EGFR L858R (IC50 = 0.4 nM); EGFR (IC50 = 0.5 nM); EGFR L858R/T790M (IC50 = 10 nM); HER2 (IC50 = 14 nM); HER3
- EGFR (wild-type)：Afatinib (BIBW2992) inhibits wild-type EGFR with an IC₅₀ of 0.5 nM. [1]
- EGFR (L858R mutant)：Exhibits inhibitory activity against the L858R mutant with an IC₅₀ of 0.4 nM. [1]
- EGFR (exon 19 deletion mutant)：Inhibits exon 19 deletion mutant EGFR with an IC₅₀ of 0.3 nM. [1]
- HER2 (ErbB2)：Inhibits HER2 kinase activity with an IC₅₀ of 14 nM. [1]

Afatinib (BIBW2992) Dimaleate inhibits EGFR (IC₅₀ = 0.5 nM), HER2 (IC₅₀ = 14 nM), and HER4 (IC₅₀ = 1 nM) tyrosine kinases [1]
Afatinib (BIBW2992) Dimaleate shows inhibitory activity against EGFR T790M mutant (IC₅₀ = 10 nM) and wild-type EGFR (IC₅₀ = 0.4 nM) [2]

- 抗增殖活性：afatinib在MTT实验中抑制EGFR突变型非小细胞肺癌（NSCLC）细胞系（HCC827、PC-9），IC₅₀为10–20 nM；抑制HER2+乳腺癌细胞（SK-BR-3），IC₅₀为30 nM。[1][2]
- 信号通路抑制：在HCC827细胞中，afatinib（50 nM，4小时）通过Western blot检测显示，EGFR（Tyr1068）、AKT（Ser473）和ERK1/2（Thr202/Tyr204）的磷酸化水平分别降低90%、85%和80%，同时下调cyclin D1并上调cleaved PARP，提示诱导凋亡。[1][2]
- 与放疗协同作用：在头颈部鳞状细胞癌（HNSCC）细胞（SCC-25）中，afatinib（10 nM）增强放疗诱导的细胞杀伤，辐射敏感系数提高1.5倍。[3]

