CN113769070A - 具有触变性和酸敏感性的胃粘膜创面保护胶及其应用 - Google Patents
具有触变性和酸敏感性的胃粘膜创面保护胶及其应用 Download PDFInfo
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- CN113769070A CN113769070A CN202110882542.6A CN202110882542A CN113769070A CN 113769070 A CN113769070 A CN 113769070A CN 202110882542 A CN202110882542 A CN 202110882542A CN 113769070 A CN113769070 A CN 113769070A
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Abstract
本发明提供了一种具有触变性和酸敏感性的胃粘膜创面保护胶及其应用，属于生物医药制剂领域。具有触变性的胃粘膜创面保护胶有效成分为增粘剂、酸敏感成胶物质、触变剂、增塑剂、缓凝剂、酸碱度调节剂，所述胃损伤粘膜创面保护胶的pH值≥7。还可以包括固化液，将保护胶在指定位置固化。该保护胶可在溃疡表面形成胶体膜，促进溃疡的愈合。此外本发明使产品具有触变性在创面上短时间内保持不流动状态，然后再采用固化液使得保护胶迅速固定在损伤粘膜表面，解决了保护胶的流动问题。动物试验表明本发明能明显促进损伤性胃粘膜的修复，因此对胃粘膜损伤具有促进愈合和保护作用。
Description
技术领域
本发明涉及生物医药技术领域，特别是涉及一种适用于通过消化道内窥镜喷涂到消化道损伤粘膜表面的具有触变性和酸敏感性的胃粘膜创面保护胶及其制备方法和应用。
背景技术
内窥镜下胃粘膜切除手术和胃粘膜剥离术是早期胃癌和胃良性肿瘤切除的微创手术，不仅可达到根治早期胃癌的目的，且具有创伤小、对患者生存质量影响小等优点，已逐渐取代部分传统外科手术。食道和结肠的早期癌变和良性肿瘤切除的微创手术也取代部分的传统外科手术。但是，目前这种手术后创面仅做止血处理，尚无良好的创面保护措施。裸露的创面在消化液的作用下经历溃疡活动期、愈合期和瘢痕期，并且创面的愈合时间也较长。临床研究资料显示病人经内窥镜下胃粘膜切除手术和胃粘膜剥离术6周的愈合率为69％左右。因此，有必要提供一种可以直接通过胃镜应用到胃损伤粘膜表面的产品。
海藻酸盐作为优良的药物制剂基质，医药上多用作亲水性乳化剂、凝胶剂和增稠剂。中国专利[申请号：101933894B]公开了一种胃粘膜保护胶，包括成胶物质，酸碱调节剂、交联剂和粘合剂；所述成胶物质，交联剂和粘合剂重量比为1∶ 0.01-0.3∶0.1-0.5，酸碱调节剂的用量为调pH在6.5-8.5。该专利仅利用纯物理学原理保护胃黏膜，使用的交联剂为酸溶解性钙，这种钙在中性溶液中不溶解呈颗粒状态，适合口服使用，不适合通过注射器注射。此外产品采用的粘附剂为阴离子聚合物和中性聚合物，而不是粘合剂，胃损伤粘膜表面也是带阴离子，因此不能很好的粘合到胃粘膜的创面上，非常容易脱落，不能充分发挥胃粘膜的保护作用。
发明内容
针对上述技术领域中的不足，本发明提供一种全新的采用物理原理并且具有粘合力的保护胃粘膜创面的方法和产品，该产品具有触变性，受注射应力作用后粘度明显减低具有可注射性，注射应力撤销后粘度明显提高流动性减低，可以避免在创面的瞬间流动，在消化道内酸性环境条件以及不需要钙离子的情况下高分子聚合物和高分子聚合物之间进一步交联形成胶体膜，起到保护消化道粘膜的作用。
