CN1638758A - 刺激cpt-1作为一种减轻体重的方法 - Google Patents
刺激cpt-1作为一种减轻体重的方法 Download PDFInfo
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- CN1638758A CN1638758A CNA038050145A CN03805014A CN1638758A CN 1638758 A CN1638758 A CN 1638758A CN A038050145 A CNA038050145 A CN A038050145A CN 03805014 A CN03805014 A CN 03805014A CN 1638758 A CN1638758 A CN 1638758A
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Abstract
本发明提供引起体重减轻和保持最佳体重的方法和组合物,它们包括将一种刺激肉碱棕榈酰转移酶-1(CPT-1)活性的物质给予需要治疗的患者,包括人类患者。这些方法不需要抑制脂肪酸的合成。特别是,本发明提供的方法是用于开发治疗方法,此治疗方法通过药理学刺激CPT-1活性,选择性地增加脂肪酸氧化,增加能量产生和减轻肥胖,同时保持瘦肉质量。在一个优选实施方式中,以足以增加脂肪酸氧化的量给予此物质。在另一个优选实施方式中,以足以对抗丙二酸单酰辅酶A对CPT-1的抑制的量给予此物质。在再一个优选实施方式中,以足以提高丙二酸单酰辅酶A水平的量给予此物质。
Description
背景技术
本发明的领域
本发明涉及一种用于开发治疗方法的方法,所述治疗方法通过药理学刺激CPT-1活性选择性地增加脂肪酸的氧化,增加能量的产生,并减轻肥胖,同时保持瘦肉质量。
现有技术
体外MCF-7人乳腺癌细胞的浅蓝菌素治疗明显抑制脂肪酸氧化,这可能是通过丙二酸单酰辅酶A水平提高获得的(Loftus,et al.(2000)Science,288:2379-2381)。C75是α-亚甲基-γ-丁内酯家族中的一个成员,已知它们是脂肪酸合酶(FAS)的抑制剂(Kuhajda,et al.(2000)Proc.Natl.Acad Sci USA,97:3450-3454)。应用C75处理小鼠引起肝脂肪酸合成的抑制,并产生高水平的丙二酸单酰辅酶A(Loftus,et al.(2000);Pizer,et al.(2000)Cancer Res.,60:213-218)。在脑中,C75减少下丘脑神经肽-Y(NPY)的表达,导致可逆性营养不足(Loftus,et al,2000)。应用C75在体内处理ob/ob小鼠期间,尽管肝脏丙二酸单酰辅酶A水平升高,但肝脏和外周组织中脂肪出现极度的减少(Loftus,et al.,2000)。
丙二酸单酰辅酶A是一种有效的脂肪酸氧化的抑制剂,这是通过其作为肉碱-棕榈酰-转移酶-1(CPT-1)的抑制剂实现的(Witters,et al.(1992)J.Biol.Chem.,267:2864-2867)。CPT-1可使长链酰基-CoA进入脂肪酸氧化的线粒体。当用FAS抑制剂处理时,尽管通过FAS抑制引起高水平丙二酸单酰辅酶A产生,但遗传和饮食诱导的肥胖小鼠出现选择性和显著的脂肪组织丢失。因为丙二酸单酰辅酶A通过其对肉碱棕榈酰转移酶-1(CPT-1,E.C.2.3.1.21)的抑制而成为一种有效的脂肪酸氧化抑制剂,所以脂肪组织的快速和选择性减少是惊人的。期望在C75诱导营养不足过程中,系统性高水平的丙二酸单酰辅酶A可抑制脂肪酸氧化,而并不导致瘦肉质量选择性的减少。
本发明的概述
本发明的一个目的是提供引起体重减轻并保持最佳体重同时不需要抑制脂肪酸合成的方法和组合物。下面一种或多种实施方式符合这个目的和其它目的。
