CN110461864B - 具有抵抗环境污染物质的细胞保护效果的肽及其用途 - Google Patents
具有抵抗环境污染物质的细胞保护效果的肽及其用途 Download PDFInfo
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Abstract
本发明涉及具有抵抗环境污染物质的细胞保护功效的肽及其用途,确认由SEQ ID NO:1、2或3的氨基酸序列组成的肽与被知为二噁英系列中毒性最强的2,3,7,8‑四氯二苯并二噁英(以下称为TCDD)直接结合(binding)而防止皮肤内渗透,发挥防止由TCDD及微细尘内包含的多环芳族烃导致的AhR活性机理的作用,这样的直接的抵抗环境污染物质的细胞保护效果与间接阻碍对于物质的接触机会或通过肠壁强化而使毒性减小的以往的方法具有差别性。
Description
【技术领域】
本发明涉及具有抵抗环境污染物质的细胞保护功效的肽及其用途。
【背景技术】
环境污染物质被分类为所谓内分泌系障碍物质,其种类包含二噁英的有机氯系物质,农药,邻苯二甲酸盐,苯并芘,五-壬基苯酚,双苯酚A,重金属,苯乙烯二聚体及三聚体等。这些物质化学上很稳定而在环境中残留的时间长,活体蓄积度高,通过食物链可达人体。
其中,二噁英是氯化烃化合物,常规是指具有相互近似的化学结构的二噁英类(多氯化二苯并-p-二噁英(polychlorinated dibenzo-p-dioxin);PCDD)和呋喃类(polychlorinated dibenzofuran;PCDF),根据被取代的氯的数而存在75种的PCDD和135种的PCDF异构体。
主要通过垃圾焚烧,纸或纸浆的漂白过程,塑料制造,有机氯系农药的制造等排出或生成。在常温下是无色的结晶性固体,由于热化学稳定,易溶于脂肪而具有进入生物体时蓄积在脂肪组织的性质。对人体的影响具有生殖功能障碍,乳酸,胚胎的非正常发达,激素调节功能的变化,糖尿病发病,免疫体系的异常,癌诱发可能性等。
微细尘由亚硫酸气体,氮氧化物,铅,臭氧,一氧化碳等组成,将颗粒的尺寸在直径10μm以下的微细的尘称作PM10,在颗粒的直径在2.5μm以下时称作超微细尘PM2.5。微细尘的自然发生原因有土尘,在海水形成的盐,植物的花粉等,人为发生原因有燃烧化石燃料等时生成的煤烟,汽车排出气体,建筑现场的扬尘,工厂内粉末形态的原材料,副材料,而且,焚烧场烟等。
这样的微细尘的颗粒的直径是人头发粗度的1/5~1/7左右,很小,不被人体的鼻腔,口腔,器官过滤而深度渗透。以与所述的有害物质吸附的形态进入身体内而诱发气管,肺,心血管,脑等各器官的炎症反应而诱发呼吸器官疾病,心血管系疾病。另外,2013年10月被世界卫生组织下的国际癌症研究所分类为1组致癌物质。
已知二噁英和微细尘内包含的多环芳族烃(polycyclic aromatic hydrocarbon,PAH)渗透到细胞内而共同使芳基烃受体(Aryl hydrocarbon receptor;AhR)活化。通过AhR活化机理而细胞内活性氧(Reactive oxygen species;ROS)增加,AhR、ARNT 2种二聚体复合体(heterodimer complex)与核内异生素反应要素(xenobiotic response element;XRE)结合而增加多样的炎症介导因子,使作为黑色素合成关联转录因子的小眼球症关联转录因子(Microphthalmia-associated transcription factor;Mitf),作为皱纹生成关联酶的基质金属蛋白酶(Matrix metalloproteases;MMPs)表达而在皮肤引发皮肤炎,色素过形成(色斑,雀斑等),皱纹等。
以往可用的抗污染产品利用电荷防止微细尘的皮肤吸附或使用使皮肤肠壁强化的原料。这呈现不防止二噁英或微细尘这样的环境污染物质的直接的作用机理的间接的保护效果。在环境污染物质直接接触皮肤而影响皮肤细胞时,但尚未开发阻碍其的原料或产品。
【发明详述】
【技术课题】
对此,本发明人确认由SEQ ID NO:1、2或3的氨基酸序列组成的肽与已知在二噁英系列中毒性最强的2,3,7,8-四氯二苯并二噁英(以下称为TCDD)直接结合(binding)而防止皮肤内渗透,发挥防止由TCDD及微细尘内包含的多环芳族烃导致的AhR活性机理的作用。
对此,本发明的目的是提供由SEQ ID NO:1、2或3的氨基酸序列组成的肽。
本发明的别的目的是提供由二噁英类似物质诱发的疾病的预防或治疗用药物组合物,其包含选自由SEQ ID NO:1、2或3的氨基酸序列组成的肽的1种以上。
本发明的再别的目的是提供由二噁英类似物质诱发的疾病的缓和和/或改善用食品组合物,其包含选自由SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽的1种以上的肽作为有效成分。
本发明的别的目的是提供皮肤状态改善用化妆品组合物,其包含选自由SEQ IDNO:1、2或3的氨基酸序列组成的肽的1种以上。
本发明的再别的目的是提供由二噁英类似物质诱发的疾病的预防或治疗方法。
本发明的再别的目的涉及由SEQ ID NO:1、2或3的氨基酸序列组成的肽用于由二噁英类似物质诱发的疾病的预防或治疗用途。
【课题解决手段】
本发明涉及具有抵抗环境污染物质的细胞保护功效的肽及其用途,确认由SEQ IDNO:1、2或3的氨基酸序列组成的肽与被知为二噁英系列中毒性最强的2,3,7,8-四氯二苯并二噁英(以下称为TCDD)直接结合(binding)而防止皮肤内渗透,发挥防止由TCDD及微细尘内包含的多环芳族烃导致的AhR活性机理的作用,这样的直接的抵抗环境污染物质的细胞保护效果与间接阻碍与物质的接触机会或通过肠壁强化而使毒性减小的以往的方法具有差别性。