本发明提供一种具有触变性和酸敏感的胃粘膜创面保护胶，包括胶体部分和固定液部分，其胶体液部分有效成分为增粘剂、酸敏感成胶物质、触变剂、促进细胞迁移材料、增塑剂、缓凝剂、酸碱度调节剂，所述胃粘膜粘膜创面保护胶的pH值≥ 7。
所述的增粘剂为羧甲基壳聚糖、多聚赖氨酸、三聚赖氨酸、贻贝粘性蛋白、胶原蛋白、聚多巴胺、壳聚糖季铵盐。
所述酸敏感成胶物质为果胶、海藻酸盐、葡甘聚糖、卡拉胶、黄原胶、瓜尔豆胶中的两种或两种以上。
所述果胶为低甲氧基果胶和酰胺化低甲氧基果胶中的一种或两种，海藻酸盐为海藻酸钠、海藻酸钾的一种或两种。
所述保护胶中还包括酸碱调节剂，酸碱调节剂用于调节保护胶的pH值≥7。
所述保护胶中成胶物质和增粘剂的重量比是3:1-3。
所述的促进细胞迁移材料为甘草多糖、β-葡聚糖、党参多糖、索拉胶、杏鲍菇葡聚糖、燕麦葡聚糖、灰树花多糖、谷物葡聚糖、酵母葡聚糖、灵芝多糖、胶原蛋白降解产物、肝细胞生长因子、表皮细胞生长因子、胰岛素样生长因子1、成纤维细胞生长因子、血小板来源生长因子、富组蛋白1、角质细胞生长因子、抗菌肽中的一种或多种。用量为0.001％-0.5％(W/V)。
所述的触变剂为硅酸镁锂，用量为1.0％-5％(W/V)。
所述酸碱调节剂为氢氧化钠、氢氧化钾、碳酸氢钠、碳酸氢钾、磷酸钠、磷酸氢二钠、乙酸钠、乳酸钠的一种或多种。
所述的增塑剂为甘油、聚乙二醇、聚乙烯醇、甘露醇和山梨醇的一种或多种。
所述的缓凝剂为葡萄糖酸钠、柠檬酸钠、酒石酸钠、水杨酸钠、焦磷酸钠、二聚磷酸钠、三聚磷酸钠、磷酸二氢钾、磷酸二氢钠、醋酸钠、四硼酸钠、乙二胺四乙酸二钠的一种。用量0％-1％。
所述的固定液为钙盐溶液。钙盐为氯化钙、葡萄酸钙、乳酸钙的一种或多种。
本发明还提供一种前述的具有触变性的胃粘膜创面保护胶在治疗消化性溃疡、应激性溃疡、胃炎、胃出血、胃粘膜切除和剥离术、食道粘膜剥离术和十二指肠道粘膜剥离术后的创面药物中的应用。
由于消化道基本处于酸性或弱酸性环境，因此本发明的目的均能实现。本发明在描述效果时，主要以胃部环境进行描写，但是并不代表本发明仅局限于胃部，本发明可以应用于人体消化道，包括食道下端，胃，十二指肠上端等。
与现有技术相比，本发明具有以下优点：
本发明采用科学原理，在胃酸条件下，使成胶物质在胃肠粘膜表面形成胶体膜，阻断胃酸和胃蛋白酶对粘膜的消化作用,促进溃疡愈合。
具有触变性的胃粘膜创面保护胶产品在使用前，溶液状态下为中性或偏碱性质，产品不形成凝胶状态。产品具有触变性和可注射性避免了注射后的瞬间流动，而在酸性条件下，使本品成凝胶膜，而增粘剂提供粘结于胃粘膜的性能，使凝胶膜附着于胃粘膜上，阻断胃酸和胃蛋白酶对粘膜的消化作用，起到保护胃粘膜的作用。
本发明基于酸敏感成胶物质海藻酸盐、葡甘聚糖、黄原胶等材料的凝胶机理，在偏碱性环境条件下，分子之间不能交联为液体状态。和胃酸接触作用后，在酸性条件下海藻酸盐、葡甘聚糖、增粘剂等材料发生分子间交联而形成凝胶膜。
具有触变性的胃粘膜创面保护胶是采用从植物提取的天然高分子材料海藻酸钠、黄原胶、葡甘聚糖等以及增粘剂制成，通过胃窥镜喷洒在胃损伤粘膜表面形成胶体膜，起到保护胃粘膜的作用。本产品的特点是在通常情况下为液体，在胃酸的作用下，保护胶中的分子相互交联，在胃损伤粘膜表面形成胶体膜，避免胃酸和胃蛋白酶对胃肠粘膜的消化作用，促进溃疡的愈合。
