Improved oral absorption of cilostazol via sulfonate salt formation with mesylate and besylate

Seo, Jae Hong,Park, Jung Bae,Choi, Woong-Kee,Park, Sunhwa,Sung, Yun Jin,Oh, Euichaul,Bae, Soo Kyung Dove Medical Press 2015 Drug design, development and therapy Vol.9 No.-

<P><B>Objective</B></P><P>Cilostazol is a Biopharmaceutical Classification System class II drug with low solubility and high permeability, so its oral absorption is variable and incomplete. The aim of this study was to prepare two sulfonate salts of cilostazol to increase the dissolution and hence the oral bioavailability of cilostazol.</P><P><B>Methods</B></P><P>Cilostazol mesylate and cilostazol besylate were synthesized from cilostazol by acid addition reaction with methane sulfonic acid and benzene sulfonic acid, respectively. The salt preparations were characterized by nuclear magnetic resonance spectroscopy. The water contents, hygroscopicity, stress stability, and photostability of the two cilostazol salts were also determined. The dissolution profiles in various pH conditions and pharmacokinetic studies in rats were compared with those of cilostazol-free base.</P><P><B>Results</B></P><P>The two cilostazol salts exhibited good physicochemical properties, such as nonhygroscopicity, stress stability, and photostability, which make it suitable for the preparation of pharmaceutical formulations. Both cilostazol mesylate and cilostazol besylate showed significantly improved dissolution rate and extent of drug release in the pH range 1.2–6.8 compared to the cilostazol-free base. In addition, after oral administration to rats, cilostazol mesylate and cilostazol besylate showed increases in <I>C</I><SUB>max</SUB> and AUC<I><SUB>t</SUB></I> of approximately 3.65- and 2.87-fold and 3.88- and 2.94-fold, respectively, compared to cilostazol-free base.</P><P><B>Conclusion</B></P><P>This study showed that two novel salts of cilostazol, such as cilostazol mesylate and cilostazol besylate, could be used to enhance its oral absorption. The findings warrant further preclinical and clinical studies on cilostazol mesylate and cilostazol besylate at doses lower than the usually recommended dosage, so that it can be established as an alternative to the marketed cilostazol tablet.</P>

렛트를 이용한 심적환과 cilostazol에 관한 상호작용 연구

김익균(Ekyune Kim) 한국생명과학회 2016 생명과학회지 Vol.26 No.1

본 연구의 최종 목표는 한방제제인 심적환과 심혈관계 양방 치료제인 cilostazol과 병용 투여 효과에 대한 포괄적이고 통합의학 측면에서 정확한 정보를 얻는 것이다. Cilostazol은 말초 동맥질환 치료제로 개발 된 항 혈소판 및 혈관확장제이다. 사이클릭 AMP protein kinase A를 활성 시켜 세포내 사이클릭 AMP (cAMP) 증가를 통하여 내피세포의 NO생산을 활성화 시킨다. 심적환을 단 회 또는 반복 투여 후 cilostazol의 약물 동태학적 효과를 평가하기 위하여 순수한 증류수 단회 용량과 증류수에 심적환 콜로이드 현탁액을 각각 대조군과 시험군에 투여 하여 30분 후, 두 그룹에 cilostazol를 투여하였다. 혈청은 cilostazol 약물 투여 30분 전에 수집 하였으며, cilostazol 약물 처리 후 0.25, 0.5, 0.45 및 1, 2, 4, 6, 8, 24시간 후에 각각 수집 하였다. 그 다음 실험군과 및 테스트 그룹 사이에 실로 스타 졸에서 관찰 된 약동학 적 변화를 평가 하였다. 통계적으로 유의 한 차이는 심적관 단독 투여와 반복투여군 그룹의 약물 동태 학에서 관찰되지 않았다. 이러한 연구 결과는 만성 질환 환자에서 한약제인 심적환의 투여는 cilostazol의 약동학에 영향을 미치지 않았음을 보여 주었다. 본 연구에서 얻어진 결과는 만성 혈관질환 환자에서 심적환과 cilostazol의 병용 투여를 제안하며 두 약물간의 잠재적 인 약물 상호 작용에 대한 cilostazol의 생체 이용률에 영향을 미치지 않을 것이라 판단된다. The object of this study was to obtain accurate information about the co-administration effects of cardiotonic pills on the pharmacokinetics of cilostazol were observed as a process of the comprehensive and integrative medicine. Cilostazol is a synthetic anti-platelet and vasodilator agent developed for the treatment of intermittent claudication resulting from peripheral arterial disease. By increasing intracellular cyclic adenosine monophosphate (cAMP), cilostazol induces the activation of protein kinase A, which activates endothelial nitric oxide synthase. In order to evaluate the effect of a single or repeated cardiotonic pill dose on the pharmacokinetics of cilostazol, a single dose of pure_distilled water or a colloidal suspension of distilled water and cardiotonic pills were administered to the control and test groups, respectively. After 30 min, both groups were administered cilostazol. Plasma was collected 30min before administration, and 0.25, 0.5, 0.45, 1, 2, 4, 6, 8, and 24h after the end of cilostazol treatment. We then evaluated the pharmacokinetic changes observed with cilostazol between the control and test groups. No statistically significant differences were observed. These findings demonstrated that a single dose of cardiotonic pills did not affect the pharmacokinetics of cilostazol. The results obtained in this study suggest that co-administration of cardiotonic pills and cilostazol may not affect the bioavailability of cilostazol as a potential drug interaction.

