CN114099600A

CN114099600A - Application of ciliate desert-grass in preventing and treating virus infection

Info

Publication number: CN114099600A
Application number: CN202111392089.7A
Authority: CN
Inventors: 聂棱; 梁友东
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-11-23
Filing date: 2021-11-23
Publication date: 2022-03-01

Abstract

The invention provides a composition for preventing and/or treating virus infection, which comprises shortleaf kyllinga herb or extract thereof; wherein the virus comprises an RNA virus and a DNA virus; the RNA virus is an RNA virus causing fever infectious diseases and is selected from one or more of HIV virus, rabies virus, Ebola virus, Marburg virus, Lassa virus, hepatitis A virus and hand-foot-and-mouth virus, and influenza virus and herpes virus are not included; the DNA virus is selected from one or more of hepatitis B virus and HPV virus. Particularly, the invention discovers that the shortleaf kyllinga herb can effectively inhibit HIV virus, so that the shortleaf kyllinga herb can effectively prevent and/or treat virus infection or diseases caused by the virus infection and can be applied as an antiviral medicament.

Description

Application of ciliate desert-grass in preventing and treating virus infection

Technical Field

The invention belongs to the technical field of pharmacy and antivirus, and particularly relates to application of shortleaf kyllinga herb in preventing and/or treating virus infection.

Background

A virus is a noncellular organism that is small, simple in structure, must be parasitic in living cells and proliferated in a replicative manner. The chemical components of the viral nucleic acid are DNA or RNA, and the viruses are divided into two main categories, namely DNA viruses and RNA viruses. Nucleic acids can be linear or circular and can be divided into single or double strands.

DNA viruses are widely present in humans, vertebrates, insects, and in a variety of continuous cell lines, each virus infects only one animal (with individual exceptions) and causes only a few diseases.

RNA viruses have two replication modes, namely self-replication and reverse transcription, and in the replication process of viral RNA, the activity of enzyme of an error repair mechanism is very low and almost none, so that the mutation is very quick. RNA viruses are more susceptible to disease, are more lethal to the host, are more mutable, and are more diverse than DNA viruses, are more difficult to develop effective vaccines, and are difficult to prevent.

HIV, a Human Immunodeficiency Virus, is a Virus that causes a defect in the Human immune system. The virus destroys the immunity of the human body, resulting in the loss of resistance of the immune system, thereby causing various diseases and cancers to be preserved in the human body, and finally resulting in Acquired Immune Deficiency Syndrome (AIDS).

The HIV virus is a Lentivirus (Lentivirus) that infects cells of the human immune system, and belongs to a kind of retrovirus. AIDS virus mainly invades the immune system of human body, including CD4+ T lymphocyte, monocyte macrophage and DC, etc., and is mainly characterized in that the quantity of CD4+ T lymphocyte is continuously reduced, finally the cell immune function of human body is deficient, and various opportunistic infections and tumors are caused. Aids virus infection increases the risk of cold, influenza and pneumonia, as well as the risk of pulmonary hypertension. In addition, HIV also makes it easier for infected individuals to infect tuberculosis.

At present, the modern medicine for treating AIDS mainly comprises four methods: antiviral therapy, immunodeficiency therapy, treatment of symptomatology infection and tumor therapy, but medicines commonly used in western medicine may have large side effects, such as liver and kidney injury, rash, granulocytopenia and the like.

The traditional medicine treatment has obvious advantages in preventing diseases, improving clinical symptoms, preventing mild to severe and severe to critical.

The shortleaf kyllinga herb is a cyperaceae plant shortleaf kyllinga herb, the rhizome of which grows and creeps, and has a plurality of internodes, the length of the internodes is about 1.5 cm, and each node is provided with a stalk. The chemical components of the shortleaf kyllinga herb contain volatile oil, and the shortleaf kyllinga herb can be used for treating cold and wind-cold, fever and coldness headache, arthralgia and myalgia, cough, malaria, jaundice, dysentery, pyocutaneous disease and pyogenic infections, and traumatic injuries and knife wounds. At present, the application of the shortleaf meadowsweet in the aspect of antivirus is not much concerned.

