西洛他唑与羟丙基-β-环糊精包合的热力学研究

目的：研究西洛他唑（CLTZ）与羟丙基-β-环糊精（HP-β-CD）在混合溶液中的包合作用及其包合过程中热力学参数的变化。方法：运用 RP-HPLC 法测定 CLTZ 与 HP-β-CD 在混合溶液中包合的平衡常数（Kf）,计算与分析其热力学的参数变化。结果：CLTZ 与 HP-β-CD 可形成 mol∶mol（1∶1）包合物,包合过程可自发进行（ΔG＜0）,且为放热反应（ΔH＜0）,也是熵减过程（ΔS＜0）。结论：CLTZ 与 HP-β-CD 可形成 mol∶mol（1∶1）可溶性包合物,从而增加其溶解度,且较低的温度有利于包合物的形成和稳定。     

Cilostazol suppresses β‐amyloid production by activating a disintegrin and metalloproteinase 10 via the upregulation of SIRT1‐coupled retinoic acid receptor‐β

The accumulation of plaques of β‐amyloid (Aβ) peptides, a hallmark of Alzheimer's disease, results from the sequential cleavage of amyloid precursor protein (APP) by activation of β‐ and γ‐secretases. However, the production of Aβ can be avoided by alternate cleavage of APP by α‐and γ‐secretases. We hypothesized that cilostazol attenuates Aβ production by increasing a disintegrin and metalloproteinase 10 (ADAM10)/α‐secretase activity via SIRT1‐coupled retinoic acid receptor‐β (RARβ) activation in N2a cells expressing human APP Swedish mutation (N2aSwe). To evoke endogenous Aβ overproduction, the culture medium was switched from medium containing 10% fetal bovine serum (FBS) to medium containing 1% FBS, and cells were cultured for 3～24 hr. After depletion of FBS in media, N2aSwe cells showed increased accumulations of full‐length APP (FL‐APP) and Aβ in a time‐dependent manner (3–24 hr) in association with decreased ADAM10 protein expression. When pretreated with cilostazol (10–30 μM), FL‐APP and Aβ levels were significantly reduced, and ADAM10 and α‐secretase activities were restored. Furthermore, the effect of cilostazol on ADAM10 expression was antagonized by pretreating Rp‐cAMPS and sirtinol and by SIRT1‐gene silencing. In the N2aSwe cells overexpressing the SIRT1 gene, ADAM10, and sAPPα levels were significantly elevated. In addition, like all‐trans retinoic acid, cilostazol enhanced the protein expressions of RARβ and ADAM10, and the cilostazol‐stimulated ADAM10 elevation was significantly attenuated by LE135 (a RARβ inhibitor), sirtinol, and RARβ‐gene silencing. In conclusion, cilostazol suppresses the accumulations of FL‐APP and Aβ by activating ADAM10 via the upregulation of SIRT1‐coupled RARβ. © 2014 Wiley Periodicals, Inc.     

Indomethacin-induced small intestinal injury is ameliorated by cilostazol,a specific PDE-3 inhibitor

Abstract

Background. Neutrophil migration, one of the major factors predisposing to nonsteroidal anti-inflammatory drugs (NSAIDs)-induced intestinal lesions, consists of several steps, including interaction with P-selectin from platelets. Cilostazol, a specific phosphodiesterase (PDE)-3 inhibitor, suppresses the expression of P-selectin from platelets and reduces interaction between platelets and leukocytes, leading to inflammatory amelioration in several disease models. We tried to clarify the therapeutic effectiveness of cilostazol for NSAID-induced small intestinal lesions. Subjects and methods. 1) Anti-PSGL-1 antibody (2 mg/kg) or cilostazol (100 mg/kg) was administered to mice one hour before Indomethacin (IND, 2.5 mg/kg) administration for 4 days to evaluate small intestinal lesions. 2) IND-induced migratory behaviors of neutrophils and platelets were evaluated in intestinal vessels by an intravital microscopy. Results. i) IND induced small intestinal lesions with an increase in MPO activity. Anti-PSGL-1 antibody and cilostazol ameliorated intestinal lesions along with suppression of MPO activity. ii) Intravital microscopy revealed that administration of IND increased migration of platelet-bearing neutrophils. Cilostazol treatment ameliorated neutrophil migration by blocking interaction between platelets and neutrophils. Conclusion. Our results suggest that enhanced platelets-bearing neutrophil migration is critically involved in the pathogenesis of IND-induced small intestinal lesions and suggest a potential application of cilostazol for prevention of NSAID-induced small intestinal lesions.     