Monitoring the Antiplatelet Effect of Cilostazol with Light Transmission Aggregometer: Two Cases of Possible Cilostazol Resistance

심효은,장성수,박찬정,지현숙,이승환,박성욱 대한진단검사의학회 2016 Laboratory Medicine Online Vol.6 No.4

Background: Coronary artery disease is an important cause of death in adults and stent insertion is one of the treatment modalities. The most severe adverse effect of a stent insertion is the formation of a thrombus; therefore, antiplatelet agents are used. The addition of cilostazol to low-dose aspirin and clopidogrel results in a better antiplatelet effect. However, laboratory tests to monitor the effect of cilostazol are insufficient. Methods: We tested the inhibitory effect of cilostazol using maximal platelet aggregation in 20 healthy volunteers. Conditions for incubation and concentrations of cilostazol and prostaglandin E1 (PGE1) were established and aggregation was induced by 5´-adenosine diphosphate (ADP) and measured with light transmission aggregometry (LTA). Blood samples were incubated with 1 μM and 2 μM cilostazol for 10 minutes at room temperature, and 80 nM PGE1 was added and incubated for an additional 10 minutes. Aggregation was induced by ADP and reactivity was evaluated. Results: The average maximum aggregation (MA) was 58.1% at 1 μM cilostazol and 22.0% when PGE1 was added. The average MA was 42.8% when cilostazol concentration was increased to 2 μM and 21.2% when PGE1 was added. Average inhibition of aggregation at 1 μM cilostazol was not statistically significant (P=0.085), but was significant (P=0.004) at 2 μM cilostazol. Aggregation was not inhibited even with 2 μM cilostazol and PGE1 in 2 volunteers, which suggests possible resistance to cilostazol. Conclusions: We designed a method to monitor the effect of cilostazol using in vitro incubation with PGE1.

Cilostazol Induces Apoptosis and Inhibits Proliferation of Hepatocellular Carcinoma Cells by Activating AMPK

심경화,Mi-Sun Shu,김소영,김종연,최보현,이윤주 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.5

Hepatocellular carcinoma (HCC) is the most common primary liver cancer and one of the leading causes of cancer-related death. Cilostazol, an antiplatelet drug, elicits anticancer effects on human squamous cell carcinoma and colon cancer cells. We previously reported that cilostazol protects normal mature hepatocytes from alcohol-induced apoptotic cell death. In addition, cilostazol stimulates liver regeneration after hepatectomy. Therefore, this study evaluated whether cilostazol elicits pro- or antiproliferative effects on HCC using Hep3B and SK-Hep1 cells. Cilostazol inhibited proliferation of HCC cells by inducing apoptosis. Additionally, cilostazol induced G0/G1 cell cycle arrest and decreased expression of cyclin D1 and proliferating cell nuclear antigen. Activation of AMPactivated protein kinase (AMPK) and inhibition of extracellular signal-regulated kinase (ERK) and AKT signaling were associated with the anti-proliferative effect of cilostazol. LY294002 and PD98059, inhibitors of AKT and ERK, respectively, enhanced the anti-proliferative effect of cilostazol. By contrast, inhibition of AMPK using compound C or AMPK-targeting siRNA abolished the anti-proliferative effect of cilostazol. Moreover, AMPK inhibition reversed the down-regulation of AKT/EKR induced by cilostazol, indicating negative cross-talk between AMPK and AKT/ERK. These findings provide evidence that cilostazol exerts anti-tumor activity in HCC by counter-regulating AMPK and AKT/ERK signaling. Taken together, our findings suggest that cilostazol may provide clinical benefits in HCC patients by selectively targeting HCC cells without interfering with liver function.

