl ring conjugated molecule has a polyphenyl ring conjugated structure, the structure of the polyphenyl ring conjugated molecule can be divided into a core and an arm structure, the core can be any atom or group, and the arm structure with conjugated property is connected with the core; the preparation methods of the polyphenyl ring conjugated molecules can be obtained from the prior art, and are convenient to prepare;

(2) in the invention, the multi-benzene ring conjugated molecule is dissolved in organic solution such as dimethyl sulfoxide and the like, and the antiviral medicament can be prepared by fast dilution; the obtained antiviral drug has good antiviral effect, and can effectively inhibit and kill various viruses;

taking poly-benzene ring conjugated molecule 6 as an example, the EC thereof₅₀As low as 1. mu.g/mL, and TD₅₀Is 64 mug/mL, has good treatment effect in an in-vivo virus infection model, and has the cure rate of 100 percent;

(3) the invention utilizes the polyphenyl ring conjugated molecule to prepare the medicine with high-efficiency antiviral effect, is used for resisting the virus resistant to the traditional medicine and the new outbreak of virus infection, and provides an effective scheme for treating in-vivo infectious diseases caused by drug resistant virus and novel virus.

Drawings

FIG. 1 is a scattergram obtained by examining the antiviral activity and cytotoxicity of the multi-benzene ring conjugate molecule 6 in example 2.

FIG. 2 is a bar graph of the inhibitory effect of poly-benzene ring conjugated molecules 1-10 on GX _ P2V virus.

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, reagents and instruments used are available from conventional sources unless otherwise specified, and the experimental procedures used are those known to those skilled in the art.

Example 1

This embodiment takes the poly-benzene ring conjugated molecule 6 as an example, and provides an application of the poly-benzene ring conjugated molecule in preparing antiviral drugs.

(1) Preparation of poly-benzene ring conjugated molecule 6

The synthetic route of the poly-benzene ring conjugated molecule 6 can be synthesized according to any method disclosed in the art, and in this example, the synthetic route of the molecule 6 is as follows:

(2) preparation of antiviral drug based on multi-benzene ring conjugated molecule 6

Adding the poly-benzene ring conjugated molecule 6 into DMSO, and performing ultrasonic treatment to fully dissolve the molecule, wherein the concentration of the molecule 6 in the DMSO solution of the poly-benzene ring conjugated molecule 6 is 4096 mug/mL for later use;

the DMSO solution of the polyphenyl ring conjugated molecule 6 can be directly added into a culture medium, serum, glucose injection or normal saline and other solvents for in vivo and in vitro antiviral research;

in this example, an antiviral drug was prepared by diluting a DMSO solution containing molecule 6 to 256. mu.g/mL with ultrapure water.

Example 2

This example uses a cell culture method to determine the activity of antiviral drugs.

MDCK cells were cultured and infected with a 2019-nCoV similar virus GX-P2V and observed for cytopathic effect (CPE) upon addition of the antiviral drug.

The method comprises the following specific steps:

MDCK cells were cultured in 96-well plates, and after removing the medium, a virus culture solution (10) was added thereto⁶CFU/mL); then, antiviral drugs 6 were added to each well at different concentrations (0.1. mu.g/mL, 1. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL, 20. mu.g/mL, 50. mu.g/mL, 100. mu.g/mL and 200. mu.g/mL), and they were cultured in an incubator at 37 ℃ for 3 days;

observing cytopathic effect, and calculating half toxic concentration TD of the test drug according to Reed-Muench method₅₀And half the effective concentration EC₅₀And calculating the Therapeutic Index (TI) of the antiviral drug.

The antiviral activity and cytotoxicity of the polyphenyl-ring conjugated molecule 6 are shown in figure 1, and the EC thereof is₅₀As low as 1. mu.g/mL, and TD₅₀It is 64. mu.g/mL, and its therapeutic index TI is 64.

Example 3

This example was used to study the therapeutic effect of antiviral drugs in respiratory infections in mice. The specific implementation process is as follows:

15 mice were divided into two groups, one group consisting of 10 mice in the experimental group and one group consisting of 5 mice in the control group.

Into the nasal cavity of 15 miceSlowly drop into 10⁹100 μ L of GX _ P2V virus suspension in CFU/mL; injecting 100 μ L antibacterial agent (with dosage of 1, 2, 4, 8, 16mg/kg) into 10 mice of the experimental group via tail vein injection after 30min, and injecting two mice per dosage;

observing the survival rate, the body weight, the fur and the dehydration state of the mice of the experimental group and the control group after 3 days, taking the lungs of the mice, observing the pathological changes and the inflammatory reaction of the lungs through tissue sections, and measuring the virus content of the lungs.

