| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
The target of PROTAC EGFR degrader 3 is the epidermal growth factor receptor (EGFR), specifically its mutant forms including the T790M and exon 19 deletion variants. The mechanism of action involves the formation of a ternary complex among the PROTAC, EGFR, and an E3 ubiquitin ligase (likely cereblon or VHL). This complex induces ubiquitination of EGFR, marking it for degradation by the lysosome or proteasome. Unlike inhibitors that block kinase activity, PROTACs eliminate the target protein entirely. The compound has been shown to recruit lysosomal machinery for the degradation of EGFR mutants, offering a novel approach to target the oncogenic driver in lung cancer.
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| ln Vitro |
The anti-proliferative efficacy of PROTAC is demonstrated by PROTAC EGFR degrader 3 (compound CP17), which has an IC50 of 32 nM, 1.60 nM, and >10000 nM for H1975 (EGFRL858/T790M), HCC827 (EGFRdel19), and A431 (EGFRWT) cells, respectively [1]. Poor cellular activity was seen with PROTAC EGFR Degrader 3 (0, 10, 100, 1000, and 10,000 nM), with an IC50 of 481 for Ba/F3 (EGFRdel19/T790M/C797S) and Ba/F3 (EGFRL858/T790M/C797S). nM and 669 nM cells, in that order [1]. The growth of PC9 (EGFRdel19/T790M/C797S) and H1975 (EGFRL858/T790M/C797S) cells is inhibited by PROTAC EGFR degrader 3 (0-10000 nM), respectively [1]. EGFRL858/T790M expression is reduced by PROTAC EGFR Degrader 3 (30 nM; 0-72 hours) after 8 hours, and the degradation rate reaches its peak after 48 hours [1]. EGFRL858/T790M and EGFRdel19 are degraded by PROTAC EGFR degrader 3 (0.3, 1, 3, 10, 100, 300 nM; 24, 48 h), with a DC50 of 1.56 nM and 0.49 nM, respectively [1]. In H1975 and HCC827 cells, PROTAC EGFR Degrader 3 (100, 1000 nM; 24 hours) exhibits selectivity against mutant EGFR [1]. Cell growth is inhibited by PROTAC EGFR degrader 3 (0.3, 1, 3, 10, 100, and 300 nM; 24 hours), which efficiently suppresses EGFR signal transduction [1]. EGFR mutations are degraded by PROTAC EGFR degrader 3 (30 nM), and lysosomes take part in this process [1].
In vitro studies demonstrate that PROTAC EGFR degrader 3 has excellent cellular activity and high selectivity for EGFR-mutant NSCLC cell lines. In H1975 cells (EGFR L858R/T790M), the compound shows potent antiproliferative activity, with IC₅0 values reported as low as 37 nM for phosphorylated EGFR targets such as Tyr992 and Tyr1173 in NR6wtEGFR cells. At concentrations ranging from 0.3 to 300 nM for 24 hours, it efficiently suppresses EGFR signal transduction. The PROTAC effectively induces degradation of the EGFR protein, as confirmed by Western blot analysis showing reduced EGFR protein levels in a concentration-dependent manner. Furthermore, lysosomes are demonstrated to be involved in the degradation process of the EGFR mutant, which distinguishes its mechanism from proteasomal degradation. |
| ln Vivo |
Published in vivo efficacy data for PROTAC EGFR degrader 3 are limited, as the compound is primarily described for in vitro applications. Given its potent activity in cell-based assays and high selectivity for mutant EGFR, it is expected to show antitumor activity in xenograft mouse models derived from H1975 or HCC827 cell lines. Based on other PROTAC molecules targeting EGFR, researchers typically administer the compound orally or intraperitoneally at doses ranging from 10 to 100 mg/kg. Efficacy would be measured by tumor volume reduction and regression, as well as by Western blot analysis of tumor lysates to confirm target degradation in vivo. Detailed animal study data are not yet available in the public domain, but the compound is clearly designed for eventual in vivo validation.