관상동맥내 스텐트 시술시 Aspirin과 Cilostazol 2개월 병합요법

윤명호,탁승제,염철훈,장혁재,최소연,유상용,안성균,고종훈,신준한,김한수,최병일 대한순환기학회 2000 Korean Circulation Journal Vol.30 No.8

Cilostazol attenuates murine hepatic ischemia and reperfusion injury via heme oxygenase-dependent activation of mitochondrial biogenesis

Joe, Yeonsoo,Zheng, Min,Kim, Hyo Jeong,Uddin, Md. Jamal,Kim, Seul-Ki,Chen, Yingqing,Park, Jeongmin,Cho, Gyeong Jae,Ryter, Stefan W.,Chung, Hun Taeg American Physiological Society 2015 American journal of physiology, Gastrointestinal a Vol.309 No.1

<P>Hepatic ischemia-reperfusion (I/R) can cause hepatocellular injury associated with the inflammatory response and mitochondrial dysfunction. We studied the protective effects of the phosphodiesterase inhibitor cilostazol in hepatic I/R and the roles of mitochondria and the Nrf2/heme oxygenase-1 (HO-1) system. Wild-type, <I>Hmox1</I><SUP><I>−/−</I></SUP>, or <I>Nrf2</I><SUP><I>−/−</I></SUP> mice were subjected to hepatic I/R in the absence or presence of cilostazol followed by measurements of liver injury. Primary hepatocytes were subjected to cilostazol with the HO-1 inhibitor ZnPP, or Nrf2-specific siRNA, followed by assessment of mitochondrial biogenesis. Preconditioning with cilostazol prior to hepatic I/R protected against hepatocellular injury and mitochondrial dysfunction. Cilostazol reduced the serum levels of alanine aminotransferase, TNF-α, and liver myeloperoxidase content relative to control I/R-treated mice. In primary hepatocytes, cilostazol increased the expression of HO-1, and markers of mitochondrial biogenesis, PGC-1α, NRF-1, and TFAM, induced the mitochondrial proteins COX III and COX IV and increased mtDNA and mitochondria content. Pretreatment of primary hepatocytes with ZnPP inhibited cilostazol-induced PGC-1α, NRF-1, and TFAM mRNA expression and reduced mtDNA and mitochondria content. Genetic silencing of Nrf2 prevented the induction of HO-1 and mitochondrial biogenesis by cilostazol in HepG2 cells. Cilostazol induced hepatic HO-1 production and mitochondrial biogenesis in wild-type mice, but not in <I>Hmox1</I><SUP><I>−/−</I></SUP> or <I>Nrf2</I><SUP><I>−/−</I></SUP> mice, and failed to protect against liver injury in <I>Nrf2</I><SUP><I>−/−</I></SUP> mice. These results suggest that I/R injury can impair hepatic mitochondrial function, which can be reversed by cilostazol treatment. These results also suggest that cilostazol-induced mitochondrial biogenesis was mediated by an Nrf-2- and HO-1-dependent pathway.</P>

Cilostazol inhibits HMGB1 release in LPS-activated RAW 264.7 cells and increases the survival of septic mice

Pergamon Press 2015 Thrombosis research Vol.136 No.2

Introduction: Inflammation and coagulation play important roles in the pathogenesis of sepsis. Anticoagulants with anti-inflammatory action draw attention as therapeutic agent in sepsis. Objective: Whether cilostazol (6-[4-(1-cyclohexyl-1H-tetrazol-5-yl) butoxy]-3,4-dihydro-2-(1H)-quinolinone), anticoagulant, protects mice against sepsis and underlying mechanism(s) were investigated. Methods: Induction of heme oxygenase (HO)-1 protein, phosphorylation of 5' adenosine monophosphate-activated protein kinase (AMPK), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) luciferase activity, and release of high mobility group box 1 (HMGB1) were analyzed using signal inhibitors and transfection techniques. Survival and organ damage were compared in septic mice with and without cilostazol. Results: In RAW264.7 cells, cilostazol increased phosphorylation of AMPK which was followed by HO-1 induction. Lipopolysaccharide (LPS)-activated HMGB1 release was reduced by cilostazol which was reversed by both SB203580 and silencing of HO-1 or AMPK RNA. Interestingly, silencing AMPK reduced HO-1 expression, whereas silencing HO-1 did not affect p-AMPK by cilostazol. Both compound C and zinc protoporphyrin IX (ZnPPIX) antagonized inhibitory effect of HMGB1 by cilostazol. Cilostazol inhibited NF-κB luciferase activity which was antagonized by SB203580. Finally, the administration of cilostazol increased the survival of endotoxemic mice but failed to do so when co-treated with rHMGB1. Cilostazol reduced circulating HMGB1, plasminogen activator inhibitor-1 (PAI-1) levels, organ damages and protein expression of PAI-1 in lung tissues of CLP-septic mice, which were antagonized by ZnPPIX. Conclusion: These findings suggest that HMGB1 can be a target molecule of cilostazol by 1) AMPK activation, and 2) induction of HO-1 by p38 MAPK and AMPK. Therefore, cilostazol may be useful for treatment of sepsis.

