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Discovery and characterization of a novel irreversible EGFR mutants selective and potent kinase inhibitor CHMFL-EGFR-26 with a distinct binding mode

View Article: PubMed Central - PubMed

ABSTRACT

EGFR T790M mutation accounts for about 40-55% drug resistance for the first generation EGFR kinase inhibitors in the NSCLC. Starting from ibrutinib, a highly potent irreversible BTK kinase inhibitor, which was also found to be moderately active to EGFR T790M mutant, we discovered a highly potent irreversible EGFR inhibitor CHMFL-EGFR-26, which is selectively potent against EGFR mutants including L858R, del19, and L858R/T790M. It displayed proper selectivity window between the EGFR mutants and the wide-type. CHMFL-EGFR-26 exhibited good selectivity profile among 468 kinases/mutants tested (S score (1)=0.02). In addition, X-ray crystallography revealed a distinct “DFG-in” and “cHelix-out” inactive binding mode between CHMFL-EGFR-26 and EGFR T790M protein. The compound showed highly potent anti-proliferative efficacy against EGFR mutant but not wide-type NSCLC cell lines through effective inhibition of the EGFR mediated signaling pathway, induction of apoptosis and arresting of cell cycle progression. CHMFL-EGFR-26 bore acceptable pharmacokinetic properties and demonstrated dose-dependent tumor growth suppression in the H1975 (EGFR L858R/T790M) and PC-9 (EGFR del19) inoculated xenograft mouse models. Currently CHMFL-EGFR-26 is undergoing extensive pre-clinical evaluation for the clinical trial purpose.

No MeSH data available.


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Anti-tumor efficacy of CHMFL-EGFR-26 in H1975 cell inoculated xenograft mouse modeFemale nu/nu mice bearing established H1975 tumor xenografts were treated with CHMFL-EGFR-26 at 25.0, 50.0 and 100 mg/kg/d, or vehicle. Daily oral administration was initiated when H1975 tumors had reached a size of 200 to 400 mm3. Each group contained 5 or 6 animals. Data, mean ± SEM. A. Body weight and B. Tumor size measurements from H1975 xenograft mice after CHMFL-EGFR-26 administration. Initial body weight and tumor size were set as 100%. C. Representative photographs of tumors in each group after 25.0, 50.0 or 100 mg/kg/d CHMFL-EGFR-26 or vehicle treatment. D. Comparison of the final tumor weight in each group after 21-day treatment period. Numbers in columns indicate the mean tumor weight in each group. **p<0.01. E. Representative micrographs of hematoxylin and eosin (HE), Ki-67, and TUNEL staining of tumor tissues with CHMFL-EGFR-26 treatment compared to the vehicle-treated group. Note the specific nuclear staining of cells with morphology consistent with proliferation and apoptosis (E, red arrow).
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Figure 4: Anti-tumor efficacy of CHMFL-EGFR-26 in H1975 cell inoculated xenograft mouse modeFemale nu/nu mice bearing established H1975 tumor xenografts were treated with CHMFL-EGFR-26 at 25.0, 50.0 and 100 mg/kg/d, or vehicle. Daily oral administration was initiated when H1975 tumors had reached a size of 200 to 400 mm3. Each group contained 5 or 6 animals. Data, mean ± SEM. A. Body weight and B. Tumor size measurements from H1975 xenograft mice after CHMFL-EGFR-26 administration. Initial body weight and tumor size were set as 100%. C. Representative photographs of tumors in each group after 25.0, 50.0 or 100 mg/kg/d CHMFL-EGFR-26 or vehicle treatment. D. Comparison of the final tumor weight in each group after 21-day treatment period. Numbers in columns indicate the mean tumor weight in each group. **p<0.01. E. Representative micrographs of hematoxylin and eosin (HE), Ki-67, and TUNEL staining of tumor tissues with CHMFL-EGFR-26 treatment compared to the vehicle-treated group. Note the specific nuclear staining of cells with morphology consistent with proliferation and apoptosis (E, red arrow).

Mentions: In H1975 cells inoculated xenograft mouse model, oral administration of CHMFL-EGFR-26 with different dosages (25, 50 and 100mg/kg/day) did not show any apparent toxicity. (Figure 4A) It also exhibited dose-dependent tumor growth suppression and 100 mg/kg/day dosage could almost completely blocked the tumor progression and exhibited a TGI (tumor growth inhibition rate) of 60.1% (Figure 4B, 4C, 4D). Immunohistochemical (IHC) staining showed that CHMFL-EGFR-26 inhibited the cell proliferation by Ki67 stain and induced apoptosis by TUNEL stain in the tumor tissues. (Figure 4E) Similar results were observed in PC-9 cells inoculated xenograft mouse model. (Figure 5)


