EGFR (IC50 = 5 nM); EGFR^L858R; EGFR^L858R/T790M; EGFR^T790M; EGFR^L861Q

体外活性：埃克替尼以剂量依赖性方式抑制EGFR活性，IC50值为5 nM，在62.5 nM时完全抑制。埃克替尼选择性地单独抑制EGFR成员，包括野生型和突变型，抑制效率为61-99%。埃克替尼以剂量依赖性方式阻断人表皮样癌 A431 细胞中 EGFR 介导的细胞内酪氨酸磷酸化。同时，在我们对A431、BGC-823、A549、H460、HCT8、KB和Bel-7402细胞系进行的增殖测定中，我们发现细胞系对埃克替尼的相对敏感性为A431 > BGC-823 > A549 > H460 >知识库 > HCT8 和 Bel-7402。埃克替尼表现出广泛的抗肿瘤活性，尤其对表达较高水平 EGFR 的肿瘤特别有效。激酶测定：在体外激酶测定中，2.4 ng/μL EGFR 蛋白与 32 ng/μL Crk 在含有 1 μM 冷 ATP 和 1 μCi 32P-γ-ATP 的 25 μL 激酶反应缓冲液中混合。将混合物与 0、0.5、2.5、12.5 或 62.5 nM 埃克替尼在冰上孵育 10 分钟，然后在 30 °C 下孵育 20 分钟。在 100 °C 下用 SDS 样品缓冲液淬灭 4 分钟后，通过在 10% SDS-PAGE 凝胶中电泳来解析蛋白质混合物。然后将干燥的凝胶暴露于 PhosphorImager 以检测放射性。定量由 ImageQuant 软件执行。在该方法中，放射性信号与激酶活性成反比。细胞测定：将细胞（103 个/孔，A431、BGC-823、A549、H460、HCT8、KB 和 Bel-7402 细胞）接种到 96 孔板的含有 10% FBS 的 RPMI-1640 培养基中，并在 5 % CO2 培养箱，37°C。 24小时后，用0、0.78、1.56、3.125、6.25、12.5或25μM埃克替尼处理细胞96小时。通过从第 4 天的平均吸光度值减去第 0 天的平均吸光度值来计算细胞增殖。

体内：体内研究表明，埃克替尼在携带各种人类肿瘤衍生异种移植物的裸鼠体内表现出强烈的剂量依赖性抗肿瘤作用。该药物在高达120mg/kg/天的剂量下对小鼠具有良好的耐受性，在治疗过程中没有死亡或显著的体重减轻。最近完成了一项使用吉非替尼作为晚期癌症（NSCLC）患者主动对照的头对头随机双盲III期试验（试验注册ID:NCT01040780）。

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埃克替尼在不同类型的异种移植物中显示出抗肿瘤作用。埃克替尼在 120 mg/kg 的剂量下，对 A431、A549 和 H460 异种移植物的肿瘤生长抑制率分别为 51.5%、31.0% 和 67.4%。

将 2.4 ng/μL EGFR 蛋白和 32 ng/μL Crk 混合在 25 μL 激酶反应缓冲液中，其中含有 1 μM 冷 ATP 和 1 μCi³²P-γ-ATP，用于体外激酶测定。使用 10 分钟的冰基孵育时间在 0、0.5、2.5、12.5 或 62.5 nM 埃克替尼下孵育混合物。然后添加 20 分钟的固化时间。在 100°C 下用 SDS 样品缓冲液淬灭 4 分钟后，使用 10% SDS-PAGE 凝胶进行电泳以解析蛋白质混合物。为了检测放射性，随后将干燥的凝胶暴露。软件处理量化[1]。

在含有 10% FBS 的 RPMI-1640 培养基中，将每孔 1000 个细胞接种到 96 孔板中。然后细胞在 37°C、5% CO₂ 的培养箱中生长。一天后，将埃克替尼以 0、0.78、1.56、3.125、6.25、12.5 或 25 μM 添加到细胞中，持续一整天。细胞增殖的计算包括从第4天的平均吸光度值减去第0天的平均吸光度值[1]。

Mice: In mice with A431, A549, H460, and HCT8 tumor xenografts, the effects of three doses of icotinib (30, 60, and 120 mg/kg/dose p.o. qd) on antitumor activity and survival are assessed. In these studies, a positive control group is given 30 mg/kg/dose intraperitoneally once a week[1].

Absorption, Distribution and Excretion
Bioavailability = 52%
>90% via faeces, 9% via urine
the volume of distribution was calculated as Vz/F = 115.00 ± 63.26 l
the clearance was calculated as CL/F = 13.30 ± 4.78 l/h

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Metabolism / Metabolites
Hepatic (mainly CYP3A4, less CYP1A2)

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Biological Half-Life
5.5 hrs

Protein Binding
Icotinib binds to Sudlow's site I in subdomain IIA of Human Serum Albumin (HSA) molecule, resulting in the formation of icotinib-HSA complexes.

