Application of Bunazosin Hydrochloride in Preventing and Treating Intestinal Ischemia-Reperfusion Injury

Abstract

The present invention belongs to the field of biotechnology, and specifically relates to the application of bunazosin hydrochloride in preventing and treating intestinal ischemia-reperfusion injury. The bunazosin hydrochloride is a bunazosin hydrochloride solution; the bunazosin hydrochloride prevents and treats intestinal ischemia-reperfusion injury by reducing intestinal ischemia-reperfusion injury, inhibiting damage to extraintestinal organs, increasing the survival time of model animals, inhibiting oxidative stress and apoptosis induced by ischemia-reperfusion, and promoting epithelial cell proliferation. The inventor has experimentally confirmed that bunazosin can effectively reduce intestinal ischemia-reperfusion injury, inhibit damage to extraintestinal organs, significantly increase the survival time of model animals, inhibit oxidative stress and apoptosis induced by ischemia-reperfusion, and promote the proliferation of epithelial cells, which provides a basis for the screening of new drugs for the treatment of intestinal ischemia-reperfusion injury.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to application of bunazosin hydrochloride in preventing and treating intestinal ischemia-reperfusion injury. Background Art

[0002] Intestinal ischemia / reperfusion (I / R) is a common condition in a variety of clinical situations such as trauma, shock, infection, intestinal obstruction and extracorporeal circulation. Although acute intestinal ischemia is reversible with early treatment, blood reperfusion will further aggravate intestinal tissue damage. In addition, intestinal tissue damage will increase the permeability of the intestinal barrier, and a large number of bacteria, endotoxins, inflammatory factors, etc. will be released into the blood, leading to damage or even failure of extraintestinal organs.

[0003] During ischemia, intestinal tissue begins to be damaged due to lack of oxygen and nutrients. The mitochondrial dysfunction of the cells leads to reduced cellular energy, which affects cellular homeostasis. In addition, ischemia can also lead to increased intracellular calcium concentration, acidosis, increased free radical production, and the release of inflammatory mediators. Although ischemia itself can cause cell damage, the real damage often occurs when blood flow is restored, that is, during the reperfusion phase. When blood flow is restored, a large amount of reactive oxygen and nitrogen species are produced, inducing oxidative stress, which damages cell structure and function. In addition, activated neutrophils will accumulate in ischemic tissues and release inflammatory factors and proteases, further aggravating tissue damage. This generation of reactive oxygen and nitrogen species, together with the activation of neutrophils, leads to increased vascular permeability and the formation of edema. Reperfusion injury also affects the intestinal mucosal barrier, which may lead to the infiltration of bacteria and toxins into the circulation, causing systemic infection or multiple organ failure. Therefore, it is crucial to study the mechanisms of intestinal I / R injury and find possible treatment strategies.

[0004] In order to study the mechanism of intestinal I / R injury, scientists have developed a variety of animal models. The most common method is to induce ischemia by clamping the superior mesenteric artery or its branches, and then release the arterial clamp to restore blood flow. In this way, the intestinal ischemia and reperfusion process in the human body can be simulated. In terms of treatment, researchers have explored a variety of potential strategies, such as free radical scavengers, anti-inflammatory drugs, calcium channel antagonists, and mitochondrial protectors. Among them, some have shown protective effects against intestinal I / R injury in experimental animals. Currently, the early diagnosis of intestinal I / R injury remains challenging due to the ambiguity of clinical manifestations and the lack of early specific biomarkers. Although intestinal fatty acid binding protein (I-FABP) and smooth muscle protein (SM22) are considered as markers for detecting intestinal injury, they are still limited in specificity and sensitivity. Therefore, it is crucial to find more specific and sensitive markers of intestinal injury.

[0005] In general, intestinal ischemia-reperfusion injury is a complex disease involving the interaction of multiple cells and molecules. Understanding its mechanism and finding effective treatment strategies are the focus of current research. Summary of the invention

[0006] The purpose of the present invention is to provide an application of bunazosin hydrochloride in preventing and treating intestinal ischemia-reperfusion injury, effectively improving intestinal ischemia-reperfusion injury indicators and improving prognosis.

[0007] The objective of the present invention is achieved through the following technical solutions:

[0008] The invention provides an application of bunazosin hydrochloride in preventing and treating intestinal ischemia-reperfusion injury.

