• Molecular & Cellular Toxicology
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• v.4 no.4
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• pp.323-330
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• 2008

Surrogate tissue analysis incorporating -omics technologies has emerged as a potential alternative method for evaluating toxic effect of the tissues which are not accessible for sampling. Among the recent applications, blood including whole blood, peripheral blood lymphocytes and peripheral blood mononuclear cells (PBMCs) was suggested as a suitable surrogate tissue in determining toxicant exposure and effect at the pre- or early clinical stage. In this application, we investigated transcriptomic profiles in astemizole treated Cynomolgus monkey's PBMCs. PBMCs were isolated from 4-6 years old male monkeys at 24 hr after administration45 Helvetica Light (10 mg/kg, 30 mg/kg). Gene expression profiles of astemizole treated monkey's PBMCs were determined using Affymetrix $GeneChip^{(R)}$ Human Genome U133 plus 2.0 arrays. The expression levels of 724 probe sets were significantly altered in PBMCs at 10 or 30 mg/kg after astemizole administration following determination of paired t-test using statistical criteria of ${\geq}$$1.5-fold changes at P<0.05. Gene expression patterns in PBMCs showed a considerable difference between astemizole 10 and 30 mg/kg administration groups in spite of an administration of the same chemical. However, close examination using Ingenuity Pathway Analysis (IPA) software revealed that several gene sets related to cardiotoxicity were deregulated at astemizole 10 and 30 mg/kg administration groups. The deregulation of cardiac hypertrophy related genes such as TXN, GNAQ, and MAP3K5 was observed at 10 mg/kg group. In astemizole 30 mg/kg group, genes involved in cardiotoxicity including cardiac necrosis/cell death, dilation, fibrosis, and hypertrophy were also identified. These results suggest that toxicogenomic approach using PBMCs as surrogate tissues will contribute to assess toxicant exposures and identify biomarkers at the pre-clinical stage.

• Toxicological Research
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• v.24 no.4
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• pp.289-295
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• 2008

Toxicology screening following treatment with astemizole, a histamine receptor antagonist, at oral doses of 0, 10, 30 and 60 mg/kg was carried out in male cynomolgus monkeys (Macaca fascicularis). No dose-related changes in mortality, clinical signs, body weight changes, food consumption, or urine analysis occurred in any animal compared to the vehicle control. However, the high-dose group showed a decrease in BUN and ALP compared to vehicle control group. In addition, the levels of TG, AST, ALP and CK increased. Although astemizole did not produce significant toxicological changes at any dose tested, we predict that it can cause toxicological changes of the liver and heart based on the changes in the serum parameters related to the heart and liver. The Action Potential Duration (APD) was prolonged in the heart of 60 mg/kg treatment group compared to the control group. The APD increase in 60 mg/kg treatment group along the other related changes in toxicological parameters imply that astemizole has major cardiotoxic effects in the cynomolgus monkey. This study is a valuable assessment for predicting the general toxicity and cardiotoxic effects of antihistamine drugs using nonhuman primates.

• YAKHAK HOEJI
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• v.43 no.5
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• pp.642-651
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• 1999

Nonsedating antihistamines do net cause sedation in therapeutic doses because these drugs hardly cross the blood-brain barrier. Since most of the peripheral side dffects of conventional antihistamines are related to their muscarinic receptor blocking action, the present study was performed to investigate whether nonsedating antihistamines interact with the muscarinic receptors and discriminate the muscarinic receptor subtypes in the rat cerebral microsomal fraction which containes both $M_1,{\;}M_2,{\;}M_3{\;}and{\;}M_4$ receptors. Five nonsedating antihistamines at high concentrations inhibited [$^3H$]QNB binding to the muscarinic receptor in a dose-dependent manner. The inhibition curves of these drugs except loratadine which showed positive cooperativity (nH=1.55) were steep (nH=1), indicating interaction with a single homogenous population of the binding sites. Astemizole, clemizole and mequitazine increased the $K_D$ value for [$^3H$]QNB without affecting the binding site concentrations, and this increase in the $K_D$ value resulted from the ability of these drugs to slow [$^3H$]QNB-receptor association. The Ki values of astemizole, clemizole and mequitazine for the inhibition for the inhibition of [$^3H$]QNB binding to muscarinic receptor were 0.58, 5.99 and $0.007{\;}{\mu}M$, respectively. However, loratadine and terfenadine inhibited noncompetitively [$^3H$]QNB binding with the normalized $IC_50$ value of about $2{\;}{\mu}M$. These results demonstrate that; 1) astemizole, clemizole and mequitazine interact directly with the muscarinic receptor at high concentrations; 2) muscarinic receptor blocking potency of these drugs varies widely among drugs; 3) these drugs do not discriminate between muscarinic receptor subtypes.

