反相高效液相色谱法对小鼠血清中基因重组人促肾上腺皮质激素前体及其代谢物的测定
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摘要
采用反相高效液相色谱(RP-HPLC)法测定分析静脉注射基因重组人促肾上腺皮质激素(ACTH)前体蛋白(ProACTH141)后小鼠血清中ProACTH141及其代谢产物ACTH(1-39)的含量变化。结果表明,ProACTH141的半衰期(T_(1/2))为21.9 min;其代谢物ACTH(1-39)在血清中的达峰时间(T_(max))为20 min。ProACTH141和代谢产生的ACTH(1-39)的药时曲线下面积(AUC)分别为538.2和88.2 (mg·L~(-1))·min~(-1)。该方法操作简便,灵敏度高,结果准确,可用于基因重组人促肾上腺皮质激素前体蛋白的药物代谢动力学分析。
The concentrations of recombinant human adrenocorticotropic(ACTH) hormone precursor protein(ProACTH141) and its metabolite ACTH(1-39) in mouse blood serum were determined by using a reversed-phase high performance liquid chromatography(RP-HPLC). In the Rp-HPLC method Inertsil~® C18 column(4.6×150 mm, ∅5 μm) was adopted, and ACTH(1-24) was used as an internal standard. Linear gradient elution was performed by proportional mixing of 0.1% TFA buffer(mobile phase A) and acetonitrile containing 0.1% TFA(mobile phase B). HPLC condition: flow rate, 1 mL·min~(-1); detection wavelength, 280 nm; column temperature, room temperature. Pharmacokinetic analysis showed that the half-life(T_(1/2)) of ProACTH141 was 21.9 min and that the peak retention time(T_(max)) of its metabolite ACTH(1-39) in the serum was 20 min. The area under the curves of ProACTH141 and ACTH(1-39) was 538.2(mg·L~(-1))·min~(-1). and 88.2(mg·L~(-1))·min~(-1), respectively. The RP-HPLC method is easy to operate, and high in sensitivity and accuracy and can hence be used for pharmacokinetic analysis of the recombinant human adrenocorticotropic hormone precursor protein.
The concentrations of recombinant human adrenocorticotropic(ACTH) hormone precursor protein(ProACTH141) and its metabolite ACTH(1-39) in mouse blood serum were determined by using a reversed-phase high performance liquid chromatography(RP-HPLC). In the Rp-HPLC method Inertsil~® C18 column(4.6×150 mm, ∅5 μm) was adopted, and ACTH(1-24) was used as an internal standard. Linear gradient elution was performed by proportional mixing of 0.1% TFA buffer(mobile phase A) and acetonitrile containing 0.1% TFA(mobile phase B). HPLC condition: flow rate, 1 mL·min~(-1); detection wavelength, 280 nm; column temperature, room temperature. Pharmacokinetic analysis showed that the half-life(T_(1/2)) of ProACTH141 was 21.9 min and that the peak retention time(T_(max)) of its metabolite ACTH(1-39) in the serum was 20 min. The area under the curves of ProACTH141 and ACTH(1-39) was 538.2(mg·L~(-1))·min~(-1). and 88.2(mg·L~(-1))·min~(-1), respectively. The RP-HPLC method is easy to operate, and high in sensitivity and accuracy and can hence be used for pharmacokinetic analysis of the recombinant human adrenocorticotropic hormone precursor protein.
引文
[1] ANDREJ SLOMINSKY,ANDREZJ SZCZESNIEWSKI,JACOBO WORTSMAN.Liquid Chromatography-Mass Spectrometry Detection of Corticotropin-Releasing Hormone and Proopiomelanocortin-Derived Peptides in Human Skin[J].Journal of Clinical Endocrinology & Metabolism,2000,10(2):3582-3588.
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[7] MICHAEL R,PRANZATELLI,TYLER J,et al.Effect of low-dose cyclophosphamide,ACTH,and IVIG combination immunotherapy on neuroinflammation in pediatric-onset OMS:A retrospective pilot study[J].Official Journal of the European Paediatric Neurology Society,2018,22(4):589-594.
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[12] GIRPEN W H,WIEGANT V M,GREVEN H M,et al.The induction of excessive grooming in the rat by intraventricular application of peptides derived from ACTH:structure-activity studies[J].Life Sciences,1975,17(4):645-652.
[13] ITO M,AIBA H,HASHIMOTO K.et al.Low-dose ACTH therapy for West syndrome[J].Neurology,2002,58:110-114.
[14] 张为民,张利红,马广智,等.斜带石斑鱼阿片黑素促皮质素原全长 cDNA 克隆及序列分析[J].水生生物学报,2007,27(6):625-630.
[15] 丁冰,杨文玲,许华容,等.HPLC 法测定乙胺吡嗪利福异烟片中有关物质的含量[J].沈阳药科大学学报,2018,35(9):750-755.
