CN1279605A - 脂蛋白氧化的抑制 - Google Patents
脂蛋白氧化的抑制 Download PDFInfo
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- CN1279605A CN1279605A CN98811267A CN98811267A CN1279605A CN 1279605 A CN1279605 A CN 1279605A CN 98811267 A CN98811267 A CN 98811267A CN 98811267 A CN98811267 A CN 98811267A CN 1279605 A CN1279605 A CN 1279605A
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- Prior art keywords
- oxidation
- ldl
- metabolite
- hydroxylation
- lipoprotein
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Abstract
胆固醇降低剂的羟化衍生物抑制脂蛋白氧化,因此对防止动脉粥样化形成和由此产生的血管疾病(包括心脏病发作)的发展有用。
Description
本发明的领域
本发明涉及抑制脂蛋白氧化从而减缓或阻止动脉粥样化形成的方法。该方法需要使用已知胆固醇降低剂的羟化衍生物。
本发明的背景
动脉粥样硬化心血管疾病和相关疾病以及高脂血症相关的疾病事件是残疾和死亡的主要原因。现在已经公认,在健康哺乳动物和已患有高脂血症的个体中降低某些形式的胆固醇,能引人注目地减少心脏病发作、血管疾病和动脉粥样硬化相关的其它疾病。
高脂血症是一种以血清脂质例如胆固醇、甘油三酯和磷脂异常增为特征的疾病。这些脂质不能在血浆中以溶液自由循环,而与蛋白质结合,作为称作脂蛋白的大分子复合物进行运输。脂蛋白按其密度大小分为五类:乳糜微粒,极低密度脂蛋白(VLDL),低密度脂蛋白(LDL),中等密度脂蛋白(IDL),和高密度脂蛋白(HDL)。
高脂血症的一种形式是高胆固醇血症,其特征是存在LDL胆固醇水平升高。高胆固醇血症的初期治疗经常是变更食物为低脂肪和低胆固醇的食物,结合适当的体育运动,之后,当仅通过食物和运动没有达到降低LDL的目标时采用药物治疗。通常认为LDL是“坏”胆固醇,而HDL是“好”胆固醇。尽管人们希望降低升高了的LDL胆固醇水平,但也希望增加HDL胆固醇水平。已经发现,HDL胆固醇水平增加通常和冠心病(CHD)危险降低相关联。
虽然LDL胆固醇被认为是坏的,并且大多数胆固醇降低剂通过降低LDL形式的血浆浓度起作用,但在动脉粥样化形成的早期还有另一个关键过程,那就是LDL的氧化。VLDL和HDL的氧化也会发生,这也促使动脉粥样化形成。氧化会导致细胞内钙增加、能量产生降低、细胞因子激活、膜损伤,所有这些都导致细胞凋亡、坏死和最终细胞死亡。
典型的氧化反应起始于一个活性自由基从LDL颗粒中的多不饱和脂肪酸摄取一个氢原子。由此形成脂质过氧化氢和烷氧基自由基,它们反过来又能启动邻近的脂肪酸氧化,导致脂质过氧化的蔓延。这些氧化形式的脂蛋白比天然脂蛋白更迅速地被巨噬细胞吸收,这样就导致了巨噬细胞胆固醇积累,随后引起泡沫细胞形成和组织巨噬细胞和内皮细胞运动性的抑制。这一系列事件导致血管机能障碍和动脉粥样硬化损伤的形成和活化。
现在我们发现,普遍使用的胆固醇降低剂的某些羟基取代衍生物是脂蛋白的有效抗氧化剂。此外,这些化合物有助于自由基的清除和金属离子的螯合,而自由基和金属离子也是脂蛋白氧化的机制。
本发明概述
本发明提供了一种在哺乳动物中抑制脂蛋白氧化的方法,其包括施用抗氧化有效剂量的羟化胆固醇降低剂。本发明还提供了一种在哺乳动物中清除自由基的方法,其包括施用自由基清除剂量的羟化胆固醇降低剂。本发明还提供了一种抑制脂蛋白螯合金属离子的方法,其包括施用有效剂量的羟化胆固醇降低剂。
在一个优选实施方案中,所述方法通过采用抑制素、特别是atorvastatin的羟化形式而实现,这些化合物在美国专利5,385,929中有描述,所述专利引入本文作为参考。
在另一个实例中,采用羟化氟伐他汀,如有如下结构式的化合物:
附图的简要说明
图1.Atorvastatin及其羟化代谢物的结构式。
图2.吉非贝齐及其羟化代谢物的结构式。
图3.Atorvastatin及其羟化代谢物对铜离子氧化系统(A)、AAPH氧化系统(B)和J-774 A.1巨噬细胞氧化系统(C)中的LDL氧化的影响。在所有三个氧化系统中LDL(100μg蛋白/mL)与递增浓度的所述药物或其代谢物在37℃孵育4小时(系统A和B),或与所述细胞孵育20小时(C)。孵育结束时,用TBARS分析测定LDL氧化。巨噬细胞介导的LDL氧化通过有细胞时得到的值减去无细胞时得到的值来计算。结果以平均值±标准差(SD)(n=3)形式给出。
图4.Atorvastatin及其羟化代谢物对铜离子氧化系统(A)和AAPH氧化系统(B)中的VLDL氧化的影响。VLDL(100μg蛋白/mL)和10μM的atorvastatin或其代谢物在37℃孵育4小时。孵育结束时,用TBARS分析测定VLDL氧化。结果以平均值±SD(n=3)形式给出。
图5.Atorvastatin及其羟化代谢物的自由基清除活性(A)和铜离子螯合能力(B)。A.atorvastatin或其羟化代谢物(20μM)和1mM DPPH孵育并进行517nm吸光度的动力学测定。图中显示了三个有类似型式之不同研究的一个典型实验。采用维生素E(20μM)作为自由基清除剂的阳性对照。B.LDL(100μg蛋白/mL)和atorvastatin或其代谢物(10μM)以及递增浓度的CuSO4在37℃孵育4小时,之后用TBARS分析测定脂蛋白的氧化。结果以平均值±SD(n=3)形式给出。
图6.吉非贝齐和吉非贝齐代谢物的浓度对铜离子氧化系统(A)、AAPH氧化系统(B)和J-774 A.1巨噬细胞氧化系统(C)中的LDL氧化的影响。在所有三个氧化系统中LDL(100μg蛋白/mL)与递增浓度的所述药物或其代谢物在37℃孵育4小时(系统A和B),或与所述细胞孵育20小时(C)。孵育结束时,用TBARS分析测定LDL氧化。巨噬细胞介导的LDL氧化通过有细胞时得到的值减去无细胞时得到的值来计算。结果以平均值±SD(n=3)形式给出。
