P2 receptor; CYP2; CYP3; adenosine diphosphate/ADP receptor

噻氯匹啶是一种有效的血小板聚集抑制剂。它是一种代谢为活性形式的前药，可阻断参与GPIIb/IIIa受体激活导致血小板聚集的ADP受体。噻氯匹啶以Ticlid品牌销售，适用于不能服用阿司匹林或阿司匹林对预防血栓性卒中无效的患者。美国食品药品监督管理局的标签包括中性粒细胞减少症、再生障碍性贫血、血栓性血小板减少性紫癜和粒细胞缺乏症的黑框警告，因此有必要在服用噻氯匹定时监测患者的白细胞和血小板。

---

噻氯匹定显示出针对人 CD39 的表观 Ki,app 值为 14 µM 的活性[1]。 Ticlopidine 的 Ki 值为 127±12 µM，可抑制 COS-7 细胞中表达的重组人 CD39[1]。噻氯匹定（30 和 150 µM）对培养最初几天的生长速率有影响，但其影响在随后的几天内减弱[4]。

---

研究了不同浓度（150、30、6微M）的噻氯匹啶对培养的人脐带静脉内皮细胞的影响。噻氯匹啶影响内皮细胞与人工基质的初始附着，并对内皮细胞生长速率具有抑制作用，这与培养基中化学物质的浓度有关。这些效应与免疫荧光所证明的细胞内外纤维连接蛋白的显著减少有关。该药物似乎会干扰细胞内颗粒形成纤维连接蛋白丝。纤维连接蛋白可用性的降低可能会影响血小板与内皮下的粘附以及内皮细胞的修复，进而影响出血时间。抑制细胞增殖及其对血管壁厚度的可能影响应被视为该物质的额外作用机制[4]。

---

CYP2C9和3A4的抑制作用[3]
噻氯匹啶对CYP2C9和3A4活性的抑制作用如图1所示。噻氯匹啶抑制CYP2C9和3A4，IC50值分别为26.0和32.3μmol/L。

---

罗丹明-123保留试验[3]
噻氯匹啶对罗丹明123在MCF-7和MCF-7/ADR细胞中的细胞积累的影响如图2所示。与缺乏P-gp的MCF-7细胞相比，过表达P-gp的MC-7/ADR细胞中罗丹明123的积累减少。在1-30μmol/L的噻氯匹定浓度范围内，两种条件下罗丹明123的相对细胞摄取量相当。

噻氯匹定与氯沙坦按 10 mg/kg 剂量口服时，可有效阻止氯沙坦在肠道和/或肝脏中的代谢。该组合观察到的 AUC 显着上升（65.0%）表明了这一点[3]。

---

噻氯匹定（10mg/kg）显著增加了口服氯沙坦（9mg/kg）的血浆浓度-时间曲线下面积（AUC）和峰值血浆浓度（C（max）），以及活性代谢产物EXP-3174的AUC。噻氯匹定（10mg/kg）不会显著改变静脉注射氯沙坦（3mg/kg）的药代动力学。噻氯匹啶抑制IC的CYP2C9和3A4₅₀ 分别为26.0和32.3μmol/L。罗丹明123的相对细胞摄取没有变化。 结论：噻氯匹定（10mg/kg）显著增加氯沙坦（9mg/kg）的AUC可能是由于抑制了CYP2C9-和3A4介导的氯沙坦在小肠和/或肝脏中的代谢。噻氯匹啶对小肠P-gp的抑制和氯沙坦肾排泄的减少不太可能是致病因素。[3]

---

噻氯匹定对口服氯沙坦药代动力学的影响[3]
大鼠口服氯沙坦（9mg/kg），同时口服和不口服噻氯匹定，剂量分别为4和10mg/kg。氯沙坦的平均动脉血浆浓度-时间曲线如图3所示。氯沙坦的相关药代动力学参数如表1所示。氯沙坦吸收快；所有研究的大鼠在第一个血液取样时间点（5分钟）血浆中检测到氯沙坦，tmax快速（1-2h）（图3）。口服氯沙坦和10mg/kg噻氯匹定后，氯沙坦的AUC和Cmax明显高于对照组大鼠（分别为65.0%和49.4%）。氯沙坦的F值为64.7%。

---

噻氯匹定对活性代谢产物EXP-3174药代动力学的影响[3]
大鼠口服氯沙坦（9mg/kg），同时口服和不口服噻氯匹定，剂量分别为4和10mg/kg。EXP-3174的平均动脉血浆浓度-时间曲线如图4所示。氯沙坦的相关药代动力学参数如表2所示。口服氯沙坦和10mg/kg噻氯匹定后，EXP-3174的AUC和Cmax明显大于对照组大鼠（分别增加41.8%和36.8%）。与对照组大鼠相比，AUCEXP-3174/AUClosartan的AUC比值没有显著降低（P>0.05）。