接下来更详细地说明本发明。
本发明的一实施方式涉及由SEQ ID NO:1、2或3的氨基酸序列组成的肽。
为了选定氨基酸序列的一部分部位并使其活性增加,所述肽可为诱导了N-末端和/或C-末端变形的。通过这样的N-末端和/或C-末端变形可显著地提高本发明的肽的稳定性,例如,在肽的活体内施用时可使半衰期增加。
所述N-末端变形可为在肽的N-末端结合选自乙酰基(acetyl group),芴基甲氧基羰基(fluoreonylmethoxycarbonyl group),甲酰基(formyl group),棕榈酰基(palmitoylgroup),肉豆蔻基(myristyl group),硬脂基(stearyl group)及聚乙二醇(polyethyleneglycol;PEG)的保护基。所述保护基发挥保护本发明的肽免于活体内的蛋白切断酶的攻击的作用。
所述C-末端变形可为在肽的C-末端结合了羟基(hydroxyl group,-OH),氨基(amino group,-NH2),叠氮基(azide,-NHNH2)等的,但不限于此。
根据本发明的一实施方式,由SEQ ID NO:1、2或3的氨基酸序列组成的肽与被知为二噁英系列中毒性最强的2,3,7,8-四氯二苯并二噁英(以下称为TCDD)直接结合(binding)而防止皮肤内渗透,呈现防止由TCDD及微细尘内包含的多环芳族烃导致AhR活性机理的效果。
流入细胞内的多环芳族烃与AhR结合而向核内移动,与AhR核转位蛋白(AHRnucleartranslocator,ARNT)形成复合体,其与二噁英反应元件(dioxin-responsiveelement,DRE)结合以促进下方基因的表达。下方基因有CYP1A1及诸炎症关联因子。CYP1A1与苯并芘这样的诸致癌前体因子的代谢关联,已知这样生成的代谢物与DNA反应而引发突变。另外,还增加COX2,TNF-a及IL-1b这样的炎症关联因子的表达而诱发接触部位及全身的炎症反应。
这样的结果表明本发明的肽对于由二噁英类似物质诱发的疾病的预防和/或治疗具有很优秀的功效。
本发明的别的一实施方式涉及由二噁英类似物质诱发的疾病预防或治疗用药物组合物,其包含选自由SEQ ID NO:1、2或3的氨基酸序列组成的肽的1种以上的肽。
所述由二噁英类似物质诱发的疾病非常复杂,可包括氯性痤疮这样的皮肤疾病,***数减小,睾丸癌,***癌,子宫内膜增殖症,乳腺癌,肝毒性,免疫功能低下,高脂血症,尿道下裂,隐睾症,畸形儿出生,血管损伤,肝细胞性癌,肝的肥大,腺纤维症,体重损失,毛发损失,口腔浮肿,眼睑浮肿或胃粘膜性溃疡等,但不限于此(Carter et al.,Science188:738(1975);Fourth Annual Report on Carcinogens,NTP 85-002:170,185(1985))。
本说明书的术语“二噁英”是一般在大众媒体使用的术语,作为2,3,7,8-四氯二苯并二噁英(以下称为TCDD)的缩略语被使用。但是,TCDD是多氯化二苯并-p-二噁英(以下称为PCDD)家族的1种,PCDD根据氯原子的位置及数而具有75种的同族体(congener)。在生物学上,TCDD被知为具有最强力的毒性的PCDD。
另一方面,已公知与TCDD具有共同的生物学特性的别的芳族烃,例如,有多氯化二苯并呋喃(PCDFs)家族及多氯化联苯(PCBs)家族。
对此,作为在本说明书中使用的术语的“二噁英”是指PCDD内包含的全部化合物,“二噁英类似物质”包含所述的PCDD,PCDFs及PCBs,是指呈现与PCDD相同的细胞效果的物质。
二噁英作为致癌性物质而被报告诱发生殖***及发育过程扰乱,免疫体系的损伤及激素调节作用的干涉等的严重的疾病(Yang,J.H.et al.,Carcinogenesis.20:13-18(1999),Lee,Y.W.et al.,Toxicol.Lett.,102-103:29-83(1998))。
根据美国环境署(EPA)的报告,作为对于在二噁英及二噁英类似物质漏出时候发生的人体内的不良影响的统计,报告了与50年前比较而男性的***数减小至50%水平,睾丸癌发生频度增加3倍左右,***癌增加2倍以上。另外确认,由二噁英等导致与过去不同地子宫内膜增殖症和女性的乳腺癌发生频度的增加等。
所述药物组合物可包含药剂学有效量的选自由SEQ ID NO:1、2或3的氨基酸序列组成的肽的1种以上的肽。
另外,所述药物组合物可还包含药剂学容许的载体。
所述药剂学容许的载体是在制剂时常规利用的,包含乳糖,右旋糖,蔗糖,山梨糖醇,甘露糖醇,淀粉,金合欢橡胶,磷酸钙,藻酸盐,明胶,硅酸钙,微晶性纤维素,聚乙烯吡咯烷酮,纤维素,水,糖浆,甲基纤维素,甲基羟基苯甲酸酯,丙基羟基苯甲酸酯,滑石粉,硬脂酸镁及矿物油等,但不限于此。适合的药剂学容许的载体及制剂详细记载在Remington'sPharmaceutical Sciences(19th ed.,1995)。
本发明的药物组合物除了上述成分之外,可还包含润滑剂,湿润剂,甜味剂,香味剂,乳化剂,悬浮剂,保存剂等,但不限于此。
所述药物组合物可经口或非经口,优选非经口施用,在非经口施用时,可通过肌肉注入,静脉内注入,皮下注入,腹腔注入,局部施用,经皮施用等施用,但不限于此。
所述药物组合物的施用量可为每人0.0001~1000μg(微克),0.001~1000μg,0.01~1000μg,0.1~1000μg,或者1.0~1000μg,但不限于此,可根据制剂化方法,施用方式,患者的年龄,体重,性别,病理状态,饮食,施用时间,施用途径,***速度及反应感应性这样的因素多样地处置。
所述药物组合物可根据本发明所属领域的普通技术人员可容易实施的方法,利用药剂学容许的载体和/或赋形剂而制剂化而制造为单位容量形态或放入多容量容器内而制造。