• Mass Spectrometry Letters
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• v.11 no.4
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• pp.113-117
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• 2020

Cytochrome P450 2J2 (CYP2J2) is a member of the cytochrome P450 superfamily, and is known to be arachidonic acid epoxygenase that mediates the formation of four bioactive regioisomers of epoxyeicosatrienoic acids (EETs). CYP2J2 is also involved in the metabolism of drugs such as albendazole, astemizole, danazol, ebastine, and terfenadine. CYP2J2 is highly expressed in the heart and cancer tissues. In this study, the inhibitory potential of ten natural products against CYP2J2 activity was evaluated using human liver microsomes and tandem mass spectrometry. Among them, bilobetin, which is a kind of biflavonoid, exhibits a strong inhibitory effect against the CYP2J2-mediated astemizole O-demethylation (IC50 = 0.73 μM) and terfenadine hydroxylation (IC50 = 0.89 μM). This result suggests that bilobetin can be used as strong CYP2J2 inhibitor in drug metabolism study.

• Journal of Life Science
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• v.23 no.9
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• pp.1126-1132
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• 2013

Cytochrome P450 2J2 (CYP2J2) is an enzyme mainly found in human extrahepatic tissues, with predominant expression in the cardiovascular system. CYP2J2 plays important roles in the metabolism of endogenous metabolites and therapeutic drugs, such as arachidonic acid, astemizole, ebastine, and terfenadine. CYP2J2 is also overexpressed in human cancer tissues and cancer cell lines and may represent a potential target for therapy of human cancers. In this study, 10 natural products obtained from plants and microorganisms were screened as potential CYP2J2 inhibitors. Among them, thelephoric acid showed strong inhibition of astemizole O-demethylation activity ($IC_{50}=3.23{\mu}M$) in a dose-dependent manner. Evaluation of the substrate dependency of the inhibitory activity of thelephoric acid showed that it strongly inhibited CYP2J2-mediated ebastine hydroxylation ($IC_{50}=5.32{\mu}M$) and terfenadine hydroxylation ($IC_{50}=3.27{\mu}M$) in a substrate nondependent manner. The present data suggest that this compound might be a potential candidate for further evaluation for anticancer activity.

• YAKHAK HOEJI
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• v.50 no.2
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• pp.129-135
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• 2006

Two Antihistamines, terfenadine and astemizole have been withdrawn from major markets for the reason that these durgs have been reported to induce QT interval prolongation associated with the onset of Torsades do Pointes (TdP), resulting in a life-threatening ventricular arrhythmia. In this study, we investigated effects of diphenhydramine on electrocardiograms and hemodynamic parameters in conscious telemetered dogs. We validated and defined the sensitivity of the test system by monitoring basal parameters and using positive control substance, terfenadine. Single administration effects were tested during 24 hours for each test drug at dose 1 mg/kg, 10 mg/kg, 30 mg/kg, 100 mg/kg. We monitored QT, QTc, heart rate, blood pressure and body temperature after administering test drugs. In conscious telemetered dogs, diphenhydramine significantly prolonged $QT\;(6.8\%\;of\;basal)\;and\;QTc\;(7.8\%\;of\;basal)$ at 100 mg/kg. Other parameters were not affected significantly. These findings suggest that antihistamines could induce important clinical relevance for patients taking excessive dosages of conventional antihistamines and those at risk of developing cardiac arrhythmias. Future studies that include other antihistamines and other classes will be necessary to predict the torsadogenic risk of drugs in humans.

