CN116162210A - 一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备方法及应用 - Google Patents
一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备方法及应用 Download PDFInfo
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Abstract
本发明公开了一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备方法,以红景天苷为模板,甲基丙烯酸(MAA)和Fe3O4@SiO2‑CC‑NH3(MCC)作为功能单体,四氢呋喃/乙二醇作为致孔剂,TRIM和二乙二醇二甲基丙烯酸酯(EGDMA)为交联剂,2,2'‑偶氮二异丁腈(AIBN)为引发剂,开发出一种具有高分子识别能力、类别特异性和选择性的缓释微球。根据紫外光谱对功能单体和红景天苷的比例进行了优化,并通过沉淀聚合法合成了一系列的MIPs;本发明公开的红景天苷分子印迹缓释剂,具有良好的吸附性能和缓释性能;对细胞生长没有任何有害影响,肠上皮细胞的存活率在98%以上。通过使用SMCMIP复合材料,药物可以以一种持续的方式递送,有利于改善治疗结果和减少副作用。
Description
技术领域
本发明涉及医药技术领域,尤其涉及一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备及应用。
背景技术
圣地红景天(Rhodiola sacra)是景天科(Crassulaceae)的植物,主要分布于西藏以及尼泊尔、锡金、不丹等地,生长于海拔3500米至5400米的林下、山坡上以及灌丛中,目前尚未进行人工引种栽培,是西藏特有珍稀药用植物,其根茎是传统藏药民间验方中的主要成分。其除酪醇、草质素、洛塞维和山奈酚等活性成分外,主要的药效成分还有红景天苷,具有抗组织缺氧、抗氧化、抗肿瘤、增强免疫、改善心血管循环、保护机体脏器、影响机体三大物质代谢和双向调节等作用,被誉为“高原人参”。
分子印迹涉及使用药物作为模板分子,在聚合物中产生选择性的结合点。磁性纳米材料具有大的比表面积、高的吸附能力和超顺磁特性,使它们易于快速分离。它们也对环境友好,制备简单,毒性低。这些特点使其在各种应用中具有吸引力。美洛昔康和5-氟尿嘧啶被用作模板分子来制备MIPs,使用Fe3O4纳米颗粒作为支架可以对其进行功能化处理。当前,磁性MIP可以用作药物递送系统,在评估其吸附机制时,确定了最大容量,并遵循Langmuir模型。使用黄蓍胶作为交联剂制备的MIP,说明了其出色识别和结合亲和力。
羧甲基壳聚糖(CC)是壳聚糖的水溶性衍生物,具有许多医疗效果,如促进伤口愈合,具有抗菌活性,可以抑制疤痕,并具有镇痛作用。正是由于其独特的生物特性,CC在生物医学和制药领域有着广泛的应用,通常用于pH值敏感的持续释放药物输送系统、环境保护和卫生保健产品。CC具有理想的特性,如生物相容性和生物降解性,使其成为伤口愈合和组织工程的有用生物材料。CC可以很容易地被加工成纳米颗粒,使其非常适用于绿色化学和药物增效输送应用。
目前,分子印迹材料(MIMs)由于其选择性结合和识别特定目标分子的能力而具有广泛的潜在应用,这包括提取、分离、纯化、催化和抗体模拟。MIMs在缓释药物递送中的应用是一个活跃的值得研究和开发的领域,关于其在该领域的应用潜力还有很多待研究。随着药学研究的系统化和科学化,开发小剂量给药、副作用小、疗效强的药物已成为一种趋势。大多数临床应用包括粗提取物粉末、片剂和颗粒,但它们有一些局限性,如爆释效应。红景天类药物上述剂型和天然产品往往不具有缓释作用;它们不能提供充分的功效,利用率低。结合现代中医治疗疾病的方法大多是针对疾病的多个方面,使用多种治疗方法,从不同角度治疗疾病,但这种治疗方式常常用时久,疗效慢。针对中医治疗疾病的弊端,如何将中药中有效成分靶向释放,首次采用磁性纳米粒子接枝羧甲基壳聚糖,设计出安全、有效、稳定的给药系统,能够快速从圣地红景天粗提物中吸附红景天苷,并用于缓释,便是本发明的初衷。
因此,本发明提供一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备及应用,以解决上述技术问题。
发明内容
有鉴于现有技术的上述缺陷,本发明所要解决的技术问题是目前现有技术中,中药有效成分提取物不具有缓释作用,药效差,利用率低的缺陷问题。
为实现上述目的,本发明第一方面提供了一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备方法,采用红景天苷为模板,MAA和Fe3O4@SiO2-CC-NH2(MCC)作为功能单体,四氢呋喃/乙二醇作为致孔剂,EDGMA为交联剂,AIBN为引发剂,沉淀得到Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹聚合物(MIPs);
进一步地,所述Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备方法,包括以下步骤:
步骤1、制备Fe3O4@SiO2-NH2:
将FeSO4·7H2O和FeCl3·6H2O分散在乙醇-氨水溶液中,并注入N2,超声混合15min。之后加入TEOS。在60℃下搅拌12小时后,将得到的纳米粒子沉淀,洗涤三次,真空干燥10h。将少量活化后的Fe3O4@SiO2微球分散在100mL甲苯中,加入APTES,在N2中室温搅拌5h。最后将Fe3O4@SiO2-NH2微球交替用二甲亚砜和乙醇洗涤三次,真空干燥12h后即得;
步骤2、Fe3O4@SiO2-CC-NH2(MCC)的制备:
将适量的CC溶于少量水中。溶解后,将Fe3O4@SiO2-NH2纳米粒子、EDC和NHS加入到CC水溶液中,超声处理30min。将该混合物缓慢滴入乳化剂Span-80和液体石蜡的混合溶液中,并在室温下搅拌反应30min。反应结束后,向混合物中加入甲醛溶液并搅拌1h。当温度上升到30℃时,缓慢加入NaOH溶液,使pH值为9.5。反应4h后,用磁铁收集MCC,并用石油醚和水清洗几次。最后,将MCC低温干燥,并计算产量。
步骤3、SMCMIP的制备
将MAA和MCC微球按照比例,加入体积比为1:1的四氢呋喃/乙二醇中,在4℃下聚合30min。以红景天苷为模板,将交联剂TRIM和EDGMA以及引发剂AIBN加入到反应体系中,并通过超声处理进行分散和溶解。用N2排除反应体系中的空气,持续10-15min。水浴振荡器的条件设定为60℃和200转,持续反应18h。用磁铁将聚合物沉淀下来。最后,用不同的溶液对聚合物进行了五次洗涤。聚合物在4℃下进行真空干燥。
进一步地,所述步骤1中,FeSO4·7H2O与FeCl3·6H2O的重量(g)比为1:1.5~1:4;
进一步地,所述步骤1中,所述FeSO4·7H2O与FeCl3·6H2O总重与乙醇-氨水溶液的重量体积比(g:mL)比为1:50:1~75;
进一步地,所述步骤2中,所述Fe3O4@SiO2-NH2纳米粒子、EDC与NHS之间的重量(g)比为10000~50000:10~18:5~10;
进一步地,所述步骤3中,所述红景天苷与MAA的摩尔比为1:4;
本发明第二方面提供了一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂;本发明提供的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂,采用上述本发明第一方面的制备方法制备得到;
本发明提供的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂,红景天苷用作模板,MAA和Fe3O4@SiO2-CC-NH2(MCC)为功能单体、四氢呋喃/乙二醇作为致孔剂,TRIM和EDGMA为交联剂,AIBN为引发剂;
本发明第三方面提供了本发明所述Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂在制备用于药物递送系统中的用途;
本发明第四方面提供了本发明所述Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂在制备用于从红景天植物中分离红景天苷的用途。
本发明第五方面提供了本发明所述Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂在制备用于从红景天植物中分离红景天苷,直接缓释施用于受药体中的用途;
具体为使用本发明的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂先从红景天植物中分离出并吸附红景天苷,然后直接将吸附有红景天苷Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂施用于受药体,进行药物治疗;所述受药体为动物或人体;
采用以上方案,本发明公开的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备方法及应用,具有以下优点:
本发明使用分子印迹聚合物(MIPs)作为药物递送系统。最近的研究表明,研发出的新型分子印迹缓释剂可以显著提高了药物的缓释性能、生物利用度和有效性。使用MIPs的药物释放可由热敏性、酸碱反应和内源性物质的存在等因素所控制。在这种情况下,Khadijeh等人制备了一种具有核壳结构的高选择性磁性MIP,它明显提高了槲皮素的最大吸附能力。在体外观察到的持续释放机制是由菲克扩散驱动的,主要是由于电化学电位梯度的作用。此外,Marcelo等人使用衣康酸制备了甲硝唑MIP的递送系统,发现MIP在pH=2.2时的释放量高于pH=7。但是,将磺胺嘧啶作为模板分子,发现口服后药物在胃中(pH=1.0)没有释放,但在肠道中(pH=6.0-8.4)缓慢释放,从而有针对性地输送到结肠。
通过对已有文献的研究,本发明成功合成了一种新型的具有核壳形态的磁性表面MIP,用于选择性识别红景天苷,以CC为磁性核心,MAA为功能性单体,MCC为支架,EGDMA为交联剂接枝的纳米粒子。其中,红景天苷的最大理论吸附量分别为130.17和370.93mg g-1。经过实验可知,本发明所得纳米粒子的磁性高亲和力、特异性结合能力以及吸附动力学速率都很高,可用于模板质量的控释。合成的SMCMIP具有良好的生物相容性,如易于制备、传质速率快、吸附能力强、易于分离且具有特定的识别能力。SMCMIP可还用于靶向药物输送,有望改善药物的治疗效果并减少副作用。
与对照组SMCNIP相比,SMCMIP复合材料在释放时间6h后具有50%的缓释特性。在25℃和37℃下,SMCMIP的释放总量分别为77%和86%。体外实验结果显示,SMCMIP的释放符合菲克动力学规律,即释放速率依赖于浓度梯度,扩散系数为3.07×10-2cm2/s~5.66×10- 3cm2/s。细胞毒性实验结果表明,SMCMIP复合材料对细胞生长没有任何有害影响。肠上皮细胞(IPEC-J2)的存活率在98%以上。通过使用SMCMIP复合材料,药物以一种持续的方式递送,有可能导致改善治疗结果和减少副作用。
以下将结合具体实施方式对本发明的构思、具体技术方案及产生的技术效果作进一步说明,以充分地了解本发明的目的、特征和效果。
附图说明
图1是实施例1得到的SMCMIP电子显微镜的扫描电镜图;
图2是实施例1得到的SMCNIP电子显微镜的扫描电镜图;
图3是试验例4的Fe3O4,Fe3O4@SiO2,Fe3O4@SiO2-NH2、CC和MCC的FT-IR光谱图;
图4是试验例4的红景天苷、SMCMIP和SMCNIP的FT-IR光谱图;
图5是试验例4的SMCMIP在室温下通过振动样品磁强计(VSM)获得的磁化曲线;
