阅读说明：本技术 氯离子通道抑制剂兰雪醌在抗腹泻药物中的应用 (Application of chloride channel inhibitor lanjiquinone in anti-diarrhea drugs ) 是由 于波 杨红 朱小娟 于 2019-11-05 设计创作，主要内容包括：本发明涉及氯离子通道抑制剂兰雪醌在抗腹泻药物中的应用。肠道Cl<Sup>-</Sup>通道作为肠道液体分泌的关键通道,已经成为了一类很有潜力的腹泻治疗靶点。本发明涉及兰雪醌对肠道Cl<Sup>-</Sup>通道抑制活性的发现,特别是口服兰雪醌对细菌和病毒诱导腹泻的治疗作用。研究表明,兰雪醌可逆地抑制了TMEM16A通道活性,对CaCC和CFTR Cl<Sup>-</Sup>通道也具有抑制作用。灌胃兰雪醌可降低STa和CT诱导的肠液分泌,并显著改善霍乱毒素和轮状病毒诱导的腹泻症状。此外,肠动力研究显示,兰雪醌能够显著减缓肠蠕动,抑制肠平滑肌收缩,但对收缩频率无明显影响。本发明所述的兰雪醌可应用于治疗霍乱、旅行者腹泻和轮状病毒腹泻的药物中,开拓了兰雪醌的新药理学活性,为抗腹泻药物的研发提供了新的选择。(The invention relates to application of a chloride ion channel inhibitor lanjiquinone in an anti-diarrhea drug. Intestinal canal Cl ‑ The channel is used as a key channel for intestinal fluid secretion and becomes a potential diarrhea treatment target. The invention relates to Lanceoquinone p-intestinal Cl ‑ Discovery of channel inhibitory activity, particularly therapeutic effects of oral lanugo quinone on bacterial and viral induced diarrhea. Studies have shown that lanugenon reversibly inhibits TMEM16A channel activity against CaCC and CFTR Cl ‑ The channels also have an inhibitory effect. The gavage lanuginose can reduce intestinal juice secretion induced by STa and CT, and obviously improve diarrhea symptoms induced by cholera toxin and rotavirus. In addition, intestinal motility studies have shown thatThe snow quinone can obviously slow down intestinal peristalsis and inhibit contraction of intestinal smooth muscle, but has no obvious influence on contraction frequency. The lanugenone can be applied to medicines for treating cholera, traveler's diarrhea and rotavirus diarrhea, develops new pharmacological activity of the lanugenone, and provides a new choice for research and development of anti-diarrhea medicines.)

1. An application of a chloride channel inhibitor lanugo quinone in preparing anti-diarrhea drugs is characterized in that the compound lanugo quinone shown in formula (I) is applied in preparing anti-bacterial diarrhea or viral diarrhea drugs;

2. the application of the chloride ion channel inhibitor lanjiquinone in the preparation of medicines for treating intestinal motility disorder is characterized in that the compound lanjiquinone shown in the formula (I) is applied in the preparation of medicines for treating intestinal motility disorder;

3. the use according to claim 1, wherein the diarrhea comprises cholera toxin-induced diarrhea or rotavirus diarrhea.

4. Use according to claim 1 or 2, characterized in that the compound and the pharmaceutically acceptable salts thereof are formulated as an oral preparation for the treatment of diarrhea and intestinal motility disorder-related diseases.

Technical Field

The invention relates to the field of medicines, in particular to an inhibitory effect of lankeequinone on chloride ion channels and application thereof in treatment of bacterial and viral diarrhea.

Background

Diarrhea is a serious health-threatening disease, with high morbidity and mortality worldwide, particularly in children and the elderly. According to the incomplete statistics of 2015, about 577,000 children under 5 years of age and 502,000 elderly over 70 years of age die worldwide from secretory diarrhea. In developing countries, the enteropathogenic bacteria causing diarrhea are mainly vibrio cholerae and pathogenic escherichia coli, and the pathogenic viruses are mainly rotavirus and enteroinvasive virus.