---

噻氯匹定对静脉注射氯沙坦药代动力学的影响[3]
图5显示了在有或没有噻氯匹定（4和10mg/kg）的情况下，大鼠静脉注射氯沙坦（3mg/kg）后氯沙坦的平均动脉血浆浓度-时间曲线。相应的药代动力学参数如表3所示。 噻氯匹定治疗增加了氯沙坦的AUC，但与对照组相比没有统计学意义。氯沙坦的t1/2也延长，但这种增加并不显著。与口服氯沙坦相比，静脉注射氯沙坦的药代动力学不受噻氯匹定同时使用的影响。因此，在噻氯匹定存在的情况下，口服生物利用度提高，而静脉注射氯沙坦的药代动力学没有显著变化。这一发现可能是由于抑制了CYP3A介导的氯沙坦在小肠和/或肝脏中的代谢，而不是噻氯匹定减少了氯沙坦在肾脏中的清除。

盐酸噻氯匹啶是噻氯匹定的盐酸盐形式，噻氯匹定是一种具有抗凝血特性的噻吩并吡啶衍生物。盐酸噻氯匹啶通过与糖蛋白（GP）IIb/IIIA复合物结合，不可逆地抑制二磷酸腺苷（ADP）诱导的血小板纤维蛋白原结合，糖蛋白是ADP激活的两种嘌呤能受体之一。受体激活的抑制会导致腺苷酸环化酶的抑制，导致环磷酸腺苷水平降低，从而干扰血小板膜功能和随后的血小板-血小板相互作用、血小板颗粒成分的释放和出血时间的延长。

细胞增殖测定[4]
细胞类型： 人内皮细胞
测试浓度： 30 和 150 µM
孵育时间： 2, 6; 10 天
实验结果：与对照组相比，处理过的细胞生长较慢，这种效应与培养基中噻氯匹定的浓度相关。

---

罗丹明-123保留试验[3]
使用MCF-7/ADR，一种对阿霉素耐药的人癌症细胞系，并以105个细胞的密度接种在24孔板上。在80%融合时，将细胞在无FBS的DMEM中孵育18小时。然后将培养基换成Hanks的平衡盐溶液，在37°C下孵育30分钟。在噻氯匹啶（1、3、10和30μmol/L）存在下，用20μmol/L罗丹明123孵育细胞90分钟后，完全去除培养基。然后用冰冷的磷酸盐缓冲液（pH 7.0）洗涤细胞三次，并在裂解缓冲液中裂解。分别使用480和540nm的激发和发射波长在细胞裂解物中测量罗丹明-123荧光。荧光值被归一化为每个样品的总蛋白质含量，并与对照值相比以比率表示。维拉帕米（100μmol/L）作为阳性对照。

Animal/Disease Models: Male SD (Sprague-Dawley) rats ( 7-8 weeks old, weighing 270-300 g)[3]
Doses: 4 or 10 mg/kg
Route of Administration: Orally administered 30 min before oral administration of losartan.
Experimental Results: The AUC and Cmax of Losartan after oral administration with Losartan and 10 mg/kg Ticlopidine were Dramatically greater (by 65.0% and 49.4%, respectively) than those of control rats.

---

Oral and intravenous administrations of losartan and Ticlopidine [3]
Losartan was orally or intravenously administrated, whereas ticlopidine was administrated orally. Oral losartan and ticlopidine were dissolved in distilled water (5 and 4 mL, respectively). Intravenous losartan was dissolved in 0.9% NaCl solution (4 mL). The rats were randomly divided into six groups (n=6 each): oral losartan (9 mg/kg) without and with oral ticlopidine (4 or 10 mg/kg); intravenous losartan (3 mg/kg) without and with oral ticlopidine (4 or 10 mg/kg). The rats were fasted for at least 24 h prior to the beginning of the experiments. Each animal was anaesthetized with light ether and the right femoral artery (for blood sampling) or vein (for iv administration of losartan) was cannulated with a polyethylene tube (SP45, ID 0.58 mm, OD 0.96 mm). Ticlopidine was orally administered 30 min before oral administration of losartan. Oral losartan and ticlopidine were administered using a gastric gavage tube. A blood sample (approximately 0.5 mL) was collected into a heparinized tube at 0 (control), 0.017 (end of the infusion), 0.1, 0.25, 0.5, 1, 2, 4, 8, and 12 h after iv administration, and 0, 0.1, 0.25, 0.5, 1, 2, 4, 6, 8, and 12 h after oral administration. Approximately 1 mL of whole blood collected from untreated rats was infused via the femoral artery at 0.25, 1, and 4 h to replace the blood loss due to blood sampling. Each blood sample was centrifuged at 16 810×g for 5 min, and 200 μL of each plasma sample was stored at -40 °C until HPLC analysis of losartan and EXP-3174 was performed.

