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Afatinib (formerly BIBW 2992; BIBW-2992; brand name: Gilotrif), is a potent, covalent/irreversible, and orally bioavailable dual (EGFR/ErbB) receptor tyrosine kinase (RTK) inhibitor with anticancer activity. Afatinib is an FDA-approved anticancer medication used to treat lung cancer that is not small cell (NSCLC). In the USA, Gilotrif is the brand name under which it is sold. It is 100 times more active against the Gefitinib-resistant L858R-T790M EGFR mutant. It irreversibly binds to and inhibits EGFR/HER2, including EGFR(wt), EGFR(L858R), EGFR(L858R/T790M), and HER2. In cell-free assays, its IC50 values are 0.5 nM, 0.4 nM, 10 nM, and 14 nM, respectively.
Physicochemical Properties
| Molecular Formula | C24H25CLFN5O3 |
| Molecular Weight | 485.94 |
| Exact Mass | 485.162 |
| Elemental Analysis | C, 59.32; H, 5.19; Cl, 7.30; F, 3.91; N, 14.41; O, 9.88 |
| CAS # | 850140-72-6 |
| Related CAS # | Afatinib dimaleate;850140-73-7;Afatinib-d6;1313874-96-2;Afatinib oxalate;1398312-64-5;(R)-Afatinib;439081-17-1;Afatinib-d4 |
| PubChem CID | 10184653 |
| Appearance | White to light yellow solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 676.9±55.0 °C at 760 mmHg |
| Melting Point | 100 - 102 °C |
| Flash Point | 363.2±31.5 °C |
| Vapour Pressure | 0.0±2.1 mmHg at 25°C |
| Index of Refraction | 1.668 |
| LogP | 3.59 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 34 |
| Complexity | 702 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | 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 |
| InChi Key | ULXXDDBFHOBEHA-CWDCEQMOSA-N |
| InChi Code | InChI=1S/C24H25ClFN5O3/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/h3-6,10-12,14,16H,7-9,13H2,1-2H3,(H,30,32)(H,27,28,29)/b4-3+/t16-/m0/s1 |
| Chemical Name | (E)-N-[4-(3-chloro-4-fluoroanilino)-7-[(3S)-oxolan-3-yl]oxyquinazolin-6-yl]-4-(dimethylamino)but-2-enamide |
| Synonyms | BIBW2992; Afatinib free base; BIBW 2992; BIBW 2992; Afatinib; trade name: Gilotrif, Tomtovok and Tovok |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
EGFRL858R (IC50 = 0.4 nM); EGFR (wt) (IC50 = 0.5 nM); ErbB4 (IC50 = 1 nM); EGFRL858R/T790M (IC50 = 10 nM); HER2 (IC50 = 14 nM)
- 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) inhibits EGFR (IC₅₀ = 0.5 nM), HER2 (IC₅₀ = 14 nM), and HER4 (IC₅₀ = 1 nM) tyrosine kinases [1] Afatinib (BIBW2992) shows inhibitory activity against EGFR T790M mutant (IC₅₀ = 10 nM) and wild-type EGFR (IC₅₀ = 0.4 nM) [2] |
| ln Vitro |
- Antiproliferative activity：Afatinib inhibits proliferation of EGFR-mutant non-small cell lung cancer (NSCLC) cell lines (HCC827, PC-9) with IC₅₀ values of 10–20 nM, and HER2+ breast cancer cells (SK-BR-3) with an IC₅₀ of 30 nM in MTT assays. [1][2] - Signal pathway inhibition：In HCC827 cells, afatinib (50 nM, 4 hours) reduces phosphorylation of EGFR (Tyr1068), AKT (Ser473), and ERK1/2 (Thr202/Tyr204) by 90%, 85%, and 80%, respectively, as measured by Western blot. It also downregulates cyclin D1 and upregulates cleaved PARP, indicating apoptosis induction. [1][2] - Synergy with radiation：In head and neck squamous cell carcinoma (HNSCC) cells (SCC-25), afatinib (10 nM) enhances radiation-induced cell killing, increasing the radiation sensitivity factor by 1.5-fold. [3] BIBW2992 exhibits potent inhibition of EGFR and HER2 in both wild-type and mutant forms. It has comparable potency to gefitinib against L858R EGFR, but it is approximately 100 times more active against the L858R-T790M EGFR double mutant that is resistant to gefitinib. In vivo, BIBW2992 demonstrates strong effects on the phosphorylation of both EGFR and HER2. When compared to reference compounds (like Lapatinib et al.), it performs well in all tested cell types, including human epidermoid carcinoma cell line A431 that expresses EGFR, murine NIH-3T3 cells transfected with HER2, breast cancer cell line BT-474, and gastric cancer cell line NCI-N87 that express endogenous HER2.[1] Afatinib (BIBW2992) dose-dependently inhibited the proliferation of EGFR-overexpressing tumor cell lines, including A431 (IC₅₀ = 0.07 μM), HCC827 (EGFR exon 19 deletion, IC₅₀ = 0.01 μM), and NCI-N87 (HER2-overexpressing, IC₅₀ = 0.15 μM). It blocked EGFR/HER2 phosphorylation and downstream signaling (ERK1/2, Akt) in these cells at concentrations ≥ 0.1 μM [1] Afatinib (BIBW2992) induced apoptosis in HCC827 cells with an EC₅₀ of 0.02 μM, increasing cleaved caspase-3 and PARP levels. It also suppressed clonogenicity of NCI-H1975 cells (EGFR T790M mutant) with an IC₅₀ of 0.2 μM [2] Afatinib (BIBW2992) enhanced the radiosensitivity of non-small cell lung cancer (NSCLC) cells (A549) in vitro. Combination of 0.1 μM afatinib with 2 Gy radiation increased cell death by ~50% compared to radiation alone [3] Afatinib (BIBW2992) inhibited the migration and invasion of breast cancer cells (SK-BR-3) by ~70% and ~65% at 0.2 μM, respectively, by downregulating MMP-9 expression [4] |
