CN111601622A - 用于磁共振成像诊断肿瘤的基于氧化铁磁性纳米粒子的制剂的制备方法 - Google Patents
用于磁共振成像诊断肿瘤的基于氧化铁磁性纳米粒子的制剂的制备方法 Download PDFInfo
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Abstract
本发明涉及化学领域,更具体地,涉及合成基于氧化铁磁性纳米粒子的制剂的方法,所述氧化铁磁性纳米粒子可在医学中用作肿瘤的磁共振成像(MRI)的造影剂。该方法包括以特定模式制备乙酰丙酮铁(III)的苯甲醇溶液,然后用人血清白蛋白和/或牛血清白蛋白涂覆磁性纳米粒子,并通过与戊二醛分子间交联来稳定所得涂层。磁性纳米粒子的流体动力学尺寸为25‑50nm,优选为25‑40nm,这为血液循环提供了足够的时间以进行MRI可视化。
Description
发明领域
本发明涉及化学领域,更具体地涉及氧化铁磁性纳米粒子的合成方法,氧化铁磁性纳米粒子可以在医学中用作磁共振成像(MRI)的造影剂,例如用于诊断各种肿瘤。
现有技术
当前,借助于各种造影剂可以提高对各种疾病的MRI诊断的可靠性,该造影剂可以细分为两种主要类型,其中一种包括T1造影剂,T1造影剂是含有大量未配对电子的顺磁性金属离子(Gd3+、Eu3+、Cr3+、Mn2+、Fe3+),而第二种包括T2造影剂,T2造影剂是用各种生物相容性涂层稳定的氧化铁磁性纳米粒子。
本文所要求保护的发明旨在提供以人血清白蛋白和/或牛血清白蛋白稳定的氧化铁和T2造影剂的磁性纳米粒子,其纳米粒子(或小球)的尺寸最大为50nm。T2造影剂是阴性造影剂:在MRI期间,通过磁场均匀性对信号加权,它们的积累会降低操作模式下的信号强度。重要的是,T2造影剂由于其组成的氧化铁纳米晶体而具有高了几个数量级的磁化率,其T2弛豫性能为T1造影剂的50-100倍,这降低了有效造影所需的剂量。
目前,合成磁性纳米粒子的主要方法是在表面活性剂存在下,在高沸点有机溶剂中热分解金属有机前驱体(方法1:A.H.Lu,E.L.Salabas,F.Schüth,磁性纳米粒子:合成、保护、功能化和应用(Magnetic nanoparticles:Synthesis,protection,functionalization,and application),Angew.Chemie-Int.Ed.46(2007)1222-1244,doi:10.1002/anie.200602866)以及以化学计量比共沉积金属盐(方法2:A.K.Gupta,M.Gupta,生物医学应用的氧化铁纳米粒子的合成和表面工程,生物材料(Synthesis and surfaceengineering of iron oxide nanoparticles for biomedical applications,Biomaterials),26(2005)3995–4021,doi:10.1016/j.biomaterials.2004.10.012)。方法1允许获得单分散的纳米粒子,但是表面活性剂的引入使所得的纳米粒子具有疏水性,防止其在水性介质中溶解,因此使其不适合注入人体。方法2允许获得亲水性纳米粒子,但是其具体特征是产生多相尺寸的纳米粒子的混合物,由于其组成和粒径不均匀性,阻碍了其进一步使用。
还存在其他方法,例如热解、微波合成和声化学合成,但是它们不能提供具有所需性质的氧化铁纳米粒子,使其适合MRI诊断。
磁性纳米粒子在临床实践中的使用意味着添加其具有生物相容性、无毒和可生物降解成分的制剂,以改善制剂的稳定性并降低其毒性。
但是,由于纳米粒子尺寸超过100nm,并且仅在肝脏中累积的倾向,这后一种制剂在血流量中的循环时间非常短(2-5min),这严重阻碍了其在其他器官的肿瘤诊断中的应用。
