CN1070704C - 钙改性氧化纤维素止血剂 - Google Patents
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Abstract
一种钙改性的氧化纤维素止血剂可提供比未改性的或钠或钾改性氧化纤维素更快的止血作用。在改性的氧化纤维素中钙的重量百分数必须在约0.5-4之间范围,以便增强止血作用,同时不太妨碍生物吸收性。在第二个实施方案中,用钙和钠或钾改性氧化纤维素止血剂。第二个实施方案可用于酸敏感材料。
Description
本发明涉及通过用钙或钙和钠或钾的混合物处理氧化纤维素制备可生物吸收的止血剂。
氧化纤维素织物是医用中长期使用的可生物吸收和吸收性织物。自然存在和再生的纤维素都可被氧化。为方便和简短起见,我们将两者都称为″氧化的纤维素″。这种织物的三个例子是Interceed*(TC7)可吸收粘连屏障、Surgicel*可吸收止血剂和Surgicel Nu-Knit*可吸收止血剂(都购于Johnson和JohnsonMedical,Inc.,Arlington,TX)。而且,所有这些材料都是酸性的。1g Interceed屏障悬浮在100ml纯水中的水相的pH值约为4.1。一块用水完全饱和织物表面pH值为约1.7。象凝血酶、组织血纤维蛋白溶酶原活性剂类似物(t-PAA)之类的材料和其它对酸高敏感材料在这样的基质中立即失去活性,妨碍将这些材料用于手术部位。作为选择,如果将这样一种材料由氧化纤维素织物供应,织物必须首先中和。所以,过去为改进其性质改变氧化纤维素的努力一般集中在中和材料上。
Doub等人在美国专利No.2517772中叙述了用凝血酶浸渍中和的氧化纤维素产物。他们公开了一种用碳酸氢钠水溶液(例2)或乙酸钙水溶液(例1)中和氧化纤维素织物的方法。Doub等人专利的例2公开了用强碱性碳酸氢钠水溶液中和氧化纤维素织物,然后用凝血酶浸渍中和的产物。然后将凝血酶浸渍的织物冻结,并由冻结状态干燥,以得到一种高止血手术敷料。该专利没有公开任何有关动物试验的实际数据。
Dol′berg等人,Farm zh.(Kiev)1971,26(2),53-56,研究了氧化纤维素的止血性质,并将该材料与各种药用化合物相结合。他们报道氧化纤维素的钠和钾盐可用于医疗,钾盐优于酸形式之处在于它是中性的并能与许多药用物质相容。
Barinka等人在英国专利说明书1593513(1981,7,15公开)中公开了用硝酸和稳定的(″钝化了的″)硝酸钠混合物氧化纤维素的方法。然后用尿素或其N,N双取代烷基或酰基衍生物的醇-水溶液稳定该材料。最终,氧化纤维素通过用钙、钠或铵的盐酸盐和乙酸盐的等克分子浓度的混和液重复吸收及离心甩干相交替方式转变成其钙、钠或铵盐。
Saferstein等人在美国专利5134229中公开了一种通过用弱酸的弱碱性盐的水醇溶液接触酸性氧化纤维素以提高纤维素材料的pH值到5和8之间来制备可稳定贮藏的氧化纤维素。其弱酸盐优选乙酸钠、乙酸钾、柠檬酸钠、甲酸钠、柠檬钾、甲酸钾、磷酸氢二钠、磷酸氢二钾或其混合物。重复Doub等人的方法,他们发现用碳酸氢钠水溶液中和氧化纤维素织物导致部分胶化的织物改变其原始尺寸,很弱并稍微改变其完整性,织物的抗拉强度太低不能实际应用,例如止血应用。按照Doub等人专利例1的指导用乙酸钙完全中和氧化纤维素织物,他们发现,得到的织物有满意的完整性,但对哺乳动物皮肤和其它体细胞在接触处有刺激性。此外,在织物使用位置形成大的稍白的认为是肉芽肿组织块。显然,这种组织试图封闭氧化纤维素织物的钙盐。Saferstien等人没有公开钙盐作为中和氧化纤维素的选择。