在一个实施方式中,本发明提供诱导体重减轻的方法,它包括将刺激肉碱棕榈酰转移酶-1(CPT-1)活性的物质(agent)给予需要治疗的患者,包括人类患者。在一个优选实施方式中,以足以提高脂肪酸氧化的量给予此物质。在另一个优选实施方式中,以足以拮抗丙二酸单酰辅酶A对CPT-1的抑制的量给予此物质。在再一个优选实施方式中,以足以提高丙二酸单酰辅酶A水平的量给予此物质。在再一个优选实施方式中,给予此物质时,丙二酸单酰辅酶A水平实质上没有升高。这里期待的丙二酸单酰辅酶A水平的实质性提高大约相当于丙二酸单酰辅酶A对CPT-1抑制作用的Ki的一半。在再一个优选实施方式中,刺激CPT-1活性的物质也抑制脂肪酸合酶(FAS)。在可选择的实施方式中,FAS没有显著抑制。这里期待的显著抑制小于15%抑制,优选小于10%抑制,更优选小于5%抑制。美国专利5,981,575中公开了测定FAS活性的方法,这里将其引用为参考文献。在上面实施方式的优选方式中,刺激CPT-1活性的物质不是具有下式的化合物:
其中R为一种取代基,它选自:
(a)含3-18个碳原子的饱和直链或支链烷基,
(b)含3-18个碳原子的不饱和直链或支链烷基,
和
其中:R1和R2相同或不同,为H、CH3、C2H5、C3H7、C4H9、CF3、OCH3、F、Cl或Br;
R3为H、CH3、C2H5、C2H5、C4H9、COOH、COOCH3、COOC2H5、COOC2H5或COOC4H9;
R4为H、CH3、C2H5、C3H7或C4H9;
X为NH、S或O;
Z为CH2、O、NH或S;
i为1至5;
j为0至10;
k为1至10;
m为1至13;和
n为1至15。
在另一个实施方式中,本发明提供一种稳定体重的方法,它包括以对FAS不显著抑制的量长期给予刺激CPT-1活性的物质。在一个优选实施方式中,以足以增加脂肪酸氧化的量给予此物质。在另一个优选实施方式中,以足以拮抗丙二酸单酰辅酶A对CPT-1的抑制的量给予此物质。在再一个优选实施方式中,以足以提高丙二酸单酰辅酶A水平的量给予此物质。在再一个优选方式中,给予此物质时,丙二酸单酰辅酶A水平实质上没有升高。这里期待的丙二酸单酰辅酶A水平的实质性提高大约相当于丙二酸单酰辅酶A对CPT-1抑制作用的Ki的一半。
在本发明的另一个实施方式中,本发明提供用于筛选引起体重减轻物质的方法,它包括确定一种候选的体重减轻物质是否刺激CPT-1活性;以及选择刺激CPT-1活性的物质。优选地,此方法还包括确定此候选的体重减轻物质是否为丙二酸单酰辅酶A对CPT-1抑制作用的拮抗剂,以及选择消除丙二酸单酰辅酶A对CPT-1抑制的候选物质。
在另一个实施方式中,本发明提供了一种治疗组合物,它包括一种刺激CPT-1活性的物质和L-肉碱。优选地,此治疗组合物包括一种丙二酸单酰辅酶A对CPT-1抑制的拮抗剂。
在另一个实施方式中,本发明还提供一个营养组合物,它包括营养上足够量的脂肪、碳水化合物和氨基酸,所述的组合物还包括一种丙二酸单酰辅酶A对CPT-1抑制的拮抗剂和L-肉碱。在一个实施方式中,此营养组合物适于胃肠外给药。
为了研究C75处理过程中肝脏丙二酸单酰辅酶A高水平情况下导致脂肪肝反常(paradoxical)减轻的作用机理,研究了C75对CPT-1活性的影响。惊奇的是,C75和相关化合物在抑制FAS的同时,伴随地刺激体外CPT-1活性和脂肪酸氧化。除了对CPT-1全面变构活化作用外,C75还消除了丙二酸单酰辅酶A对体外CPT-1活性的抑制作用。作为脂肪酸氧化增加的结果,C75提高了细胞ATP水平。
为了试验C75对体内脂肪酸氧化的作用,采用整个动物量热法测量C75处理小鼠的呼吸交换率(RER)。C75治疗后,RER在2小时内下降到0.7的范围,表明脂肪酸的氧化。这种RER的下降率类似于从用鼠粮自由喂养的动物处撤回食物。这些研究表明,尽管存在肝脏高水平的丙二酸单酰辅酶A,C75处理的动物自由地进行脂肪酸的氧化。
这些数据提示C75阻断了丙二酸单酰辅酶A对体内CPT-1活性的抑制作用,导致FAS抑制期间脂肪肝和脂肪质量的减少。本发明描述了一种开发治疗方法的方法,所述治疗方法通过CPT-1活性的药理学刺激,选择性地减轻肥胖,同时保持瘦肉质量。