Disclosure of Invention

The invention aims to provide a composition for preventing and/or treating virus infection and application thereof in resisting virus.

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

in a first aspect, the invention provides the use of ciliate desert-grass or an extract thereof for the manufacture of a medicament and/or food for the prevention and/or treatment of viral infections or diseases caused by viral infections.

The shortleaf kyllinga herb is a centipede belonging to the family Cyperaceae.

The Grateloupia filicina can be applied in the form of hay, powder or extract. The shortleaf kyllinga herb extract refers to an aqueous extract or an organic solvent extract of shortleaf kyllinga herb, wherein the organic solvent is selected from one or more of methanol, ethanol, propanol, acetone, petroleum ether, diethyl ether and ethyl acetate, and the preferable solvent is selected from one or more of water, ethanol, diethyl ether and ethyl acetate.

The virus comprises RNA virus and DNA virus, the RNA virus is RNA virus causing fever infectious disease, and is selected from one or more of HIV virus, Ebola virus, Marburg virus, Lassa fever virus, rabies virus, hepatitis A virus and hand-foot-and-mouth virus, with the proviso that influenza virus and herpes virus are not included; the DNA virus is selected from one or more of hepatitis B virus and HPV virus.

Preferably, the virus is selected from one or more of HIV virus, ebola virus, rabies virus, HPV virus and hepatitis b virus.

The diseases include AIDS, hemorrhagic fever, rabies, hepatitis A, hand, foot and mouth, respiratory tract infection, pharyngolaryngitis, rash, lymphadenectasis, hepatitis B and/or cervical infection. Preferably, the disease is acquired immunodeficiency syndrome and its complications caused by HIV infection.

In a second aspect, the present invention provides the use of ciliate desert-grass or an extract thereof for the prophylactic and/or therapeutic treatment of a viral infection or a disease caused by a viral infection.

The shortleaf kyllinga herb is a centipede belonging to the family Cyperaceae.

In a third aspect, the present invention provides a composition for preventing and/or treating viral infection, comprising Grateloupia filicina or an extract thereof.

The shortleaf kyllinga herb is a centipede belonging to the family Cyperaceae.

In some embodiments, the virus is an HIV virus.

In some embodiments, the virus is an ebola virus.

In some embodiments, the virus is a rabies virus.

In some embodiments, the virus is an HBV virus.

In some embodiments, the virus is an HPV virus.

Preferably, the composition is a pharmaceutical composition, which further comprises a pharmaceutically acceptable adjuvant and/or carrier.

Preferably, the pharmaceutical composition is in a form of gastrointestinal administration, injection administration or respiratory administration.

Preferably, the pharmaceutical composition is in a dosage form selected from the group consisting of powder, tablet, granule, capsule, solution, emulsion, suspension, injection, spray, aerosol, and powder spray.

Preferably, the composition is a food composition, which further comprises a dietetically acceptable adjuvant and/or carrier.

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

(1) the invention discovers that the shortleaf meadowrue herb can effectively inhibit cell infection caused by HIV pseudovirus, and therefore, the shortleaf meadowrue herb can be used for preventing and/or treating diseases caused by HIV virus infection.

(2) The shortleaf kyllinga herb or the extract thereof or the composition containing the shortleaf kyllinga herb provided by the invention can effectively prevent and/or treat diseases (including AIDS, hemorrhagic fever, rabies, hepatitis A, hand, foot and mouth, respiratory tract infection, sphagitis, rash, lymphadenectasis, hepatitis B and/or cervical infection and the like) caused by virus infection, and can be applied as an antiviral medicament.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.

Example 1

This example provides an aqueous extract of Grateloupia filicina.

8.9 g of dry centipede herb, 500 ml of water is added after the dry centipede herb is cleaned, the mixture is heated for 20 minutes, and supernatant is taken and filtered for standby.

