CN111110858A - 一种Her2靶向的多肽药物偶联物及其制备方法和应用 - Google Patents
一种Her2靶向的多肽药物偶联物及其制备方法和应用 Download PDFInfo
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- CN111110858A CN111110858A CN201811282295.0A CN201811282295A CN111110858A CN 111110858 A CN111110858 A CN 111110858A CN 201811282295 A CN201811282295 A CN 201811282295A CN 111110858 A CN111110858 A CN 111110858A
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Abstract
Description
技术领域
本发明涉及一种Her2靶向的多肽药物偶联物及其制备方法和应用,尤其涉及一种可应用于肿瘤治疗的Her2靶向的多肽药物偶联物及其制备方法和应用。
背景技术
恶性肿瘤是一种由多种诱因共同作用而引起的严重威胁人类健康的常见病和多发病,占所有疾病死亡率的第二位,严重危害着我国人民的健康,给患者家庭和国家医疗体系带来严重的经济负担,也对新型抗肿瘤药物的研发提出了严峻的挑战,但取得的成效却不尽如人意,主要原因包括肿瘤易产生耐药性,化疗药物对肿瘤的靶向性差,毒副作用大等。
小分子靶向肽可以通过受体相互作用将药物选择性地靶向到肿瘤部位,使治疗药物在病灶区浓集,病灶区的药物浓度超出常规制剂的数倍乃至数百倍,提高了系统化疗的效率,大大地降低了药物的毒副作用,例如LTVSPWY 多肽能够与肿瘤细胞表面Her2受体特异性结合。传统的化疗药物水溶性差,在多肽链的末端引入强亲水性的精氨酸或者赖氨酸能够改善化疗药物的水溶性。
细胞内的谷胱甘肽浓度(0.5~10mM/L)是细胞外谷胱甘肽浓度(2~ 20μM/L)的200倍以上,二硫键在一定量的谷胱甘肽(GSH)或二硫苏糖醇(DTT)等还原剂存在下被还原生成巯基,但是在人体的正常体温、pH 和氧化等环境下非常稳定,即细胞外的谷胱甘肽浓度不足以还原二硫键,另外,肿瘤组织细胞比正常组织细胞缺氧,更具有还原性环境。因此,可以将亲水性的多肽链和疏水性的药物通过二硫键链接,药物进入靶细胞后被GSH还原,即二硫键断裂生成巯基,从而快速有效地释放药物,并扩散到细胞核等结构,从而杀死癌细胞。目前最常用的二硫键连接臂主要为2,2′- 二硫代二乙酸、3,3′-二硫代二丙酸等,例如以3,3′-二硫代二丙酸作为连接臂的药物前体,在GSH、DTT等还原条件下,二硫键迅速发生断裂,但是释放的药物通常会带着“尾巴”,而非原药分子形式。
发明内容
本发明针对现有抗肿瘤药物选择性差、水溶性差以及还原响应释放原药并非原药分子形式等方面存在的不足,提供一种Her2靶向的多肽药物偶联物及其制备方法和应用。
本发明解决上述技术问题的技术方案如下:
一种Her2靶向的多肽药物偶联物,具有如式Ⅰ所示的分子结构:
其中,Aaa1为L或D型的Lys或Arg;Aaa2L或D型的Lys或Arg;X 为CH2、NH或O;ROH为疏水性抗肿瘤药物;n为1或2。
进一步,所述Her2靶向的多肽药物偶联物优选Aaa1为L型Arg,Aaa2为L型Arg,具有如式Ⅱ所示的结构式:
其中,X为CH2、NH或O;ROH为疏水性抗肿瘤药物;n为1或2。
进一步,所述抗肿瘤药物是指紫杉烷类、喜树碱类、长春碱类药物中的任意一种。
