CN112156104B - 一种治疗抑郁症的药物 - Google Patents
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Abstract
本发明公开了一种治疗抑郁症的药物,属于神经系统药物领域。本发明所述药物以DNA四面体为活性成分,加上药学上可接受的辅助性成分制备而成。本发明的药物可对抑郁症有显著的疗效,应用前景良好。
Description
技术领域
本发明属于神经系统药物领域,尤其涉及一种治疗抑郁症的药物。
背景技术
抑郁症又称抑郁障碍,以显著而持久的心境低落为主要临床特征,是心境障碍的主要类型。临床可见心境低落与其处境不相称,情绪的消沉可以从闷闷不乐到悲痛欲绝,自卑抑郁,甚至悲观厌世,可有自杀企图或行为;甚至发生木僵;部分病例有明显的焦虑和运动性激越;严重者可出现幻觉、妄想等精神病性症状。每次发作持续至少2周以上、长者甚或数年,多数病例有反复发作的倾向,每次发作大多数可以缓解,部分可有残留症状或转为慢性。
治疗上,一线用药氟西汀,起效时间长,不利于急性发作时自杀风险控制,且对1/3的患者无效。新药氯胺酮是一种快速抗抑郁药,但伴随着各种严重副作用,且仍对约30%的患者无效。由于抑郁症目前发病机理不明确,临床上尚无完全有效的治疗方法。
四面体骨架核酸(tetrahedral framework nucleic acids,TFNAs)又称DNA四面体(tetrahedral DNA,TDN)、四面体DNA纳米结构,是一种由多条(通常是4条)单链DNA通过链间碱基互补配对形成的一种四面体结构,其代谢不会产生对身体有害的物质,生物相容性高,可用作某些药物的载体,也用于合成某些用于体内的分子检测探针。
目前未见将TFNAs应用于抑郁症治疗的报道。
发明内容
本发明要解决的问题是:提供一种新的治疗抑郁症的药物。
本发明的技术方案如下:
一种治疗抑郁症的药物,所述药物以DNA四面体为活性成分,加上药学上可接受的辅助性成分制备而成。
进一步地,所述DNA四面体由4条DNA单链通过碱基互补配对合成。
进一步地,所述DNA单链的序列如SEQ ID NO.1~4所示。
进一步地,所述DNA四面体的制备方法为:
将四条DNA单链溶于缓冲液中,加热到95℃保持10min,再降温至4℃维持20min以上。
进一步地,所述缓冲液pH=8.0。
进一步地,所述缓冲液含有10mM Tris-HCl,50mM MgCl2。
进一步地,所述药物是抗焦虑的药物。
DNA四面体在制备治疗抑郁症的药物中的用途。
进一步地,述DNA四面体由4条DNA单链通过碱基互补配对合成。
进一步地,所述DNA单链的序列如SEQ ID NO.1~4所示。
本发明的药物能通过增加海马区神经元树突分支的数目和长度,增加树突上树突棘(突触)的数量,发挥良好地治疗抑郁症的作用。
显然,根据本发明的上述内容,按照本领域的普通技术知识和惯用手段,在不脱离本发明上述基本技术思想前提下,还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式,对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1:TFNAs电泳图。
图2:透射电镜观察图。
图3:动态光散射结果。
图4:行为学实验结果。EPM:高架迷宫实验;FST:强迫游泳实验;TST:尾部悬吊实验;SPT:蔗糖偏好实验。
图5:海马区神经元观察。其中第一、二行为整体观,第三行为DG区,第四行为CA3区,第五行为CA1区,第六行为树突棘观察。
具体实施方式
实施例lTFNAs的合成与鉴定
1.合成方法
将四条DNA单链(S1、S2、S3、S4)溶解于TM Buffer(10mM Tris-HCl,50mM MgCl2,pH=8.0)中,使得四条DNA单链的终浓度为1000nM,充分混匀后迅速加热至95℃保持10分钟,之后迅速降温至4℃并维持20分钟以上,即可得到四面体骨架核酸TFNAs。
四条单链的序列(5′→3′)如下:
S1:
ATTTATCACCCGCCATAGTAGACGTATCACCAGGCAGTTGAGACGAACATTCCTAAGTCTGAA(SEQID NO.1)
S2:
ACATGCGAGGGTCCAATACCGACGATTACAGCTTGCTACACGATTCAGACTTAGGAATGTTCG(SEQID NO.2)
S3:
ACTACTATGGCGGGTGATAAAACGTGTAGCAAGCTGTAATCGACGGGAAGAGCATGCCCATCC(SEQID NO.3)
S4:
ACGGTATTGGACCCTCGCATGACTCAACTGCCTGGTGATACGAGGATGGGCATGCTCTTCCCG(SEQID NO.4)
2.鉴定
由PAGE胶结果可见TFNAs大小约为200bp(图1);透射电镜可见散在的点状物体,并能观察到部分点状物体呈现四面体形态(图2);动态光散射可见,TFNAs的大小分布均匀,在10nm左右(图3),与理论值相符。
通过前述鉴定结果,可以认为TFNAs被成功合成。
以下将以实验例的形式对本发明进一步说明。
实验例1抑郁症动物模型治疗实验
1.方法
1.1模型
6-8周大的c57小鼠,饲喂皮质酮(皮质酮掺入日常饮用水,浓度:0.1mg/ml,c57小鼠日饮水量4-7ml),连续喂养3-4周,使小鼠在TST(尾部悬吊实验)和FST(强迫游泳实验)不动时间百分比相比正常小鼠显著延长。得到抑郁症模型。
1.2分组处理
将小鼠分为4组:对照组(Control,简称C),TFNA组(简称T),疾病组(Cort,简称CO),治疗组(Cort+TFNA,简称CO+T)
处理方式如下:
1)对照组:正常小鼠每次注射生理盐水100ul,隔天一次,一共注射5次。
2)TFNA组:正常小鼠每次注射TFNAs溶液(实施例1的方法制备)100ulμL,浓度1mM,隔天一次,一共注射5次。
3)疾病组:抑郁症模型小鼠每次注射生理盐水100μL,隔天一次,一共注射4次。
4)治疗组:抑郁症模型小鼠每次注射TFNAs溶液(实施例1的方法制备)100μL,浓度1mM,隔天一次,一共注射4次。
1.3行为学实验
对各组小鼠进行尾部悬吊实验(TST)、强迫游泳实验(FST)、高架迷宫实验(EPM)、蔗糖偏好实验(SPT)。每组进行5次实验,计算平均值。
1.4病理学染色
使用高尔基染色对小鼠海马区神经元进行检测。
2.结果
2.1行为学实验结果
行为学实验结果如图4所示。具体的:
啮齿动物具有探究新环境的习性,但又厌恶强光和开阔地,EPM就是利用啮齿类动物对新异环境的探究行为和对高悬着的开臂的恐惧形成的矛盾冲突状态。动物克服对开放臂的恐惧,探索新异环境的时间越长,则表明动物抑郁程度轻,反之,抑郁程度重。EPM结果显示,在开放臂探索时间上,疾病组(CO)较对照组(C)缩短,而治疗组(CO+T)较疾病组延长。该结果表明,TFNAs具有抗抑郁作用。
FST对动物提供了一个无可回避的压迫状态,动物在该环境(水中)拼命挣扎试图逃跑又无法逃脱,一段实验后,动物表现出“不动状态”,反映了动物的“行为绝望状态”,不动状态的时间(简称“不动时间”)越长,则动物越抑郁。FST结果显示,在强迫游泳的不动时间上,疾病组较对照组延长,而治疗组较疾病组缩短。该结果表明,TFNAs可以抗抑郁。
TST的原理是利用小鼠悬尾后企图逃脱但又无法逃脱,从而放弃挣扎,进入特有的抑郁不动状态,实验过程中记录动物不动时间来反映抑郁状态,不动时间越长,抑郁越严重。TST结果显示,在尾部悬吊的不动时间上,疾病组较对照组延长,而治疗组较疾病组缩短。该结果表明,TFNAs可以抗抑郁。
SPT是基于奖励的测试,快感缺乏或经历快乐的能力降低,是抑郁症的核心症状。啮齿动物出生时对甜的食物或溶液有兴趣,如果其对糖水的偏好降低,则表示动物更抑郁。SPT结果显示,在蔗糖偏好性上,疾病组的偏好性(以糖水摄入量占总流体摄入量的比例体现)相对于对照组降低,而治疗组较疾病组提高。该结果表明,TFNAs可以抗抑郁。
2.2病理学染色
海马是调节抑郁症患者情绪状态的重要脑区之一。研究表明,抑郁症动物的海马树突密度(体现为树突分支数量和长度)、树突棘密度均显著减少,表明树突、树突棘的改变在抑郁症病理过程中发挥了重要作用(Glucocorticoids and hippocampal atrophy inneuropsychiatric disorders.Archives General Psychiatry.2000:57(10),925–935;Synaptic dysfunction in depres-sion:Potential therapeutic targets.Science2012;338:68–72;抑郁症CUS模型大鼠海马CA1区及CA3区内树突棘数目减少,中国组织化学与细胞化学杂志,2016年2月第25卷1期)。
高尔基染色结果如图5所示,TFNA组比对照组、治疗组相比疾病组,其海马区神经元树突分支数量、长度显著增加,树突棘的密度也显著增加。
该结果表明:TFNAs能通过增加海马区神经元树突分支的数目和长度,增加树突上树突棘(突触)的数量,发挥治疗抑郁症的作用。
综上,TFNAs对抑郁症有显著的疗效,将其用于制备治疗抑郁症的药物,将具有良好的前景。
SEQUENCE LISTING
