阅读说明：本技术 甘草苷在制备治疗或预防肠道病毒71型感染药物中的应用 (Application of liquiritin in preparation of medicine for treating or preventing enterovirus71 infection ) 是由 李晋 徐尚福 杜文琪 于 2021-09-09 设计创作，主要内容包括：本申请公开了中药制药技术领域中甘草苷在制备治疗或预防肠道病毒71型感染药物中的应用以及制备抑制肠道病毒71型复制药物中的应用,研究发现,本申请采用甘草苷,灌胃给药后,可通过降低脑组织中EV71VP1表达量,降低炎症因子IL-6,抑制病毒复制,进而改善EV71引起手足口病的神经症状。(The application discloses application of liquiritin in preparing medicines for treating or preventing enterovirus71 type infection and application of liquiritin in preparing medicines for inhibiting enterovirus71 type replication in the technical field of traditional Chinese medicine pharmacy, and researches show that liquiritin is adopted in the application, after the liquiritin is administrated by intragastric administration, EV71VP1 expression quantity in brain tissues can be reduced, inflammatory factor IL-6 is reduced, virus replication is inhibited, and then the neurological symptoms of hand-foot-and-mouth disease caused by EV71 are improved.)

1. Application of liquiritin or pharmaceutically acceptable salt thereof as an active ingredient in preparation of medicines for treating or preventing enterovirus71 infection.

2. Application of liquiritin or pharmaceutically acceptable salt thereof as an active ingredient in preparing a medicament for inhibiting replication of enterovirus 71.

3. Use according to claim 1, characterized in that: the diseases caused by enterovirus71 infection comprise one or more of hand-foot-and-mouth disease, viral angina, aseptic meningitis, brainstem encephalitis, neurogenic pulmonary edema and central nervous system infection.

4. The use of claim 3, wherein the medicament is a tablet, a capsule, an oral liquid, an oral granule, an oral powder or an injection.

Technical Field

The invention relates to the technical field of traditional Chinese medicine pharmacy, relates to application of liquiritin in preparation of antiviral drugs, and particularly relates to application of liquiritin in preparation of drugs for treating or preventing enterovirus71 infection.

Background

Enterovirus type 71 (enterovirus71, EV71) infection has become a major threat to global public health, especially in infants and young children. Epidemiological studies have shown that EV71 infection is the leading cause of severe and even fatal hand-foot-and-mouth disease. EV71 infection is often accompanied by serious neurological complications such as aseptic meningitis, acute flaccid paralysis, encephalitis, etc., which are considered the major disease process in fatal cases, and neurogenic pulmonary edema. For central nervous system diseases caused by EV71 infection, no effective antiviral therapeutic drugs and vaccines applicable to clinical application exist. Therefore, the active development of drugs having anti-EV 71 effects and having protective effects on the nervous system is urgently needed.

The existing research of resisting EV71 by traditional Chinese medicines shows that preparations such as qingkailing, Xiyanping, and radix scutellariae oral liquid, and single traditional Chinese medicines such as coltsfoot and houttuynia cordata have the effect of resisting EV71, but the specific antiviral components and mechanism discussion are still insufficient. However, the pathogenesis of central nervous system damage caused by EV71 infection and the unclear mechanism of antiviral or neuroprotective action of traditional Chinese medicines or natural products are bottlenecks that restrict the safe and effective treatment. Therefore, the antiviral active ingredients of the traditional Chinese medicine are determined, the action mechanism of the antiviral active ingredients is proved, the antiviral active ingredients are one of important ways for the prevention and treatment research of the central nervous system diseases related to EV71, and the basic research basis is provided for the traditional Chinese medicine treatment of the central nervous system diseases caused by EV71 infection.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides the application of liquiritin in resisting enterovirus 71.

One of the purposes of the invention is to provide the application of liquiritin or pharmaceutically acceptable salts thereof as an active ingredient in preparing a medicament for treating or preventing enterovirus71 infection.

The invention also aims to provide application of liquiritin or pharmaceutically acceptable salt thereof as an active ingredient in preparing a medicament for inhibiting enterovirus71 replication.

Specifically, the diseases caused by the enterovirus71 infection comprise one or more of hand-foot-and-mouth disease, viral angina, aseptic meningitis, brainstem encephalitis, neurogenic pulmonary edema and central nervous system infection.

Specifically, the medicament can be prepared into any dosage form, including but not limited to the following: tablets (e.g., coated tablets), capsules, oral liquids, oral granules, oral powders, and injections (e.g., lyophilized powders and emulsions for injection).

The liquiritin is sold on the market, and researches show that after the liquiritin is used for gastric administration, the liquiritin can reduce the expression level of EV71VP1 in brain tissues, reduce inflammatory factors IL-6, inhibit virus replication, further improve hand-foot-and-mouth disease or other neurological symptoms caused by EV71 and achieve the purpose of preventing and treating diseases caused by EV71 infection.