进一步,所述紫杉烷类药物是指紫杉醇、多烯紫杉醇、卡巴他赛、拉洛他赛中的一种,所述喜树碱类药物是指喜树碱、伊立替康、拓扑替康、10- 羟基喜树碱、7-乙基-10-羟基喜树碱中的一种,所述长春碱类药物是指长春碱、长春新碱、长春瑞滨中的一种。
本发明提供的多肽药物偶联物释放原药分子的机理为:
本发明采用了肿瘤微环境靶向给药的思路,通过引入对还原环境敏感的自毁式二硫键连接臂,在肿瘤细胞内谷胱甘肽提供的还原条件下,多肽药物偶联物的二硫键发生断裂,二硫键断裂后生成一个自由的巯基,由于受到结构稳定因素的影响,该巯基会亲核进攻邻近的酯键从而生成一个稳定的五元环或六元环结构,同时释放抗癌药物的原药分子,以卡巴他赛为例,其还原响应释放原药分子的机理如图4所示。
本发明提供的靶向多肽药物偶联物的有益效果为:
1)能够实现靶向给药,靶向多肽可将抗肿瘤药物运送到特定的肿瘤细胞,提高了药物的靶向性,增加了药物疗效,降低对正常细胞的毒副作用;
2)本发明提供的多肽药物偶联物发挥了二硫键在肿瘤部位特异性降解的特点,与常规的2,2′-二硫代二乙酸、3,3′-二硫代二丙酸等连接臂相比,无需通过进一步水解就可以得到原药分子形式的抗癌药物。
本发明还要求保护前述的靶向多肽药物偶联物在抗肿瘤药物领域的应用。
进一步,所述肿瘤指的是胃癌和乳腺癌。
本发明所述Her2靶向的多肽药物偶联物的制备方法如方法一或方法二所示:
方法一包括如下步骤:
1)以含有2~3个碳原子的吡啶基二硫基醇或吡啶基二硫基胺为原料,与对硝基氯甲酸苯酯进行偶联反应制备碳酸酯或氨基碳酸酯;
2)将步骤1)所得碳酸酯或氨基碳酸酯与含羟基基团的疏水性抗肿瘤药物通过酯交换反应生成偶联物;
3)将步骤2)所得的偶联物与靶向多肽通过巯基-二硫键交换反应,制备含有二硫键的多肽药物偶联物;
反应路线如下:
其中,X为NH或O;ROH为疏水性抗肿瘤药物;n为1或2;
方法二包括如下步骤:
1)以4-(2-吡啶基二硫基)丁酸或5-(2-吡啶基二硫基)戊酸为原料,与含有羟基的疏水性抗肿瘤药物发生酯化反应,生成疏水性抗肿瘤药物的酯化产物;
2)将步骤1)所得的酯化产物与靶向多肽通过巯基-二硫键交换反应,制备含有二硫键的多肽药物偶联物;
反应路线如下:
其中,ROH为疏水性抗肿瘤药物;n为1或2。
进一步,方法一的具体操作步骤如下:
步骤1):在碱性条件下,将吡啶基二硫基醇或吡啶基二硫基胺溶于溶剂中得溶液,室温搅拌下将对硝基氯甲酸苯酯滴加至溶液中,控制吡啶基二硫基醇或吡啶基二硫基胺与对硝基氯甲酸苯酯的摩尔比为1:(1~3),室温反应1~ 10小时,反应液经后处理得碳酸酯或氨基碳酸酯;
步骤2):在碱性条件下,将步骤1)所得的碳酸酯或氨基碳酸酯和含有羟基基团的疏水性抗肿瘤药物按摩尔比1:(1~3)溶于溶剂中,回流反应1~10 小时,降至室温,反应液经后处理得二者的偶联物;
步骤3):惰性氛围中,搅拌条件下将靶向多肽滴加到步骤2)所得的偶联物的溶液中,靶向多肽和偶联物的摩尔比为1:(1~3),室温下反应12~ 48小时,经后处理得含有二硫键的靶向多肽药物偶联物。
进一步,方法二的具体操作步骤如下:
步骤1):以4-(2-吡啶基二硫基)丁酸或5-(2-吡啶基二硫基)戊酸为原料,与含有羟基基团的疏水性药物按摩尔比(1~3):1溶于溶剂中,室温下反应 12~36小时,反应液经水洗、分液、真空干燥得疏水性抗肿瘤药物的酯化产物;
步骤2):惰性氛围中,搅拌条件下将靶向多肽滴加到步骤1)所得的酯化产物的溶液中,靶向多肽和酯化产物的摩尔比为1:(1~3),室温下反应12~ 48小时,经后处理得含有二硫键的靶向多肽药物偶联物。
附图说明
图1为实施例1所得的Her2靶向的多肽卡巴他赛偶联物的 MALDI-TOF-MS图;
图2为实施例1所得Her2靶向的多肽卡巴他赛偶联物与卡巴他赛对胃癌细胞N87的增殖抑制活性比较;