자가혈관 동정맥루 생존율에 대한 Cilostazol의 효과

김정섭 ( Jung Sub Kim ),최문기 ( Mun Ki Choi ),최보경 ( Bo Kyung Choi ),이희선 ( Hee Sun Lee ),이나리아 ( Naria Lee ),손정민 ( Jung Min Son ),성은영 ( Eun Young Seong ),송상헌 ( Sang Heon Song ),이수봉 ( Soo Bong Lee ),곽임수 ( Ihm 대한신장학회 2010 Kidney Research and Clinical Practice Vol.29 No.4

Purpose: Vascular access failure is the most common reason for hospitalization among hemodialysis (HD) patients. Cilostazol, which has antiplatelet action and vasodialtory effects, significantly reduces the risk of restenosis after percutaneous coronary intervention in many patients. We conducted this study to evaluate the relationship between the use of antiplatelet agents, especially cilostazol, and arteriovenous fistula (AVF) patency in HD patients. Methods: A total of 241 patients underwent native AVF creation from January 2001 to December 2008. Among these patients, we selected 86 patients excluding 38 patients (15.8%) with primary technical failure, 49 patients without complete data and 68 patients used cilostazol less than 1 month. Demographic characteristics, medication history and fistula failure rate were collected and analyzed to elucidate the effect of cilostazol to native AVF. Results: From all groups, AVF failure occurred in 24 patients (27.9%). 28 patients received cilostazol (62.3 %) and mean duration of cilostazol therapy was 229.5±115.7 days. All patients were classified into two groups according to cilostazol (Cilostazol [n=28, 32.6%] vs. non-Cilostazol [n=58, 67.4%]. There was no statisticallly significant difference in failure rate between the two groups (32.1% vs. 25.9%, p=0.543). In diabetes group, patients who received statin have much lower AVF failure rate (0 % vs. 32.4%, p=0.024). Logistic regression analysis showed that female was independent risk factor for access failure (HR 5.549, CI 1.104-27.877, p=0.037). Conclusion: Cilostazol and other antiplatelet agent had a no significant association with AVF patency. Female was an independent risk factor for access failure.

약물방출 스텐트 시술 후 항 혈소판 삼제 복합치료의 효과

김동한 ( Dong Han Kim ),김준영 ( Joon Young Kim ),문승원 ( Seung Won Moon ),정종혁 ( Jong Hyuk Jung ),양혁승 ( Hyuk Seung Yang ),조장현 ( Jang Hyun Cho ),정명호 ( Myung Ho Jeong ) 대한내과학회 2008 대한내과학회지 Vol.74 No.4

