• BMB Reports
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• 제50권3호
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• pp.150-155
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• 2017

Non-small-cell lung cancer (NSCLC) is the third most common cancer that spreads to the bone, resulting in osteolytic lesions caused by hyperactivation of osteoclasts. Activating mutations in epidermal growth factor receptor-tyrosine kinase (EGF-TK) are frequently associated with NSCLC, and afatinib is a first-line therapeutic drug, irreversibly targeting EGF-TK. However, the effects of afatinib on osteoclast differentiation and activation as well as the underlying mechanism remain unclear. In this study, afatinib significantly suppressed receptor activator of nuclear factor ${\kappa}B$ (RANK) ligand (RANKL)-induced osteoclast formation in bone marrow macrophages (BMMs). Consistently, afatinib inhibited the expression of osteoclast marker genes, whereas, it upregulated the expression of negative modulator genes. The bone resorbing activity of osteoclasts was also abrogated by afatinib. In addition, afatinib significantly inhibited RANKL-mediated Akt/protein kinase B and c-Jun N-terminal kinase phosphorylation. These results suggest that afatinib substantially suppresses osteoclastogenesis by downregulating RANK signaling pathways, and thus may reduce osteolysis after bone metastasis.

• Tuberculosis and Respiratory Diseases
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• 제85권2호
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• pp.147-154
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• 2022

Background: The expression of calcium signaling pathway molecules is altered in various carcinomas, which are related to the proliferation and altered characteristics of cancer cells. However, changes in calcium signaling in anti-cancer drug-resistant cells (bearing a T790M mutation in epidermal growth factor receptor [EGFR]) remain unclear. Methods: Afatinib-mediated changes in the level of store-operated Ca²⁺ entry (SOCE)-related proteins and intracellular Ca²⁺ level in non-small cell lung cancer cells with T790M mutation in the EGFR gene were analyzed using western blot and ratiometric assays, respectively. Afatinib-mediated autophagic flux was evaluated by measuring the cleavage of LC3B-II. Flow cytometry and cell proliferation assays were conducted to assess cell apoptosis and proliferation. Results: The levels of SOCE-mediating proteins (ORAI calcium release-activated calcium modulator 1 [ORAI1], stromal interaction molecule 1 [STIM1], and sarco/endoplasmic reticulum Ca²⁺ ATPase [SERCA2]) decreased after afatinib treatment in non-small cell lung cancer cells, whereas the levels of SOCE-related proteins did not change in gefitinib-resistant non-small cell lung cancer cells (PC-9/GR; bearing a T790M mutation in EGFR). Notably, the expression level of SOCE-related proteins in PC-9/GR cells was reduced also responding to afatinib in the absence of extracellular Ca²⁺. Moreover, extracellular Ca²⁺ influx through the SOCE was significantly reduced in PC-9 cells pre-treated with afatinib than in the control group. Additionally, afatinib was found to decrease the level of SOCE-related proteins through autophagic degradation, and the proliferation of PC-9GR cells was significantly inhibited by a lack of extracellular Ca²⁺. Conclusion: Extracellular Ca²⁺ plays important role in afatinib-mediated autophagic degradation of SOCE-related proteins in cells with T790M mutation in the EGFR gene and extracellular Ca²⁺ is essential for determining anti-cancer drug efficacy.

• 대한한방내과학회지
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• 제38권1호
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• pp.72-80
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• 2017

Objective: The purpose of this study is to report the case of a patient with recurring non-small cell lung carcinoma (NSCLC) taking Samchilchoongcho-Jung with Afatinib. Methods: An NSCLC patient diagnosed with multiple bone and pulmonary metastasis was taking Afatinib (20 mg/day) and suffering from stomatitis caused by the Afatinib. The patient was treated with Samchilchoongcho-Jung (1,500 mg/ day) for 3 months. The tumor size was measured with computed tomography, and laboratory findings, including tumor markers (CEA, Cyfra 21-1), were also followed up. Stomatitis was measured by a numeric rating scale, and adverse events were evaluated using the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI-CTCAE), version 4.0. Results: After combined treatment, stable disease was shown on computed tomography. The tumor marker levels of CEA and Cyfra 21-1 were decreased, and the stomatitis significantly improved. NCI-CTCAE 4.0 showed no adverse events. Conclusion: This case study suggests that Samchilchoongcho-Jung may have a synergic effect, in conjunction with Afatinib, on the treatment of patients with recurring NSCLC.