图7.吉非贝齐及其代谢物对铜离子氧化系统(A)和AAPH氧化系统(B)中VLDL氧化的影响。VLDL(100μg蛋白/mL)和4μM的吉非贝齐或其代谢物在37℃孵育4小时。孵育结束时,用TBARS分析测定VLDL氧化。结果以平均值±SD(n=3)形式给出。
图8.VLDL脂蛋白电泳。在有或无atorvastatin、吉非贝齐或它们的代谢物时铜离子(10μM CuSO4)诱导的脂蛋白氧化之后进行。
图9.吉非贝齐及其代谢物的自由基清除活性(A)和铜离子螯合能力(B)。A.吉非贝齐或其代谢物(20μM)和1mM DPPH孵育并进行517nm吸光度的动力学测定。图中显示了三个有类似型式之不同研究的一个典型实验。采用类似浓度的维生素E(Vit E)作为对照自由基清除剂。B.LDL(100μg蛋白/mL)和吉非贝齐或其代谢物(3μM)以及递增浓度的CuSO4在37℃孵育4小时,之后用TBARS分析测定脂蛋白的氧化。结果以平均值±SD(n=3)形式给出。
图10.吉非贝齐代谢物I和atorvastatin邻羟基代谢物对LDL氧化的联合影响。LDL(100μg蛋白/mL)单独地(对照),或在吉非贝齐代谢物I(3μM)或atorvastatin邻羟基代谢物(4μM)单独或联合存在时,与10μM CuSO4在37℃孵育4小时。之后用TBARS分析测定脂蛋白的氧化。*p<0.01(相对对照)和p<0.01(相对代谢物I),#P<0.01(相对邻羟基代谢物)。结果以平均值±SD(n=3)形式给出。
图11.在富含多不饱和脂肪酸的膜制品中atorvastatin对羟基代谢物的剂量依赖性抗氧化作用。
图12.Atorvastatin对羟基代谢物、维生素E和普罗布考的相对抗氧化能力比较。
图13.Atorvastatin对羟基代谢物和维生素E在膜胆固醇升高的动脉粥样硬化样疾病中的抗氧化能力。
本发明的详细描述
术语“羟化胆固醇降低剂”是指能有效降低哺乳动物LDL胆固醇的任何化学化合物,其在母体结构上至少取代有一个羟基基团,并有抗氧化活性。例如羟化抑制素。所述抑制素是已知的一类HMG-CoA还原酶抑制剂,例如atorvastatin、氟伐他汀和cerivastatin。羟化抑制素是有至少一个羟基取代基团的母体抑制素化合物,例如图1所示的邻羟基atorvastatin和对羟基atorvastatin。其它羟化胆固醇降低剂是羟基取代的fibrates,例如图2所示的羟化吉非贝齐(代谢物I)。用于本发明所述方法的羟化化合物优选有一个羟基连在苯环上的化合物。
高脂血症患者的动脉粥样硬化危险增加起因于其血浆脂蛋白的可氧化性增强。虽然降低血液胆固醇(hypocholesterolemic)的药物治疗,包括3-羟基-3-甲基戊二酰辅酶A(HMG-CoA)还原酶抑制剂(例如atorvastatin)和降低血液甘油三酯(hypotriglyceridemic)的药物苯扎贝特,能降低从高脂血症患者分离的低密度脂蛋白(LDL)对氧化的高敏感性,但在体外用这些药物不能得到这种抗氧化作用。下列实验确定了atorvastatin和吉非贝齐以及特定羟化代谢物对LDL、VLDL和HDL之氧化(即脂质过氧化)敏感性的影响。由铜离子(10μM CuSO4)、自由基产生系统2’2’-偶氮双-2-脒基丙烷盐酸盐(5mM AAPH)或J-774 A.1巨噬细胞样细胞系诱导的脂质过氧化,不被atorvastatin或吉非贝齐母体形式抑制,而被药理学浓度的atorvastatin邻羟基和对羟基代谢物以及吉非贝齐对羟基代谢物(代谢物I)以浓度依赖的方式充分抑制(57%-97%)。通过联合使用atorvastatin邻羟基代谢物和吉非贝齐代谢物I,发现对LDL可氧化性的累加抑制作用。对于VLDL和HDL在以上提到的氧化系统中对氧化的敏感性,上述代谢物得到类似的抑制作用(37%-96%)。这些代谢物对LDL、VLDL和HDL氧化作用的抑制可能与它们的自由基清除活性以及(主要对吉非贝齐代谢物I)它们的金属离子螯合能力相关。此外,HDL氧化的抑制与HDL相关对氧磷酶活性的保持有关。这些数据证明,atorvastatin羟基代谢物和吉非贝齐代谢物I拥有有效的抗氧化潜力,从而保护LDL、VLDL和HDL不被氧化。因此所述羟化胆固醇降低剂通过其抗氧化特性有助于减少脂蛋白导致动脉粥样化的可能性。
在早期动脉粥样化形成中,LDL氧化是一个关键因素,因此,抑制LDL氧化就是抗动脉粥样硬化形成。在氧化应激下也会发生VLDL和HDL氧化,这也促使动脉粥样硬化形成。抗氧化剂既通过遗传也通过环境获得。例如,与脂蛋白相关的食物抗氧化剂,如维生素E、类胡萝卜素或多酚黄酮类化合物,能防止这些脂蛋白氧化。此外,遗传因素,如HDL相关的对氧磷酶,也能保护该脂蛋白不受氧化应激的损伤。降低血液胆固醇的治疗可显著降低高胆固醇血症患者LDL对氧化的高敏感性。因此,降低血液胆固醇治疗之所以被认为有益,可能不仅因为它影响血浆VLDL、LDL和HDL水平,而且因为它能减少导致动脉粥样硬化的氧化脂蛋白的形成。
给高胆固醇血症患者施用HMG-CoA还原酶抑制剂洛伐他汀、辛伐他汀、普伐他汀或氟伐他汀之后,显示了LDL氧化的离体抑制。这些药物对LDL可氧化性的抑制作用被认为起因于血浆“老”的LDL清除增强,“老”的LDL比新合成的LDL更倾向于氧化。这种作用对于抑制素诱导的肝细胞LDL受体活性的刺激和肝VLDL和LDL产生的抑制来说是第二位的。从机理上讲可能还涉及到药物治疗过程中肝产生的母体抑制素代谢物。肝P450药物代谢系统活动通常通过羟基化来参与改变母体抑制素结构。的确,所有上述抑制素除氟伐他汀外,当在与所处理高胆固醇血症患者中观察到的血液药物水平相当的浓度进行测试时,并没有显示对体外LDL氧化的直接抗氧化效果。Atorvastatin,一种新的HMG-CoA还原酶抑制剂,是降低血浆总胆固醇水平和LDL胆固醇水平最有效的抑制素。这种化合物还有降低所有脂蛋白组分的甘油三酯的特性。Atorvastatin治疗增加LDL受体活性并抑制含有载脂蛋白B-100的脂蛋白的直接产生。母体药物及其代谢物都有相对较长的14到36小时的循环半衰期。Fibrate药物也可影响脂蛋白对氧化的敏感性,例如苯扎贝特就有这种能力。所述fibric acid衍生物是脂质调节药物,其能促进富含甘油三酯的脂蛋白的代谢,其次是激活脂蛋白脂酶活性和减少apoC-Ⅲ合成。另一种fibrate,吉非贝齐,已显示不仅能降低血浆甘油三酯,而且能增加人的血浆HDL浓度,降低雄性猕猴的血浆脂蛋白(a)水平。人的吉非贝齐代谢成吉非贝齐酰基葡糖苷酸,在处理后的志愿者血浆和尿液中发现这些代谢物。在吉非贝齐处理的啮齿类动物血浆中发现的吉非贝齐对羟基代谢物(代谢物I)水平比处理的人的相应值高得多,可能反映了剂量和代谢的差异。现在我们显示了atorvastatin和吉非贝齐以及特定羟化代谢物(单独和联合)对LDL、VLDL和HDL氧化敏感性的影响。结果清楚地说明所述药物代谢物(而不是母体药物)对血浆脂蛋白氧化的单独的抑制作用和联合使用时累加的抑制作用。这些数据证明所述羟化衍生物有助于防止脂蛋白氧化和因而减轻其导致动脉粥样硬化的可能性。