• Journal of Life Science
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• v.21 no.11
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• pp.1558-1564
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• 2011

Cytochrome P450 2J2 (CYP2J2) plays important roles in the metabolism of endogenous metabolites such as arachidonic acid as well as therapeutic drugs. CYP2J2 is overexpressed in human cancer tissues and cancer cell lines, as well as in epoxyeicosatrienoic acids (EETs) and CYP2J2-mediated metabolites, and prevent apoptosis of cancer cells. This study aimed to screen marketed drugs for inhibitory potential on CYP2J2 isoforms using human liver microsomes. The initial screen isolated 4 compounds, from 120 marketed drugs, that inhibited the CYP2J2-mediated astemizole O-demethylation more than 50% in the following the order: haloperidol (75%) > terfenadine (56%) > aripiprazole (55%) > miconazole (52%). Miconazole strongly inhibited CYP2J2-mediated ebastine hydroxylation ($IC_{50}$=11.2 ${\mu}M$) and terfenadine hydroxylation ($IC_{50}$=2.2 ${\mu}M$), and terfenadine also inhibited CYP2J2-mediated ebastine hydroxylation ($IC_{50}$=13.6 ${\mu}M$) in a dose dependent manner. The present data suggest that these drugs are potential candidates for further evaluation for their anti-cancer activities.

• The Korean Journal of Gastroenterology
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• v.72 no.6
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• pp.325-328
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• 2018

