CN115806623A - 一种Cathepsin L抑制组合物在细胞扩增中的用途 - Google Patents
一种Cathepsin L抑制组合物在细胞扩增中的用途 Download PDFInfo
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Abstract
本发明涉及一种CathepsinL抑制组合物在细胞扩增中的用途。本发明制备获得了重组CathepsinL蛋白,将所述蛋白免疫小鼠制备获得了相应的单克隆抗体,所述抗体具有较好的结合和抑制蛋白活性的能力同时能够抑制肝癌的活力,在制药领域具有广阔的应用前景。
Description
技术领域
本申请涉及生物领域,更具体的涉及一种Cathepsin L抑制组合物在细胞扩增中的用途。
背景技术
组织蛋白酶(cathepsin)在1920年由Lanner首次发现,它广泛存在于动物各种组织细胞内,在维持动物正常生命活动中扮演着重要角色。根据酶的催化机制,组织蛋白酶分为以下4大类:金属蛋白酶、丝氨酸蛋白酶、天冬氨酸蛋白酶和半胱氨酸蛋白酶。除此之外,还有谷氨酸蛋白酶、苏氨酸蛋白酶和一些尚未进行分类的蛋白酶。根据组织蛋白酶代表底物还可以分为L、B1、D、E、F、FG、GF、G、H、S等几大类。
Cathepsin L是半胱氨酸蛋白酶家族的重要成员之一,广泛分布于人体各种组织细胞的溶酶体中。CathepsinL的蛋白分子量为30KD,当溶酶体膜不稳定时可释放至胞浆,被溶酶体中其他的蛋白水解酶激活或自身激活,形成24KD的活性形式。Cathepsin L主要功能是降解各种组织蛋白,水解某些前体蛋白(酶原和激素原),生成其活性形式,或激活其他蛋白水解酶系统,从而参与机体的多种生理活动,如抗原递呈、激素前体的活化、骨基质的降解等。中枢神经系统中,Cathepsin L主要由激活的小胶质细胞产生和释放。
溶酶体CTSL是多种肿瘤疾病中的标志物和潜在治疗靶标的蛋白水解酶。根据研究表明,癌细胞通过分泌蛋白酶降解细胞外膜成分,从而促进肿瘤的侵袭和转移。对3种鼠黑素瘤变体的观察发现,癌细胞通过CTSL mRNA的持久翻译来维持CTSL的高表达水平,而肿瘤细胞本身就是CTSL mRNA表达升高的来源。测定肿瘤相邻的非恶性组织和远离肿瘤的非恶性组织的匀浆中几种蛋白酶样肽酶的活性,发现肿瘤相邻的A-NM中CTSL的活性显着高于D-NM组织。与正常组织相比,在胃癌、结直肠癌、乳腺癌和甲状腺癌中,也有CTSL酶活性增加的报道,这提示CTSL表达上调与疾病进展有密切关系。同时研究发现CTSL参与卵巢癌细胞的增殖和侵袭,CTSL在卵巢癌(ovariancancer,OC)中过度表达,CTSL的下调则能显著抑制了人卵巢癌细胞(skov3)的增殖和侵袭能力,而CTSL在ov90细胞中的上调则会产生相反的效果。与OC细胞相比,在裸鼠体内CTSL沉默细胞发育成肿瘤的能力降低,而这些细胞的异种移植瘤生长明显受到抑制。大量文献资料表明癌症组织CTSL的表达水平显著高于正常组织,其中肾和睾丸肿瘤表达的CTSL水平最高,多数乳腺癌表达的CTSL水平较高。CTSL的过表达与人类动脉粥硬化和主动脉瘤(aorticaneurysm,AA)的发生发展同样有着密切联系。重组半胱氨酸蛋白酶抑制剂C具有较高的抗蛋白酶活性,主要通过抑制CTSL活性,达到抑制癌细胞的生长和侵袭的效果。在主动脉瘤中,当不存在半胱氨酸蛋白酶抑制剂C时,CTSL的活性增强,促进了微血管形成、细胞凋亡、白细胞黏附及细胞增殖,从而导致AA病变面积的增大以及动脉管腔直径增大。因此CTSL和半胱氨酸蛋白酶抑制剂C可能与AA存在相关关系。以CTSL结构特点为治疗肿瘤的突破点,中国科学家发现阿斯非芬酯及其类似物能与CTSL形成氢键,与其紧密结合,从而抑制癌细胞的增殖和迁徙。总之,通过敲除卵巢癌细胞和乳腺癌细胞的CTSL基因来减少肿瘤细胞的增殖、迁移和侵袭,增强胶质瘤细胞对放射治疗敏感性。
目前,针对Cathepsin L抑制剂主要包括Clik148、Z-FY-DMK等几种类型,但是针对Cathepsin L生物类的抑制剂特别是Cathepsin L单克隆抗体的研究还不够多。
发明内容
本发明针对现有技术的缺陷,提供一种特异性的Cathepsin L蛋白单克隆抗体。
所述单克隆抗体是通过免疫小鼠制备获得的。