通过下列详细实施例说明各种羟化胆固醇降低剂的抗氧化剂活性。
实施例1
材料Atorvastatin及其邻羟基和对羟基代谢物(图1),以及吉非贝齐及其代谢物I(图2)用现有技术方法合成。2.2.-偶氮双-2-脒基丙烷盐酸盐(AAPH)购自Wako化学工业有限公司(大阪,日本)。1,1-二苯基-2-苦基-偕腙肼(DPPH)购自Sigma(圣路易斯,密苏里州)。
脂蛋白 血清VLDL、LDL和HDL从禁食的正常血脂志愿者分离。脂蛋白用不连续密度梯度超速离心制备。所述脂蛋白在其适当的密度(分别为1.006g/mL,1.063g/mL,和1.210g/mL)洗脱,4℃相对150mM NaCl(pH7.4)透析。然后所述脂蛋白通过过滤除茵(0.45μM),于4℃避光保存在氮气中,在2周内使用。在氧化研究之前,所述脂蛋白于4℃在氮气中对无EDTA的PBS溶液(pH 7.4)透析。当用鲎变形细胞裂解液分析(LimulusAmebocyte Lysate assay)(Associated of Cape Cod公司;Woods Hole,麻省,美国)时发现所述脂蛋白没有脂多糖(LPS)污染。脂蛋白的蛋白含量用标准方法测定。
脂蛋白氧化,脂蛋白(100μg蛋白/mL)和10μM CuSO4或5mM AAPH在37℃孵育4小时。AAPH是一种水溶性的偶氮化合物,能受热分解并以恒定速率产生水溶性过氧化氢自由基。加入10μM丁基化对甲酚(BHT)在4℃冷却终止氧化。脂蛋白氧化程度用硫巴比土酸活性物质(TBARS)分析测定,用丙二醛(MDA)作标准曲线。此外,脂蛋白氧化还用脂质过氧化试验来测定,脂质过氧化试验根据脂质过氧化物能把碘化物转化成能用分光光度法在365nm测定的碘来分析脂蛋白的过氧化。LDL氧化动力学通过测定共轭二烯的形成即234nm吸光度增加来连续监测。
巨噬细胞介导的LDL氧化-J-774 A.1鼠巨噬细胞样细胞系从美国典型培养物保藏中心(Rockville,马里兰州)购买。所述巨噬细胞在补充5%热失活胎牛血清(FCS)的达尔伯克氏改良伊格尔培养基(DMEM)中生长。用于脂蛋白氧化研究时,在2μM CuSO4存在下将细胞(1×106/35mm培养皿)与LDL(100μg蛋白/mL)在RPMI培养基(不含酚红)中于培养箱37℃孵育20小时。对照LDL也在相同条件下于无细胞系统中孵育。孵育期结束时,LDL氧化程度在培养基(1000×g离心10分钟之后)中用TBARS分析测定。细胞介导的LDL氧化通过有细胞时得到的值减去无细胞系统得到的值来计算。
脂蛋白电泳,脂蛋白(100μg蛋白/mL)在所述药物存在或不存在下孵育,之后是10μM CuSO4存在下的氧化。然后,用Hydragel-Lipo试剂盒(Sebia,法国)在1%琼脂糖上进行脂蛋白电泳。
自由基清除活性,所述药物的自由基清除能力用1,1-二苯基-2-苦基-偕腙肼(DPPH)实验来分析。每种药物(20μM)和3毫升0.1nmol DPPH/l(溶于乙醇中)混合。然后动力学监测517nm光密度变化的时间过程。
对氧磷酶活性测定,对氧磷的水解速率通过测定25℃时412nm对硝基酚的形成来确定。基础分析混合物包括溶于50mM甘氨酸/NaOH(pH10.5)溶液的1.0mM对氧磷和1.0mM CaCl2。一单位的对氧磷酶活性每分钟产生1nmol对硝基酚。
统计分析-比较两个平均数采用Student t-检验,而比较两组以上数据用方差分析(ANOVA)。数据以平均值±标准差(SD)形式给出。结果
在几个氧化系统中研究了atorvastatin及其羟基代谢物和吉非贝齐及其代谢物对脂蛋白氧化敏感性的影响,所述氧化系统包括含有金属离子(10μM CuSO4)的、能产生自由基的(5mM AAPH)和模拟生物学氧化(J-774 A.1巨噬细胞样细胞系)的氧化系统。
Atorvastatin和脂蛋白氧化-在所有研究的氧化系统中LDL氧化被邻羟基和对羟基atorvastatin代谢物抑制,但不被atorvastatin抑制。这些抑制作用是浓度依赖性的(图3)。在10μM时,邻羟基和对羟基atorvastatin代谢物均抑制LDL氧化,用TBARS分析测定,在CuSO4系统中分别抑制73%和60%(图3A);在AAPH系统中,分别抑制44%和34%(图3B);在巨噬细胞系统中,分别抑制50%和46%(图3C)。在所有研究的浓度和所有氧化系统中,所述邻羟基代谢物是一种比对羟基代谢物更好的LDL氧化抑制剂(图3)。与自由基产生系统(图3B)相比,在金属离子氧化系统(图3A)中两种atorvastatin代谢物均得到更有效的抑制效果。当用脂蛋白相关的过氧化物分析来测定LDL氧化时,在其它氧化系统中得到类似的结果。所述atorvastatin邻羟基和对羟基代谢物降低了LDL相关的过氧化物含量,在CuSO4系统中所述含量从对照LDL的710±51nmol/mg LDL蛋白分别降至192±15和284±13nmol/mg LDL蛋白;在AAPH系统中,所述含量从对照LDL的990±89nmol/mg LDL蛋白分别降至554±32和624±38nmol/mg LDL蛋白。而且,在铜离子(10μM CuSO4)诱导的LDL氧化过程中,234nm处共轭二烯形成的动力学分析揭示,对于对照或atorvastatin处理的LDL,LDL氧化起始所需要的延迟时间是50±7分钟(n=3),而对于两种atorvastatin的代谢物而言,LDL共轭二烯形成在180±25分钟(n=3)后才开始。