西洛他唑对脑缺血后神经元损伤的保护作用

目的：观察西洛他唑对小鼠持续性局灶性脑缺血后神经元损伤的剂量依赖性保护作用。方法：以大脑中动脉阻塞诱导小鼠持续性局灶性脑缺血。缺血前30min腹腔注射西洛他唑（3～30mg/kg）和普鲁司特（0.1mg/kg）。观察药物对缺血后神经元形态、密度的影响。结果：脑缺血损伤后,神经元密度降低,变性神经元密度增加。西洛他唑（3～10mg/kg）和普鲁司特能明显增加缺血侧存活神经元密度,减少变性神经元密度。结论：西洛他唑对小鼠持续性局灶性脑缺血后神经元损伤有保护作用。     

Reevaluating the role of phosphodiesterase inhibitors in the treatment of cardiovascular disease

First developed for clinical use in the late 1980s, the phosphodiesterase inhibitors were found to increase the levels of the ubiquitous second messenger cyclic adenosine monophosphate and could effect changes in vascular tone, cardiac function, and other cellular events. After several early studies using high doses of phosphodiesterase inhibitors in patients with severe heart failure suggested adverse consequences, they fell out of favor. However, recent investigations of phosphodiesterase inhibitors in patients with intermittent claudication have demonstrated profound benefits. Furthermore, these agents have proven useful in prevention of cerebral infarction and coronary restenosis, and their use in the treatment of heart failure is being reevaluated. The reemergence of phosphodiesterase inhibitors can be attributed to a better understanding of dosing and drug-specific pharmacology, the use of concomitant medications, and a recognition of unique ancillary properties; however, their use still requires caution.     

前列地尔、硫辛酸联合西洛他唑治疗糖尿病周围神经病变疗效观察

目的:观察前列地尔、硫辛酸和西洛他唑3药联合使用对糖尿病周围神经病变(DPN)的疗效。方法:DPN患者76例,随机分为对照组和治疗组各38例,在糖尿病基础治疗基础上,对照组给予前列地尔和硫辛酸治疗,治疗组给予前列地尔、硫辛酸和西洛他唑3药联用。于治疗前和治疗4周后,检测并分析2组运动神经传导速度(MCV)、感觉神经传导速度(SCV)、运动神经复合肌动作电位(CMAP)波幅、CMAP潜伏期和感觉神经动作电位(SNAP)波幅的变化和神经病变主觉症状问卷(TSS)评分。结果:治疗前,2组TSS评分、神经传导速度、CMAP波幅、CMAP潜伏期和SNAP波幅无统计学差异(P>0.05);治疗后,2组TSS评分和CMAP潜伏期均低于治疗前,且治疗组低于对照组(P<0.01);2组MCV、CMAP波幅、SNAP波幅和SCV均较治疗前提高(P<0.01),且治疗组优于对照组(P<0.05)。结论:前列地尔、硫辛酸和西洛他唑3药联用治疗DPN疗效好于前列地尔、硫辛酸2药联用。     

Anti-platelet therapy: phosphodiesterase inhibitors

Inhibition of platelet aggregation can be achieved either by the blockade of membrane receptors or by interaction with intracellular signalling pathways. Cyclic adenosine 3',5'-monophosphate (cAMP) and cyclic guanosine 3',5'-monophosphate (cGMP) are two critical intracellular second messengers provided with strong inhibitory activity on fundamental platelet functions. Phosphodiesterases (PDEs), by catalysing the hydrolysis of cAMP and cGMP, limit the intracellular levels of cyclic nucleotides, thus regulating platelet function. The inhibition of PDEs may therefore exert a strong platelet inhibitory effect. Platelets possess three PDE isoforms (PDE2, PDE3 and PDE5), with different selectivity for cAMP and cGMP. Several nonselective or isoenzyme-selective PDE inhibitors have been developed, and some of them have entered clinical use as antiplatelet agents. This review focuses on the effect of PDE2, PDE3 and PDE5 inhibitors on platelet function and on the evidence for an antithrombotic action of some of them, and in particular of dipyridamole and cilostazol.     