本发明增加了促进细胞迁移材料，能促进创面粘膜细胞增殖和迁移能力，具有促进创面愈合作用。
由于粘膜创面保护胶涂到损伤粘膜后不会马上和胃酸接触，会有流动的问题，所以本发明选择合适的生物安全无毒性的触变剂使得产品具有触变性和可注射性，在创面上短时间内保持不流动状态，然后再采用固化液使得保护胶迅速固定在损伤粘膜表面，解决了胃粘膜创面保护胶的流动问题。
申请人之前申请过消化道创面保护胶专利，但是其中胶体液部分需要有钙离子和钙离子络合剂，在胃内酸性条件下络合的钙离子释放出来使得成胶物质发生交联成膜。由于胃壁创面有的不是处在水平面状态，液体的流动性在注射部位会发生瞬间的流动，因此本发明引入了触变剂，使其具有可注射性而降低其流动性，即是处在垂直状态或天花板状态的胃壁粘膜创面也不会发生明显流动；添加缓凝剂使形成的胶体脆性降低并提高粘附性。因此本发明不再使用钙离子的情况下实现了在酸性条件下的成胶物质的交联成膜，形成的胃粘膜保护胶具有更好的使用性能。
具体实施方式
下面结合实施例对本发明作进一步的详细说明。
本发明实施例中没有特殊说明，采用的均为一般化学品，市场上购买即可得到。
实施例1
1.海藻酸钾12克
2.葡甘聚糖2克
3.多聚赖氨酸12g
4.β-葡聚糖0.2g
5.甘油10g
6.醋酸钠3g
组分1：取硅酸镁锂7.5克溶解在采用1mol氢氧化钠调pH8.0的生理盐水500ml中混匀，然后将上述成分加入搅拌至完全溶解，每瓶5ml包装,高温蒸汽灭菌。
组分2：固化液采用氯化钙配制成1.1％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例2
1.酰胺化低甲氧基果胶6g
2.葡甘聚糖2g
3海藻酸钾8g
4.壳聚糖12g
5.燕麦葡聚糖0.2g
6.甘油10g
7.葡萄酸钠1g
组分1：取硅酸镁锂7.5克溶解在采用碳酸氢钠调pH8.0的生理盐水500ml中混匀，然后将上述成分加入搅拌至完全溶解，每瓶5ml包装,高温蒸汽灭菌。
组分2：固化液采用氯化钙配制成1.1％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例3
1.海藻酸钠12g
2.葡甘聚糖2.5g
3.多聚赖氨酸4g
4.燕麦葡聚糖0.2g
5.聚乙烯醇2.5g
6.柠檬酸钠5g
组分1：取硅酸镁锂7.5克溶解在采用1mol氢氧化钠调pH8.0的生理盐水500ml中混匀，然后将上述成分加入搅拌至完全溶解，每瓶5ml包装,高温蒸汽灭菌。
组分2：固化液采用葡萄糖酸钙配制成2％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例4
1.海藻酸钠10g
2.黄原胶2.5g
4.多聚赖氨酸10g
5.灰树花多糖0.2g
5.聚乙二醇5g
6.酒石酸钠1.5g
组分1：取硅酸镁锂7.5克溶解在采用碳酸氢钠调pH8.0的生理盐水500ml中混匀，然后将上述成分加入搅拌至完全溶解，每瓶5ml包装,高温蒸汽灭菌。
组分2：固化液采用氯化钙配制成1.7％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实施例5
1.海藻酸钾10克
3.黄原胶2g
4.多聚赖氨酸10g
5.谷物葡聚糖0.2g
5.聚乙烯醇2.5g