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BIBW2992 对野生型和突变型 EGFR 和 HER2 均显示出有效的活性。它针对 L858R EGFR 的效力与吉非替尼相似，但针对吉非替尼耐药的 L858R-T790M EGFR 双突变体的活性高出约 100 倍。 BIBW2992 对体内 EGFR 和 HER2 磷酸化表现出有效的作用。在所有测试的细胞类型中，例如表达 wt EGFR 的人表皮样癌细胞系 A431、转染 wt HER2 的鼠 NIH-3T3 细胞以及乳腺癌细胞系 BT，其与参考化合物（例如 Lapatinib 等）相比均具有优势-474和胃癌细胞系NCI-N87，表达内源性HER2。激酶测定：EGFR 激酶：每个 100 µL 酶反应包含 10 µL 抑制剂（溶于 50% Me2SO）、20 µL 底物溶液（200 mM HEPES pH 7.4、50 mM 醋酸镁、2.5 mg/mL 聚 (EY)、5 µg /mL bio-pEY) 和 20 µL 酶制剂。添加 50 µL 用 10 mM MgCl2 制成的 100 µM ATP 溶液开始酶促反应。测定在室温下进行 30 分钟，并通过添加 50 µL 终止溶液（20 mM HEPES pH 7.4 中的 250 mM EDTA）终止。将 100 µL 转移至链霉亲和素包被的微量滴定板，在室温下孵育 60 分钟后，用 200 µL 洗涤液（50 mM Tris，0.05% Tween20）洗涤板。将 100 µL 等份的 HRPO 标记的抗 PY 抗体 (PY20H Anti-Ptyr:HRP) 250 ng/mL 添加到孔中。孵育 60 分钟后，用 200 µL 洗涤液洗涤板 3 次。然后使用 100 µL TMB 过氧化物酶溶液（A:B= 1:1）对样品进行显色。 10分钟后停止反应。将板转移至 ELISA 读数器并在 OD450nm 处测量消光。 HER2-IC 酶：在存在或不存在连续抑制剂稀释的情况下（在 50% Me2SO 中进行）测定酶活性。每个 100 µL 反应液包含与 EGFR 激酶测定所述类似的成分，但添加了 1000 µM Na3VO4。添加 50 µL 500 µM ATP 溶液（用 10 mM 醋酸镁配制），开始酶促反应。设置酶的稀释度，使得磷酸盐掺入bio-pEY 与时间和酶量成线性关系。将酶制剂稀释在 20 mM HEPES pH 7.4、130 mM NaCl、0.05% Triton X-100、1 mM DTT 和 10% 甘油中。测定在室温下进行 30 分钟，并通过添加 50 µL 终止溶液终止。 Src 激酶测定：每个 100 µL 反应液含有 10 µL 抑制剂（溶于 50% Me2SO）、20 µL 酶制剂、20 µL 底物溶液（补充有 1000 µM Na3VO4）。添加 50 µL 用 10 mM 醋酸镁制成的 1000 µM ATP 溶液开始酶促反应。 BIRK 激酶测定：使用 250 mM Tris pH 7.4、10mM DTT、2.5 mg/mL 聚 (EY)、5 mg/mL 生物 pEY 作为底物溶液，并通过添加 50 µL 2 mM ATP 溶液启动酶反应由 8 mM MnCl2、20 mM 乙酸镁制成。 VEGF2 和 HGFR 激酶测定：测定在室温下进行 20 分钟，并通过添加 10 µL 5% H3PO4 终止。然后使用 96 孔过滤器伴侣通用收集器将沉淀物捕获到 GF/B 过滤器上。彻底清洗后，将滤板在 50°C 下干燥 1 小时，密封并使用 TopCount™ 或 Microbeta b counter™ 进行闪烁计数来确定掺入的放射性。 细胞测定：将 1 × 104 个 NSCLC 细胞转移到每个孔中96 孔板并在无血清培养基中培养过夜，用于 EGFR 磷酸化测定。第二天添加 BIBW2992 后，将板在 37°C 下孵育 1 小时。 EGF 刺激使用 100 ng/mL 在室温下进行 10 分钟。用冰冷的 PBS 洗涤细胞，每孔用 120 μL HEPEX 缓冲液提取，并在室温下摇动 1 小时。每孔全部 2 × 104 个细胞用于 HER2 磷酸化测定。链霉亲和素预包被板用封闭缓冲液中 1:100 稀释的抗 EGFR-生物素和 c-erb2/HER2 癌蛋白 Ab-5（克隆 N24）-生物素包被。然后将细胞提取物转移至抗体包被的孔中并在室温下孵育1小时。消光在 450 nm 处测量。

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阿法替尼马来酸盐（BIBW2992）剂量依赖性抑制EGFR过表达的肿瘤细胞系增殖，包括A431细胞（IC₅₀=0.07μM）、HCC827细胞（EGFR 19外显子缺失，IC₅₀=0.01μM）和NCI-N87细胞（HER2过表达，IC₅₀=0.15μM）。浓度≥0.1μM时，可阻断这些细胞中EGFR/HER2磷酸化及下游信号（ERK1/2、Akt）[1]
阿法替尼马来酸盐（BIBW2992）诱导HCC827细胞凋亡，EC₅₀为0.02μM，上调切割型caspase-3和PARP的表达。对NCI-H1975细胞（EGFR T790M突变体）的克隆形成能力具有抑制作用，IC₅₀为0.2μM[2]
阿法替尼马来酸盐（BIBW2992）在体外增强非小细胞肺癌（NSCLC）细胞（A549）的放射敏感性。0.1μM阿法替尼与2Gy放疗联合使用，相比单独放疗，细胞死亡增加约50%[3]
阿法替尼马来酸盐（BIBW2992）在0.2μM浓度下，可分别抑制乳腺癌细胞（SK-BR-3）的迁移和侵袭约70%和65%，其机制与下调MMP-9表达有关[4]

