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Application of S-propargyl-cysteine serving as hydrogen sulfide donor in drug preparation

A technology of hydrogen sulfide donor and cysteine, which is applied in the direction of pharmaceutical formulations, drug combinations, medical preparations containing active ingredients, etc.

Inactive Publication Date: 2017-10-24
THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report about the effects of H2S on anti-inflammation, anti-fibrosis, and inhibition of lesion growth and adhesion formation in endometriosis models

Method used

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  • Application of S-propargyl-cysteine serving as hydrogen sulfide donor in drug preparation
  • Application of S-propargyl-cysteine serving as hydrogen sulfide donor in drug preparation
  • Application of S-propargyl-cysteine serving as hydrogen sulfide donor in drug preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 实施例1 炔丙基半胱氨酸抗IL-1β诱导的子宫内膜间质细胞炎症实验

[0036] 1.细胞活力测定,按照Dojindo的CCK8说明书方法进行;取对数生长期的细胞,胰酶消化后吹打成单细胞悬液;按照5000个细胞 / 孔的浓度接种至96孔板;待细胞贴壁(隔夜)稳定后,加入不含血清的不同浓度的SPRC(0.1-100μM)继续培养;选取6h、12h、24h、36h、48h时间点,按照每孔培养基总体积的10%加入CCK-8溶液(注意孔中不要产生气泡,以免影响OD值的读数)。 同时在不含细胞的培养基中加入CCK8即为空白对照。 37°C 5% CO 2 培养箱中孵育2h后酶标仪检测450nm波长的吸光度;

[0037] 2.流式检测凋亡,按照BD的ANNEXIN V-FITC细胞凋亡检测试剂盒说明书方法进行;取对数生长期的细胞,胰酶消化后吹打成单细胞悬液,接种至6孔板中;待细胞达到80%丰度时,加入不含血清的不同浓度的SPRC(1-10μM)继续培养24h;把细胞培养液吸出至合适离心管内,加入适量胰酶细胞消化液消化细胞。孵育至轻轻吹打可以使贴壁细胞吹打下来时,加入收集的细胞培养液,将细胞转移到离心管内,1000rpm,4℃离心5分钟;弃上清,加入预冷的PBS轻柔清洗细胞2次,将细胞重悬于Binding Buffer中,调整细胞的浓度为1×10 6 个 / ml;取100ul细胞悬液,加入5μl Annexin V-FITC和5μl PI,轻轻混匀,避光室温反应15分钟;设置阴性对照(不加Annexin V-FITC和PI)、单阳1(不加Annexin V-FITC,只加PI)、单阳2(不加PI,只加Annexin V-FITC)三个对照组;加入200ul Binding Buffer,在半小时内上机检测。注意事项:旋帽离心管装的试剂在开盖前请短暂离心,将盖内壁上的液体甩至管底,避免开盖时液体洒落。Annexin V-FITC和Propidium iodide是光敏物质,在操作时请注意避光。在细胞洗涤时请尽量将上清弃净,以免PBS残留,有可能会影响实验结果。整个操作过程动作要尽量轻柔,勿用力吹打细胞。反应完毕后请尽快检测,因为细胞凋亡是一个动态的过程,反应1小时后荧光强度就开始衰变;

[0038] 3.ELISA检测IL-6的分泌,3....

Embodiment 2

[0049] 实施例2 炔丙基半胱氨酸减轻子宫内膜异位症小鼠模型中内异症及其粘连

[0050] 1.自体子宫内膜缝合法小鼠模型

[0051] 1.1模型的建立

[0052] 用1%戊巴比妥钠(0.1ml / 20g)腹腔注射麻醉,待小鼠麻醉后,胶带固定四肢,仰卧位固定在鼠板上。剃去腹部鼠毛,碘酒消毒,75%乙醇脱碘。下腹部正中切口1cm,打开腹腔,暴露双侧子宫。分离一侧子宫,上下端结扎,游离出中间段约1-1.2cm,沿上下端剪下置于无菌生理盐水中,轻轻漂洗,除去脂肪组织和浆膜面,将宫腔沿纵轴剖开,平均分成3-4块,每块约3*3mm大小,7-0号带针缝线将子宫小段缝合到小鼠腹壁(内膜面和腹壁贴合)四个象限腹膜上。切口缝合、消毒。术中给予青霉素钠肌注(16000U / 只)预防感染,后置于无菌鼠笼中。假手术组除不缝合子宫内膜外,其余操作相同。标 准啮齿类动物饲料喂养,室温22-24℃,相对湿度50—60%,常规饮水及照明。