图6是试验例5的不同浓度红景天苷对SMCMIP和SMCNIP的吸附量曲线图;
图7是试验例5的红景天苷在SMCMIP和SMCNIP上的动力学曲线;
图8是试验例5的SMCMIP和SMCNIP的Scatchard图;
图9a、9b是试验例6的在模拟胃液/肠液(37℃)中,SMCMIP和SMCNIP中红景天苷的累积释放和拟合图;
图10是试验例7的SMCMIP和SMCNIP对IPEC-J2细胞生存能力的影响数据图;
图11是试验例8的红景天苷的液相色谱图;
具体实施方式
以下介绍本发明的多个优选实施例,使其技术内容更加清楚和便于理解。本发明可以通过许多不同形式的实施例来得以体现,这些实施例为示例性描述,本发明的保护范围并非仅限于文中提到的实施例。
缩写解释:
SMCMIP:Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹聚合物
SMCNIP:Fe3O4表面接枝羧甲基壳聚糖无模板分子印迹聚合物
MAA:甲基丙烯酸
MCC:Fe3O4@SiO2-CC-NH2;
TEOS:正硅酸乙酯
APTES:3-氨基丙基三甲氧基硅烷
CC:羧甲基壳聚糖
EDC:1-乙基-(3-二甲基氨基丙基)碳酰二亚胺
NHS:N-羟基琥珀酰亚胺
TRIM:三羟甲基丙烷三甲基丙烯酸酯
EDGMA:乙二醇二甲基丙烯酸酯
AIBN:偶氮二异丁腈
实施例1、Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备
(1)制备Fe3O4@SiO2-NH2
磁性Fe3O4纳米粒子是通过传统的共沉淀法制备的,Fe3O4@SiO2纳米粒子是用溶胶凝胶法制备的。具体操作为FeSO4·7H2O(0.5g)和FeCl3·6H2O(1.5g)分散在乙醇-氨水溶液(150mL)中,并注入N2,超声混合15min。之后加入TEO S(0.5mL)。在60℃下搅拌12小时后,将得到的纳米粒子沉淀,洗涤三次,真空干燥10h。将得到的Fe3O4@SiO2微球分散在100mL甲苯中,加入APTES(10mL),在N2中室温搅拌5h。最后将Fe3O4@SiO2-NH2微球交替用二甲亚砜和乙醇洗涤三次,真空干燥12h。
(2)Fe3O4@SiO2-CC-NH2(MCC)的制备
MCC是用油包水法制备的。具体操作为将适量的CC(3:4:5)溶于4.0mL水中;将Fe3O4@SiO2-NH2纳米粒子(0.01g)、EDC(10μg)和NHS(5μg)加入到CC水溶液中,超声处理30min。将该混合物缓慢滴入乳化剂Span-80(1mL)和液体石蜡(15mL)的混合溶液中,并在室温下搅拌反应30min。反应结束后,向混合物中加入甲醛溶液(1.0mL)并搅拌1h。将混合液温度上升到30℃,缓慢加入NaOH溶液(1mol/L),调整pH值为9.5。反应4h后,用磁铁收集MCC,并用石油醚和水清洗后低温干燥,得到Fe3O4@SiO2-CC-NH2(MCC)。
(3)SMCMIP的制备
将MAA和MCC微球加入体积比为1:1的四氢呋喃/乙二醇中,在4℃下聚合30min。以红景天苷为模板,红景天苷与MAA的摩尔比为1:4;将交联剂EDGMA以及引发剂AIBN加入到反应体系中,并通过超声处理进行分散和溶解。用N2排除反应体系中的空气,持续10-15min。水浴振荡器的条件设定为60℃和200转,持续反应18h。合成后,用磁铁将聚合物沉淀下来。最后,对聚合物进行了洗涤,4℃下进行真空干燥得到SMCMIP。
对实施例1得到的SMCMIP进行溶解度检测:
为了比较CMNIP和CMMIP的亲水性差异,我们研究了聚合物在水溶液中的溶胀特性。称取50~70mg的聚合物,并将其转移到一个干净的固相萃取柱中。获得总质量。在室温下使用0.9%NaCl溶液过柱,施加适当的压力以除去柱子中的过量溶液。用Kimwipe纸(Kimberly Clark Professional)擦拭MIP颗粒表面的液体后,实时监测质量,并使用以下公式(1)计算溶胀率(SR):
其中,Wt(mg)和W0(mg)分别为时间t的膨胀和原始聚合物质量;
计算得到的实施例1的SMCMIP溶胀率为67%±2%;
采用与上述实施例1相似操作,不加入红景天苷模板,其他操作相同,用同样的方法制备得到个非印迹聚合物SMCNIP。
将实施例1得到的SMCMIP,以及采用与上述实施例1相似操作得到的SMCNIP进行电子显微镜扫描,SMCMIP电子显微镜图像如图1所示;SMCNIP电子显微镜图像如图2所示;
从图1、图2对比可一看出,模板分子可在确定最终MIP产物的粒径和表面形态方面发挥作用。与SMCNIP相比,SMCMIP的表面粗糙度可能是由于模板分子的印迹,导致了更高的比表面积,改善了稳定性,对目标分析物的选择性更高(图4)。SMCMIP的形状比SMCNIP更圆,更容易分散,表明模板分子在合成过程中对球形颗粒的生长和分散有影响。SMCMIP是一种均匀的单分散颗粒,粒径约为460nm。SMCNIP的表面是光滑的,而SMCMIP的表面是粗糙的。此外,SMCMIP的平均粒径小于SMCNIP,表明印有模板分子的聚合物有一个大的表面积层,聚合物印记的聚合物被涂在SMCMIP的表面。
实施例2、
(1)制备Fe3O4@SiO2-NH2
磁性Fe3O4纳米粒子是通过传统的共沉淀法制备的,Fe3O4@SiO2纳米粒子是用溶胶凝胶法制备的。具体操作为FeSO4·7H2O(1.0g)和FeCl3·6H2O(2.0g)分散在乙醇-氨水溶液(150mL)中,并注入N2,超声混合15min。之后加入TEOS(1.0mL)。在60℃下搅拌12小时后,将得到的纳米粒子沉淀,洗涤三次,真空干燥10h。将少量活化后的Fe3O4@SiO2微球分散在100mL甲苯中,加入APTES(15mL),在N2中室温搅拌5h。最后将Fe3O4@SiO2-NH2微球交替用二甲亚砜和乙醇洗涤三次,真空干燥12h。
(2)Fe3O4@SiO2-CC-NH2(MCC)的制备