Through culture and determination, the survival rate of the mice in the control group is 0%, and the survival rate of the mice in the experimental group injected with the benzene ring conjugated molecule 6 is 100%.

The experimental results prove that the antiviral drug provided by the embodiment can effectively treat respiratory tract infection caused by virus, and the polyphenyl ring conjugated molecule 6 with the concentration of 2mg/kg can effectively relieve infection symptoms of mice, improve lung infection and eliminate lung virus.

Example 4

In this example, polyphenyl ring conjugated molecules 1-5 and 7-10 were prepared according to example 1 and conventional methods in the art, and the activity of antiviral drugs was measured by cell culture.

MDCK cells were cultured in a 96-well plate, and after removing the medium, a GX-P2V virus culture medium (10) was added thereto⁶CFU/mL); then adding 1-5 and 7-10 mu g/mL of antiviral drugs into each hole respectively, and culturing the mixture in an incubator at 37 ℃ for 3 days;

the antiviral activities of the polyphenyl ring conjugated molecules 1-5, 7-10 and the polyphenyl ring conjugated molecule 6 related in the embodiment 2 are shown in figure 2;

as can be seen from the figure, compared with a control group, the antiviral drugs prepared from the polyphenyl ring conjugated molecules 1-10 have better inhibitory effect on GX-P2V virus.

In addition, the same method is also used for detecting the inhibition effect of the polyphenyl ring conjugated molecules 1-10 on different viruses, such as influenza virus IFV (FM1 strain) and respiratory syncytial virus RSV (Long strain), and the experimental result proves that the polyphenyl ring conjugated molecules can also play a better virus inhibition effect; the invention also detects the antiviral effect of the polyphenyl ring conjugated molecules 11-46, and the obtained polyphenyl ring conjugated molecules 11-46 have the antiviral effect.

In conclusion, the invention provides the application of the polyphenyl ring conjugated molecule in preparing the antiviral drug and the antiviral drug, and provides an effective treatment scheme for treating in vivo infection diseases caused by drug-resistant viruses and novel viruses.

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.

Claims

1. The application of the multi-benzene ring conjugated molecule in preparing the antiviral drug is characterized in that the multi-benzene ring conjugated molecule has a core unit and a conjugated arm structure connected with the core unit;

the multi-benzene ring conjugated molecule contains at least two benzene rings.

2. Use according to claim 1, wherein the number of conjugated arm structures is 2-6;

preferably, the conjugated arm structure comprises an alkane chain, benzoic acid, diphenic acid, methyl ethyl ketone, ethyl methyl ketone,

Or a combination of at least two of carboxyphenylethynyl, wherein,

represents an access position of a group;

preferably, the core unit includes a carbon atom, a nitrogen atom, a sulfur atom, a silicon atom, a vinyl group, a tetraphenylethylene, a benzene ring, a 1,3, 5-triazine, a,

Any one or a combination of at least two of them, wherein,

indicates the position of the group's attachment.

3. Use according to claim 1 or 2, characterized in that the polycyclic conjugated molecule is one of the following molecules:

4. use according to any one of claims 1 to 3, wherein the concentration of the poly-phenyl ring conjugated molecule used is not less than 0.01 μ g/mL.

5. The use according to any one of claims 1 to 4, wherein the polyphenyl ring conjugated molecule is used against DNA viruses and/or RNA viruses;

preferably, the multi-phenyl ring conjugated molecule is used against any one of or a combination of at least two of a novel coronavirus, influenza virus or respiratory syncytial virus.

6. An antiviral drug comprising at least one polyphenyle conjugate molecule and a dispersing medium;

7. The antiviral drug of claim 6, wherein said conjugated arm structure comprises an alkane chain, benzoic acid, bibenzoic acid,

Or a combination of any one or at least two of carboxyphenylethynyl;

preferably, the number of the conjugated arm structures is 2-6;

preferably, the core unit comprises a carbon atom, a nitrogen atom, a sulfur atom, a silicon atom, a vinyl group, a tetraphenylethylene, a benzene ring or a 1,3, 5-triazine,

Any one or a combination of at least two of them, wherein,

indicates the position of the group's attachment.

8. The antiviral drug according to claim 6 or 7, wherein the polyphenyl ring conjugated molecule is selected from the following molecules:

9. the antiviral agent according to any one of claims 6 to 8, wherein the dispersion medium comprises a liquid dispersion medium or a solid dispersion medium;

10. The antiviral drug according to any one of claims 6 to 9, wherein the dosage form of the antiviral drug comprises any one or a combination of at least two of ordinary compressed tablets, dispersible tablets, enteric-coated tablets, capsules, granules, dropping pills, emulsions, powders, oral liquids or injections;