CN1215305A - 从生物液中去除补骨脂素的方法 - Google Patents

从生物液中去除补骨脂素的方法 Download PDF

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CN1215305A
CN1215305A CN97193674A CN97193674A CN1215305A CN 1215305 A CN1215305 A CN 1215305A CN 97193674 A CN97193674 A CN 97193674A CN 97193674 A CN97193674 A CN 97193674A CN 1215305 A CN1215305 A CN 1215305A
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K·H·李
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Abstract

本发明涉及去除补骨脂素和补骨脂素降解产物的方法。本发明方法用于任何用补骨脂素处理过的生物液,包括血液,血液级分和从中产生的成分。根据本发明方法处理的生物液基本上没有任何残留的补骨脂素或补骨脂素降解产物。

Description

从生物液中去除补骨脂素的方法
发明背景
近来，由于涉及捐赠血液的潜在危险，正在活跃地研究着使可能在供血者血液或者血液成分中发现的病原性试剂灭活的方法。一项最前途的研究是通过光化学处理灭活病原体试剂。在大多数光化学处理方法中的一个主要的问题是将处理过的血液中的残留的光敏剂或者其分解产物减少到足够低的水平，使得处理过的血液或血制品能被转输给患者。即使对所有的供血者血液已经检测了被已知的病原体造成的可能的污染，但是常常不能从供血者血库中完全去除所有的污染的血液。
这是由几个条件引起的。例如当一个人感染了病毒时，例如引起AIDS的人免疫缺陷病毒(HIV)，存在一段时期，在该时期期间，抗-HIV抗体滴度对于用常规筛选试验的阳性检测太低。因此HIV感染者供出的血液在该时期内可以通过抗体筛选检测，并且可能感染这种供出的血液或者从中制备的血制品的所有受者。还总是有未知的或者未检测到的病原体污染供出的血液的可能性。因为这些原因，近来急切需要一种方法，能从用于人类的捐赠的血液或由其得到的血液成分中除去未检测到的病原体的方法。
Wiesehahn等(美国专利4727027；4748120；和5176921)和Isaacs等(美国专利5139940)描述了在补骨脂素衍生物例如8-甲氧基补骨脂素(8-MOP),4’-羟基甲基-4,5’,8-三甲基补骨脂素(HMT)，4’-氨基甲基-4’,5’,8-三甲基补骨脂素(AMT)，或者其它补骨脂素衍生物存在下，通过UVA照射在生物液中灭活病原体的方法。在该方法中，只有加入的补骨脂素化合物的总量的一小部分在灭活那些病原体中被消耗，加入的补骨脂素化合物的剩余部分或者作为源补骨脂素化合物留在处理过的血液中或者作为补骨脂素分解产物而留在处理过的血液中。
处理过的血液或者血液成分中的这些残留化合物的量可能非常大，并且当用这种处理过的血液或血液成分对患者转输时，患者可能暴露给补骨脂素或补骨脂素分解产物。这种对于补骨脂素或补骨脂素降解产物的暴露可以对患者引起不期望的作用，例如光毒性或者其它与补骨脂素和其分解产物相关的毒性作用。因此，非常希望在所有的人使用之前，从处理过的血液或血液成分中去除残留的补骨脂素衍生物或补骨脂素的分解产物。
近来没有公开过证明从血液和血制品中去除补骨脂素化合物和其分解产物的方法。
发明概述
