2010年美國NCI研究藥物指導(dǎo)委員會生物標(biāo)志物工作委員會（The Biomarkers Task Force of the NCI Investigational Drug Steering Committee）發(fā)布了新藥早期臨床試驗生物標(biāo)志物研究開發(fā)指南：Guidelines for the Development and Incorporation of Biomarker Studies in Early Clinical Trials of Novel Agents.

各類Biomarker定義及作用

其中pharmacodynamic（PD）提供藥物是否有作用的直接生物學(xué)證據(jù)及毒理學(xué)基礎(chǔ)信息，因而美國很多創(chuàng)新藥物臨床一期和二期中選擇合適的pharmacodynamic（PD）生物標(biāo)志物，用于藥物在人體生物學(xué)效應(yīng)監(jiān)測。

文中談到Biomarker研究和檢測的三個考量因素：

1. 標(biāo)志物科學(xué)性：Strength of science

2. 實驗穩(wěn)定性：Assay robustness

3. 實驗可行性：Assay Feasibility

案例一： 阿斯利康 AZD1208 全球多中心，開放標(biāo)簽，First-in-human

2018年5月Nature出版集團(tuán)British Journal of Cancer在線發(fā)表了，阿斯利康AZD1208，一種原發(fā)性整合型莫洛尼病毒激酶抑制劑，在血液腫瘤和實體腫瘤的全球多中心，開放標(biāo)簽，臨床一期實驗數(shù)據(jù)。本次臨床研究包括全球多個權(quán)威腫瘤中心：MD Anderson Cancer Center血液腫瘤科，日本National Cancer Center Hospital乳腺腫瘤科，美國Dana-Farber Cancer Institute 腫瘤科，英國Royal MarsdenHospital 前列腺癌靶向治療和藥物開發(fā)部，加拿大瑪格麗特公主醫(yī)院Princess Margaret Hospital 安大略癌癥研究所，華盛頓大學(xué)醫(yī)學(xué)院腫瘤中心等。

研究目的：首要目的：AZD1208在急性髓細(xì)胞白血病（AML）(ClinicalTrials.gov, NCT01489722)和advanced solid malignancies(NCT01588548)，劑量遞增研究，每日口服一次的最大耐受劑量，藥物安全性和耐藥性。次要研究目標(biāo)：藥物PK和有效性的初步證據(jù)，及反應(yīng)藥效pharmacodynamic（PD）的生物標(biāo)志物。

Study objectives

Both dose-escalation studies recruiting patients with AML (ClinicalTrials.gov, NCT01489722) and advanced solid malignancies (NCT01588548) were phase I, open-label, multicentre studies designed to identify the maximum tolerated dose (MTD) and evaluate the safety and tolerability of AZD1208 administered orally once daily (QD). Secondary objectives included evaluation of the drug PK and preliminary evidence of efficacy. The AML study also explored pharmacodynamic (PD) biomarkers.

入組病人基本資料

劑量遞增安全性分析

藥效pharmacodynamic（PD）作者選用Simple western（Nanopro1000）分析protein level changes of phosphorylated 4E-BP1 S65

案例二：KB004, a first in class monoclonal antibody targeting the receptor tyrosine kinase EphA3 一期臨床研究

  2016年Leukemia Research發(fā)表了，KaloBios PharmaceuticalsKB004，一種靶向EphA3的單克隆抗體，在惡性血液病，全球9個中心，開放標(biāo)簽，臨床一期實驗數(shù)據(jù)。本次臨床研究包括全球多個權(quán)威腫瘤中心：MD Anderson Cancer Center血液腫瘤科，邁阿密大學(xué)血液腫瘤科，斯坦福大學(xué)腫瘤所，澳大利亞阿爾弗雷德醫(yī)院和莫納什大學(xué)，澳大利亞皇家布里斯班婦女醫(yī)院，美國克利夫蘭診所，澳大利亞韋斯特米德醫(yī)院，澳大利亞皇家阿德萊德醫(yī)院，美國H.Lee Moffitt癌癥中心。

藥效pharmacodynamic（PD）作者選用Simple western（Peggy Sue，NIA）系統(tǒng)分析分選微量CD34+細(xì)胞 EphA3表達(dá)。

Pharmacodynamic (PD) assessments were performed at baseline and serially thereafter, to determine the degree of EphA3 expression on tumor cells CD34+ cells were isolated from marrow samples by immune-magnetic separation and cryopreserved at ?80C for batch analysis . Subsequently, cells were thawed, washed and immediately lysed for NIA analysis. Equal numbers of cells for each sample were loaded for analysis into the NIA instrument (PeggySue system, ProteinSimple, Santa Clara, CA). In the instrument, proteins from each sample were separated based on charge using capillary electrophoresis. EphA3 was detected using E11F12 antibod

之前，Simple Western還在多個臨床試驗中被用于監(jiān)測Biological Response。