所述剂型可为油或水性介质中的溶液,悬浮液或乳液形态,还可为提取剂,粉末剂,颗粒剂,片剂或胶囊剂形态,可还包含分散剂和/或稳定化剂。
本发明的再别的一实施方式涉及对二噁英类似物质具有拮抗性的食品组合物,其包含选自由SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽的1种以上的肽作为有效成分。
本发明的再别的一实施方式涉及由二噁英类似物质诱发的疾病的缓和和/或改善用食品组合物,其包含选自由SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽的1种以上的肽作为有效成分。
所述由二噁英类似物质诱发的疾病可包括:***数减小,睾丸癌,***癌,子宫内膜增殖症,乳腺癌,肝毒性,免疫功能低下,高脂血症,尿道下裂,隐睾症,畸形儿出生,血管损伤,肝细胞性癌,肝的肥大,腺纤维症,体重损失,毛发损失,口腔浮肿,眼睑浮肿或胃粘膜性溃疡等,但不限于此。
所述食品可为各种食品,饮料,食品添加剂等。
作为所述食品组合物中所含有的有效成分的肽的含量可根据食品的形态,期望的用途等而适宜,无特别的限制,例如,可以全体食品重量的0.01~15重量%添加,健康饮料组合物可以100ml为基准,以0.02~10g,优选以0.3~1g的比率添加。
在所述食品是饮料时,除了作为必需成分而以指示的比率含有所述肽之外,液体成分中无特别的限制点,如常规的饮料一样可作为追加成分含有多种香味剂或天然碳水化合物等。
所述天然碳水化合物是单糖,例如,葡萄糖,果糖等的二糖,例如,麦芽糖,蔗糖等,及多糖,例如,糊精,环糊精等这样的常规的糖及木糖醇,山梨糖醇,赤藓糖醇等的糖醇。
作为上述之外的香味剂,可有利地使用天然香味剂(祝马丁),甜叶菊提取物(例如,甜叶菊苷A,甘草酸等)及合成香味剂(糖精,阿司帕坦等)。所述天然碳水化合物的比率是每100ml本发明的组合物一般约1~20g,优选约5~12g。
除了上述之外,本发明的食品组合物可含有多种营养剂,维生素,矿物(电解质),合成香料及天然香料等的香料,着色剂及增量剂(干酪,巧克力等),果胶酸及其盐,藻酸及其盐,有机酸,保护性胶体增粘剂,pH调节剂,稳定化剂,防腐剂,甘油,醇,碳酸饮料中使用的碳酸化剂等。
此外,本发明的食品组合物可含有用于制造天然水果汁及水果汁饮料及野菜饮料的果肉。这样的成分可独立地或组合使用。这样的添加剂的比率虽不那么重要,但一般从每100重量份本发明的组合物0~约20重量份的范围选择。
本发明的再别的实施方式涉及皮肤状态改善用化妆品组合物,其包含选自由SEQID NO:1、2或3的氨基酸序列组成的肽的1种以上。
所述化妆品组合物可包含:(a)化妆品学有效量(cosmetically effectiveamount)的上述本发明的肽;和/或(b)化妆品学容许的载体,但不限于此。
在本说明书中,术语“化妆品学有效量”是指对于达成上述本发明的组合物的皮肤状态改善功效而言充分的量。
所述化妆品组合物也可制造为在本领域常规制造的任何剂型,可剂型化为例如,溶液,悬浮液,乳液,膏剂,凝胶,霜,洗液,粉末,皂,表面活性剂-含有清洁,油,粉底,乳液底料,蜡底料及喷雾剂等,但不限于此。更详细地,可制造为软化化妆水,营养化妆水,营养霜,按摩霜,精华液,眼霜,清洁霜,清洁泡沫,清洁水,包装,喷雾剂或粉末的剂型。
在本发明的化妆品的剂型是膏剂,霜或凝胶时,作为载体成分,可利用动物性油,植物性油,蜡,石蜡,淀粉,黄蓍胶,纤维素衍生物,聚乙烯二醇,硅,膨润土,氧化硅,滑石或氧化锌等。
在本发明的化妆品的剂型是粉末或喷雾剂时,作为载体成分,可利用乳糖,滑石,氧化硅,氢氧化铝,硅酸钙或聚酰胺粉末,尤其是在喷雾剂时,可还包含氯氟烃,丙烷/丁烷或二甲基醚这样的推进剂。
在本发明的化妆品的剂型是溶液或乳液时,作为载体成分,可利用溶剂,增溶剂或乳化剂,例如,有水,乙醇,异丙醇,碳酸乙酯,乙酸乙酯,苄基醇,苯甲酸苄酯,亚丙基二醇,1,3-丁基二醇油,甘油脂肪族酯,聚乙烯二醇或山梨糖醇酐的脂肪酸酯。
在本发明的化妆品的剂型是悬浮液时,作为载体成分,可利用水,乙醇或亚丙基二醇这样的液态的稀释剂,乙氧基化异硬脂基醇,聚氧乙烯山梨糖醇酯及聚氧乙烯山梨糖醇酐酯这样的悬浮剂,微晶纤维素,甲基氢氧化铝,膨润土,琼脂或黄蓍胶等。
在本发明的化妆品的剂型是含有界面-活性剂的清洁剂时,作为载体成分,可利用脂肪族醇硫酸盐,脂肪族醇醚硫酸盐,磺基琥珀酸单酯,羟乙基磺酸盐,咪唑啉鎓衍生物,甲基牛磺酸盐,肌氨酸盐,脂肪酸酰胺醚硫酸盐,烷基酰胺基甜菜碱,脂肪族醇,脂肪酸甘油酯,脂肪酸二乙醇酰胺,植物性油,羊毛脂衍生物或乙氧基化甘油脂肪酸酯等。
本发明的化妆品组合物中所包含的成分除了作为有效成分的肽和载体成分之外,还可包含化妆品组合物中常规利用的成分,例如,可包含抗氧化剂,稳定化剂,增溶剂,维生素,颜料及香料这样的常规的佐剂。
本发明的术语“皮肤状态改善”被解释为可包括使由皮肤的内在的因素或外因性因素诱发的皮肤的损伤治疗,减轻,缓和的过程或其效果等,例如,呈现在皮肤发生炎症的缓和,改善等的效果,但不限于此。
本发明的再别的实施方式涉及由二噁英类似物质诱发的疾病的预防或治疗方法。
所述预防或治疗方法可包括使受试者与包含选自由SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽的1种以上的肽的药学组合物接触的阶段。
所述药学组合物与如上说明的相同。
本发明的再别的实施方式涉及由SEQ ID NO:1、2或3的氨基酸序列组成的肽用于由二噁英类似物质诱发的疾病的预防或治疗用途。
所述由SEQ ID NO:1、2或3的氨基酸序列组成的肽与如上说明的相同。