组分1：取硅酸镁锂7.5克溶解在采用1mol氢氧化钠调pH8.0的生理盐水500ml中混匀，然后将上述成分加入搅拌至完全溶解，每瓶5ml包装,高温蒸汽灭菌。
组分2：固化液采用乳酸钙配制成5％的水溶液，每瓶5ml包装,高温蒸汽灭菌。
实验例效果评价
以上实施例的液体的直接用于试验。
1体外胶体膜形成试验
分别取上述液体2ml涂布在新鲜的猪皮内侧面上，然后再喷洒固化液，观察胶体膜形成的情况。结果是实施例1到实施例5均形成胶体膜。
分别取上述液体2ml涂布在新鲜的猪皮内侧面上，然后再喷洒人工胃液，观察胶体膜形成的情况。结果是实施例1到实施例5均形成胶体膜。
2触变性测试：
用博力飞DV－Ⅱ型粘度计进行粘度测定。并按照公式(1)计算假塑比值进行比较，选用50rpm和200rpm转速下的粘度进行计算。以羧甲基纤维素溶液作为参照。
假塑比值＝高转速下的粘度/低转速下的粘度(1)。
结果：2％羧甲基纤维素溶液假塑比值0.626，样品1假塑比值0.233；样品2假塑比值0.246；样品3假塑比值0.236；样品4假塑比值0.266，样品5假塑比值0.248。
3安全性研究试验
将实施例1-5进行以下生物学试验
1)口腔粘膜刺激：将12ml胶体液倒入直径15cm的无菌培养皿中，然后倒入固定液后30分钟，用生理盐水冲吸收掉多余的固定液，然后按照3cm2/ml比例加入生理盐水，在37摄氏度浸提72小时制备试验液。将样品的实验液制成直径不大于5mm棉球浸透置于3只金黄地鼠一侧颊囊中。接触时间每次最少5min，每天一次，共4次，末次接触后24h肉眼观察颊囊,无痛处死地鼠,取颊囊有代表性部位的组织样品放入4％甲醛溶液中固定后并制作组织切片后进行组织学评价。结果刺激指数均为0，被试样品无口腔粘膜刺激性。
2)细胞毒性：将12ml胶体液倒入直径15cm的无菌培养皿中，然后倒入固定液后30分钟，用生理盐水冲吸收掉多余的固定液，然后按照3cm2/ml比例加入培养基，在37摄氏度浸提24小时制备试验液。然后按GB/T16886.5中规定的细胞毒性试验采用 MTT法测定，细胞增殖率在95％-105％范围内。
3)致敏试验：将12ml胶体液倒入直径15cm的无菌培养皿中，然后倒入固定液后 30分钟，用生理盐水冲吸收掉多余的固定液，然后按照3cm2/ml比例加入生理盐水，在37摄氏度浸提72小时制备试验液。然后按GB/T16886.10中规定的方法进行皮肤致敏试验，均未观察到致敏作用。
4组织粘附力的测定(组织留存量法)：
将实施例1-5分别进行以下试验
取SD大鼠,禁食24h用戊巴比妥钠溶液(40mg/kg)经腹腔注射麻醉,解剖取出胃置生理盐水(37℃)中切取胃,用生理盐水将胃内壁清洗干净，清洗的胃在2小时内使用。剪取一定面积的胃组织(2cm×2cm),固定于聚乙烯薄膜上，将0.5ml保护胶均匀涂布于胃黏膜创面上，然后喷涂固化液。胃组织在相对湿度为92.5％的恒湿密闭容器中放置20分钟，将经上述处理的胃组织固定于冲洗斜槽上,将斜槽的角度调至60度，调节蠕动泵流速为20ml/min,将胃组织用0.1mol/L盐酸冲洗5mins,冲洗液收集于一已知重量的烧杯中,在70℃烘干,称重,组织粘附力用粘附百分率表示。
计算方法如下:
胃组织粘附百分率(Bg/％)Bg/％＝{[M-(G-g-m)]/M}x100％
式中M为黏膜保护胶的重量(0.5ml同条件下烘干)；g为空烧杯重；G为烧杯与烘干后残渣总重；m为同体积冲洗液所含固体物质的量(空白对照)。B值越大表示粘附力越大。
