抗肿瘤环八肽Samoamide A的固相合成及其细胞毒性
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摘要
采用Fmoc固相合成法,以2-氯三苯甲基氯(CTC)树脂为固相载体,Fmoc保护的氨基酸为原料,合成了环八肽Samoamide A,其结构经~1H NMR,~(13)C NMR和LC-MS(EI)确证。研究了反应溶剂、缩合剂、切割条件和pH对Samoamide A收率的影响。结果表明:合成Samoamide A的最佳条件为:60%DCM/DMF为溶剂,O-苯并三氮唑-N,N,N′,N′-四甲基脲四氟硼酸(TBTU)为缩合剂,TFA/EDT/PhOH/H_2O/硫茴香醚为切割试剂(80/2.5/7.5/5/5,V/V/V/V/V), pH 8.0,总收率54.5%。采用MTT法研究了Samoamide A的细胞毒性。结果表明:Samoamide A浓度为50.0μg·mL~(-1)时,4T1细胞的存活率为20.79%。
The hexapeptide Samoamide A was synthesized by Fmoc solid phase synthetic method, using 2-chlorotrityl chloride(CTC) resin as solid phase carrier and Fmoc protecting amino acid as material. The influence of coupling solvents, coupling reagents, cleavage conditions and pH on the yield of Samoamide A was investigated. The structure was confrimed by ~1H NMR, ~(13)C NMR and LC-MS(EI). The optimized protocol was ascertained: 60%DCM/DMF as the coupling solvent, TBTU as the coupling reagent, TFA/EDT/PhoH/H_2O/thioanisole=80/2.5/7.5/5/5 as the cleavage reagent and pH 8.0. The yield reached 54.5%. The cytotoxicity of the synthetic product were evaluated through MTT assays against various human cancer cell lines. the results showed that when the concentration of Samoamide A reached 50.0 μg·mL~(-1), the survival rate of 4 T1 cells was only 20.79%.
The hexapeptide Samoamide A was synthesized by Fmoc solid phase synthetic method, using 2-chlorotrityl chloride(CTC) resin as solid phase carrier and Fmoc protecting amino acid as material. The influence of coupling solvents, coupling reagents, cleavage conditions and pH on the yield of Samoamide A was investigated. The structure was confrimed by ~1H NMR, ~(13)C NMR and LC-MS(EI). The optimized protocol was ascertained: 60%DCM/DMF as the coupling solvent, TBTU as the coupling reagent, TFA/EDT/PhoH/H_2O/thioanisole=80/2.5/7.5/5/5 as the cleavage reagent and pH 8.0. The yield reached 54.5%. The cytotoxicity of the synthetic product were evaluated through MTT assays against various human cancer cell lines. the results showed that when the concentration of Samoamide A reached 50.0 μg·mL~(-1), the survival rate of 4 T1 cells was only 20.79%.
引文
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[5] MULLARD A.2015 FDA drug approvals[J].Nature Reviews Drug Discovery,2016,15(2):66-73.
[6] KLINKER K P,BORGERT S J.Beyond vancomycin:The tail of the lipoglycopeptides.[J].Clinical Therapeutics,2015,37(12):2619-2636.
[7] OBERG K E,LAMBERTS S W.Somatostatin analogues in acromegaly and neuroendocrine tumours:Past,present and future[J].Endocr Relat Cancer,2016,23(12):551-566.
[8] BRUNS C,LEWIS I,BRINER U,et al.SOM230:A novel somatostatin peptidomimetic with broad somatotropin release inhibiting factor (SRIF) receptor binding and a unique antisecretory profile[J].European Journal of Endocrinology,2002,146(5):707-716.
[9] JAIN S,ZAIN J.Romidepsin in the treatment of cutaneous T-cell lymphoma[J].J Blood Med,2011,2(1):37-47.
[10] ROTHSTEIN R D,FRIEDENBERG F K.Linaclotide:A novel compound for the treatment of irritable bowel syndrome with constipation[J].Expert Opinion on Pharmacotherapy,2013,14(15):2125-2132.
[11] YAQOOB M ,KAUSHIK T.Lessons learned from peginesatide in the treatment of anemia associated with chronic kidney disease in patients on dialysis[J].Biologics,2013,7(default):243-246.
[12] ANEIROS A,GARATEIX A.Bioactive peptides from marine sources:Pharmacological properties and isolation procedures[J].Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences,2004,803(1):41-53.
[13] GOGINENI V,HAMANN M T.Marine natural product peptides with therapeutic potential:Chemistry,biosynthesis,and pharmacology[J].Biochimica Et Biophysica Acta,2017,1862(1):66-81.
[14] NAMAN C B ,RATTAN R ,NIKOULINA S E ,et al.Integrating molecular networking and biological assays to target the isolation of a cytotoxic cyclic octapeptide,Samoamide A,from an American samoan marine cyanobacterium[J].Journal of Natural Products,2017,80(3):625-633.
[15] 马丽,纪浩杰,王莹,等.多肽BRC4的固相法合成及其与RAD51(231-260)的相互作用[J].合成化学,2017,25(2):137-139.
[16] 张盼,邹胜,黄晴菲,等.新型多肽固相合成链接子{4-[叔丁氧羰基氨基-(2′,4′-二甲氧基苄基)]苯氧基乙酸的合成[J].合成化学,2014,22(5):634-637.
[17] 王小青,高杨,尹志峰,等.抗菌肽IB-367的固相合成与抑菌活性[J].合成化学,2016,24(7):600-603.
[18] 张成刚,甘一如,黄鹤.Fmoc法固相合成胸腺五肽工艺[J].化学工业与工程,2005,22(6):407-410.
[19] 唐宏琨,孙立枢,陈五岭.Fmoc固相合成JFT的工艺研究[J].现代生物医学进展,2007,7(1):85-87.
[20] YI X,YANG K,LIANG C,et al.Imaging-guided combined photothermal and radiotherapy to treat subcutaneous and metastatic tumors using iodine-131-doped copper sulfide nanoparticles[J].Advanced Functional Materials,2015,25(29):4689-4699.
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