图4给出了atorvastatin及其代谢物对VLDL氧化的影响。在铜离子氧化系统中,邻羟基和对羟基代谢物(10μM)分别抑制脂蛋白氧化79%和37%(图4A),而atorvastatin本身则没有影响。在AAPH氧化系统中,这些代谢物的抑制作用仅分别为43%和16%(图4B),atorvastatin本身也没有影响。当用过氧化物形成分析VLDL氧化时,发现类似的结果。所述atorvastatin邻羟基和对羟基代谢物降低了VLDL相关的过氧化物含量,在CuSO4系统中从对照VLDL的1818±333nmol/mg VLDL蛋白分别降至242±22和1088±310nmol/mg VLDL蛋白;在AAPH系统中,从对照VLDL的2169±329nmol/mg VLDL蛋白分别降至1228±210和1819±228nmol/mg VLDL蛋白。类似地,在相同孵育条件下有CuSO4存在的HDL氧化揭示,邻羟基代谢物完全抑制了HDL氧化,而对羟基代谢物抑制大约50%的脂蛋白氧化(表1)。这些代谢物对HDL氧化的抑制作用和对对氧磷酶的保护分别有54%和27%的相关性。与没有加入母体药物时氧化的HDL中的对氧磷酶活性相比,所述HDL相关的对氧磷酶活性显著提高(表1)。
表1.Atorvastatin及其代谢物对HDL氧化和HDL相关的对氧磷酶活性的影响
CuSO4诱导的HDL氧化 对氧磷酶比活
(nmol/mg HDL蛋白) (nmol/mg HDL蛋白/分钟)
MDA 过氧化物对照 9.1±0.1 122±14 26±2Atorvastatin 9.6±0.5 122±15 29±4邻羟基代谢物 0.2±0.1* 9±1* 40±4*对羟基代谢物 4.5±0.3* 65±9* 33±3**p<0.01(相对对照)
所述atorvastatin代谢物对脂蛋白氧化的抑制作用还与自由基清除活性及金属离子螯合能力相关。在DPPH分析中,观察到两种atorvastatin代谢物(20μM)都引起517nm吸光度呈时间依赖性减少,而atorvastatin则没有(图5A)。孵育300秒后,所述邻羟基和对羟基代谢物分别减少517nm吸光度37%和28%。作为比较,20μM的自由基清除剂抗氧化剂维生素E减少517nm吸光度95%(图5A)。这些结果证明atorvastatin代谢物拥有实质的自由基清除能力。
将LDL和递增浓度的CuSO4在37℃孵育2小时以确定过量浓度的铜离子是否能制止atorvastatin代谢物对LDL氧化的抑制作用,以此检验所述代谢物通过螯合铜离子充当LDL氧化抑制剂的能力(图5B)。与对照LDL相比,在所述代谢物存在时在所述孵育系统中加入递增浓度的铜离子仅引起LDL氧化的较小增加(图5B),表明这些代谢物通过螯合金属离子抑制LDL氧化的能力极小。
实施例2
吉非贝齐和脂蛋白氧化
用上述实验测定吉非贝齐及其一种代谢物(代谢物I)对LDL氧化的影响,所述实验与对atorvastatin所作实验类似(图3-5)。在所有研究的氧化系统中LDL氧化被代谢物I抑制,而不被吉非贝齐本身所抑制。代谢物I的这种抑制作用是浓度依赖性的(图6)。在4μM的低浓度时,吉非贝齐代谢物I抑制LDL氧化,在CuSO4氧化系统中为96%(图6A),在AAPH氧化系统中为26%(图6B),在J-774A.1巨噬细胞介导的氧化系统中为99%(图6C),所述LDL氧化用TBARS分析测定。当用所形成的过氧化物数量来分析LDL氧化时,发现类似的结果。吉非贝齐代谢物I减少了LDL相关的过氧化物,在CuSO4系统中从710±57nmol/mg LDL蛋白减至28±7nmol/mg LDL蛋白;在AAPH系统中,从917±78nmol/mg LDL蛋白减至703±38nmol/mg LDL蛋白。而且,对于单独的LDL或有吉非贝齐存在的LDL,LDL氧化起始所需要的延迟时间(用共轭二烯形成的动力学分析测定)是60±9分钟。而与吉非贝齐代谢物I孵育了甚至240分钟之后,在LDL中也没有观察到共轭二烯形成。
分析吉非贝齐及其代谢物对VLDL氧化的影响再次表明了代谢物I(4μM)非常有效的抑制作用,而吉非贝齐没有。所述代谢物I在CuSO4氧化系统中对VLDL氧化抑制96%(图7A),在AAPH氧化系统中抑制91%(图7B)。
在atorvastatin及其代谢物或吉非贝齐及其代谢物存在时的VLDL氧化之后,VLDL的脂蛋白电泳清楚地显示了atorvastatin邻羟基代谢物和吉非贝齐代谢物I降低脂蛋白电泳迁移率的能力(图8)。用LDL和HDL得到了类似的结果。
关于10μM CuSO4存在时HDL氧化,吉非贝齐代谢物I充分抑制了脂蛋白氧化(表2),同时保护对氧磷酶活性,使HDL相关的对氧磷酶活性保持在初始水平(表2)。吉非贝齐本身没有作用。
在无所述药物(对照)或有10μM所述药物存在时,脂蛋白氧化在10μM CuSO4中37℃进行4小时。在与铜离子孵育之前HDL的对氧磷酶活性为50±3nmol/mg HDL蛋白/分钟。结果以平均值±SD(n=3)形式给出。
表2.吉非贝齐及其代谢物对HDL氧化和HDL相关对氧磷酶活性的影响
CuSO4诱导的HDL氧化 对氧磷酶比活
(nmol/mg HDL蛋白)(nmol/mg HDL蛋白/分钟)
MDA 过氧化物对照 9.1±0.1 122±14 26±3吉非贝齐 8.2±0.4 134±13 27±5代谢物I 0.8±0.1* 18±4* 50±7**p<0.0l(相对对照)
为了分析吉非贝齐代谢物I抑制脂蛋白氧化的机制,图中显示了这种代谢物的自由基清除能力(图9A)和铜离子螯合能力(图9B)。通过采用DPPH分析,仅代谢物I,而不是吉非贝齐本身(20μM),显示了517nm吸光度的时间依赖性减少,孵育300秒后光密度减少达86%(图9A)。在吉非贝齐代谢物I存在时LDL和递增浓度的CuSO4在37℃孵育2小时显示,用20μM CuSO4时代谢物I的抑制作用被完全阻止(图9B),表明在该LDL氧化系统中代谢物I对铜离子的螯合在抑制脂蛋白氧化中起作用。
实施例3
Atorvastatin和吉非贝齐之间的相互作用
重复上述一般程序以确定体外加入联合的有效代谢物(吉非贝齐代谢物I和atorvastatin邻羟基代谢物)是否比两种试剂单独使用产生对LDL氧化更大的抑制作用。与对照LDL相比,使用低浓度的吉非贝齐代谢物I(3μM)或atorvastatin邻羟基代谢物(4μM)时,观察到每一种药物对铜离子诱导的LDL氧化仅分别有40%或43%的抑制作用(图10)。然而,当联合使用上述浓度的这些药物时,观察到对LDL氧化88%的显著累加的抑制作用(图10)。
实施例4