现有技术还公开了用于造影剂的磁性纳米复合材料、造影剂本身以及用于递送诊断和治疗手段的转运剂(发明申请US20130045160A1,公开日为2013年4月17日)。所述参考文献公开了使用包括白蛋白的两亲化合物涂覆纳米粒子以产生包含分布在所述两亲化合物的疏水域中的一个或多个磁性纳米粒子的磁性纳米复合材料,以及包含所述两亲化合物的亲水域的涂层,其中所述两亲化合物的所述疏水域通过物理键而非化学键与所述磁性纳米粒子的表面结合。
所述已知的纳米复合材料的缺点是,物理键与化学键相比弱几个数量级,这使得所述涂层不稳定。此外,如果在注入血液后该试剂由于疏水相互作用而被物理固定,则所述两亲化合物的涂层将被血清蛋白代替,并且两亲聚合物本身也可能被吸收在血细胞表面上,从而改变了制剂的药代动力学,并降低了其向肿瘤病灶的递送能力。
现有技术进一步公开了发明申请US20080206146A1截至2007年11月9日所包含的技术方案,该技术方案描述了磁性纳米粒子及其合成和应用。该申请公开了磁性纳米粒子,其包含提供与脑、血管和骨组织形成不同结合(conjugation)的官能团,并且意在用作MRI的诊断手段和药物转运剂。从白蛋白将功能化的磁性纳米粒子封装到核中。白蛋白的包封包括以下阶段:将200mg的人血清白蛋白溶解在2.0ml的含有磁性纳米粒子(例如磁铁矿粒子)的水中。通过在持续搅拌的同时逐滴添加0.01M和0.1M NaOH溶液,将溶液的pH调节至8.4。此外,在持续搅拌的同时逐滴添加8.0ml乙醇,从而获得10%的人血清白蛋白溶液。在添加乙醇之后,添加235μl的8%戊二醛溶液。在24小时内,通过3次循环离心(16,100g,8min)纯化所得的纳米粒子,并在水浴中通过超声处理重复进行再悬浮。涂覆的人血清白蛋白的功能化磁性纳米粒子的平均直径为60nm至990nm,具体取决于制剂的pH值和所用的粒子类型。
然而,这些纳米粒子的制备的性能特征由于大量纳米粒子的多分散性(粒径分散为60nm至990nm)而受到损害,这限制了其实际应用,因为大量的纳米粒子具有显著更短的半分泌时间,这防止了它们有效地传递到病灶。
本文公开的溶液与已知对应物的区别在于使用乙酰丙酮铁的热分解以获得具有高达20nm,优选高达12nm的初始流体动力学尺寸的单分散磁性纳米粒子,其能够溶解于水中,随后通过人/牛血清白蛋白固定化,通过戊二醛稳定化并合成包含磁性纳米粒子的制剂,该磁性纳米粒子的初始流体动力学尺寸在50nm以内,这可以消除上述对应物的典型缺点。
现有技术公开了磁性纳米粒子的合成方法(Hilda TR Wiogo,May Lim,VolgaBulmus,Jimmy Yun和Rose Amal通过胎牛血清(FBS)在生物介质中稳定磁性氧化铁纳米粒子(Stabilization of Magnetic Iron Oxide Nanoparticles in Biological Media byFetal Bovine Serum(FBS))//Langmuir 2011,Vol.27,No.2,,第843-850页),用于通过与胎牛血清混合来合成表面具有羧基的纳米粒子。上述文献公开了所述粒子的平均流体动力学尺寸为180nm,并将其在RPMI营养培养基中培育16h。上述文献还公开了蛋白质固定以非共价方式发生。但是,纳米粒子大尺寸(180nm)可能会阻止其通过血管扩散到肿瘤,从而降低其作为造影剂的效率。此外,表面的非共价修饰可能会导致注入血流后涂层解吸,从而导致与涂层结合的所有组分的损失。此外,所述参考文献没有公开任何关于评价所得造影剂用于MRI诊断的效率所需的T2弛豫性能的数据。
