P2Y12 Receptor

坎格勒四钠是唯一有效的静脉直接电位二磷酸腺苷 (ADP) P2Y12 受体钳抗剂[1]。坎格勒四钠的 hP2Y12 受体 pKb 为 8.6-9.2[3]。

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Cangrelor是一种选择性、快速可逆的P2Y12血小板受体抑制剂，可直接阻断二磷酸腺苷（ADP）诱导的血小板活化和聚集，并在五分钟内达到90%的血小板抑制水平[1]。

坎格雷洛四钠 (10 mg/kg) 不仅显着降低 BLM 诱导的炎症细胞因子 (PF4、CD40 L 和 MPO) 的释放，而且降低纤维化肺和外周血管中血管、中性粒细胞损伤和中性粒粒细胞损伤细胞聚集体的增加BLM处理的血液[2]。

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研究人员报告称，坎格雷是一种非特异性GPR17拮抗剂，通过抑制小鼠巨噬细胞炎症，部分缓解了肺纤维化。Cangrelor也是一种众所周知的抗血小板药物。为了测试坎格雷是否部分通过抑制血小板来减轻肺纤维化，使用博莱霉素（BLM）诱导C57BL/6 J小鼠的肺纤维化。我们发现，坎格雷（10mg/kg）不仅显著降低了BLM诱导的炎性细胞因子（PF4、CD40L和MPO）的释放，还降低了BLP治疗小鼠纤维化肺和外周血中血小板、中性粒细胞和血小板-中性粒细胞聚集体的增加。此外，坎格雷降低了BLM治疗的小鼠肺部CD40和MPO双阳性中性粒细胞的数量以及CD40的表达水平。基于这些结果，得出结论，坎格雷减轻了BLM诱导的小鼠肺部炎症和肺纤维化，部分是通过抑制血小板活化，从而减少了CD40-CD40L相互作用介导的血小板和中性粒细胞粘附引起的中性粒细胞浸润。Cangrelor可能是治疗肺纤维化的潜在药物。[2]

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方法在野生型和P2ry12-/-基因缺陷型小鼠中诱导完全弗氏佐剂（CFA）诱导的慢性炎症性疼痛，并使用强效、直接作用和可逆的P2Y12受体拮抗剂PSB-0739和坎格雷。结果在急性炎症期，CFA诱导的P2ry12-/-小鼠机械性痛觉过敏显著降低，持续14天，后爪中性粒细胞髓过氧化物酶活性和肿瘤坏死因子（TNF）-α和CXCL1（KC）水平的升高也有所减轻。在第14天，P2ry12-/-小鼠白细胞介素（IL）-1β、IL-6、TNF-α和KC水平的升高减弱。PSB-0739和坎格雷逆转了野生型小鼠的痛觉过敏，但对P2ry12-/-小鼠没有影响，局部应用时PSB-0738也有效。选择性NaV1.8通道拮抗剂A-803467可预防局部和全身PSB-0739的作用，表明NaV1.8渠道参与了抗痛觉过敏作用。在第14天，抗小鼠CD41抗体的血小板耗竭降低了野生型小鼠的痛觉过敏并减弱了促炎细胞因子反应，但在P2ry12-/-小鼠中没有。结论总之，P2Y12受体调节CFA诱导的痛觉过敏和局部炎症反应，血小板P2Y12接收器在慢性炎症阶段参与了这些作用[3]。

Animals and reagents [2]
C57BL/6 J mice (male, 6–8 weeks, 22−25 g) were used. The mice are free access to water and food in air-conditioned rooms (23 °C, relative humidity 50 %) on a 12 h light / dark cycle. Four treatments were performed in these mice: sham-operated control (Con, n = 6), cangrelor (Cang, 10 mg/kg, n = 6), bleomycin + saline (BLM, 3 mg/kg, n = 6) and bleomycin + cangrelor (BLM + Cang 10 mg/kg, n = 6). Cangrelor and bleomycin (Hisun Pharmaceutical Co., Ltd., China) were stored at 4 °C and diluted in saline before use.