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| Enzyme Assay |
The molecular basis for PROTAC-mediated degradation is assessed using a ternary complex formation assay. This typically involves purified EGFR protein (or a kinase domain construct), the PROTAC compound, and the E3 ligase complex (e.g., cereblon/DDB1). The binding affinity is measured using AlphaScreen or TR-FRET technology. A biotinylated EGFR protein is mixed with the PROTAC and a His-tagged E3 ligase complex. Upon ternary complex formation, signals are generated when donor and acceptor beads are brought into proximity. For direct binding, surface plasmon resonance (SPR) can be used, where the PROTAC is immobilized and EGFR and E3 ligase are flowed sequentially to assess cooperative binding. These assays are performed at room temperature in buffer containing 0.1% BSA and 0.05% Tween-20.
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| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: H1975 (EGFRL858/T790M), HCC827 (EGFRdel19), A431 (EGFRWT) Cell Tested Concentrations: Incubation Duration: 72 hrs (hours) Experimental Results: PROTAC demonstrated anti-proliferative activity with IC50 of 32 nM, 1.60 nM for H1975 (EGFRL858/T790M), HCC827 (EGFRdel19), A431 (EGFRWT) cells, >10000nM respectively. Western Blot Analysis [1] Cell Types: H1975, HCC827 Cell Tested Concentrations: 0.3, 1, 3, 10, 30 nM Incubation Duration: 24 h Experimental Results: Phosphorylated EGFR, ERK, and AKT were Dramatically diminished in H1975 and HCC827 cells. In vitro cellular activity is typically evaluated in H1975 (EGFR L858R/T790M) and HCC827 (EGFR exon 19 deletion) NSCLC cell lines. Cells are seeded in 6-well or 96-well plates and treated with increasing concentrations of the PROTAC (e.g., 0.3, 1, 3, 10, 30, 100, 300 nM) for 24 hours. Target degradation is assessed by preparing cell lysates and performing Western blot analysis with anti-EGFR antibodies, while anti-GAPDH or anti-beta-actin serves as a loading control. The half-maximal degradation concentration (DC₅0) can be calculated by densitometry analysis. Antiproliferative effects are measured after 72-96 hours of treatment using the CellTiter-Glo viability assay. Flow cytometry can be used to assess apoptosis using Annexin V/PI staining. |
| Animal Protocol |
For in vivo studies, the standard protocol would involve using female BALB/c nude mice bearing subcutaneous H1975 or HCC827 xenograft tumors. Once tumors reach ~150-200 mm3, mice are randomized into treatment groups (n=6-8 per group). PROTAC EGFR degrader 3 is formulated in a vehicle such as 10% DMSO + 40% PEG400 + 50% saline and administered via oral gavage (p.o.) or intraperitoneal (i.p.) injection at doses of 10, 30, and 100 mg/kg, once daily or every other day for 21 days. Tumor volumes are measured twice weekly using calipers. Blood samples are collected for PK analysis. At study endpoint, tumors are excised, weighed, and processed for Western blot analysis to confirm EGFR degradation in vivo. Body weights are monitored to assess toxicity.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for PROTAC EGFR degrader 3 are not publicly available. As a PROTAC molecule with a molecular weight of 1092.4 g/mol, it may have poor oral bioavailability due to high molecular weight and the presence of multiple hydrogen bond donors/acceptors. The compound likely requires formulation with solubilizing agents (e.g., Kolliphor EL, PEG400) to improve exposure. In vivo half-life in rodents is expected to be short (1-4 hours) due to metabolism of the peptide-like linker. For quantitative PK analysis, an LC-MS/MS method would be developed to measure plasma concentrations. Key parameters such as Cmax, Tmax, AUC(0-∞), clearance, and volume of distribution would be determined following a single IV or oral dose. The compound is typically stored as a powder at -20degC and should be handled under inert conditions.
