CN106729635A - A kind of applications of IFN λ 3 in prevention or treatment AIDS-treating medicine is prepared - Google Patents

Abstract

The present invention relates to a kind of applications of IFN λ 3 in prevention or treatment AIDS-treating medicine is prepared, belong to biological pharmacy technical field.Medicine prepared by IFN λ 3 of the invention can effectively suppress the activity of HIV infection human macrophage, induce the expression of antiviral gene, good to AIDS virus inhibition.

Description

Application of a kind of IFN-λ3 in the preparation of drugs for preventing or treating AIDS

technical field

The invention relates to the technical field of biopharmaceuticals, in particular to an application of IFN-λ3 in the preparation of drugs for preventing or treating AIDS.

Background technique

At present, AIDS is still a global public health problem, the main reason being that there is still no specific medicine to completely cure AIDS and a vaccine that can prevent HIV infection.

Currently, the most commonly used AIDS treatment method in clinical practice is a combination drug method, and the most commonly used first-line treatment drugs are tenofovir (TDF) or zidovudine (AZT) + lamivudine (3TC) + efavirenz ( EFV) or nevirapine (NVP), commonly known as cocktail therapy, although this method can significantly inhibit HIV in patients, it still cannot be completely cured; and it is easy to cause HIV mutation, drug resistance, and has greater side effects on patients.

Interferons, a group of active proteins with multiple functions, are cytokines produced by monocytes and lymphocytes. In addition to antiviral and immunoregulatory effects, it also has obvious anti-cell proliferation effects. According to the characteristics of interferon receptors on the cell surface, interferons can be divided into type I interferon, type II interferon and type III interferon. Type III interferon (IFN-λ) is a new type of interferon, including three Family members: IL28A (IFN-λ2), IL28B (IFN-λ3) and IL29 (IFN-λ1). It is reported in the prior art that type III interferon has the function of regulating NK cells and has a good effect on the treatment of hepatitis, but there is no relevant report about type III interferon-3 (IFN-λ3) in the prevention or treatment of AIDS.

Contents of the invention

The purpose of the present invention is to provide an application of IFN-λ3 in the preparation of drugs for preventing or treating AIDS. The medicine prepared by the IFN-λ3 of the invention can effectively inhibit the activity of HIV-infected human macrophages, induce the expression of antiviral genes, and has a good inhibitory effect on HIV.

The invention provides an application of IFN-λ3 in the preparation of drugs for preventing or treating AIDS.

Preferably, the IFN-λ3 acts on the JAK-STAT signaling pathway, regulates signaling molecules, produces broad-spectrum antiviral factors, and inhibits HIV.

Preferably, the broad-spectrum antiviral factors include ISG56, OAS-1, MxA, A3G and A3F.

Preferably, said IFN-λ3 inhibits the activity of macrophages in HIV-infected humans.

The present invention also provides a drug for preventing or treating AIDS prepared from IFN-λ3, which is characterized in that the purity of the IFN-λ3 is greater than 95%.

Preferably, the drug includes IFN-λ3 and acceptable auxiliary materials.

Preferably, the concentration of the IFN-λ3 is 12.5-100 ng/ml.

The invention provides the application of IFN-λ3 in the preparation of drugs for preventing or treating AIDS, and can provide a drug for preventing or treating AIDS with high efficiency. The obtained drug can act on the JAK-STAT signaling pathway, regulate signaling molecules, and produce various anti-AIDS drugs. Viral factor, suppresses HIV and thus has the activity of suppressing macrophages in HIV-infected humans. The drug has no side effects, and compared with the existing chemical AIDS treatment drugs, it will not cause gene mutation and drug resistance. The experimental results showed that the inhibitory effect of IFN-λ3 was significantly higher than that of IFN-λ2 and IFN-λ3.

Description of drawings

Fig. 1 is the analysis diagram of the effect of IFN-λ3 inhibiting HIV provided by Example 1 of the present invention;

Fig. 2 is a graph showing the comparison results of the HIV-inhibiting effects of IFN-λ3, IFN-λ1 and IFN-λ2 provided in Example 1 of the present invention.

detailed description

The invention provides an application of IFN-λ3 in the preparation of drugs for preventing or treating AIDS.

In the present invention, the IFN-λ3 acts on the JAK-STAT signaling pathway, regulates signaling molecules, produces broad-spectrum antiviral factors, and inhibits HIV.

In the present invention, the broad-spectrum antiviral factors include ISG56, OAS-1, MxA, A3G and A3F.

In the present invention, the IFN-λ3 inhibits the activity of macrophages in HIV-infected humans.

The present invention also provides a drug for preventing or treating AIDS prepared from IFN-λ3, which is characterized in that the purity of the IFN-λ3 is greater than 95%. The present invention has no special limitation on the source of the IFN-λ3, and commercially available IFN-λ3 products well known to those skilled in the art can be used, such as IFN-λ3 sold by R&D Systems. In the present invention, there is no special limitation on the dosage form of the medicine, and the dosage form well known to those skilled in the art can be used.

In the present invention, the medicine includes IFN-λ3 and acceptable auxiliary materials.

In the present invention, the concentration of IFN-λ3 is 12.5-100 ng/ml, more preferably 100 ng/ml.

The application of the IFN-λ3 of the present invention in the preparation of drugs for preventing or treating AIDS will be further described in detail below in conjunction with specific examples. The technical solutions of the present invention include but are not limited to the following examples.