[0009] Furthermore, the bunazosin hydrochloride is a bunazosin hydrochloride solution.

[0010] Furthermore, the bunazosin hydrochloride prevents and treats intestinal ischemia-reperfusion injury by alleviating intestinal ischemia-reperfusion injury, inhibiting damage to extraintestinal organs, increasing the survival time of model animals, inhibiting ischemia-reperfusion-induced oxidative stress and apoptosis, and promoting epithelial cell proliferation.

[0011] The present invention also provides an application of bunazosin hydrochloride in preparing a medicine for preventing and treating intestinal ischemia-reperfusion injury.

[0012] Furthermore, the bunazosin hydrochloride can be added with pharmaceutically acceptable excipients to prepare any dosage form including injection, capsule, tablet, granule, suspension, emulsion, spray, powder, liposome, oral solution, and pill.

[0013] Furthermore, the administration method of the drug includes one or more of intramuscular injection, subcutaneous injection, intravenous injection, oral administration, sublingual administration, intralesional or intracerebral or implanted delivery, and spray administration.

[0014] The beneficial effects of the present invention are as follows: the inventors have experimentally confirmed that bunazosin can effectively reduce intestinal ischemia-reperfusion injury, while inhibiting damage to extraintestinal organs, significantly increasing the survival time of model animals, inhibiting ischemia-reperfusion-induced oxidative stress, apoptosis and the ability to promote epithelial cell proliferation, thus providing a basis for the screening of new drugs for the treatment of intestinal ischemia-reperfusion injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The figures are the experimental results of the in vivo experiment in Example 1 to verify the effect of bunazosin hydrochloride on preventing and treating intestinal ischemia-reperfusion injury, wherein Figure A is the HE pathological score of small intestinal injury in different model mice; Figure B is the survival rate of different model mice; and Figure C is the endotoxin level in the plasma of different model mice. DETAILED DESCRIPTION

[0016] The specific product information of bunazosin hydrochloride used in the present invention is as follows:

[0017] Product Name: Bunazosin Hydrochloride

[0018] Molecular formula: C 19 H 28 ClN5O3

[0019] Molecular weight: 409.91

[0020] This drug is a sustained-release dosage form. Preparation features: The tablets are compressed from multiple sustained-release particles of bunazosin hydrochloride and excipients. Each particle is coated with a hydrophobic film. After oral administration, the tablets quickly decompose in the stomach, allowing the sustained-release particles to diffuse and slowly and continuously release the active ingredients into the gastrointestinal tract. Oral absorption is good and no effect of diet was found. Relative bioavailability is 81.1%. The peak blood drug concentration is 10 ng / ml, and the effective blood drug concentration (4-5 ng / ml) can be maintained for 12-14 hours. T1 / 2 is about 12 hours, and it is mainly metabolized in the liver.

[0021] The preparation method of bunazosin hydrochloride in the present invention is as follows:

[0022] Method 1: 4-amino-2-chloro-6,7-dimethoxyquinazoline and 1-formyl homopiperazine are refluxed in butanol; the resulting compound (I) is refluxed in 9% hydrochloric acid to obtain compound (II) with a yield of 80.4%. It is then reacted with butyryl chloride in acetone to obtain bunazosin.

[0023] Method 2: 2-amino-4,5-dimethoxybenzonitrile, compound (III) and sodium methoxide are refluxed in dimethyl sulfoxide to obtain bunazosin.

[0024] Among them, bunazosin can be added with pharmaceutically acceptable excipients to prepare one of the following dosage forms: injection, capsule, tablet, granule, suspension, emulsion, spray, powder, liposome, oral solution, and pill. The preferred dosage form is injection.

[0025] The administration method of bunazosin is one or more of intramuscular injection, subcutaneous injection, intravenous injection, oral administration, sublingual administration, intralesional or intracerebral or implanted delivery, and spray administration, preferably oral, intramuscular, subcutaneous or intravenous injection.

[0026] Example 1 In vivo experiments verify that bunazosin hydrochloride solution reduces intestinal ischemia-reperfusion injury

[0027] At the animal level, the intestinal ischemia-reperfusion injury (I / R) model was established using C57BL / 6J mice by intragastric gavage pretreatment with bunazosin hydrochloride solution, and the survival rate of the mice was detected; the HE pathological damage of intestinal tissue, intestinal barrier permeability, and serum endotoxin (LPS) levels confirmed that bunazosin hydrochloride solution can alleviate intestinal ischemia-reperfusion injury (I / R).