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Experimental procedure and cangrelor administration [2]
According to the previous report (Zhan et al., 2018, 2018; Tanaka et al., 2017), pulmonary fibrosis was induced by intratracheal administration of bleomycin (BLM, 3 mg/kg) in C57BL/6 J mice, and cangrelor (10 mg/kg) was administrated via subcutaneous injection. BLM was administrated on day 0, the treatment of cangrelor was started 2 days before BLM administration and lasted for 16 days (once per day). On day 14, the mice were sacrificed by cervical dislocation after the pulmonary resistance was determined. The bronchoalveolar lavage fluid (BALF) was collected from right lung, then the right lung tissues were stored at -80 °C for Western blotting analysis and quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The left lung tissues were fixed in 10 % formaldehyde for histological inspection. The blood and BALF were collected for flow cytometry and ELISA assay.

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Mice were treated with P2Y12R antagonists, or with their vehicle (sterile saline), intraperitoneally ([dichloro‐[[[(2R,3S,4R,5R)‐3,4‐dihydroxy‐5‐[6‐(2‐methylsulfanylethylamino)‐2‐(3,3,3‐trifluoropropylsulfanyl)purin‐9‐yl]oxolan‐2‐yl]methoxy‐hydroxyphosphoryl]‐oxyhydroxyphosphoryl]methyl]phosphonic‐acid, cangrelor , 3 mg kg−1; The Medicines Company, Parsippany, NJ, USA), intraplantarly or intrathecally (1‐amino‐4‐[4‐phenylamino‐3‐sulfophenylamino]‐9,10‐dioxo‐9,10‐dihydroanthracene‐2‐sulfonate, PSB‐0739, 0.3 mg kg−1, selective P2Y12R antagonist synthesized by Y. Baqi and C. E. Müller). on days 3, 4, 7, 10 and 14 after CFA injection. The doses were chosen on the basis of our previous experiments: the pKB values of PSB‐0739 and cangrelor at human P2Y12Rs (hP2Y12Rs) were 9.8 and 8.6, respectively, whereas, in the doses applied in the present study (PSB‐0739, 0.3 mg kg−1 intrathecally; cangrelor , 3 mg kg−1 intraperitoneally), they reversed acute inflammatory pain for up to 96 h. Taking into account that the approximate blood volume of a 25‐mg mice is 1700 μL, these doses correspond to 5 μm and 50 μm, indicating maximal target inhibition. As a reference compound, aspirin, an alternative platelet antagonist, was used at a low dose (2‐acetyloxybenzoic acid, 20 mg kg−1 intraperitoneally). The mechanonociceptive thresholds of hind paws were measured 15 min or 30 min after intrathecal/intraperitoneal or intraplantar injections, with the exception of day 3, when PWT measurements were performed before drug administration. 5‐(4‐chlorophenyl)‐N‐(3,5‐dimethoxyphenyl)‐2‐furancarboxamide (A‐803467, 30 mg kg−1), a potent and selective NaV1.8 sodium channel antagonist or its vehicle (polyethylene glycol and dimethyl sulfoxide [9 : 1]) was administered intraperitoneally 5 min before the respective PSB‐0739/saline injection. The dose of A‐803467 was chosen on the basis of a previous study, and a submaximal dose (30 mg kg−1 intraperitoneally) in the reduction of mechanical allodynia was selected to reveal any additive interactions between PSB‐0739 and A‐803467. In some experiments, paw edema was also volumetrically quantified by plethysmometry (7140; Ugo Basile). [3]

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Assessment of platelet CD62P levels by flow cytometry [3]
To investigate how P2Y12R antagonists and antiplatelet agents administered via different routes altered platelet activation, we measured ADP‐induced changes in platelet CD62P levels ex vivo, in platelet‐rich plasma (PRP) samples. Wild‐type mice were treated with PSB‐0739 (0.3 mg kg−1 intrathecally), cangrelor (3 mg kg−1 intraperitoneally), aspirin (20 mg kg−1 intraperitoneally), or their vehicle. Blood samples were taken directly from the vena cava of anesthetized mice 15 min or 30 min after the treatment. Apyrase (1 U mL−1) was added to the samples to prevent ADP receptor desensitization. After 10 min of centrifugation at 150 × g, PRP was collected. Platelet activation was induced by ADP (500 μm), and changes in platelet CD62P levels were assessed after 60 min of incubation. Platelets were stained with anti‐human/mouse CD62P antibody for 10 min. Samples were acquired with a BD FACSVerse machine, and analyzed with BD facsuite software. Changes in CD62P mean fluorescence intensity values were determined on CD42d‐positive platelets.