具体的,所述单克隆抗体亚型是IgG1类型的。
更进一步的,所述单克隆抗体具有亲和常数K值大于1×109mol/L。
更进一步的,所述单克隆抗体具有亲和常数K值大于2×109mol/L。
更进一步的,所述单克隆抗体具有亲和常数K值大于3×109mol/L。
更进一步的,所述单克隆抗体具有亲和常数K值大于4×109mol/L。
更进一步的,所述单克隆抗体具有亲和常数K值大于5×109mol/L。
更进一步的,所述单克隆抗体具有亲和常数K值大于6×109mol/L。
更进一步的,所述单克隆抗体的抗体重链可变区序列和抗体轻链可变区序列分别如SEQ ID NO:1和SEQ ID NO:2所示。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在80%的同源性,仍保持抗体活性。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在85%的同源性,仍保持抗体活性。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在90%的同源性,仍保持抗体活性。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在95%的同源性,仍保持抗体活性。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在96%的同源性,仍保持抗体活性。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在97%的同源性,仍保持抗体活性。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在98%的同源性,仍保持抗体活性。
更进一步的,所述单克隆抗体是在抗体重链可变区序列和抗体轻链可变区序列分别在SEQ ID NO:1和SEQ ID NO:2的基础上,可以进行1或2个或3个或4个或5个或6个或多个的氨基酸取代或突变,并且保持在99%的同源性,仍保持抗体活性。
进一步的,本发明还提供了Cathepsin L蛋白单克隆抗体在制备用于抑制癌细胞增殖的药物组合物中的用途。
进一步的,所述癌症是常见的癌症类型包括:胆管癌、膀胱癌、骨癌、肠癌(包括结肠癌和直肠癌)、脑癌、乳腺癌、神经内分泌系统癌(通常称为类癌)、宫颈癌、眼癌、食道癌、头颈部癌(该组包括起始于形成口腔、鼻、喉、耳的内层(lining)或覆盖舌头的表层的细胞的癌)、卡波西肉瘤、肾癌、喉癌、白血病、肝癌、肺癌、淋巴结癌、霍奇金淋巴瘤、非霍奇金淋巴瘤、黑素瘤、间皮瘤、骨髓瘤、卵巢癌、胰腺癌、阴茎癌、前列腺癌、皮肤癌、软组织肉瘤、脊髓癌、胃癌、睾丸癌、甲状腺癌、阴道癌、外阴癌和子宫癌。
进一步的,所述肝癌细胞是SMMC-7721肝癌细胞。
在一些实施方案中,向患有癌症的受试者施用本发明的组合物抑制肿瘤生长,从而治疗受试者。在一些实施方案中,抑制肿瘤生长是指在施用本发明的组合物后,减缓或防止肿瘤大小的增长。在一些实施方案中,将肿瘤生长与本领域已知的治疗癌症的相关临床数据进行比较。在一些实施方案中,将肿瘤生长与治疗的受试者的治疗前的肿瘤大小和/或体积进行比较。通过本领域已知的方法测量肿瘤的大小。
进一步的,本发明的药物组合物还包含药学上可接受的载体。
进一步的,本发明的药物组合物还包含第二治疗剂。
有益效果
本发明制备获得了重组Cathepsin L蛋白,将所述蛋白免疫小鼠制备获得了相应的单克隆抗体,所述抗体具有较好的结合和抑制蛋白活性的能力同时能够抑制肝癌的活力,在制药领域具有广阔的应用前景。
附图说明
图1Cathepsin L重组蛋白电泳图
图2Western-blot检测C7I6单克隆抗体特异性结果图,泳道1是小鼠皮肤细胞,泳道2为SMMC-7721肝癌细胞。
图3单抗处理后Cathepsin L/GAPDH比值结果图
具体实施方式
下面将参照附图更详细地描述本发明的具体实施例。虽然附图中显示了本发明的具体实施例,然而应当理解,可以以各种形式实现本发明而不应被这里阐述的实施例所限制。相反,提供这些实施例是为了能够更透彻地理解本发明,并且能够将本发明的范围完整的传达给本领域的技术人员。
实施例1Cathepsin L重组蛋白的制备
