CN115996633A - 消毒用可分配的纳米颗粒基组合物 - Google Patents
消毒用可分配的纳米颗粒基组合物 Download PDFInfo
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- CN115996633A CN115996633A CN202180043058.2A CN202180043058A CN115996633A CN 115996633 A CN115996633 A CN 115996633A CN 202180043058 A CN202180043058 A CN 202180043058A CN 115996633 A CN115996633 A CN 115996633A
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Abstract
本文公开了快速且残留的消毒剂组合物,该组合物包括金属缔合氧化铈纳米颗粒。还公开了制造消毒剂组合物的方法。还公开了包括所公开的组合物的薄膜,以及消毒表面的方法。
Description
背景技术
由于冠状病毒易于传播,COVID-19已经给人类带来了世界性的挑战。传播被认为主要通过由感染者生产的呼吸道飞沫,以及通过接触存在含有SARS-CoV-2病毒的飞沫的表面来产生。[1]早期研究已表明,这些病毒在大多数常见类型的表面上可以存活2-3天。[2]大多数已知的可用消毒剂虽然能够中和多种类型的病毒,但通常需要大约30秒至10分钟的反应时间。[3]在这些时间范围内试图对表面进行消毒是不切实际的,这可能会引起问题。此外,目前的消毒剂需要在高接触区域内不断重复使用,因为它们不能提供残留的保护来抵御病毒和细菌。
附图说明
图1示出了应用时和应用后的RAD组合物。在右下角方框中示出了氧化铈纳米颗粒(CNP)病毒灭活的机制。
图2A示出了银改性氧化铈纳米颗粒(AgCNP)的x射线光电子能谱(XPS)测量扫描图,图2B示出了用于量化Ce3+/Ce4+比例的独特的多重态铈信号,图2C示出了详细描述AgCNP中银化学环境的银峰,图2D为银改性的CNP的高分辨透射电子显微镜(hrTEM),图2E为纯相CNP的x射线衍射。
图3示出了合成AgCNP1和AgCNP2的流程图。
图4为合成AgCNP1和AgCNP2的模型。
图5示出了AgCNP1和AgCNP2的材料特征。图5A为AgCNP1的透射电子显微镜(TEM)图像,示出了富含Ag纳米颗粒(尺寸为2-5nm)的球形颗粒(尺寸为20nm)。图5B为AgCNP2的TEM显微照片,示出了用不同尺寸的Ag纳米颗粒(5至20nm)设计的团聚的CeO2颗粒。对AgCNP1和AgCNP2进行Tafel分析,图5C示出了每种制剂的独特的腐蚀电位(分别为465.386mV和217.374mV)。图5D为10Hz至100kHz范围内AgCNP1和AgCNP2的奈奎斯特表征(Nyquistrepresation)。
图6示出了通过阻抗谱(impedance spectroscopy)原位测量AgCNP-病毒相互作用。图6A-C示出了AgCNP1与OC43包膜冠状病毒的孵育;图6D-F涉及以30分钟的有规律的时间间隔测量的AgCNP2与非包膜鼻病毒的孵育(鼻病毒和OC43病毒分别孵育总共2和4小时)。
图7为原位AgCNP-病毒相互作用的电化学模型。
图8为病毒/纳米颗粒相互作用的物理模型:
脂质体/黄嘌呤:黄嘌呤氧化酶。图8A为拟合的电化学阻抗谱,图8B示出了等效电路,图8C为拟合的电路元件值。
图9为示出了在载玻片上干燥的AgCNP2对RV14的功效的图。
图10为示出了AgCNP的重复功效的图。
具体实施方式
本文公开了一种快速且残留作用的消毒剂(ResidualActing Disinfectant,RAD)组合物,(例如,纳米RAD),以控制SARS-CoV-2和其他病原体通过与表面接触的传播。所公开的方法采用了含有快速响应的金属缔合氧化铈纳米颗粒(metal-associated ceriumoxide nanoparticles)的选择介质,其中氧化响应/机理被设计成并行的进行几个“消毒剂”反应。第一个是涉及病毒刺突糖蛋白的氧化反应,其抑制病毒-宿主细胞相互作用,从而使感染性失活。第二个机制是病毒包膜的膜过氧化以诱导溶胞(lysis);从而使病毒无效。每种消毒机制都可以通过氧化铈表面反应来实现。这些机制是自我再生的,由于纳米颗粒在消毒过程中没有用完,从而允许纳米RAD具有残留的消毒能力。在其他实施方案中,颗粒可以通过掺入银变得更有效:导致在应用中进一步生产自由基。用氟或类似的化学成分掺杂纳米氧化铈,可以将前两个机制的反应速率降低到远远低于30秒。消毒机制的组合(共同作用)将进一步减少总比率事件,允许通过多种并行途径进行快速的消毒,并在安全接触的浓度下进行干燥的消毒功效。
根据一个实施方案,公开了一种可分配的组合物,其包括金属缔合氧化铈纳米颗粒(mCNP)和赋形剂。与氧化铈纳米颗粒缔合的金属可以包括但不限于银、金、钌、钒、铜、钛、镍、铂、钛、锡和铁。在一个具体的实施例中,金属为银,并且占颗粒的重量的10%或者更少。在一些实施方案中,赋形剂选自水、氯仿、二氯甲烷、丙酮、甲基乙基酮、环己烷、乙酸乙酯、乙醚、低级醇、低级二醇、THF、DMSO或DMF。mCNP还可以掺杂有氟。
在其他实施方案中,公开了生产mCNP的方法。其中金属为银时,通过以下方法生产AgCNP,所述方法包括:溶解铈前体盐和银前体盐,如硝酸铈和硝酸银;通过含有过氧化物的掺杂剂(admixture)氧化已溶解的铈前体盐和银前体盐;和通过含有氢氧化铵的掺杂剂沉淀纳米颗粒。或者,通过包括以下步骤的方法生产AgCNP:(i)溶解铈前体盐和银前体盐,如硝酸铈和硝酸银;(ii)通过含有氢氧化铵的掺杂剂氧化和沉淀已溶解的铈前体盐和银前体盐;(iii)在水中洗涤和再悬浮沉淀的纳米颗粒;(iv)用过氧化氢处理再悬浮的纳米颗粒;和(v)洗涤来自步骤(iv)的纳米颗粒以除去离子化的银。
在其他的实施方案中,公开了通过将可分配的组合物实施方案分配到表面上来消毒表面的方法。下面将进一步描述这些和其他实施方案。
定义
除非具体规定或者从上下文中显而易见,否则如本文中所使用的,术语“约”被理解为在本领域中的正常公差范围内,例如在平均值的2个标准偏差内。约可以被理解为在规定值的30%、25%、20%、15%、10%、9%、8%、7%、6%、5%、4%、3%、2%、1%、0.5%、0.1%、0.05%或0.01%之内。除非上下文中另有说明,否则本文提供的所有数值都用术语“约”来修饰。
本文中所使用的术语“消毒(disinfection)”或“消毒(disinfect)”是指减少或消除表面上的病原微生物(包括细菌和病毒)。本文中所使用的术语“残留的消毒剂”是指能够以干燥的形式对表面消毒至少24小时的任何喷洒的消毒剂。持续长达24小时的残留的消毒剂可以在10分钟内减少3log病毒载量和减少5log细菌载量进行消毒。持续时间超过一天的残留的消毒剂(喷洒或通过其他方式应用)可以在2小时内以减少3log病毒载量和减少3log细菌载量进行消毒。
本文中所使用的术语“快速的消毒”是指几乎瞬间消除表面上的病原微生物。当以湿的形式应用时,快速的消毒剂具有约1分钟或更少的消毒停留时间。
术语“金属缔合氧化铈(cerium oxide)纳米颗粒”、“金属缔合氧化铈(ceria)纳米颗粒”或“mCNP”是指掺杂有或以其他方式结合到金属(如银、金、铜、铂、镍、铁、钛、钌、钒等)的氧化铈纳米颗粒。术语mCNP包括AgCNP。在一个实施方案中,金属缔合氧化铈纳米颗粒包括1nm至50nm,或5nm至100nm,或5nm至25nm的范围的颗粒尺寸。
本文中所使用的术语“纳米RAD”是指含有与金属(如银)结合的氧化铈纳米颗粒作为活性剂和赋形剂的消毒剂。如本文所教导的,所公开的纳米RAD组合物可以包括赋形剂(如有机酸)、表面活性剂、干燥剂和/或聚合物等。
本文中所使用的术语“分配”通常是指从容器或分配系统中喷出组合物。分配可以例如通过使用空气交换泵、开口等来完成。本文对分配组合物的量或方式没有限制。在某些实施方案中,组合物可以以类似于雾化的喷雾的细小雾状物的形式进行分配,这可以通过使用例如喷嘴或雾化器来实现。在其它实施方案中,组合物可以在高压或低压下,以单股液体流(液滴等)的形式进行分配。在本发明的实施方案中,可以利用满足特定环境需要的任何形式的分配。