本说明书的术语“肽”是指氨基酸残基由肽键相互结合而形成的线形的分子。本发明的肽可根据本领域公知的化学合成方法,尤其是,固态合成技术(solid-phasesynthesis techniques;Merrifield,J.Amer.Chem.Soc.85:2149-54(1963);Stewart,etal.,Solid Phase Peptide Synthesis,2nd.ed.,Pierce Chem.Co.:Rockford,111(1984))或液态合成技术(US注册专利第5,516,891号)制造。
本说明书的术语“稳定性”不仅表示体内稳定性,也表示储藏稳定性(例如,常温储藏稳定性)。
本说明书中的术语“药剂学有效量”是指对于达成上述的肽的功效或活性而言充分的量。
【发明的效果】
本发明涉及具有抵抗环境污染物质的细胞保护功效的肽及其用途,确认由SEQ IDNO:1、2或3的氨基酸序列组成的肽与被知为二噁英系列中毒性最强的2,3,7,8-四氯二苯并二噁英(以下称为TCDD)直接结合(binding)而防止皮肤内渗透,发挥防止由TCDD及微细尘内包含的多环芳族烃导致的AhR活性机理的作用,这样的直接的抵抗环境污染物质的细胞保护效果与间接阻碍与物质接触机会或通过肠壁强化而使毒性减小的以往的方法具有差别性。
【附图简述】
图1a是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽对TCDD的结合力的图。
图1b是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽对TCDD的结合力的图。
图1c是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽对TCDD的结合力的图。
图2a是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽的AhR核转位检查结果的图。
图2b是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽的AhR核转位检查结果的图。
图2c是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽的AhR核转位检查结果的图。
图2d是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽的AhR核转位检查结果的图。
图2e是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽的AhR核转位检查结果的图。
图2f是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽的AhR核转位检查结果的图。
图3a是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽的TCDDICC结果的图。
图3b是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽的TCDDICC结果的图。
图3c是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽的TCDDICC结果的图。
图4a是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽的细胞内ROS分析结果的图。
图4b是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽的细胞内ROS分析结果的图。
图4c是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽的细胞内ROS分析结果的图。
图5a是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽的CYP1A1及炎症分子RT-PCR结果的图。
图5b是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽的CYP1A1及炎症分子RT-PCR结果的图。
图5c是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽的CYP1A1及炎症分子RT-PCR结果的图。
图6a是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽的AhR核转位检查结果的图。
图6b是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽的AhR核转位检查结果的图。
图6c是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽的AhR核转位检查结果的图。
图7a是显示根据本发明的实施例的由SEQ ID NO:1的氨基酸序列组成的肽的CYP1A1及炎症分子RT-PCR结果的图。
图7b是显示根据本发明的实施例的由SEQ ID NO:2的氨基酸序列组成的肽的CYP1A1及炎症分子RT-PCR结果的图。
图7c是显示根据本发明的实施例的由SEQ ID NO:3的氨基酸序列组成的肽的CYP1A1及炎症分子RT-PCR结果的图。
【用于实施发明的最佳的形态】
涉及由SEQ ID NO:1、2或3的氨基酸序列组成的肽。
【用于实施发明的形态】
接下来,通过以下实施例更详细说明本发明。但是,这些实施例仅旨在例示本发明,本发明的范围不限于这些实施例。
【制造例:序列的合成】