根据人CathepsinL基因,设计引物由由上海生物工程有限公司合成。引物序列:
上游引物:5′-GCCTCGAGCATGAATCCTACACTCATCCTTG-3′,引入了一个XhoI酶切位点;
下游引物:5′-GCAGGATCCTCACACAGTGGGGTAGC-3′,引入了一个BamHI酶切位点。
以人卵巢癌细胞skov3为模板,提取总RNA,逆转录cDNA,然后进行PCR扩增。PCR总体积50μL,内含cDNA模板1μL,引物(10μmol/L)各2μL,dNTP(10mmol/L)4μL,EX-Taq酶(5U/μL)0.25μL,10×PCRbuffer 5μL,去离子水35.75μL。反应条件:94℃预变性5min;94℃变性60s,55℃退火45s,72℃延伸60s,共30个循环;72℃延伸10min。取PCR产物纯化后连接PMD18-T载体,pcDNA3.1载体和测序正确的pMD18-T-CTSL质粒分别用BamH I、Xho I进行双酶切反应,然后在T4连接酶的作用下进行连接,获得重组质粒,命名为pcDNA3.1-CTSL。将质粒转化CHO细胞,选择阳性细胞,进行扩大培养,接种至旋转瓶,至细胞密度达到90%以上时更换为无血清培养基,收获上清,通过亲和层析rProteinA柱纯化,对纯化收获的蛋白进行SDS-PAGE鉴定。结果如图1所示,制备并获得了较为纯净的Cathepsin L重组蛋白,调整蛋白浓度为5mg/mL。
实施例2Cathepsin L单克隆抗体的研制
背部皮下注射实施例1制备的Cathepsin L重组蛋白给7周龄的雌性BALB/c小鼠。第1次免疫,免疫剂量为100μg/只,用生理盐水稀释到适当体积,再加等体积弗式完全佐剂(FCA),采用注射器推拉法乳化成油包水(W/O)状态,将乳化好的抗原进行背部皮下多点注射(100μL/只);2周后,以相同剂量的免疫原与弗式不完全佐剂(FIA)混合乳化,进行加强免疫,此后,应用同样方法进行第3、4次免疫,每次间隔14d。每次免疫后1周,小鼠尾静脉采血分离血清,检测血清效价,以待测孔OD值与阴性对照孔OD值(N)的比值P/N≥2.1的最小值的血清稀释度为待测血清效价。效价达到1:10000后取效价最高小鼠的脾细胞与小鼠骨髓瘤细胞株SP2/0进行融合。融合前3d不加佐剂,腹腔直接注射抗原100μg冲击免疫。融合前先将1mL的PEG-1500和20mL的RPMI-1640基础培养基于37℃预温。取效价最高小鼠的脾细胞与骨髓瘤细胞按10:1比例混合于50mL离心管内,1000r/min离心10min。离心后弃上清。然后轻轻敲击离心管底使细胞松散。在37℃水浴中沿管壁加入PEG 1mL,50s内加完,边加边转动离心管,不宜吹打,加完后在37℃水浴中静置反应1min,再加入RPMI-1640基础培养基以终止PEG的作用,在1min内加完1mL,再在1min内加完2mL,然后逐渐加快速度,在5min内加完15mL。加终止液时应缓慢加入,边加边轻轻搅拌。不宜吹打以免使刚融合的细胞分离。融合细胞终止后于800r/min离心5min,并吸干以防残留PEG。再加入预温的HAT培养基至所需的细胞浓度,并轻轻将细胞混匀,然后100μL/孔滴板。细胞融合7d后,用间接ELISA法检测抗体,获得抗体分泌阳性孔35孔,用HT培养基扩大培养后,采用有限稀释法接种于96孔培养板内,选择单个细胞集落孔的培养上清作抗体检测,阳性者用同样方法再次克隆,直至克隆后有杂交瘤细胞生长的抗体分泌阳性率为100%,其中阳性反应最强的杂交瘤细胞命名为C7I6。
采取体内诱生法制备大量腹水。取10周龄BALB/c小鼠腹腔注射弗氏不完全佐剂每只0.2mL,1周后每只小鼠腹腔注射1×106个C7I6杂交瘤细胞,待注射后10d左右小鼠腹部明显隆起时收集腹水,4000r/min离心10min,吸取上清分装,采用辛酸-饱和硫酸铵沉淀法纯化抗体备用。
实施例3Western-blot检测C7I6单克隆抗体的特异性
用蛋白裂解液裂解SMMC-7721肝癌细胞和小鼠皮肤细胞,提取蛋白质,Bradford法定量总蛋白浓度,10%SDS-PAGE分离蛋白样品(上样量为50μg),半干法转移至醋酸纤维素膜,常温下5%BSA封闭2h,Cathepsin L一抗C7I6(1:1000)孵育2h,HRP标记二抗(1∶5000)作用1h,加入化学发光试剂后置于胶片下曝光,显影定影后分析,以β-actin作为内对照。结果如图2所示。