本发明涉及从补骨脂素处理过的生物液，包括但不限于血液和血制品中去除补骨脂素化合物和其分解产物的方法。本发明方法使用一种与补骨脂素处理过的生物液例如血液或血制品接触的补骨脂素-吸附剂材料。含有补骨脂素化合物或其分解产物的生物液、血液或血制品根据本发明方法处理，以产生基本上没有补骨脂素化合物或补骨脂素分解产物的生物液、血液和血液成分。
附图简要说明
图1所示的是该吸附装置使用的总的图示。第一个容器(9)含有已经照射过的血液或血液成分(11)，其含有在早期紫外A照射期间的残留的补骨脂素或补骨脂素衍生物例如8-MOP,AMT,HMT或其它补骨脂素衍生物和其分解产物。处理过的液体(11)通过泵(13)泵入通过吸附装置(1)，其中残留的光敏剂(几种光敏剂)或者其副产物被去除，进入第二个容器(10)。
图2所示的是吸附装置(1)的垂直剖面图。该柱体由进口帽(4)，出口帽(5)，罩体(6)，两个不锈钢筛(7)和吸附剂(8)组成。柱体内不锈钢筛(7)含有树脂小球，并且防止它们从柱体中漏出。
图3所示的是吸附装置(1)的水平剖面图。吸附剂(8)是直径大小大约0.1-2mm的微孔小球，并且其由聚苯乙烯或者聚苯乙烯与二乙烯基苯共聚合制备。这些微孔小球具有的孔大小在分子水平内，10-1000埃，并且大孔表面积是每克吸附剂100-1000平方米。好的例子是Rohm和Haas公司制备的XAD-4和XAD-16树脂小球。
图4所示的是本发明另一个设计的剖面图。这里吸附剂(14)由微孔纤维(14)制成而不是小球。纤维可以是织物结构或者非织物结构。通过用纤维代替小球可以省去不锈钢筛(7)。
图5所示的是图4所示相同装置的剖面图。这里示出了吸附剂纤维的剖面。这些吸附剂纤维是带有其它细线的织物。
发明的详细描述
本发明的目的是给出一种从生物液例如处理过的血液或血液成分中去除如果残留的光敏剂例如补骨脂素或其衍生物和其分解产物的方法，从而使处理过的生物液在基本上没有残留的光敏剂的情况下转输给患者。适合在本发明方法中使用的生物液包括但不限于全血、血清、血浆、血液级分例如血小板，红细胞和血沉棕黄色层，血液或血液级分的提取物，例如从中纯化的蛋白质，和用一种或几种补骨脂素化合物处理过的任何生物液。
多种补骨脂素吸附材料适合在本发明方法中使用，并且可以制备这些材料的不同的物理形式，并且适合在本发明方法中使用。例如，微孔小球或纤维形式的活性炭是好的补骨脂素吸附剂。但是发现活性炭也可以吸附其它来自血液或血制品的成分。因此，其在本发明方法中的应用只有在如果活性炭不是也去除处理过的生物液的期望的成分的情况下才适合。在本发明方法中使用的优选的吸附剂材料是吸附补骨脂素和补骨脂素分解产物并且具有最小的吸附其它期望成分例如用于人的血液和血制品的成分的容量的吸附材料。
微孔聚合物小球例如由聚苯乙烯和聚苯乙烯与二乙烯基苯共聚合制备的那些小球是在本发明方法中使用的用于补骨脂素、补骨脂素衍生物和其光分解产物的优选的吸附材料。
实际上任何液体适用于本发明方法这一点对于本领域技术人员是显而易见的。具体地说，已经用补骨脂素化合物处理过的所有的生物液适用于本发明方法。通常暴露给补骨脂素化合物的生物液包括但不限于全血、血浆、血清和从血液或血液级分分离的任何成分。补骨脂素化合物已经被用于各种各样的目的，包括将人血和由血产生的产品灭菌以防止传播肝炎病毒、疱疹病毒、HIV和由血液供者产生的任何其它感染或致瘤的病种；将细胞培养产生的生物产物灭菌，例如干扰素、酶、激素和疫苗，以灭活所有的病毒或核酸杂质；和通过用补骨脂素治疗患者后在体外循环中照射血液，接着将补骨脂素处理过的血液送还患者的对人的治疗。
各种不同的补骨脂素-吸附剂材料适用于本发明方法这一点对本领域技术人员也是显而易见的。合适类型的补骨脂素-吸附剂材料的例子包括但不限于活性炭小球或纤维，其包被有或者没有包被生物相容材料、离子交换树脂，例如Dowex小球(从Midland Michigan的Dow化学品公司购得)，和离子交换树脂小球(由Philadelphia Pennsylvania的Rohm和Haas公司购得)，最优选聚苯乙烯和聚苯乙烯与二乙烯基苯共聚合产物。
