阅读说明：本技术 化合物jq-1用于预防或治疗非洲猪瘟的新用途 (New application of compound JQ-1 in preventing or treating African swine fever ) 是由 牛庆丽 杨吉飞 赵亚茹 刘志杰 关贵全 罗建勋 殷宏 于 2020-07-16 设计创作，主要内容包括：本发明属于非洲猪瘟治疗技术领域,具体涉及一种化合物JQ-1用于预防或治疗非洲猪瘟的新用途。本发明意外发现,化合物JQ-1能够显著抑制ASFV中p30和p72的RNA和蛋白表达水平,阻止病毒入侵宿主细胞,可用于抑制ASFV的早期感染；并且化合物JQ-1能够显著增加ASFV感染后TNF-α、NF-κB、IL-1β、IL-8等基因表达水平,上调TNF-α、NF-κB、IL-1β、IL-8的转录水平,增强免疫反应；因此,化合物JQ-1可用于预防或治疗非洲猪瘟。(The invention belongs to the technical field of African swine fever treatment, and particularly relates to a new application of a compound JQ-1 in preventing or treating African swine fever. The invention unexpectedly discovers that the compound JQ-1 can obviously inhibit the RNA and protein expression levels of p30 and p72 in ASFV, prevent viruses from invading host cells and can be used for inhibiting the early infection of the ASFV; moreover, the compound JQ-1 can remarkably increase the expression levels of genes such as TNF-alpha, NF-kappa B, IL-1 beta, IL-8 and the like after ASFV infection, up-regulate the transcription levels of TNF-alpha, NF-kappa B, IL-1 beta and IL-8 and enhance immune response; therefore, the compound JQ-1 can be used for preventing or treating African swine fever.)

1. The application of a compound JQ-1 in preparing a medicine for treating African swine fever is disclosed, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

2. the application of a compound JQ-1 in preparing a drug for preventing African swine fever is disclosed, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

3. the application of a compound JQ-1 in preparing a medicine for inhibiting transcription and expression of an African swine fever virus gene is disclosed, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

4. the application of a compound JQ-1 in preparing a medicament for inhibiting the expression of African swine fever virus p30 RNA, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

5. the application of a compound JQ-1 in preparing a medicine for inhibiting the expression of the p30 protein of the African swine fever virus is disclosed, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

6. the application of a compound JQ-1 in preparing a medicament for inhibiting the expression of African swine fever virus p72 RNA, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

7. the application of a compound JQ-1 in preparing a medicine for inhibiting the expression of the p72 protein of the African swine fever virus is disclosed, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

8. the application of a compound JQ-1 in preparing a medicament for promoting the expression of inflammatory factors, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

9. the use of claim 8, wherein said inflammatory factor comprises TNF- α, NF- κ B, IL-1 β, IL-8.

10. The use of any one of claims 1 to 9, wherein the compound JQ-1 is formulated into any one of a tablet, a spray, a granule, a capsule, an oral liquid, an injection, and a suspension by adding a pharmaceutically acceptable carrier and/or adjuvant.

Technical Field

The invention belongs to the technical field of African swine fever treatment, and particularly relates to a new application of a compound JQ-1 in preventing or treating African swine fever.

Background

African Swine Fever (ASF) is an acute virulent infectious disease characterized by Fever of pigs and organ bleeding of the whole body of pigs caused by African Swine Fever Virus (ASFV), and the death rate of domestic pigs is as high as 100%. The disease first outbreaks in kenya 1921 and then is widely prevalent in domestic and wild pigs throughout africa. The 20 th century was introduced into europe in the 50 s, and the disease was cured for 40 years throughout europe. However, the disease was again introduced into grurgia from eastern africa in 2007, and then widely disseminated in eastern europe and introduced into elocusk, the far east russia, 2017. At the beginning of 8 months in 2019, a Hurongrong researcher reports the epidemic situation of the African swine in the first instance of China, and the disease spreads to 30 provinces and municipalities in China within a short time of one year, so that the disease continues to threaten the pig industry, wherein compared with 8 months in 2018, the yield of the domestic pig in 9 months in 2019 is reduced by 40%, the price of pork is doubled since 8 months in 2019, the yield is reduced by more than 40% in China, and the loss is serious. As no effective vaccine or specific therapeutic medicine exists so far, once the epidemic situation of the African swine fever occurs, the epidemic situation can be controlled only by a killing means, but the mode not only causes economic loss, but also cannot meet the requirement of large-scale pig raising in China. Therefore, how to effectively control the ASF epidemic situation is one of the great challenges facing the pig industry in the world at present, and is also a major strategic subject to be urgently solved by ASF prevention and control in China.

