建立用于胰岛素类制剂药物评价的正常胰岛素-正常葡萄糖钳夹技术
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摘要
目的建立使用生长抑素的正常胰岛素-正常葡萄糖钳夹技术,为胰岛素类制剂药物研究搭建平台。方法 8名男性健康受试者参加试验,首先以150μg·h-1的速度输注生长抑素抑制内源性胰岛素,5 min后以0. 15 mU·kg~(-1)·min~(-1)的速度输注人胰岛素搭建胰岛素平台;估算并调整葡萄糖输注率,使血糖维持在目标范围(4. 5～5. 0 mmol·L~(-1))。稳态期后60 min,皮下注射赖脯胰岛素0. 2U·kg~(-1),并继续维持稳态至注射后480 min。期间观察血清胰岛素、C肽及血糖水平的变化。结果 7名健康受试者完成试验。试验开始后25～30 min进入稳态期,稳态期后30 min的C肽水平为(0. 41±0. 10)μg·L~(-1),钳夹结束时降至(0. 05±0. 04)μg·L~(-1)。稳态期间血清胰岛素水平维持在(10. 1±2. 6)mU·L~(-1)。稳态期间血糖水平(4. 67±0. 23) mmol·L~(-1),变异系数5. 1%。结论小剂量输注生长抑素可以建立稳定的正常胰岛素-正常葡萄糖钳夹平台,用于胰岛素类制剂的药物研究。
Objective To establish an euinsulinemic euglycemic clamp technique for the evaluation of insulin preparations. Methods An euinsulinemic euglycemic clamp technique was applied in 8 healthy male subjects. Firstly somatostatin was infused at a rate of 150 μg·h-1 to inhibit endogenous insulin secretion and 5 minutes later continuous insulin infusion at a rate of 0. 15 mU·kg~(-1)·min~(-1) was performed. Plasma glucose was measured in order to clamp the glucose level at 4. 5-5. 0 mmol·L~(-1) by adjusting the glucose infusion rate. 60 minutes after steady state of euglycemia clamp,insulin lispro was injected subcutaneously at 0. 2 U·kg~(-1) and the euglycemic stage was maintained for the next 480 minutes. Blood insulin,C-peptide and glucose were observed during the period. Results Seven in eight subjects successfully completed the trial. The steady stage was reached 25 to 30 minutes after the beginning of the experiment. The serum C-peptide concentrations was( 0. 41 ± 0. 10) μg·L~(-1) at the first 30 min of steady stage and decreased to( 0. 05 ± 0. 04) μg·L~(-1) at the terminal stage of clamp period. The steady-state serum insulin concentration was( 10. 1 ± 2. 6) mU·L~(-1) and glucose lever was( 4. 67 ± 0. 23) mmol·L~(-1) with the coefficient of variation of 5. 1%. Conclusion A high-quality euinsulinemic euglycemicclamp technique with continuous somatostatin infusion was successfully established for the evaluation of insulin preparations in Chinese healthy male volunteers.
Objective To establish an euinsulinemic euglycemic clamp technique for the evaluation of insulin preparations. Methods An euinsulinemic euglycemic clamp technique was applied in 8 healthy male subjects. Firstly somatostatin was infused at a rate of 150 μg·h-1 to inhibit endogenous insulin secretion and 5 minutes later continuous insulin infusion at a rate of 0. 15 mU·kg~(-1)·min~(-1) was performed. Plasma glucose was measured in order to clamp the glucose level at 4. 5-5. 0 mmol·L~(-1) by adjusting the glucose infusion rate. 60 minutes after steady state of euglycemia clamp,insulin lispro was injected subcutaneously at 0. 2 U·kg~(-1) and the euglycemic stage was maintained for the next 480 minutes. Blood insulin,C-peptide and glucose were observed during the period. Results Seven in eight subjects successfully completed the trial. The steady stage was reached 25 to 30 minutes after the beginning of the experiment. The serum C-peptide concentrations was( 0. 41 ± 0. 10) μg·L~(-1) at the first 30 min of steady stage and decreased to( 0. 05 ± 0. 04) μg·L~(-1) at the terminal stage of clamp period. The steady-state serum insulin concentration was( 10. 1 ± 2. 6) mU·L~(-1) and glucose lever was( 4. 67 ± 0. 23) mmol·L~(-1) with the coefficient of variation of 5. 1%. Conclusion A high-quality euinsulinemic euglycemicclamp technique with continuous somatostatin infusion was successfully established for the evaluation of insulin preparations in Chinese healthy male volunteers.
