CN114917230B - Cb-839在制备抑制角膜新生血管生成的药物中的应用 - Google Patents
Cb-839在制备抑制角膜新生血管生成的药物中的应用 Download PDFInfo
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Abstract
本发明属于眼部药品制备技术领域。针对目前角膜新生血管的预防与治疗手段会引发并发症的问题,本发明提供CB‑839在制备抑制角膜新生血管生成的药物中的应用。本发明通过负载CB‑839的药物能够明显抑制小鼠角膜新生血管生成,炎症细胞浸润减少,并且没有发现明显副作用,弥补了现有技术的不足。
Description
技术领域
本发明属于眼部药品制备技术领域,具体涉及CB-839在制备抑制角膜新生血管生成的药物中的应用。
背景技术
角膜新生血管的预防与治疗是一项重大的临床挑战。当角膜在遭受损伤、感染、角膜手术和遗传疾病等内外因素影响下,角膜中会出现过度氧化应激反应,引发促血管生成因子上调,进而引起毛细血管由角膜缘向角膜内侵入形成角膜新生血管,造成患者视力下降,严重时致盲。
目前临床上的药物和手术防治手段常伴随着青光眼、白内障、角膜上皮损伤或角膜重复感染等并发症,因此,研究新的技术来预防角膜新生血管是紧迫且必须的。
发明内容
针对目前角膜新生血管的预防与治疗手段会引发并发症的上述问题,本发明提供CB-839在制备抑制角膜新生血管生成的药物中的应用,通过负载CB-839的药物能够明显抑制小鼠角膜新生血管生成,并且没有发现并发症,弥补了现有技术的不足。
本发明是通过以下技术方案实现的:
CB-839在制备抑制角膜新生血管生成的药物中的应用。
所述的CB-839为非竞争性的谷氨酰胺酶1(glutaminase 1(GLS1)),是一种非竞争性的谷氨酰胺酶1选择性抑制剂,在三阴性乳腺癌、肺腺癌、软骨肉瘤和淋巴瘤等癌细胞系表现出显着的抗增殖活性。
进一步的,所述膜新生血管是角膜出现过度氧化应激反应,引发促血管生成因子上调,进而引起毛细血管由角膜缘向角膜内侵入形成的角膜新生血管。
进一步的,所述的抑制角膜新生血管生成的药物剂型为缓释水凝胶、脂质体或聚乳酸-羟基乙酸共聚物微球。
进一步的,所述缓释剂为水凝胶缓释剂,其制备方法包括如下步骤:明胶与CB-839混合均匀制成水溶液,往制备的水溶液中加入羧甲基纤维素和N-羟基磺基琥珀酰亚胺钠盐,混合均匀,室温静置,即得负载CB-839的水凝胶缓释材料。
进一步的,明胶与CB-839混合均匀制成的水溶液中,明胶的浓度为0.03-0.08mg/mL,CB-839的浓度为0.3-0.5mg/mL。
进一步的,明胶与CB-839混合均匀制成的水溶液中,明胶的浓度为0.05mg/mL,CB-839的浓度为0.4mg/mL。
进一步的,所述缓释剂为脂质体缓释剂,其制备方法包括如下步骤:将大豆卵磷脂、胆固醇和CB-839中共同分散于二氯甲烷中形成均匀溶液,将溶液减压蒸干,然后分散于水中,离心,移去上清液,将沉淀物再加入水混匀,得到负载CB-839的脂质体缓释材料。
进一步的,所述分散为超声分散。
本发明的有益效果:能够有效抑制角膜新生血管生成;能够用于角膜新生血管生成持续治疗;解决了CB-839溶解度差和生物利用度低的缺点,可以减少多次和频繁使用传统制剂的不便性,使药物达到高效而持久的治疗效果。
附图说明
图1为动物实验小鼠眼部大体照。
具体实施方式
下面结合具体实施例对本发明做进一步详细说明。
实施例1:制备负载CB-839的水凝胶缓释材料
(1)向离心管中加入50mg明胶和2mg CB-839;
(2)向步骤(1)的离心管中加入1mL水;
(3)将上述混合体系在常温下超声20min使其完全溶解,形成均匀溶液;
(4)向步骤(3)的离心管中加入4.5mg的羧甲基纤维素和1.8mg N-羟基磺基琥珀酰亚胺钠盐的催化剂;
(5)将步骤(4)中所得到的盛有混合溶液的离心管室温放置1h,既得负载CB-839的水凝胶缓释材料。