Discovery and characterization of a novel irreversible EGFR mutants selective and potent kinase inhibitor CHMFL-EGFR-26 with a distinct binding mode
Anti-tumor efficacy of CHMFL-EGFR-26 in H1975 cell inoculated xenograft mouse modeFemale nu/nu mice bearing established H1975 tumor xenografts were treated with CHMFL-EGFR-26 at 25.0, 50.0 and 100 mg/kg/d, or vehicle. Daily oral administration was initiated when H1975 tumors had reached a size of 200 to 400 mm3. Each group contained 5 or 6 animals. Data, mean ± SEM. A. Body weight and B. Tumor size measurements from H1975 xenograft mice after CHMFL-EGFR-26 administration. Initial body weight and tumor size were set as 100%. C. Representative photographs of tumors in each group after 25.0, 50.0 or 100 mg/kg/d CHMFL-EGFR-26 or vehicle treatment. D. Comparison of the final tumor weight in each group after 21-day treatment period. Numbers in columns indicate the mean tumor weight in each group. **p<0.01. E. Representative micrographs of hematoxylin and eosin (HE), Ki-67, and TUNEL staining of tumor tissues with CHMFL-EGFR-26 treatment compared to the vehicle-treated group. Note the specific nuclear staining of cells with morphology consistent with proliferation and apoptosis (E, red arrow).
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Figure 4: Anti-tumor efficacy of CHMFL-EGFR-26 in H1975 cell inoculated xenograft mouse modeFemale nu/nu mice bearing established H1975 tumor xenografts were treated with CHMFL-EGFR-26 at 25.0, 50.0 and 100 mg/kg/d, or vehicle. Daily oral administration was initiated when H1975 tumors had reached a size of 200 to 400 mm3. Each group contained 5 or 6 animals. Data, mean ± SEM. A. Body weight and B. Tumor size measurements from H1975 xenograft mice after CHMFL-EGFR-26 administration. Initial body weight and tumor size were set as 100%. C. Representative photographs of tumors in each group after 25.0, 50.0 or 100 mg/kg/d CHMFL-EGFR-26 or vehicle treatment. D. Comparison of the final tumor weight in each group after 21-day treatment period. Numbers in columns indicate the mean tumor weight in each group. **p<0.01. E. Representative micrographs of hematoxylin and eosin (HE), Ki-67, and TUNEL staining of tumor tissues with CHMFL-EGFR-26 treatment compared to the vehicle-treated group. Note the specific nuclear staining of cells with morphology consistent with proliferation and apoptosis (E, red arrow).
Mentions: In H1975 cells inoculated xenograft mouse model, oral administration of CHMFL-EGFR-26 with different dosages (25, 50 and 100mg/kg/day) did not show any apparent toxicity. (Figure 4A) It also exhibited dose-dependent tumor growth suppression and 100 mg/kg/day dosage could almost completely blocked the tumor progression and exhibited a TGI (tumor growth inhibition rate) of 60.1% (Figure 4B, 4C, 4D). Immunohistochemical (IHC) staining showed that CHMFL-EGFR-26 inhibited the cell proliferation by Ki67 stain and induced apoptosis by TUNEL stain in the tumor tissues. (Figure 4E) Similar results were observed in PC-9 cells inoculated xenograft mouse model. (Figure 5)

View Article: PubMed Central - PubMed

ABSTRACT

EGFR T790M mutation accounts for about 40-55% drug resistance for the first generation EGFR kinase inhibitors in the NSCLC. Starting from ibrutinib, a highly potent irreversible BTK kinase inhibitor, which was also found to be moderately active to EGFR T790M mutant, we discovered a highly potent irreversible EGFR inhibitor CHMFL-EGFR-26, which is selectively potent against EGFR mutants including L858R, del19, and L858R/T790M. It displayed proper selectivity window between the EGFR mutants and the wide-type. CHMFL-EGFR-26 exhibited good selectivity profile among 468 kinases/mutants tested (S score (1)=0.02). In addition, X-ray crystallography revealed a distinct &ldquo;DFG-in&rdquo; and &ldquo;cHelix-out&rdquo; inactive binding mode between CHMFL-EGFR-26 and EGFR T790M protein. The compound showed highly potent anti-proliferative efficacy against EGFR mutant but not wide-type NSCLC cell lines through effective inhibition of the EGFR mediated signaling pathway, induction of apoptosis and arresting of cell cycle progression. CHMFL-EGFR-26 bore acceptable pharmacokinetic properties and demonstrated dose-dependent tumor growth suppression in the H1975 (EGFR L858R/T790M) and PC-9 (EGFR del19) inoculated xenograft mouse models. Currently CHMFL-EGFR-26 is undergoing extensive pre-clinical evaluation for the clinical trial purpose.

No MeSH data available.


Related in: MedlinePlus