在富含多不饱和脂肪酸的膜小泡中评价了atorvastatin对羟基代谢物以及已知抗氧化剂维生素E和普罗布考。取富含二亚麻酰磷脂酰胆碱(DLPC)(浓度为1.0mg DLPC/mL)的500μl膜小泡用于所述脂质过氧化实验。富含DLPC的小泡在HEPES缓冲液(N-(2-羟乙基)哌嗪-N’-(2-乙磺酸))(0.5 mM HEPES,154.0mM NaCl,pH 7.3)中新鲜制备。所述缓冲液不加额外的抗氧化剂(作为对照),或加:(1)各种浓度atorvastatin对羟基代谢物;(2)维生素E;和(3)普罗布考,即4,4’-(异丙叉二硫)-双(2,6-二叔丁基酚)。所述膜小泡溶液立即置于在37℃振荡水浴中。在孵育期间(0-72小时),取100μl样品,加入5.0mM乙二胺四乙酸(EDTA)25μl和35.0mM丁基化对甲酚20μl终止过氧化反应。用一称为CHOD-iodide(Merck,Darmstadt,FRG,Merck Cat.No.14106)的显色试剂通过分光光度法分析测量血清脂蛋白的脂质过氧化物,以确定每个样品的脂质过氧化程度。所述显色试剂组成如下:磷酸钾,pH6.2 0.2M碘化钾, 0.12M叠氮化钠 0.15μM聚乙二醇单[对-(1,1’,3,3’-四甲基丁基]苯基]醚 2g/L烷基苄基二甲基氯化铵 0.1g/L钼酸铵 10μM
给每一个取出的膜小泡样品加入1.0mL CHOD显色试剂,将所述样品避光孵育4小时。测定该溶液365nm的吸光度(ε=2.4×104M-1cm-1)。测量三次脂质过氧化物的形成,数值表示为平均值±SD。不同实验条件下结果之间的差异显著性用双侧Student t检验分析。
图11显示了不同剂量浓度的atorvastatin对羟基代谢物的抗氧化活性。结果说明,所述对羟基化合物有剂量依赖性抗氧化活性,在10.0μM时引起脂质过氧化80%的抑制。甚至在浓度低至10.0μM时,所述对羟基化合物也抑制了高水平(>102μM)的脂质过氧化。
图12所显示的结果证明,atorvastatin对羟基代谢物比其它已知的抗氧化剂,特别是维生素E和普罗布考,显著地更为有效。
Atorvastatin对羟基代谢物的抗氧化活性在膜胆固醇升高的动脉粥样硬化样疾病中增加,见图13。
前述这些实验证明,HMG-CoA还原酶抑制剂(例如atorvastatin)的代谢物和fibric acid衍生物(例如吉非贝齐)的代谢物显著抑制几个氧化系统的脂蛋白氧化。在动脉粥样硬化形成中,LDL氧化是一个关键事件,因为它促使巨噬细胞胆固醇积累和泡沫细胞形成,也促成细胞毒性、血栓形成和炎症。因此抑制LDL氧化有助于减弱动脉粥样硬化过程。尽管没有广泛研究,VLDL和HDL氧化在氧化应激下也会发生,并促进动脉粥样硬化发展。在VLDL,脂质过氧化主要涉及核心甘油三酯多不饱和脂肪酸的氧化,而在HDL,表面磷脂脂肪酸是易于氧化的主要底物。
在高胆固醇血症患者和高甘油三酯血症患者中,高浓度血胆固醇和甘油三酯是动脉粥样硬化的危险因素。危险的增加是因为所述脂蛋白对氧化敏感性增强。有几种降低血液脂质的药物显示能减轻高胆固醇血症患者的高LDL氧化倾向。这种对LDL氧化的抑制作用可能起因于对更易于氧化的“老LDL”的清除增强(通过药物诱导的LDL受体活性增强来达到,主要是在肝中)。此外,这种抗氧化的保护作用可来自体内形成的有抗氧化特性的药物代谢物。但是,当在药理学浓度作体外实验时,除氟伐他汀外,所研究的降低血液脂质的药物中没有一种母体形式的药物展示对LDL氧化的直接抑制作用。以上数据说明,所述母体药物atorvastatin和吉非贝齐在体外不影响LDL、VLDL或HDL的可氧化性,即使在使用高浓度时也是如此。然而,在金属离子依赖性和非依赖性系统中,低药理学浓度的特定羟化代谢物均可诱导对LDL、VLDL和HDL氧化非常有效的抑制作用。与AAPH系统相比,在CuSO4系统中所述药物代谢物对脂蛋白氧化性的抑制作用更为显著,这种现象可能与所述代谢物影响自由基清除和铜离子结合有关。所述吉非贝齐代谢物I和atorvastatin羟化代谢物均表明是有效的自由基清除剂。
与atorvastatin邻羟基代谢物相比,在CuSO4系统中吉非贝齐代谢物I是一种更好的金属离子螯合剂。铜离子浓度增加完全阻止了吉非贝齐代谢物I对LDL氧化的抑制作用,却没有阻止atorvastatin代谢物的所述抑制作用。在atorvastatin羟基代谢物的分子结构中,羟基与该分子的羧酰胺部分相连,使这些代谢物能充当电子供体,并因此充当有效的抗氧化剂(图1)。atorvastatin邻羟基代谢物是一种比其对羟基代谢物更有效的抗氧化剂,因为氨基邻位的羟基(而不是对位的羟基)能形成一种过氧化氢自由基的相对稳定的过渡态,因此能充当有效的抗氧化剂。同样,在吉非贝齐代谢物I(而不是在吉非贝齐)中,芳香环上的羟基相当大地促成该化合物的抗氧化特性(图2)。
在氧化应激下,脂蛋白氧化涉及活性氧种类的作用。因为已知在动脉粥样硬化损伤区存在过渡金属离子,所以上述实验采用的氧化模型代表了体内的情形。
所述atorvastatin和吉非贝齐代谢物对LDL氧化的抑制作用,对VLDL和HDL也一样。在所有研究过的氧化系统中抑制模式是类似的。这些结果证明,所述代谢物通过普通机制即自由基清除和金属离子螯合来实现其对脂蛋白氧化的抑制作用。在一项研究中,对家族性高脂血症患者,吉非贝齐治疗没有显著影响LDL的可氧化性。然而,这项观察可能是因为所述药物代谢物浓度太低而不能发挥对LDL氧化的抗氧化作用,或者是因为样品收集时间不当。此外,药物代谢物能与血浆的非脂蛋白成分(如白蛋白)结合或者被隔离在细胞或间质区室内。因此,这种从所处理人或实验动物分离的脂蛋白氧化能力的离体检查,不一定反映体内脂蛋白的环境。
上面提供的数据说明,羟化胆固醇降低剂通过清除自由基和减少脂蛋白螯合金属离子而抑制脂蛋白氧化。因此,本发明提供了抑制脂蛋白氧化的方法、抑制脂蛋白螯合金属离子的方法以及清除自由基的方法。抑制脂蛋白螯合金属离子和清除自由基所需要的羟化胆固醇降低剂的剂量本文统称为“抗氧化剂量”。
所述羟化胆固醇降低剂可按抗氧化剂量即能有效引起脂蛋白氧化抑制的剂量给药。所述抗氧化有效剂量可从约1mg/kg到约100mg/kg。为了抑制脂蛋白氧化,这种剂量的活性试剂可一天给药一到四次。
所述羟化化合物可按方便的口服或胃肠外给药方式配制,并可与普通赋形剂和载体如碳酸钙、小烛树蜡、羟丙基纤维素、乳糖、硬脂酸镁、微晶纤维素、聚乙二醇、滑石和二氧化钛等结合。对口服给药,剂型可以是压成片剂或装入明胶胶囊。典型的片剂含有约10mg到约80mg活性成分。另外,所述化合物可配制成缓释给药形式,例如采用渗透泵技术以及经皮给药的皮肤贴剂。对于胃肠外给药,典型做法是,将所述化合物溶解在等渗盐溶液中用于方便的静脉内给药或注射。