钙在止血中的作用是熟知的;事实上,血凝因子命名法国际委员会已经将钙称为13个血凝因子之一(因子Ⅳ)。钙在原凝血酶转变成凝血酶,血凝结过程的一个步骤中起重要作用。止血方法的详细说明见标准医学教科书,例如Remington′s PharmaceuticalSciences,ed.by A.R.Gennaro(Mack,Easton,PA,1990)。钙实际应用的另外的例子是加强止血。Superstatp*可吸收的胶原止血剂含有钙离子,以提高其止血效果。另一方面,为防止无意中的血凝结,将柠檬酸加到血收集袋中,以螯合血浆中的钙。
尽管作为止血材料的中和氧化纤维素的价值和钙在止血中起的重要作用被公认,在本发明之前,可吸收性钙改性止血剂的目标没有完成。
按照本发明,制备可生物吸收的手术止血剂的方法包括步骤:
(a)使含预定浓度羧酸的氧化纤维材料与弱酸钙盐的无氯化物溶液接触,以中和约5.5-45%的羧酸,和
(b)冲洗该材料以除去多余的盐。
在说明书和所附权利要求书中,除另有规定,百分数均为重量百分数。
由本发明方法得到的止血剂是氧化纤维素织物,它含0.5-4%,最好含约2.0-3.5%的钙。该织物比非改性氧化纤维素或用钾或钠盐中和的氧化纤维素证实有更大的止血作用。钙浓度低于3.5%的止血剂并不象含高浓度织物那样对皮肤和其他体细胞有刺激。
在本发明的另一个实施方案中,制备可生物吸收手术止血剂的方法包括步骤:
(a)使含预定浓度羧酸的氧化纤维素材料与弱酸的钙盐和弱酸的钠或钾盐的无氯化物溶液接触,以中和约50-80%的羧酸,选择盐浓度使钙中和约5.5-45%的酸,而钠或钾盐中和剩余部分,和
(b)冲洗该材料以除去多余的盐。
本发明提供一种氧化纤维素手术止血剂,它比现有技术的酸性或中和的氧化纤维素显示出更大的止血作用。由于使用比现有技术所用材料显著少的钙,所以本发明方法得到的材料显示出比用(大量的)钙中和的氧化纤维素更快的生物吸收性和更大的生物相容性。
制备本发明止血剂的方法与美国专利5134229(′229)(这里结合参考)中叙述的方法有点相似。区别包括用于止血材料组合物改性的碱性盐的用量和成分。在一个实施方案中,该材料用钙盐而不用′229中公开的钠和钾盐处理。在另一个实施方案中,通过添加混合的Ca和Na-或K-盐改性该组合物。在两种情况下,所说的盐都是无氯化物的,避免形成盐酸,盐酸能引起氧化纤维素分解成低分子量聚合物,使抗拉强度损失并缩短贮藏寿命。弱酸基本上无此损害。
钙盐最好是水和醇可溶的乙酸钙,但其它盐也是合适的,例如柠檬酸盐、甲酸盐、丁酸盐和丙酸盐。
该材料改性后,冲洗去除多余的盐。为冲洗,水,醇和其它现有技术中熟知的有机溶剂,及其混合物都是合适的。优选有机溶剂,因为它便于随后的材料干燥。水很少选用,因为材料如果不拉长而干燥会趋于收缩。一般,羧酸类含约18-21%(重量)氧化纤维素原材料,而钙附着并中和羧酸基团。钙盐的量决定中和的程度,优选中和约5.5-45%(重量)酸基团的量。钙提高材料的止血性。但是,如果用的钙太多,织物引起刺激。优选的钙浓度为约2.0-3.5%(重量)。