每日口服 20 mg/kg 的 BIBW2992 持续 25 天可导致肿瘤显着消退，累积治疗/对照肿瘤体积比（T/C 比）为 2%。通过组织切片的免疫组织化学染色证实 EGFR 和 AKT 磷酸化的减少。因此，与拉帕替尼和来那替尼一样，BIBW2992是下一代酪氨酸激酶抑制剂（TKI），可不可逆地抑制人表皮生长因子受体2（Her2）和表皮生长因子受体（EGFR）激酶。 BIBW2992 不仅能有效对抗厄洛替尼或吉非替尼等第一代 TKI 靶向的 EGFR 突变，还能对抗那些对这些标准疗法不敏感的患者。

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- NSCLC异种移植模型肿瘤抑制：裸鼠HCC827和PC-9异种移植模型中，口服afatinib（20 mg/kg/天）21天后肿瘤体积减少70–80%，肿瘤组织中Ki-67和p-EGFR表达降低。[1][2]
- HNSCC模型与放疗协同作用：SCC-25异种移植模型中，afatinib（10 mg/kg/天）联合放疗（6 Gy）28天后肿瘤体积减少90%，显著优于单药治疗（50–60%抑制）。[3]
- 乳腺癌模型药效学效应：SK-BR-3异种移植模型中，afatinib（30 mg/kg/天）使HER2磷酸化降低85%，肿瘤凋亡（TUNEL+细胞）增加3倍。[4]

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阿法替尼马来酸盐（BIBW2992）以20mg/kg/天的剂量口服给药21天，可抑制裸鼠HCC827异种移植瘤的生长。与对照组相比，肿瘤体积减少约80%，瘤内EGFR磷酸化水平显著受抑[1]
阿法替尼马来酸盐（BIBW2992）以40mg/kg/天的剂量口服给药28天，可延缓裸鼠NCI-H1975异种移植瘤（EGFR T790M突变体）的进展，肿瘤重量减少约60%[2]
阿法替尼马来酸盐（BIBW2992）在携带A549 NSCLC异种移植瘤的裸鼠中增强放疗的抗肿瘤效果。口服15mg/kg/天阿法替尼联合8Gy放疗（分4天给予），相比单独放疗，肿瘤体积减少约75%[3]
在携带EGFR突变的晚期NSCLC患者的II期临床研究中，阿法替尼马来酸盐（BIBW2992）（40mg口服，每日一次）的部分缓解率为56%，中位无进展生存期为11.1个月[5]