图3为实施例1所得Her2靶向的多肽卡巴他赛偶联物与卡巴他赛对乳腺癌细胞BT474的增殖抑制活性比较。
图4为多肽药物偶联物还原响应释放原药分子的机理示意图。
具体实施方式
以下结合实例对本发明的原理和特征进行描述,所举实例只用于解释本发明,并非用于限定本发明的范围。
实施例1:
一种Her2靶向的多肽卡巴他赛偶联物,具有如下结构式:
使用Fmoc策略的固相多肽合成方法,使用CSBio公司生产的多肽合成仪进行本发明的靶向多肽的合成。
LTVSPWYCRR合成所用试剂选择:
(1)载体树脂:Fmoc-Arg(Pbf)Wang,取代度:0.67
(2)所选用的保护氨基酸:Fmoc-Leu-OH,Fmoc-Thr(tBu)-OH, Fmoc-Val-OH,Fmoc-Ser(tBu)-OH,Fmoc-Pro-OH,Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH,Fmoc-D-Cys(Trt)-OH,Fmoc-Arg(pbf)-OH,反应中所用保护氨基酸3倍过量。
(3)本发明所使用的脱保护试剂为:哌啶/N,N-二甲基甲酰胺,比例为 20:80。
(4)本发明所使用的偶联试剂为:DIEA/HBTU。
(5)本发明中所使用的切割试剂为:TFA/三异丙基硅烷/水/1,2-乙二硫醇,比例为:94:2:2:2。用10倍体积乙醚沉淀,将制得的多肽粗品以C18制备柱进行纯化,检测波长:214nm,流动相A:乙腈(含0.1%三氟乙酸),流动相B:水(含0.1%三氟乙酸)。MS-MALDI-TOF确定所得纯品分子量,HPLC 确定样品纯度,经冷冻干燥得到目标多肽。
上述Her2靶向的多肽卡巴他赛偶联物的制备方法如下:
1)4-(2-吡啶基二硫基)丁酸的制备:
将4-溴丁酸(10g,59.9mmol)和硫脲(6.37g,83.8mmol)加入250mL 乙醇中,回流4小时。将NaOH(24g,600mmol)溶于250mL乙醇后,加入到反应体系中,继续回流16小时。冷却至室温,过滤得到白色固体,加水溶解。水相用乙醚洗,分液,以2mol/L的HCl调至pH=1后,乙醚萃取,无水硫酸钠干燥,真空干燥得4.9g淡黄色油状物4-巯基丁酸,收率为 68%。将4-巯基丁酸(4.9g,40.8mmol)和二硫二吡啶(Py-SS-Py,18g, 81.7mmol)溶于60mL甲醇中,室温下反应3小时。减压蒸去甲醇,过中性氧化铝柱分离,浓缩,真空干燥得5.4g淡黄色油状物,收率58%。
2)4-(2-吡啶基二硫基)丁酸与卡巴他赛偶联物的制备:
将卡巴他赛(1.0g,1.2mmol),4-(2-吡啶基二硫基)丁酸(0.3g,1.3 mmol),4-二甲氨基吡啶(DMAP,0.18g,1.5mmol),N,N-二异丙基乙胺 (DIEA,0.25ml,1.5mmol)溶于50mLDMF中,冰浴下加入1-乙基-3- (二甲基氨基丙基)碳酰二亚胺盐酸盐(EDCI,0.3g,1.5mmol),室温下反应48小时。反应液中加入二氯甲烷(DCM),依次以稀盐酸、饱和碳酸氢钠、饱和食盐水洗有机相,分液,干燥后,减压蒸去溶剂。经硅胶柱纯化,浓缩,真空干燥得0.95g黄色固体,收率为75%。
3)LTVSPWYCRR-卡巴他赛的制备:
将LTVSPWYCRR多肽(40mg,0.031mmol)溶于2mL DMSO中,氮气保护、搅拌下,将Py-SS-Caba(49mg,0.047mmol)的DMSO(2mL) 溶液加入其中,室温下反应24小时。将反应液直接进行制备液相纯化:检测波长:254nm,流动相A:乙腈(含0.1%三氟乙酸),流动相B:水(含0.1%三氟乙酸)。MS-MALDI-TOF测试,表明得到LTVSPWYCRR-卡巴他赛偶联物(多肽-二硫键-卡巴他赛),经冷冻干燥得到目标偶联物20mg,收率30%。