有时需要更完全中和的材料。例如,如果将酸敏感部分加到材料上,那么需要更高的pH值。对于使用凝血酶的织物,例如,pH值应在约5.0-8.0的范围内,优选范围为约6.5-7.0。pH值的测量如下:将1g材料试样置入150ml烧杯中,加入最近煮沸并冷却的纯水,将试样搅拌随后静置5分钟,之后将液体倒入清洁干燥的烧杯中并测量液体的pH值。优选使用钠或钾盐以提供更高的中和作用,而不产生引起刺激的织物。合适的盐包括乙酸钠、柠檬酸钠、甲酸钠、磷酸氢二钠、乙酸钾、柠檬酸钾、甲酸钾、磷酸氢二钾及其混合物。更加完全中和的织物中钠或钾的浓度优选在约2-6%范围内。
制备本发明中和的氧化纤维素的优选方法是首先使酸性氧化纤维素织物与经选择用量和克分子浓度使5.5-45%羧酸基中和的钙盐溶液接触。要制备更完全中和的材料,所用钠和/或钾盐与钙盐一起的用量和克分子浓度优选提供约50-80%的中和作用。虽然可使用水或醇溶液,但优选水和醇的混合液。
当将酸敏感药物加到织物时,一般通过将织物浸渍到药物盐溶液中使织物饱和来完成这种添加。合适的药物包括凝血酶、血纤维蛋白原和抗纤维蛋白溶酶,如抑肽酶。优选添加凝血酶以提供强的止血。1000u/ml凝血酶盐溶液可购得并是适合的。
实施例
下面实施例说明本发明方法和产品。该实施例不能解释为限制本发明的范围,而是有助于完全说明本发明。
实施例1制备含0.5%(重量)钙的钙改性的NU-KNIT*
将50克含20%羧酸(0.22摩尔羧酸)的Surgicel Nu-Knit*织物切成30.5cm宽,缠绕在塑料的多孔芯上并放入容器中。将1600ml异丙醇(IPA)(1257.6克)和1600ml蒸馏水(1600克)加到Haake型FS循环槽中。该循环槽连接到容器上,以致使溶液流到容器底部和多孔芯上,通过缠绕芯的多层织物渗出。
将1.07克乙酸钙一水合物(0.006摩尔)溶入25ml蒸馏水:25ml的IPA中。立刻都加到循环槽中作为绕织物的循环溶液。一水乙酸钙在水溶液(w/w)中的浓度:IPA为1.07/2902.25×100=0.0368%。溶液循环2小时后停止反应。
将芯从容器中移出并放入带有800ml IPA的筒中。搅拌30分钟,除去溶剂并用新鲜的异丙醇代替,搅拌30分钟。取出织物并在试验室通风柜中空气干燥。通过原子吸收谱分析织物试样的钙,得出具有0.45%(重量)的钙。织物滴定羧酸含量表明该织物羧基含量为18.9%,表明中和度为5.5%。
实施例2制备含约0.8%(重量)钙的钙改性Surgicel*
将48.6克带20%羧酸含量(0.216摩尔羧酸)的Surgicel*切成10cm宽,缠绕在30.5cm长多孔芯上,并放入容器中。该容器连到一个Haake型FS循环槽上。将1850ml的IPA(1454.1克)和1850ml的(1850克)的蒸馏水加到槽中。使溶剂绕织物循环几分钟。
将1.90克一水乙酸钙(0.010摩尔)溶入40ml IPA和40ml蒸馏水中。将该溶液加到滴液漏斗中,并在1/2小时期间慢慢加到Haake循环槽中。然后使溶液绕织物循环2小时。溶液中一水乙酸钙的浓度wt/wt(乙酸钙的重量/溶剂的重量)为1.9/3375.5×100=0.056%。2小时后从容器中移出织物并放入装有100%IPA的刻度筒中,绕着织物搅拌IPA溶液30分钟,除去水;然后用新鲜的IPA代替溶剂并绕着织物再搅拌30分钟。将织物悬挂在通风柜中空气干燥。通过原子吸收测量钙的浓度为0.79%(重量)。织物滴定羧酸含量显示18%羧酸含量或10%的中和度。
实施例3制备含2.3%钙的钙改性Surgicel*