[1]. Icotinib (BPI-2009H), a novel EGFR tyrosine kinase inhibitor, displays potent efficacy in preclinical studies. Lung Cancer. 2012 May;76(2):177-82.

Icotinib is a potent and specific epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) Icotinib was approved in China by the SFDA in June, 2011 and in January 2014, Beta Pharma, Inc. was given a “May Proceed” from the US FDA to conduct a Phase I study for the evaluation of icotinib as a treatment of EGFR+ Non-Small Cell Lung Cancer (NSCLC).
Icotinib is an orally available quinazoline-based inhibitor of epidermal growth factor receptor (EGFR), with potential antineoplastic activity. Icotinib selectively inhibits the wild-type and several mutated forms of EGFR tyrosine kinase. This may lead to an inhibition of EGFR-mediated signal transduction and may inhibit cancer cell proliferation. EGFR, a receptor tyrosine kinase, has been upregulated in a variety of cancer cell types.

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Drug Indication
Icotinib hydrochloride is a novel epidermal growth factor receptor (EGFR)–tyrosine kinase inhibitor, exhibits encouraging efficacy and tolerability in patients with advanced non-small-cell lung cancer (NSCLC) who failed previous chemotherapy.

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Mechanism of Action
Icotinib is a highly selective, first generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) which binds reversibly to the ATP binding site of the EGFR protein, preventing completion of the signal transduction cascade. EGFR is an oncogenic receptor and patients with activating somatic mutations, such as an exon 19 deletion or exon 21 L858R mutation, within the tyrosine kinase domain display unchecked cell proliferation.

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Icotinib hydrochloride is a member of quinazolines.
Icotinib Hydrochloride is the hydrochloride salt form of icotinib, an orally available quinazoline-based inhibitor of epidermal growth factor receptor (EGFR), with potential antineoplastic activity. Icotinib selectively inhibits the wild-type and several mutated forms of EGFR tyrosine kinase. This may lead to an inhibition of EGFR-mediated signal transduction and may inhibit cancer cell proliferation. EGFR, a receptor tyrosine kinase, has been upregulated in a variety of cancer cell types.

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Icotinib, one of the leading compounds selected from our compound library, was found to be a potent and specific epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) with an IC(50) of 5 nM. When profiled with 88 kinases, Icotinib only showed meaningful inhibitory activity to EGFR and its mutants. Icotinib blocked EGFR-mediated intracellular tyrosine phosphorylation (IC(50)=45 nM) in the human epidermoid carcinoma A431 cell line and inhibits tumor cell proliferation. In vivo studies demonstrated that Icotinib exhibited potent dose-dependent antitumor effects in nude mice carrying a variety of human tumor-derived xenografts. The drug was well tolerated at doses up to 120 mg/kg/day in mice without mortality or significant body weight loss during the treatment. A head to head randomized, double blind phase III trial using Gefitinib as an active control for patients with advanced non-small cell lung cancer (NSCLC) was finished recently (Trial registration ID: NCT01040780). The data shows that Icotinib was non-inferior to Gefitinib in terms of median progression free survival (PFS) and safety superior favor to Icotinib compared to Gefitinib.[1]

C22H21N3O4

391.42

391.153

C, 67.51; H, 5.41; N, 10.74; O, 16.35

610798-31-7

Icotinib Hydrochloride;1204313-51-8;Icotinib-d4;1567366-82-8

22024915

White to off-white solid powder

1.3±0.1 g/cm3

581.3±50.0 °C at 760 mmHg

305.3±30.1 °C

0.0±1.6 mmHg at 25°C

1.649

1.88

74.73

1

7

3

29

553

0

O1C([H])([H])C([H])([H])OC([H])([H])C([H])([H])OC([H])([H])C([H])([H])OC2C([H])=C3C(C(=NC([H])=N3)N([H])C3=C([H])C([H])=C([H])C(C#C[H])=C3[H])=C([H])C1=2

QQLKULDARVNMAL-UHFFFAOYSA-N

InChI=1S/C22H21N3O4/c1-2-16-4-3-5-17(12-16)25-22-18-13-20-21(14-19(18)23-15-24-22)29-11-9-27-7-6-26-8-10-28-20/h1,3-5,12-15H,6-11H2,(H,23,24,25)

N-(3-ethynylphenyl)-2,5,8,11-tetraoxa-15,17-diazatricyclo[10.8.0.014,19]icosa-1(12),13,15,17,19-pentaen-18-amine

BPI-2009; BPI2009; BPI 2009; BPI2009H; BPI-2009H; 610798-31-7; Conmana; BPI-2009; N-(3-ethynylphenyl)-7,8,10,11,13,14-hexahydro-[1,4,7,10]tetraoxacyclododecino[2,3-g]quinazolin-4-amine; UNII-9G6U5L461Q; 9G6U5L461Q; ICOTINIB [WHO-DD]; BPI 2009H; Icotinib; Trade name: Conmana

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.39 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (6.39 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 3 中的溶解度: 0.5% CMC: 30mg/mL

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。