| ln Vivo |
Tumor growth inhibition in NSCLC xenografts：Oral afatinib (20 mg/kg, daily) reduces tumor volume by 70–80% in HCC827 and PC-9 xenografts in nude mice after 21 days, with decreased Ki-67 and p-EGFR expression in tumor tissues. [1][2] - Synergy with radiation in HNSCC models：In SCC-25 xenografts, afatinib (10 mg/kg, daily) combined with radiation (6 Gy) reduces tumor volume by 90% after 28 days, significantly more than either treatment alone (50–60% inhibition). [3] - Pharmacodynamic effects in breast cancer models：In SK-BR-3 xenografts, afatinib (30 mg/kg, daily) decreases HER2 phosphorylation by 85% and increases tumor apoptosis (TUNEL+ cells) by 3-fold. [4] Afatinib (0-20 mg/kg, Orally, daily for 25 days) exhibits a significant decrease in tumor growth and phosphorylation of AKT, HER2, EGFR, and HER3. Afatinib (15 mg/kg, Orally, in a schedule of 5 days on plus 2 days off, for two weeks) strongly inhibits the growth of HKESC-2 tumor. Afatinib (BIBW2992) inhibited tumor growth in nude mice bearing HCC827 xenografts when administered orally at 20 mg/kg/day for 21 days. Tumor volume was reduced by ~80% compared to the control group, and intratumoral EGFR phosphorylation was significantly suppressed [1] Afatinib (BIBW2992) delayed tumor progression in nude mice bearing NCI-H1975 xenografts (EGFR T790M mutant) at an oral dose of 40 mg/kg/day for 28 days, resulting in a ~60% reduction in tumor weight [2] Afatinib (BIBW2992) augmented the antitumor effect of radiation in nude mice bearing A549 NSCLC xenografts. Oral administration of 15 mg/kg/day afatinib plus 8 Gy radiation (fractionated over 4 days) reduced tumor volume by ~75% compared to radiation alone [3] In a phase II clinical study of patients with advanced NSCLC harboring EGFR mutations, Afatinib (BIBW2992) (40 mg orally once daily) showed a partial response rate of 56% and a median progression-free survival of 11.1 months [5] |
| Enzyme Assay |
- EGFR kinase activity assay:
1. Recombinant wild-type or mutant EGFR kinase domains are incubated with afatinib (0.01–100 nM) and [γ-³²P]ATP in kinase buffer. 2. After 30 minutes at 30°C, reactions are stopped, and phosphorylated peptide substrates are captured on filters. 3. Radioactivity is measured, and IC₅₀ values are calculated for each EGFR variant. [1] - HER2 kinase assay: 1. Recombinant HER2 kinase is incubated with afatinib (1–100 nM) and fluorescently labeled substrate peptide. 2. Kinase activity is measured via fluorescence resonance energy transfer (FRET) to detect substrate phosphorylation. 3. The IC₅₀ for HER2 inhibition is determined as 14 nM. [1] The human EGFR wild type and EGFR L858R/T790M double mutant tyrosine kinase domains are fused to GST and extracted. Next, enzyme activity is measured with and without the inhibitor BIBW2992, which is serially diluted in 50% DMSO. Biotinylated pEY (bio-pEY) is added as a tracer substrate and a random polymer, pEY (4:1), is used as the substrate. Utilizing the baculovirus system, the HER2 kinase domain is cloned and extracted in a manner akin to that of EGFR kinase. Supplementary information contains specifics about the assays conducted for EGFR, HER2, SRC, BIRK, and VEGFR2 kinase activity. Recombinant EGFR, HER2, and HER4 kinase domains were individually incubated with ATP and specific peptide substrates in the presence of serial dilutions of Afatinib (BIBW2992). Reactions were carried out at 37°C for 60 minutes, and phosphorylated substrates were detected using a homogeneous time-resolved fluorescence (HTRF) assay. Inhibition rates were calculated by comparing fluorescence intensity with vehicle controls, and IC₅₀ values were derived from dose-response curves [1] Recombinant EGFR T790M mutant and wild-type EGFR kinase domains were tested using the same protocol. The reaction mixture was incubated at 30°C for 45 minutes, and phosphorylation was quantified by HTRF. IC₅₀ values were determined to compare inhibitory potency against mutant and wild-type EGFR [2] |
| Cell Assay |
- Proliferation and signaling assay:
1. NSCLC or breast cancer cells are seeded in 96-well plates and treated with afatinib (0.1–1,000 nM) for 72 hours. 2. Cell viability is measured by MTT assay to determine IC₅₀ values. 3. For signaling analysis, cells are treated with 50 nM afatinib for 2–24 hours, lysed, and p-EGFR, p-AKT, p-ERK, and apoptotic markers are detected by Western blot. [1][2][4] - Radiation synergy assay: 1. HNSCC cells are pre-treated with afatinib (10 nM) for 2 hours, then irradiated with 0–8 Gy. 2. Clonogenic survival is assessed by counting colonies after 14 days; survival curves are used to calculate radiation sensitivity. [3] For the EGFR phosphorylation test, 1 × 10 4 NSCLC cells are plated into each well of a 96-well plate and grown for an entire night in serum-free medium. The following day, the plates are incubated at 37 °C for one hour following the addition of BIBW2992. EGF stimulation is carried out at room temperature for 10 minutes using 100 ng/mL. Following an hour of shaking at room temperature and an extraction using 120 μL of HEPEX buffer per well, the cells are cleaned with ice-cold PBS. HER2 phosphorylation assay uses 2 × 10 4 cells per well in total. The c-erb2/HER2 oncoprotein Ab-5(Clone N24)-Biotin and anti-EGFR-Biotin are coated on streptavidin precoated plates at a 1:100 dilution in blocking buffer. Once in the antibody-coated wells, cell extracts are allowed to sit at room temperature for one hour. Measurement of extinction occurs at 450 nm. A431, HCC827, and NCI-N87 cells were seeded in 96-well plates at 5×10³ cells/well and treated with Afatinib (BIBW2992) (0.001-1 μM) for 72 hours. Cell viability was measured using a tetrazolium-based assay to calculate IC₅₀ values. For Western blot analysis, cells were treated with 0.05-0.5 μM afatinib, lysed, and probed with antibodies against phosphorylated EGFR/HER2, ERK1/2, Akt, and GAPDH [1] HCC827 cells were treated with Afatinib (BIBW2992) (0.01-0.1 μM) for 48 hours. Apoptosis was detected by Annexin V-FITC/PI staining, and cleaved caspase-3/PARP expression was analyzed by Western blot. NCI-H1975 cells were seeded in 6-well plates and treated with 0.05-0.5 μM afatinib for 14 days to assess clonogenicity [2] A549 cells were treated with Afatinib (BIBW2992) (0.05-0.2 μM) for 24 hours, followed by radiation (0-4 Gy). After 72 hours, cell viability was assessed by MTT assay, and cell death was detected by propidium iodide staining [3] SK-BR-3 cells were treated with Afatinib (BIBW2992) (0.1-0.5 μM) for 24 hours. Migration and invasion assays were performed using Boyden chambers, and MMP-9 mRNA expression was quantified by RT-PCR [4] |
| Animal Protocol |
- 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] Athymic NMRI-nu/nu female mice[1] 20 mg/kg Oral administration 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] Nude mice bearing HCC827 xenografts (100-150 mm³) were randomly divided into control and treatment groups. Afatinib (BIBW2992) 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) 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] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Following oral administration, time to peak plasma concentration (Tmax) is 2 to 5 hours. Maximum concentration (Cmax) and area under the concentration-time curve from time zero to infinity (AUC0-∞) values increased slightly more than dose proportional in the range of 20 to 50 mg. The geometric mean relative bioavailability of 20 mg tablets was 92% as compared to an oral solution. Additionally, systemic exposure to afatinib is decreased by 50% (Cmax) and 39% (AUC0-∞), when administered with a high-fat meal compared to administration in the fasted state. Based on population pharmacokinetic data derived from clinical trials in various tumor types, an average decrease of 26% in AUCss was observed when food was consumed within 3 hours before or 1 hour after taking afatinib. In humans, excretion of afatinib is primarily via the feces. Following administration of an oral solution of 15 mg afatinib, 85.4% of the dose was recovered in the feces and 4.3% in urine. The parent compound afatinib accounted for 88% of the recovered dose. The volume of distribution of afatinib recorded in healthy male volunteers is documented as 4500 L. Such a high volume of distribution in plasma suggests a potentially high tissue distribution. The apparent total body clearance of afatinib as recorded in healthy male volunteers is documented as being a high geometric mean of 1530 mL/min. Metabolism / Metabolites Enzyme-catalyzed metabolic reactions play a negligible role for afatinib in vivo. Covalent adducts to proteins were the major circulating metabolites of afatinib. Biological Half-Life Afatinib is eliminated with an effective half-life of approximately 37 hours. Thus, steady-state plasma concentrations of afatinib were achieved within 8 days of multiple dosing of afatinib resulting in an accumulation of 2.77-fold (AUC0-∞) and 2.11-fold (Cmax). In patients treated with afatinib for more than 6 months, a terminal half-life of 344 h was estimated. - 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] Afatinib (BIBW2992) 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) 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) (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] |