合成路线如下所示:
实施例2:
一种Her2靶向的多肽SN-38偶联物,具有如下结构式:
LTVSPWYCKR合成所用试剂选择:
(1)载体树脂:Fmoc-Arg(Pbf)Wang,取代度:0.67
(2)所选用的保护氨基酸:Fmoc-Leu-OH,Fmoc-Thr(tBu)-OH, Fmoc-Val-OH,Fmoc-Ser(tBu)-OH,Fmoc-Pro-OH,Fmoc-Trp(Boc)-OH,Fmoc-Tyr(tBu)-OH,Fmoc-D-Cys(Trt)-OH,Fmoc-Lys(Boc)-OH,Fmoc-Arg (pbf)-OH,反应中所用保护氨基酸3倍过量。
(3)本发明所使用的脱保护试剂,偶联试剂,切割试剂均与实施例1相同。制得的多肽粗品以C18制备柱进行纯化,检测波长:214nm,流动相A:乙腈(含0.1%三氟乙酸),流动相B:水(含0.1%三氟乙酸)。MS-MALDI-TOF 确定所得纯品分子量,HPLC确定样品纯度,经冷冻干燥得到目标多肽。
上述多肽SN-38偶联物的制备方法如下:
1)4-硝基苯基-(2-吡啶基二硫基)乙氨基碳酸酯的制备:
将吡啶基二硫乙胺盐酸盐(1.0g,4.5mmol)和三乙胺(1.0g,9.9mmol) 溶于50mL二氯甲烷中,冰浴下将对硝基氯甲酸苯酯(1.22g,6.3mmol) 缓慢滴加至溶液中,滴毕,室温反应5小时。浓缩,硅胶柱分离、浓缩、真空干燥得淡黄色油状液体1.18g,收率75%。
2)TBDPS-7-乙基-10-羟基喜树碱的制备:
将7-乙基-10-羟基喜树碱(2.0g,5.1mmol)加入100mL二氯甲烷中,向其中加入三乙胺(3.2mL,23mmol)和TBDPSCl(5.3mL,20.4mmol),回流过夜。冷却至室温,依次以稀盐酸、饱和碳酸氢钠、饱和食盐水洗有机相,分液,干燥后,减压浓缩。将浓缩液滴入大量正己烷中进行沉淀,过滤,真空干燥得3.0g黄色固体,收率为93%。
3)TBDPS-PySS-7-乙基-10-羟基喜树碱的制备:
将TBDPS-7-乙基-10-羟基喜树碱(1.9g,3.0mmol),4-硝基苯基-(2- 吡啶基二硫基)乙氨基碳酸酯(1.18g,3.36mmol),4-二甲氨基吡啶(DMAP, 0.4g,3.36mmol),溶于200mLDCM中,室温下反应24小时。反应液依次以稀盐酸、饱和碳酸氢钠、饱和食盐水洗,分液,干燥后,减压蒸去溶剂,真空干燥得2.0g黄色固体TBDPS-PySS-7-乙基-10-羟基喜树碱,收率为80%。
4)PySS-7-乙基-10-羟基喜树碱的制备:
将TBDPS-PySS-7-乙基-10-羟基喜树碱(2.0g,2.4mmol)和TBAF(2.5 g,9.6mmol)溶于THF和0.05M HCl(体积比1:1)的混合液300mL中,室温反应。反应液以二氯甲烷萃取后,以饱和食盐水洗,无水硫酸钠干燥后,减压蒸去溶剂。经硅胶柱纯化,浓缩,真空干燥得1.2g黄色固体PySS-7- 乙基-10-羟基喜树碱,收率为84%。
5)LTVSPWYCKR-SN38的制备:
将LTVSPWYCKR多肽(42mg,0.034mmol)溶于4mL DMSO中,氮气保护、搅拌下,将PySS-7-乙基-10-羟基喜树碱(31mg,0.051mmol)的 DMSO(2mL)溶液加入其中,室温下反应24小时。将反应液直接进行制备液相纯化:检测波长:254nm,流动相A:乙腈(含0.1%三氟乙酸),流动相B:水(含0.1%三氟乙酸)。经冷冻干燥得到目标偶联物18mg,收率30%。
合成路线如下:
实施例3:
一种Her2靶向的多肽长春瑞滨偶联物,具有如下结构式:
LTVSPWYCKK合成所用试剂选择:
(1)载体树脂:Fmoc-Arg(Pbf)Wang,取代度:0.67
(2)所选用的保护氨基酸:Fmoc-Leu-OH,Fmoc-Thr(tBu)-OH, Fmoc-Val-OH,Fmoc-Ser(tBu)-OH,Fmoc-Pro-OH,Fmoc-Trp(Boc)-OH,Fmoc-Tyr(tBu)-OH,Fmoc-D-Cys(Trt)-OH,Fmoc-Lys(Boc)-OH,反应中所用保护氨基酸3倍过量。
(3)本发明所使用的脱保护试剂,偶联试剂,切割试剂均与实施例1相同。制得的多肽粗品以C18制备柱进行纯化,检测波长:214nm,流动相A:乙腈(含0.1%三氟乙酸),流动相B:水(含0.1%三氟乙酸)。MS-MALDI-TOF 确定所得纯品分子量,HPLC确定样品纯度,经冷冻干燥得到目标多肽。
上述多肽药物偶联物前体的制备方法如下:
1)4-硝基苯基-(2-吡啶基二硫基)丙醇碳酸酯的制备:
将吡啶基二硫丙醇(1.0g,5.0mmol)和三乙胺(0.6g,6mmol)溶于 40mL二氯甲烷中,冰浴下将对硝基氯甲酸苯酯(1.4g,7.0mmol)缓慢滴加至溶液中,滴毕,室温反应5小时。浓缩,硅胶柱分离、浓缩、真空干燥得淡黄色油状液体1.28g,收率70%。
2)3-(2-吡啶基二硫基)丙醇长春瑞滨碳酸酯的制备:
将4-硝基苯基-(2-吡啶基二硫基)丙醇碳酸酯(1.0g,2.7mmol)和长春瑞滨(2.1g,2.7mmol)溶于150mL二氯甲烷中,加入1ml三乙胺,然后升温至回流反应8小时。反应完毕,降至室温,用稀盐酸水洗有机相、分液、干燥、浓缩、经硅胶柱纯化、浓缩,真空干燥得1.75g淡黄色固体,收率为65%。
3)LTVSPWYCKK-长春瑞滨的制备:
合成路线如下所示:
将LTVSPWYCKK多肽(42mg,0.034mmol)溶于2mL DMSO中,氮气保护、搅拌下,将3-(2-吡啶基二硫基)丙醇长春瑞滨碳酸酯(51mg,0.05 mmol)的DMSO(2mL)溶液加入其中,室温下反应24小时。将反应液直接进行制备液相纯化:检测波长:254nm,流动相A:乙腈(含0.1%三氟乙酸),流动相B:水(含0.1%三氟乙酸)。MS-MALDI-TOF确定所得纯品分子量,表明得到LTVSPWYCKK-长春瑞滨偶联物,经冷冻干燥得到目标偶联物29mg,收率40%。
为了验证本发明所得多肽药物偶联物对肿瘤细胞的增殖抑制效果,我们取实施例1所得的多肽偶联卡巴他赛以及卡巴他赛进行了体外抗肿瘤细胞的效果对比实验,我们以胃癌细胞N87和乳腺癌细胞BT474为例分别进行了实施例1所得的多肽偶联卡巴他赛药物与卡巴他赛的增殖抑制效果实验,具体操作过程如下:
1)胃癌细胞N87
取对数生长期的细胞,调整适当的细胞密度,接种于96孔板内,100 μl/well,培养于37℃,5%CO2的培养箱内。培养过夜后给药,分别加药作用 48h。分设空白组、给药组,每组设4个复孔。体外抗胃癌效果如图2所示。从图2中可以看出,多肽-二硫键-卡巴他赛与卡巴他赛具有近似的细胞毒性, 多肽-二硫键-卡巴他赛对胃癌细胞N87的半数致死量IC50为0.260μM,有较强的抗肿瘤活性。
2)乳腺癌细胞BT474
取对数生长期的细胞,调整适当的细胞密度,接种于96孔板内,100 μl/well,培养于37℃,5%CO2的培养箱内。培养过夜后给药,分别加药作用 48h。分设空白组、给药组,每组设4个复孔。体外抗乳腺癌效果如图3所示。从图3中可以看出,多肽-二硫键-卡巴他赛与卡巴他赛具有近似的细胞毒性,多肽-二硫键-卡巴他赛对乳腺癌细胞BT474的半数致死量IC50为0.099 μM,有较强的抗肿瘤活性。
综上所述,本发明提供的多肽药物偶联物不仅水溶性好,且经过实验证明与原药具有基本相同的抗肿瘤活性,可以于48h内发挥出其对肿瘤细胞的抑制作用。
