Determination of intracellular anlotinib, osimertinib, afatinib and gefitinib accumulations in human brain microvascular endothelial cells by liquid chromatography/tandem mass spectrometry

Abstract

Rationale Brain metastases are a common complication in patients with non‐small‐cell lung cancer (NSCLC). Anlotinib hydrochloride is a novel multi‐target tyrosine kinase inhibitor (TKI) exhibiting a superior overall response rate for brain metastases from NSCLC. The penetrability of anlotinib and three generations of epidermal growth factor receptor (EGFR) TKIs (osimertinib, afatinib and gefitinib) into brain microvascular endothelial cells (HBMECs) was compared. Methods A sensitive quantification method for the four TKIs was developed using liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). Anlotinib and the three EGFR TKIs were separated on an ACQUITY BEH C18 column after a direct protein precipitation, and then analyzed using electrospray ionization in positive ion mode. The linearity, accuracy, precision, limit of quantification, specificity and stability were assessed. Results The four analytes could be efficiently quantified in a single run of 3.8 min. The validation parameters of all analytes satisfy the acceptance criteria of bioanalytical method guidelines. The calibration range was 0.2–200 ng mL−1 for anlotinib and gefitinib, 1–500 ng mL−1 for osimertinib and 1–200 ng mL−1 for afatinib. The penetration of anlotinib across HBMECs was comparable with that of afatinib and gefitinib but less than that of osimertinib. Conclusions A sensitive LC/MS/MS method to simultaneously measure anlotinib, osimertinib, afatinib and gefitinib in cell extracts was successfully validated and applied to determine their uptake inside HBMECs, which could pave the way for future research on the role of anlotinib in NSCLC brain metastases.

Radiotherapy plus temozolomide with or without anlotinib in H3K27M‐mutant diffuse midline glioma: A retrospective cohort study

“…Currently, anlotinib has been approved by China National Medical Products Administration for the treatment of nonsmall‐cell lung cancer (NSCLC), small cell lung cancer, soft tissue sarcoma, and medullary thyroid cancer. In addition, anlotinib could cross the BBB confirmed by in vivo experiments and advanced NSCLC with brain metastasis 21 . On the other hand, radiotherapy could increase the permeability of blood–brain barrier and increase anlotinib concentration in brain tissue.…”

“…Previous studies reported favorable responses in NSCLC patients with brain metastases, demonstrating its ability to control brain tumors and survival. 22,47 Furthermore, Ma et al 21 confirmed optimal penetration of anlotinib into brain microvascular endothelial cells, potentially overcoming the limited drug concentration in brain tumors due to the blood-brain barrier (BBB). These findings suggest that anlotinib is a promising candidate for treating intracranial malignant tumors.…”

“…In addition, anlotinib could cross the BBB confirmed by in vivo experiments and advanced NSCLC with brain metastasis. 21 On the other hand, radiotherapy could increase the permeability of blood–brain barrier and increase anlotinib concentration in brain tissue. Thus, combining radiotherapy with anlotinib would benefit patients with brain metastases and gliomas.…”

Anlotinib plus Sintilimab achieved in an antitumor effect of complete remission in a patient with advanced hepatocellular carcinoma: a case report

“…There were studies that revealed that anlotinib potentially inhibits more targets and is superior to other antiangiogenesis drugs such as sunitinib, sorafenib and nintedanib both in vitro and in vivo [7]. Another confirmed advantage of anlotinib includes the better penetration of brain microvascular endothelial cells than other TKIs, including osimertinib, afatinib and gefitinib [16]. Compared with other common VEGFR-TKIs, anlotinib was reported to have a lower incidence of grade 3/4 adverse effects, which include thrombocytopenia, neutropenia including, hypertension, hand-foot syndrome, diarrhea, stomatitis and anorexia [17][18][19].…”

EGFR signaling pathway as therapeutic target in human cancers

“…In the future, with the promise of highly specific, possibly mutation-specific drugs, it will be possible to develop personalized drug treatments targeted to the molecular constitution of individual tumors [225]. In addition, state-of-the-art technologies, such as liquid chromatography coupled to tandem mass spectrometry, currently capable of simultaneously measuring multiple drugs in cell extracts [226], could also be adopted for in vivo follow-ups of specific drugs' uptakes in both primary and metastatic tumors, to better dose therapeutic treatments in a personalized manner.…”