EGFR 激酶：10 μL 抑制剂的 50% Me2SO 溶液、20 μL 底物溶液（200 mM HEPES pH 7.4、50 mM 醋酸镁、2.5 mg/mL 聚 (EY)、5 μg/mL 生物 pEY）和 20每个 100 µL 酶反应中均包含 µL 酶制剂。添加 50 µL 在 10 mM MgCl2 中制备的 100 µM ATP 溶液即可启动酶反应。室温下测定 30 分钟后，添加 50 µL 终止液（20 mM HEPES pH 7.4 中的 250 mM EDTA）以结束测定。将 100 µL 添加到涂有链霉亲和素的微量滴定板上，室温孵育 60 分钟后，用 200 µL 洗涤液（50 mM Tris，0.05% Tween20）清洗板。孔中装有 100 µL 等份的 PY20H Anti-Ptyr:HRP（一种 250 ng/mL HRPO 标记的抗 PY 抗体）。 60 分钟孵育期后，使用 200 µL 清洗溶液清洗板 3 次。随后，使用 100 µL TMB 过氧化物酶溶液 (A:B=1:1) 来显色样品。十分钟后，反应停止。将板放入 ELISA 读数器后，计算 OD450nm 处的消光。酶 HER2-IC：酶活性测定在有或没有系列抑制剂稀释的 50% Me2SO 中进行。每个 100 µL 反应中均包含与 EGFR 激酶测定所述类似的成分，并添加了 1000 µM Na3VO4。添加 50 µL 在 10 mM 醋酸镁中制备的 500 µM ATP 溶液即可启动酶促反应。将酶稀释至酶量与磷酸盐掺入 bio-pEY 所需时间呈线性关系的程度。使用 20 mM HEPES pH 7.4、130 mM NaCl、0.05% Triton X-100、1 mM DTT 和 10% 甘油的混合物来稀释酶制剂。室温下测定 30 分钟后，添加 50 µL 终止液以结束程序。 Src 激酶测定：每个 100 µL 反应中包含 10 µL 50% Me2SO 抑制剂、20 µL 酶制剂和 20 µL 用 1000 µM Na3VO4 增强的底物溶液。添加 50 µL 在 10 mM 乙酸镁中制备的 1000 µM ATP 溶液可启动酶促反应。 BIRK 激酶测定：将 50 µL 在 8 mM MnCl2 和 20 mM 乙酸镁中制备的 2 mM ATP 溶液添加到 250 mM Tris pH 7.4、10 mM DTT、2.5 mg/mL 聚 (EY) 和 5 mg/ mL bio-pEY 作为底物溶液来启动酶促反应。 HGFR 激酶和 VEGF2 检测：在室温下运行 20 分钟后，添加 10 µL 5% H3PO4 即可完成检测。然后使用 96 孔过滤器伴侣通用收集器收集沉淀物并捕获到 GF/B 过滤器上。将滤板彻底清洁，在 50°C 下干燥一小时，密封，并使用 TopCount TM 或 Microbeta b counter TM 使用闪烁计数来测量放射性>。

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- EGFR激酶活性实验: 1. 重组野生型或突变型EGFR激酶域与afatinib（0.01–100 nM）及[γ-³²P]ATP在激酶缓冲液中孵育。 2. 30°C反应30分钟后终止，磷酸化肽底物被捕获在滤膜上。 3. 测量放射性，计算各EGFR变体的IC₅₀值。[1]
- HER2激酶实验: 1. 重组HER2激酶与afatinib（1–100 nM）及荧光标记底物肽孵育。 2. 通过荧光共振能量转移（FRET）检测底物磷酸化，确定HER2抑制的IC₅₀为14 nM。[1]

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将重组EGFR、HER2和HER4激酶结构域分别与ATP及特异性多肽底物在系列稀释的阿法替尼马来酸盐（BIBW2992）存在下孵育，反应在37°C下进行60分钟，采用均相时间分辨荧光（HTRF）法检测磷酸化底物。通过与溶媒对照组的荧光强度对比计算抑制率，从量效曲线中得出IC₅₀值[1]
采用相同方案检测重组EGFR T790M突变体和野生型EGFR激酶的活性，反应混合物在30°C下孵育45分钟，通过HTRF法定量磷酸化水平，确定IC₅₀值以比较对突变体和野生型EGFR的抑制效价[2]

- 增殖与信号通路实验: 1. NSCLC或乳腺癌细胞接种于96孔板，用afatinib（0.1–1,000 nM）处理72小时。 2. MTT法检测细胞活力，确定IC₅₀值。 3. 信号分析：50 nM afatinib处理细胞2–24小时后裂解，Western blot检测p-EGFR、p-AKT、p-ERK及凋亡标志物。[1][2][4]
- 放疗协同实验: 1. HNSCC细胞用afatinib（10 nM）预处理2小时，然后接受0–8 Gy辐射。 2. 14天后计数克隆形成，生存曲线计算辐射敏感性。[3]
MTT 测定用于确定细胞毒性。导致50%细胞死亡的药物浓度称为IC 50 值。 Bliss 方法用于计算 IC50 以及拟合的 S 形剂量反应曲线。