위마비증(gastroparesis)이나 만성 구역 구토 증후군(chronic unexplained nausea and vomiting)으로 인한 구역과 구토에 대한 치료는 일반적으로 사용하는 위장관 촉진제에 만족스럽지 못한 경우가 많고, 여러 부작용으로 인하여 장기적으로 사용하기 어려워 보다 효과적인 치료 방법이 필요하다. 최근 미국에서 발표된 본 연구는 위마비증이나 연관 증후군 환자에서 구역과 구토 증상을 줄이기 위한 aprepitant(neurokinin-1 receptor antagonist) 치료의 효과를 분석한 것으로, 향후 일반적인 치료에 불응성 위마비증 환자에서 새로운 약제 사용을 시도해볼 수 있어 소개하고자 한다. 본 Aprepitant for the Relief of Nausea (APRON) 연구는 기질적 질환을 배제하기 위하여 최근 2년 이내 위내시경이 정상이며, 적어도 6개월 이상 조기 포만감(early satiety), 식후 만복감(postprandial fullness), 팽만감(bloating) 그리고 명치부 통증(epigastric pain)을 유발하는 증상과 함께 만성적인 구역이 있는 18세 이상의 성인 중 4시간의 위배출 검사를 시행받은 환자를 대상으로 하였다. 객관적인 지표로 0점에서 45점까지 보이는 9-증상 Gastroparesis Cardinal Symptom Index(GCSI)가 2주 이상 총 21점 이상이며, 0-100 mm의 visual analog scale (VAS)의 7일간의 구역 증상 평균 25 mm 이상인 환자를 대상으로 하였다. 일주일에 3일 이상 narcotics를 사용하였거나 와파린이나 pimozide, terfenadien, astemizole, cisapride를 복용하였던 환자, 2배 이상으로 간 효소 수치상승을 보이거나 Child-Pugh score 10점 이상, aprepitant에 알레르기를 보이는 환자는 제외되었다. 그렇지만 metoclopramide나 erythromycin을 안정적으로 사용 중인 환자는 제외되지 않았다. 위배출 검사는 2시간에서 60% 이상 남아 있거나 4시간에서 10% 이상 남아 있는 경우에 지연된 것으로 정의되었으며, 지연된 위배출 검사 결과 자체는 환자의 등록 기준에 포함되진 않았다. 등록 기준에 포함된 환자는 1:1로 무작위 배정되어 하루 한 번 125 mg의 aprepitant 복용군과 위약군으로 나누어져 연구가 진행되었으며, 약제 복용 4주간 2주 간격으로, 그리고 복용 후 2주 뒤까지 구역 증상의 호전 정도와 약제 안전성을 확인하였다. 이러한 효과를 판정하기 위하여 환자가 방문하는 동안 GCSI를 포함한 Patient Assessment of Upper GI Symptoms (PAGI-SYM), Gastrointestinal Symptom Rating Scale, daily VAS, daily diary version of the GCSI 그리고 정신 측정 도구와 삶의 질 도구인 Patient health Questionnaire 15와 Short Form 36 version이 측정되었다. 구역에 대한 aprepitant와 위약의 치료 효과의 일차적 판정은 이전 항암 요법에 대한 aprepitant 연구와 같이 28일 평균 VAS 25 mm 미만이거나 치료 전 7일간의 VAS와 비교하여 28일 치료 기간 동안 25 mm 이상 감소한 경우로 정의하였고, 이차 결과는 구역의 매일 시간, 치료 중 구역이 없는 날짜의 퍼센트, PAGI-SYM score의 개선 등으로 확인하였다. 2013년 4월부터 2015년 7월까지 총 126명의 환자가 등록되어 aprepitant군 63명, 위약군 63명으로 무작위 배정되었다. 전체의 57%인 72명에서 위배출 지연이 보였으며, 나머지 43%에서는 정상 또는 빠른 위배출 소견을 보여 만성적으로 설명할 수 없는 구역과 구토에 포함된 환자군으로 확인되었다. 또한 29%에서 당뇨를 가지고 있었으며, 8%에서 수면제를 사용하고 있었다. 최종적으로 aprepitant군은 59명, 위약군은 63명이 연구를 끝까지 종료하였다. 일차 결과에서 aprepitant 군 46%, 위약군 40%의 구역 호전을 보여 두 치료군 간에 통계적으로 의미 있는 차이는 보이지 않았다(상대 위험도 1.2, 95% CI: 0.8-1.7; p=0.43). 그러나 일차 분석의 두 가지 척도(28일 평균 VAS 25 mm 미만과 기저 VAS보다 평균 28일 VAS의 25 mm 이상 감소)를 모두 함께 고려한 민감도 분석에서는 aprepitant군이 37% (22/59)로 위약군의 17%(11/63)에 비하여 통계적으로 의미 있는 구역의 호전을 보였다(상대 위험도 2.1, 95% CI: 1.1-4.1; p=0.01). 또한 이차 분석을 살펴보면 aprepitant군에서 PAGI-SYM 중증도 지수로 확인하였을 때, 구역(1.8 vs. 1.0; p=0.005)과 구토(1.6 vs. 0.5; p=0.001)의 중증도 및 매일 구역 시간의 감소를 보였고, 28일 동안 구역이 없는 날짜의 퍼센트 증가 소견을 보였다. 다른 이차 결과 분석에서 aprepitant군이 PAGI-SYM 중증도 지수의 GCSI 종합 점수(1.3 vs. 0.7; p=0.001), 상당한 증상호전, 구역 구토의 세부 점수, 팽만감 세부 점수 그리고 위식도 역류 증상 점수에서 호전을 보였고, 매일 일기로 표현한 daily diary version of the GCSI에 상복부 통증 중증도, 전체 증상 그리고 Gastrointestinal Symptom Rating Scale의 종합 점수에서 호전을 보였다. 연구 중 발생한 부작용은 주로 경증과 중등도 정도의 부작용이 주로 발생하였지만, aprepitant군(35% vs. 17% 위약군, p=0.04)에서 더 많이 발생하였다. 결론적으로 위마비증 또는 위마비증 유사 증후군으로 인한 만성 구역 및 구토 환자의 무작위 시험에서 aprepitant는 VAS 점수를 통한 주요 결과를 분석하였을 때는 구역의 중증도를 호전시키지 못하였지만 다른 이차적 결과에 대해서는 위약군에 대하여 호전 소견을 보였다. 따라서 aprepitant에 효과적인 반응을 보이는 위마비증 환자를 감별하는 추가 임상시험이 필요할 것으로 판단된다.