목적: Cilostazol은 phosphodiesterase (PDE) III를 선택적으로 차단하는 항혈소판 제제로 혈관 내막증식 억제 작용을 함께 가지고 있다. 최근의 임상연구에서 관상동맥 내 BMS 삽입 후 cilostazol을 포함한 삼제 병합치료(aspirin+clopido-grel+cilostazol)를 하였을 경우 표준치료(aspirin+clopidogrel+placebo)에 비하여 유의한 재협착률 감소를 나타내었다. 본 연구는 DES 삽입 후 저용량의 cilostazol을 포함한 6개월 간의 항혈소판 삼제 병합요법의 효과를 알아보고자 하였다. 방법: 2004년 6월부터 2006년 1월까지 순천 성가롤로 병원에서 관상동맥 내 DES 삽입술을 시행 받은 환자를 대상으로 6개월간 저용량의 cilostazol (50 mg/BID) 복용 여부에 따라 I군(aspirin+clopidogrel+cilostazol)과 II군(aspirin+clopidogrel)으로 나누어 추적 관상동맥 조영술 및 임상결과를 전향적으로 비교하였다. 결과: 대상 환자는 109명이었으며 I군은 56명(67.3±10.8세, 남자 37명), II군은 53명(67.6±10.5세, 남자 26명)이었다. 대상 환자 중 80명(74%)에서 6개월 후 추적 관상동맥 조영술을 시행하였고, 임상 추적관찰은 모든 환자에서 실시되었다. 추적 관상동맥 조영술상 양 군 간에 최소 혈관 내경(I군: 2.25±0.63 mm, II군: 2.30±0.56 mm, p=0.742), 후기 손실(I군: 0.47±0.47 mm, II군: 0.52±0.53 mm, p=0.747)과 재협착률 (I군: 7.2%; II군: 5.6%, p=0.611)의 유의한 차이는 없었다. 또한 임상 추적 결과상 스텐트 혈전증은 일어나지 않았고, 양군간에 심장관련 사고나 출혈성 합병증 발생률의 유의한 차이는 없었다. 결론: DES 삽입 후 저용량의 cilostazol을 포함한 6개월 간의 항혈소판 삼제 병합치료는 출혈성 합병증을 증가시키지 않고 비교적 안전하게 사용할 수 있지만 보다 큰 최소혈관 내경을 얻고 재협착률을 감소시키는데 효과가 없었다. Background/Aims: A recent clinical trial demonstrated that triple anti-platelet therapy resulted in significantly larger minimal luminal diameter and lower restenosis rate compared with conventional therapy after bare-metal stent (BMS) implantation. However, it is uncertain that this result will be repeated after drug-eluting stent (DES) implantation, especially with low-dose cilostazol therapy. Thus, we performed a prospective, randomized study to evaluate the effectiveness of long-term triple therapy with low-dose cilostazol after DES implantation. Methods: We analyzed 109 patients (132 lesion) prospectively, who underwent successful coronary DES implantation. The patients were divided into two groups according to combined anti-platelet regimen: triple combination of aspirin, clopidogrel, and low-dose cilostazol (50 mg/bid) (Group I, n=56) or dual combination of aspirin and clopidogrel (Group II, n=53) for 6 months. The minimal luminal diameter and binary restenosis rate were compared at 6-month follow-up by coronary angiogram. The rates of stent thrombosis, major adverse cardiac events (MACE), and bleeding complication were also analyzed. Results: The baseline clinical and angiographic characteristics were not different between the two groups. Angiographic follow-up was performed in 80 patients (109 lesions, 74%). The minimal luminal diameter at 6 month was 2.25±0.63 mm in group I and 2.30±0.56 mm in group II (p=0.742). Restenosis occurred in 4 patients (7.2%) in group I and 3 patients (5.6%) in group II (p=0.611). There were no differences in the rates of stent thrombosis, MACE, or bleeding complications between the two groups. Conclusions: Long-term triple anti-platelet therapy with low-dose cilostazol after DES implantation was not effective in obtaining larger minimal luminal diameter or reducing restenosis rate, but it was used safely without increasing bleeding complication.(Korean J Med 74:368-375, 2008)

Cilostazol Decreases Ethanol-Mediated TNFalpha Expression in RAW264.7 Murine Macrophage and in Liver from Binge Drinking Mice

Lee, Youn-Ju,Eun, Jong-Ryeol The Korean Society of Pharmacology 2012 The Korean Journal of Physiology & Pharmacology Vol.16 No.2

Alcoholic hepatitis is a leading cause of liver failure in which the increased production of tumor necrosis factor ${\alpha}$ (TNF${\alpha}$) plays a critical role in progression of alcoholic liver disease. In the present study, we investigated the effects of cilostazol, a selective inhibitor of type III phosphodiesterase on ethanol-mediated TNF${\alpha}$ production in vitro and $in$ $vivo$, and the effect of cilostazol was compared with that of pentoxifylline, which is currently used in clinical trial. RAW264.7 murine macrophages were pretreated with ethanol in the presence or absence of cilostazol then, stimulated with lipopolysacchride (LPS). Cilostazol significantly suppressed the level of LPS-stimulated TNF${\alpha}$ mRNA and protein with a similar degree to that by pentoxifylline. Cilostazol increased the basal AMP- activated protein kinase (AMPK) activity as well as normalized the decreased AMPK by LPS. AICAR, an AMPK activator and db-cAMP also significantly decreased TNF${\alpha}$ production in RAW264.7 cells, but cilostazol did not affect the levels of intracellular cAMP and reactive oxygen species (ROS) production. The $in$ $vivo$ effect of cilostazol was examined using ethanol binge drinking (6 g/kg) mice model. TNF${\alpha}$ mRNA and protein decreased in liver from ethanol gavaged mice compared to that from control mice. Pretreatment of mice with cilostazol or pentoxifylline further reduced the TNF${\alpha}$ production in liver. These results demonstrated that cilostazol effectively decrease the ethanol-mediated TNF${\alpha}$ production both in murine macrophage and in liver from binge drinking mice and AMPK may be responsible for the inhibition of TNF${\alpha}$ production by cilostazol.