MCC是用油包水法制备的。具体操作为将适量的CC(3:4:5)溶于少量水中(5.0mL)。溶解后,将Fe3O4@SiO2-NH2纳米粒子(0.05g)、EDC(18μg)和NHS(10μg)加入到CC水溶液中,超声处理30min。将该混合物缓慢滴入乳化剂Span-80(5mL)和液体石蜡(19mL)的混合溶液中,并在室温下搅拌反应30min。反应结束后,向混合物中加入甲醛溶液(2.0mL)并搅拌1h。当温度上升到30℃时,缓慢加入NaOH溶液(1mol/L),调节pH值为9.5。反应4h后,用磁铁收集MCC,并用石油醚和水清洗后低温干燥,得到Fe3O4@SiO2-CC-NH2(MCC)。
(3)SMCMIP的制备
将MAA和MCC微球按照比例,加入体积比为1:1的四氢呋喃/乙二醇中,在4℃下聚合30min。以红景天苷为模板,红景天苷与MAA的摩尔比为1:4;将交联剂EDGMA以及引发剂AIBN加入到反应体系中,并通过超声处理进行分散和溶解。用N2排除反应体系中的空气,持续10-15min。水浴振荡器的条件设定为60℃和200转,持续反应18h。合成后,用磁铁将聚合物沉淀下来。对聚合物进行了洗涤,4℃下进行真空干燥得到SMCMIP。
实施例3、
(1)制备Fe3O4@SiO2-NH2
磁性Fe3O4纳米粒子是通过传统的共沉淀法制备的,Fe3O4@SiO2纳米粒子是用溶胶凝胶法制备的。具体操作为FeSO4·7H2O(0.6g)和FeCl3·6H2O(1.9g)分散在乙醇-氨水溶液(150mL)中,并注入N2,超声混合15min。之后加入TEOS(0.8mL)。在60℃下搅拌12小时后,将得到的纳米粒子沉淀,洗涤三次,真空干燥10h。将少量活化后的Fe3O4@SiO2微球分散在100mL甲苯中,加入APTES(13mL),在N2中室温搅拌5h。最后将Fe3O4@SiO2-NH2微球交替用二甲亚砜和乙醇洗涤三次,真空干燥12h。
(2)Fe3O4@SiO2-CC-NH2(MCC)的制备
MCC是用油包水法制备的。具体操作为将适量的CC(3:4:5)溶于少量水中(4.8mL)。溶解后,将Fe3O4@SiO2-NH2纳米粒子(0.04g)、EDC(16μg)和NHS(8μg)加入到CC水溶液中,超声处理30min。将该混合物缓慢滴入乳化剂Span-80(1~5mL)和液体石蜡(15~19mL)的混合溶液中,并在室温下搅拌反应30min。反应结束后,向混合物中加入甲醛溶液(1.0~2.0mL)并搅拌1h。当温度上升到30℃时,缓慢加入NaOH溶液(1mol/L),使pH值为9.5。反应4h后,用磁铁收集MCC,并用石油醚和水清洗后低温干燥,得到Fe3O4@SiO2-CC-NH2(MCC)。
(3)SMCMIP的制备
将MAA和MCC微球按照比例,加入体积比为1:1的四氢呋喃/乙二醇中,在4℃下聚合30min。以红景天苷为模板,红景天苷与MAA的摩尔比为1:4;将交联剂EDGMA以及引发剂AIBN加入到反应体系中,并通过超声处理进行分散和溶解。用N2排除反应体系中的空气,持续10-15min。水浴振荡器的条件设定为60℃和200转,持续反应18h。合成后,用磁铁将聚合物沉淀下来。对聚合物进行了洗涤,4℃下进行真空干燥得到SMCMIP。
实施例2、实施例3得到的SMCMIP,进行电子显微镜扫描,观察SMCMIP形态与实施例1的图1相类似;
试验例4、SMCMIP的特征检测
(1)特征吸收峰检测
对在实施例1的SMCMIP制备过程中的Fe3O4,Fe3O4@SiO2,Fe3O4@SiO2-NH2,CC和MCC,以及实施例1得到的SMCMIP,以及采用与上述实施例1相似操作得到的SMCNIP进行FT-IR光谱检测;
如图3所示,在579cm-1的特定频率的峰,是Fe-O-的特征吸收峰,而在1100.21、747.94和476cm-1,为Si-O-Si。这些峰证明SiO2成功包裹在Fe3O4纳米粒子周围。在2547cm-1,观察到的峰值归因于Fe3O4-SiO2-NH2中N-H键的扩展振动峰。同时,在FT-IR光谱中还观察到其他的峰。1670cm-1的峰值,是由C=O基团引起的。在1428cm-1的峰值,归因于-COO-基团的对称拉伸振动。在2916cm-1和3344cm-1,分别归因于-C-O-C-和-NH2基团。与CC相比,在1641cm-1,观察到一个峰值。该峰归因于C=N基团的拉伸振动,是磁性壳聚糖微球所特有的。-COO的对称拉伸振动峰,出现在1428cm-1,而-NH2和C=O峰分别出现在3344和1670cm-1。因此,表明Fe3O4磁性纳米粒子被成功地封装在CC内。
如图4所示,在FT-IR光谱中,3602cm-1和814cm-1,是盐苷的特征峰,阶梯峰在1400至1000cm-1。在1428cm-1,观察到-COO-的对称拉伸振动峰,在3334、2916和1670cm-1,可以归因于MCC,即N-H、-C-O-C-和C=O的延伸振动吸收峰。同时,在SMCMIP中也观察到了红景天苷的峰。因此,表明成功合成得到SMCMIP。
(2)磁性和吸附性检测
磁性SMCMIP具有较小的颗粒尺寸、较强的磁性和稳定的性质,使它们在各种应用中都很有用。随着壳结构的增加,磁场强度减弱并最终达到饱和状态,这表明材料的磁性能因其结构而发生变化;
如图5的VSM所示,不同级别的饱和磁力强度为70.42emu g-1。S型磁滞环、小的残余磁力和适度的矫顽力也表明,SMCMIP具有很强的磁性,并且可以长期保持其磁性。磁场使MIP均匀地分散在溶剂中并迅速被瓶壁吸收,这只需要1.5分钟。这些结果表明,SMCMIP表现出良好的磁性和有效的吸附力。
磁性MIPs具有较小的颗粒尺寸、较强的磁性和稳定的性质,使它们在各种应用中都很有用。随着壳结构的增加,磁场强度减弱并最终达到饱和状态,这表明材料的磁性能因其结构而发生变化,如VSM所示。不同级别的饱和磁力强度为70.42emu g-1。S型磁滞环、小的残余磁力和适度的矫顽力也表明,SMCMIP具有很强的磁性,并且可以长期保持其磁性。磁场使MIP均匀地分散在溶剂中并迅速被瓶壁吸收,这只需要1.5min。这些结果表明,SMCMIP表现出良好的磁性和有效的吸附力