本文中所使用的术语“泵”是指能够分配位于容器内的组合物的装置。泵可以是“空气交换”泵,其通过将空气或类似物注入容器中来起作用。然后,注入的空气置换并分配容器内的部分或全部的组合物。分配的组合物的量取决于注入的空气的量和容器内组合物的量。更具体地,泵可以将空气注入容器中,并将组合物从喷嘴或其他开口中分配出来。
术语“占主导的4+表面电荷”是指表面上铈离子的浓度,并意味着氧化铈纳米颗粒表面上的[Ce3+]:[Ce4+]比小于50%。在一个具体的实施例中,具有占主导的4+表面电荷的氧化铈纳米颗粒具有40%或更小的[Ce3+]:[Ce4+]比。
术语“占主导的3+表面电荷”是指氧化铈纳米颗粒表面上的[Ce3+]:[Ce4+]比大于50%。在一个具体的实施例中,[Ce3+]:[Ce4+]的比率大于60%。
术语“湿化学合成”是指一种制造CNP的方法,该方法包括在水中溶解铈前体盐,然后加入过氧化氢。在一个具体的实施例中,CNP在预定的时间内保持稳定,通常至少15-30天。
综述
目前的消毒剂喷雾仅在应用时消毒。应用后,所公开的RAD组合物具有独特的能力,能够在其应用的表面上产生临时的、持续的消毒薄膜。持续的消毒活性是由于氧化铈纳米颗粒(CNP)纳米表面反应位点的再生(催化)性能,当新的病毒或细菌接触表面时,该性能允许对表面进行持续消毒。对于不易应用永久性消毒薄膜的表面,这提供了一种有吸引力的解决方案。RAD组合物是一种解决方案,其能够通过目前不可获得的且作为消毒剂喷雾和临时薄膜独特的方式控制与表面接触的COVID 19和医院获得性感染(HAIs)的传播。
随着COVID-19的出现,许多企业和政府已在努力解决如何让人们以减少冠状病毒传播的方式进入公共场所或公共空间。在印度,当购物者进入市场时,穿行式喷灌系统已被用来直接向市场购物者喷洒消毒剂。[4]由于冠状病毒的高度传播性,许多人甚至在益处尚不清楚的情况下争相地寻找控制传播的解决方案。
冠状病毒和许多呼吸道病毒一样,通过呼吸道飞沫传播。这意味着当人们出现在一个区域时,打喷嚏、说话和咳嗽会将呼吸道飞沫沉积在表面上。在正常的表面上,通过使用商业上可获得的消毒剂喷雾,这些飞沫将保留已经以稳定的形式嵌入其中的任何病毒,直到应用消毒剂喷雾,或者在应用消毒剂喷雾一段时间之后(可能长达2至3天)。人们研究了永久性抗病毒薄膜,以帮助控制SARS-CoV-2的传播。永久性薄膜对其应用的表面有特定的粘附要求,以防止分层。此外,这些薄膜主要旨在防止表面润湿,作为对抗病毒传播的间接措施,而不是直接灭活病毒物种。与目前可获得的表面消毒能力相比,RAD组合物具有能够使表面保持更长时间的消毒能力。永久性消毒薄膜很难改进到现有的表面上,并且可能需要更换/修改部件或材料来提供它们的益处。当商业上可购买时,RAD组合物将结合商业上可获得的喷雾和薄膜的优点,提供几乎没有持久性的喷雾的急性消毒能力和永久性薄膜的一些优点。
疾病控制中心(CDC)通过国家儿童保健和早期教育健康与安全资源中心制定了儿童保育设施表面消毒的指南。[6]建议的消毒计划表包括使用前、使用后和每日(每天结束时)的指南,表1。应该注意的是,这个推荐的计划表是从CDC网站的COVID-19日托设施指南上链接的。[7]该表选择的是经常接触的可能有助于冠状病毒传播的表面。其中许多仅建议在一天结束时进行清洁。鉴于SARS-CoV-2的高感染性,以及许多人无症状但却是病毒的携带者,这些清洁措施是不够的。它们为某人在表面附近打喷嚏、咳嗽或说话提供机会,并在从未实际身体上接触表面的情况下沉积呼吸飞沫。然而,应用RAD组合物以延长应用后的消毒时间将使该消毒计划表在防止病毒通过表面传播方面更加可靠。
表1:清洁、杀菌(Sanitizing)和消毒的常规计划表(改编自[6])
不同于其它可获得的表面消毒剂,所公开的RAD组合物提供了目前表面消毒剂不具备的能力:临时的、持续的消毒薄膜。对于负责冠状病毒传播高风险场所的消费者来说,这一特征将使RAD组合物成为有吸引力的替代解决方案。
在一个实施方案中,提供了快速作用的消毒剂(RAD)喷雾,其通过与受污染的表面接触控制病毒(例如,SARS-CoV-2)的传播。RAD喷雾采用了一种含有快速反应的掺杂CNP的选择介质,其氧化反应被设计为并行的进行多种消毒机制(表2)。图1示出了RAD组合物如何工作以对抗像冠状病毒活性氧(ROS)物质这样的呼吸道病毒的操作概念,其是与其他直接CNP表面反应机制(膜过氧化和S蛋白氧化)一起使用的机制之一,以提高消毒速率以及每个单独CNP的消毒效率。[14]消毒机制的组合(共同作用)提高了总消毒率,允许通过多种并行途径进行快速且有效的消毒。应用后,所公开的RAD组合物具有独特的能力,能够在其应用的表面上产生临时的、持续的消毒薄膜。CNP具有再生特性,当来自呼吸道飞沫或物理传播的新病毒遇到它时,该再生特性允许表面的持续消毒。对于不易应用永久性消毒薄膜的表面,这提供了一种有吸引力的解决方案,允许应用到多种类型的表面上,而不考虑表面粘附薄膜的能力。在一个具体的实施例中,RAD组合物是一种解决方案,其能够通过目前不可获得的且作为消毒剂喷雾和临时薄膜独特的方式控制与表面接触的COVID 19和其它病原体的传播。在本文中将更详细地讨论这些机制。
表2:纳米RAD是一种快速作用、残留的消毒剂喷雾,在最初被应用并对表面进行消毒后,持续安全消毒数天。
目前,CNP已作为广谱抗病毒剂在体外实验式地使用。由于其独特的化学(例如,增强的催化活性)性质,它们被用作预防病毒感染的替代方法。据推测,当纳米颗粒被生物流体(例如呼吸道飞沫)水化时,表面氧化还原反应产生ROS,并伴随氧化应激,诱导病毒包膜的脂质过氧化,影响病毒的稳定性,导致表面受体蛋白氧化,从而使病毒丧失感染性(即通过修饰受体来阻止宿主细胞与病毒的相互作用)。
不同类型的纳米颗粒已被证明是抗病毒剂,如金、银和氧化铈。其中,CNP对正常型(normo-typic)细胞具有最小毒性或无毒性,并调节氧化还原相关的细胞过程,使细胞存活或死亡,并根据合成程序对氧代谢物质表现出独特的催化活性。氧化铈可以以两种形式存在:1)Ce2O3具有六边形[27]和2)CeO2具有立方萤石晶格。这赋予了纳米氧化铈以下性质:氧储存和释放,催化作用[27,28]和太阳能/燃料电池。[29]
就CNP而言,氧空位的产生导致两个电子在4f态上的定位。[27,30,31]这导致了具有热力学稳定结构的两个配位铈阳离子(从Ce4+到Ce3+)/氧气的还原。[27,31]此外,在纳米水平上,纳米氧化铈中的可获得的表面积和晶体平面的取向高度调节催化性能。以前已经证明,在原子密度最高的晶体平面中,由于其相对较高的原子间间距,纳米氧化铈的(100)平面族[32]表现出最高的反应性[33]这在以前是通过改变纳米氧化铈颗粒的形貌来说明的,这可以通过改变制备的合成方法来控制,并在不同的晶体平面确定马德隆能(Madelungenergies)[34]。
这些氧空位成为催化活性的位点,并随颗粒尺寸而变化。[35]CNP具有不同的取决于它们的表面化学的酶-模拟活性。由于+4表面氧化态的存在,过氧化氢酶模拟活性高,而超氧化物歧化酶活性由于更多的Ce3+而增加。此外,CNP(Ce3+至Ce4)中的这些混合价态具有在晶体系统内部在氧化态之间切换的能力。当改变其价态时,CNP可以清除活性氧物质(ROS)和活性氮物质(RNS)。在生物系统中,通过助氧化剂和抗氧化剂发生重要的生物和环境反应。助氧化剂通过产生羟基自由基(OH)、过氧化氢(H2O2)和超氧阴离子(O2 -)来诱导氧化应激(其可能使病毒破坏)。在正常和癌变条件下,催化CNP通过氧化还原反应被用于减少人体各器官中的活性氧物质。[18,38-40]
CNP被用作抗微生物剂[41]和抗病毒剂。[42]纳米氧化铈通过直接作用于细菌结构或间接地通过化学改性来充当抗生素剂。CNP可以直接与细菌细胞壁相互作用,从而导致细胞壁不稳定和溶胞。或者,颗粒可以间接作用;与细胞内的化学物质和成分反应。每种机制都会导致细菌细胞死亡。在生理pH值下,CNP上的正电荷导致基于这些机制的抵御细菌物质的抗微生物活性,由最初的膜粘附介导。[43,44]就病毒而言,CNP的几何形状和表面电荷作为抗病毒剂起着重要作用。在细胞渗透/病毒摄取之前,通过将CNP连接到病毒表面,可以改变独特的生物化学性质和病毒驱动的(virus-motivated)细胞间级联反应。Lozovski等人证明了窄的、小尺寸的CNP分布对含有DNA和RNA的病毒具有最显著的效果。[42,45]这是由于释放的离子引起磷酸酶模拟活性的局部效应,以及干扰钙依赖的膜过程。此外,这些离子物质被证明可以调节代谢过程,特别是在线粒体处或附近(例如电子传递链事件)。[46]CNP容易附着在磷酸盐基团上,导致无机、不溶性磷酸铈。[47]此外,CNP已被证明可以加速核酸中高抗性磷酸二酯键的断裂。[46]当CNP与细胞表面蛋白相互作用时,会导致细胞表面性质的改变。这些可能包括膜的胶体性质及其流动性,从而影响病毒进入活细胞的能力。特别设计的纳米氧化铈,无论有无银掺杂,都是全面抗病毒治疗和灭活新兴的COVID-19和其他病毒和病原体造成的表面污染的候选物。