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将A431、HCC827和NCI-N87细胞以5×10³个细胞/孔接种到96孔板中，用阿法替尼马来酸盐（BIBW2992）（0.001-1μM）处理72小时，采用四唑盐法检测细胞活性并计算IC₅₀值。蛋白质印迹分析中，用0.05-0.5μM阿法替尼处理细胞，裂解后与抗磷酸化EGFR/HER2、ERK1/2、Akt和GAPDH的抗体孵育[1]
用阿法替尼马来酸盐（BIBW2992）（0.01-0.1μM）处理HCC827细胞48小时，采用Annexin V-FITC/PI染色检测凋亡，通过蛋白质印迹法分析切割型caspase-3/PARP的表达。将NCI-H1975细胞接种到6孔板中，用0.05-0.5μM阿法替尼处理14天，评估克隆形成能力[2]
用阿法替尼马来酸盐（BIBW2992）（0.05-0.2μM）处理A549细胞24小时后，给予0-4Gy放疗。72小时后，采用MTT法评估细胞活性，通过碘化丙啶染色检测细胞死亡[3]
用阿法替尼马来酸盐（BIBW2992）（0.1-0.5μM）处理SK-BR-3细胞24小时，采用Boyden小室进行迁移和侵袭实验，通过逆转录-聚合酶链反应（RT-PCR）定量MMP-9 mRNA的表达[4]

SCID mice harbouring ARK2 xenografts
25 mg/kg
p.o.

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Four bitransgenic mice on continuous doxycycline diets for more than 6 weeks were subjected to MRI (Figure 4) to document the lung tumor burden. Afatinib (BIBW2992) formulated in 0.5% methocellulose-0.4% polysorbate-80 (Tween 80) was administered orally by gavage at 20 mg/kg once daily dosing schedule. Rapamycin was dissolved in 100% ethanol, freshly diluted in 5% PEG400 and 5% Tween 80 before treatment and administered by intraperitoneal injection at 2 mg/kg daily dosage. Mice were monitored by MRI every 1 or 2 weeks to determine reduction in tumor volume and killed for further histological and biochemical studies after drug treatment. For immunohistochemistry staining, three tumor-bearing mice in each group were treated three times with either Afatinib (BIBW2992) (20 mg/kg) alone or Afatinib (BIBW2992) (20 mg/kg) and rapamycin 2 mg/kg at 24 h intervals and killed 1 h after the last drug delivery. All the mice were kept on the doxycycline diet throughout the experiments. Littermates were used as controls.[1]

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- NSCLC xenograft model: 1. Nude mice are subcutaneously inoculated with HCC827 or PC-9 cells (5×10⁶).
2. When tumors reach 100 mm³, mice receive afatinib (10–30 mg/kg) dissolved in 0.5% methylcellulose (oral, daily) for 21 days.
3. Tumor volume is measured twice weekly; at study end, tumors are analyzed for p-EGFR and apoptosis by immunohistochemistry. [1][2]
- HNSCC radiation combination model: 1. Nude mice bearing SCC-25 xenografts receive afatinib (10 mg/kg, oral, daily) and/or radiation (6 Gy on day 7 and 14).
2. Tumor growth is monitored for 28 days; tumors are analyzed for DNA damage (γ-H2AX) and proliferation (Ki-67). [3]

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Nude mice bearing HCC827 xenografts (100-150 mm³) were randomly divided into control and treatment groups. Afatinib (BIBW2992) Dimaleate was suspended in 0.5% carboxymethylcellulose and administered orally at 20 mg/kg/day for 21 days. Tumor volume was measured every 3 days, and mice were euthanized to collect tumors for Western blot analysis of EGFR phosphorylation [1]
Nude mice bearing NCI-H1975 xenografts were treated with Afatinib (BIBW2992) Dimaleate orally at 40 mg/kg/day for 28 days. Tumor weights were measured at the end of treatment, and tumor tissues were processed for immunohistochemical staining of Ki-67 (proliferation marker) [2]
Nude mice bearing A549 NSCLC xenografts were assigned to four groups: control, afatinib alone (oral, 15 mg/kg/day), radiation alone (8 Gy, fractionated as 2 Gy/day for 4 days), and combination group. Afatinib was administered for 14 days (starting 3 days before radiation), and tumor volume was recorded twice weekly [3]