试验例5、SMCMIP吸附实验
将实施例1得到的SMCMIP、以及采用与上述实施例1相似操作得到的SMCNIP进行吸附试验;
通过溶解5mg的聚合物(SMCMIP和SMCNIP)制备不同浓度的1mL红景天苷(0.5、1、2、3、5、7、9、10和12mg/mL),并将溶液在室温下摇晃2h。达到平衡后,进行高速离心。通过0.22μm的滤膜过滤收集上清液,并通过HPLC-UV检测。NIP以同样的方式操作。吸附量(Q,mg/mL)用公式(2)计算:
其中,
C0(mg mL-1)表示起始浓度,
Ce(mg mL-1)是平衡浓度,
V(mL)是溶液的体积,
M(mg)是聚合物质量。
其中,
Q是对SMCMIP和SMCNIP的结合能力,
Qmax是最大的结合能力,
Ce是红景天苷溶液的平衡浓度。
Kd是与吸附点的亲和力有关的解离常数。
此外,可以根据压印因子Ie,通过公式(4)对压印聚合物的选择性能进行表征,如下所示:
其中,
QSMCMIP和QSMCNIP分别是压印聚合物和非压印聚合物达到目标吸附平衡的分配系数。
结论:
为了研究聚合物的结合特性,我们在室温下测量了SMCMIP和SMCNIP在吸附达到平衡后的结合量,并说明了平衡浓度(Ce)。
如图6所示,当浓度超过5mg/mL时,SMCMIP对红景天苷的吸附量显著高于SMCNIP,但SMCMIP的吸附量较低,对存在于红景天苷中的crenulatin无选择性。Ie值为1.12。SMCMIP的吸附浓度仍然增加,表明超过一定的浓度后,SMCMIP的吸附能力主要来自于特定的结合位点。相比之下,SMCNIP的吸附涉及非特异性结合,并且是不规则的。
如图7所示,SMCMIP对红景天苷的吸附随时间增加,并在20min后达到平衡;SMCNIP在5min后达到平衡。进行了Scatchard分析,SMCMIP表现出一种特殊的吸附过程。吸附能力可主要归因于两个特定的结合点。最大的吸附能力(Qmax)值为130和371mg g-1,而SMCNIP的吸附是非特异性和不规则的。
如图8所示,对于目标分子及其类似物的结合量(Q),利用Scatchard方程研究了印迹聚合物与模板分子的结合特性。通过Q/Ce映射到红景天苷的Q,得到聚合物样品的Qmax,其中Q是吸附量;C0和Ce分别是吸附前和吸附后溶液中的样品浓度;Kd是解离常数。如图8中所示,具有明显非线性关系的两条直线表明,印迹聚合物的结合位点对于印迹分子来说并不完全等同,并且存在着两个不同的结合位点。对图中的两段线性部分进行拟合,得到了线性方程的拟合,即高亲和力和低亲和力部位方程。
试验例6、SMCMIP的体外持续释放实验
提前制得模拟胃液(pH=1.5)和肠液(pH=7.4),通过将3~8mg完全负载红景天苷的实施例1得到的SMCMIP微球浸入10mL模拟胃液(pH=1.5)和肠液(pH=7.4)中,研究了合成的SMCMIP装载药物的体外持续释放。该系统在37℃的水浴振荡器中以180rpm的速度充分振荡。在0.5~68h期间,通过离心取得1mL上清液,并加入1mL新的模拟胃液或肠液继续实验。通过HPLC-UV进行分析和检测。以同样的方式将采用与上述实施例1相似操作得到的SCMNIP作为对照实验。累积释放量(CR)用以下公式(5)计算:
其中,
W0(mg)表示吸附在聚合物上的盐苷的重量;
Cn(ng mL-1)和C(n-1)(ng mL-1)分别指在n次和n-1次所取的溶液中的盐苷浓度;
常数10.0指的是总体积,而1.0代表新鲜溶液的体积;
为了研究持续释放的机制,使用了两个模型来比较和联系一些释放参数;药物释放的动力学研究通常有助于获得有物理意义的参数。因此,使用数学模型,如Higuchi和Korsmeyer-Peppas,研究了SMCMIP和SMCNIP的扩散特性和释放动力学,这些模型用以下公式表示。
其中,
A是反映公式特征的Higuchi常数;
Mt/M∞是在时间t的一小部分的药物释放;
根据该模型,如果基质是基于扩散的药物释放机制,将Mt/M∞与t1/2进行映射将导致一条直线;
其中,
K是动力学常数,
n是描述释放机制的指数;
Mt是时间t的释放量,
K是释放率常数。
结果如下:
如图9a和9b所示,
对于SMCMIP,在模拟胃液中的释放量(在6h达到50%)比在肠液中(18h接近或低于50%)更快。然而,SMCNIP的红景天苷释放性能对pH值的变化并不敏感。在酸性pH条件下药物的快速释放率可能是由于该pH值下药物的高溶解度。由于释放介质中的氢离子必须与丙烯酸基团(pKa=4.5)竞争,因此红景天苷对SMCMIP腔的亲和力被削弱。这些因素促进了药物在酸性条件下的快速释放。在模拟的胃液中(pH=1.5),来自红景天苷和CC的聚合物羟基被电离。高离子强度克服了红景天苷和聚合物之间的静电相互作用,以促进释放。然而,药物对CC的有限性可能会产生一些离子效应,在pH=7.4的模拟液体中对基质释放过程的控制很微弱。在本组中,网状聚合物中CC的红景天苷释放率满足持续释放的要求。在25℃和37℃下对载体SMCMIP和SMCNIP进行分析和测定后,测得每个样品的提取物中的红景天苷含量分别约为77%和86%。与SMCNIP相比,37℃更有利于红景天苷的释放,其中释放率分别达到86%和77%,这可能是因为负载的SMCMIP更有利于特定的释放。因此,新型SMCMIP是连续释放盐苷的潜在候选材料。红景天苷的释放率可以用来控制药物的释放时间,从而控制应用效果。
Fick提出,扩散物质在单位时间内通过垂直于扩散方向的单位截面面积的流量与该截面的浓度梯度成正比;也就是说,浓度梯度越大,扩散流量越大。根据菲克第一定律,SMCMIP的扩散系数从3.07×10-2cm2/s到5.66×10-3cm2/s。根据SMCMIP和SMCNIP的直径与厚度的比例以及不均匀粒度分布的球形结构,n与释放指数相关。相关研究证实,对于球状结构,n<0.45;对于纯Fick扩散,0.45<n<0.89;对于Fick扩散和异常机制,n>0.89。在模拟胃液中,SMCMIP是由Fick扩散主导的。在模拟肠液中,异常扩散占主导地位。