将49.4克含20%羧酸(0.219摩尔羧酸)的Surgicel切成10cm宽,缠绕在塑料芯子上并放入一个容器中。将1850ml蒸馏水(1850克)和1850ml IPA(1454.1克)添加到循环槽中并绕容器和织物循环5分钟。
将5.80克一水乙酸钙(0.033摩尔)溶入100ml异丙醇/100ml蒸馏水中。将其加到滴液漏斗并在30分钟期间慢慢滴入循环溶剂中。溶液中乙酸钙的浓度wt/wt是0.166%(5.8/3482.7×100)。使溶液绕织物循环2小时;然后从容器中取出芯子。
将该芯子放入1升的圆筒中并充入800ml的100%IPA。用一个磁力搅棒绕芯子将溶液搅拌30分钟,从织物中去除水。除去溶剂并用800ml新鲜IPA取代,搅拌30多分钟。然后将织物悬挂在通风柜中空气干燥。分析织物钙为2.3%(重量)钙。滴定显示14.6%羧酸含量或27%中和度。
中和度表示该重量百分数的原羧酸基转变成钙盐。剩余的羧酸基不变,并以游离酸存在。
实施例4-8制备具有不同的钙百分数的钙改性Surgicel*和Surgicel NU-KNIT*。
按照实施例1-3的方法,不同之处是乙酸钙浓度变化,以提供0.8%、2.3%和5.5%钙的钙改性Nu-Knit*和0.5%和5.5%钙的Surgicel*。用猪脾切口模型测量所有8个试样和非改性对比试样的止血结果示于表1。
表1
Nu-Knit*和钙改性Nu-Kint*的止血研究出血时间分 Nu-Kint Nu-Knit Nu-Knit Nu-Knit Ku-Knit
对比 用0.5%钙 用0.8%钙 用2.3%钙 用5.5%钙
9.0 5.0 4.5 3.50 3.25
7.0 4.5 5.0 3.25 3.00
5.0 5.0 4.0 3.50 3.25
6.0 4.5 4.5 4.00 3.50
6.5 5.0 5.0 4.50 3.00
6.5 5.5 5.0 3.50 3.00
平均 6.6 4.9 4.6 3.7 3.1
Surgicel*和钙改性Surgicel*的止血研究
Surgicel Surgicel Surgicel Surgicel Surgicel
对比 用0.5%钙 用0.8%钙 用2.3%钙 用5.5%钙出血时间分
14.0 12.0 11.0 8.25 6.0
11.0 11.5 10.5 9.25 6.5
11.0 12.0 11.5 9.75 5.25
12.5 10.5 10.0 8.0 6.5
13.0 11.0 9.5 8.5 6.0
14.0 10.5 10.0 8.5 7.0平均 12.5 11.25 10.4 8.7 6.2
实施例9 ORC的″过量的″钙改性
重复美国专利No.2517772(Doub等人)的实施例6制造钙中和的氧化纤维素织物。通过原子吸收分析该织物的钙含量,得出含6.75%的钙。
将按照本发明制造的氧化纤维素织物(由原子吸收测得它含2.77%的钙)和按照Doub等人的实施例6制造的织物(含6.75%钙)植入鼠的皮下组织,手术14天后评定吸收性和组织接受性。本发明的钙ORC仅引起通常见于用Surgicel*的很轻度的组织反应。该材料在大多数动物上被完全吸收,仅在几个动物上发现少量软的,胶状的材料。由Doub等人的实施例6制造的钙ORC一贯地引起严重的反应,形式为大的适度硬的、稍白的、非纤维块,认为是内芽组织和囊组织。此外,植入在14天后没有完全吸收。