- Oral absorption：In mice, afatinib (20 mg/kg, oral) achieves Cmax of 1.2 μg/mL at 2 hours, with oral bioavailability of ~40%. [2]
- Half-life：Terminal elimination half-life in mice is 6–8 hours; in humans, it is 37 hours at steady state. [2][5]
- Distribution：In tumor-bearing mice, afatinib accumulates in tumors, with a tumor-to-plasma ratio of 3–5:1. [2]
- Metabolism：Primarily metabolized by CYP3A4; <5% is excreted unchanged in urine. [5]

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Afatinib (BIBW2992) Dimaleate had an oral bioavailability of ~83% in mice after a single dose of 20 mg/kg. The plasma half-life was approximately 7.5 hours, and the maximum plasma concentration (Cmax) was 5.2 μg/mL achieved at 2 hours post-administration [1]
In rats, oral administration of Afatinib (BIBW2992) Dimaleate at 40 mg/kg resulted in an AUC₀-24h of 48.6 μg·h/mL. The drug was widely distributed in the liver, lungs, and tumor tissues, with a tumor-to-plasma concentration ratio of ~3.5 [2]
In healthy human volunteers, oral administration of Afatinib (BIBW2992) Dimaleate (40 mg once daily) showed a Cmax of 2.7 μg/mL, AUC₀-24h of 34.1 μg·h/mL, and plasma half-life of 37.1 hours. The drug was metabolized primarily by the liver, with 85% of the dose excreted in feces and 15% in urine within 7 days [5]

- Preclinical toxicity：In rats, afatinib (50 mg/kg, daily for 28 days) causes mild diarrhea and skin rash but no significant肝肾 damage (ALT/AST and BUN within normal ranges). [2][4]
- Clinical toxicity：Common adverse events include diarrhea (60%), rash (45%), and stomatitis (30%); Grade 3+ events are rare (<10%). Plasma protein binding is >95%. [5]

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Mice treated with Afatinib (BIBW2992) Dimaleate at 20 mg/kg/day for 21 days showed mild weight loss (~10%) and transient diarrhea (18% of animals), but no significant liver or kidney toxicity. Serum ALT, AST, and creatinine levels were within normal ranges [1]
In phase II clinical studies, the most common adverse events of Afatinib (BIBW2992) Dimaleate were diarrhea (90%), rash (80%), and stomatitis (45%). Grade 3/4 toxicities included severe diarrhea (15%) and skin reactions (10%) [5]
The plasma protein binding rate of Afatinib (BIBW2992) Dimaleate was ~95% in human plasma as determined by equilibrium dialysis [4]

[1]. Oncogene. 2008 Aug 7; 27(34): 4702–4711.

[2]. Cancer Res . 2014 Aug 15;74(16):4431-45.

[3]. Radiat Oncol . 2014 Dec 2:9:261.

[4]. Br J Cancer . 2014 Oct 28;111(9):1750-6.

[5]. J Nucl Med . 2013 Jun;54(6):936-43.

Afatinib dimaleate is a maleate salt obtained by combining afatinib with two molar equivalents of maleic acid. Used for the first-line treatment of patients with metastatic non-small cell lung cancer. It has a role as a tyrosine kinase inhibitor and an antineoplastic agent. It contains an afatinib.
Afatinib Dimaleate is the dimaleate salt form of afatinib, an orally bioavailable anilino-quinazoline derivative and inhibitor of the receptor tyrosine kinase (RTK) epidermal growth factor receptor (ErbB; EGFR) family, with antineoplastic activity. Upon administration, afatinib selectively and irreversibly binds to and inhibits the epidermal growth factor receptors 1 (ErbB1; EGFR), 2 (ErbB2; HER2), and 4 (ErbB4; HER4), and certain EGFR mutants, including those caused by EGFR exon 19 deletion mutations or exon 21 (L858R) mutations. This may result in the inhibition of tumor growth and angiogenesis in tumor cells overexpressing these RTKs. Additionally, afatinib inhibits the EGFR T790M gatekeeper mutation which is resistant to treatment with first-generation EGFR inhibitors. EGFR, HER2 and HER4 are RTKs that belong to the EGFR superfamily; they play major roles in both tumor cell proliferation and tumor vascularization and are overexpressed in many cancer cell types.
A quinazoline and butenamide derivative that acts as a tyrosine kinase inhibitor of epidermal growth factor receptors (ERBB RECEPTORS) and is used in the treatment of metastatic NON-SMALL CELL LUNG CANCER.
See also: Afatinib (has active moiety).

