Dicoumarol (Dicumarol)   中文名称: 双香豆素

NQO1 (IC50 = 0.37 μM); PDK1 (IC50 = 19.42 μM)

双香豆素用作 PDK1 和 NAD (P) H:醌氧化还原酶 1 (NQO1) 的对照，IC50 值分别为 19.42±0.032 μM 和 0.37±0.15。双香豆素旨在阻止 PDK1 的作用。暴露于 200 μM 双香豆素后，PDK1 的酶活性几乎下降了 94%。双香豆素在 100 μM 时使 p-PDHA1 水平降低 26%，在 200 μM 时降低 72%，而 PDHA1 的总体水平没有显着变化。浓度为 100 和 200 μM 的双香豆素均具有强烈诱导作用。同样，根据膜联蛋白 V+PI+ 细胞的流式细胞术检查，用 100 μM 和 200 μM 双香豆素处理分别产生约 20.87% 和 24.94%。移植细胞，尤其是经过多次溶剂处理后[2]。此外，值得注意的是，当用众所周知的 NQO1 双香豆素处理时，MCF-7-TAMR 细胞的他莫昔芬反应表型发生逆转 [3]。

当给予100mg/kg二氯乙酸(DCA)、30mg/kg双香豆素和50mg/kg双香豆素时，与来自溶剂组或沙漠组的肿瘤相比，肿瘤重量和体积显着减少。当用双香豆素处理的 SKOV3 异种移植物与载体或载体组中的肿瘤进行比较时，总 caspase-3 和总抗聚（ADP-核糖）聚合酶（PARP）显着下降 [2]。

使用标准 MTT 测定检查体外细胞活力。在 96 孔板中，每孔接种 8000 个 SKOV3 或 A2780 细胞。第二天，每个孔中填充浓度逐渐升高的双香豆素 (DIC)，并将板孵育 24 小时。然后将板再孵育 4 小时，然后在每个孔中加入 10 μL 10 mg/mL MTT 试剂的磷酸盐缓冲盐水 (PBS) 溶液。摇动板 5 分钟后，将甲臜晶体溶解在 150 μL DMSO 中后，读数器测量 570 nm 处的光密度[2]。

We use twenty-five female BALB/c-nu mice that are 15 g in weight and 5 to 6 weeks old. The upper flank is subcutaneously injected with a total of 1 107 SKOV3 cells. The nude mice are randomized into five groups (n=5/group) after 10 days, when the tumor volume reaches roughly 100 mm3, and are treated intraperitoneally (i.p.) every other day for a total of 12 days with the following medications: Dichloroacetate (DCA) group received 100 mg/kg of DCA; Dicoumarol (DIC)-30 group received 30 mg/kg of Dicoumarol; and Dicoumarol-50 group received 50 mg/kg of Dicoumarol. Control groups received 0.2 mL of 0.9% NaCl, 1 mM NaOH, and Dichloroacetate (DCA) group received 100 mg/kg of DCA. Every other day until sacrifice (day 12 following the initial treatment), the body weights and tumor volumes of each mouse are measured[2].

Absorption, Distribution and Excretion
Considerable individual variation in t/2 of dicumarol has been attributed to genetic factors. ... dicumarol...hydroxylated to inactive compounds by enzymes of hepatic endoplasmic reticulum. These metabolites and traces of parent drugs are excreted in urine. Some unabsorbed dicumarol appears in feces.
In man, absorption of dicumarol from gi tract is slow and erratic. ... There is considerable variation in absorption from one individual to another. Within circulation...almost entirely but loosely bound to plasma albumin, & only small percentage of total plasma concentration is represented by unbound drug. ... Appreciable amount ...found in erythrocytes, but little or none is present in cerebrospinal fluid. ...accumulate/s/ mainly in lung, liver, spleen and kidney.
Whole-body autoradiography of rats given anticoagulant, [(14)C]-dicumarol by intracardiac injection, indicated that (14)C distributed in most tissues, maximally in liver, lungs, heart, and kidneys. After 24 hr, (14)C levels were high in intestinal tract owing, presumably, to biliary excretion. Initially, iv dose of dicoumarol was more readily excreted in bile than in urine; in 3 hr, 4% was eliminated in bile and less than 0.4% in urine.
...71% of iv dose of...[(14)C]-dicoumarol, was excreted in feces and 23% in urine in 5 days.