Example 2

This example provides a 70% ethanol extract of Grateloupia filicina.

8.9 g of shortleaf meadowrue herb, after being cleaned, 500 mL of 70% ethanol is added, the mixture is heated for 20 minutes, the supernatant is taken out, filtered and evaporated to dryness, and 450mL of water is added to dissolve the supernatant for standby.

Example 3

This example provides a composition comprising an ether extract of Grateloupia filicina and a cyclodextrin adjuvant.

8.9 g of ciliate desert-grass, after being cleaned, 500 mL of ether is added, the mixture is heated for 20 minutes, the supernatant is taken out, filtered and evaporated to dryness, 450mL of water is added for dissolution, and a proper amount of cyclodextrin is added for standby.

Experimental example 1 Performance test

The substances provided in examples 1 to 3 were subjected to a performance test, and whether the sample had an inhibitory effect on the HIV pseudovirus was judged by the following test method:

a. dilution of

The sample solution was diluted with the complete medium in 3-fold, 9-fold, 27-fold, 81-fold, 243-fold, 729-fold, 2187-fold, 6561-fold, and 19683-fold.

b. Experiment of

The HIV neutralization test comprises two parts, namely pseudovirus preparation and detection after pseudovirus infection, wherein the pseudovirus preparation is realized by cotransfecting eukaryotic expression cells (293FT) by two plasmids: one of the plasmids is pSG3 Δ Env, which expresses all genes of HIV except Env (i.e. backbone plasmid); another plasmid is pcDNA3.1-Env, which expresses the full-length Env gene. In pseudovirus production, only the backbone plasmid is transcribed into viral genomic RNA and packaged into pseudovirus, and thus the pseudovirus produced does not have the replication ability to produce progeny virus, but only has a single round of infection. TZM-bl cells are HeLa cells with modified CXCR4+, the modified cells stably express surface molecules CD4 and CCR5, and further modification enables the genome of the cells to integrate firefly luciferase reporter genes, and the expression of the firefly luciferase reporter genes is controlled by HIV long terminal repeat sequences. In the pseudovirus infection process, the HIV pseudovirus strain infects TZM-bl cells through a cell surface receptor and an auxiliary receptor, the intracellular Tat gene is expressed, the expressed Tat protein activates the expression of a reporter gene in cis, and after a corresponding luciferase substrate is added, a generated luminescence signal (relative luminescence unit, RLU) is in direct proportion to the number of viruses infecting the cells.

Incubation and sample application

Plate paving: TZM-bl cells were seeded in 96-well plates and cultured for 24 h.

Sample group: the diluted samples were mixed with HIV pseudovirus (200TCID50) in 50. mu.L aliquots, incubated for 1h at 37 ℃ in an incubator, and added to pre-plated TZM-bl cells.

Positive control group: HIV pseudovirus (200TCID50) and complete medium were mixed in equal volumes of 50. mu.L each, incubated for 1h at 37 ℃ in an incubator, and added to pre-plated TZM-bl cells.

Negative control group: mu.L of complete medium was taken, incubated for 1h at 37 ℃ in an incubator and added to the pre-plated TZM-bl cells.

In the sample group, the positive control group and the negative control group, after 1 hour of addition to the TZM-bl cells plated in advance, DEAE accelerating agent was added to a final concentration of 15. mu.g/ml at 37 ℃ with 5% CO₂The cultivation was continued in the incubator for 48 h. The chemiluminescence apparatus performs luminescence value detection on samples in a 96-well plate.

Inhibition (%) < 1- (mean sample RLU-mean negative control RLU)/(mean positive control RLU-mean negative control RLU).

When the inhibition rate is less than 50%, it indicates that the sample has substantially no neutralization inhibition.

