CN107987133A - 小分子多肽及其用途 - Google Patents
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- C—CHEMISTRY; METALLURGY
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Abstract
本发明公开了一种小分子多肽，小分子多肽的氨基酸序列如SEQ ID No.1所示。本发明还提供了小分子多肽的用途。本发明的多肽具有良好的抗寄生虫活性，同时具备抗菌活性，稳定性强，结构易于调控，具有很明显的应用潜力。
Description
技术领域
本发明涉及一种小分子多肽。更具体地说，本发明涉及一种具有抗寄生虫和抗菌活性的小分子多肽。
背景技术
寄生虫病作为一种常见疾病，影响了世界人民的健康。例如，疟疾在非洲、亚洲和拉丁美洲等热带和亚热带地区成为一个严重的社会问题，每年有大约两亿人感染疟疾，超过五十万人死亡。目前，有效作用于寄生虫疾病的药物不多，而耐药性却经常发生。因此，寻找新的寄生虫疾病药物极为迫切，特别地，寻找一种具有寄生虫活性同时兼具其它生物活性的药物具有重要的意义。
发明内容
本发明的一个目的是提供一种小分子多肽及其用途，该小分子多肽稳定性强，并具有具有良好的抗寄生虫和抗菌活性。
为了实现根据本发明的这些目的和其它优点，提供了一种小分子多肽，小分子多肽的氨基酸序列如SEQ ID No.1所示。
优选的是，所述的小分子多肽，所述小分子多肽的等电点为3.7。
本发明还提供了一种小分子多肽的用途，用于制备抗菌药物。
本发明还提供了一种小分子多肽的用途，用于制备抗寄生虫药物。
本发明至少包括以下有益效果：
1、本发明的多肽具有良好的抗寄生虫活性，同时具备抗菌活性。
2、本发明方法是一种小分子多肽，稳定性强，结构易于调控，具有很明显的应用潜力。
3、本发明小分子多肽与大分子多肽相比合成方法简便，易于大规模生产。
本发明的其它优点、目标和特征将部分通过下面的说明体现，部分还将通过对本发明的研究和实践而为本领域的技术人员所理解。
具体实施方式
下面结合实施例对本发明做进一步的详细说明，以令本领域技术人员参照说明书文字能够据以实施。
实施例1
一种小分子多肽，小分子多肽的氨基酸序列如SEQ ID No.1所示；
SEQ ID No.1：AEDDCQQNEMPSTYVDDRNKFYYVSTWCQGHDD。
所述小分子多肽的等电点为3.7。
所述小分子多肽的的分子量为3961.17g/mol。
所述小分子多肽的制备方法：
选用氨基酸-王树脂作为载体(树脂)，用二氯甲烷将树脂充分溶胀，用二甲基甲酰胺清洗几遍，用适当浓度的DBLK，将Fmoc-保护基团脱出，之后用二甲基甲酰胺清洗数遍，洗去DBLK，称取适合的缩合剂苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸盐和活化剂甲基吗啉以及C端第二个Fmoc-保护氨基酸(Fmoc-Pro-OH)进行偶联，通过茚三酮检测法进行检测确保连接比较完全，用二甲基甲酰胺清洗几遍，洗去残留的各种残基，活化剂和缩合剂，依氨基酸序列进行偶联，将所有的氨基酸连接结束后，脱去最后的Fmoc-保护基团，用切割液裂解，去除树脂和氨基酸保护基团，得到小分子多肽的粗品，送质谱确认产品分子量3961.17g/mol符合理论值。
实施例2
一、实验方法
在灭菌试管中加入1mL浓度为1×106个/mL的菌液，加入1mg本发明所提供的小分子多肽，37℃培养24小时后，培养基收集起来用倍比稀释，稀释倍数为10倍和涂布培养法检测活菌数。
二、实验结果：
表1 本发明所述小分子多肽的杀菌率(％)
| 细菌 | ATCC 6538 | ATCC 25922 | ATCC 10231 | ATCC 9372 |
| 杀菌率 | 99.5％ | 99.7％ | 99.7％ | 99.9％ |
由表1可知，本发明所提供的小分子多肽，对金黄色葡萄球菌(ATCC 6538)、大肠埃希氏菌(ATCC 25922)、白假丝酵母(ATCC 10231)、枯草芽孢杆菌黑色变种(ATCC 9372)都具有很强的杀菌性，其杀菌率达99％以上。可见，本发明的小分子多肽可用于制备抗菌药物。
实施例3
采用荧光细胞分类分析(FACS)定量红细胞的寄生虫培养法来评估本发明产品的抗寄生虫活性。将实施例1得到的多肽溶解在1ml DMSO中，然后用由25mmol/L 4-羟乙基哌嗪乙磺酸缓冲液，32mmol/L NaHCO3和10％人血浆配成的RPMI 1640细胞培养介质稀释200倍。样品溶液随后依次用2倍稀释的方法配成11种不同浓度，将1ml，2×106/ml的寄生虫加入溶液中，在37℃放于5％CO2培养箱中培养48h。加入1ml 3％二脒基苯基吲哚溶液，再培养18h，用荧光细胞分类分析仪测定红细胞数，可以得出不同浓度样品对寄生虫的抑制率，计算出IC50值。
表2 多肽对寄生虫的半数有效浓度(IC50)
| 寄生虫 | 疟原虫 | 毛滴虫 |
| IC50(umol/mL) | 1.3±1.2 | 0.9±1.8 |
从实施例3的结果可以看出，本发明的多肽经抗寄生虫实验表明，该化合物具有强的抗寄生虫活性。可见，本发明的小分子多肽可用于制备抗寄生虫药物。
尽管本发明的实施方案已公开如上，但其并不仅仅限于说明书和实施方式中所列运用，它完全可以被适用于各种适合本发明的领域，对于熟悉本领域的人员而言，可容易地实现另外的修改，因此在不背离权利要求及等同范围所限定的一般概念下，本发明并不限于特定的细节和这里示出与描述的实施例。
SEQUENCE LISTING
<110> 广西中医药大学
<120> 小分子多肽及其用途
<130> application
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 33
<212> PRT
<213> 多肽
<400> 1
Ala Glu Asp Asp Cys Gln Gln Asn Glu Met Pro Ser Thr Tyr Val Asp
1 5 10 15
Asp Arg Asn Lys Phe Tyr Tyr Val Ser Thr Trp Cys Gln Gly His Asp
20 25 30
Asp
Claims (4)
1.一种小分子多肽，其特征在于，小分子多肽的氨基酸序列如SEQ ID No.1所示。
2.如权利要求1所述的小分子多肽，其特征在于，所述小分子多肽的等电点为3.7。
3.一种如权利要求1所述的小分子多肽的用途，其特征在于，用于制备抗菌药物。
4.一种如权利要求1所述的小分子多肽的用途，其特征在于，用于制备抗寄生虫药物。
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