<110> 四川大学
<120> 一种治疗抑郁症的药物
<130> GYKH1118-2020P0111881CC
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 63
<212> DNA
<213> 人工序列(Artificial sequence)
<400> 1
atttatcacc cgccatagta gacgtatcac caggcagttg agacgaacat tcctaagtct 60
gaa 63
<210> 2
<211> 63
<212> DNA
<213> 人工序列(Artificial sequence)
<400> 2
acatgcgagg gtccaatacc gacgattaca gcttgctaca cgattcagac ttaggaatgt 60
tcg 63
<210> 3
<211> 63
<212> DNA
<213> 人工序列(Artificial sequence)
<400> 3
actactatgg cgggtgataa aacgtgtagc aagctgtaat cgacgggaag agcatgccca 60
tcc 63
<210> 4
<211> 63
<212> DNA
<213> 人工序列(Artificial sequence)
<400> 4
acggtattgg accctcgcat gactcaactg cctggtgata cgaggatggg catgctcttc 60
ccg 63
Claims (4)
1.DNA四面体作为唯一活性成分在制备治疗抑郁症的药物中的用途;所述DNA四面体由4条DNA单链通过碱基互补配对合成;所述DNA单链的序列如SEQ ID NO.1~4所示。
2.如权利要求1所述的用途,其特征在于:所述DNA四面体的制备方法为:将四条DNA单链溶于缓冲液中,加热到95℃保持10min,再降温至4℃维持20min以上。
3.如权利要求2所述的用途,其特征在于:所述缓冲液pH=8.0。
4.如权利要求2或3所述的用途,其特征在于:所述缓冲液含有10mM Tris-HCl,50mMMgCl 2。
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CN109196103A (zh) * | 2016-05-10 | 2019-01-11 | 俄亥俄州立创新基金会 | 自组装的3d rna笼形纳米粒 |
CN109806275A (zh) * | 2017-11-22 | 2019-05-28 | 四川大学 | Dna四面体在促神经修复药物制备中的用途 |
CN110257053A (zh) * | 2019-07-02 | 2019-09-20 | 上海交通大学 | 一种形状可定制的框架核酸纳米发光体及其制备方法和应用 |
CN111544597A (zh) * | 2020-05-25 | 2020-08-18 | 四川大学 | 一种抑菌作用强的dna四面体复合物 |
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CN107669697A (zh) * | 2017-11-22 | 2018-02-09 | 四川大学 | Dna四面体在抑制阿尔兹海默细胞凋亡中的应用 |
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Publication number | Priority date | Publication date | Assignee | Title |
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CN109196103A (zh) * | 2016-05-10 | 2019-01-11 | 俄亥俄州立创新基金会 | 自组装的3d rna笼形纳米粒 |
CN109806275A (zh) * | 2017-11-22 | 2019-05-28 | 四川大学 | Dna四面体在促神经修复药物制备中的用途 |
CN110257053A (zh) * | 2019-07-02 | 2019-09-20 | 上海交通大学 | 一种形状可定制的框架核酸纳米发光体及其制备方法和应用 |
CN111544597A (zh) * | 2020-05-25 | 2020-08-18 | 四川大学 | 一种抑菌作用强的dna四面体复合物 |
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