CN114652717B - 盐酸萘甲唑啉的药物用途 - Google Patents
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- CNIIGCLFLJGOGP-UHFFFAOYSA-N 2-(1-naphthalenylmethyl)-4,5-dihydro-1H-imidazole Chemical compound C=1C=CC2=CC=CC=C2C=1CC1=NCCN1 CNIIGCLFLJGOGP-UHFFFAOYSA-N 0.000 claims abstract description 30
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Abstract
本发明公开了萘甲唑啉的一种药物用途。所述用途为萘甲唑啉或其药学上可接受的盐在制备促进心肌细胞增殖药物中的应用。本发明通过实验证明，萘甲唑啉可以促进大鼠心肌细胞增殖。因此，萘甲唑啉可以用于制备促进心肌细胞增殖药物以及治疗或预防心脏病的药物等相关领域，为治疗或预防心脏病例如心肌梗死提供一种新的药物和治疗思路。
Description
技术领域
本发明属于医药领域，具体涉及一种盐酸萘甲唑啉的药物用途。
背景技术
当前心血管疾病已成为威胁人类健康的头号杀手，仅心力衰竭患者全球就有4000万，已经成为人类死亡的主要原因。研究发现哺乳动物成年后心肌细胞逐渐失去增殖能力，一旦发生心梗，心肌细胞的损失将不可逆转。成年人约有20-40亿个心肌细胞，心梗发生后数小时内会造成约25％的心肌细胞损失，剩余的心肌细胞增殖能力非常有限，不足以恢复心脏的收缩功能，患者最终心衰死亡。为解决这一难题，除外科手术外，基础转化研究主要包括寻找心脏干细胞或前体细胞，通过诱导分化移植到心梗区域，然而缺乏足够确凿的分子标记物，移植效率低限制了这一技术的使用；利用重编程技术将成纤维细胞转分化为心肌细胞，或通过促进内源心肌细胞增殖的方式以补充丢失的心肌。已有的研究表明低等的脊椎动物如斑马鱼、蝾螈等心脏具有很强的再生能力，新生乳鼠也具有一定的再生能力，主要通过损伤诱导的心肌细胞增殖来实现，而这种能力在成年之后就消失了。以往人们认为成年心脏不能再生，但是大量证据表明哺乳动物心肌细胞存在缓慢的更新，并且研究发现新生的心肌细胞来源于已有的心肌。因此，越来越多的科学家们对诱导内源性心肌细胞的增殖这一策略感兴趣。因此寻找能够诱导内源性心肌细胞增殖的药物对于人类心血管疾病的治疗具有重大意义。
盐酸萘甲唑啉(Naphazoline Hcl)是一种作用于循环系统的血管活性药物，属拟肾上腺素类药，有收缩血管的药理活性，解除鼻粘膜肿胀和充血。它是一种直接作用的α肾上腺素能显效剂，可使鼻内扩张的小动脉收缩，但对β肾上腺素能受体无何影响。局部应用后，10分钟内作用即可出现，持续2～6小时。临床用于治伤风过敏性鼻炎、炎症性鼻充血、急慢性鼻炎。但并没有盐酸萘甲唑啉在诱导内源性心肌细胞增殖方面的相关报道。
发明内容
本发明的目的是提供一种萘甲唑啉或其药学上可接受的盐的新用途。
所述萘甲唑啉，CAS No.550-99-2，结构式如式I所示：
所述药学上可接受的盐具体可为盐酸盐。
本发明所提供的萘甲唑啉或其药学上可接受的盐的新用途是其在制备促进心肌细胞增殖产品中的应用。
所述心肌细胞可为人或哺乳动物的心肌细胞。所述产品可为药品。
本发明另一个目的是提供萘甲唑啉或其药学上可接受的盐在制备预防和/或治疗心血管疾病产品中的应用。所述产品可为药品。
进一步地，在本发明的一些实施方案中，所述心血管疾病可为由心肌细胞缺失、损伤或死亡而导致的心脏病。
进一步地，在本发明的一些实施方案中，上述心脏病包括但不限于心肌梗死、心力衰竭以及其他由心肌细胞缺失、损伤或死亡导致的心肌病等。
以萘甲唑啉或其药学上可接受的盐为活性成分制备的促进心肌细胞增殖产品以及制备的预防和/或治疗心血管疾病的产品也均属于本发明的保护范围。
需要的时候，在上述药物中还可以加入一种或多种药学上可接受的载体；所述载体包括药学领域常规的稀释剂、赋形剂、填充剂、粘合剂、湿润剂、崩解剂、吸收促进剂、表面活性剂、吸附载体、润滑剂等。
上述药物可以制成注射液、片剂、粉剂、颗粒剂、胶囊、口服液、膏剂、霜剂等多种形式；上述各种剂型的药物均可以按照药学领域的常规方法制备。