实施方案的描述
本发明描述了掺杂或以其他方式与金属(如银、金、铜、铂、镍、铁、钛、钌、钒等)结合的氧化铈纳米颗粒。已经在多种抗菌/抗病毒应用中研究了金属和金属氧化物纳米材料的使用,并为病原体毒性提供了更广泛的基础。过渡金属基材料已经显示出优异的广谱抗菌活性以及抗病毒功效。
mCNP可以为球形、杆状、星形或多边形。在一个优选的实施方案中,mCNP是球形的,这意味着它们或多或少近似于球形。优选地,球形mCNP的平均直径为约24nm或更小,约20nm至约24nm或约3nm至约5nm。在某个实施方案中,通过透射电子显微镜测得球形氧化铈纳米颗粒的平均直径为3nm至5nm。在mCNP不是球形的实施方案中,优选纳米颗粒的两个相对侧之间的平均尺寸为24nm或更小。
mCNP具有氧化铈核心,该核心具有外表面。根据其上Ce(3+)离子相对于Ce(4+)离子的百分比来表征该表面。虽然不旨在限制用量,但当在本发明的方法中使用时,Ce(3+):Ce(4+)百分比的一些优选范围是:约80%:20%至约20%:80%、约75%:25%至约25%:75%、约60%:40%至约25%:75%、或约57%:43%至约27%:73%。在某些实施方案中,Ce(3+)相对于Ce(4+)的百分比>50%Ce(3+)。
银缔合氧化铈纳米颗粒
本公开包括两种不同类型的纳米颗粒AgCNP1和AgCNP2。在某些实施方案中,存在两者的组合,因为它们似乎有略微不同的作用模式。银改性氧化铈制剂(AgCNP)是以两种独特的制剂(AgCNP1,AgCNP2)合成的,每种制剂利用了针对银水溶液的不同化学反应。AgCNP1是通过先前开发的两步法(图3A、图4)合成的,可以被缩放到大或小的方法(process)。简而言之,通过碱性强制水解反应形成含有类似AgCNP(AgCNP-like)、银改性纳米氧化铈和银次生相(silver secondaryphases)的溶液。用去离子水(dH2O)洗涤产物材料,随后用氢氧化铵处理。氢氧化铵起到蚀刻剂以及相转移络合物的作用:介导水相中溶解的银离子的溶解/稳定。特别地,该反应导致形成托伦试剂(Ag[(NH3)2OH]aq)。然后用dH2O洗涤所得的单颗粒溶液,以去除过量的碱和反离子/旁观离子。AgCNP2利用了银离子对过氧化氢氧化的稳定性(图3B)。具体而言,溶解硝酸铈和硝酸银,随后加入过氧化氢导致铈离子选择性氧化银,并在氧化铈表面演变成金属银相。这些颗粒的独特的合成条件表明了潜在的完全不同的颗粒特性。在某些实施方案中,合成可以被缩放到大或小的方法。
AgCNP2的小规模方法的实施例:
1.将109mg六水合硝酸铈(99.999%,纯度)溶解在50mL方形玻璃底部的47.75mL,dH2O中。
2.将250μL,0.2MAgNO3(99%,纯度)水溶液添加到上述铈溶液中,并使溶液涡旋2分钟:机器:旋涡器(Vortexer)。
3.从这里,将2mL,3%过氧化氢(储备)快速加入到上述溶液中,随后立即以最高转速涡旋2分钟(在涡旋器机器中)。
4.溶液在室温、黑暗条件下储存,并松开瓶(50mL方底玻璃杯)盖,以允许释放逐渐形成的气体;在这些条件下,溶液老化长达3周(监测溶液颜色由黄色变至澄清),以产生50mL总体积的溶液。
5.然后将颗粒用2L dH2O透析2天以上(透析管),每12小时换一次水,并在与老化相同的条件下储存。
氧化铈纳米颗粒的两种独特的制剂是用由银纳米相改性的表面生产的。材料表征示出每种制剂中的银组分彼此互不相同,并装饰氧化铈表面作为许多小纳米晶体(AgCNP1)或作为Janus-型两相结构(AgCNP2)。优选地,AgCNP1的平均直径为约20至24nm,AgCNP2的平均直径为约3至5nm。每个合成还具有独特的混合的原子价,相对于AgCNP2上的Ce4+,AgCNP2具有显著更大比例的Ce3+态。独特的原子价特征,以及化学活性银相的结合,导致每种制剂的高催化活性。AgCNP2具有高超氧化物歧化酶活性,而AgCNP1同时具有过氧化氢酶和类超氧化物歧化酶模拟酶活性,这归因于氧化铈的过氧化氢酶活性和来自银相的超氧化物歧化酶活性。此外,电化学分析表明,与纯银相相比,掺入每个制剂中的银对氧化还原介导的降解基本上更稳定:促进了催化应用中寿命的增加。每种制剂在影响抗病毒特性中的用途示出了每种制剂的特定活性:在被测试的病毒物种中,AgCNP1示出了对OC43冠状病毒的显著活性,AgCNP2示出了对RV14鼻病毒的活性。在各自的孵育期内为每种病毒/颗粒系统收集的原位电化学阻抗谱反映了对每个配对观察到的独特的相互作用。每个等效电路配件,以及开发的模型/测试系统(使用模拟病毒样颗粒、模型蛋白、氧自由基物质生成酶/底物系统),示出了配对在影响抗病毒反应中的作用模式。这些研究的结果为OC43/AgCNP1确定了主要的基于物理相互作用的机制,而RV14/AgCNP2确定了氧化、化学相互作用。
虽然不旨在限制用量,但当在本发明的方法中使用时,一些与AgCNP有关的银百分比的优选量为约6%至约10%或更少。
组合物的实施
在一个实施方案中,提供了包括mCNP(例如AgCNP)和赋形剂的可分配的组合物。赋形剂的实例包括溶剂,例如但不限于水或水基(水性)溶液(其中水至少是主要成分)、低级醇(C6或更低级),低级二醇(C6或更低级)、THF、DMSO、DMF等。它们可以单独使用或作为各种组分与水的混合物使用。不构成对非水性载体或其混合物的限制的实例是氯仿、二氯甲烷、丙酮、甲基乙基酮、环己烷、乙酸乙酯、乙醚、低级醇(C4或更少)、低级二醇(C4或更少)、THF、DMSO和DMF。
可分配的组合物也可以包括香料。香料的实例包括但不限于柠檬油、橙油、佛手柑油、依兰油、广藿香油、香茅油、柠檬草油、boad玫瑰油、丁香油、桉树油、雪松油、薰衣草油、天然香料如檀香油、香根草油、香叶油、赖百当油、薄荷油、玫瑰油、茉莉花油、利兹阿克贝巴油(litz accubeba oil);烃类香料(例如柠檬烯、α-蒎烯、莰烯、对伞花烃、phen Chen等)、醚类香料(例如,1,8-桉叶素、玫瑰氧化物、雪松醇甲醚(cedlum bar)、对甲酚甲醚、异戊基苯基乙醚、4-苯基-2,4,6-三甲基-1,3-二氧六环、茴香脑等)、S类香料(例如,乙酸乙酯、丙酸乙酯、丁酸甲酯、异丁酸乙酯、丁酸乙酯、乙酸丁酯、2-甲基丁酸乙酯、乙酸异戊酯、2-甲基戊酸乙酯(锰酸盐))、乙酸己酯、己酸烯丙酯、三环癸烯基丙酸酯(VERTOPRO;氟环烯)、庚酸烯丙酯、乙酸异冰片酯、乙酸芳樟酯、乙酸香茅酯、2-叔丁基环己基乙酸酯(narcidol)等)、醇类香料(例如,亚麻醇、3-辛醇、2,6-二甲基-庚醇、10-十一烯醇(10-undecenol)、香叶醇、橙花醇、香茅醇、松香油、mill Senol、四氢芳樟醇、麝香草酚、松油醇、雪松醇、2,4-二甲基-3-环己烷-1-甲醇、4-异丙基环己醇、橙花叔醇、9-癸烯醇、顺-3-己烯醇、反-2-己烯醇、丁香酚等)、醛类香料(例如,香茅醛、仲醛、苯甲醛、醛C-6、醛C-7、醛C-8、醛C-9、醛C-10、tripral、对乙基二甲基氢化桂皮醛等)、合成香料(如海风醛(florazone)、2-十三烯醛、醛C11等)或与这些共混的共混香料。
根据其他的实施方案,如本文中所教导的,基底可以涂有金属缔合氧化铈纳米颗粒的薄膜。基底可以采取在其上进行人体接触或者通常放置人体呼出的飞沫的任何表面的形式,如纸巾、卫生纸、工作台面、HVAC过滤器、空气净化装置、电风扇、冰箱、微波炉、洗碗机/烘干机、电饭煲、锅、锅盖、IH加热器、洗衣机、真空吸尘器、照明设备(灯、设备主体、灯罩等)、卫生用品、马桶、洗脸盆、镜子、浴室(墙壁、天花板、地板等)、建筑材料(内墙、天花板材料、地板、外墙等)、室内产品(窗帘、地毯、桌子、椅子、沙发、架子、床、床上用品等)、眼镜、腰带、扶手、门、旋钮、衣服、家用电器或类似用品的过滤器等、文具、厨具、医疗用品(白大褂、口罩、手套等)、医疗器械和设备、以及在汽车、火车、飞机、小船和轮船等内部使用的材料。基底材料的实例包括玻璃、陶瓷、塑料、树脂(如丙烯酸树脂)、纸、纤维、金属、木材等。