将氯三苯甲基氯树脂(Chloro trityl chloride resin;CTC树脂,Nova biochemCat No.01-64-0021)70g放入反应容器,添加二氯甲烷(MC)490ml而搅拌3分钟。然后,除去溶液,放入二甲基仿酰胺(DMF)490ml而搅拌3分钟后再次除去溶剂。然后,向反应基放入700ml的二氯甲烷溶液,放入Fmoc-Tyr(tBu)-OH(Bachem,Swiss)200mmol及二异丙基乙基胺(DIEA)400mmol后搅拌而充分溶解,搅拌1小时而使反应。然后,洗涤后使甲醇和DIEA(2:1)溶解于DCM(dechloromethane)而反应10分钟,用过量的DCM/DMF(1:1)洗涤。然后,除去溶液,放入490ml二甲基仿酰胺(DMF)而搅拌3分钟后,再次除去溶剂。将脱保护溶液(20%的哌啶(Piperidine)/DMF)700ml放入反应容器,在常温搅拌10分钟后除去溶液。放入等量的脱保护溶液而再次维持10分钟反应后,除去溶液,用DMF洗涤2次,用MC洗涤1次,用DMF洗涤1次,每次各自3分钟而制造Tyr(tBu)-CTL树脂(Resin)。
向新反应器放入700ml的DMF溶液,放入Fmoc-Arg(Pbf)-OH(Bachem,Swiss)200mmol,HoBt 200mmol及HBTu 200mmol后搅拌而充分溶解。将400mmol DIEA作为级分经2次放入反应基后,最小搅拌5分钟至全部固体溶解。将溶解的氨基酸混合溶液放入包含脱保护的树脂的反应容器,在常温搅拌1小时而使反应。除去反应液,用DMF溶液搅拌3次每次5分钟后除去。取少量反应树脂,利用茚三酮测试(Nihydrin Test)检查反应程度。用脱保护溶液与上述相同地进行2次脱保护反应而制造Arg(Pbf)-Tyr(tBu)-CTL树脂。用DMF和MC充分洗涤后再次施行1次茚三酮测试后,与上述相同地施行接下来的氨基酸附着实验。依据选定的氨基酸序列而使以Fmoc-Gly-OH,Fmoc-Gly-OH,Fmoc-Gly-OH,Fmoc-Trp-OH,Fmoc-Lys(Boc)-OH顺序进行链式反应。将Fmoc-保护基用脱保护溶液反应2次,每次10分钟后充分洗涤而除去。将肽基树脂各自用DMF,MC及甲醇洗涤3次,缓慢流通氮气而干燥后,在P2O5下真空减压而完全干燥后,放入1,900ml离去溶液[三氟化醋酸(Trifluroacetic acid)81.5%,蒸馏水5.0%,茴香硫醚(Thioanisole)5.0%,苯酚(Phenol)5.0%,乙烷二硫醇(EDT,Ethanedithiol)2.5%,三异丙基硅烷(TIS,Triisopropylsilane)1.0%]后,在常温下一边摇晃一边维持2小时反应。对树脂进行过滤,将树脂用少量的TFA溶液洗涤后与母液合并。向合并的母液2,090ml添加冷醚而诱导沉淀后,离心集沉淀,用冷醚再洗涤2次。除去母液后在氮气氛下充分干燥而纯化前合成79.8g由SEQ ID NO:1组成的肽(收率:97.0%)。利用分子量测定器进行分子量测定而可得到分子量822.9(理论值:822.9)。
由SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽也以如上所述的方法进行合成。
【表1】
【实施例1:试管内结合分析】
在酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)用板(plate)上将SEQ ID NO:1,2,3肽与包被缓冲液(20mM磷酸钠pH 9.6)混合至各自成1.8mM的浓度,在4℃条件下培养一晚。然后,用含有Tween-20的磷酸盐缓冲盐水(Phosphatebuffered saline with Tween-20,PBST)洗涤后,用3%牛血清白蛋白(bovine serumalbumin,BSA)于室温阻断(blocking)2小时。然后,再次用PBST洗涤后,每孔(well)添加2μM的2,3,7,8-四氯二苯并-p-二噁英(Tetrachlorodibenzo-p-dioxin;以下称为TCDD),于室温培养2小时。然后,用PBST洗涤后,将结合有异硫氰酸荧光素(Fluoresceinisothiocyanate,FITC)的抗-TCDD抗体以抗体:PBST=1:100的比率进行处理后于室温培养2小时。然后,用PBST洗涤后用荧光测定器测定激发488nm/发射520nm值,其结果示于图1a~图1c及表2。
【表2】
序列编号 | 对照组 | 50μM | 500μM | 1000μM | 2000μM |
1 | 100% | 193% | 360% | 394% | 575% |
2 | 100% | 128% | 264% | 358% | 405% |
3 | 100% | 159% | 253% | 400% | 420% |
如可在图1a~图1c及表2确认,确认由SEQ ID NO:1、2或3的氨基酸序列组成的肽呈现对TCDD的直接的结合力。
【实施例2:AhR核转位检查(nuclear translocation test)】
将作为人角质形成细胞的HaCaT以3×105个细胞/孔的密度接种到6-孔板后培养一晚。然后,将10nM的TCDD和50μM由SEQ ID NO:1、2或3的氨基酸序列组成的肽添加到培养基而使反应30分钟后对细胞进行1小时处理,回收细胞而各自分离核和细胞质蛋白。施行利用芳烃受体(Aryl hydrocarbon reCeptor,AhR)抗体(santacruz biotechnology,USA)的蛋白印迹确认活化的AhR核移动,其结果示于图2a~图2f及表3。
【表3】
如可在图2a~图2f及表3确认,由SEQ ID NO:1、2或3的氨基酸序列组成的肽抑制由TCDD导致的AhR核移动。