按Barinka等人(Brit.Pat,Spec.1593513)叙述的实施例研究表明它形成的钙改性ORC的性质相似于Doub等人的。
实施例10钙改性ORC与钠改性ORC比较
测量钙改性的Surgicel*和Nu-Knit*的止血作用与相似地钠改性Surgicel和钠Nu-Knit的相比较。所有织物都相似中和到相应程度。通过用乙酸钙处理改性Surgicel和Nu-Knit织物,中和氧化纤维织物上约30%的羧酸类。这样产生的织物含2.6%的钙。用乙酸钠中和Surgicel和Nu-Knit,产生含2.9%钠的织物,它对应约30%的中和度。
用猪脾切口模型测量止血作用。表2示出了止血试验的结果。结果是5个出血场合的平均。
表 Ⅱ
试验材料 止血时间
Surgicel 8.0分
Nu-Knit 5.0分
钙Surgicel 5.5分
钙Nu-Knit 3.3分
钠Surgicel 6.5分
钠Nu-Knit 4.5分
试验表明钠和钙改性的氧化纤维素织物都比相应的原始材料更快地止血,但是,钙改性的氧化纤维素织物比同等钠中和的织物更快。这可解释为钙在止血中起作用。
实施例11钙/钠混合的氧化纤维素织物
将0.984克乙酸钙水合物和1.52克三水乙酸钠加到含170ml IPA和170ml蒸馏水的树脂釜中。搅拌内容物直到两种盐都溶解。盐各存在以摩尔量中和氧化纤维素上30%羧酸类。将羧酸含量为20%的8.4克Surgicel织物加到该溶液中,并将溶液搅拌4小时。然后移出织物,并用两批1650mlIPA冲洗。将织物在空气中干燥。按前面所述测量在蒸馏水中的织物的pH值为5.58。用原子吸收分析织物的钙和钠为2.03%的钙和2.2%钠。织物的pH比如果仅用钙更高(一个仅用30%化学计量的乙酸钙改性的类似的织物在蒸馏水的pH显示为4.1)。在这个实施例中,该织物的pH足够高,致使它可与酸敏感药物相容,并还具有足够的钙增强止血,没有超出上述3.5%浓度的产生组织刺激的钙。
Claims (11)
1.一种可生物吸收的手术止血剂,它包括含约0.5-4.0%的钙的中和氧化纤维素织物。
2.权利要求1的手术止血剂,其中上述织物含约2.0-3.5%的钙。
3.权利要求1的手术止血剂,进一步含约2-6%的钠或钾。
4.权利要求3的手术止血剂,它进一步含止血有效量的凝血酶。
5.一种制备可生物吸收的手术止血剂的方法,包括步骤:
(a)使含预定浓度羧酸的氧化纤维素材料与预定量的具有预定选择摩尔浓度的弱酸钙盐的无氯化物溶液接触,中和约5.5-45%羧酸并提供织物中钙含量为约0.5-4.0%;和
(b)冲洗上述材料除去多余的盐。
6.一种制备可生物吸收的手术止血剂的方法,包括步骤:
(a)使含预定浓度羧酸的氧化纤维素材料与预定量的弱酸钙盐和弱酸钠或钾盐的无氯化物溶液接触,中和约50-80%羧酸,选择盐浓度应使钙中和约5.5-45%的酸和钠或钾盐中和剩余部分,并提供织物中的钙含量为约0.5-4.0%;和
(b)冲洗上述材料除去多余的盐。
7.权利要求6的方法,包括另外的步骤:用止血有效量的凝血酶浸渍干燥的材料。
8.权利要求3的手术止血剂,其中上述织物含约2-3.5%的钙。
9.权利要求8的手术止血剂,其中上述织物pH值为5.0-8.0。
10.权利要求9的手术止血剂,其中上述织物pH值为6.5-7.0。
11.权利要求10的手术止血剂,它进一步含止血有效量的凝血酶。
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