在另一个实施方案中,还可以生产被用于多种应用的抗病毒泡沫。例如,使用通过将异氰酸酯与多元醇(具有三个或更多个羟基的分子)、扩链剂(双官能羟基分子)、促进反应的催化剂、表面活性剂、热和/或UV稳定剂以及发泡剂混合而产生的制剂来制造聚氨酯泡沫。发泡剂可以为水,因为当它与异氰酸酯反应时会产生二氧化碳气体。制造抗病毒泡沫的方法包括用表面活性剂(使用与系统相容的表面活性剂或与系统中使用的相同的表面活性剂)或氨基甲酸酯形成组分之一生产金属缔合氧化铈纳米颗粒,并将这些添加到泡沫制剂中。另一种替代方法包括在水性介质中生产纳米颗粒,例如通过将它们与所需的表面活性剂在水中混合,然后将这种水性混合物作为发泡剂和抗病毒源添加到泡沫制剂中。
根据其他实施方案,可以使用印刷墨水领域中已知的技术形成包含与银或另一种金属结合的氧化铈纳米颗粒的抗病毒墨水。可以使用多种技术(例如喷墨印刷、苯胺印刷、凹版印刷和丝网印刷)印刷这种墨水。在某些情况下,例如在喷墨印刷中,官能化颗粒的尺寸应该小于约50nm。可以通过3-D打印形成三维抗病毒产品(口罩材料和通常接触的硬物体),其中3-D打印组合物包含本文中教导的抗病毒材料,例如AgCNP。
喷雾制剂
本发明还包括纳米RAD的喷雾制剂。在典型的实施方案中,制剂包括纳米RAD、干燥剂、有机酸、表面活性剂、水和聚合物粘合剂。在某些实施方案中,根据所需的消毒机制,纳米RAD可以包含一种或几种mCNP。当应用于基底时,纳米RAD喷雾生产消毒薄膜。在某些实施方案中,纳米RAD的量为约0.01至10重量%。在某些实施方案中,干燥剂(如乙醇或异丙醇)的量为约0至40重量%。在某些实施方案中,向喷雾制剂中提供约0.5至2重量%的柠檬酸或其他有机酸。其他干燥剂包括醇或醇的混合物,例如乙醇、异丙醇、正丙醇及其混合物;脂肪醇,包括但不限于鲸蜡醇、肉豆蔻醇、硬脂醇、辛醇、癸醇和月桂醇,及其混合物;己醇和/或其他脂肪族或芳香族醇。可以用于所公开的组合物的有机酸包括但不限于乳酸、柠檬酸、水杨酸、乙醇酸、扁桃酸、苯甲酸及其组合。
纳米RAD还可以与根据应用途径选择的相容的表面活性剂、稀释剂和聚合物粘合剂混合。表面活性剂可以作为洗涤剂、湿润剂、乳化剂、起泡剂或分散剂。在某些实施方案中,表面活性剂的量为约0.5至3重量%。合适的表面活性剂为例如,月桂基胺氧化物、肉豆蔻胺氧化物、其他两性离子剂、tergitol 15-S-15或其他仲醇乙氧基化物。在某些实施方案中,月桂基胺氧化物的量为约0.25至2重量%,tergitol 15-S-15的量为约0至1重量%。在某些实施方案中,合适的稀释剂为水,其量为约15至45重量%。聚合物粘合剂用于生产透明、柔韧、透氧的薄膜,该薄膜粘附在玻璃、塑料和金属上。合适的聚合物粘合剂例如,聚(2-乙基-2-噁唑啉)或聚乙烯吡咯烷酮(PVP)-乙酸乙烯酯共聚物。在某些实施方案中,PVP-乙酸乙烯酯共聚物的量为约1至30重量%。在某些实施方案中,聚(2-乙基-2-噁唑啉)的量为约1至25重量%。
适用于所公开的组合物的其它聚合物包括聚环氧乙烷(Polyox)水凝胶聚合物、硬脂醇、纤维素聚合物、阳离子羟乙基纤维素(例如,Ucare;JR30)、羟丙基甲基纤维素、羟丙基纤维素(Klucel)、壳聚糖吡咯烷酮羧酸盐(Kytamer)、山嵛醇、硬脂酸锌、乳化蜡(包括但不限于Incroquat和Polawax)、丙烯酸的加成聚合物、树脂(如ETD 2020)、瓜尔胶、阿拉伯树胶、丙烯酸酯/硬脂醚-20甲基丙烯酸酯共聚物、琼脂、藻胶、藻酸、丙烯酸铵共聚物、藻酸铵、氯化铵、硫酸铵、支链淀粉、凹凸棒石、膨润土、C9-15醇、醋酸钙、藻酸钙、卡拉胶钙、氯化钙、辛醇、卡波姆910、卡波姆934、卡波姆940P、卡波姆940、卡波姆941、羧甲基羟乙基纤维素、羧甲基羟丙基瓜尔胶、卡拉胶、纤维素、纤维素胶、鲸蜡硬脂醇、鲸蜡醇、玉米淀粉、达玛树脂、糊精、二联苯胺山梨醇、乙烯二氢化牛油酰胺、乙烯二酰胺、乙烯二硬脂酸酰胺、明胶、瓜尔胶、瓜尔胶羟丙基三甲基氯化铵、锂蒙脱石、透明质酸、水合二氧化硅、羟丁基甲基纤维素、羟乙基纤维素、羟基乙基乙基纤维素、羟乙基硬脂酰胺-MIPA、异鲸蜡醇、异硬脂醇、刺梧桐树胶、海带、月桂醇、豆胶、硅酸镁铝、硅酸镁、三硅酸镁、甲氧基聚乙二醇(PEG)-22/十二烷基乙二醇共聚物、甲基纤维素、微晶纤维素、蒙脱土、肉豆蔻醇、燕麦粉、油醇、棕榈仁醇、果胶、PEG-2M、PEG-5M、聚丙烯酸、聚乙烯醇、海藻酸钾、聚丙烯酸铝钾、卡拉胶钾、氯化钾、硫酸钾、马铃薯淀粉、藻酸丙二醇酯、丙烯酸钠/乙烯醇共聚物、羧甲基葡聚糖钠、卡拉胶钠、纤维素硫酸钠、氯化钠、聚甲基丙烯酸钠、硅铝酸钠、硫酸钠、司拉氯铵膨润土(stearalkonium bentotnite)、司拉氯铵水辉石(stearalkonium hectorite)、硬脂醇、牛油醇、三乙胺盐酸盐、黄蓍胶、十三醇、三甲胺硅酸镁铝、小麦粉、小麦淀粉、黄原胶、松香醇、丙烯酸亚油酸、山嵛酸铝、辛酸铝、二聚亚油酸铝盐(如二硬脂酸铝和异硬脂酸铝)、蜂蜡、山嵛酸酰胺、丁二烯/丙烯腈共聚物、C29-70酸、山嵛酸钙、硬脂酸钙、小烛树蜡、棕榈蜡、地蜡、胆固醇、胆固醇羟基硬脂酸酯、椰子醇、柯巴脂、二硬脂酸甘油酯苹果酸酯、二氢枞醇、二甲基月桂基胺油酸盐、月桂酸/鲸蜡硬脂醇/乙二醇共聚物、芥酸酰胺、乙基纤维素、三乙酰羟基硬脂酸甘油酯、三乙酰蓖麻酸甘油酯、乙二醇二山嵛酸酯、二-辛酸乙二醇酯、乙二醇二硬脂酸酯、己二醇二硬脂酸酯、氢化C6-14烯烃聚合物、氢化蓖麻油、氢化棉籽油、氢化猪油、氢化鲱油、氢化棕榈仁油甘油脂类、氢化棕榈仁油、氢化棕榈油、氢化聚异丁烯、氢化大豆油、氢化牛油酰胺、氢化牛油甘油酯、氢化植物油甘油酯、氢化植物油、日本蜡、荷荷巴蜡、羊毛脂醇、乳木果油、月桂酰胺、脱氢松香酸甲酯、氢化松香酸甲酯、松香酸甲酯、甲基苯乙烯/乙烯基甲苯共聚物、微晶蜡、蒙脱土酸蜡、蒙脱土蜡、十四烷二十醇、肉豆寇十八醇(myristyloctadecanol)、十八烯/马来酸酐共聚物、十八烷基硬脂酰硬脂酸酯、油酰胺、油硬脂(oleostearine)、小冠椰子蜡、氧化聚乙烯、地蜡、石蜡、戊赤藓醇氢化松香酸酯、戊赤藓醇四辛酸酯、戊赤藓醇松香酸酯、戊赤藓醇四松香酸酯、戊赤藓醇四山嵛酸酯、戊赤藓醇四油酸酯、季戊四醇硬脂酸酯、眼用酸酐/甘油/癸酸缩水甘油酯共聚物、眼用/偏苯三酸酐/乙二醇共聚物、聚丁烯、聚对苯二甲酸丁二醇酯、聚二戊烯、聚乙烯,聚异丁烯、聚异戊二烯、聚乙烯醇缩丁醛、聚乙烯醇月桂酸酯、二癸酸酯、二元酸丙二醇酯、二异烟酸丙二醇酯、二月桂酸丙二醇酯、二壬酸丙二醇酯、丙烯乙二醇二硬脂酸酯、二十一酸丙二醇酯、PVP/二十烯(eiconsene)共聚物、PVP/十六碳烯共聚物、米糠蜡、硬脂酸铵膨润土、十八烷基铵锂蒙脱石(水辉石)、硬脂酰胺、硬脂酰胺DEA-二硬脂酸酯、硬脂酰胺DIBA-硬脂酸酯、硬脂酰胺MEA-硬脂酸酯、硬脂酮、硬脂酰芥酸酰胺、硬脂酸十八酯、硬脂酰硬脂酸甘油酯、合成蜂蜡、合成蜡、三羟基硬脂酸、三异壬精(triisononanoin)、三异硬脂酸、三异硬脂酸三亚油酸酯、甘油三月桂酸酯、三亚油酸、三亚油酸甘油酯、三肉豆蔻酸甘油酯、三油酸甘油酯、软脂酸甘油酯、三硬脂精、月桂酸锌、肉豆蔻酸锌、新癸酸锌(zinc neodecanoate)、松香酸锌及其混合物。用于车辆的胶凝剂可以是天然胶凝剂,如天然胶、淀粉、果胶、琼脂和明胶,可以基于多糖或蛋白质。实例包括但不限于瓜尔胶、黄多糖胶、藻酸(E400)、海藻酸钠(E401)、海藻酸钾(E402)、海藻酸铵(E403)、海藻酸钙(E404-褐藻多糖)、琼脂(E406,从红藻中获得的一种多糖)、卡拉胶(E407,从红藻中获得的一种多糖)、刺槐豆胶(E410,来源于树莓的种子的一种天然树胶)、果胶(E440,从苹果或柑橘类水果中获得的一种多糖)和明胶(E441,由动物胶原蛋白部分水解而成)、1,5-戊二醇4-t-烯基环己醇(Symsitive 1609)。
泵喷洒组合物实施例:
·0.01-5%重量的纳米RAD(活性)
·0-40%重量的乙醇(亚异丙醇-或其他干燥剂)
·0.5-2%重量的柠檬酸
·0.5-3%表面活性剂
·0.25-2%月桂胺氧化物(亚肉豆蔻胺氧化物-或其他两性离子剂)
·0-1%tergitol 15-S-15(非离子表面活性剂:仲醇乙氧基化物)