【实施例3:TCDD ICC】
将作为人角质形成细胞的HaCaT以3×105个细胞/孔的密度接种到6-孔板后培养一晚。然后,将50nM的TCDD和50μM由SEQ ID NO:1、2或3的氨基酸序列组成的肽添加到培养基而使反应30分钟后,对细胞进行5分钟处理,用4%多聚甲醛(paraformaldehyde)固定30分钟。然后,洗涤3次后,在0.5%Triton X-100中反应15分钟后洗涤3次。然后,在3%BSA中阻断1小时,将结合有异硫氰酸荧光素(Fluorescein isothiocyanate,FITC)的针对TCDD的第一抗体(1:100)于4℃反应一晚。进行4,6-二脒基-2-苯基吲哚(4,6-diamidino-2-phenylindole,DAPI)染色及封固而用荧光显微镜进行观察,其结果示于图3a~图3c。
如可在图3a~图3c确认,由SEQ ID NO:1、2或3的氨基酸序列组成的肽阻碍TCDD的细胞内流入。
【实施例4:细胞内ROS分析】
将作为人角质形成细胞的HaCaT以3×105个细胞/孔的密度接种到6-孔板后培养一晚。然后,将10nM的TCDD和50μM由SEQ ID NO:1、2或3的氨基酸序列组成的肽添加到培养基而使反应30分钟后,对细胞进行24小时处理后用DCFH-DA追加处理30分钟。然后,细胞回收后,通过FACS分析对平均FL1值的变化进行观察,其结果示于图4a~图4c。
如可在图4a~图4c确认,由SEQ ID NO:1、2或3的氨基酸序列组成的肽降低因TCDD而增加的细胞内ROS水平。
【实施例5:CYP1A1及炎症分子RT-PCR】
将作为人角质形成细胞的HaCaT以3×105个细胞/孔的密度接种到6-孔板后培养一晚。然后,将10nM的TCDD和50μM由SEQ ID NO:1、2或3的氨基酸序列组成的肽添加到培养基而使反应30分钟后,对细胞进行6小时或24小时处理,回收细胞而分离RNA。RNA定量后,利用cDNA合成试剂盒(Intron,韩国)合成cDNA,利用PCR预混剂(Intron,韩国)及下示表4的各自对于CYP1A1,TNF-a,IL-6,IL-1b,COX-2的引物进行PCR。然后,在5%琼脂糖凝胶上泳动而在各样品处理条件下对上述生长因子的mRNA表达程度进行比较,其结果示于图5a~图5c。
【表4】
序列编号 | 引物 | 序列(5′-3′) |
4 | CYP1A1_F | GGATCTTTCTCTGTACCCTGG |
5 | CYP1A1_R | AGCATGTCCTTCAGCCCAGA |
6 | TNF-a_F | CGTCAGCCGATTRTGCTATCT |
7 | TNF-a_R | CGGACTCCGCAAAGTCTAAG |
8 | IL-6_F | AAAGAGGCACTGCCAGAAAA |
9 | IL-6_R | ATCTGAGGTGCCCATGCTAC |
10 | IL-1b_F | TTCGACACATGGGATAACGA |
11 | IL-1b_R | TCTTTCAACACGCAGGACAG |
12 | COX-2_F | ATCATTCACCAGGCAAATTGC |
13 | COX-2_R | GGCTTCAGCATAAAGCGTTTG |
如可在图5a~图5c确认,田SEQ ID NO:1、2或3的氨基酸序列组成的肽显示抑制因TCDD而诱导的CYP1A1及诸炎症关联因子的表达的效果。
【实施例6:AhR核转位检查】
将作为人角质形成细胞的HaCaT以3×105个细胞/孔的密度接种到6-孔板后培养一晚。将10nM的Urban particulate matters(PM,Sigma Aldrich,USA)和50μM由SEQ IDNO:1、2或3的氨基酸序列组成的肽添加到培养基而使反应30分钟后,对细胞进行1小时处理后回收细胞而各自分离核和细胞质蛋白。然后,施行利用芳烃受体(Aryl hydrocarbonreceptor,AhR)抗体(santacruz biotechnology,USA)的蛋白印迹而确认活化的AhR核移动,其结果示于图6a~图6c。
如可在图6a~图6c确认,由SEQ ID NO:1、2或3的氨基酸序列组成的肽抑制由微细尘导致的AhR核移动。
【实施例7:CYP1A1及炎症分子RT-PCR】
将作为人角质形成细胞的HaCaT以3×105个细胞/孔的密度接种到6-孔板后培养一晚。将10nM的PM(Particulate matter)和50μM由SEQ ID NO:1、2或3的氨基酸序列组成的肽添加到培养基而反应30分钟后,对细胞进行6小时或24小时处理后回收细胞而分离RNA。然后,对RNA进行定量,利用cDNA合成试剂盒(Intron,韩国)合成cDNA,利用PCR预混剂(Intron,韩国)及下示表5的各自对于CYP1A1,TNF-a,IL-6,IL-1b,COX-2的引物进行PCR。然后,在5%琼脂糖凝胶上泳动而在各样品处理条件下对上述生长因子的mRNA表达程度进行比较,其结果示于图7a~图7c。
【表5】
序列编号 | 引物 | 序列(5′-3′) |
4 | CYP1A1_F | GGATCTTTCTCTGTACCCTGG |
5 | CYP1A1_R | AGCATGTCCTTCAGCCCAGA |
6 | TNF-a_F | CGTCAGCCGATTRTGCTATCT |
7 | TNF-a_R | CGGACTCCGCAAAGTCTAAG |
8 | IL-6_F | AAAGAGGCACTGCCAGAAAA |
9 | IL-6_R | ATCTGAGGTGCCCATGCTAC |
10 | IL-1b_F | TTCGACACATGGGATAACGA |