本文公开的本发明的最接近的对应物是MRI造影剂,其合成方法和多形性胶质母细胞瘤的MRI诊断方法(于2012年12月14日提交的专利RU2530762;Abakumov MA等人,纳米医学,2015年5月,第11卷,第4期,第825-833页)。根据所引用的文件,通过在苯甲醇中热解乙酰丙酮铁(III)合成磁性纳米粒子。在惰性气体流中,通过逐渐加热到383K,不断搅拌反应物混合物,在383K的温度下暴露1h以从溶液中蒸发水,以25K/h的速率将反应混合物的温度升高到473K,加热时间总计为9小时,并且暴露于473K的温度下持续40h,以此来合成磁性纳米粒子。在40h内,将反应混合物逐渐冷却至室温,加入90ml的无水丙酮,并通过在2000g下离心10min而与磁性纳米粒子分离。将残余物用过量的丙酮洗涤两次,并在转子蒸发器上干燥以完全除去丙酮。根据透射电子显微镜,所得纳米粒子的直径为(14±4)nm。在下一个工艺阶段,使用牛血清白蛋白(BSA)使氧化铁纳米粒子的水胶体溶液稳定。为此,向10mg粒子中加入5ml蒸馏水,并用1M NaOH溶液滴定至pH值为11。然后将所得分散体超声处理10分钟,并加入溶于5ml水中的40mg聚合物。将得到的混合物在室温下不断搅拌下培育4小时,用蒸馏水透析,然后在搅拌下逐滴加入500μl的1M NaOH,再滴加2.3ml 25%的戊二醛水溶液。将所得混合物在搅拌下培培育15min,然后加入500μl的3M甘氨酸(pH 9.2)以结合未反应的醛基。向所得溶液中加入1ml浓度为2mg/ml的氢化硼钠的PBS溶液,并培育60min。为了将牛血清白蛋白包覆的磁性纳米粒子(BSA-MNP)与多余的蛋白质分离,将纳米粒子溶液通过孔径为100kDa的纤维素离心过滤器进行过滤。将残余物重新悬浮在水中并重新过滤。重复该过程,直到洗涤液中蛋白质完全不存在为止。通过在琼脂糖凝胶(Sepharose)CL-6B(柱高50cm,直径2.5cm,流速0.7ml/min)上进行凝胶过滤,从分子交联产物中清除蛋白质。在下一阶段,将磁性粒子与流体动力学大小在150nm以内的单克隆血管内皮生长因子抗体结合,将其进一步用作MRI诊断多形性胶质母细胞瘤的造影剂。
根据RU2530762中公开的方法,在静脉内注射所得造影剂之前和之后进行MRI可视化,MRI可视化模式提供被可视化区域的磁化率加权图像,并通过在选定区域内图像亮度降低的基础上,比较在注射造影剂之前和之后拍摄的MRI图像而作出判断是否存在多形性胶质母细胞瘤。
所述方法通过增加MRI图像中对应于多形性胶质母细胞瘤组织和血管以及新血管生成病灶的区域的对比度来增加诊断的真实性和参考值。然而,根据所述方法的纳米粒子的尺寸为92nm。因此,所述方法的缺点是由于在该过程中使用长期沸腾(在200℃下40h),因此不可能获得具有小于50nm的尺寸的人血清白蛋白涂层的纳米粒子。沸腾导致纳米粒子表面的化学老化(OH基团转变成氧桥),这减少了纳米粒子表面上的带电基团的数量,从而损害了静电排斥的稳定性并降低了蛋白质吸附所需的电荷密度。此外,化学老化增加了纳米粒子的初始尺寸,这不利地影响了其在重悬浮于水介质期间的稳定性。这两种方法的结合导致长期加热后合成的纳米粒子对水介质的稳定性较差,更易于聚集。应该记住的是,源磁芯的聚集会增加已经被人血清白蛋白包覆的最终纳米粒子(小球)的尺寸。
发明内容
通过本文公开的本发明所解决的技术课题是提供一种合成血清白蛋白稳定的白蛋白磁性纳米粒子的方法,所述白蛋白磁性纳米粒子在肿瘤的MRI可视化中适合作为造影剂。
本文公开的本发明的技术结果是可以合成具有高达20nm的初始流体动力学尺寸(在HSA/BSA稳定之前)的磁性纳米粒子和具有高达50nm的HSA/BSA稳定的纳米粒子(小球),由于在初始磁性纳米粒子的合成过程中改变了乙酰丙酮铁(III)溶液在苯甲醇中沸腾的方式,因此沸腾时间在4h内。沸腾超过4小时会导致磁性纳米粒子表面发生不可逆的老化过程,从而阻碍其在水中的溶解和HSA在其表面的吸附。