试验例7、肠道上皮细胞(IPEC-J2)的增殖试验
细胞计数试剂盒-8用于检测细胞增殖。将处于对数生长期的IPEC-J2细胞接种到96孔板中(密度为1×106细胞/孔),在37℃,5%CO2,细胞贴壁后,加入SMCMIP(200、400、600、800、900和1000μg mL-1),持续24h。同时,建立空白组和阴性组(实施例1得到的SMCMIP和采用与上述实施例1相似操作得到的SMCNIP)。每个处理组设置六个重复。24h后,在每个孔中加入10μL CCK8溶液,在CO2中孵育2h。最后,用酶标仪在450nm处测量每个孔的OD值。
实验结果如图10所示:
生物材料的细胞毒性是衡量该材料未来应用的一个重要因素;作为安全有效的药物载体,材料本身应该对细胞无毒。因此,我们首先通过使用CCK8试验对正常的IPEC-J2细胞进行了调查。如图10所示,72h后,在SMCMIP和SMCNIP溶液中培养的细胞存活率约为98%±3%,证明SMCMIP表现出良好的生物相容性和无细胞毒性;因此,SMCMIP可以作为药物载体使用。
试验例8、红景天苷分离及施用试验
1、将实施例1制备得到的SMCMIP进行红景天植物中红景天苷成分的吸附分离,具体操作为:
(1)首先,进行磁性分离优化:
称取10mg的实施例1制备的SMCMIP样品,将其分散在10mL的丙酮中,并置于20mL离心管中。加入5mL甲醇(MeOH)和5mL CAN来激活的分子印迹聚合物,来增加SMCMIP中吸附位点的数量,去除杂质。负载溶剂用ACN、100%MeOH和纯水进行筛选。洗涤溶剂为正己烷、CCl4和CH2Cl2。洗脱溶剂分别为90%、80%和70%MeOH-乙酸。
(2)圣地红景天中红景天苷含量的SMCMIP-HPLC法测定
对10g的圣地红景天根粉末进行分离分析,浸提至水(10ML)中24h,离心10min,上清液保留。在氮气流下,上清液被还原为干燥。然后,将10mL样品稀释后,将1mL上样注入空载SMCMIP中。采用高效液相色谱法分析洗脱溶剂中红景天苷的含量。
2、然后将上述吸附有红景天苷的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂直接施用于小鼠动物,进行镇咳治疗;具体操作为:
(1)药物配制:
精准称量7.2mg上述吸附了红景天苷的SMCMIP,加入到15ml水溶液中将其分别配成1.6mg/ml、1.2mg/ml、0.8mg/ml的SMCMIP各5ml。
(2)试验动物提供:
云南昆明种小鼠,公母不限,体重20-25g。温度保持在18-25℃,相对湿度50~60%,每日的光照时间为10小时,无特殊要求下采取自由饮水、自由采食的养殖条件。饮水为自来水,食用的食物是经钴60照射灭菌饲料。小鼠的垫料在通常情况下五天进行一次更换,笼子两周进行一次清洁与消毒,饮用水每天进行添加,饲料根据小鼠情况提供。
(3)动物分组:
用小鼠作为试验动物,通过灌胃给药来研究SMCMIP对小鼠的镇咳作用。运用随机数法将试验小白鼠分为七组,第一组为空白对照组,正常饲喂,不进行任何操作。第二组为单纯引咳组,该组正常饲喂且进行引咳。第三组为溶剂组,该组在药物灌胃期灌胃PBS溶液。第四组为阳性(氢溴酸右美沙芬糖浆)组,该组在药物灌胃期灌胃10ml/kg/只的氢溴酸右美沙芬糖浆,其余与单纯引咳组一致。第五组至第七组为实验组,在药物灌胃期分别灌胃低剂40mg/kg、中剂量60mg/kg、高剂量80mg/kg的SMCMIP,其中为方便给药,将SMCMIP配制成浓度为8mg/ml、12mg/ml、16mg/ml的三种悬液,其余时期与单纯引咳组一致。
(4)实验流程:
试验总共为14天,前7天为小鼠适应期,第8-14天为试验期,分为药物灌胃与氨水引咳。
药物灌胃:经过适应期的正常饲喂七天后,进行六天的药物灌胃。对溶剂组,SMCMIP低、中、高剂量组,氢溴酸右美沙芬组进行药物灌胃。溶剂组每天7时与19时灌胃0.2mL的PBS。氢溴酸右美沙芬灌胃组及SMCMIP低、中、高剂量灌胃组每天7时与19时在灌胃前进行称重并对小鼠灌胃对应浓度、对应剂量的氢溴酸右美沙芬糖浆与SMCMIP,并观察其临床症状并进行记录。第七天上午给药前禁食不禁水12h。
氨水引咳:第七天7时对溶剂组、阳性组、实验组灌胃对应剂量,对应浓度的药品,并在1h后进行氨水引咳。在干棉球中滴加4滴浓氨水,立刻放入烧杯中,并用保鲜膜密封杯口,30s后拿出棉球,立刻把小鼠罩入,观察并记录咳嗽潜伏期以及2min内小鼠咳嗽次数(以小鼠腹肌收缩,同时张大口呼吸或小口呼吸并伴有咳嗽声记为一次咳嗽),计算咳嗽抑制率。
对上述试验数据进行统计与分析:
使用Excel软件处理测量的数据及画图,应用统计软件SPSS26.0进行样本分析统计。
试验结论如下:
1、SMCMIP对圣地红景天根提取和净化
样品在3种浓度水平(5,50,100μg kg-1)下加入红景天苷的方法对方法的准确性进行了评价。
由表1数据可知,圣地红景天根粉末中含有107.1μg kg-1的红景天苷;说明空载SMCMIP可以高效吸附根中的红景天苷。
图11显示了SMCMIP和HPLC联用后,所分离提取到的红景天苷的液相色谱图,纯度高。
综上,表明SMCMIP在红景天苷提取中的成功应用,证明了该磁性分子印迹聚合物对红景天苷选择性吸附。
表1
2.使用上述从红景天根中分离纯化,负载有红景天苷的SMCMIP直接施用于小鼠镇咳,试验数据如表2~4所示:
表2氨水引咳实验结果
分组 | n | 潜伏期 | 咳嗽次数 |
单纯引咳组 | 3 | 2.33±0.57 | 38.00±2.00 |
溶剂组 | 3 | 2.67±0.57 | 37.67±2.52 |
SMCMIP中浓度组 | 3 | 6.00±1.73 | 33.33±1.53 |