附图的简要说明
附图1显示在MCF-7细胞中C75对脂肪酸氧化的作用,与乙莫克舍的作用相比较。
附图2显示C75对CPT-1活性的浓度依赖性刺激作用和丙二酸单酰辅酶A引起的抑制作用。
附图3显示C75对CPT-1的可逆性刺激作用。
附图4显示不同C75类似物对CPT-1的刺激作用。
附图5显示在MCF-7细胞中C75引起的细胞ATP水平的浓度依赖性提高。
附图6显示在小鼠脂肪细胞中C75对脂肪酸氧化的浓度依赖性刺激作用。
附图7显示在小鼠脂肪细胞中C75引起的细胞ATP水平的浓度依赖性提高。
附图8显示间接量热法测定的小鼠呼吸交换率(RER),(A)为没有C75的结果,(B、C)为C75存在下的结果。
优选实施方式的详细描述
已经表明体内脂肪酸合酶的抑制可引起快速而显著的体重减轻。浅蓝菌素为一种天然产物,C-75为一种合成小分子,当将它们经脑室内(i.c.v.)应用于大鼠时,两者均可引起类似的体重减轻。当进行全身治疗(例如腹膜内给药)时,C-75引起更显著的体重减轻,甚至体重减轻大于饥饿的动物。这些数据表明C-75对体重减轻的外周(非CNS)作用大于浅蓝菌素。
在研究C-75这种显著外周作用的机理的同时,本发明人最近发现,除了FAS的抑制外,C-75及其α-亚甲基-γ-丁内酯家族直接刺激肉碱棕榈酰转移酶-1(CPT-1),导致线粒体脂肪酸氧化的增加。相反,通过FAS抑制引起高水平丙二酸单酰辅酶A的产生,浅蓝菌素导致CPT-1活性的降低和脂肪酸氧化的减少。
体外MCF-7细胞的C75处理刺激CPT-1活性从150至160%。同时也伴随脂肪酸氧化的增加。在C75类似物中,对CPT-1的刺激活性来说,C6-C16的碳链长度最佳。在C75存在的情况下,丙二酸单酰辅酶A不再能抑制CPT-1的活性,这提示除了其刺激作用外,C75还阻止了丙二酸单酰辅酶A对CPT-1的抑制作用。在CPT-1和C75之间没有发现可检测到的共价相互作用。
因此,C75的外周性(非CNS)体重减轻作用至少部分是由于CPT-1的刺激作用和脂肪酸氧化的增加,同时伴随脂肪酸合成的抑制。这些数据将α-亚甲基-γ-丁内酯家族等同于丙二酸盐或丙二酸单酰辅酶A模拟物,并将CPT-1确定为体重减轻疗法中的靶位。更广泛的是,我们的数据提示,可设计并合成其它的丙二酸盐或丙二酸单酰辅酶A模拟物,使其成为有效的体重减轻物质。
数据表明C-75及其α-亚甲基-γ-丁内酯家族直接与CPT-1相互作用,导致CPT-1酶活性和脂肪酸氧化的增加。美国专利5,981,575公开了C75的化学结构和众多的类似物,以及合成这些类似物的方法,此专利在此引用为参考文献。C75的刺激作用与饱和碳侧链的长度有关,其最佳长度为6至18个碳原子。关于本发明的讨论,C75是刺激CPT-1的原型物质;下文中提及的C75包括其它刺激CPT-1活性的适宜物质,除了上下文另作说明。
除了直接作用于CPT-1外,C-75还消除了丙二酸单酰辅酶A对CPT-1活性的抑制作用。尽管C75显示出与纯化FAS(1)的慢结合抑制剂的动力学特征,但它与CPT-1的相互作用却是快速而有竞争性的。因此,C75对脂肪酸氧化的刺激作用或是由于其对CPT-1活性的直接刺激作用,其对丙二酸单酰辅酶A抑制CPT-1的干扰,或是由于两者。令人感兴趣的是,C75的作用并不限于鼠CPT-1,它对人CPT-1也有类似的作用。作为脂肪酸氧化增加的结果,C75也提高了人和鼠细胞中ATP的水平。
C75对体内脂肪酸代谢的作用反映为细胞水平上可见的变化。尽管C75引起体内高水平丙二酸单酰辅酶A的产生,但C7 对瘦鼠的处理导致脂肪酸氧化极度而快速的增加。因此,C75及其α-亚甲基-γ-丁内酯家族似乎充当了CPT-1的竞争性激动剂。C75的这种激动剂活性似乎克服了丙二酸单酰辅酶A对同一种酶的抑制作用。C75所致脂肪酸氧化的增加是C75治疗小鼠过程中肥胖显著减轻的一个重要机理。
总之,本发明描述了一种开发治疗方法的方法,此治疗方法通过药理学刺激CPT-1活性,选择性地增加脂肪酸的氧化,增加能量的产生,并减轻肥胖,同时保持瘦肉质量。