使用本文要求保护的方法获得的制剂通过改善在MRI过程中注射入肿瘤中的制剂的渗透性来提高肿瘤的MRI可视化效率。这使得能够有效地穿透肿瘤组织的有缺陷的血管的孔。
通过使用MRI肿瘤诊断制剂合成方法来获得所述技术结果,该方法包括在浓度为75-200g/l苯甲醇中制备乙酰丙酮铁(III)溶液,然后在惰性气流中加热至苯甲醇沸点4-8h,并将所述溶液沸腾30min至4h,以得到悬浮液,冷却该悬浮液,用极性有机溶剂洗涤以获得Fe3O4氧化铁纳米粒子,然后用人血清白蛋白和/或牛血清白蛋白包覆,通过与戊二醛的分子间交联来稳定所得涂层。
为了用人血清白蛋白和/或牛血清白蛋白包覆,将纳米粒子溶于pH值为10-11的水中至2-8mg/ml的浓度,然后在所得溶液中加入以1:1体积比的HSA和/或BSA(即HSA和/或BSA浓度为4-16mg/ml的水溶液形式),并进行透析。
在通过分子间交联使涂层稳定之后,通过超滤和过滤灭菌再将溶液从副产物中清除。可以使用孔径为100-300kDa的过滤器实现超滤。
在本发明的优选实施方案中,以恒定速率进行加热。
所述极性有机溶剂可以选自由水溶性脂族单原子醇、丙酮、酮类和腈组成的组。所述水溶性脂族单原子醇可以选自由乙醇、甲醇或丙醇组成的组,所述酮类可以是例如丙酮或丁酮-2,所述腈可以是例如乙腈。
洗涤溶液直至除去痕量苯甲醇。在本发明的具体实施方案中,可以按照至少等于悬浮液体积的部分进行洗涤至少3个循环。另外可以通过离心沉降来实现洗涤。
使用本文要求保护的方法获得的制剂可以是溶液或冻干剂的形式。
对于肿瘤的MRI诊断,在以提供T1和T2加权的方式注射所述制剂之前,对物体(人或动物)进行MRI研究,然后以铁浓度1-10mg/kg体重的量静脉内注射制剂,然后以提供T1和T2的方式在制剂注射后2小时内再次进行MRI可视化,然后比较制剂注射前后在MR图像的可视化区域中的强度。如果确定了低强度区域,则可以得出关于肿瘤存在的结论,并概述肿瘤的范围。
本文要求保护的方法允许减少获得基于其的磁性纳米粒子和造影剂所需的时间,即带有白蛋白的磁性纳米粒子,其由尺寸小于50nm的聚集体(纳米粒子)的存在所指定。这简化了造影剂制造技术,特别是通过排除凝胶过滤阶段(与对应物技术相比)简化了造影剂制造技术。与要求长达46小时的现有技术方法相比,本文要求保护的造影剂合成方法需要5-6h至10h。此外,由于化学键合,在其他引用的著作中公开的技术不能提供与磁性纳米粒子化学键合的稳定白蛋白混合物。本发明的另一个优点是,与其他已知的在肝脏中积累的制剂不同(Majumdar S,Zoghbi SS,Gore JC.大鼠中超顺磁性氧化铁MR造影剂的药代动力学(Pharmacokinetics of superparamagnetic iron-oxide MR contrast agents in therat)Invest Radiol.1990;25:771-777),本发明要求保护的制剂在注射后数小时内通过肾脏***,因此本文避免了制剂注射的副作用。另外,本文要求保护的制剂以冻干物的形式在储存期间稳定一年,并且可以在保留所有参数的同时重新悬浮。
附图简要说明
将通过附图进一步说明本发明,其中
图1显示了使用要求保护的方法获得的氧化铁纳米粒子的显微照片,其中所述显微照片是在透射电子显微镜下获得的;
图2显示了在苯甲醇中沸腾30min后获得的磁性纳米粒子的流体动力学尺寸的数据;
图3显示了在苯甲醇中沸腾1h后获得的磁性纳米粒子的流体动力学尺寸的数据;
图4显示了在苯甲醇中沸腾4h后获得的磁性纳米粒子的流体动力学尺寸的数据;
图5显示了在苯甲醇中沸腾20h后获得的磁性纳米粒子的流体动力学尺寸的数据;