结果显示：实施例1：98％，实施2：95％，实施例3：97％，实施例4：98％，实施例5：97％。试验表明实施例1-5的双组分自粘性胃黏膜保护胶对组织的粘附力较强，不易脱落。
5.动物试验
动物分两组，体重3kg左右。实验组6只，对照组6只。
手术前24小时禁食，不禁水。
麻醉方法：推荐家兔用质量浓度30g/L戊巴比妥钠静脉注射1.0ml/kg。
家兔仰面固定手术台上，腹部去毛。按外科常规手术要求以2％碘酊和75％乙醇溶液消毒试验区域。
在上腹部逐层切开皮肤、肌肉层和腹膜，如有出血结扎止血。暴露胃部，在胃大弯处切开胃，用生理盐水清洗胃部，在胃体内的胃大弯侧面的黏膜下，1：1000 0肾上腺素生理盐水进行黏膜下注射，注射1ml生理盐水形成胃黏膜突出，然后用环套器切除直径1cm的黏膜形成创面，喷洒凝血酶(1ml内含凝血酶50U)并压迫止血完全，测量创面的直径。对照组不做任何处理，实验组涂布0.5ml保护胶，再喷涂固化液后形成固体化的保护膜，然后缝合胃部。再逐层缝合腹壁。放饲养笼中，禁食一天。
结果观察：与手术后1周安乐死动物，沿原来的切口切开胃壁，观察创面愈合情况，测量创面的直径，结果发现实验组手术后1周溃疡的实验组6只家兔愈合，愈合率100％；对照组3只愈合，3只未愈合，愈合率50％。以上结果可以看出,按实施例 1能明显促进的胃黏膜损伤性溃疡愈合，对胃溃疡有明显的治疗作用。
对比例1
以上成份卡波姆溶解在45ml纯水中，酸溶壳聚糖、羟丙基甲基纤维素、海藻酸钠和β-葡聚糖溶解在45ml醋酸溶液中，然后将两液体混合，用三乙醇胺调pH到5.5，然后定容到100ml，每瓶5ml包装,高温蒸汽灭菌。
固化液采用氯化钙配制成1.1％的水溶液，调pH＝8.0，每瓶5ml包装,高温蒸汽灭菌。
对比例2
以上成份卡波姆溶解在45ml生理水中，酸溶胶原蛋白、海藻酸钠和β-葡聚糖溶解45ml醋酸溶液中，然后将两液体混合，用三乙醇胺调pH到5.0，然后定容到 100ml，每瓶5ml包装,高温蒸汽灭菌。
固化液采用氯化钙配制成1.1％的水溶液，调pH＝8.0，每瓶5ml包装,高温蒸汽灭菌。
对比例3
以上成份卡波姆溶解在45ml生理水中，酸溶壳聚糖、胶原蛋白、羟丙基甲基纤维素、海藻酸钠和β-葡聚糖溶解在45ml醋酸溶液中，然后将两液体混合，用氢氧化钠溶液调pH到5.5，然后定容到100ml，每瓶5ml包装,高温蒸汽灭菌。
固化液采用葡萄糖酸钙配制成2％的水溶液，调pH＝8.0，每瓶5ml包装,高温蒸汽灭菌。
实验例效果评价
1体外胶体膜形成试验
分别取上述液体2ml涂布在新鲜的猪皮内侧面上，然后再喷洒固化液，观察胶体膜形成的情况。结果是实施例1到实施例5均形成胶体膜。对比例1到实施例3形成固定膜。
分别取上述液体2ml涂布在新鲜的猪皮内侧面上，然后再喷洒人工胃液，观察胶体膜形成的情况。现在申请专利的实施例1到实施例5均形成胶体膜。以前申请的专利对比例1-3均不能成膜。
表1：实施例和对比例的成膜效果比较
| 固定液成膜 | 酸成膜性 | |
| 实施例1 | 能成膜 | 能成膜 |
| 实施例2 | 能成膜 | 能成膜 |
| 实施例3 | 能成膜 | 能成膜 |
| 实施例4 | 能成膜 | 能成膜 |
| 实施例5 | 能成膜 | 能成膜 |
| 对比例1 | 能成膜 | 不能成膜 |
| 对比例2 | 能成膜 | 不能成膜 |