从图2可以看出,本发明制备的Cathepsin L单克隆抗体能够与人细胞中的Cathepsin L蛋白特异性结合,而不与小鼠细胞内的其他蛋白或者小鼠体内的Cathepsin L蛋白结合,说明本发明的单克隆抗体具有较好的特异性。
实施例4C7I6单克隆抗体生物学特异性和序列鉴定
亚型检测:选用Mouse Immunoglobulin Isotyping ELISA试剂盒(BD),按操作说明,羊抗鼠IgG板子包被、封闭。取杂交瘤上清,进行检测。结果显示,C7I6单克隆抗体为IgG1亚型鼠源单克隆抗体。
亲和力鉴定:用CBS液将Cathepsin L蛋白稀释成浓度1μg/mL和2μg/mL的包被液分别包被酶标板,通过间接ELISA法测定单抗腹水效价,以单抗浓度为横坐标,OD450值为纵坐标,绘出相应的2条间接ELISA反应曲线,以每条曲线上部平坦段的OD450值作为100%,在曲线上算出50%OD450值时对应的抗体浓度。按照公式Kaff=(n-1)/2(n[Ab′]t-[Ab]t)计算单抗的亲和常数,其中n=[Ag]t/[Ag′]t,[Ag]t、[Ag′]t为2个不同的包被原浓度,[Ab]t、[Ab′]t为各包被原浓度下50%OD450值对应的抗体浓度。根据亲和力测定结果计算出C7I6单克隆抗体亲和常数K值为6.45×109mol/L。
取培养的杂交瘤细胞系1×106,离心取上清,将细胞沉淀委托苏州金唯智生物科技有限公司进行抗体重链和轻链可变区序列测定,得到抗体重链可变区序列和抗体轻链可变区序列分别如SEQ ID NO:1和SEQ ID NO:2所示。
实施例5C7I6单克隆抗体对癌细胞的抑制作用
将SMMC-7721肝癌细胞冻存管在37℃温水中快速摇晃融化,时间1min左右,加入4-5ml培养基混匀。在1000RPM左右条件下离心4min,弃上清,加1-2ml DMEM/F12培养基吹匀,将细胞悬液加入培养瓶中,补加适量培养基。细胞密度达到80-90%时即可传代,弃去培养上清,用PBS或生理盐水清洗1-2次;加入2ml0.25%胰酶(T25瓶),使胰酶覆盖整个瓶或皿,盖好放入培养箱消化;1-2min后,显微镜下观察细胞,若大部分细胞回缩且有少量细胞脱落,轻轻吹打下确认消化情况后加入完全培养基终止消化;若细胞还是贴壁,放回培养箱继续消化至可以轻轻吹打下为止;将细胞悬液1000RPM左右条件下离心4min,弃上清;用新鲜培养基重悬后加入培养瓶或皿中,T25培养瓶加6-8ml培养基;悬浮细胞直接离心收集,细胞沉淀重悬后分到新培养瓶中。带细胞长到对数生长期,调整细胞浓度为1×105个/ml,接种于96孔培养板中,每孔100μl,分别加入终浓度为0、10、20、50、100、200μg/mL的单克隆抗体,设置阳性对照为50μg/mL盐酸吉西他滨,处理24h。弃上清,每孔加入MTT 10mg/ml 20μl,4h后弃去液体,每孔加入二甲基亚砜150μl,震荡10min,酶标仪测490nm处吸光度值,各孔吸光度值减去空白空吸光度值得到实际吸光度值。结果如表1所示。
表1肝癌细胞MTT细胞活力检测结果
组别 | 活力结果 |
空白组 | 0.458±0.039 |
10μg/mL的单克隆抗体 | 0.287±0.027* |
20μg/mL的单克隆抗体 | 0.265±0.018* |
50μg/mL的单克隆抗体 | 0.243±0.026* |
100μg/mL的单克隆抗体 | 0.220±0.019* |
200μg/mL的单克隆抗体 | 0.196±0.016* |
阳性对照组 | 0.252±0.024* |
从表1的结果可以看出,本发明的单克隆抗体具有剂量依赖性的降低细胞活力的效果,并且与空白组相比差异极其显著(P<0.01)。与阳性对照组相比,本发明的单克隆抗体具有更好的抑制细胞活力的效果,可以在癌症治疗中具有更重要的应用。
将各组处理后的细胞,提取细胞总蛋白,进行蛋白含量检测,常规法进行WB检测,化学发光法显色,统计Cathepsin L/GAPDH比值,结果如图3所示。从图3可以看出。本发明的单克隆抗体能够较好的抑制Cathepsin L蛋白的表达,而阳性对照对该蛋白的影响较好。这说明,本发明的单克隆抗体能够特异性的抑制Cathepsin L的表达,抑制率大于88%,效果显著。