图6显示了在注射基于使用所要求保护的方法获得的氧化铁磁性纳米粒子的制剂之前大鼠C6中脑肿瘤的MRI图像(A)和在注射基于使用所要求保护的方法获得的氧化铁磁性纳米粒子的制剂之后5min的大鼠C6中脑肿瘤的MRI图像;
图7显示了在注射基于使用所要求保护的方法获得的氧化铁磁性纳米粒子的制剂之前肝脏RS-1的粘液癌的MRI图像(A)和在注射基于使用所要求保护的方法获得的氧化铁磁性纳米粒子的制剂之后5min的肝脏RS-1的粘液癌的MRI图像;
图8显示了在注射基于使用所要求保护的方法获得的氧化铁磁性纳米粒子的制剂之前小鼠乳腺癌的MRI图像(A)和在注射基于使用所要求保护的方法获得的氧化铁磁性纳米粒子的制剂之后5min的小鼠乳腺癌的MRI图像(B)。
发明实施方式
优选地,实现获得用作在肿瘤的MRI诊断中的造影剂的Fe3O4氧化铁纳米粒子的方法如下。
以下公开的是本发明的详细描述,其显示了组分含量的特定定量参数以及用于获得磁性纳米粒子的方法和用于MRI诊断的制剂的参数;然而,该描述并不将本发明的可能实施方案限制于本文中所指定的值/参数,仅旨在说明其实现提供所要求保护的结果的可能性。
为了合成,制备浓度为75-200g/l的乙酰丙酮铁(III)的苯甲醇溶液。将浓度为75-200g/l的乙酰丙酮铁(III)的苯甲醇溶液在惰性气流中加热到溶剂沸点4-8h,优选6h,再煮沸30min至4h。将反应混合物冷却至室温,离心以从溶液中分离出纳米粒子残余物,用丙酮/乙醇或甲醇、或丙醇/乙腈洗涤,并在溶剂蒸发后获得氧化铁磁性纳米粒子。
将磁性纳米粒子(80±20mg)在20ml蒸馏水中稀释,加入500μl 1MNaOH以产生碱性介质,并搅拌溶液以除去残余物。然后加入人血清白蛋白/牛血清白蛋白溶液(在20ml水中160±50mg)。将所得混合物搅拌15min,随后用0.22-0.45孔径注射过滤器过滤除去大颗粒聚集体。然后向混合物中加入500-1500μl的25%戊二醛水溶液。将反应混合物在持续搅拌下培育10-60min,加入0.5-2ml 3M甘氨酸溶液,并进一步搅拌30min至2h。然后向反应混合物中加入0.5-4ml NaBH4(10mg/ml),培育30min至4h。
过剩的低分子量物质和游离的HSA分子可通过在磷酸盐/盐缓冲溶液中在离心过滤器中洗涤除去,球形蛋白允许边缘为100-1000kDa。
如上所述获得的纳米粒子将溶解在pH值为10-11的蒸馏水中,并且当用人血清白蛋白覆盖时,它们形成由结晶磁性氧化铁核和人血清白蛋白壳组成的纳米粒子,其总尺寸在40nm以内。
用动物进行胶质瘤或其他肿瘤实验设置进行实验。为了使动物中的胶质瘤可视化,将动物置于MRI装置中,并以T2或T2*加权模式进行MR扫描。然后给动物静脉内注射HSA稳定的磁性纳米粒子溶液。注入的磁性纳米粒子剂量具有减少铁浓度为12,010mg/kg。在Bruker Clinscan 7T断层扫描仪上进行MRI扫描。静脉注射后0-24h以T2或T2*加权模式进行MRI扫描。
使用直径小于50nm的纳米粒子进行MRI诊断的优势在于,由于纳米粒子(小球)的尺寸较小,在血液中纳米粒子(小球)循环的时间增加,从而可以在不增加注射剂量的情况下实现高效可视化。此外,由于促血管生成因子的过表达(hyperexpression)导致血管网络加速生长,因此肿瘤血管具有较高的穿透性和保留能力。血管中形成的孔的尺寸为50-200nm,只有直径小于50nm的纳米粒子才能有效地穿过这些孔进入肿瘤组织。
实施例1.Fe3O4磁性纳米粒子的合成。
在搅拌的同时,将10.5g乙酰丙酮铁(III)和220g苯甲醇加热至50-70℃1h。加热速率为25℃/h。反应混合物温度达到沸点后30min停止加热。将反应混合物冷却至室温并加入90ml丙酮,并通过在900g下离心19min使纳米粒子沉降。