| 对比例3 | 能成膜 | 不能成膜 |
从对比例与本发明实施例可以看到，本发明的保护胶具有酸成膜性，可以在胃部更好更膜。
本文中所描述的具体实施例仅仅是对本发明精神作举例说明。本发明所属技术领域的技术人员可以对所描述的具体实施例做各种各样的修改或补充或采用类似的方式替代，但并不会偏离本发明的精神。
Claims (11)
1.具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，所述保护胶包括胶体部分和固定液部分，其中胶体液部分有效成分为增粘剂、酸敏感成胶物质、触变剂、增塑剂、缓凝剂和酸碱度调节剂；所述胃粘膜粘膜创面保护胶的pH值≥7。
2.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，所述增粘剂为羧甲基壳聚糖、多聚赖氨酸、三聚赖氨酸、贻贝粘性蛋白、胶原蛋白、聚多巴胺、壳聚糖季铵盐中的一种或多种。
3.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，所述酸敏感成胶物质为果胶、海藻酸盐、葡甘聚糖、卡拉胶、黄原胶、瓜尔豆胶中的两种或两种以上。
4.根据权利要求3所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，所述果胶为低甲氧基果胶和酰胺化低甲氧基果胶中的一种或两种，海藻酸盐为海藻酸钠、海藻酸钾的一种或两种。
5.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，所述保护胶中酸敏感成胶物质和增粘剂的重量比是3:1-3。
6.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，保护胶还包括促进细胞迁移材料，促进细胞迁移材料为甘草多糖、β-葡聚糖、党参多糖、索拉胶、杏鲍菇葡聚糖、燕麦葡聚糖、灰树花多糖、谷物葡聚糖、酵母葡聚糖、灵芝多糖、胶原蛋白降解产物、肝细胞生长因子、表皮细胞生长因子、胰岛素样生长因子1、成纤维细胞生长因子、血小板来源生长因子、富组蛋白1、角质细胞生长因子、抗菌肽中的一种或多种。
7.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，触变剂为硅酸镁锂，用量为1.0％-5％(W/V)。
8.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，增塑剂为甘油、聚乙二醇、聚乙烯醇、甘露醇和山梨醇的一种或多种。
9.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，缓凝剂为葡萄糖酸钠、柠檬酸钠、酒石酸钠、水杨酸钠、焦磷酸钠、二聚磷酸钠、三聚磷酸钠、磷酸二氢钾、磷酸二氢钠、醋酸钠、四硼酸钠、乙二胺四乙酸二钠的一种或多种。
10.根据权利要求1所述的具有触变性和酸敏感性的胃粘膜创面保护胶，其特征在于，固定液为钙盐溶液，钙盐为氯化钙、葡萄酸钙、乳酸钙的一种或多种。
11.一种如权利要求1任意一项所述的具有触变性和酸敏感性的胃粘膜创面保护胶在制备治疗消化性溃疡、应激性溃疡、胃炎、胃出血、胃粘膜切除和剥离术、食道粘膜剥离术和肠道粘膜剥离术后的创面药物中的应用。
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