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Metabolism / Metabolites
Dicoumarol is not conjugated in either man or dog...
Dicumarol...hydroxylated to inactive cmpd by enzymes of hepatic endoplasmic reticulum.
Despite their structural similarity to coumarin, the anticoagulants dicumarol and warfarin do not appear to be substrates for CYP2A6. The overall rate of dicumarol metabolism varied approx 5 fold among the human liver microsomal samples, but this variation correlated poorly (r2= 0.126) with the variation observed in CYP2A6 levels and hydroxycoumarin levels.

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Biological Half-Life
1-2 days
...T/2 of dicumarol is dose dependent, ranging from 10 hr at low dosage to 30 hr at high dosage.
Dicumarol has a dose dependent plasma half-life (one to two days); therapy is therefore somewhat difficult to control and frequent monitoring is usually indicated.
Elimination half-life: 1 to 2 days

Toxicity Summary
Dicoumarol is an anticoagulant that competitively inhibits vitamin K, preventing the formation of prothrombin. It does this by inhibiting the enzyme NAD(P)H:quinone oxidoreductase-1, which is required for the reduction of vitamin K to its hydroquinone. Reduced vitamin K is a cofactor needed in the conversion of prothrombin precursor protein to active prothrombin, an essential protein for blood clotting. In addition, inhibition of NAD(P)H:quinone oxidoreductase-1 induces the generation of superoxide anion radicals that inhibit cell growth. Dicoumarol can also potently and reversible inhibit gap junctional intercellular communication, though the precise mechanism is unknown. (L1960, A2994, A2995, A2996, A2997)

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Toxicity Data
LD₅₀=233 mg/kg (orally in mice)
LD₅₀=250 mg/kg (orally in rats)

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Interactions
Dicumarol admin prolongs the half-life of chlorpropamide and phenytoin, resulting in hypoglycemia in the case of chlorpropamide and an increased plasma drug concn in the case of phenytoin.
...Sulfonamides (esp long-acting ones) displace dicumarol from plasma proteins & hence incr effect.
...Carbon tetrachloride & chloral hydrates are strong potentiators of its anticoagulant effects.
Antipyrine (in vivo) & sulfinpyrazone (in vitro) have been reported to interact with warfarin and should be used cautiously in pt receiving anticoagulants. /anticoagulants/
For more Interactions (Complete) data for DICUMAROL (20 total), please visit the HSDB record page.

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Non-Human Toxicity Values
LD50 Mouse iv 42 mg/kg
LD50 Mouse sc 50 mg/kg
LD50 Mouse ip 91 mg/kg
LD50 Mouse oral 233 mg/kg
For more Non-Human Toxicity Values (Complete) data for DICUMAROL (6 total), please visit the HSDB record page.

[1]. Affinity-based small fluorescent probe for NAD(P)H:quinone oxidoreductase 1 (NQO1). Design, synthesis and pharmacological evaluation. Eur J Med Chem. 2017 Feb 15;127:828-839.

[2]. Dicumarol inhibits PDK1 and targets multiple malignant behaviors of ovarian cancer cells. PLoS One. 2017 Jun 15;12(6):e0179672.

[3]. Mitochondrial "power" drives tamoxifen resistance: NQO1 and GCLC are new therapeutic targets in breast cancer. Oncotarget. 2017 Mar 2;8(12):20309-20327.

Therapeutic Uses
Anticoagulants; Enzyme Inhibitors; Uncoupling Agents
Anticoagulants are indicated for prophylaxis and/or treatment of venous (or arterial) thrombosis (and its extension) and pulmonary embolism /Not included in US product labeling/, deep vein thrombosis (DVT) or pulmonary embolism (treatment). Oral anticoagulants are used during and following initial heparin therapy to decrease the risk of extension, recurrence, or death. /Anticoagulants; Included in US product labeling/
Oral anticoagulants are used to prevent thromboembolic complications after surgery, although low-dose subcutaneous heparin is used more commonly. /Anticoagulants; Included in US product labeling/
Anticoagulants are indicated for prophylaxis and/or treatment of thromboembolic complications (ischemic stroke) associated with atrial fibrillation. They are strongly recommended in patients at high risk of stroke (including patients with recent stroke, transient ischemic attack, or systemic embolism; poor left ventricular function; age over 75 years; hypertension; rheumatic mitral valve disease; mechanical or tissue prosthetic heart valves.) /Anticoagulants; Included in US product labeling/
For more Therapeutic Uses (Complete) data for DICUMAROL (9 total), please visit the HSDB record page.