实施例2-5.Fe3O4磁性纳米粒子的合成。
如以上实施例1中所述合成磁性纳米粒子,不同之处在于用甲醇、乙醇、丙醇、丁醇-2或乙腈代替丙酮。
实施例6.HSA/BSA包覆的Fe3O4磁性纳米粒子的合成。
20氧化铁磁性纳米粒子中加入5ml蒸馏水,调节至pH值为11并搅拌直至完全溶解。然后在相同的pH值下加入5ml浓度为8mg/ml的HSA/BSA水溶液。将得到的混合物在室温不断搅拌下培育15min,并通过0.2-0.5μm的孔滤器过滤。向分别具有4mg/ml和2mg/ml浓度的蛋白质和Fe3+的20mg所得溶液中加入250μl的1M碱,然后逐滴加如230μl的25%戊二醛水溶液。将所得混合物在搅拌下培育15min,然后加入250μl的3M甘氨酸水溶液(pH9.2)并培育1h。然后向该溶液中加入332μl硼氢化钠的10mg/ml磷酸盐/盐缓冲溶液,并培育2h,然后将反应混合物用缓冲溶液洗涤并测量铁浓度。
实施例7.大鼠C6实验性脑肿瘤的MRI可视化。
用具有胶质瘤C6或另一类型肿瘤的实验模型的动物进行实验。为了在动物中可视化胶质瘤,将动物放置在MRI装置中,并使用以下参数在T2*加权模式下进行MR扫描:TE/TR=19/50ms,切割厚度0.5mm,FOV=30mm,分辨率256/176。然后给动物静脉内注射HSA稳定的磁性纳米粒子溶液。注射磁性纳米粒子剂量中铁离子浓度减至为5mg/kg。MRI扫描是在Bruker Clinscan 7T断层扫描仪上进行。静脉注射后5min以T2*加权模式进行MRI扫描,参数如下:TE/TR=19/50ms,切割厚度0.5mm,FOV=30mm,分辨率256/176。
在根据实施例1-5的实验过程中获得的纳米粒子的尺寸在10nm以内(图1)。
沸腾时间的增加导致纳米粒子的尺寸从7.5nm逐渐增加到50nm(图2-5)。
此外,在将磁性纳米粒子注射到具有实验性肿瘤的动物中后,随后的MRI可视化允许可视化多种类型的肿瘤,包括小鼠4T1中的脑胶质母细胞瘤C6、肝脏RS-1的粘液癌和乳腺腺癌(图6-8)。
Claims (11)
1.MRI肿瘤诊断制剂的合成方法,包括制备浓度为75-200g/l的乙酰丙酮铁(III)的苯甲醇溶液,然后在惰性气流中加热至苯甲醇沸点4-8h并煮沸所述溶液30min至4h以获得悬浮液,将所述悬浮液冷却,用极性有机溶剂洗涤以获得Fe3O4氧化铁纳米粒子,然后用人血清白蛋白和/或牛血清白蛋白包覆所述Fe3O4氧化铁纳米粒子,并通过与戊二醛分子间交联来稳定所得的涂层。
2.根据权利要求1所述的方法,其中,为了用人血清白蛋白和/或牛血清白蛋白包覆,将所述纳米粒子溶解于pH值为10-11的水中至2-8mg/ml的浓度,向所得溶液中加入水溶液形式的HSA和/或BSA,其中HSA和/或BSA体积比1:1、浓度为4-16mg/ml,然后透析所述所得溶液。
3.根据权利要求1所述的方法,其中,在通过分子间交联使涂层稳定之后,通过超滤和过滤灭菌再将所述溶液从副产物中清除。
4.根据权利要求1所述的方法,其中,可以使用孔径为100-300kDa的过滤器来实现超滤。
5.根据权利要求1所述的方法,其中,以恒定速率进行加热。
6.根据权利要求1所述的方法,其中,所述极性有机溶剂选自由水溶性脂族单原子醇、丙酮、酮类和腈组成的组。
7.根据权利要求1所述的方法,其中,所述水溶性脂族单原子醇选自由乙醇、甲醇或丙醇组成的组,所述酮类为丙酮或丁酮-2,所述腈为乙腈。
8.根据权利要求1所述的方法,其中,将所述溶液洗涤直至除去痕量苯甲醇。
9.根据权利要求8所述的方法,其中,按照部分进行洗涤至少3个循环,所述部分至少等于所述悬浮液的体积。
10.根据权利要求1所述的方法,其中,使用离心沉降来实现洗涤。