Drawings

FIG. 1 is a graph of body weight change, survival and clinical symptom scores for each group in a suckling mouse infection experiment;

FIG. 2 is a graph showing the results of HE staining of brain tissues of each group;

FIG. 3 is a diagram showing the expression level of EV71VP1 in each group of brain tissues;

FIG. 4 is a diagram showing the expression level of IL-6, an inflammatory factor in brain tissue in each group.

Detailed Description

The following is further detailed by way of specific embodiments:

1. establishing an EV71 infected suckling mouse model, and observing the treatment effect of liquiritin on EV71 infected suckling mice

1.1 Molding method

30 ICR suckling mice with 3 days old weigh 3 +/-0.5 g and are divided into 3 groups, and 10 mice in each group are respectively a first blank Control group (Control group); model group (EV71 group); ③ liquiritin treatment group (LQ group). The model group and the liquiritin treatment group were administered with 50 μ L intracranial injection of EV71 virus stock solution to prepare an EV71 infection model, and the blank control group was administered with an equivalent amount of saline intracranial injection. On the 2 nd day of molding, the liquiritin treatment group adopts liquiritin 0.06mg/g for intragastric administration, 1 time per day, for 5 days. The blank control group and the model group were subjected to the same dose of saline for intragastric administration.

1.2 weight change, survival and clinical symptom scores

Body weight of the suckling mice was measured, and changes in body weight before and after administration were observed. Clinical symptom scores were made for each group of suckling mice, and the clinical scoring criteria are shown in the table below.

Table 1 shows the clinical scoring criteria

1.3 brain histopathological examination

After the administration on day 5, the suckling mice were fasted and anesthetized, and the brain tissue was taken, subjected to HE staining, and observed for pathological changes in the brain tissue.

1.4 detection of brain tissue EV71VP1 expression level

After the administration on day 5, the suckling mice were fasted and anesthetized, and brain tissue was taken, RNA was extracted, and the expression level of EV71VP1 gene in brain tissue was examined.

1.5 detection of expression level of inflammatory factor IL-6 in brain tissue

After the administration on day 5, the suckling mice were fasted and anesthetized to decapitate, brain tissue was taken, RNA was extracted, and the expression of IL-6 in the brain tissue was examined.

2 results of the experiment

2.1 weight change, survival and clinical symptom scores

After 3 days old suckling mice were inoculated with virus, weight change, survival and clinical symptom scores were recorded daily. In particular, as shown in fig. 1, a is a gross comparison between the model group suckling mice and LQ-treated suckling mice; b is the change curve of the body weight of the suckling mice in each experimental group; c is survival rate curve of the suckling mice in each experimental group; d is the clinical score of each experimental group. Compared with a blank control group, the weight of the suckling mice in the model group is obviously reduced, the survival rate is slightly reduced, and the clinical score is obviously increased. After LQ administration treatment, the weight of the suckling mice is obviously increased, the survival rate is improved, the clinical score is obviously reduced,

2.2 pathological examination of brain tissue

Under the microscope, as shown in fig. 2, a is a blank control group, B is a model group, and C is an LQ treatment group. The blank control group had a large number of nerve cells in the cerebral cortex and had round and moist cells. The nerve cell morphology of the model group is shriveled and a large amount of inflammatory cell infiltration occurs, and after the LQ administration treatment, the nerve cell morphology is improved compared with the model group, and the inflammatory cell infiltration is reduced.

2.3 detection result of brain tissue EV71VP1 expression level

EV71 belongs to picornaviridae enterovirus (enterovirus), which contains a single-stranded positive-stranded RNA genome of about 7kb in length. During viral infection, four capsid proteins (VP1, VP2, VP3 and VP4) are translated, of which VP1 plays a dominant role in the acquisition and transmission of the virus. As shown in fig. 3, vs blank, # P < 0.05; vs model group, P < 0.05. In the blank control group, EV71VP1 was not expressed, whereas model group EV71 VP1mRNA was highly expressed. Compared with the model group, the EV71 VP1mRNA of the liquiritin treatment group is obviously reduced.

2.4 detection results of IL-6 expression level of inflammatory factor in brain tissue

IL-6 is a pleiotropic cytokine produced by immune and non-immune cells that stimulates the immune response of the body. As shown in fig. 4, vs blank, # # P < 0.01; vs model group,. P < 0.05; compared with a normal control group, the IL-6mRNA expression of the model group is obviously reduced, and the IL-6mRNA expression is obviously increased after LQ administration. The LQ is shown to promote the immunocompetence of body cells and enhance the resistance of the body cells to EV 71.

The foregoing is merely an example of the present invention and common general knowledge of known specific structures and features of the embodiments is not described herein in any greater detail. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.