补骨脂素处理过的生物液以各种各样的方式与补骨脂素-吸附剂材料接触这一点对于本领域技术人员是显而易见的。例如，生物液可以以批量的方式与补骨脂素-吸附剂材料混合，接着通过标准分离方法例如过滤或者重力分离去除补骨脂素-吸附剂材料。另一种可替代的方法是，补骨脂素-吸附剂材料可以放置在标准的色谱装置例如一个柱子中，通过该柱子通过含有补骨脂素的生物液。
实际上所有适用于生物液的补骨脂素化合物适用于本发明方法这一点对于本领域技术人员是显而易见的。补骨脂素化合物是本领域公知的并且描述于美国专利4321919；和美国专利4960408中。通常使用的补骨脂素化合物包括但不限于补骨脂素；8-甲氧基-补骨脂素；4,5’,8-三甲基补骨脂素；5-甲氧基补骨脂素；4-5’二甲基-补骨脂素；4,8-甲氧基补骨脂素；4-甲基补骨脂素；4,4-二甲基补骨脂素；4’-羟基甲基-4,5’,8-三甲基补骨脂素；和4’-氨基甲基-4,5’,8-三甲基补骨脂素。
提供下面的实施例以详细说明本发明，但是本发明不限于此。实施例1
在证明苯乙烯或苯乙烯共聚物小球对于补骨脂素衍生物的吸附容量的该试验中，一个玻璃移液管被用作树脂容器，并且玻璃棉被用来代替不锈钢筛以将小球保留在移液管中。共8克XAD-4树脂小球(从Rohm和Haas公司购得)装入移液管。玻璃棉球放置在移液管内树脂床的底部和顶部。树脂小球的总的床体积是11.4ml。制备几加仑0.5μg/ml AMT(补骨脂素)水溶液，加压通过该小的XAD-4树脂柱，随着时间测定洗脱液中AMT浓度。结果见表1。
                            表1XAD-4树脂柱上AMT吸附
 次数 灌注速度mL/min 最后样品中漏出量的百分数 处理的总体积
    1     19.5     2.4     800
    2     5.7     0.0     1,370
    3     10.2     0.0     1,230
    4     21.1     7.4     1,254
    5     35.5     0.0     1,414
    6     50.1     4.1     980
  总计7,048
用相同的柱体以6个不同的流速进行该试验。由于流速提高，树脂柱体内灌注液的滞留时间减少，使得发生吸附的时间少。因此，如果吸附速度慢或者容量低，则洗脱液中的AMT浓度应该增加。该试验结果表明，洗脱液中的AMT浓度实际上是零，并且不受流速提高的影响。这些结果表明XAD-4树脂小球对AMT在容量和吸附速度两方面具有非常高的亲和性。

Claims (7)

1、从生物液中去除补骨脂素化合物和补骨脂素分解产物的方法，包括：
a)使含有补骨脂素或补骨脂素分解产物的生物液接触补骨脂素-吸附剂材料；和
b)从补骨脂素-吸附剂材料中分离和收集生物液，以提供基本上没有补骨脂素或补骨脂素分解产物的生物液。
2、权利要求1的方法，其中生物液是血清。
3、权利要求1的方法，其中生物液是血浆。
4、权利要求1的方法，其中生物液是血红细胞。
5、权利要求1的方法，其中生物液是全血。
6、权利要求1的方法，其中补骨脂素-吸附剂材料选自聚苯乙烯和聚苯乙烯二乙烯基苯共聚物。
7、权利要求1的方法，其中补骨脂素是8-甲氧基补骨脂素。
CN97193674A 1996-04-09 1997-04-08 从生物液中去除补骨脂素的方法 Pending CN1215305A (zh)

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US6228995B1 (en) 2001-05-08
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