SMCMIP高浓度组 | 3 | 6.67±1.53 | 29.33±1.53 |
表4溶剂组与各组差异
由表2~4数据可知,
水溶剂组镇咳潜伏期较短(P<0.01),2min内咳嗽次数亦较多(P<0.01),说明造模成功;
与单纯引咳组的数据相比较,对照组镇咳潜伏期及咳嗽次数均无明显差别(P潜伏期=0.77,P咳嗽次数=0.87)结果无统计学差异,表明对小鼠咳嗽行为无影响;
与溶剂组比较,各用药组对氨水所致小鼠咳嗽均有抑制作用,其中SMCMIP高浓度组(P<0.01)、SMCMIP中浓度组(P<0.05)显著延长小鼠得咳嗽潜伏期;
SMCMIP高浓度组(P<0.01)、SMCMIP中浓度组(P小于0.05)的小鼠咳嗽次数明显少于单纯引咳组;
表明,本试验选用浓氨水诱发小鼠急性咳嗽,采上述负载红景天苷的SMCMIP直接施用于对氨水所致小鼠咳嗽具有抑制作用;与单纯引咳组相比,采上述负载红景天苷的SMCMIP的对氨水所致小鼠咳嗽具有明显的抑制作用。
综上所述,将本具体实施方式中实施例制备得到的SMCMIP,进行红景天根中红景天苷成分的吸附分离及纯化,然后将上述吸附有红景天苷的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂(SMCMIP)直接施用于小鼠动物,进行镇咳治疗,与对比组相比显示具有明显的咳嗽抑制作用;节省了现有技术中中药植物中有效成分在提取、分离、纯化环节的单元操作,直接用于疾病的治疗,节省了时间和成本,具有广泛的推广应用前景。
在本具体实施方式的各个试验例中,未明确注明的SMCMIP均为实施例1所制备的;未明确注明的SMCNIP均为采用与实施例1相似操作得到的SMCNIP。
以上详细描述了本发明的较佳具体实施例。应当理解,本领域的普通技术无需创造性劳动就可以根据本发明的构思做出诸多修改和变化。因此,凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的试验可以得到的技术方案,皆应在由权利要求书所确定的保护范围内。
Claims (9)
1.一种Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂的制备方法,其特征在于,采用红景天苷为模板,MAA和Fe3O4@SiO2-CC-NH3(MCC)作为功能单体,四氢呋喃/乙二醇作为致孔剂,EDGMA为交联剂,AIBN为引发剂,沉淀得到Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹聚合物(MIPs);;
其中,所述红景天苷与MAA的摩尔比为1:4。
2.如权利要求1所述制备方法,其特征在于,具体包括以下步骤:
步骤1、制备Fe3O4@SiO2-NH2:
将FeSO4-7H2O和FeCl3-6H2O分散在乙醇-氨水溶液中,并注入N2,超声混合15min。之后加入正硅酸乙酯(TEOS)。在60℃下搅拌12h后,将得到的纳米粒子沉淀,洗涤三次,真空干燥10h。将少量活化后的Fe3O4@SiO2微球分散在100mL甲苯中,加入3-氨基丙基三甲氧基硅烷(APTES),在N2中室温搅拌5h。最后将Fe3O4@SiO2-NH2微球交替用二甲亚砜和乙醇洗涤三次,真空干燥12h后即得;
步骤2、Fe3O4@SiO2-CC-NH3(MCC)的制备:
将适量的羧甲基壳聚糖(CC)溶于少量水中。溶解后,将Fe3O4@SiO2-NH2纳米粒子、1-乙基-(3-二甲基氨基丙基)碳酰二亚胺(EDC)和N-羟基琥珀酰亚胺(NHS)加入到CC水溶液中,超声处理30min。将该混合物缓慢滴入乳化剂Span-80和液体石蜡的混合溶液中,并在室温下搅拌反应30min。反应结束后,向混合物中加入甲醛溶液并搅拌1h。当温度上升到30℃时,缓慢加入NaOH溶液,使pH值为9.5。反应4h后,用磁铁收集MCC,并用石油醚和水清洗几次。最后,将MCC低温干燥,并计算产量。
步骤3、SMCMIP的制备
将MAA和MCC微球按照比例,加入体积比为1:1的四氢呋喃/乙二醇中,在4℃下聚合30min。以红景天苷为模板,将交联剂TRIM和EDGMA以及引发剂AIBN加入到反应体系中,并通过超声处理进行分散和溶解。用N2排除反应体系中的空气,持续10-15min。水浴振荡器的条件设定为60℃和200转,持续反应18h。用磁铁将聚合物沉淀下来。最后,用不同的溶液对聚合物进行了五次洗涤。聚合物在4℃下进行真空干燥。
3.如权利要求2所述制备方法,其特征在于,
所述步骤1中,FeSO4 .7H2O与FeCl3 .6H2O的重量(g)比为1:1.5~1:4。
4.如权利要求2所述制备方法,其特征在于,
所述步骤1中,所述FeSO4 .7H2O与FeCl3 .6H2O总重与乙醇-氨水溶液的重量体积比(g:mL)比为1:50~1:75。
5.如权利要求2所述制备方法,其特征在于,
所述步骤2中,所述Fe3O4@SiO2-NH2纳米粒子、EDC与NHS之间的重量(g)比为10000~50000:10~18:5~10。
6.权利要求1~5任一项所述制备方法得到的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂,其特征在于,红景天苷为模板,MAA和Fe3O4@SiO2-CC-NH3(MCC)作为功能单体,四氢呋喃/乙二醇作为致孔剂,EDGMA为交联剂,AIBN为引发剂。
7.权利要求1~5任一项所述制备方法得到的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂、权利要求6所述Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂在制备用于药物递送系统中的用途。