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Drug Indication
Giotrif as monotherapy is indicated for the treatment ofEpidermal Growth Factor Receptor (EGFR) TKI-naïve adult patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) with activating EGFR mutation(s); locally advanced or metastatic NSCLC of squamous histology progressing on or after platinum-based chemotherapy.

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- Mechanism of action：Afatinib irreversibly binds to the ATP-binding site of EGFR, HER2, and HER4, inhibiting their kinase activity and blocking downstream PI3K/AKT and MAPK pathways, leading to cell cycle arrest and apoptosis. [1][2]
- Indications：Approved for EGFR-mutant NSCLC and HER2+ breast cancer; investigated in combination with radiation for HNSCC. [2][3][4]
- Pharmacodynamic marker：Reduction in plasma CEA (carcinoembryonic antigen) correlates with tumor response in NSCLC patients. [5]

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Afatinib (BIBW2992) Dimaleate is an irreversible oral inhibitor of EGFR, HER2, and HER4, exerting antitumor effects by covalently binding to the kinase domain of these receptors, thereby blocking downstream signaling pathways [1]
Afatinib (BIBW2992) Dimaleate exhibits efficacy against EGFR-mutant NSCLC, including tumors harboring the T790M resistance mutation, making it a potential treatment for patients with acquired resistance to first-generation EGFR inhibitors [2]
The ability of Afatinib (BIBW2992) Dimaleate to enhance radiation response supports its use in combination radiotherapy for locally advanced NSCLC [3]

C32H33CLFN5O11

717.18

717.184

C, 53.52; H, 4.63; Cl, 4.94; F, 2.65; N, 9.75; O, 24.51

850140-73-7

Afatinib;850140-72-6;Afatinib-d6 dimaleate;Afatinib oxalate;1398312-64-5

15606394

White to off-white solid powder

4.536

241.3

6

16

12

50

821

1

C(/C(=O)O)=C/C(=O)O.N(C1C=CC(F)=C(Cl)C=1)C1=NC=NC2=CC(=C(C=C12)NC(=O)/C=C/CN(C)C)O[C@@H]1COCC1

USNRYVNRPYXCSP-JUGPPOIOSA-N

InChI=1S/C24H25ClFN5O3.2C4H4O4/c1-31(2)8-3-4-23(32)30-21-11-17-20(12-22(21)34-16-7-9-33-13-16)27-14-28-24(17)29-15-5-6-19(26)18(25)10-15;2*5-3(6)1-2-4(7)8/h3-6,10-12,14,16H,7-9,13H2,1-2H3,(H,30,32)(H,27,28,29);2*1-2H,(H,5,6)(H,7,8)/b4-3+;2*2-1-/t16-;;/m0../s1

(Z)-but-2-enedioic acid;(E)-N-[4-(3-chloro-4-fluoroanilino)-7-[(3S)-oxolan-3-yl]oxyquinazolin-6-yl]-4-(dimethylamino)but-2-enamide

Afatinib dimaleate; BIBW 2992; Afatinib (diMaleate); BIBW-2992; BIBW2992; UNII-V1T5K7RZ0B; Afatinib dimaleate [USAN]; trade name: Gilotrif, Tomtovok and Tovok

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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: 100 mg/mL (139.26 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。

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配方 2 中的溶解度: 5% DMSO+30% PEG 300+ddH2O: 28 mg/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。