[0028] 1. Establishment of intestinal ischemia-reperfusion injury model:

[0029] To establish the intestinal I / R model, C57BL / 6J mice were fasted for 12 hours before surgery and allowed to drink water freely. General anesthesia was performed using 4% isoflurane, and the surgical anesthesia concentration was maintained at 1.5%. The mice were fixed in a supine position on the operating table, and the abdomen was shaved and disinfected with 75% ethanol. A 1-3 cm laparotomy was performed through the midline of the abdomen, and the abdominal cavity was entered layer by layer. Subsequently, the superior mesenteric artery was clamped to block blood flow for 1 hour. After that, the blood supply was restored and there was no bleeding in the abdominal cavity, and the abdominal cavity was sutured layer by layer. Each operation was accompanied by a subcutaneous injection of 0.5 ml of 37°C saline. After 2 hours of reperfusion, the inferior vena cava blood and each intestinal segment tissue were removed. The operation of the sham group (Sham) was the same as that of the I / R group, but the artery was not clamped.

[0030] 2. Experimental groups:

[0031] Twenty-four 6-8 week old C57BL / 6J mice were used in this experiment and randomly divided into four groups: Sham group, I / R group, Bunazosin+I / R group and Bunazosin+Sham group.

[0032] (1) Sham group: except for not performing mesenteric clamping to block blood supply and removing the arterial clamp for reperfusion, all other operation steps of the mice were consistent with I / R;

[0033] (2) I / R group: intestinal I / R model was established in mice;

[0034] (3) Bunazosin hydrochloride solution + intestinal ischemia-reperfusion injury model group (Bunazosin+I / R): Mice were given 10 mg / kg bunazosin hydrochloride solution orally in advance, and the intestinal I / R model was established in mice 60 minutes later;

[0035] (4) Bunazosin hydrochloride solution + intestinal ischemia-reperfusion injury model group (Bunazosin+I / R): Mice were given 10 mg / kg bunazosin hydrochloride solution by gavage in advance, and the intestinal I / R model was established in the mice 60 minutes later.

[0036] 3. Detection indicators and technical methods:

[0037] (1) Observe and record the survival status of mice for 7 days.

[0038] (2) Evaluation of intestinal tissue damage: a. Hematoxylin and eosin (H&E) stained small intestinal segments, observed the morphological changes of small intestinal tissue under a light microscope and took photos. A pathologist who was unaware of the experimental grouping evaluated the tissue damage according to the intestinal injury scoring standard. b. Intestinal barrier permeability detection: 1) Fluorescein isothiocyanate-labeled dextran (FITC-dextran) was used to detect intestinal permeability: mice were gavaged at a dose of 0.6 mg / kg 4 hours before the test, and the FITC content in peripheral blood was detected using an enzyme marker (485 nm, 530 nm); 2) Detection of endotoxin (LPS) levels in plasma: Detection was performed according to the steps provided in the ELISA kit instructions. c. RT-PCR was used to detect the mRNA expression level of inflammatory factors in intestinal tissue.

[0039] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present invention, which should be covered by the scope of the claims of the present invention.

Claims

1. Use of bunazosin hydrochloride in the preparation of a medicament for treating intestinal ischemia-reperfusion injury, characterized in that: The bunazosin hydrochloride treats intestinal ischemia-reperfusion injury by alleviating intestinal ischemia-reperfusion injury, inhibiting damage to extraintestinal organs, increasing the survival time of model animals, inhibiting ischemia-reperfusion-induced oxidative stress and apoptosis, and promoting epithelial cell proliferation.

2. The use according to claim 1, characterized in that: The drug also includes pharmaceutically acceptable excipients.

3. The use according to claim 2, characterized in that The dosage form of the drug is injection, capsule, tablet, granule, suspension, emulsion, spray, powder, liposome, oral solution or pill.

4. The use according to claim 1, characterized in that The drug can be administered by intramuscular injection, subcutaneous injection, intravenous injection, oral administration, sublingual administration or spray administration.

Images