Absorption, Distribution and Excretion
Following IV administration of [3H] cangrelor, 58% of radioactivity was recovered in urine. The remaining 35% of radioactivity was in feces, presumably following biliary excretion.
In a study in healthy volunteers administration at a dose of 30 mcg/kg bolus plus 4 mcg/kg/min showed a volume of distribution of 3.9 L.
The mean clearance is about 43.2 L/h.
/MILK/ It is not known whether Kengreal is excreted in human milk.
Following IV administration of 3(H) Kengreal 58% of radioactivity was recovered in urine. The remaining 35% of radioactivity was in feces, presumably following biliary excretion. The average elimination half-life of Kengreal is about 3-6 minutes.
In a study in healthy volunteers, Kengreal administration at a dose of 30 ug/kg bolus plus 4 mcg/kg/min showed a volume of distribution of 3.9 L. Plasma protein binding of Kengreal is about 97-98%.

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Metabolism / Metabolites
Cangrelor is deactivated rapidly in the circulation by dephosphorylation to its primary metabolite, a nucleoside, which has negligible anti-platelet activity. Cangrelor's metabolism is independent of hepatic function and it does not interfere with other drugs metabolized by hepatic enzymes.
Kengreal is deactivated rapidly in the circulation by dephosphorylation to its primary metabolite, a nucleoside, which has negligible anti-platelet activity. Kengreal's metabolism is independent of hepatic function and it does not interfere with other drugs metabolized by hepatic enzymes.

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Biological Half-Life
The average elimination half-life of cangrelor is about 3-6 minutes.
Following IV administration of 3(H) Kengreal, ... elimination half-life of Kengreal is about 3-6 minutes.

Toxicity Summary
IDENTIFICATION AND USE: Cangrelor is a platelet aggregation inhibitor and purinergic P2Y receptor antagonist. HUMAN STUDIES: Cangrelor is a potent intravenous platelet P2Y12 receptor antagonist with rapid onset and offset of action. In patients undergoing percutaneous coronary interventions (PCI), compared with control, cangrelor (30 ug/kg bolus, followed immediately by a 4 ug/kg per minute infusion for 2-4 hr or until the conclusion of the index PCI, whichever was longer) reduces periprocedural thrombotic complications without an increase in major bleeding complications, although minor bleeding is increased. In a large clinical trial program of patients undergoing PCI, cangrelor overdosing was rare and not associated with an increase in bleeding complications, an observation that may be attributed to its very short-half life and rapid offset of action. Platelet P2Y12 receptor expression is significantly increased and the receptor is constitutively activated in patients with type 2 diabetes mellitus, which contributes to platelet hyperactivity and limits antiplatelet drug efficacy in type 2 diabetes mellitus. Cangrelor was non-mutagenic and non-clastogenic in genetic toxicology studies, including chromosome aberration assay in human peripheral lymphocytes. ANIMAL STUDIES: Cangrelor had no significant effect on male or female rats fertility treated for 28 days, or on early embryonic development. In embryo-fetal development studies in rats, cangrelor produced dose-related fetal growth retardation characterized by increased incidences of incomplete ossification and unossified hind limb metatarsals. In rabbits, cangrelor was associated with increased incidences of abortion and intrauterine losses, as well as fetal growth retardation. Cangrelor was non-mutagenic and non-clastogenic in genetic toxicology studies, including in vitro bacterial gene mutation assay, mouse lymphoma thymidine kinase assay, and in vivo bone marrow micronucleus assay in mice.