·15-45%水
·1-25%聚(2-乙基-2-噁唑啉)(聚合物粘合剂)或类似聚合物
在某些实施方案中,纳米RAD喷雾制剂在应用时产生可再水化的薄膜,并在再水化时显示潜在的持续消毒行为。AgCNP可以从气态水颗粒中提取水用于再活化,由喷雾制剂产生的聚合物薄膜也是亲水性的,这有助于实现气态水颗粒表面水层用于再活化消毒行为。
根据其它实施方案,提供了一种容器,该容器具有用于分配本文中所述组合物的泵。泵可以以任何方式设计,该方式满足组合物和容器的限制,并且以期望的方式分配组合物。此外,泵可以包括延伸到容器中的管,从而促进泵分配液体的能力。本领域技术人员将会理解,包括可选的管、喷嘴等的泵可以与容器内的组合物流体连通。泵也可以设计成“可拆卸地耦合”到容器上,这意味着它可以从容器上拆卸和重新连接一次或多次。
另一个实施方案涉及一种装置,该装置包括用于容纳一定量的本文中公开的可分配的组合物的容器部分和喷嘴。在具体实施方案中,该设备包括适于容纳组合物的容器;以及耦合到容器的泵,该泵包括喷嘴并且与组合物流体连通,该泵被配置为通过将空气注入容器以置换组合物来从喷嘴分配组合物。在具体实施方案中,泵还包括延伸到容器中并与组合物流体连通的管子。
在另一个实施方案中,该装置包括用推进剂加压的流体密封的容器和在被启动时分配可分配的组合物的阀。本领域非常熟知的用于分散组合物的合适的推进剂。常见的推进剂的实例包括但不限于碳氢化合物、乙醚、压缩气体、含氯氟烃推进剂、液体推进剂或其混合物。
根据本文中的教导可以使用的分配容器类型的一些实例包括但不限于美国专利号3061202、美国专利号3986644、美国专利号4669664、美国专利号5358179、美国专利号3995778、美国专利号4202470、美国专利号3992003、中国专利号1042213、美国专利公开号20180370715、美国专利号2863699和美国专利号3333743的专利中公开的装置类型。
生物相容性和安全
意识到对纳米氧化铈毒理学的关注,已经对铈盐的反应性进行了研究,这项工作激发了对氧化铈纳米毒理学的兴趣[21]。另一项研究考察了CNP表面电荷和尺寸的变化以及对细胞摄取的影响。[48]此外,利用CNP的荧光共轭物进行了另一项研究,分析了纳米氧化铈的动力学和亚细胞定位。[49]由于裸露的氧化物纳米材料在哺乳动物中的生物相容性可能不如软材料,因此进行了一项专注于PEG功能化的研究,以确定聚乙二醇化是否会改变CNP的催化性质,结果并没有。[50]
合成纳米氧化铈颗粒的方法有多种,包括湿化学法、溶剂热法、微乳液法、沉淀法、水解法和水热法。[51,52]根据所采用的合成方法,这些纳米颗粒的尺寸从3-5nm到100nm以上不等,表面电荷从-57mV到+45mV不等。合成方法也会影响CNP的形状。涂层和表面活性剂也可能存在并污染制剂,如环六亚甲基四胺(HMT)[53]或乙二醇。[54]许多报告纳米氧化铈毒性的研究着眼于通过水热法产生的NPs。这种类型的CNP通常具有会对细胞造成伤害的尖锐边缘。[55]然而,一种湿化学制剂合成的CNP生物相容性更好,且几乎零毒性。在人脐静脉内皮细胞(HUVECs)中观察到了这种毒性的缺乏。[56]
虽然它是无毒的中性pH正常型细胞,但由于酸性化学环境和纳米氧化铈的pH敏感氧化还原活性,它在杀死癌细胞方面仍然非常有效[57]。还观察到了这些CNP之前报道的CNP的保护作用。在一篇综述文章中,38份报告示出了CNP在细胞培养和动物研究中的保护作用。[51]应该注意的是,许多细胞类型和动物模型已经暴露于纳米氧化铈,并显示出有益的效果。这些细胞包括RAW 264.7巨噬细胞、BEAS-2B肺细胞、H9c2心肌细胞、A549肺细胞、HT22海马神经细胞、器官型神经元和许多其他。动物模型包括Tubby突变小鼠、EAE模型、C57BL/6小鼠、糖尿病Wistar大鼠和异位肿瘤小鼠模型。
实施例
实施例1:纯相和银改性氧化铈纳米颗粒诱导模拟生物流体中ROS的制剂。
COVID-19和其他类似流感病毒对人类健康构成了实质性威胁,因为它们通过受感染个体释放的生物液体具有高度感染性。由于在高流量地区(highly trafficked area)与受污染的表面接触,人对人的感染在第一反应和医疗环境中尤其明显。目前的消毒措施要么在这些环境中不可用,要么由于机械动力学的限制而显示出有限的功效。研究表明,纳米氧化铈和银-纳米氧化铈将在高反应速率下表现出ROS诱导作用,这是由于在满载病毒的生物流体存在下的纳米尺度/表面效应。产生的ROS引起大量氧化应激从而导致膜过氧化和溶胞,以及引起病毒细胞受体表面结构的氧化和失活从而导致病毒失活。关于纳米相银和氧化铈的文献提出了在相关条件(请参阅FIG1)下推测的ROS生成反应方案。
2Ag(0)+O2+H2O→Ag2O+O2 -+2H+
2Ce3++Ag2O+2H+→2Ce4++2Ag(0)+H2O/自由基引发
(i)O2 -+Ag2O+2H+→H2O2+Ag2O/表面扩散;超氧化物组合
(ii)O2 -+2H2O+Ce+→H2O2+Ce4+/基于氧化铈的SOD模拟活性
(iii)O2 -+Ce3++2H+→H2O2+Ce4+/氧化铈SOD模拟活性
AgOx+H2O2→Ag++H2O+H+/过氧化物介导的氧化溶解
1.1通过各种基于溶液的途径合成颗粒,并进行初步表征。鉴于化学环境对纳米材料表面化学的强烈影响以及纳米医学文献中证明的促进独特ROS生成的氧化铈氧化还原比(即涉及Ce3+和Ce4+分数(fraction)的相对材料组成)的影响,研究了几种合成方法。纯相纳米氧化铈是通过几种独特的方法合成的,这些方法在之前被证明可以诱导ROS的产生。在一个实施例中,基于过氧化氢的氧化反应被用于生产高Ce3+/Ce4+比例的纳米氧化铈制剂。特别是,将硝酸铈六水合物在水中溶解至5mM,然后在搅拌下加入3%的过氧化氢。将颗粒静置一段时间,以允许氧化铈表面降解过量的过氧化物。
为了生产更富含Ce4+的制剂,利用强制水解方法进行第二次合成。具体而言,颗粒是由六水合硝酸铈前体在水溶液中形成的。过氧化氢限制了金属相和氧化银相的形成(即,防止次要的、不同的银纳米相形成)。因此,一些合成将利用过氧化物作为银改性纳米氧化铈制剂中的氧化剂。首先,通过原位方法生产制剂,其中溶解硝酸铈和硝酸银,然后直接过氧化氢氧化,并老化以允许通过氧化铈表面催化来降解过氧化物。其次,进行混合强制水解方法,其中将溶解的盐首先通过过氧化物氧化,随后通过加入30%氢氧化铵来沉淀。通过以10,000rpm离心收集颗粒,并用去离子水洗涤三次。直接过氧化物介导的氧化和强制水解方法的结合将介导铈氧化还原态比例的变化。第三,制备溶液,其中共溶解的硝酸铈和硝酸银进行氢氧化铵介导的氧化/沉淀,随后洗涤和再悬浮在去离子水中。从这里,加入过氧化氢,并在搅拌下静置溶液,以促进次生相银纳米材料的溶解。随后洗涤颗粒以除去离子化的银。通过过氧化物或氢氧化铵的氧化经由独特的化学反应形成氧化物颗粒,从而强烈影响产物银-纳米氧化铈。在每个纳米材料候选制剂中考察了银分数(质量百分比;2%、5%、10%、20%)的影响。颗粒尺寸和表面电荷通过动态光散射和ζ-电位测量进行评估。此外,银相特征和Ce3+/Ce4+分别通过监测约320nm和252/298nm处的峰进行定性评估(图4)。
1.2对1.1中生成的制剂进行ROS生成化学活性分析。特别地,使用标准的生物测定试剂盒评估过氧化氢酶和超氧化物歧化酶的活性。羟基自由基生成活性通过分析加入的亚甲蓝染料的降解来评估。在模型生物流体溶液中进行化验(例如pH值为6的NaCl/HCl缓冲液、室温)。收集并比较与每个反应相关的反应速率。评估了银释放/电离在这些反应中的影响。电离反应首先通过在恒定的时间点的UV-Vis测量(即分析银离子峰演变)进行监测,随后通过光谱电化学(即监测UV-Vis峰特征,同时在开路电位下进行电流分析法和伏安法/Tafel分析,以详细说明银腐蚀过程)进行监测。此外,通过滴定和Tafel分析来测定氯化物浓度对反应(速率)的影响。使用商业脂质过氧化测定试剂盒(MDA测定)分析纳米材料诱导脂质过氧化的功效。这些研究的综合结果(collective result)被用于修改1.1中的合成参数,以生成引发ROS产生的高反应速率的Ag-CNP制剂。
实施例2:表征纳米颗粒并分析功效和毒性。
关于CNP的初步工作已经证明了何种形式的CNP导致不同类型的生物行为。据显示,CNP和Ag-CNP将产生ROS,这将使包膜病毒的磷脂双层失活-这导致这类病毒的快速和广泛的溶胞和失活,使它们不能感染细胞。