11 | IL-1b_R | TCTTTCAACACGCAGGACAG |
12 | COX-2_F | ATCATTCACCAGGCAAATTGC |
13 | COX-2_R | GGCTTCAGCATAAAGCGTTTG |
如可在图7a~图7c确认,由SEQ ID NO:1、2或3的氨基酸序列组成的肽显示抑制因微细尘而诱导的CYP1A1及诸炎症关联因子的表达的效果。
【产业上利用可能性】
本发明涉及具有抵抗环境污染物质的细胞保护功效的肽及其用途。
序列表
<110> CAREGEN CO., LTD.
<120> 具有抵抗环境污染物质的细胞保护效果的肽及其用途
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Claims (5)
1.肽,其由SEQ ID NO:1、2或3的氨基酸序列组成。
2.由二噁英类似物质诱发的疾病的预防或治疗用药物组合物,其包含:
选自由SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽的1种以上的肽作为有效成分,及
药剂学容许的载体,
其中所述由二噁英类似物质诱发的疾病选自:氯性痤疮,***数减小,睾丸癌,***癌,子宫内膜增殖症,乳腺癌,肝毒性,免疫功能低下,高脂血症,尿道下裂,隐睾症,畸形儿出生,血管损伤,肝细胞性癌,肝的肥大,腺纤维症,体重损失,毛发损失,口腔浮肿,眼睑浮肿及胃粘膜性溃疡,且
其中所述二噁英类似物质是选自下列的1种以上:多氯化二苯并-p-二噁英(PCDD),多氯化二苯并呋喃(PCDF)及多氯化联苯(PCB)。
3.选自由SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽的1种以上的肽在制造由二噁英类似物质诱发的疾病的预防或治疗用药物组合物中的用途,
其中所述由二噁英类似物质诱发的疾病选自:氯性痤疮,***数减小,睾丸癌,***癌,子宫内膜增殖症,乳腺癌,肝毒性,免疫功能低下,高脂血症,尿道下裂,隐睾症,畸形儿出生,血管损伤,肝细胞性癌,肝的肥大,腺纤维症,体重损失,毛发损失,口腔浮肿,眼睑浮肿及胃粘膜性溃疡,且
其中所述二噁英类似物质是选自下列的1种以上:多氯化二苯并-p-二噁英(PCDD),多氯化二苯并呋喃(PCDF)及多氯化联苯(PCB)。
4.权利要求3的用途,其中所述多氯化二苯并-p-二噁英是2,3,7,8-四氯二苯并-p-二噁英(TCDD)。
5.皮肤状态改善用化妆品组合物,其包含:
选自由SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸序列组成的肽的1种以上的肽作为有效成分,及
化妆品学容许的载体。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20030003671A (ko) * | 2001-03-24 | 2003-01-10 | 주식회사 메덱스바이오 | 정향 추출물을 유효성분으로 하는 다이옥신 유사물질에대한 길항성 조성물 |
JPWO2005035554A1 (ja) * | 2003-10-10 | 2007-09-06 | 東和科学株式会社 | ダイオキシン結合材料及びダイオキシンの検出又は定量方法 |
CN101068509A (zh) * | 2004-09-01 | 2007-11-07 | 华沙整形外科股份有限公司 | 体内递送成骨诱导性制剂的聚合物包裹物 |
CN101400699A (zh) * | 2006-01-11 | 2009-04-01 | 布里斯托尔-迈尔斯·斯奎布公司 | 人胰高血糖素样肽-1调节剂及其在治疗糖尿病和相关病症中的用途 |
CN105308064A (zh) * | 2013-05-13 | 2016-02-03 | 凯尔格恩有限公司 | 用于诱导肥大细胞-特异性凋亡的肽及其用途 |
CN106488928A (zh) * | 2014-05-13 | 2017-03-08 | 凯尔格恩有限公司 | 具有抗炎症、骨形成及生发促进活性的肽及其的用途 |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5516891A (en) | 1992-06-16 | 1996-05-14 | Kinerton, Ltd. | Liquid phase synthesis of peptides and peptide derivatives |
JP2002510213A (ja) * | 1997-05-31 | 2002-04-02 | ヘキスト・シエーリング・アグレボ・ゲゼルシヤフト・ミツト・ベシユレンクテル・ハフツング | 除草剤試験法 |
US6783953B1 (en) | 1998-12-22 | 2004-08-31 | Janssen Pharmaceutica N.V. | Vascular endothelial growth factor-X |
KR100480972B1 (ko) | 2001-03-24 | 2005-05-06 | (주)내츄럴엔도텍 | 초피나무 추출물을 유효성분으로 하는 다이옥신유사물질에 대한 길항성 조성물 |
JP2003231697A (ja) | 2002-02-06 | 2003-08-19 | Japan Science & Technology Corp | ダイオキシントラッピング能を有するタンパク質 |
US8344211B2 (en) * | 2008-08-13 | 2013-01-01 | Ceres, Inc. | Plant nucleotide sequences and corresponding polypeptides |