Example 1

Using an in vitro model of HIV-infected human macrophages, the activity of HIV reverse transcriptase was quantitatively detected by radioimmunoassay, and it was proved that IFN-λ3 has the activity of inhibiting HIV-infected human macrophages, which was detected by direct immunofluorescence technique Intracellular HIV p24 antigen also demonstrated this activity. The inhibitory effect is related to the dose and duration of IFN-λ3, that is, the larger the dose and the longer the duration, the better the inhibitory effect, and the effect before exposure is better than after exposure. Fig. 1 is an analysis diagram of the effect of IFN-λ3 inhibiting HIV. In Figure 1A, after 4 days of HIV infection, human macrophages were treated with 4 concentrations of IFN-λ3 for 8 days, and the HIV inhibitory effect was positively correlated with the concentration of IFN-λ3, among which 100ng/ml IFN-λ3 had the best inhibitory effect. Well, up to 80% or more. In Figure 1B, the human macrophages infected with HIV 4 days after treatment with 100ng/ml IFN-λ3, the inhibitory effect of IFN-λ3 gradually increased from 91.65% on the 8th day after infection to 96.6% on the 16th day, It shows that the effect of IFN-λ3 is positively correlated with the action time. In Figure 1C, before HIV infection, the macrophages were treated with IFN-λ3 (100ng/ml) 24 hours before HIV infection, and the infection also refers to the addition of IFN-λ3 (100ng/ml) and HIV to human macrophages at the same time In the cell culture vessel, post-infection refers to the addition of IFN-λ3 (100ng/ml) after 4 days of HIV infection. The inhibition effect reached 98.21%. Whereas if IFN-λ3 was withdrawn after 4 days of HIV infection, which would lead to HIV rebound, the inhibitory effect of IFN-λ3 at 12 days after HIV infection was only 68.2%.

Further research on the mechanism of action detected by fluorescent quantitative PCR method showed that IFN-λ3 mainly exerted the effect of inhibiting HIV through the JAK-STAT signaling pathway: compared with the control without IFN-λ3, IFN-λ3 could significantly up-regulate JAK-STAT The expression of Toll-like receptor molecules (TLR), interferon regulatory factor (IRF), and lambda interferon receptor molecules in the signaling pathway, and ultimately lead to intracellular interferon-stimulated gene 56 (ISG56), anti-myxovirus A High expression of a series of broad-spectrum antiviral factors such as (MxA), oligoadenylate synthase gene-1 (OAS-1), APOBEC3G (A3G), APOBEC3F (A3F), Tetherin, so as to achieve the effect of anti-HIV .

In the experiment of adding JAK inhibitors before adding IFN-λ3, it was found that the effect of IFN-λ3 in inhibiting HIV was significantly reduced, and the antiviral factors such as ISG56 and OAS-1 mediated by it were also significantly reduced. It further proves that IFN-λ3 regulates the signaling pathway molecules through the JAK-STAT signaling pathway, and produces a series of broad-spectrum antiviral factors to achieve the effect of inhibiting HIV.

Compared with IFN-λ1 and IFN-λ2, IFN-λ3 has a more obvious inhibitory effect, and its production of broad-spectrum antiviral factors such as ISG56 is also significantly higher than that of type 1 and type 2 λ interferon. The effect of IFN-λ3 on inhibiting HIV is compared with that of IFN-λ1 and IFN-λ2 as shown in Figure 2. In Figure 2A, 4 days after human macrophages were infected with HIV, the same dose (100ng/ml) of IFN-λ1, IFN-λ2, and IFN-λ3 were added respectively, and the HIV reversal in the cell culture medium was detected 8 days after infection Recording enzyme activity, compared with the control without IFN-λ, the inhibitory rates of the three IFN-λ inhibitors against HIV were 69.74%, 43.81%, and 76.61%, respectively, and the inhibitory effect of IFN-λ3 was the best. In Fig. 2B, human macrophages were treated with the same dose (100ng/ml) of IFN-λ1, IFN-λ2, and IFN-λ3 for 24 hours, compared with the control group without IFN-λ, the intracellular Spectrum antiviral factor mRNA content increased to varying degrees, and the increase of ISG56, MxA, OAS-1, A3G, and A3F induced by IFN-λ3 was significantly higher than that of IFN-λ1 and IFN-λ2 (p<0.05 ).

The IFN-λ3 used in the present invention was purchased from R&D Systems at a concentration of 100 μg/ml, diluted with phosphate buffered saline (PBS, PH=7.2) to a concentration of 12.5-100 ng/ml for use. The number of human macrophages used is 0.25×106 per well of a standard ²⁴ -well cell culture plate, and the cell culture medium is Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum and 2 mmol/ml glutamine Amide, 100U/ml of penicillin and 100U/ml of streptomycin. When treated with IFN-λ3, 200 microliters of different concentrations of IFN-λ3 were added to the cell culture medium, and the HIV-inhibiting effect shown in Figure 1 and Figure 2 could be observed.

The above is only a preferred embodiment of the present invention, it should be pointed out that, for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications can also be made. It should be regarded as the protection scope of the present invention.

Claims (7)

1. An application of IFN-λ3 in the preparation of a drug for preventing or treating AIDS. 2. The application according to claim 1, wherein the IFN-λ3 acts on the JAK-STAT signaling pathway, regulates signaling molecules, produces broad-spectrum antiviral factors, and inhibits HIV. 3. The application according to claim 2, wherein the broad-spectrum antiviral factors include ISG56, OAS-1, MxA, A3G and A3F. 4. The application according to claim 1, characterized in that the IFN-λ3 inhibits the activity of macrophages in HIV-infected humans. 5. A drug for preventing or treating AIDS prepared from IFN-λ3, characterized in that the purity of the IFN-λ3 is greater than 95%. 6. The medicine according to claim 5, characterized in that the medicine comprises IFN-λ3 and acceptable auxiliary materials. 7. The medicine according to claim 6, characterized in that the concentration of the IFN-λ3 is 12.5-100 ng/ml.