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Drug Warnings
Contraindications to oral anticoagulants include pre-existing or coexisting abnormalities of blood coagulation, active bleeding, recent or imminent surgery of the central nervous system or eye, diagnostic or therapeutic procedures with potential for uncontrollable bleeding including lumbar puncture, malignant hypertension, peptic ulceration, pregnancy, threatened abortion, intrauterine device, cerebrovascular hemorrhage, and bacterial endocarditis. Relative contraindications include thrombocytopenia, pericarditis, pericardial effusions, and unreliability of the patient or of patient supervision. /Oral anticoagulants/
Most commonly, oral anticoagulant-induced bleeding is minor and consists of bruising, hematuria, epistaxis, conjunctival hemorrhage, minor gastrointestinal bleeding, bleeding from wounds and sites of trauma, and vaginal bleeding. More serious major or fatal bleeding is most commonly gastrointestinal, intracranial, vaginal, retroperitoneal, or related to a wound or site of trauma, although a large variety of other sites of bleeding have been reported. Intracranial bleeding occurs most frequently in patients receiving oral anticoagulants for cerebrovascular disease and most commonly presents as a subdural hematoma, often unassociated with head trauma. Fatal gastrointestinal bleeding is most commonly from a peptic ulcer, although any gastrointestinal lesion may be a potential source of major bleeding. Overall, a bleeding lesion can be identified in about two thirds of cases of oral anticoagulants-related hemorrhage. /Oral anticoagulants/
Overall, the bleeding rate of oral anticoagulant therapy is influenced by several factors: the intensity of anticoagulation, either intentionally or inadvertent; the underlying clinical disorder for which anticoagulant therapy is used (with bleeding occurring most frequently in ischemic cerebrovascular disease and venous thromboembolism; and, with bleeding occurring most commonly in the elderly; the presence of adverse drug interactions or comorbid factors such as clinical states potentiating warfarin action, pre-existing hemorrhagic diathesis, malignancy, recent surgery, trauma, or pre-existing potential bleeding sites (e.g., surgical wound, peptic ulcer, recent cerebral hemorrhage, carcinoma of colon); the simultaneous use of aspirin (but not of dipyridamole); and patient reliability (e.g., increased bleeding in alcoholics not due to ethanol-warfarin drug interaction but rather to unreliability of drug intake). /Oral anticoagulants/
Spontaneous abortion and stillbirth have occurred, as well as low birth weight and growth retardation. In addition, fetal or neonatal hemorrhage, fetal death from hemorrhage in utero, and increased risk of maternal hemorrhage during the second and third trimesters have been reported. There is some evidence that embryopathy occurs only with oral anticoagulant administration between the 6th and 12th weeks of gestation. /Anticoagulants/
For more Drug Warnings (Complete) data for DICUMAROL (34 total), please visit the HSDB record page.

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Pharmacodynamics
Dicumarol is an coumarin-like compound found in sweet clover. It is used as an oral anticoagulant and acts by inhibiting the hepatic synthesis of vitamin K-dependent coagulation factors (prothrombin and factors VII, IX, and X).

C19H12O6

336.29

336.063

C, 67.86; H, 3.60; O, 28.55

66-76-2

54676038

White to off-white solid powder

1.6±0.1 g/cm3

620.7±55.0 °C at 760 mmHg

290-292 °C(lit.)

231.9±25.0 °C

0.0±1.9 mmHg at 25°C

1.731

3.55

100.88

2

6

2

25

605

0

O1C(C(=C(C2=C([H])C([H])=C([H])C([H])=C12)O[H])C([H])([H])C1C(=O)OC2=C([H])C([H])=C([H])C([H])=C2C=1O[H])=O

DOBMPNYZJYQDGZ-UHFFFAOYSA-N

InChI=1S/C19H12O6/c20-16-10-5-1-3-7-14(10)24-18(22)12(16)9-13-17(21)11-6-2-4-8-15(11)25-19(13)23/h1-8,20-21H,9H2

4-hydroxy-3-[(4-hydroxy-2-oxochromen-3-yl)methyl]chromen-2-one

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

配方 1 中的溶解度: ≥ 1.67 mg/mL (4.97 mM) (饱和度未知) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% 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 16.7 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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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。