2.1从尺寸、形态和化学成分方面表征展现高ROS生成反应速率的制剂。高分辨率透射电子显微镜(hrTEM;以展现纳米材料尺寸、形态和颗粒特征)、小角x射线衍射(SAXS;以表征银和氧化铈相结晶特征)、以及x射线光电子能谱(XPS;以分析/评估化学成分、铈氧化还原比和银氧化/化学环境,如图2所示)。
2.2CNP和Ag-CNP通过蚀斑试验(plaque assay)或TCID50试验从悬浮溶液感染的病毒物质中评估降低感染性。从这里,RT-PCR被用于分析病毒基因组。为每种制剂建立了病毒灭活的剂量依赖性和时间依赖性。
采用两种方法来确定CNP和Ag-CNP灭活一系列人类致病病毒的能力。首先,将不同浓度的病毒在固定浓度的CNP或Ag-CNP的溶液中孵育。在混合后的不同时间,从样品中取出试样,稀释并分析剩余的感染性。是否使用蚀斑试验或TCID50取决于病毒。使用实时PCR来确定剩余的颗粒(不考虑感染性)。样品被分成三份进行分析,数据被表示为与我们先前出版物中所示的病毒的起始水平相比,感染性倍数变化。[61,62]温度是病毒稳定性的主要因素,并与孵育的时间和CNP或Ag-CNP的浓度一起测试。
首先用冠状病毒的原型实验室毒株进行上述化验,以便可以实现快速的进展和显示生产力。为了确定CNP的抗病毒特异性,测试病毒#2(寨卡病毒,Zika virus)以确定具有与CoV相似结构的其他包膜正指向(positive-sense)RNA病毒是否也对灭活敏感。病毒#3(鼻病毒)测试敏感性是否延伸到缺少脂质双层的正指向RNA病毒。病毒#4(甲型流感病毒)测试包膜负指向RNA病毒的敏感性,该结果将对作用机制产生影响。病毒#5(痘苗病毒VV)测试含DNA包膜病毒的灭活。根据已发表的示出了VV比RNA病毒对化学处理的抵抗力更强的工作成果[61](Bracey等人,2019),预计会出现敏感性梯度-CoV>流感>VV。非包膜鼻病毒的结果将是重要的,因为这将指导关于灭活是脂质依赖性还是核酸依赖性的未来研究。
如果观察到任何包膜RNA病毒(例如冠状病毒、寨卡病毒、流感病毒)失活,则表明包膜已被CNP或Ag-CNP破坏。这涉及到样品的蔗糖梯度沉降,该样品包括单独的CNP、单独的病毒和如上确定的孵育的CNP加病毒。离心后,收集组分并通过蛋白质印迹分析病毒组分的位置。完整的病毒沉淀到试管的底部附近,而破碎的病毒体则保留在梯度的顶部。CNP与病毒体的直接相互作用通过交联实验和测试CNP与颗粒共沉淀的梯度分数来检测。
实施例3:优化的银改性氧化铈纳米颗粒气雾剂和载体组分的制剂。
气雾剂或泵喷雾介导的消毒剂的分散允许快速、广泛地部署到一般表面上,而不对材料特性或拓扑结构进行显著考虑。将Ag-CNP包含到气雾剂或喷雾剂中,可作为一种便携式系统,用于一般表面的消毒,具有高消毒率,干燥后仍有持续残留的消毒活性。此外,这种纳米材料在气雾剂介质中的储存将有助于纳米材料活性成分的长期保存;从而在给药前保持活性。
3.1:Ag-CNP分散在不同挥发性的溶剂(例如,醇类、醚类)中。根据颗粒制备方法,分散或者通过在洗涤步骤后将颗粒悬浮在候选分散剂中来完成,或者通过透析去除水相来完成。通过动态光散射(即相对于hrTEM测量,随着尺寸的变化,测量颗粒的溶解动力学半径(solvo-dynamic radii)和聚集特性)和ζ电位(表面溶剂配位影响稳定性;ζ电位>25mV被认为是高度稳定的)评估胶体稳定性。可以添加无害的配体物质(例如非反应性的小、极性有机物质,如糖类),通过协调颗粒表面赋予更好的稳定性。最佳的分散剂(或推进剂)是基于更大的挥发性(从而在喷洒后通过含病毒的生物流体介导有效的水合作用)和纳米颗粒胶体稳定性。
3.2将Ag-CNP悬浮在分散剂介质中,并在生物流体模型溶液中稀释。ROS生成通过分析随时间的变化来监测,以估计载体介质汽化过程中的功效。相对于纯模型介质中的活性来比较反应速率。
实施例4:用于表面和薄膜性能的制剂的优化。
由制剂溶液形成临时薄膜的不同方法是可能的。这些方法包括由制剂悬浮液形成弱膜、Ag-CNP对表面的范德华粘附以及NPs对表面的弱静电相互作用。制剂的活性组分的小微晶性质将允许基于一种或多种这些机制形成临时薄膜。
4.1喷雾制剂在满载病毒的表面上的功效和干燥制剂在施用病毒/生物流体时作为薄膜的功效。将测试制剂喷洒在接种过病毒的测试表面,以确定初始功效。通过将颗粒分散在测试表面上,随后接种病毒并测定相互作用后的病毒的感染性,来分析喷雾作为(干燥的)薄膜的功效。
将薄膜孵育不同的时间,感染并如上所述对感染性和总颗粒进行处理。将AgCNP2应用到载玻片上,并允许干燥1小时。将Rhino14送至AgCNP2处理的载玻片和未处理的载玻片上。在两个小时的过程中,AgCNP2载玻片上的病毒滴度以显著高于未处理载玻片的速率下降(图9)。AgCNP1和AgCNP 2分别针对OC43和RV14的残留功效分析显示,AgCNP在数小时内保持其功效(图10)。
实施例5:金属介导的纳米氧化铈通过表面破坏灭活人冠状病毒和鼻病毒的优化。
在本研究中,生产、表征了两种独特的银改性氧化铈纳米颗粒制剂(AgCNP1、AgCNP2),并测试了抗病毒功效(图5)。显微镜和光电子能谱示出了在铈的氧化还原态组成、制剂颗粒尺寸和氧化铈基体中银相存在方面存在明显差异。电化学和带隙测量提供了对银和银/氧化铈界面性质的了解,并提供了它们被氧化铈相稳定的证据。通过一组独特的病毒类型测定抗病毒功效,AgCNP制剂在其抗病毒活性中示出对特定病毒的特异性。本文中,测定并比较了对鼻病毒RV14和冠状病毒OC43的抗病毒功效。第一次,进行了原位电化学阻抗谱方法,并证实了AgCNP制剂/病毒类型在孵育期内相互作用的特异性。根据该数据,以及设计的类似系统的结果,确定了用于描述高效病毒/AgCNP制剂对的抗病毒活性的作用机制的一般模式。
5.1材料合成和胶体特性:
银改性氧化铈制剂(AgCNP)是以两种独特的制剂(AgCNP1,AgCNP2)合成的,每种制剂都利用了针对银的水溶液的不同化学反应。AgCNP1是通过先前开发的两步法(图5)合成的。简而言之,通过碱性强制水解反应形成含有类似AgCNP、银改性纳米氧化铈和银次生相的溶液。用dH2O洗涤产物材料,随后用氢氧化铵处理。氢氧化铵起到蚀刻剂和相转移络合物的作用:介导水相中溶解的银离子的溶解/稳定。特别地,该反应导致形成托伦试剂(Ag[(NH3)2OH]aq)。然后用dH2O洗涤所得的单颗粒溶液,以除去过量的碱和反离子/旁观离子。AgCNP2利用了银离子对过氧化氢氧化的稳定性。具体而言,溶解硝酸铈和硝酸银,然后加入过氧化氢,导致铈离子选择性氧化银,并在氧化铈表面演变成金属银相。这些颗粒的独特合成条件表明了潜在的完全不同的颗粒特性。
对颗粒的胶体特性进行了动力学稳定性、表面电位和流体动力学直径的评估。在表3中收集每个样品的动态光散射(DLS)测量结果,与每种制剂的更大粒径(包括特定的水合球体)相关,其中AgCNP1颗粒的直径约3倍大(表3)。此外,ζ电位测量表明AgCNP2比AgCNP1具有更大的表面电位,每个都具有正极性。这些特征描述表明观察到AgCNP2颗粒示出比AgCNP1更大的动力学稳定性(AgCNP1:在室温条件下颗粒老化1周后适度沉淀;AgCNP2:超过5个月没有明显的沉淀)。AgCNP1颗粒在1mg/mL的溶液中也表现为浑浊,而AgCNP2在类似条件下完全半透明,表明更大的Mie散射与更大的粒径有关。观察每种合成的颗粒,以证明独特的基本材料特性和功能材料特性。
表3.AgCNP制剂的物理化学性质。
5.3电化学表征
XPS结果表明每种制剂具有独特的银特性,因此,通过常见的电化学技术评价每种制剂中银相的稳定性。通过电化学测量(图5C,D)以确定AgCNP制剂中银相的活性及其对电子转移过程的敏感性。在XPS结果的证实中,AgCNP1被证实具有比AgCNP2更大的Tafel电位(表3)(分别为465.4mV对217.4mV),表明对电子转移更强的稳定性和更高的氧化特性。有趣的是,AgCNP1显示了Tafel电流,其是AgCNP2观察到的值的两倍(分别为0.027μA和0.013μA)。这些值相对较低,表明每种制剂中银相的总体稳定性。然而,可以从XPS光谱了解到AgCNP1在较高电位下的较大电流值,其中发现一部分样品中的银含量为氧化物。银渗透到氧化铈表面/亚表面将增大Tafel电位(即,对银相具有稳定作用),同时改善相在其界面的入驻(registry),改善作为Tafel电流的电荷转移。TEM图像证实AgCNP1中的银-氧化铈比AgCNP2中的银-氧化铈具有更大的界面面积。在Tafel分析(表3)中观察到两个样品的阳极β值明显大于阴极β值,表明氧化过程在动力学上有利于Tafel电位下的电子转移。虽然电化学方法可以提供关于基本电荷转移过程的信息,但这种表征仅提供处于原子水平或化学水平上的名义信息。