The intestinal tract is an important digestive organ of the human body, and the main physiological functions relate to the physiological processes of substance absorption, intestinal fluid secretion, intestinal movement and the like. The flow of fluid between the intestine and the blood is mainly influenced by Na⁺、Cl^-、HCO₃ ^-And K⁺And driving of solutes such as glucose. Intestinal fluid secretion is mainly Cl passing through the basal and apical membranes^-Channel and transporter pairs Cl^-Is achieved by secretion. Cl involved in intestinal fluid secretion^-The channels are mainly cystic fibrosis transmembrane conductance regulator (CFTR) and calcium activated chloride channel (CaCC). Cholera toxin secreted by vibrio cholerae and enterotoxigenic escherichia coli and heat-resistant escherichia coli endotoxin can increase intracellular cAMP, cGMP and Ca²⁺And both of these molecules can activate CFTR activity. The bacteria also can increase the levels of various receptor agonists, neurotransmitters and neuropeptide receptors in body fluids, such as 5-hydroxytryptamine (5-HT), Vasoactive Intestinal Peptide (VIP) and galanin type I receptors, thereby activating intestinal cells to Cl^-Secretion of (5). Rotavirus infection can cause fluid secretion and changes in the epithelial structure of the intestinal tract, causing age-related secretory diarrhea. About 50 million infants die of rotavirus diarrhea every year in the world, and in China, infants hospitalized due to rotavirus infection account for about 40 percent of all infants hospitalized due to acute gastrointestinal diseases. Current studies indicate that the non-structural protein (NSP4) produced by rotavirus causes intracellular Ca by binding to the membrane receptor (integrin. alpha.1beta.2), or by binding to the receptor via glycerol peptide, or by stimulating the enteric nerve²⁺Increased concentration, Cl production by CaCC on the cell surface^-It is secreted to cause diarrhea. In addition to the liquid secretion mechanism, Cl^-The channels are also involved in the regulation of intestinal motility. TMEM16A, a known CaCC, has high expression in Interstitial Cajal Cells (ICC) in the intestinal tract. The ICC has the main function as a pacing point and is involved in the generation and conduction of gastrointestinal electrical slow waves, and constitutes a link between sensory nerves and smooth muscle cells. The normal contraction function of the intestinal muscles is mainly dependent on the coordination activity of the Enteric Nervous System (ENS) and ICC, and various digestive tract dyskinetic diseases are closely related to ICC abnormalities, such as: megacolon, constipation, abdominal pain and constipation which are frequently occurred in patients with long-term insulin-dependent diabetes mellitus. The study shows that TMEM16A protein has specific expression in ICC of all parts of gastrointestinal tracts of birds, non-human primates and humans, and the CaCC blocking agents niflumic acid and DIDS can weaken contraction frequency of stomach and small intestine in a dose-dependent mode and block slow wave generation. The selective inhibitor of TMEM16A can obviously weaken the contraction of smooth muscle of intestinal segment of mouse, while the activator of TMEM16A can enhance the contraction and restore the contraction force of small intestine after atropine inhibition. The lack of function of TMEM16A prevented the mouse gastrointestinal tract from forming slow waves, but did not affect the formation of the ICC network. The above results demonstrate the fundamental role of TMEM16A in the generation of slow waves in the gastrointestinal tract and in the regulation of smooth muscle contractility.

In view of Cl^-Channel activation may be a common mechanism for the development of multiple diarrheas, and therefore screening against different Cl's from combinatorial chemical small molecule libraries^-The channel inhibitors become hot spots of research on treatment and prevention of diarrhea, and a plurality of small molecule inhibitors belonging to different structural families are discovered at present, and part of inhibitors are applied to clinical disease treatment.

Disclosure of Invention

In order to make up for the blank of the prior art, the invention provides the application of a chloride channel inhibitor lanugo quinone (plumbagin) in the preparation of anti-diarrhea drugs and drugs for intestinal motility disorder diseases.

The invention has the following inventive concept: the natural small molecules have the characteristics of various structures, high bioactivity, relatively stable structure, high bioavailability, small toxic and side effects and the like, the inventor discovers the lanugenon from a natural small molecule library, and the lanugenon inhibits Cl at the apical membrane side of the FRT cell transfected with TMEM16A^-Current, which has TMEM16A inhibitory activity. The regulation activity of the lanugenone on CFTR and intestinal epithelial CaCC is firstly researched in vitro by using various means, the intestinal fluid secretion and intestinal power regulation of the lanugenone are systematically analyzed by using isolated mouse intestinal tissues, and finally the diarrhea resistance effect of the lanugenone is proved on various mouse diarrhea models, so that the lanugenone has higher application value in the aspect of clinical diarrhea resistance treatment.

The compound lancedrone has the following structure:

the experiment of the invention proves that the natural Cl^-Lancetone, a channel inhibitor, has inhibitory effects on CaCC and CFTR activity on HT-29 cells in colon cancer.

Lancequinone reduces intestinal fluid secretion induced by E.coli heat stable enterotoxin (STa) and Cholera Toxin (CT).

The diarrhea inhibiting rate of the lanugo quinone is more than 40% in a cholera toxin newborn mouse diarrhea model, and is 50% in a rotavirus-induced newborn mouse diarrhea model.

The lancedrone obviously inhibits the movement of the intestinal tract and increases the absorption time of intestinal tract liquid; inhibit intestinal smooth muscle contraction force, but has no obvious effect on contraction frequency.

Therefore, the lanugenone and the pharmaceutically acceptable salts thereof are prepared into oral preparations and used for preparing the medicines for resisting diarrhea and intestinal motility disorder-related diseases.

Such diseases or conditions include: treating intestinal motility disorder, diarrhea caused by cholera toxin or rotavirus diarrhea.

The invention discovers a novel pharmacological activity of the lanugenone, namely, by inhibiting Cl^-The channel activity can inhibit intestinal fluid secretion and intestinal peristalsis enhancement induced by intestinal pathogens. Lanceoquinone can be used as a lead compound for treating cholera toxin-induced diarrhea, traveler's diarrhea, rotavirus diarrhea and other types of secretory diarrhea.

Drawings

FIG. 1 short circuit current measurement of inhibition of TMEM16A by lancedrone;

FIG. 2 short circuit current measurement of the inhibitory effect of lankeequinone on the intestinal epithelium CaCC and CFTR;

FIG. 3 Lancequinone inhibits the secretion of intestinal fluids induced by STa and CT in isolated intestinal tracts;

FIG. 4 Lancequinone inhibits cholera toxin and rotavirus infection induced neonatal mouse diarrhea;

FIG. 5 shows the inhibitory effect of lancedrone on intestinal motility.

Detailed Description

In order to better explain the technical solution of the present invention, the following will describe the embodiments of the present invention in further detail with reference to examples. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The langjiquinone standard used in the present invention was purchased from Dalian Meilun Biotechnology Ltd. The specific information is as follows: the purity HPLC of the lanthaquinone is more than or equal to 98 percent; the molecular formula is as follows: C11H8O 3; molecular weight: 188.18.