在本说明书的描述中,参考术语“一个实施例”、“一些实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本发明的至少一个实施例或示例中。在本说明书中,对上述术语的示意性表述不一定指的是相同的实施例或示例。而且,描述的具体特征、结构、材料或者特点可以在任何的一个或多个实施例或示例中以合适的方式结合。尽管已经示出和描述了本发明的实施例,本领域的普通技术人员可以理解:在不脱离本发明的原理和宗旨的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由权利要求及其等同物限定。
Claims (5)
1.一种特异性的CathepsinL蛋白单克隆抗体,其特征在于所述所述单克隆抗体重链可变区序列如SEQ ID NO:1所示,轻链可变区序列如SEQ ID NO:2所示,所述抗体具有抑制CathepsinL蛋白的活性。
2.如权利要求1所述的CathepsinL蛋白单克隆抗体在制备用于抑制CathepsinL蛋白高表达的试剂中的用途,所述试剂用于离体的细胞实验。
3.如权利要求1所述的CathepsinL蛋白单克隆抗体在制备用于抑制肝癌细胞增殖的组合物中的用途,其中肝癌是SMMC-7721肝癌细胞。
4.如权利要求3所述的用途,其特征在于所述组合物中还添加有缓冲液和防腐剂。
5.如权利要求3所述的用途,其特征在于组合物是液体制剂形式。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101735317A (zh) * | 2008-11-25 | 2010-06-16 | 中国科学院北京基因组研究所 | Cathepsin D抗原多肽、应用及含有该多肽的检测试剂盒 |
CN102423489A (zh) * | 2011-11-30 | 2012-04-25 | 苏州大学 | 组织蛋白酶l抑制剂在制备放疗增敏药物中的应用 |
CN112552396A (zh) * | 2020-12-30 | 2021-03-26 | 河南中泽生物工程有限公司 | 抗非洲猪瘟病毒p54蛋白单克隆抗体、制备方法及应用 |
WO2022083603A1 (zh) * | 2020-10-22 | 2022-04-28 | 上海良润生物医药科技有限公司 | 半胱氨酸蛋白酶抑制剂与Cathepsin复合物作为肿瘤诊断标志物的应用 |
-
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101735317A (zh) * | 2008-11-25 | 2010-06-16 | 中国科学院北京基因组研究所 | Cathepsin D抗原多肽、应用及含有该多肽的检测试剂盒 |
CN102423489A (zh) * | 2011-11-30 | 2012-04-25 | 苏州大学 | 组织蛋白酶l抑制剂在制备放疗增敏药物中的应用 |
WO2022083603A1 (zh) * | 2020-10-22 | 2022-04-28 | 上海良润生物医药科技有限公司 | 半胱氨酸蛋白酶抑制剂与Cathepsin复合物作为肿瘤诊断标志物的应用 |
CN112552396A (zh) * | 2020-12-30 | 2021-03-26 | 河南中泽生物工程有限公司 | 抗非洲猪瘟病毒p54蛋白单克隆抗体、制备方法及应用 |
Non-Patent Citations (2)
Title |
---|
JOHN P DALTON 等: "Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines", INTERNATIONAL JOURNAL FOR PARASITOLOGY, vol. 33, no. 11, pages 1173 - 1181 * |
晋云;董家鸿;: "肝细胞癌cathepsin L表达分析", 第三军医大学学报, no. 22, pages 2139 - 2141 * |
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