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Hepatotoxicity
In several large clinical trials, serum ALT elevations were no more frequent with cangrelor therapy than with placebo [9% vs 12%] or with comparator arms [6.6% vs 6.8%] and no cases of clinically apparent liver injury with jaundice were reported. In addition, since marketing and release, there have been no published reports of clinically apparent liver injury or jaundice associated with cangrelor therapy and hepatotoxicity is not mentioned in the product label.
Likelihood score: E (unlikely cause of clinically apparent liver injury).

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Protein Binding
about 97-98%.

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Interactions
BACKGROUND: Agents that act as antagonists at P2Y(12) ADP receptors on platelets are in use (clopidogrel), and in development for use (cangrelor and prasugrel), in patients with cardiovascular disease. Cangrelor is a direct-acting reversible antagonist being developed for short-term infusion; clopidogrel and prasugrel are oral prodrugs that provide irreversible inhibition via transient formation of active metabolites. At the cessation of cangrelor infusion, patients are likely to receive clopidogrel or prasugrel as a means of maintaining antiplatelet therapy. OBJECTIVES: To apply an experimental in vitro approach to investigate the possibility that cangrelor influences the ability of the active metabolites of clopidogrel and prasugrel to inhibit ADP-mediated platelet function. METHODS: The effects of cangrelor and the active metabolites of clopidogrel (C-AM) and prasugrel (P-AM) on platelet function were assessed by ADP-induced platelet P-selectin expression in whole blood. The method involved rapid removal of the antagonists by dilution, and measurement of residual platelet inhibition. RESULTS: Cangrelor, C-AM and P-AM markedly inhibited P-selectin expression. The effect of cangrelor, but not of C-AM and P-AM, was reversible following antagonist removal. Preincubation of blood with cangrelor prior to addition of C-AM or P-AM reduced the ability of metabolites to irreversibly antagonize P2Y(12). Irreversible inhibition was maintained when blood was preincubated with metabolites prior to cangrelor. CONCLUSIONS: Cangrelor influences the ability of the active metabolites of clopidogrel or prasugrel to inhibit platelet function irreversibly. Careful consideration should be given to the timing of administration of an oral P2Y(12) antagonist following cangrelor infusion.
Concomitant administration of cangrelor with the thienopyridine antiplatelet drugs clopidogrel or prasugrel decreases the antiplatelet effect of clopidogrel and prasugrel by blocking P2Y12-receptor binding of the active metabolites of these drugs. Oral maintenance antiplatelet therapy with clopidogrel or prasugrel should not be administered until the cangrelor infusion has been discontinued.

[1]. Intravenous cangrelor as a peri-procedural bridge with applied uses in ischemic events. Ann Transl Med. 2019;7(17):408.

[2]. Cangrelor alleviates bleomycin-induced pulmonary fibrosis by inhibiting platelet activation in mice. Mol Immunol. 2020;120:83-92.

[3]. Contribution of platelet P2Y12 receptors to chronic Complete Freund's adjuvant-induced inflammatory pain. J Thromb Haemost. 2017;15(6):1223-1235.

Cangrelor tetrasodium is an organic sodium salt that is the tetrasodium salt of cangrelor. Used as an intravenous antiplatelet drug that prevents formation of harmful blood clots in the coronary arteries. It has a role as a platelet aggregation inhibitor and a P2Y12 receptor antagonist. It contains a cangrelor(4-).
Cangrelor Tetrasodium is the tetrasodium salt form of cangrelor, an inhibitor of the platelet adenosine diphosphate (ADP) P2Y12 receptor (P2Y12R), with antiplatelet activity. Upon administration, cangrelor selectively and reversibly binds to P2Y12R, and blocks the platelet signaling pathway. This inhibits the activation of the glycoprotein complex GPIIb/IIIa, fibrinogen binding to platelets and platelet adhesion and aggregation.

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Drug Indication
Kengrexal, co-administered with acetylsalicylic acid (ASA), is indicated for the reduction of thrombotic cardiovascular events in adult patients with coronary artery disease undergoing percutaneous coronary intervention (PCI) who have not received an oral P2Y12 inhibitor prior to the PCI procedure and in whom oral therapy with P2Y12 inhibitors is not feasible or desirable.