含有C75和/或刺激CPT-1的其它物质的治疗组合物的制剂,以及应用这些物质的方法是在本领域技术范围内,特别考虑到美国专利5,981,575的说明书,此专利文本在此引用为参考文献。
通过在给予脂肪酸或含有脂肪酸残基的化合物的同时给予CPT-1刺激剂,而使用CPT-1刺激剂增加能量的产生也在本领域的技术范围内,特别考虑到美国专利4,434,160中公开的营养组合物,此专利文本在此引用为参考文献。
实施例
为了使本发明得到更充分的理解,下面提供了一些实施例。但是,并不是将本发明的范围限制在这些实施例中的特殊实施方式中,它们仅是用来进行举例说明。
实施例1脂肪酸合酶抑制剂的反常作用
浅蓝菌素是一种FAS抑制剂,它增加MCF-7细胞中丙二酸单酰辅酶A的量(3)。丙二酸单酰辅酶A的大量增加,结果通过丙二酸单酰辅酶A对CPT-1的抑制作用,使浅蓝菌素引起脂肪酸氧化的抑制(Thupari,et al.(2001)Biochem.Biophys.Res.Comm.,285:217-223)。在前面已经表明,C75处理MCF-7细胞,通过C75对脂肪酸合酶(FAS)的抑制,导致丙二酸单酰辅酶A5倍以上的增加(3)。C75对脂肪酸氧化的作用试验如下。
由American Type Culture Collection获得人乳腺癌细胞系MCF-7。将1×106MCF-7细胞铺在T-25瓶中,一式三份,并在37℃孵育过夜。然后按照所示加入物质,从5mg/ml含原料的DMSO稀释。2小时后,除去含药物的培养基,用1.5ml下面的缓冲液将细胞预孵育30分钟:114mM NaCl、4.7mM KCl、1.2mM KH2PO4、1.2mM MgSO4、葡萄糖11mM。预孵育后,加入200μl分析缓冲液,它含有114mM NaCl、4.7mM KCl、1.2mM KH2PO4、1.2mM MgSO4、葡萄糖11mM、2.5mM与白蛋白结合的棕榈酸盐(含有10μCi[1-14C]棕榈酸盐)、0.8mM L-肉碱,并在37℃将细胞孵育2小时。孵育后,将400μl苄索氯铵加入中心孔中,以收集释放的14CO2。即刻,将500μl 7%高氯酸加入细胞中,使反应停止。然后在37℃下将此带孔的烧瓶孵育2小时,随后除去苄索氯铵,并计算14C。空白的制备是在用分析缓冲液孵育2小时前,将500μl 7%高氯酸加入这些细胞中而获得。
用C75处理细胞2小时,然后测定脂肪酸氧化,与对照组相比,C75处理导致脂肪酸氧化156%的增加(见附图1;p=0.0012,双尾(two-tailed)t检验,Prism 3.0)。乙莫克舍是一种已知的脂肪酸氧化抑制剂和CPT-1非竞争性抑制剂,相反,它将脂肪酸氧化降低至对照组的32%(p=0.0006,双尾t检验,Prism 3.0)。在剂量为5-20μg/ml情况下,C75对MCF-7细胞的处理重复引起脂肪酸氧化的增加。
反常的是,尽管丙二酸单酰辅酶A的增加与浅蓝菌素引起的增加相似,但C75却增加了MCF-7细胞中脂肪酸的氧化,而不是降低了脂肪酸的氧化。这种情况提示C75对肉碱棕榈酰转移酶-1(CPT-1)具有直接作用。
实施例2 C75刺激人CPT-1的活性
通过下面步骤测定MCF-7细胞中CPT-1活性:在24孔平板上,将106MCF-7细胞平铺在含10%胎牛血清的DMEM中,一式三份。在37℃孵育过夜后,除去此培养基,并用700μl分析培养基代替,分析培养基由下列组成:50mM咪唑、70mM KCl、80mM蔗糖、1mM EGTA、2mM MgCl2、1mM DTT、1mM KCN、1mM ATP、0.1%脂肪酸游离牛血清白蛋白、70μM棕榈酰-CoA、0.25μCi[甲基-14C]L-肉碱、40μg洋地黄皂甙,含或不含20μM丙二酸单酰辅酶A。在37℃孵育6分钟后,通过加入500μl冰冻4M高氯酸而停止反应。然后收获细胞,以13,000×g离心5分钟。用500μl冰冻2mM高氯酸冲洗粒状沉淀,然后再次离心。将所得粒状沉淀再悬浮在800μl dH2O中,并用150μl丁醇提取。通过液体闪烁计数法对此丁醇相计数,此丁醇相代表酰基肉碱衍生物。