[16] 卢金淼.ACTH对婴儿痉挛症的重要性[N].医药经济报,2015-12-23(8).
[17] 白海波,杜继曾.促肾上腺皮质激素释放激素及去甲肾上腺素对高原鼠兔体液免疫的抑制作用[J].兽类学报,1997,17 (2):131-135.
[18] CARUSO C,CARNIGLIA L,DURAND D,et al.Melanocortins:Anti-inflammatory and neuroprotective peptides[J].Neurodegeneration,2012,10:93-120.
[19] KERTESZ V,CALLIGARIS D.Feldman D R,et al.Profiling of adrenocorticotropic hormone and arginine vasopressin in human pituitary gland and tumor thin tissue sections using droplet-based liquid-microjunction surface-sampling-HPLC-ESI-MS-MS[J].Anal Bioanal Chem,2015,407:5989-5998.
[2] GARDI J ,KHALIL W K ,VECSEMYKS M.Molecular forms of alpha-melanocyte-stimulating hormone in pheochromocytoma tissue[J].Endocrine research,2000,26(1):71-79.
[3] ITO N,SUGAWARA K,BODO E.Corticotropin-releasing hormone stimulates the in situ generation of mast cells from precursors in the human hair follicle mesenchyme[J].Journal of Investigative Dermatology,2010,130:995-1004.
[4] SLOMINSKY A,WORTSMAN J.Neuroendocrinology of the Skin[J].Endocrine,2000,21(5):457-487.
[5] SWARON L.WARDLAW.Hypothalamic proopiomelanocortin processing and the regulation of energy balance[J].Eur J Pharmacol,2011,660(1):213-219.
[6] MORRIS H,SCANTLEBURY,CHUN KYOUNG-CHUI,et al.Adrenocorticotropic Hormone Protects Learning and Memory Function in Epileptic Kcna1-null mice[J].Neuroscience Letters,2017,645(3):14-18.
[7] MICHAEL R,PRANZATELLI,TYLER J,et al.Effect of low-dose cyclophosphamide,ACTH,and IVIG combination immunotherapy on neuroinflammation in pediatric-onset OMS:A retrospective pilot study[J].Official Journal of the European Paediatric Neurology Society,2018,22(4):589-594.
[8] GONG SHUAI,MIAO YI-LONG,JIAO GUANG-ZHONG,et al.Dynamics and correlation of serum Cortisol and Corticosterone under different physiological or stressful conditions in mice[J].PLoS ONE,2015,2:2-14.
[9] HUANG YONGLIN,PEI YECHUN,GUANHUAI,et al.Stability and pharmacological effects of gene-recombinant wild type and mutant human Adrenocorticotropic hormone[J].Springer Science+Business Media New York,2017,34:793-799.
[10] MORTON I K,JUDITH M H.Concise dictionary of pharmacological agents:properties and synonyms[J].Springer Science & Business Media,2018,16:35-41.
[11] YALOW R S,GLICK S M,ROTH J,et al.Radioimmunoassay of human plasma ACTH[J].The Journal of Clinical Endocrinology and Metabolism.1964,24 (11):1219-1225.
[12] GIRPEN W H,WIEGANT V M,GREVEN H M,et al.The induction of excessive grooming in the rat by intraventricular application of peptides derived from ACTH:structure-activity studies[J].Life Sciences,1975,17(4):645-652.
[13] ITO M,AIBA H,HASHIMOTO K.et al.Low-dose ACTH therapy for West syndrome[J].Neurology,2002,58:110-114.
[14] 张为民,张利红,马广智,等.斜带石斑鱼阿片黑素促皮质素原全长 cDNA 克隆及序列分析[J].水生生物学报,2007,27(6):625-630.
[15] 丁冰,杨文玲,许华容,等.HPLC 法测定乙胺吡嗪利福异烟片中有关物质的含量[J].沈阳药科大学学报,2018,35(9):750-755.
[16] 卢金淼.ACTH对婴儿痉挛症的重要性[N].医药经济报,2015-12-23(8).
[17] 白海波,杜继曾.促肾上腺皮质激素释放激素及去甲肾上腺素对高原鼠兔体液免疫的抑制作用[J].兽类学报,1997,17 (2):131-135.
[18] CARUSO C,CARNIGLIA L,DURAND D,et al.Melanocortins:Anti-inflammatory and neuroprotective peptides[J].Neurodegeneration,2012,10:93-120.
[19] KERTESZ V,CALLIGARIS D.Feldman D R,et al.Profiling of adrenocorticotropic hormone and arginine vasopressin in human pituitary gland and tumor thin tissue sections using droplet-based liquid-microjunction surface-sampling-HPLC-ESI-MS-MS[J].Anal Bioanal Chem,2015,407:5989-5998.