The p30 and p72 are key structural proteins in ASFV, and can neutralize virus after the virus attacks susceptible cells, and inhibit the attachment, replication and internalization processes of the virus. Wherein p30 is the main structural protein constituting the virus particle, and is also an important surface antigen, and is closely related to host cell tropism, pathogenicity and immunogenicity; p30 is expressed early in ASF infection, usually produced 2-4h post infection, and is expressed continuously throughout the infection, involved in virus internalization, and is involved in virus invasion into host cells. And p72 is a capsid protein, is the main structural protein of the African swine fever virus, can protect the virus nucleic acid from being damaged by nuclease or other physicochemical factors in the environment, is involved in the infection process of the virus and has good immunogenicity, the protein is generated in the late stage of virus infection, p72 is an important antigen protein of ASFV, is the main component of virus icosahedron, is important for the formation of virus capsid, and is involved in virus binding cells.

The compound JQ-1 is a small molecule compound that binds directly to the Kac binding site of the BET bromopain domain. The existing research shows that: JQ1(500nM) acted on NMC797 cells, effectively reducing the expression of BRD4 target gene. Acts on NMC 11060 cells to inhibit cell activity. JQ1(500nM) acted on LP-1 cells, inhibiting MYC, BRD4 and CDK9 expression. JQ1 (1. mu.M) treated latently infected Jurkat T cells to activate HIV transcription. In vivo studies showed that JQ1(50mg/kg) treated mice bearing NMC797 transplanted tumors inhibited tumor growth. JQ1(50mg/kg)) wiped off NUT nuclear spots in mice bearing NMC797 transplanted tumors, consistent with competitive binding to chromatin. (+) -JQ1(50mg/kg) treated a mouse model carrying NMC transplanted tumors, promoting differentiation, tumor regression, and prolonging life.

The invention unexpectedly discovers that the compound JQ-1 can obviously inhibit the RNA and protein expression levels of p30 and p72 in ASFV, prevent viruses from invading host cells and can be used for inhibiting the early infection of the ASFV; moreover, the compound JQ-1 can remarkably increase the expression levels of genes such as TNF-alpha, NF-kappa B, IL-1 beta, IL-8 and the like after ASFV infection, up-regulate the transcription levels of TNF-alpha, NF-kappa B, IL-1 beta and IL-8 and enhance immune response; therefore, the compound JQ-1 can be used for preventing or treating African swine fever.

Disclosure of Invention

In view of the above technical problems, the present invention aims to provide an application of a compound JQ-1 in preparing a drug for treating african swine fever, wherein the structural formula of the compound JQ-1 is shown as the following formula (i):

the invention also aims to provide an application of a compound JQ-1 in preparing a medicine for preventing African swine fever, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

the invention also aims to provide an application of a compound JQ-1 in preparing a medicine for inhibiting transcription and expression of African swine fever virus genes, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

the invention also aims to provide an application of a compound JQ-1 in preparing a medicament for inhibiting the expression of African swine fever virus p30 RNA, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

the invention also aims to provide an application of a compound JQ-1 in preparing a medicine for inhibiting the expression of the African swine fever virus p30 protein, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

the invention also aims to provide an application of a compound JQ-1 in preparing a medicament for inhibiting the expression of African swine fever virus p72 RNA, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

the invention also aims to provide an application of a compound JQ-1 in preparing a medicine for inhibiting the expression of the African swine fever virus p72 protein, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

the invention also aims to provide an application of a compound JQ-1 in preparing a medicament for promoting the expression of inflammatory factors, wherein the structural formula of the compound JQ-1 is shown as the following formula (I):

preferably, the inflammatory factors include TNF-alpha, NF-kappa B, IL-1 beta, IL-8.