用10或20μg/mL的C75处理MCF-7细胞1小时,然后测定CPT-1活性。用C75和标示的丙二酸单酰辅酶A浓度(“M”表示20uM的丙二酸单酰辅酶A)进行此分析。单独的丙二酸单酰辅酶A处理导致CPT-1活性46%的下降,这与前面实验相似(见附图2;p=0.02,双尾t检验,Prism 3.0)。丙二酸单酰辅酶A对CPT-1活性的抑制水平与MCF-7细胞中CPT-1的肝脏同工型的活性一致。对于CPT-1的肝脏同工型,丙二酸单酰辅酶A的Ki约为7μM,而对于CPT-1的肌肉同工型,其Ki为0.07μM。因此,MCF-7细胞主要表达CPT-1的肝脏同工型(这与免疫印染法一致)。
C75或C75和丙二酸单酰辅酶A处理细胞之间CPT-1活性的差异没有统计学意义(附图2)。因此,在C75存在情况下,丙二酸单酰辅酶A丧失了其对CPT-1的抑制作用;相反,不管丙二酸单酰辅酶A存在与否,C75都产生对CPT-1的刺激作用。因此,在C75存在情况下,丙二酸单酰辅酶A正常的抑制活性丧失了。丙二酸单酰辅酶A对CPT-1活性的抑制表明,C75及相关的化合物是激活了CPT-1活性,而不是CPT-2活性,丙二酸单酰辅酶A不能抑制CPT-2活性。
在随后的实验中(数据见附图3),在测定CPT-1活性前,不处理MCF-7细胞(左侧柱图)或者用20μg/mL C75处理MCF-7细胞1小时(中间和右侧柱图)。在测定CPT-1的6分钟过程中,将C75从分析缓冲液中除去,并用缓冲液代替(中间柱图),或者加入20μM丙二酸单酰辅酶A(左侧和右侧柱图)。在分析过程中,单独丙二酸单酰辅酶A处理导致CPT-1活性大约70%抑制(左侧柱图)(p=0.0045,双尾t检验,Prism 3.0)。前面分析缓冲液中不含C75的C75处理导致CPT-1活性为对照组的158%(p=0.028,双尾t检验,Prism 3.0),这与C75在分析缓冲液中时的结果相似(见上面实验)。但是,当从反应缓冲液中除去C75,并用丙二酸单酰辅酶A代替时,C75的刺激活性就丧失了(右侧柱图)。因此,C75没有与CPT-1存在可检测的共价结合,而且C75可能是丙二酸单酰辅酶A的竞争性拮抗剂。这些数据也提示,C75在丙二酸单酰辅酶A的结合位点上与CPT-1相互作用。
实施例3有效的CPT-1刺激物的结构
按照美国专利5,981,575所述的方法制备仅在饱和碳“尾部”长度上不同的α-亚甲基-γ-丁内酯类似物,此专利在此引用为参考文献。C75具有一个8碳尾部,C12和C16的尾部分别具有12个和16个碳。在测定CPT-1活性前,用20μg/ml的C75和C75类似物处理细胞1小时。因为整个细胞对丙二酸单酰辅酶A是不渗透的,所以丙二酸单酰辅酶A仅被加入反应缓冲液中。C75在20μg/ml下将CPT-1活性刺激到对照组的166%(见附图4;p=0.0092,双尾t检验,Prism 3.0)。C12类似物将CPT-1活性刺激到对照组的186%(p=0.0099,双尾t检验,Prism 3.0),而C16类似物将CPT-1活性刺激到对照组的138%(p=0.055,双尾t检验,Prism 3.0)。丙二酸单酰辅酶A是CPT-1的细胞内竞争性抑制剂,它在20μM下将CPT-1活性降低到对照组的64%(p=0.023,双尾t检验,Prism 3.0)。对于CPT-1活化,最佳的碳链长度在6和16碳之间。
实施例4 CPT-1刺激引起的脂肪酸氧化的增加产生ATP
C75处理后,脂肪酸氧化的增加导致MCF-7细胞中ATP升高。将1×105MCF-7细胞平铺在96孔平板上。用C75或赋形剂处理细胞。2小时后,采用ATP Bioluminescence Kit CLS II(Roche),按照厂商的方案应用虫荧光素酶测定法测定ATP。通过Perkin Elmer Wallac Victor21420发光计读这些平板。10μg/ml和20μg/ml的C75处理均导致细胞ATP总量的明显升高(见附图5;p=0.0001;与对照组相比p<0.0001,双尾t检验,Prism 3.0)。用C75孵育1小时后,也获得相似的结果。因此,C75使脂肪酸氧化增加导致细胞产生能量的增加。