| Toxicity/Toxicokinetics |
Hepatotoxicity Elevations in serum aminotransferase levels are common during afatinib therapy occurring in 20% to 50% of patients, but rising above 5 times the upper limit of the normal range in only 1% to 2%. Hepatic failure is said to have occurred in 0.2% of patients and to have resulted in several fatalities. Hepatotoxicity appears to be a class effect among protein kinase inhibitors of EGFR2, although liver injury appears to be more frequent and more severe with gefitinib than with afatinib and erlotinib. Specific details of the liver injury associated with afatinib such as latency, serum enzyme pattern, clinical features and course, have not been published. Other EGFR inhibitors, such as erlotinib and gefitinib typically cause liver injury arising within days or weeks of starting therapy and presenting abruptly with hepatocellular enzyme elevations and a moderate-to-severe course. Immunoallergic and autoimmune features are not common. The rate of clinically significant liver injury and hepatic failure is increased in patients with preexisting cirrhosis or hepatic impairment due to liver tumor burden. Likelihood score: D (possible cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation No information is available on the clinical use of afatinib during breastfeeding. Because afatinib is about 95% bound to plasma proteins, the amount in milk is likely to be low. However, its half-life is about 37 hours and it might accumulate in the infant. the manufacturer recommends that breastfeeding be discontinued during afatinib therapy and for 2 weeks after the last dose. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding In vitro binding of afatinib to human plasma proteins is approximately 95%. Afatinib binds to proteins both non-covalently (traditional protein binding) and covalently. - 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] Mice treated with Afatinib (BIBW2992) 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) 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) was ~95% in human plasma as determined by equilibrium dialysis [4] |
| References |
[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. |
| Additional Infomation |
Pharmacodynamics Aberrant ErbB signaling triggered by receptor mutations, and/or amplification, and/or receptor ligand overexpression contributes to the malignant phenotype. Mutation in EGFR defines a distinct molecular subtype of lung cancer. In non-clinical disease models with ErbB pathway deregulation, afatinib as a single agent effectively blocks ErbB receptor signaling resulting in tumor growth inhibition or tumor regression. NSCLC tumors with common activating EGFR mutations (Del 19, L858R) and several less common EGFR mutations in exon 18 (G719X) and exon 21 (L861Q) are particularly sensitive to afatinib treatment in non-clinical and clinical settings. Limited non-clinical and/or clinical activity was observed in NSCLC tumors with insertion mutations in exon 20. The acquisition of a secondary T790M mutation is a major mechanism of acquired resistance to afatinib and gene dosage of the T790M-containing allele correlates with the degree of resistance in vitro. The T790M mutation is found in approximately 50% of patients' tumors upon disease progression on afatinib, for which T790M targeted EGFR TKIs may be considered as a next line treatment option. Other potential mechanisms of resistance to afatinib have been suggested preclinically and MET gene amplification has been observed clinically. At the same time, the effect of multiple doses of afatinib (50 mg once daily) on cardiac electrophysiology and the QTc interval was evaluated in an open-label, single-arm study in patients with relapsed or refractory solid tumors. Ultimately, no large changes in the mean QTc interval (i.e., >20 ms) were detected in the study. - 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] Afatinib (BIBW2992) 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) 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) to enhance radiation response supports its use in combination radiotherapy for locally advanced NSCLC [3] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.14 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.14 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.14 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 4: 2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 10 mg/mL Solubility in Formulation 5: 5 mg/mL (10.29 mM) in 0.5% Methylcellulose/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0579 mL | 10.2893 mL | 20.5787 mL | |
| 5 mM | 0.4116 mL | 2.0579 mL | 4.1157 mL | |
| 10 mM | 0.2058 mL | 1.0289 mL | 2.0579 mL |