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Due to the increasing number of patients on antiplatelet therapy for cardiovascular and neurological conditions, it can be challenging to manage these patients peri-operatively. It is critical to prevent ischemia and thrombosis, and at the same time decrease the risk of bleeding, necessitating the need for a successful bridging therapy. Cangrelor looks promising as a bridging therapy with its distinctive pharmacokinetic profile with fast activity and easy reversibility. However, large prospective studies are required to delineate clear guidelines to identify the patient population that would receive maximum benefit from bridging antiplatelet therapy, determine optimal dosing and titration, monitoring therapy, and manage adverse events. Although guidelines recommend IV bridge therapy in these settings, no agent currently has FDA-approval for this indication and positive, randomized controlled data for GPIs in this setting is also lacking. Cangrelor bridging therapy appears to have advantages over the previous standard using GPIs due to its faster offset and non-renal clearance. Future research is warranted for use of cangrelor in special populations, such as those with CAD on DAPT as a bridge to LVAD implantation. Overall, in this complex era of advancing medical technologies, therapies such as cangrelor may mitigate thrombotic and bleeding risks in the peri-operative period. [1]

C17H21CL2F3N5NA4O12P3S2

864.27

862.876

C, 23.63; H, 2.45; Cl, 8.20; F, 6.59; N, 8.10; Na, 10.64; O, 22.21; P, 10.75; S, 7.42

163706-36-3

Cangrelor; 163706-06-7

10260031

White to off-white solid powder

979ºC at 760 mmHg

545.9ºC

0mmHg at 25°C

4.676

347.26

3

21

14

48

1110

4

CSCCNC1=C2C(=NC(=N1)SCCC(F)(F)F)N(C=N2)C3C(C(C(O3)COP(=O)([O-])OP(=O)(C(P(=O)([O-])[O-])(Cl)Cl)[O-])O)O.[Na+].[Na+].[Na+].[Na+]

COWWROCHWNGJHQ-OPKBHZIBSA-J

InChI=1S/C17H25Cl2F3N5O12P3S2.4Na/c1-43-5-3-23-12-9-13(26-15(25-12)44-4-2-16(20,21)22)27(7-24-9)14-11(29)10(28)8(38-14)6-37-42(35,36)39-41(33,34)17(18,19)40(30,31)32;;;;/h7-8,10-11,14,28-29H,2-6H2,1H3,(H,33,34)(H,35,36)(H,23,25,26)(H2,30,31,32);;;;/q;4*+1/p-4/t8-,10-,11-,14-;;;;/m1..../s1

tetrasodium;[dichloro(phosphonato)methyl]-[[(2R,3S,4R,5R)-3,4-dihydroxy-5-[6-(2-methylsulfanylethylamino)-2-(3,3,3-trifluoropropylsulfanyl)purin-9-yl]oxolan-2-yl]methoxy-oxidophosphoryl]oxyphosphinate

ARC69931; AR-C69931; AR C69931; Cangrelor; AR-C69931MX; Cangrelor tetrasodium; 163706-36-3; AR-C69931MX; kengrexal; Cangrelor Tetrasodium [USAN]; Cangrelor tetrasodium salt; 5'-Adenylic acid, N-[2-(methylthio)ethyl]-2-[(3,3,3-trifluoropropyl)thio]-, anhydride with P,P'-(dichloromethylene)bis[phosphonic acid], sodium salt (1:1:4); 2144G00Y7W; AR C69931MX; ARC69931MX; Cangrelor tetrasodium salt

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中（例如氮气保护），避免吸湿/受潮和光照。

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

H2O : ~125 mg/mL (~144.63 mM)
DMSO : ~12.5 mg/mL (~14.46 mM)

配方 1 中的溶解度: 1.25 mg/mL (1.45 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。
例如，若需制备1 mL的工作液，可将100 μL 12.5 mg/mL澄清的DMSO储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: 1.25 mg/mL (1.45 mM) 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; 超声助溶.
例如，若需制备1 mL的工作液，可将 100 μL 12.5 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。

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配方 3 中的溶解度: 1.25 mg/mL (1.45 mM) 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; 超声助溶.
例如，若需制备1 mL的工作液，可将 100 μL 12.5 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 100 mg/mL (115.70 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶.

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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；
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