Claims (7)
1.在哺乳动物中抑制脂蛋白氧化的方法,其包括施用一种抗氧化有效剂量的羟化胆固醇降低剂。
2.权利要求1的方法,其使用羟化吉非贝齐、羟化atorvastatin或羟化氟伐他汀。
3.权利要求2的方法,其使用邻位羟化或对位羟化的atorvastatin。
4.在哺乳动物中清除自由基的方法,其包括施用自由基清除剂量的羟化胆固醇降低剂。
5.权利要求4的方法,其中羟化胆固醇降低剂是邻位羟化或对位羟化的atorvastatin、羟化吉非贝齐或羟化氟伐他汀。
6.在哺乳动物中抑制脂蛋白螯合金属离子的方法,其包括施用金属离子螯合抑制剂量的羟化胆固醇降低剂。
7.权利要求6的方法,其中羟化胆固醇降低剂是邻位羟化或对位羟化的atorvastatin、羟化吉非贝齐或羟化氟伐他汀。
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Families Citing this family (39)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7250304B2 (en) * | 2000-03-31 | 2007-07-31 | The Regents Of The University Of California | Functional assay of high-density lipoprotein |
US20030040505A1 (en) * | 2000-03-31 | 2003-02-27 | The Regents Of The University Of California | Synthetic phospholipids to ameliorate atherosclerosis and other inflammatory conditions |
US20030114497A1 (en) * | 2001-07-31 | 2003-06-19 | Laman Alani | Pharmaceutical compositions of amlodipine and atorvastatin |
MX349134B (es) | 2003-01-23 | 2017-07-12 | Esperion Therapeutics Inc | Compuestos de hidroxilo y composiciones para el manejo del colesterol y usos relacionados. |
US20060194981A1 (en) * | 2005-02-26 | 2006-08-31 | Jass Paul A | Process for preparation of probucol derivatives |
WO2006104399A1 (en) * | 2005-03-26 | 2006-10-05 | Protemix Corporation Limited | Copper antagonist compositions |
EP3187182B1 (en) | 2008-09-02 | 2021-03-03 | Amarin Pharmaceuticals Ireland Limited | Pharmaceutical composition comprising eicosapentaenoic acid and nicotinic acid and methods of using same |
BRPI1011876B1 (pt) | 2009-04-29 | 2020-03-31 | Amarin Pharma, Inc. | Composições farmacêuticas estáveis compreendendo etilácido eicosapentaenoico (etil-epa) e uso das mesmas para tratar ou prevenir uma doença relacionada ao cardiovascular |
EP2424521A4 (en) | 2009-04-29 | 2015-03-04 | Amarin Pharmaceuticals Ie Ltd | PHARMACEUTICAL COMPOSITIONS COMPRISING EPA AND CARDIOVASCULAR AGENT AND METHODS OF USE |
DK2443246T3 (en) | 2009-06-15 | 2018-03-26 | Amarin Pharmaceuticals Ie Ltd | COMPOSITIONS AND METHODS FOR REDUCING TRIGLYCERIDES WITHOUT INCREASING LDL-C LEVELS IN AN INDIVIDUAL WITH CURRENT STATE THERAPY |
RU2012116079A (ru) | 2009-09-23 | 2013-10-27 | АМАРИН КОРПОРЕЙШН ПиЭлСи | Фармацевтическая композиция, включающая омега-3 жирную кислоту и гидроксипроизводное статина и способы ее применения |
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US11712429B2 (en) | 2010-11-29 | 2023-08-01 | Amarin Pharmaceuticals Ireland Limited | Low eructation composition and methods for treating and/or preventing cardiovascular disease in a subject with fish allergy/hypersensitivity |
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US11291643B2 (en) | 2011-11-07 | 2022-04-05 | Amarin Pharmaceuticals Ireland Limited | Methods of treating hypertriglyceridemia |