Absorption, Distribution and Excretion
Absorption is greater than 80%. Food increases absorption by approximately 20%.
Ticlopidine is eliminated mostly in the urine (60%) and somewhat in the feces (23%).
The volume of distribution was not quantified.
Ticlopidine clearance was not quantified, but clearance decreases with age.

---

Metabolism / Metabolites
Ticlopidine is metabolized extensively by the liver with only trace amounts of intact drug detected. At least 20 metabolites have been identified.
Ticlopidine has known human metabolites that include Ticlopidine S-oxide and Thienodihydropyridinium.

---

Biological Half-Life
Half-life following a single 250-mg dose is approximately 7.9 hours in subjects 20 to 43 years of age and 12.6 hours in subjects 65 to 76 years of age. With repeated dosing (250 mg twice a day), half-life is about 4 days in subjects 20 to 43 years of age and about 5 days in subjects 65 to 76 years of age.

Hepatotoxicity
Ticlopidine has been associated with serum enzyme elevations in approximately 4% of patients during therapy. These elevations are usually mild, asymptomatic and rarely require dose modification or stopping. Ticlopidine has also been associated with clinically apparent, acute liver injury. While these reactions are rare, more than 50 instances have been reported in the literature and some have been severe. The onset of symptoms is typically within 6 weeks (range 1 to 24 weeks) and marked by onset with fatigue, jaundice and itching. The usual pattern of liver enzyme elevations is cholestatic (~75%), but cases with mixed or hepatocellular enzyme elevations have also been described. Immunoallergic features such as fever, rash and eosinophilia can occur but are not common and, if present, are usually mild. Autoantibody formation is rare. Liver biopsy usually shows cholestatic hepatitis with mixed cellular infiltrates. Most cases are self-limited with recovery within 1 to 3 months, but isolated cases of prolonged jaundice or liver test abnormalities have been described, including at least one case of probable vanishing bile duct syndrome that eventually required liver transplantation. Ticlopidine therapy has also been associated with aplastic anemia and thrombotic thrombocytopenic purpura (TTP) that can be severe and lead to death; these patients may also have accompanying cholestatic liver injury.

---

Protein Binding
Binds reversibly (98%) to plasma proteins, mainly to serum albumin and lipoproteins. The binding to albumin and lipoproteins is nonsaturable over a wide concentration range. Ticlopidine also binds to alpha-1 acid glycoprotein (about 15% or less).

[1]. 2-Substituted thienotetrahydropyridine derivatives: Allosteric ectonucleotidase inhibitors. Arch Pharm (Weinheim). 2021 Dec;354(12):e2100300.

[2]. Ticlopidine-Induced Cholestatic Inflammatory Hepatitis: New Insights into Pathogenetic Mechanisms of Drug-Related Hepatotoxicity.

[3]. Effects of ticlopidine on pharmacokinetics of losartan and its main metabolite EXP-3174 in rats. Acta Pharmacol Sin. 2011 Jul;32(7):967-72.

[4]. The effect of Ticlopidine on human endothelial cells in culture. Thromb Res. 1984 Feb 1;33(3):323-32.