Preferably, the compound JQ-1 is added with a pharmaceutically acceptable carrier and/or an auxiliary material to be prepared into any one dosage form of tablets, sprays, granules, capsules, oral liquid, injections and suspensions.

The invention has the beneficial effects that: the invention unexpectedly discovers that the compound JQ-1 can obviously inhibit the RNA and protein expression levels of p30 and p72 in ASFV, prevent viruses from invading host cells and can be used for inhibiting the early infection of the ASFV; moreover, the compound JQ-1 can remarkably increase the expression levels of genes such as TNF-alpha, NF-kappa B, IL-1 beta, IL-8 and the like after ASFV infection, up-regulate the transcription levels of TNF-alpha, NF-kappa B, IL-1 beta and IL-8 and enhance immune response; therefore, the compound JQ-1 can be used for preventing or treating African swine fever.

Drawings

FIG. 1 Compound JQ-1 inhibits African swine fever virus infection and replication;

FIG. 2 is a graph showing the results of the RNA expression levels of p30 and p 72;

FIG. 3 is a graph showing the results of the compound JQ-1 inhibiting the protein expression levels of African swine fever viruses p30 and p 72;

FIG. 4 is a graph showing the results of up-regulation of host inflammation-related factor expression by JQ-1;

FIG. 5 is a graph showing the cytotoxicity results of JQ-1 compound.

Detailed Description

In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the specific embodiments. The scope of the invention is not limited to the examples described below.

The experiments described in the following examples obtain biosafety permits and african swine fever laboratory activity permits:

according to the related requirements of biosafety of a Lanzhou veterinary research institute of the Chinese agricultural academy of sciences, a biological safety 3-level laboratory (BSL-3) and related biological safety of African swine fever, the Lanzhou veterinary research institute biological safety committee, the laboratory animal ethics committee, the Chinese agricultural academy of sciences biological safety committee, the Lanzhou veterinary research institute experimental animal ethics committee and the Lanzhou veterinary research institute biological safety committee report step by step, the permission of developing highly pathogenic ASFV pathogens and animal research is obtained by the agricultural department, and the permission is recorded by the agricultural rural department and meets the requirements of national biological safety level.

Experimental cells, viral sources as described in the examples below:

primary Porcine Alveolar Macrophages (PAM) and primary bone marrow macrophages (BMDM) were taken from healthy SPF Bama minipigs aged 2-4 months, aseptically collected, lysed with red blood cell lysate (purchased from Biosharp), red blood cells were removed, centrifuged at low speed, the supernatant was discarded, and the cell pellet was resuspended in RPMI 1640 complete medium (purchased from Gibco) containing 10% FBS (purchased from PAN), placed at 37 ℃ and 5% CO₂Culturing in an incubator.

The ASFV genotype II strain (ASFV CN/SC/2019) is an African swine fever regional laboratory isolate of Lanzhou veterinary research institute of Chinese academy of agricultural sciences, and the virus titer is 5 × 10⁷TCID₅₀and/mL, which is the 4 th generation virus after PAM cell propagation, is subpackaged and stored in a foot and mouth disease reference laboratory (ABSL-3) of China and is reserved at the temperature of 80 ℃ below zero for later use.

JQ-1, purchased to sigma aldrich trade company, brand: sigma, cat No.: SML1524-5 MG.

Other reagents in the experiment are common commercial reagents unless otherwise specified; the procedures in the experiments are those known in the art unless otherwise specified.