8.要求1~5任一项所述制备方法得到的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂、权利要求6所述Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂在制备用于从红景天植物中分离红景天苷的用途。
9.求1~5任一项所述制备方法得到的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂、权利要求6所述Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂在制备用于从红景天植物中分离红景天苷,直接缓释施用于受药体中的用途;
具体为使用本发明的Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂先从红景天植物中分离出并吸附红景天苷,然后直接将吸附有红景天苷Fe3O4表面接枝羧甲基壳聚糖红景天苷分子印迹缓释剂施用于受药体,进行药物治疗;所述受药体为动物或人体。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008025563A2 (de) * | 2006-08-31 | 2008-03-06 | Kist-Europe Forschungsgesellschaft Mbh | Polymermatrix, verfahren zu deren herstellung sowie deren verwendung |
CN104587970A (zh) * | 2014-11-24 | 2015-05-06 | 江苏大学 | 一种磁性壳聚糖复合微球表面印迹吸附剂及其制备方法 |
CN104800169A (zh) * | 2015-01-09 | 2015-07-29 | 四川大学 | 一种采用壳聚糖及β-环糊精制备疏水型药物磁靶向缓控释载体的方法 |
CN106046256A (zh) * | 2016-05-27 | 2016-10-26 | 齐鲁工业大学 | 京尼平苷分子印迹聚合物磁性微球的制备方法 |
CN113694029A (zh) * | 2021-08-02 | 2021-11-26 | 郑州大学第一附属医院 | 一种药物多孔缓释体的制备方法及药物多孔缓释体 |
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008025563A2 (de) * | 2006-08-31 | 2008-03-06 | Kist-Europe Forschungsgesellschaft Mbh | Polymermatrix, verfahren zu deren herstellung sowie deren verwendung |
CN104587970A (zh) * | 2014-11-24 | 2015-05-06 | 江苏大学 | 一种磁性壳聚糖复合微球表面印迹吸附剂及其制备方法 |
CN104800169A (zh) * | 2015-01-09 | 2015-07-29 | 四川大学 | 一种采用壳聚糖及β-环糊精制备疏水型药物磁靶向缓控释载体的方法 |
CN106046256A (zh) * | 2016-05-27 | 2016-10-26 | 齐鲁工业大学 | 京尼平苷分子印迹聚合物磁性微球的制备方法 |
CN113694029A (zh) * | 2021-08-02 | 2021-11-26 | 郑州大学第一附属医院 | 一种药物多孔缓释体的制备方法及药物多孔缓释体 |
Non-Patent Citations (6)
Title |
---|
HAILONG YU等: "Preparation of molecularly imprinted polymers coupled with high-performance liquid chromatography for the selective extraction of salidroside from Rhodiola crenulata", JOURNAL OF CHROMATOGRAPHY B, no. 1118, 16 April 2019 (2019-04-16), pages 180 - 186 * |
JIA-WEI ZHANG等: "Extraction of activated epimedium glycosides in vivo and in vitro by using bifunctional-monomer chitosan magnetic molecularly imprinted polymers and identification by UPLC-Q-TOF-MS", TALANTA, no. 219, 9 July 2020 (2020-07-09), pages 1 - 11 * |
司汇通等: "磁性分子印迹微球的合成及其对二苯并噻吩的选择性吸附研究", 功能材料, no. 1, 31 December 2016 (2016-12-31), pages 188 - 192 * |
张明磊等: "磁性Fe_3O_4@SiO_2@CS镉离子印迹聚合物的制备及吸附性能", 高等学校化学学报, vol. 32, no. 12, 31 December 2011 (2011-12-31), pages 2763 - 2768 * |
江伟等: "分子印迹吸附剂对红景天水煎液中重金属的吸附", 化工学报, vol. 59, no. 05, 31 May 2008 (2008-05-31), pages 1179 - 1183 * |
许龙等: "基于壳聚糖的分子印迹聚合物的制备和应用", 化工进展, vol. 35, no. 03, 31 December 2016 (2016-12-31), pages 847 - 855 * |
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