KR100667039B1 (ko) | 2004-10-22 | 2007-01-12 | 한국생명공학연구원 | 다이옥신 노출 여부 진단용 마커 및 이를 이용하여 다이옥신 노출을 확인하는 방법 |
US8088976B2 (en) * | 2005-02-24 | 2012-01-03 | Monsanto Technology Llc | Methods for genetic control of plant pest infestation and compositions thereof |
WO2013090186A1 (en) * | 2011-12-14 | 2013-06-20 | modeRNA Therapeutics | Modified nucleic acids, and acute care uses thereof |
US8877434B2 (en) * | 2012-04-19 | 2014-11-04 | Hsinyu Lee | Method and system of detecting dioxin-like compounds |
CA2871528A1 (en) * | 2012-04-24 | 2013-10-31 | Thrombogenics N.V. | Anti-pdgf-c antibodies |
KR101943083B1 (ko) | 2017-03-30 | 2019-01-29 | (주)케어젠 | 환경오염 물질에 대한 세포 보호 효과를 갖는 펩타이드 및 이의 용도 |
KR101943081B1 (ko) * | 2017-08-31 | 2019-01-29 | (주)케어젠 | 주름 개선 활성을 나타내는 펩타이드 및 이의 용도 |
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Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20030003671A (ko) * | 2001-03-24 | 2003-01-10 | 주식회사 메덱스바이오 | 정향 추출물을 유효성분으로 하는 다이옥신 유사물질에대한 길항성 조성물 |
JPWO2005035554A1 (ja) * | 2003-10-10 | 2007-09-06 | 東和科学株式会社 | ダイオキシン結合材料及びダイオキシンの検出又は定量方法 |
CN101068509A (zh) * | 2004-09-01 | 2007-11-07 | 华沙整形外科股份有限公司 | 体内递送成骨诱导性制剂的聚合物包裹物 |
CN101400699A (zh) * | 2006-01-11 | 2009-04-01 | 布里斯托尔-迈尔斯·斯奎布公司 | 人胰高血糖素样肽-1调节剂及其在治疗糖尿病和相关病症中的用途 |
CN105308064A (zh) * | 2013-05-13 | 2016-02-03 | 凯尔格恩有限公司 | 用于诱导肥大细胞-特异性凋亡的肽及其用途 |
CN106488928A (zh) * | 2014-05-13 | 2017-03-08 | 凯尔格恩有限公司 | 具有抗炎症、骨形成及生发促进活性的肽及其的用途 |
Non-Patent Citations (2)
Title |
---|
Methods for Determination of Dioxins and Dioxin-like Compounds – A Brief Review of Recent Advances;RadivojPetronijevic等;《Procedia Food Science》;20151231;第5卷;第227-230页 * |
二噁英慢性低剂量暴露对雌性SD大鼠血清蛋白的影响;陈曦等;《中国工业医学杂志》;20091231(第3期);第163-167页 * |
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JP6896094B2 (ja) | 2021-06-30 |
EP3611182B1 (en) | 2024-02-14 |
EP3611182A1 (en) | 2020-02-19 |
CN110461864A (zh) | 2019-11-15 |
US11859016B2 (en) | 2024-01-02 |
BR112019020063A2 (pt) | 2020-04-28 |
US11597748B2 (en) | 2023-03-07 |
EA201992304A1 (ru) | 2020-02-25 |
KR101943083B1 (ko) | 2019-01-29 |
WO2018182172A1 (ko) | 2018-10-04 |
KR20180112140A (ko) | 2018-10-12 |
ES2975420T3 (es) | 2024-07-05 |
JP2020515547A (ja) | 2020-05-28 |
US20200377550A1 (en) | 2020-12-03 |
US20210371464A1 (en) | 2021-12-02 |
US11104704B2 (en) | 2021-08-31 |
EP3611182A4 (en) | 2020-10-28 |
US20230227501A1 (en) | 2023-07-20 |
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