引文
[1]EUROPEAN MEDICINES AGENCY. Guideline on non-clinical and clinical development of similar biological medicinal products containing recombinant human insulin and insulin analogues[EB/OL].London:EMA. 2015-02-26[2018-12-19]. https://www. ema. europa. eu/documents/scientific-guideline/guidelinenon-clinical-clinical-development-similar-biological-medicinal-products-containing_en-0. pdf.
[2]谭惠文,余叶蓉,李秀钧.正常血糖-高胰岛素钳夹技术在胰岛素制剂药物代谢动力学研究中的应用进展[J].华西医学,2017,32(11):1801-1804.
[3]STROWSKI M Z,PARMAR R M,BLAKE A D,et al. Somatostatin inhibits insulin and glucagon secretion via two receptors subtypes:an in vitro study of pancreatic islets from somatostatin receptor 2 knockout mice[J]. Endocrinology,2000,141(1):111-117.
[4]DEFRONZO R A,TOBIN J D,ANDRES R. Glucose clamp technique:a method for quantifying insulin secretion and resistance[J].Am J Physiol,1979,237(3):E214-E223.
[5]DIAMOND PROJECT GROUP. Incidence and trends of childhood type 1 diabetes worldwide 1990-1999[J]. Diabet Med,2006,23(8):857-866.
[6]MICHAE D J,SOREN N,NIDHI G,et al. Insulin clearance is different in men and women[J]. Metabolism, 2012,61(4):525-530.
[7]贾伟平,陈蕾,项坤三,等.扩展高胰岛素-正葡萄糖钳夹技术的建立[J].中华内分泌代谢杂志,2001,17(5):8-11.
[8]袁涛,赵维纲,付勇,等. 40/60混合重组人胰岛素注射液的人体药代动力学和药效动力学[J].中华临床营养杂志,2016,24(1):1-7.
[2]谭惠文,余叶蓉,李秀钧.正常血糖-高胰岛素钳夹技术在胰岛素制剂药物代谢动力学研究中的应用进展[J].华西医学,2017,32(11):1801-1804.
[3]STROWSKI M Z,PARMAR R M,BLAKE A D,et al. Somatostatin inhibits insulin and glucagon secretion via two receptors subtypes:an in vitro study of pancreatic islets from somatostatin receptor 2 knockout mice[J]. Endocrinology,2000,141(1):111-117.
[4]DEFRONZO R A,TOBIN J D,ANDRES R. Glucose clamp technique:a method for quantifying insulin secretion and resistance[J].Am J Physiol,1979,237(3):E214-E223.
[5]DIAMOND PROJECT GROUP. Incidence and trends of childhood type 1 diabetes worldwide 1990-1999[J]. Diabet Med,2006,23(8):857-866.
[6]MICHAE D J,SOREN N,NIDHI G,et al. Insulin clearance is different in men and women[J]. Metabolism, 2012,61(4):525-530.
[7]贾伟平,陈蕾,项坤三,等.扩展高胰岛素-正葡萄糖钳夹技术的建立[J].中华内分泌代谢杂志,2001,17(5):8-11.
[8]袁涛,赵维纲,付勇,等. 40/60混合重组人胰岛素注射液的人体药代动力学和药效动力学[J].中华临床营养杂志,2016,24(1):1-7.