11.根据权利要求1所述的方法,其中,将使用本发明要求保护的方法获得的制剂进行冻干。
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RU2012153952A (ru) * | 2012-12-14 | 2014-07-20 | Федеральное государственное бюджетное образовательно учреждение высшего профессионального образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) | Способ диагностики мультиформной глиобластомы с помошью мрт и контрастное вещество для проведения мрт-исследования |
US20150352231A1 (en) * | 2014-06-04 | 2015-12-10 | National Taiwan University | Magnetic nanoparticle composition and manufacturing method and use thereof |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102557149A (zh) * | 2010-12-16 | 2012-07-11 | 国家纳米科学中心 | 一种制备水溶性纳米四氧化三铁的方法 |
RU2012153952A (ru) * | 2012-12-14 | 2014-07-20 | Федеральное государственное бюджетное образовательно учреждение высшего профессионального образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) | Способ диагностики мультиформной глиобластомы с помошью мрт и контрастное вещество для проведения мрт-исследования |
RU2530762C2 (ru) * | 2012-12-14 | 2014-10-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) | Способ диагностики мультиформной глиобластомы с помощью мрт |
US20150352231A1 (en) * | 2014-06-04 | 2015-12-10 | National Taiwan University | Magnetic nanoparticle composition and manufacturing method and use thereof |
Non-Patent Citations (1)
Title |
---|
ILONA S. SMOLKOVA等: "Correlation between coprecipitation reaction course and magneto-structural properties of iron oxide nanoparticles", 《MATERIALS CHEMISTRY AND PHYSICS》 * |
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