Interaction analysis between genetic polymorphisms and pharmacodynamic effect in patients treated with adjunctive cilostazol to dual antiplatelet therapy: results of the ACCEL-TRIPLE (Accelerated Platelet Inhibition by Triple Antiplatelet Therapy According to Gene Polymorphism) study

AIMS

Although adjunctive cilostazol to dual antiplatelet therapy can reduce the risks of clinical events after percutaneous coronary intervention (PCI), whether genetic polymorphism can influence the pharmacodynamics of this regimen has not been evaluated.

METHODS

One hundred and twenty-seven patients treated with PCI and taking triple antiplatelet therapy (≥1 month) were enrolled. Platelet reactivity was assessed by conventional aggregometry and the VerifyNow P2Y12 assay. High on-treatment platelet reactivity (HPR) was defined as 5 µm ADP-induced maximal platelet reactivity (Agg_max) >46%. CYP3A5*3, CYP2C19*2/*3 and ABCB1 3435C > T were genotyped.

RESULTS

CYP3A5*3 and ABCB1 3435C > T variants did not affect the antiplatelet effect of triple antiplatelet therapy. For non-carriers, one and two carriers of the CYP2C19 loss-of-function (LOF) allele, Agg_max consecutively increased after the addition of 5 µm[mean (95% confidence intervals): 24.6% (20.8 to 28.5%) vs. 28.7% (25.4 to 32.0%) vs. 32.3% (25.8 to 38.7%), P = 0.062, respectively] and 20 µm ADP [34.2% (29.3 to 39.0%) vs. 41.7% (37.8 to 45.6%) vs. 44.9% (37.9 to 51.9%), P = 0.007, respectively]. Likewise, late platelet reactivity and P2Y12 reaction units proportionally changed according to the number of CYP2C19 LOF alleles. HPRs were observed in 9.2% of subjects: 6.3%, 7.4% and 20.0% with 0, 1 and 2 carriers of CYP2C19 LOF allele(s) (P = 0.099). In multivariate analysis, carriage of two CYP2C19 LOF alleles was a significant predictor for the prevalence of HPR (odds ratio 5.78, 95% CI 1.21, 27.78, P = 0.028).

CONCLUSION

Among PCI-treated patients, the effect of triple antiplatelet therapy is influenced by the CYP2C19 LOF allele. Its clinical benefit needs to be validated according to the CYP2C19 metabolic phenotype in future clinical trials. [Adjunctive Cilostazol Versus High Maintenance dose ClopidogrEL in Acute Myocardial Infarction Patients According to CYP2C19 Polymorphism (ACCEL-AMI-2C19), NCT00915733 and Adjunctive Cilostazol Versus High Maintenance-dose Clopidogrel According to Cytochrome 2C19 Polymorphism (ACCEL-2C19), NCT01012193].     

HPLC-MS/MS法测定人血浆中西洛他唑浓度

目的建立灵敏快速检测人血浆中西洛他唑浓度的HPLC-MS/MS方法。方法采用色谱柱为Thermo Hy-purity C18（150 mm×2.1 mm,5μm,USA）,流动相为乙腈（含0.1%甲酸）-甲醇=25∶75（v/v）;流速为0.2mL.min-1;ESI＋离子源,MRM进行离子方式监测。质谱参数：源电压3 500V,源温度100℃,去溶剂化温度350℃,锥孔气流速50 L.h^-1,去溶剂化气流速350 L.h^-1;血浆处理采用乙腈直接沉淀萃取方法。结果血浆中西洛他唑检测方法的线性范围为4.90-2 510 ng.mL-1,最低检出浓度为4.90 ng.mL-1,血浆中西洛他唑的方法回收率在80%-120%,日内RSD〈10%,日间RSD〈15%。结论本方法用于测定人血浆中西洛他唑的浓度,简单、快速、灵敏。