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WO2014074552A2 (en) | 2012-11-06 | 2014-05-15 | Amarin Pharmaceuticals Ireland Limited | Compositions and methods for lowering triglycerides without raising ldl-c levels in a subject on concomitant statin therapy |
US9814733B2 (en) | 2012-12-31 | 2017-11-14 | A,arin Pharmaceuticals Ireland Limited | Compositions comprising EPA and obeticholic acid and methods of use thereof |
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US9662307B2 (en) | 2013-02-19 | 2017-05-30 | The Regents Of The University Of Colorado | Compositions comprising eicosapentaenoic acid and a hydroxyl compound and methods of use thereof |
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US10172818B2 (en) | 2014-06-16 | 2019-01-08 | Amarin Pharmaceuticals Ireland Limited | Methods of reducing or preventing oxidation of small dense LDL or membrane polyunsaturated fatty acids |
KR101665180B1 (ko) | 2014-07-01 | 2016-10-14 | 순천향대학교 산학협력단 | 아포지단백질 a-i 돌연변이체 g59v를 포함하는 항산화성 펩타이드 및 이를 포함하는 항산화 조성물 |
EP3267989A4 (en) | 2015-03-13 | 2018-11-07 | Esperion Therapeutics, Inc. | Fixed dose combinations and formulations comprising etc1002 and ezetimibe and methods of treating or reducing the risk of cardiovascular disease |
MA41793A (fr) | 2015-03-16 | 2018-01-23 | Esperion Therapeutics Inc | Associations de doses fixes comprenant du etc1002 et une ou plusieurs statines permettant de traiter ou de réduire un risque cardiovasculaire |
US10406130B2 (en) | 2016-03-15 | 2019-09-10 | Amarin Pharmaceuticals Ireland Limited | Methods of reducing or preventing oxidation of small dense LDL or membrane polyunsaturated fatty acids |
TW201900160A (zh) | 2017-05-19 | 2019-01-01 | 愛爾蘭商艾瑪琳製藥愛爾蘭有限公司 | 用於降低腎功能下降之個體中的三酸甘油酯之組合物及方法 |
US11058661B2 (en) | 2018-03-02 | 2021-07-13 | Amarin Pharmaceuticals Ireland Limited | Compositions and methods for lowering triglycerides in a subject on concomitant statin therapy and having hsCRP levels of at least about 2 mg/L |
KR20210110890A (ko) | 2018-09-24 | 2021-09-09 | 애머린 파마슈티칼스 아일랜드 리미티드 | 대상체에서 심혈관 사건의 위험도를 감소시키는 방법 |
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US5385929A (en) * | 1994-05-04 | 1995-01-31 | Warner-Lambert Company | [(Hydroxyphenylamino) carbonyl] pyrroles |
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1998
- 1998-11-04 NZ NZ503598A patent/NZ503598A/en unknown
- 1998-11-04 US US09/485,250 patent/US6362236B1/en not_active Expired - Fee Related
- 1998-11-04 AU AU15188/99A patent/AU759376B2/en not_active Ceased
- 1998-11-04 KR KR1020057021847A patent/KR100586754B1/ko not_active IP Right Cessation
- 1998-11-04 WO PCT/US1998/023483 patent/WO1999026583A2/en not_active Application Discontinuation