[0053] 1.2分组、检测、给药及取材

[0054]Female C57 mice, 20-22g. Behavioral testing was carried out before modeling, and 0.1 mg / kg of estradiol was given after the testing, and injected intramuscularly to improve the success rate of modeling. Modeling was established within 36-48 hours after intramuscular injection of estradiol. The model was established for 2 weeks, and estradiol was injected twice a week. Behavioral testing was carried out after 2 weeks and they were randomly divided into the following 5 groups: (1) SPRC model group (n=10), given normal saline, orally...

Embodiment 3

[0077] Example 3 The effect of propargyl cysteine ​​in uterine horn adhesion model rats

[0078] 1. Establishment of rat uterine horn adhesion model

[0079] Rats were anesthetized by intraperitoneal injection of 7% chloral hydrate (0.4ml / 100g), fixed on the rat board in the supine position, the rat hairs on the lower abdomen were shaved, and disinfected with iodine. Make a midline incision of about 3 cm in the lower abdomen of the rat, first use a razor blade to injure the bilateral abdominal wall, and acutely separate the SD rat cardiomyocytes: maintain a uniform force and scrape up and down 10 times, it is advisable to have slight bleeding, and burn the bilateral uterine horns with a 380-degree electric iron At 9-10 points, the uterine horns at both ends of the injury were sutured to the abdominal wall, one stitch at the upper and lower sides, with an interval of about 1 cm, the abdomen was closed in double layers, and the abdomen was opened for evaluation 14 days later. I...

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PUM

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Abstract

The invention belongs to the field of drug preparation, and relates to application of S-propargyl-cysteine (SPRC) serving as a hydrogen sulfide donor in the drug preparation. The S-propargyl-cysteine (SPRC) serving as the hydrogen sulfide donor is used for performing an experiment that H2S works on endometriosis and adhesion thereof through an endometrial stromal cell line, a C57 mouse endometriosis model and a Wistar rat uterine adhesion model. A result shows that the S-propargyl-cysteine (SPRC) serving as the hydrogen sulfide donor can effectively inhibit IL-1beta induced endometrial mesenchymal cell inflammation; thermal hyperalgesia of an endometriosis model mouse can be obviously improved, so that lesion is reduced, fibrosis is relieved, and formation of postoperative adhesion of a uterine adhesion model rat can be inhibited; and the S-propargyl-cysteine (SPRC) serving as the hydrogen sulfide donor can be further used for preparing a drug for treating the endometriosis and complications thereof.

Description

technical field [0001] The invention belongs to the field of medicine and pharmacy, and relates to the use of hydrogen sulfide donor propargyl cysteine ​​(SPRC) in pharmacy, in particular to the use of hydrogen sulfide donor propargyl cysteine ​​(SPRC) in the preparation and treatment of endometrium Atopy and its use in adhesion medicine. Background technique [0002] Endometriosis (EMs) is a common disease in women of childbearing age, and its incidence is as high as 10-15% in women of childbearing age. The main clinical manifestations are pelvic pain and infertility, which seriously affect the quality of life of women. The pathogenesis of the disease has not yet been clarified, and the clinical treatment is facing great difficulties and challenges. The current treatment to remove the lesion and anti-estrogen drug treatment can help improve the condition and relieve symptoms to a certain extent, but it is still difficult to avoid recurrence. relapse. Among them, pelvic-ab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/198A61P15/00
CPCA61K31/198
Inventor 朱依谆夏叶徐丛剑焦杨吴志勇丛青王莹
Owner THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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