• 대한한방내과학회지
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• 제41권6호
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• pp.1255-1264
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• 2020

Objective: The purpose of this study was to report the effect of Korean medicine on a patient with non-small cell lung cancer with pleural metastasis who had been treated with afatinib. Method: A 61-year old female patient with non-small cell lung cancer with pleural metastasis was treated with acupuncture and herbal medicines, including Yijung-tang, Haengso-tang, Samchulkunbi-tang, Paeamju-bang (Feiai zhu fang), to control various symptoms caused by afatinib. The degree of pain was assessed by a numeric rating scale (NRS) and the quality of life was determined with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, Core 30 (EORTC QLQ-C30) and the EORTC 13-item lung cancer-specific module (EORTC LC-13). Results: After receiving acupuncture and herbal medicines, the patient showed improvement in her back and chest pain, according to the NRS score. Similarly, the Korean medicinal treatments significantly relieved her nausea, vomiting, diarrhea, hemoptysis, and alopecia. However, the EORTC QLQ-C30 assessment suggested that Korean medicinal treatments did not significantly improve the global health status of this patient. Conclusion: Korean medicine could be useful in relieving some of the symptoms occurring after conventional afatinib treatments.

• Parasites, Hosts and Diseases
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• 제54권1호
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• pp.31-38
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• 2016

Specific gene expressions of host cells by spontaneous STAT6 phosphorylation are major strategy for the survival of intracellular Toxoplasma gondii against parasiticidal events through STAT1 phosphorylation by infection provoked $IFN-{\gamma}$. We determined the effects of small molecules of tyrosine kinase inhibitors (TKIs) on the growth of T. gondii and on the relationship with STAT1 and STAT6 phosphorylation in ARPE-19 cells. We counted the number of T. gondii RH tachyzoites per parasitophorous vacuolar membrane (PVM) after treatment with TKIs at 12-hr intervals for 72 hr. The change of STAT6 phosphorylation was assessed via western blot and immunofluorescence assay. Among the tested TKIs, Afatinib (pan ErbB/EGFR inhibitor, $5{\mu}M$) inhibited 98.0% of the growth of T. gondii, which was comparable to pyrimethamine ($5{\mu}M$) at 96.9% and followed by Erlotinib (ErbB1/EGFR inhibitor, $20{\mu}M$) at 33.8% and Sunitinib (PDGFR or c-Kit inhibitor, $10{\mu}M$) at 21.3%. In the early stage of the infection (2, 4, and 8 hr after T. gondii challenge), Afatinib inhibited the phosphorylation of STAT6 in western blot and immunofluorescence assay. Both JAK1 and JAK3, the upper hierarchical kinases of cytokine signaling, were strongly phosphorylated at 2 hr and then disappeared entirely after 4 hr. Some TKIs, especially the EGFR inhibitors, might play an important role in the inhibition of intracellular replication of T. gondii through the inhibition of the direct phosphorylation of STAT6 by T. gondii.

• 대한한방내과학회지
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• 제39권5호
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• pp.973-983
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• 2018

Objectives: We report a case of traditional Korean medicine (TKM) treatment for skin side effects after taking afatinib (Giotrif$^{(R)}$). Methods: A 62-year-old female who was diagnosed with non-small cell lung cancer stage 4 (T4N2M1b) and was on treatment with afatinib (29.56 mg/day for 4 months) complained of skin toxicity as a side effect. For 16 admission days, the patient was treated with acupuncture, moxibustion, and herbal medicine (oral decoction and external ointment). Results: Improvement of skin toxicity was measured by a numeric rating scale. In addition, Quality of life (QOL) was measured using EORTC Quality of Life Questionnaire, Core 30 (EORTC QLQ-C30) and EORTC Quality of Life Questionnaire, 13-item lung cancer-specific module (EORTC QLQ-LC13) Developed by the European Organization for Research and Treatment of Cancer (EORTC). Tumor size and carcinoembryonic antigen (CEA) were also examined during follow up. Conclusions: After a combined TKM treatment, skin toxicity symptoms and quality of life scales were significantly improved with no side effects. The tumor size was not changed on computed tomography during follow-up period. CEA levels were decreased.

• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8191-8196
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• 2016

Inhibition of EGFR-EGF interactions forms an important therapeutic rationale in treatment of non-small cell lung carcinoma. Established inhibitors have been successful in reducing proliferative processes observed in NSCLC, however patients suffer serious side effects. Considering the narrow therapeutic window of present EGFR inhibitors, the present study centred on identifying high efficacy EGFR inhibitors through structure based virtual screening strategies. Established inhibitors - Afatinib, Dacomitinib, Erlotinib, Lapatinib, Rociletinib formed parent compounds to retrieve similar compounds by linear fingerprint based tanimoto search with a threshold of 90%. The compounds (parents and respective similars) were docked at the EGF binding cleft of EGFR. Patch dock supervised protein-protein interactions were established between EGF and ligand (query and similar) bound and free states of EGFR. Compounds ADS103317, AKOS024836912, AGN-PC-0MXVWT, GNF-Pf-3539, SCHEMBL15205939 were retrieved respectively similar to Afatinib, Dacomitinib, Erlotinib, Lapatinib, Rociletinib. Compound-AGN-PC-0MXVWT akin to Erlotinib showed highest affinity against EGFR amongst all the compounds (parent and similar) assessed in the study. Further, AGN-PC-0MXVWT brought about significant blocking of EGFR-EGF interactions in addition showed appreciable ADMET properties and pharmacophoric features. In the study, we report AGN-PC-0MXVWT to be an efficient and high efficacy inhibitor of EGFR-EGF interactions identified through computational approaches.