- 1998-11-04 KR KR1020057014824A patent/KR20050086975A/ko not_active Application Discontinuation
- 1998-11-04 PL PL98341397A patent/PL341397A1/xx not_active Application Discontinuation
- 1998-11-04 EP EP98959378A patent/EP1047421A2/en not_active Withdrawn
- 1998-11-04 HU HU0004396A patent/HUP0004396A3/hu unknown
- 1998-11-04 TR TR2000/01522T patent/TR200001522T2/xx unknown
- 1998-11-04 CN CNB988112671A patent/CN1160068C/zh not_active Expired - Fee Related
- 1998-11-04 JP JP2000521788A patent/JP2001523694A/ja not_active Abandoned
- 1998-11-04 KR KR10-2000-7005641A patent/KR100539644B1/ko not_active IP Right Cessation
- 1998-11-04 IL IL13515498A patent/IL135154A/xx not_active IP Right Cessation
- 1998-11-04 BR BR9815039-1A patent/BR9815039A/pt not_active Application Discontinuation
- 1998-11-04 CA CA002303864A patent/CA2303864A1/en not_active Abandoned
- 1998-11-24 PE PE1998001143A patent/PE134199A1/es not_active Application Discontinuation
- 1998-11-24 CO CO98069227A patent/CO4970835A1/es unknown
- 1998-11-24 AR ARP980105959A patent/AR015487A1/es unknown
- 1998-11-24 ZA ZA9810743A patent/ZA9810743B/xx unknown
- 1998-11-24 UY UY25268A patent/UY25268A1/es not_active Application Discontinuation
- 1998-11-25 TW TW087119590A patent/TW552137B/zh not_active IP Right Cessation
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2000
- 2000-03-24 IS IS5413A patent/IS5413A/is unknown
- 2000-05-23 NO NO20002639A patent/NO20002639L/no unknown
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2001
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Also Published As
Publication number | Publication date |
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JP2001523694A (ja) | 2001-11-27 |
TW552137B (en) | 2003-09-11 |
KR100586754B1 (ko) | 2006-06-08 |
CO4970835A1 (es) | 2000-11-07 |
KR20050113288A (ko) | 2005-12-01 |
KR20050086975A (ko) | 2005-08-30 |
AR015487A1 (es) | 2001-05-02 |
KR100539644B1 (ko) | 2005-12-29 |
NZ503598A (en) | 2002-06-28 |
HK1033095A1 (en) | 2001-08-17 |
EP1047421A2 (en) | 2000-11-02 |
NO20002639D0 (no) | 2000-05-23 |
PE134199A1 (es) | 2000-01-20 |
ZA9810743B (en) | 1999-05-31 |
IS5413A (is) | 2000-03-24 |
CA2303864A1 (en) | 1999-06-03 |
IL135154A0 (en) | 2001-05-20 |
AU759376B2 (en) | 2003-04-10 |
NO20002639L (no) | 2000-05-23 |
PL341397A1 (en) | 2001-04-09 |
HUP0004396A3 (en) | 2003-04-28 |
AU1518899A (en) | 1999-06-15 |
WO1999026583A3 (en) | 1999-12-09 |
US6362236B1 (en) | 2002-03-26 |
IL135154A (en) | 2005-05-17 |
HUP0004396A2 (hu) | 2001-06-28 |
BR9815039A (pt) | 2001-03-20 |
TR200001522T2 (tr) | 2000-12-21 |
UY25268A1 (es) | 1999-07-19 |
KR20010032401A (ko) | 2001-04-16 |
CN1160068C (zh) | 2004-08-04 |
WO1999026583A2 (en) | 1999-06-03 |
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