上述药物可通过注射、喷射、滴鼻、滴眼、渗透、吸收、物理或化学介导的方法导入机体如肌肉、皮内、皮下、静脉、粘膜组织；或是被其他物质混合或包裹后导入机体。
本发明再一个目的是提供一种体外培养心肌细胞的方法。
本发明所提供的体外培养心肌细胞的方法，包括在含心肌细胞的培养基中加入萘甲唑啉或其药学上可接受的盐。
所述萘甲唑啉或其药学上可接受的盐在所述培养基中的终浓度为0.5-2μmol/L。所述心肌细胞可为人或哺乳动物的心肌细胞。
本发明还保护一种体外促进心肌细胞增殖的方法。
本发明所提供的体外促进心肌细胞增殖的方法，包括用所述萘甲唑啉或其药学上可接受的盐处理心肌细胞。
所述萘甲唑啉或其药学上可接受的盐在处理体系中的浓度为0.5-2μmol/L。所述心肌细胞可为人或哺乳动物的心肌细胞。
本发明通过实验证明，所述萘甲唑啉或其药学上可接受的盐可以促进大鼠心肌细胞增殖。因此，萘甲唑啉或其药学上可接受的盐可以用于制备促进心肌细胞增殖药物以及治疗或预防心脏病的药物等相关领域，为治疗或预防心脏病例如心肌梗死提供一种新的药物和治疗思路。
附图说明
图1为盐酸萘甲唑啉对新生大鼠心肌细胞增殖的促进作用图。
具体实施方式
下面结合具体实施例对本发明作进一步阐述，但本发明并不限于以下实施例。所述方法如无特别说明均为常规方法。所述原材料如无特别说明均能从公开商业途径获得。
下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
下述实施例中所使用的盐酸萘甲唑啉，为盐酸萘甲唑啉的DMSO溶液。NaphazolineHcl生产商TargetMol，货号T6645。
下述实施例中“FBS”为胎牛血清。
下述实施例中使用的cTnT-mAG-hGeminin(1/110)病毒的构建方法如下：
cTnT心肌特异性启动子(如序列表中序列1所示)和mAG-hGeminin(1/110)(GenBank:NM_015895)通过pEASY-Uni Seamless Cloning and Assembly Kit(全式金CU101-01)组装克隆到pShuttle载体(Addgene 16402)上，然后采用限制性内切酶PmeI进行酶切，收集线性化质粒；将线性化质粒和pAdEasy1 DNA(Addgene 16400)共转化BJ5183感受态，筛选重组质粒并扩增，然后用重组质粒转染293A细胞(转染前一天将7.5×105个293A细胞在60mm培养皿中铺板，用含5％胎牛血清的DMEM培养液培养，至细胞数量达到1.0-1.5×106，然后利用磷酸钙共沉淀法将重组质粒转染细胞，转染后第二天，去除含共沉淀颗粒的培养液，用PBS缓冲液清洗)，然后将细胞分装入一块6孔板中(每孔加入3ml含5％胎牛血清的DMEM培养液)，静置6小时使其贴壁；6小时后覆盖琼脂糖以供形成病毒空斑(空斑应在10-21天内形成，每4-5天或培养基变黄时追加琼脂糖/DMEM混合物)；得到初始病毒后经过2-3轮扩增，氯化铯密度梯度离心纯化腺病毒。
实施例1、盐酸萘甲唑啉(Naphazoline Hcl)促进大鼠心肌细胞增殖的体外试验
(1)SD大鼠的心肌细胞的培养
分离出生3天SD大鼠的心肌细胞进行培养，DMEM高糖培养基(Hyclone)+5％马血清(GIBCO)，在37度，5％二氧化碳培养箱培养。
(2)实验分组和处理
分离SD大鼠的心肌细胞，加入5％马血清(GIBCO)+DMEM高糖培养基培养液(Hyclone)，加阿糖胞苷(终浓度20umol/L)处理以抑制非心肌细胞的生长，细胞贴壁48h后感染cTnT-mAG-hGeminin(1/110)病毒(病毒感染的MOI值＝100)，再过24小时后换成含0.5％ FBS的DMEM培养液，分组加药处理，分组如下：
a、实验组：加盐酸萘甲唑啉处理(培养基中的终浓度为1μmol/L)24小时。
b、空白对照组：加与a实验组等量DMSO处理。
(3)试验方法
上述2个分组分别处理24h后，用hoechst荧光染料染细胞核，然后用高内涵活细胞分析仪(Molecular Device)进行拍照，并分析mAG-hGeminin(1/110)阳性心肌细胞及总细胞数。
(4)结果