Table 1 shows the results of the testing of the materials provided in example 1:

TABLE 1

Sample dilution/fold	RLU	Inhibition rate/%)
			Stock solution	4.74E+04	97.42
3	7.26E+04	96.04
			9	1.60E+05	91.24
27	3.76E+05	79.44
			81	8.68E+05	52.51
243	1.22E+06	33.22
			729	1.28E+06	29.73
2187	1.50E+06	17.50
			6561	1.58E+06	13.37
19683	1.69E+06	7.53
			Positive control	1.83E+06	-

Note: the RLU value for the negative control was 270.

Table 2 shows the results of testing the substances provided in examples 1-3 (3-fold dilution):

TABLE 2

Sample (I)	RLU	Inhibition rate/%)
			Example 1	7.26E+04	96.04
Example 2	1.58E+05	91.37
			Example 3	2.24E+05	87.76

As can be seen from the examples and performance tests, the substance or the composition provided by the invention has a good inhibiting effect on the HIV pseudovirus, and can be used for preventing and/or treating diseases or symptoms caused by HIV virus infection.

The applicant states that the present invention is illustrated by the above examples of compositions for the prevention and/or treatment of viral infections according to the present invention, but the present invention is not limited to the above detailed methods, i.e. it is not meant to imply that the present invention must rely on the above detailed methods for its implementation. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims

1. Use of ciliate desert-grass or its extract for the preparation of a medicament and/or food for the prevention and/or treatment of viral infections or diseases caused by viral infections, wherein the viruses include RNA viruses and DNA viruses, the RNA viruses are RNA viruses causing febrile infectious diseases, selected from one or more of HIV viruses, ebola viruses, marburg viruses, lassa fever viruses, rabies viruses, hepatitis a viruses, hand-foot-and-mouth viruses, with the proviso that influenza viruses and herpes viruses are excluded; the DNA virus is selected from one or more of hepatitis B virus and HPV virus.

2. The use according to claim 1, wherein the virus is selected from one or more of the group consisting of HIV virus, ebola virus, rabies virus, HPV virus and hepatitis b virus.

3. The use of claim 1, wherein the disease comprises aids, hemorrhagic fever, rabies, hepatitis a, hand, foot and mouth, respiratory tract infections, pharyngolaryngitis, rash, lymphadenectasis, hepatitis b and/or cervical infections.

4. Use of ciliate desert-grass or an extract thereof for the prevention and/or treatment of a viral infection or a disease caused by a viral infection, wherein the virus comprises an RNA virus and a DNA virus, the RNA virus is an RNA virus causing febrile infectious disease and is selected from one or more of HIV virus, ebola virus, marburg virus, lassa fever virus, rabies virus, hepatitis a virus, hand-foot-and-mouth virus, with the proviso that influenza virus and herpes virus are excluded; the DNA virus is selected from one or more of hepatitis B virus and HPV virus.

5. The use according to claim 4, wherein the virus is selected from one or more of the group consisting of HIV virus, Ebola virus, rabies virus, HPV virus and hepatitis B virus.

6. The use of claim 4, wherein the disease is selected from the group consisting of AIDS, hemorrhagic fever, rabies, hepatitis A, hand, foot and mouth, respiratory tract infections, pharyngolaryngitis, rash, lymphadenectasis, hepatitis B and/or cervical infections.

7. A composition for preventing and/or treating viral infection, comprising Grateloupia filicina or its extract;

wherein the virus comprises RNA virus and DNA virus, the RNA virus is RNA virus causing fever infectious disease, and is selected from one or more of HIV virus, Ebola virus, Marburg virus, Lassa fever virus, rabies virus, hepatitis A virus and hand-foot-and-mouth virus, with the proviso that influenza virus and herpes virus are not included; the DNA virus is selected from one or more of hepatitis B virus and HPV virus;

8. The composition of claim 7, wherein the composition is a pharmaceutical composition, further comprising a pharmaceutically acceptable excipient and/or carrier.

9. The composition of claim 8, wherein the pharmaceutical composition is in a form for gastrointestinal administration, injection administration or respiratory administration;

10. The composition according to claim 7, wherein the composition is a food composition, further comprising a dietetically acceptable adjuvant and/or carrier.