实施例2:制备负载CB-839的脂质体缓释材料
(1)向圆底烧瓶中加入60mg大豆卵磷脂、16mg胆固醇和4mg CB-839;
(2)向步骤(1)的圆底烧瓶中加入20mL二氯甲烷;
(3)将上述混合体系在常温下超声20min使其完全溶解,形成均匀溶液;
(4)将(3)中溶液减压缓慢蒸干;
(5)将步骤(4)中圆底烧瓶中加入20mL水,水合1小时;
(6)将步骤(5)中混合液超声2min;
(7)将步骤(6)中混合液以6000转/分钟离心5min,移去上清液;
(8)向步骤(7)中沉淀物中加入10mL水,混匀,既得负载CB-839的脂质体缓释材料。
动物实验
①碱烧伤动物模型建立
(1)将直径约2毫米的圆形滤纸浸泡在1mol/L NaOH溶液中。
(2)90mg/kg戊巴比妥钠注射麻醉Balb/c小鼠,每25g小鼠注射戊巴比妥钠0.3mL,相当于每25g小鼠注射戊巴比妥钠1.8mg。麻醉应该需要1-2分钟才能开始。麻醉深度可以通过轻轻捏动物的脚趾或尾巴来确定,如果麻醉足够,应该没有反应。
(3)将0.5%盐酸丙美卡因滴在小鼠角膜表面局部镇痛。
(4)用无菌钳夹起一张NaOH浸泡过的滤纸。如果过量的氢氧化钠附着在滤纸上或从滤纸上滴落下来,用干滤纸轻拍浸泡过的滤纸以吸收多余的氢氧化钠。将浸泡过氢氧化钠的滤纸放在角膜中央。放置40秒,可产生面积约2x2 mm2的急性碱烧伤。
(5)取下滤纸。使用10mL注射器,用10mL0.9%生理盐水轻轻冲洗眼睛40秒,洗去残留的1mol/L NaOH。
(6)实验组小鼠在眼球结膜下轻轻剪开约1mm的切口,用无菌镊将约0.05mg的负载CB-839的水凝胶缓释材料(实施例1)植入结膜下,并用11-0马尼缝线缝合切口,对照组小鼠不作处理。
(7)使用眼科检查用裂隙灯,在5天拍摄具有代表性的眼部大体照。图1为实验小鼠眼部大体照。结果显示,对照组血管比较明显(图1A),而实验组新生血管不明显(图1C),证实该水凝胶缓释材料释放CB-839,抑制了角膜新生血管的生成。表现为实验组小鼠角膜新生血管面积降低、炎症细胞浸润减少(图1B和D),并未发现明显副作用。
以上所述的实施例仅是对本发明的优选实施方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案作出的各种变形和改进,均应落入本发明权利要求书确定的保护范围内。
Claims (8)
1.CB-839在制备抑制角膜新生血管生成的药物中的应用。
2.根据权利要求1所述的应用,其特征在于,所述膜新生血管是角膜出现过度氧化应激反应,引发促血管生成因子上调,进而引起毛细血管由角膜缘向角膜内侵入形成的角膜新生血管。
3.根据权利要求1所述的应用,其特征在于,所述的抑制角膜新生血管生成的药物剂型为水凝胶缓释剂、脂质体缓释剂或聚乳酸-羟基乙酸共聚物微球。
4.根据权利要求3所述的应用,其特征在于,所述水凝胶缓释剂的制备方法包括如下步骤:明胶与CB-839混合均匀制成水溶液,往制备的水溶液中加入羧甲基纤维素和N-羟基磺基琥珀酰亚胺钠盐,混合均匀,室温静置,即得负载CB-839的水凝胶缓释剂。
5.根据权利要求4所述的应用,其特征在于,明胶与CB-839混合均匀制成的水溶液中,明胶的浓度为0.03-0.08mg/mL,CB-839的浓度为0.3-0.5mg/mL。
6.根据权利要求5所述的应用,其特征在于,明胶与CB-839混合均匀制成的水溶液中,明胶的浓度为0.05mg/mL,CB-839的浓度为0.4mg/mL。
7.根据权利要求3所述的应用,其特征在于,所述脂质体缓释剂的制备方法包括如下步骤:将大豆卵磷脂、胆固醇和CB-839中共同分散于二氯甲烷中形成均匀溶液,将溶液减压蒸干,然后分散于水中,离心,移去上清液,将沉淀物再加入水混匀,得到负载CB-839的脂质体缓释剂。
8.根据权利要求7所述的应用,其特征在于,所述分散为超声分散。
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