Ticlopidine is a thienopyridine that is 4,5,6,7-tetrahydrothieno[3,2-c]pyridine in which the hydrogen attached to the nitrogen is replaced by an o-chlorobenzyl group. It has a role as a fibrin modulating drug, a hematologic agent, an anticoagulant, a platelet aggregation inhibitor and a P2Y12 receptor antagonist. It is a thienopyridine and a member of monochlorobenzenes.
Ticlopidine is an effective inhibitor of platelet aggregation. It is a prodrug that is metabolised to an active form, which blocks the ADP receptor that is involved in GPIIb/IIIa receptor activation leading to platelet aggregation. Ticlopidine is marketed under the brand name Ticlid and is indicated for patients who cannot take aspirin or in whom aspirin has not worked to prevent a thrombotic stroke. The FDA label includes a black-box warning of neutropenia, aplastic anemia, thrombotic thrombocytopenia purpura, and agranulocytosis, so it is necessary to monitor patients' WBC and platelets when they are taking ticlopidine.
Ticlopidine is a Platelet Aggregation Inhibitor. The physiologic effect of ticlopidine is by means of Decreased Platelet Aggregation.
Ticlopidine is an inhibitor of platelet aggregation that is used to decrease the risk of stroke in patients known to have atherosclerosis. Ticlopidine is associated with a low rate of serum enzyme elevations during treatment and has been linked to rare instances of idiosyncratic, clinically apparent acute liver injury.
Ticlopidine is a thienopyridine derivative with anticoagulant activity. Ticlopidine inhibits adenosine-diphosphate (ADP) binding to its platelet receptor. This prevents ADP activation and inhibits platelet expression of the glycoprotein (GP) IIb/IIIA receptors, binding of fibrinogen to platelet glycoprotein GP IIb-IIIa and platelet-platelet interaction. This results in increased bleeding time.
An effective inhibitor of platelet aggregation commonly used in the placement of STENTS in CORONARY ARTERIES.
See also: Ticlopidine Hydrochloride (has salt form).

---

Drug Indication
Used in patients, who have had a stroke or stroke precursors and who cannot take aspirin or aspirin has not worked, to try to prevent another thrombotic stroke.
FDA Label

---

Mechanism of Action
The active metabolite of ticlopidine prevents binding of adenosine diphosphate (ADP) to its platelet receptor, impairing the ADP-mediated activation of the glycoprotein GPIIb/IIIa complex. It is proposed that the inhibition involves a defect in the mobilization from the storage sites of the platelet granules to the outer membrane. No direct interference occurs with the GPIIb/IIIa receptor. As the glycoprotein GPIIb/IIIa complex is the major receptor for fibrinogen, its impaired activation prevents fibrinogen binding to platelets and inhibits platelet aggregation. By blocking the amplification of platelet activation by released ADP, platelet aggregation induced by agonists other than ADP is also inhibited by the active metabolite of ticlopidine.

---

Pharmacodynamics
Ticlopidine is a prodrug that is metabolised to an as yet undetermined metabolite that acts as a platelet aggregation inhibitor. Inhibition of platelet aggregation causes a prolongation of bleeding time. In its prodrug form, ticlopidine has no significant in vitro activity at the concentrations attained in vivo.

---

Aim: Losartan and antiplatelet agent ticlopidine can be prescribed concomitantly for prevention or therapy of cardiovascular diseases. Hence, the effects of ticlopidine on the pharmacokinetics of losartan and its active metabolite EXP-3174 were evaluated in rats. Methods: Ticlopidine (4 or 10 mg/kg po) was administered 30 min before administration of losartan (9 mg/kg po or 3 mg/kg iv). The activity of human CYP2C9 and 3A4 were measured using the CYP inhibition assay kit. The activity of P-gp was evaluated using rhodamine-123 retention assay in MCF-7/ADR cells. [3]

---

The significant increase in the AUC of losartan after oral administration with losartan and ticlopidine (10 mg/kg) may be attributable to the inhibition of CYP2C and 3A subfamilies-mediated losartan metabolism in the small intestine and/or in the liver. The inhibition of P-gp in the small intestine and the reduction of renal elimination of losartan by ticlopidine are unlikely to be causal factors.[3]

C14H14CLNS

263.79

263.053

55142-85-3

Ticlopidine hydrochloride;53885-35-1;Ticlopidine-d4;1246817-49-1

5472

Light yellow to yellow oil

1.3±0.1 g/cm3

367.3±37.0 °C at 760 mmHg

approx. 1 189°C

175.9±26.5 °C

0.0±0.8 mmHg at 25°C

1.638

3.77

31.48

0

2

2

17

261

0

PHWBOXQYWZNQIN-UHFFFAOYSA-N

InChI=1S/C14H14ClNS/c15-13-4-2-1-3-11(13)9-16-7-5-14-12(10-16)6-8-17-14/h1-4,6,8H,5,7,9-10H2

5-[(2-chlorophenyl)methyl]-6,7-dihydro-4H-thieno[3,2-c]pyridine

5-(2-Chlorobenzyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine; Ticlopidina; Ticlopidinum; Ticlopidin-Puren; PCR 5332; Ticlopidinum [INN-Latin];

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 本产品在运输和储存过程中需避光。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ≥ 100 mg/mL (379.09 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (9.48 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

---

配方 2 中的溶解度: ≥ 2.5 mg/mL (9.48 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

---

请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。