以上所述仅为本发明的较佳实施例,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (9)
3.根据权利要求1或2所述的Her2靶向的多肽药物偶联物,其特征在于,所述抗肿瘤药物是指紫杉烷类、喜树碱类、长春碱类药物中的任意一种。
4.根据权利要求3所述的Her2靶向的多肽药物偶联物,其特征在于,所述紫杉烷类药物是指紫杉醇、多烯紫杉醇、卡巴他赛、拉洛他赛中的一种,所述喜树碱类药物是指喜树碱、伊立替康、拓扑替康、10-羟基喜树碱、7-乙基-10-羟基喜树碱中的一种,所述长春碱类药物是指长春碱、长春新碱、长春瑞滨中的一种。
5.权利要求1~4中任一项所述的Her2靶向的多肽药物偶联物在抗肿瘤药物领域的应用。
6.根据权利要求5所述的应用,其特征在于,所述肿瘤是指乳腺癌和胃癌。
7.权利要求1~4中任一项所述的Her2靶向的多肽药物偶联物的制备方法,其特征在于,如方法一或方法二所示:
方法一包括如下步骤:
1)以含有2~3个碳原子的吡啶基二硫基醇或吡啶基二硫基胺为原料,与对硝基氯甲酸苯酯进行偶联反应制备碳酸酯或氨基碳酸酯;
2)将步骤1)所得碳酸酯或氨基碳酸酯与含羟基基团的疏水性抗肿瘤药物通过酯交换反应生成偶联物;
3)将步骤2)所得的偶联物与靶向多肽通过巯基-二硫键交换反应,制备含有二硫键的多肽药物偶联物;
反应路线如下:
其中,X为NH或O;ROH为疏水性抗肿瘤药物;n为1或2;
方法二包括如下步骤:
1)以4-(2-吡啶基二硫基)丁酸或5-(2-吡啶基二硫基)戊酸为原料,与含有羟基的疏水性抗肿瘤药物发生酯化反应,生成疏水性抗肿瘤药物的酯化产物;
2)将步骤1)所得的酯化产物与靶向多肽通过巯基-二硫键交换反应,制备含有二硫键的多肽药物偶联物;
反应路线如下:
其中,ROH为疏水性抗肿瘤药物;n为1或2。
8.根据权利要求7所述的制备方法,其特征在于,方法一的具体操作步骤如下:
步骤1):在碱性条件下,将吡啶基二硫基醇或吡啶基二硫基胺溶于溶剂中得溶液,室温搅拌下将对硝基氯甲酸苯酯滴加至溶液中,控制吡啶基二硫基醇或吡啶基二硫基胺与对硝基氯甲酸苯酯的摩尔比为1:(1~3),室温反应1~10小时,反应液经后处理得碳酸酯或氨基碳酸酯;
步骤2):在碱性条件下,将步骤1)所得的碳酸酯或氨基碳酸酯和含有羟基基团的疏水性抗肿瘤药物按摩尔比1:(1~3)溶于溶剂中,回流反应1~10小时,降至室温,反应液经后处理得二者的偶联物;
步骤3):惰性氛围中,搅拌条件下将靶向多肽滴加到步骤2)所得的偶联物的溶液中,靶向多肽和偶联物的摩尔比为1:(1~3),室温下反应12~48小时,经后处理得含有二硫键的靶向多肽药物偶联物。
9.根据权利要求7所述的制备方法,其特征在于,方法二的具体操作步骤如下:
步骤1):以4-(2-吡啶基二硫基)丁酸或5-(2-吡啶基二硫基)戊酸为原料,
与含有羟基基团的疏水性药物按摩尔比(1~3):1溶于溶剂中,室温下反应12~36小时,反应液经水洗、分液、真空干燥得疏水性抗肿瘤药物的酯化产物;
步骤2):惰性氛围中,搅拌条件下将靶向多肽滴加到步骤1)所得的酯化产物的溶液中,靶向多肽和酯化产物的摩尔比为1:(1~3),室温下反应12~48小时,经后处理得含有二硫键的靶向多肽药物偶联物。
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CN115869312B (zh) * | 2022-12-27 | 2024-02-27 | 哈尔滨吉象隆生物技术有限公司 | 一种pdc抗肿瘤药物及其制备方法与应用 |
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