CN109675043A - 甲硫氨酸在增敏紫杉醇对乳腺癌治疗效果中的应用 - Google Patents
甲硫氨酸在增敏紫杉醇对乳腺癌治疗效果中的应用 Download PDFInfo
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Abstract
本发明公开了合用甲硫氨酸对增强紫杉醇治疗乳腺癌药效的应用。通过人源乳腺癌细胞MCF‑7小鼠模型,分别单独给予紫杉醇、甲硫氨酸以及两者联合给药,发现甲硫氨酸自身无显著抗肿瘤效应,但可显著提高紫杉醇对人乳腺癌细胞的抗肿瘤药效作用。在MCF‑7细胞模型中,在培养基中额外补充甲硫氨酸可显著增强紫杉醇对肿瘤细胞的杀伤作用。本发明所述甲硫氨酸可以用于合用以提高紫杉醇化疗疗效,或与紫杉醇共同制备复方增敏药物,以提高紫杉醇对乳腺癌细胞治疗的效果,为临床治疗乳腺癌提供了一个方便、经济、高效的增敏剂。
Description
技术领域
本发明属于肿瘤化疗的增敏药物领域,涉及甲硫氨酸的一种新用途,即甲硫氨酸用于增强紫杉醇治疗乳腺癌药效及其应用。
背景技术
作为严重危害女性身心健康的恶性肿瘤,乳腺癌的发病率在我国所有女性恶性肿瘤中居首位。目前的乳腺癌治疗中,化疗仍然是较为有效且被普遍使用的一种手段,即利用抗癌药物抑制癌细胞分裂,破坏癌细胞的治疗方法,化疗可以有效地降低患者死亡率并延长寿命。但是,化疗药物在临床应用中存在诸多问题,例如多药耐药现象、对正常细胞及组织的毒性等,严重影响了临床化疗的治愈率,据估计有相当一部分乳腺癌患者(高达70%)对于当前的化疗方案响应不佳。紫杉醇(Paclitaxel,PTX)是从裸子植物红豆杉的树皮分离提纯的天然次生代谢产物,经临床验证,具有良好的抗肿瘤作用。因其高效、广谱的特点,紫杉醇从分离至今已被广泛应用于多种恶性肿瘤,也是乳腺癌治疗的一线药物。但不可避免地,紫杉醇在临床使用中也存在很多缺陷,致使疗效降低甚至消失。化疗增敏药物是指自身没有(或很小)抗肿瘤作用,但能够显著增强抗肿瘤药物对肿瘤细胞杀伤作用的化合物(或药物)的总称。国内外研究人员一直致力于研发化疗增敏剂,期望能够提高化疗药物疗效从而延长患者的生命并提高患者生活质量。
甲硫氨酸(Methioine)为构成人体的必需氨基酸之一,可作为氨基酸类药物用于临床,可有效改善肝脏、心肌机能,其临床应用效果可靠且安全。目前还没有甲硫氨酸能有效地提高肿瘤化疗药物敏感性的报道。本发明人研究发现甲硫氨酸可显著增强紫杉醇的抗肿瘤效应。因此,发明人提出,甲硫氨酸与抗肿瘤药物联用可增强化疗药物敏感性,提高临床疗效。
发明内容
本发明以人源乳腺癌细胞MCF-7接种的荷瘤小鼠作为模型动物,通过腹腔注射给予甲硫氨酸联合紫杉醇治疗,测定小鼠肿瘤体积的变化评价甲硫氨酸对紫杉醇的增敏作用,发现甲硫氨酸单用组没有明显的抑瘤作用,紫杉醇单用组呈现出一定的抑瘤作用;与紫杉醇单用相比,紫杉醇与甲硫氨酸联用可显著增强紫杉醇的抗肿瘤药效。此外,本发明以MCF-7为细胞模型,测定细胞存活率评价甲硫氨酸对紫杉醇的增敏作用,发现在培养基中额外补充甲硫氨酸可显著增强紫杉醇对肿瘤细胞的抑制作用。以此为基础,本发明提出甲硫氨酸与紫杉醇联用可应用于乳腺癌的临床治疗。
附图说明
附图1 MCF-7荷瘤小鼠肿瘤大小与形态照片
附图2 MCF-7荷瘤小鼠肿瘤体积变化曲线
附图3 MCF-7荷瘤小鼠肿瘤重量对比图
附图4 MCF-7细胞存活率
附图1-3中,Control为对照组(生理盐水,腹腔注射),PTX为紫杉醇单用组(5mg/kg/every 3day,腹腔注射),Methionine为甲硫氨酸补充组(120mg/kg/day,腹腔注射),PTX+Methionine为紫杉醇联合甲硫氨酸组。*P<0.05,**P<0.01vs紫杉醇(PTX)单用组。
附图4中组别分别为:甲硫氨酸(30μg/mL)+PTX(0nM)、甲硫氨酸(30μg/mL)+PTX(10nM)、甲硫氨酸(75μg/mL)+PTX(10nM)、甲硫氨酸(150μg/mL)+PTX(10nM)。*P<0.05,**P<0.01vs甲硫氨酸(30μg/mL)+PTX(0nM)组;#P<0.05,##P<0.01vs甲硫氨酸(30μg/mL)+PTX(10nM)组。注:细胞培养基中甲硫氨酸本底含量即为30μg/mL。
具体实施方式
本发明通过下面的实施例进行详细的解释,但并不意味着本发明仅限于此。
细胞来源与培养:人乳腺癌细胞MCF-7购于中国医学科学院天津血液病研究所(天津,中国),细胞培养在37℃,含5%CO2的环境中,培养液为RPMI1640培养基,含10%胎牛血清(FBS),以及链霉素100mg/L,青霉素62.5mg/L。细胞生长于10cm培养皿中,2~3天更换培养基一次,待细胞生长至90%融合时进行传代。