试验结果如图1所示。由图1可知，盐酸萘甲唑啉处理后mAG-hGeminin(1/110)阳性心肌细胞相比于空白对照组(0.5％FBS+DMSO)增加了2.7倍。Mean±SEM；**P＜0.005。
SEQUENCE LISTING
<110> 忻佑康医药科技（南京）有限公司 南京景瑞康分子医药科技有限公司
<120>盐酸萘甲唑啉的药物新用途
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 836
<212> DNA
<213> Artificial sequence
<400> 1
tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg atcctctaga 60
actatagcta gaattcgccc ttacgggccc cccctcgagg tcgggataaa agcagtctgg 120
gctttcacat gacagcatct ggggctgcgg cagagggtcg ggtccgaagc gctgccttat 180
cagcgtcccc agccctggga ggtgacagct ggctggcttg tgtcagcccc tcgggcactc 240
acgtatctcc gtccgacggg tttaaaatag caaaactctg aggccacaca atagcttggg 300
cttatatggg ctcctgtggg ggaaggggga gcacggaggg ggccggggcc gctgctgcca 360
aaatagcagc tcacaagtgt tgcattcctc tctgggcgcc gggcacattc ctgctggctc 420
tgcccgcccc ggggtgggcg ccggggggac cttaaagcct ctgcccccca aggagccctt 480
cccagacagc cgccggcacc caccgctccg tgggacgatc cccgaagctc tagagcttta 540
ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct 600
cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa 660
gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt 720
tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc 780
actcccagtt caattacagc tcttaaggct agagtactta atacgactca ctatag 836
Claims (6)
1.一种非疾病诊断和治疗目的的体外培养心肌细胞的方法，包括在含心肌细胞的培养基中加入萘甲唑啉或其药学上可接受的盐。
2.根据权利要求1所述的方法，其特征在于：所述萘甲唑啉或其药学上可接受的盐在所述培养基中的终浓度为0.5-2μmol/L。
3.根据权利要求1或2所述的方法，其特征在于：所述心肌细胞为人或哺乳动物的心肌细胞。
4.一种非疾病诊断和治疗目的的体外促进心肌细胞增殖的方法，包括用萘甲唑啉或其药学上可接受的盐处理心肌细胞。
5.根据权利要求4所述的方法，其特征在于：所述萘甲唑啉或其药学上可接受的盐在处理体系中的浓度为0.5-2μmol/L。
6.根据权利要求4或5所述的方法，其特征在于：所述心肌细胞为人或哺乳动物的心肌细胞。
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| Yuan Huang 等.An α1A-Adrenergic–Extracellular Signal-Regulated Kinase Survival Signaling Pathway in Cardiac Myocytes.《Circulation》.2007,第115卷(第6期),第763-772页. * |
| 孟繁浩等.《药物化学》.中国医药科技出版社,2016,第150-151页. * |
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