• Parasites, Hosts and Diseases
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• 제58권3호
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• pp.249-255
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• 2020

Toxoplasma gondii, a ubiquitous, intracellular parasite of the phylum Apicomplexa, infects an estimated one-third of the human population as well as a broad range of warm-blooded animals. We have observed that some tyrosine kinase inhibitors suppressed the growth of T. gondii within host ARPE-10 cells. Among them, afatinib, human epithermal growth factor receptor 2 and 4 (HER2/4) inhibitor, may be used as a therapeutic agent for inhibiting parasite growth with minimal adverse effects on host. In this report, we conducted a proteomic analysis to observe changes in host proteins that were altered via infection with T. gondii and the treatment of HER2/4 inhibitors. Secreting proteins were subjected to a procedure of micor basic reverse phase liquid chromatography, nano-liquid chromatography-mass spectrometry, and ingenuity pathway analysis serially. As a result, the expression level of heterogeneous nuclear ribonucleoprotein K, semaphorin 7A, a GPI membrane anchor, serine/threonine-protein phosphatase 2A, and calpain small subunit 1 proteins were significantly changed, and which were confirmed further by western blot analysis. Changes in various proteins, including these 4 proteins, can be used as a basis for explaining the effects of T. gondii infections and HER2/4 inhibitors.

• Biomolecules & Therapeutics
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• 제30권1호
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• pp.19-27
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• 2022

Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase widely expressed in many cancers such as non-small cell lung cancer (NSCLC), pancreatic cancer, breast cancer, and head and neck cancer. Mutations such as L858R in exon 21, exon 19 truncation (Del19), exon 20 insertions, and others are responsible for aberrant activation of EGFR in NSCLC. First-generation EGFR tyrosine kinase inhibitors (TKIs) such as gefitinib and erlotinib have clinical benefits for EGFR-sensitive (L858R and Del19) NSCLC patients. However, after 10-12 months of treatment with these inhibitors, a secondary T790M mutation at the gatekeeper position in the kinase domain of EGFR was identified, which limited the clinical benefits. Second-generation EGFR irreversible inhibitors (afatinib and dacomitinib) were developed to overcome this T790M mutation. However, their lack of selectivity toward wild-type EGFR compromised their clinical benefits due to serious adverse events. Recently developed third-generation irreversible EGFR TKIs (osimertinib and lazertinib) are selective toward driving mutations and the T790M mutation, while sparing wild-type EGFR activity. The latest studies have concluded that their efficacy was also compromised by additional acquired mutations, including C797S, the key residue cysteine that forms covalent bonds with irreversible inhibitors. Because second- and third-generation EGFR TKIs are irreversible inhibitors, they are not effective against C797S containing EGFR triple mutations (Del19/T790M/C797S and L858R/T790M/C797S). Therefore, there is an urgent unmet medical need to develop next-generation EGFR TKIs that selectively inhibit EGFR triple mutations via a non-irreversible mechanism.

• 대한암한의학회지
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• 제23권1호
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• pp.23-32
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• 2018

ALK 변이는 비소세포성 폐암에서 차지하는 비율은 높지 않지만 비소세포성 폐암의 발생률이 높기 때문에 환자 수는 적지 않다. ALK 저해제인 Crizotinib은 환자들의 무진행 생존을 평균 대략 4개월 정도 연장시키고 증상을 완화시키며 항암 치료를 받는 것에 비교하여 삶의 질을 향상시키는 치료성과를 거두었다. 하지만 약물 내성 발현은 주요 한계이며 여전히 ALK 변이 비소세포성 폐암 환자의 예후가 불량하므로 내성을 극복하고 지속적인 치료 반응을 유도하여 치료율을 상승시키기 위한 연구가 필요하다. 현재 폐암치료 한약제제인 삼칠충초정과 2세대 EGFR TKI 제제인 Afatinib의 동시 사용 시 상승효과가 나타난다는 연구결과는 보고되어 있지만 ALK 저해제인 Crizotinib과의 병용 치료와 관련한 연구는 시행된 바가 없었다. 본 증례는 삼칠충초정과 Crizotinib의 병용 투여를 통해 종양 크기의 불변 및 삶의 질 개선과 Crizotinib의 내성 억제 가능성 등을 보인 점에서 의의가 있으나 대조군이 없는 1례에 불과하여 Crizotinib의 단독 효과일 가능성을 배제할 수 없는 한계를 지닌다. 따라서 향후 삼칠충초정과 Crizotinib의 병용투여에 대한 추가적인 연구 및 임상시험을 통해 더욱 객관적인 효과 판정이 필요할 것으로 사료된다.