动物来源与饲养:采用Balb/c-nu/nu小鼠(雌性,由南京大学模式动物研究所提供,周龄4~6,体重20±2g)进行实验。在标准饲养环境下(自由饮食与饮水,昼夜交替,各12小时),适应性饲养一周。动物饲养和全部实验于中国药科大学SPF级小鼠饲养室完成。
MCF-7荷瘤小鼠模型的建立:将MCF-7细胞消化,无菌PBS重悬后,在小鼠上肢腋下皮下注射入体内进行接种,每只小鼠接种106~107个/0.2mL,每日观察老鼠状态并测量瘤体积。1~2周后,选取出接种成功的荷瘤小鼠(n=5),待瘤体达100mm3后,将荷瘤小鼠按体重随机分为4组,开始按照需要给药处理。
MCF-7荷瘤小鼠实验设计和分组如下:Control组(生理盐水,腹腔注射),PTX组(5mg/kg/every 3day,腹腔注射),Methionine组(120mg/kg/day,腹腔注射),PTX+Methionine组。每隔两或三日测量小鼠体重及瘤体的长径和短径,计算瘤体积并绘制曲线。给药3周之后,处死动物,迅速取出肿瘤组织,清洗拭干后,称重并拍照。
CCK-8法测定MCF-7细胞活性实验设计和分组:取对数生长期的MCF-7细胞,用0.1%胰酶消化细胞,以5000cell/孔接种于96孔培养板中。24小时后,待细胞基本贴壁后更换为含药培养基。给药方案如下:甲硫氨酸(30μg/mL)+PTX(0nM)(对照组)、甲硫氨酸(30μg/mL)+PTX(10nM)、甲硫氨酸(75μg/mL)+PTX(10nM)、甲硫氨酸(150μg/mL)+PTX(10nM)。37℃孵育48小时后,向各孔中加入10μL CCK-8(凯基生物)检测溶液,37℃下孵育1~4小时,摇床上温柔混匀,酶标仪在450nm波长处检测每孔的吸光度。
细胞存活率(%)=[(As-Ab)/(Ac-Ab)]×100;
As=实验孔吸光度(含有细胞,培养基,CCK-8和受试化合物的孔的吸光度);
Ab=空白孔吸光度(含有培养基和CCK-8的孔的吸光度);
Ac=对照孔吸光度(含有细胞,培养基和CCK-8的孔的吸光度)。
测定结果表明,在MCF-7荷瘤小鼠模型中,甲硫氨酸联合紫杉醇给药组小鼠的肿瘤体积有非常显著的变小趋势;CCK-8结果显示,补充甲硫氨酸显著增强紫杉醇对肿瘤细胞的抑制作用。由此可见,紫杉醇合并甲硫氨酸补充能够显著抑制肿瘤细胞的增长,即甲硫氨酸可显著增强紫杉醇的抗肿瘤药效。
Claims (6)
1.甲硫氨酸在制备用于增强抗肿瘤药治疗效果新药物的应用。
2.根据权利要求1所述甲硫氨酸在制备用于增强抗肿瘤药治疗效果新药物的应用,其特征在于:所述抗肿瘤增效药物为紫杉醇。
3.根据权利要求2所述甲硫氨酸在制备用于增强抗肿瘤治疗效果新药物的应用,其特征在于:所述抗肿瘤增效药物除了紫杉醇外,还可以是其它抗肿瘤药物,如多西他赛、阿霉素、表阿霉素等抗肿瘤药物。
4.根据权利要求3所述甲硫氨酸在制备用于增强抗肿瘤治疗效果新药物的应用,其特征在于:所述肿瘤为乳腺癌。
5.根据权利要求4所述甲硫氨酸在制备用于增强抗肿瘤治疗乳腺癌效果新药物的应用,其特征在于:所述肿瘤除乳腺癌外,还可以是非小细胞肺癌、肝癌、结肠癌、胃癌、急性或慢性粒性白血病等肿瘤。
6.根据权利要求5所述甲硫氨酸在制备用于增强抗肿瘤治疗效果新药物的应用,其特征在于:甲硫氨酸可以作为单独成分与抗肿瘤药物分开使用,或将甲硫氨酸与抗肿瘤药物合并使用,或2者以一定配比制备成为复方药物使用。
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CN113577058A (zh) * | 2021-09-03 | 2021-11-02 | 丽水市中心医院 | 吡喃酮化合物在制备肺癌化疗增敏药物中的应用及药物 |
CN113577058B (zh) * | 2021-09-03 | 2023-03-14 | 丽水市中心医院 | 吡喃酮化合物在制备肺癌化疗增敏药物中的应用及药物 |
CN115227688A (zh) * | 2022-07-06 | 2022-10-25 | 上海纳米技术及应用国家工程研究中心有限公司 | 双硫仑在增敏紫杉醇对肺癌治疗效果中的应用 |
CN118267400A (zh) * | 2024-05-31 | 2024-07-02 | 山东德升生物工程有限公司 | 一种运载紫杉醇的自然杀伤细胞外囊泡的制备方法与应用 |
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