BPTU (BMS-646786) is a non-nucleotide P2Y₁ receptor allosteric antagonist with antithrombotic activity. BPTU is able to block the P2Y1 receptor located at the neuromuscular junction of the gastrointestinal tract^[1][2].

P2Y₁^[1]

BPTU blocks the supramaximal fast inhibitory junction potentials (fIJP) in a concentration-dependent manner both in the rat and mouse colon. The EC₅₀ of BPTU is approximately 0.3 μM and 0.06 μM for the rat and mouse colon, respectively. In the rat colon, addition of the P2Y agonist ADPβS at 10 μM significantly reduces spontaneous contractions to a 43.2±13.4% (N=5) (P=0.0002), and this reduction is blocked by 15 min incubation with BPTU at a concentration of 3 μM (93.3±5.1%). Similar results are obtained in the murine colon where ADPβS at 10 μM reduces the area under the curve (AUC) of contractions to a 15.8±5.1% (N=4) (P<0.0001) and its effect is reversed with BPTU at 3 μM (82.7±3.6%). Addition of MRS2365, a selective P2Y1 agonist, at a concentration of 5 μM significantly reduces spontaneous contractions to a 21.2±4.8% (N=5) (P=0.0002) in the murine colon, and this reduction is blocked by 15 min incubation with BPTU at a concentration of 3 μM (93.1±3.8%). The blockage of the MRS2365-induced response by BPTU at 3 μM also occurs in control conditions (N=5) (10.2±5.5% vs. 86.7±5.0%)^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Uptake of BPTU from the peritoneal cavity is relatively rapid. Blood boron levels are maximal within 1 h after administration. After only 1 h, a boron tumor-to-blood ratio above 1 is found for BPTU in pigmented tumors, which is indicative of drug retention. This is not seen in the non-pigmented tumor variant, in which tumor boron levels closely follow blood levels. Up to 24 h, Borocaptate sodium (BSH) exhibits no selective retention in either tumor, but achieves higher maximum tumor boron concentrations than BPTU as a result of the administration of higher amounts of boron. During the tissue distribution phase, liver-to-kidney boron concentration ratios range from 2 to 4 for BSH and from 0.5 to 1 for BPTU^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

445.43

C₂₃H₂₂F₃N₃O₃

870544-59-5

Room temperature in continental US; may vary elsewhere.

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : ≥ 33.3 mg/mL (74.76 mM)

* “≥” means soluble, but saturation unknown.

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.5 mg/mL (5.61 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (5.61 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (5.61 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (5.61 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Noemí Mañé, et al. BPTU, an allosteric antagonist of P2Y1 receptor, blocks nerve mediated inhibitory neuromuscular responses in the gastrointestinal tract of rodents. Neuropharmacology. 2016 Nov;110(Pt A):376-385.

  [2]. Dandan Zhang, et al. Two disparate ligand binding sites in the human P2Y1 receptor. Nature. 2015 Apr 16; 520(7547): 317–321.

Electrophysiological experiments are performed with colonic rat and mouse strips. Inhibitory junction potentials (IJP) are elicited by electrical field stimulation (EFS) using two silver chloride plates placed 1.5 cm apart perpendicular to the longitudinal axis of the preparation. The protocol consists of single pulse trains of EFS (0.4 ms pulse duration) at increasing voltages (8, 12, 16, 20, 24, 28, 32, 36, 40 V). The voltage responsible for the supramaximal response is used to elicit single pulses during incubation with BPTU at increasing concentrations (1×10^-8 M, 1×10^-7 M, 3×10^-7 M, 1×10^-6 M and 3×10^-6 M). Another train of single pulses at increasing voltages is elicited after the highest dose of BPTU. The amplitude of the IJP (mV) is measured considering it as the difference between the maximal hyperpolarization and the resting membrane potential (RMP)^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Mice are given BPTU intraperitoneally at doses of 3.15 mg of boron per kg body weight. Injection volumes range from 0.25 to 0.5 mL for both intravenous and intraperitoneal administrations. Six mice are not given any drug to allow measurement of boron background levels. Animals are killed with carbon dioxide 0.2, 0.4, 1, 2, 4, 24 and 48 h after drug administration and samples are taken from tumor, blood, skin, muscle, brain, kidneys and liver. Tumor tissue from mice bearing B16.013 tumor is checked by eye for the absence of pigmentation. BPTU is also given in a multiple dose scheme. Every 2 h 0.4 to 0.5 mL of the above-mentioned BPTU solution is given intraperitoneally (4×3.15 mg/kg boron). Twenty-four hours after the last administration, the animals are sacrificed and samples are taken^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Noemí Mañé, et al. BPTU, an allosteric antagonist of P2Y1 receptor, blocks nerve mediated inhibitory neuromuscular responses in the gastrointestinal tract of rodents. Neuropharmacology. 2016 Nov;110(Pt A):376-385.

  [2]. Dandan Zhang, et al. Two disparate ligand binding sites in the human P2Y1 receptor. Nature. 2015 Apr 16; 520(7547): 317–321.