实施例5 C75刺激肌肉型CPT-1的活性
为了将C75对脂肪酸代谢作用的研究超出癌细胞系,并扩展到正常脂肪细胞上,在应用MCF-7细胞类似的分析中使用分化的(非转化的)鼠NIH 3T3-L1脂肪细胞。从American Type Culture Collection处获得3T3-L1细胞,并在含高葡萄糖(4.5g/l)(Gibco 12100-046)的DMEM中,用10%胎牛血清和0.008g/L Biotin(Sigma B-4639)培养这些细胞。这些细胞融合后3天引发分化,同时用分化培养基取代标准培养基。分化培养基包括标准培养基,为获得最终的浓度,其中加入了下列物质:甲基异丁基黄嘌呤(MIX)0.5mM、地塞米松(DEX)1μM和胰岛素10μg/ml。48小时后,用后分化培养基取代分化培养基,后分化培养基包含上述浓度的胰岛素,不含MIX和DEX。在7至10天中这些分化细胞可以用于实验。
C75增强了分化为脂肪细胞的NIH 3T3-L1细胞中CPT-1活性和脂肪酸代谢。后分化1周,或用对照赋形剂,或用C75处理细胞2小时,所用剂量如下。按照实施例2、1和4中所述的方法测定CPT-1活性、脂肪酸氧化和细胞ATP总量。用C75处理3T3-L1脂肪细胞导致CPT-1活性的增加为对照组的377%(p<0.0001,双尾t检验,Prism 3.0)。剂量为20μg/ml或以上的C75引起CPT-1活性的增加,结果导致脂肪酸氧化的显著增加(见附图6;20μg/ml,p=0.007;<20μg/ml,p<0.0001;双尾t检验,Prism 3.0)。而且,在C75的剂量为20μg/ml或以上时,脂肪酸氧化的增加导致ATP水平的显著增加(见附图7;20μg/ml,p=0.05;30μg/ml,p<0.01;40μg/ml,p<0.0001;双尾t检验,Prism3.0)。C75导致的脂肪酸氧化的增加可能引起C75给药时发现的体内脂肪组织质量明显减少。
实施例6 CPT-1的体内刺激将代谢转变为脂肪酸氧化
与C75对人和鼠CPT-1和脂肪酸代谢的作用相一致,C75显著而快速的刺激体内脂肪酸氧化。将小鼠保持在Oxymax量热器中(OxymaxEqual Flow System,Columbus Instrument s,Columbus,OH)。使用间接量热器同时测定至多4个小鼠氧的消耗和CO2的产生。在整个实验过程中,每15分钟记录一次测定值。通过版本5.9的Oxymax软件计算呼吸交换率(RER)。RER的定义是任意给定时间的CO2与O2的比率,不管平衡是否达到。保持小鼠可以自由获得水和鼠粮。在对照小鼠中(附图8A),观察RER昼间变化以显示这些动物的进食和休息周期。RER为1,与碳水化合物的氧化一致,而0.7显示脂肪酸的氧化。用C75处理小鼠,并将它们保持在Oxymax量热器中,这样显示呼吸交换率(RER)快速下降到大约0.7(附图8B)。30mg/kg的C75处理破坏了对照小鼠昼间方式,表现为在约2小时内RER的快速下降以完成脂肪酸氧化。类似的,以20mg/kg的C75处理,RER显示出相似的下降速率,但没有延长的作用(附图8C)。重要的是,RER的下降速率与禁食动物中观察到的(数据未显示)相似。
尽管C75引起体内丙二酸单酰辅酶A水平提高,但如RER的测定所示,C75处理导致脂肪酸氧化快速而显著的增加。因此,尽管在体内FAS抑制过程中产生高水平的丙二酸单酰辅酶A,通过使脂肪酸氧化发生,C75处理的动物可明显减少脂肪质量并可逆转脂肪肝。
尽管为了清晰理解本发明,前面通过例证和实施例对本发明某些细节进行了描述,但显而易见,在附加的权利要求范围内可对本发明进行某些改变和修改。对于医疗、免疫学、杂交瘤技术、药理学和/或相关领域的技术人员,为实现本发明而对上述方式进行的修改应当在下面权利要求的范围内。