5.5用AgCNP1和AgCNP2选择性灭活两种人类呼吸道病毒:
为了确定纳米氧化铈和银改性纳米氧化铈能够灭活人类冠状病毒OC43的程度,制备反应以包括每毫升105病毒感染单位(TCID50)以及缓冲液和纳米颗粒。或者,单独的缓冲液反应包括水作为载体对照。105TCID50/mL输入病毒被确定为时间零感染性。孵育6小时后,单独的缓冲液对照反应具有104TCID50/mL剩余感染性病毒。未改性纳米氧化铈,CNP2和CNP1,对病毒滴度几乎没有影响,反应保持在约5×104TCID50/mL。引人注目的是,AgCNP1处理导致感染性病毒完全失活,而AgCNP2处理使得感染性病毒滴度降低至约103TCID50/mL。用如上所述制备的仅包含缓冲液、AgCNP1或AgCNP2的反应进行时间进程研究。在孵育0、2、4和6小时后确定感染性病毒。早在4小时,AgCNP1处理就使OC43病毒滴度从初始值105TCID50/mL降低至小于102TCID50/mL。总之,这些数据表明AgCNP1在灭活冠状病毒OC43方面非常有效,AgCNP2具有适度的灭活OC43的能力。
为了确定灭活冠状病毒OC43的最佳有效AgCNP1浓度,从105TCID50/mL OC43开始,连同缓冲液和递增浓度的AgCNP1一起制备反应。在孵育5分钟和4小时后确定感染性病毒。在5分钟时间点后,所有的AgCNP1浓度(约为104-105TCID50/mL)具有相似的病毒滴度。相比之下,用0.77mg/mLAgCNP1处理4小时,结果是没有检测到OC43病毒感染性,用0.2mg/mLAgCNP1处理,将感染性降低至约102TCID50/mL。AgCNP1-OC43灭活的结果用另一种感染性的方法来证实。104蚀斑形成单位(PFU)/mLOC43用单独的缓冲液孵育4小时,恢复了所有的感染性,与0.77mg/mLAgCNP1孵育相比,其结果是在化验中没有检测到OC43蚀斑。总之,这些数据示出了AgCNP1对灭活冠状病毒OC43感染性的时间依赖性和剂量依赖性。
接下来我们试图确定纳米氧化铈和银改性纳米氧化铈能够灭活人类呼吸道病原体鼻病毒14(RV14),非包膜的二十面体(icosahedral)RNA病毒的程度。RV14用单独的缓冲液或所示的纳米颗粒孵育。用水作为载体对照制备单独的缓冲液反应。测定6×105TCID50/mL输入RV14病毒,并表示为时间零点。孵育6小时后,单独的缓冲液反应保留了6×105TCID50/mL的输入感染性。未改性纳米氧化铈CNP2和CNP1对RV14的感染性几乎没有影响。重要的是,AgCNP1处理使得感染性病毒滴度降低至5×102TCID50/mL,而AgCNP2处理导致感染性病毒完全失活。在时间进程研究中,6×105TCID50/mL的RV14用单独的缓冲液孵育6小时后,没有示出感染性的丧失。相比之下,通过用AgCNP2孵育2小时,RV14的感染性非常迅速地降低至检测不到的水平。与AgCNP2相比,用AgCNP1孵育显示RV14的灭活较慢,6小时后病毒滴度降低至约102TCID50/mL。总之,这些数据表明AgCNP1和AgCNP2都可以灭活RV14的感染性,其中AgCNP2具有更强的抗RV14效果。
5.6AgCNP消毒剂活性的原位生物电化学阻抗谱表征:
这两种制剂对病毒物种的独特亚群表现出基本的消毒活性,以及HA化验结果表明了在每种测试案例下独特的作用模式。为了探究每种制剂的特性,对两种测试案例,即AgCNP1/OC43和AgCNP2/鼻病毒(图6)进行了电化学阻抗谱(EIS)。EIS是一种非破坏性表征技术,它依赖于在固定范围内变化的频率下施加小振幅电位。将测得的电流分解成来自独特频率区域的贡献,允许确定特征电化学过程。EIS是制造业中的主要技术,尤其是对于能源和半导体工业。在本文中,总阻抗是用简单电路图(即,用代表化学成分/过程的拟合电路元件)拟合的数据测量的。近年来,该技术已被应用于研究物理或化学刺激对细胞特性的改变。在这些研究中,最常研究的是细胞膜特性改变的条件。Giaever和Keese的ECIS模型给出了细胞-基质EIS数据的简单解释,其中阻抗成分(impedance components)被解卷积为生物颗粒之间以及颗粒和电极基质之间区域的电荷流的电阻和细胞膜电容。在细胞健康的研究中,这些模型成分是诊断性的:每种成分在引入毒性剂后都会发生变化(例如,膜孔形成、病灶区域收缩、膜氧化)。此外,这些反应必然是频率依赖的,该频率具有被独特的生物过程识别的特定频带。特别突出显示三个区域,分别表示为α(<10kHz)、β(10kHz<100mHz)和γ(GHz)。在测试剂(例如AgCNP)存在的情况下识别阻抗谱随时间的变化时,可以识别特定的生物化学过程。
在本研究中,测试案例阻抗谱彼此互不相同(图6)。对于AgCNP1(图6A-B),在8小时消毒期内收集的光谱用于如上所述的感染性测定。频谱示出了近乎一致的阻抗特性,只有在高频(随时间减少;100Hz至100kHz)时幅度上的差异才是明显的,以Bode表示。在相位与对数(频率)表示中,相位峰值中有明显的、时间依赖的、向更高频率的偏移。这些结果被限制在α-分散区:我们预期光谱变化与离子扩散有关,特别是在细胞膜以及与细胞膜的物理相互作用中。类似峰光谱特征代表了具有不同时间常数的两个物理过程的叠加,这可以通过拟合和电路建模(下文)归因于细胞膜上的特定变化。AgCNP2(图6D-E)在4小时的孵育期内示出了相似的初始光谱特征(两个成分)。然而,随着孵育时间的增加,光谱变得更加复杂:表现为两个可观察到的“峰”,它们可以被分解成一个四-成分函数。这些光谱之间的差异证实了不同的颗粒-病毒相互作用,并表明存在另外的物理元件。考虑到观察到的向更高频率的相移,数据表明存在恒定的相位元件成分(在增加频率时,由电阻支配阻抗)。光谱的拟合用所有测试案例的通用图结构证明了这些特征,除了在独特的时间点(图6C,图6F)上特定的AgCNP和病毒相互作用的可变元件(在图6C,图6F中用虚线表示)。可变元件适合作为AgCNP1:OC43的并联电阻和电容,作为AgCNP2:RV14的恒定相位元件,如光谱的相位对阻抗特性所示。特别的,并联元件符合AgCNP1:OC43相互作用的时间依赖性行为,其值从高电阻和中等电容变化到显著较低的值。特别是,电阻值随着孵育而急剧变化。结果共同证实了提出的颗粒:病毒相互作用导致膜完整性/渗透性的变化;降低的电阻与降低的膜密度的渗透性和电容,以及与氧化物纳米颗粒的物理相互作用有关。RV14:AgCNP2的恒定相位元件可变成分是一个频率依赖性的模拟不完美的电介质的元件。在这个系统的案例中,增加孵育组导致模型电介质的特性越来越不完善:导致电阻特性从初始的特性演变为类似于OC43的特性。为了更好地解释且将观察到的原位特征赋予给独特的物理化学过程,制作了物理模型并研究了独特的控制反应。
5.7开发生物电化学阻抗光谱的物理模型:
针对RV14和OC43病毒系统制作了模拟系统,以确定在原位EIS测量期间产生的独特的抗病毒机制。具体来说,我们希望在病毒和电解质之间的界面上再现病毒的特征。因此,制作了两个独特的系统来模仿RV14表面的致密蛋白质结构和OC43的包膜表面。对于与RV14相关的测量,使用牛血清白蛋白,而脂质体用于OC43的脂质膜。所有测量均在与原位测量相同的电解质条件下进行,以控制基于溶液的阻抗贡献(即0.1M Tris-HCl,pH 7.5)。脂质体通常用于病毒研究,包括作为药物/基因递送疗法的病毒模拟载体,作为病毒样颗粒。在当前的研究中,脂质体被合成为OC43冠状病毒的近似尺寸(约120nm),以适当地模拟AgCNP和脂质体之间的任何物理相互作用。在每个试验案例中,将病毒模拟材料分散在溶液中,并以类似于用于原位病毒测量的方案的方式滴涂到玻璃碳电极的表面。在每种情况下,模拟材料的行为似乎反映了观察到的相关病毒的行为,具有相应的AgCNP制剂依赖性。图4示出了收集的在感染性试验中有效的病毒:颗粒对的病毒类似物测量的EIS谱。值得注意的是,拟合的光谱导致类似于原位数据的等效电路。特别是,电路图与原位研究中产生的电路图完全相同,只有图右侧的元件保持可变。对于脂质体/AgCNP1系统(图7A,图7F),我们观察到可变元件是一个并联电阻和电容,并在孵育期间保持这种特性。然而,我们看到这些元件的值在孵育期内发生了变化从而导致相关的相移,这是由于其特性从更具电容性变为电阻性。出现了BSA/AgCNP2系统的相关拟合材料(图7E,图7G)并与RV14/CNP2原位数据相关。然而,我们看到模拟系统中的光谱比在病毒系统中看到的光谱更不清晰。性质上的微小差异可归因于系统之间的小规模(拓扑)差异。特别是,BSA是单个球状蛋白,而RV14是蛋白质的聚集体,具有更粗糙的表面拓扑结构。光谱之间的差异可归因于不同的物理化学环境,然而,光谱表明颗粒和病毒/类似物之间的总相互作用是相似的。考虑到模型中额外的电阻特性的演变,我们决定识别发生的任何特定化学变化。因此,已知通过同时按比例生产超氧化物和过氧化氢来诱导脂质过氧化,氧自由基产生系统被用作活性的正控制(positivecontrol)。