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| Toxicity/Toxicokinetics |
Toxicological data for PROTAC EGFR degrader 3 are not available in public sources. As a research compound, toxicity assessments are typically conducted in preliminary in vivo studies. Based on its mechanism of inducing protein degradation, potential toxicities could include on-target toxicities related to wild-type EGFR degradation in normal tissues (e.g., skin rash, diarrhea) as well as off-target degradation of other proteins. A typical acute toxicity study would involve a single ascending dose (e.g., 50, 150, 500 mg/kg) given orally to mice, followed by 14 days of observation for mortality, clinical signs, and body weight changes. Sub-chronic studies would involve daily dosing for 28 days, with serum chemistry, hematology, and histopathology of major organs (liver, kidney, heart, lung). No formal IND-enabling toxicology has been performed, as this is a research tool.
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| References | |
| Additional Infomation |
PROTAC EGFR degrader 3 is a research-grade chemical probe intended for laboratory use only and is not approved for clinical use in humans. It represents a novel approach to degrade mutant EGFR to overcome drug resistance in non-small cell lung cancer. The compound is available with high purity (typically ≥97%) and stored as a solid powder at -20degC. It is classified as a PROTAC and a protein degrader, and it is used in the field of targeted protein degradation (TPD). No clinical trials have been registered for this specific compound. It is part of a growing class of heterobifunctional molecules that harness the ubiquitin-proteasome system to eliminate disease-causing proteins. The compound's mechanism of lysosomal involvement in EGFR mutant degradation provides insights into alternative degradation pathways beyond the proteasome.
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| Molecular Formula |
C60H77N13O5S
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|---|---|
| Molecular Weight |
1092.40
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| Exact Mass |
1091.589
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| CAS # |
2768472-28-0
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| PubChem CID |
162513220
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| Appearance |
Off-white to light yellow solid powder
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| Density |
1.31±0.1 g/cm3(Predicted)
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| LogP |
0
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
22
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| Heavy Atom Count |
79
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| Complexity |
1970
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| Defined Atom Stereocenter Count |
5
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| SMILES |
CC1=C(SC=N1)C2=CC=C(C=C2)[C@H](C)NC(=O)[C@@H]3C[C@H](CN3C(=O)[C@H](C(C)(C)C)NC(=O)CCCCCCCN4CCN(CC4)C5=CC=C(C=C5)NC6=NC=C7C(=N6)N(C(=N7)NC8=CC=CC=C8)[C@H]9CCCN(C9)C(=O)C=C)O
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| InChi Key |
RYXQXEGTLNBKEC-DKQRTKIFSA-N
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| InChi Code |
InChI=1S/C60H77N13O5S/c1-7-52(76)71-30-16-19-47(37-71)73-55-49(66-59(73)65-44-17-12-11-13-18-44)36-61-58(68-55)64-45-25-27-46(28-26-45)70-33-31-69(32-34-70)29-15-10-8-9-14-20-51(75)67-54(60(4,5)6)57(78)72-38-48(74)35-50(72)56(77)63-40(2)42-21-23-43(24-22-42)53-41(3)62-39-79-53/h7,11-13,17-18,21-28,36,39-40,47-48,50,54,74H,1,8-10,14-16,19-20,29-35,37-38H2,2-6H3,(H,63,77)(H,65,66)(H,67,75)(H,61,64,68)/t40-,47-,48+,50-,54+/m0/s1
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| Chemical Name |
(2S,4R)-1-[(2S)-2-[8-[4-[4-[[8-anilino-9-[(3S)-1-prop-2-enoylpiperidin-3-yl]purin-2-yl]amino]phenyl]piperazin-1-yl]octanoylamino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~91.54 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.29 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.9154 mL | 4.5771 mL | 9.1542 mL | |
| 5 mM | 0.1831 mL | 0.9154 mL | 1.8308 mL | |
| 10 mM | 0.0915 mL | 0.4577 mL | 0.9154 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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