本发明书中提及的所有出版物和专利申请是用来表示本发明所属领域中技术人员的技术水平。所有这些出版物和专利申请在此均同等地收录为参考文献,好象每个单个出版物或专利申请均特意和单独地指明被引用为参考文献。
Claims (5)
1、引起体重减轻的方法,其包括给予一种刺激肉碱棕榈酰转移酶-1(CPT-1)活性的物质。
2、稳定体重的方法,其包括以不明显抑制FAS的量长期给予一种刺激CPT-1活性的物质。
3、筛选引起体重减轻的物质的方法,其包括确定候选的体重减轻物质是否刺激CPT-1活性,以及筛选刺激CPT-1活性的物质。
4、治疗组合物,其包括一种刺激CPT-1活性的物质和L-肉碱。
5、营养组合物,其包括营养上足够量的脂肪、碳水化合物和氨基酸,所述组合物还包括L-肉碱和一种丙二酸单酰辅酶A抑制CPT-1的拮抗剂。
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2006
- 2006-10-02 US US11/537,968 patent/US7459481B2/en not_active Expired - Fee Related
-
2008
- 2008-11-06 US US12/266,425 patent/US20090124684A1/en not_active Abandoned
- 2008-11-20 AU AU2008249144A patent/AU2008249144A1/en not_active Abandoned
-
2009
- 2009-10-20 JP JP2009241309A patent/JP2010047594A/ja active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1924011B (zh) * | 2005-08-29 | 2011-05-11 | 弗·哈夫曼-拉罗切有限公司 | Cpt-2晶体 |
Also Published As
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US20070087037A1 (en) | 2007-04-19 |
EP1471906A4 (en) | 2006-02-01 |
WO2003066043A1 (en) | 2003-08-14 |
EP1471906A1 (en) | 2004-11-03 |
KR20040082417A (ko) | 2004-09-24 |
AU2003215111A1 (en) | 2003-09-02 |
EA200401052A1 (ru) | 2005-02-24 |
US20050143467A1 (en) | 2005-06-30 |
CA2474884A1 (en) | 2003-08-14 |
MXPA04007556A (es) | 2005-12-05 |
US20050106217A1 (en) | 2005-05-19 |
CN1313089C (zh) | 2007-05-02 |
AU2008249144A1 (en) | 2008-12-11 |
US7459481B2 (en) | 2008-12-02 |
JP2010047594A (ja) | 2010-03-04 |
EA007029B1 (ru) | 2006-06-30 |
US20090124684A1 (en) | 2009-05-14 |
BR0307421A (pt) | 2004-12-28 |
JP2005523270A (ja) | 2005-08-04 |
IL163312A (en) | 2011-12-29 |
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