在这些实验中,通过光谱中的相关变化来评估对自由基氧释放的正控制的效果。观察到(图7B)对于BSA/AgCNP2光谱(图7C),氧化再现了在RV14/AgCNP2系统中观察到的额外的峰。观察到的特征也在Lipo/AgCNP1系统中重现(图8),证实了病毒系统中的光谱特征变化不是源于AgCNP1孵育中的化学攻击。
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Claims (49)
1.一种可分配的组合物,包括金属缔合氧化铈纳米颗粒(mCNP)和赋形剂。
2.根据权利要求1所述的可分配的组合物,其中所述金属选自银、金、钌、钒、铜、钛、镍、铂、钛、锡和铁。
3.根据权利要求2所述的可分配的组合物,其中所述金属包括银(AgCNP)。
4.根据权利要求3所述的可分配的组合物,其中所述银的量小于10重量%。
5.根据权利要求1至4中任一项所述的可分配的组合物,其中所述赋形剂选自水、氯仿、二氯甲烷、丙酮、甲基乙基酮、环己烷、乙酸乙酯、乙醚、低级醇、低级二醇、THF、DMSO或DMF。
6.根据权利要求5所述的可分配的组合物,其中所述赋形剂包括水。
7.根据权利要求1所述的可分配的组合物,其中所述mCNP还掺杂有氟。
8.根据权利要求1至4中任一项所述的可分配的组合物,其中所述mCNP的尺寸小于100nm、小于50nm、小于25nm、小于15nm或小于10nm。
9.根据权利要求1至4中任一项所述的可分配的组合物,其中所述mCNP的尺寸为20nm。
10.根据权利要求1至4中任一项所述的可分配的组合物,其中所述mCNP的尺寸为5nm。
11.根据权利要求1至6中任一项所述的可分配的组合物,其中所述mCNP包括占主导的3+表面电荷。
12.根据权利要求1至6中任一项所述的可分配组合物,其中所述mCNP包括占主导的4+表面电荷。
13.根据权利要求1至12中任一项所述的可分配组合物,其中所述AgCNP通过以下方法来生产,所述方法包括:溶解铈前体盐和银前体盐,如硝酸铈和硝酸银,并氧化已溶解的铈前体盐和银前体盐。
14.根据权利要求1至12中任一项所述的可分配组合物,其中所述AgCNP通过以下方法来生产,所述方法包括:溶解铈前体盐和银前体盐,如硝酸铈和硝酸银;通过含有过氧化物的掺杂剂氧化已溶解的铈前体盐和银前体盐;和通过含有氢氧化铵的掺杂剂沉淀纳米颗粒。
15.根据权利要求1至12中任一项所述的可分配组合物,其中所述AgCNP通过以下方法来生产,所述方法包括(i)溶解铈前体盐和银前体盐,如硝酸铈和硝酸银;(ii)通过含有氢氧化铵的掺杂剂氧化和沉淀已溶解的铈前体盐和银前体盐;(iii)在水中洗涤和再悬浮已沉淀的纳米颗粒;(iv)用过氧化氢处理再悬浮的纳米颗粒;和(v)洗涤来自步骤(iv)的纳米颗粒以除去离子化的银。
16.一种消毒剂制剂,包括权利要求1至15中任一项所述的可分配的组合物、干燥剂、有机酸、表面活性剂、聚合物粘合剂和水。
17.根据权利要求16所述的制剂,其中所述制剂包含一种或多种权利要求1至15中任一项所述的可分配的组合物。
18.根据权利要求17所述的制剂,其中所述制剂包含AgCNP。
19.根据权利要求16所述的制剂,其中权利要求1至15中任一项所述的可分配的组合物的量为约0.1至10重量%。
20.根据权利要求16所述的制剂,其中所述干燥剂选自乙醇和异丙醇。
21.根据权利要求20所述的制剂,其中所述干燥剂为乙醇。
22.根据权利要求21所述的制剂,其中所述乙醇的量为约0至40重量%。
23.根据权利要求16所述的制剂,其中所述有机酸为柠檬酸。
24.根据权利要求23所述的制剂,其中所述柠檬酸的量为约0.5至2重量%。
25.根据权利要求16所述的制剂,其中所述表面活性剂选自月桂基胺氧化物、肉豆蔻胺氧化物、其他两性离子剂、tergitol 15-S-15或其他仲醇乙氧基化物。
26.根据权利要求16所述的制剂,其中所述表面活性剂的量为约0.5至3重量%。
27.根据权利要求25所述的制剂,其中所述表面活性剂为月桂基胺氧化物和tergitol15-S-15。
28.根据权利要求27所述的制剂,其中所述月桂基胺氧化物的量为约0.25至2重量%。
29.根据权利要求27所述的制剂,其中所述tergitol 15-S-15的量为约0至1重量%。
30.根据权利要求16所述的制剂,其中所述聚合物粘合剂选自聚(2-乙基-2-噁唑啉)和聚乙烯吡咯烷酮-乙酸乙烯酯共聚物。
31.根据权利要求30所述的制剂,其中所述聚合物粘合剂为聚(2-乙基-2-噁唑啉)。
32.根据权利要求31所述的制剂,其中所述聚(2-乙基-2-噁唑啉)的量为约1至25重量%。
33.根据权利要求30所述的制剂,其中所述聚乙烯吡咯烷酮-乙酸乙烯酯共聚物的量为约1至30重量%。
34.根据权利要求16所述的制剂,其中所述水的量为约15至45重量%。
35.一种装置,包括将权利要求16至34中任一项所述的制剂置于其中的容器和用于分配可分配的组合物的喷嘴。
36.根据权利要求35所述的装置,其中所述装置还包括与所述可分配的组合物连通的泵。
37.根据权利要求35所述的装置,其中所述容器是流体密封的,并且其中所述可分配的组合物还包括推进剂。
38.一种消毒薄膜,包括放置于基底上的权利要求1至15中任一项所述的组合物。
39.根据权利要求38所述的消毒薄膜,其中所述消毒薄膜能够进行残留的消毒。
40.根据权利要求39所述的消毒薄膜,其中所述残留的消毒发生在15至30分钟内。
41.根据权利要求38所述的消毒薄膜,其中所述消毒薄膜能够进行快速的消毒。
42.根据权利要求41所述的消毒薄膜,其中所述快速的消毒发生在少于1分钟内。
43.根据权利要求36所述的消毒薄膜,其中所述消毒薄膜在1至14天内有效。
44.根据权利要求38至43中任一项所述的消毒薄膜,其中所述消毒薄膜暴露于水时被激活。
45.根据权利要求38至4中任一项所述的消毒薄膜,其中干燥的消毒薄膜可以通过暴露于水而被再激活。
46.一种消毒表面的方法,所述方法包括将权利要求1至15中任一项所述的可分配的组合物分配到表面上。
47.一种织物,包括放置于所述织物表面上的权利要求1至15中任一项所述的组合物。
48.根据权利要求47所述的织物,其中所述织物被配置为包括服装、口罩、手套或任何其它PPE中的至少一部分。
49.一种可分配的组合物,包括CNP和/或AgCNP和赋形剂。
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- 2021-04-30 US US17/921,056 patent/US20230157299A1/en active Pending
- 2021-04-30 CN CN202180043058.2A patent/CN115996633A/zh active Pending
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CA3181177A1 (en) | 2021-10-04 |
WO2021222779A1 (en) | 2021-11-04 |
US20230157299A1 (en) | 2023-05-25 |
EP4125364A1 (en) | 2023-02-08 |
BR112022021510A2 (pt) | 2023-01-24 |
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