PD 146176
(Synonyms: NSC168807) 目录号 : GC12205
PD 146176 是一种有效的选择性网织红细胞 15-LOX-1 抑制剂。
Cas No.:4079-26-9
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
murine melanoma cell line B16F10 |
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Preparation Method |
1 × 104 cells per well were seeded in each well of a 96-well tissue culture plate and left overnight in the incubator. The cells were incubated with PD 146176 for 24, 48, or 72 h similar to the MTT assay. |
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Reaction Conditions |
0.5, 1, 2 , 10, 20, 40µM for 24, 48, 72 hours |
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Applications |
PD 146176 induced cell cycle arrest in G1 phase at 8 h with decrease of cells in S and G2/M phase. The effect on cell morphology is visible observed after 16 h with cells becoming smaller and rounded. |
| Animal experiment [2]: | |
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Animal models |
C57BL/6 mice |
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Preparation Method |
One group was untreated and ate ground rodent chow for 7 days while the experimental group was fed ground rodent chow with the selective PD 146176 added at a concentration to achieve a dose of about 400 mg/kg/day |
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Dosage form |
Fed with diet , 400 mg/kg/day, 7 days |
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Applications |
The mice that were fed PD 146176 lost significantly more weight at 3-5 days after starting dextran sodium sulfate, compared to the corresponding day for the control mice. |
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References: [1]: Da-Costa-Rocha I, Prieto J M. In vitro effects of selective COX and LOX inhibitors and their combinations with antineoplastic drugs in the mouse melanoma cell line B16F10[J]. International journal of molecular sciences, 2021, 22(12): 6498. | |
PD 146176 is one of the potent and selective inhibitors of reticulocyte 15-LOX-1 [1]. PD 146176 inhibited the activity of h-15-LOX-1 with IC50 value of 16 ± 2.5μM,and Ki value of 3.9 ± 0.6μM [2].
PD 146176 (1 μg ml-1) inhibited the induction of both arginase-1 and mannose receptor mRNA in both rIL-4-treated and RSV-infected WT macrophages, whereas enhancing the induction of COX-2 mRNA [3]. PD 146176 suggested that it causes strong cell cycle arrest in G1 phase. PD 146176 at its IC50, did not show any inhibitory effect on cell directional migration but greatly increased the activity of the caspases in B16F10 cells [4].PD 146176 significantly prevented glutamate-induced cell death in a concentration-dependent manner. PD 146176 fully protected HT-22 cells against glutamate toxicity at a concentration of 0.5 μM and significantly reduced the annexin-V/propidium iodide-positive cells [5].
PD 146176 treated 3xTg mice had significant reductions in Aβ peptide levels, amyloid plaque burden, tau phosphorylation, and insoluble tau deposition in comparison with controls [6,7]. AD model mice in the control group showed a worsening of memory and learning abilities, whereas mice receiving PD 146176 were undistinguishable from wild-type mice [7].
References:
[1]. Orafaie A, Mousavian M, Orafai H, et al. An overview of lipoxygenase inhibitors with approach of in vivo studies[J]. Prostaglandins & Other Lipid Mediators, 2020, 148: 106411.
[2]. Eleftheriadis N, Thee S, Te Biesebeek J, et al. Identification of 6-benzyloxysalicylates as a novel class of inhibitors of 15-lipoxygenase-1[J]. European Journal of Medicinal Chemistry, 2015, 94: 265-275.
[3]. Shirey K A, Lai W, Pletneva L M, et al. Role of the lipoxygenase pathway in RSV-induced alternatively activated macrophages leading to resolution of lung pathology[J]. Mucosal immunology, 2014, 7(3): 549-557.
[4]. Da-Costa-Rocha I, Prieto J M. In vitro effects of selective COX and LOX inhibitors and their combinations with antineoplastic drugs in the mouse melanoma cell line B16F10[J]. International journal of molecular sciences, 2021, 22(12): 6498.
[5]. Tobaben S, Grohm J, Seiler A, et al. Bid-mediated mitochondrial damage is a key mechanism in glutamate-induced oxidative stress and AIF-dependent cell death in immortalized HT-22 hippocampal neurons[J]. Cell Death & Differentiation, 2011, 18(2): 282-292.
[6]. Oddo S, Caccamo A, Shepherd J D, et al. Triple-transgenic model of Alzheimer's disease with plaques and tangles: intracellular Aβ and synaptic dysfunction[J]. Neuron, 2003, 39(3): 409-421.
[7]. Di Meco A, Li J G, Blass B E, et al. 12/15-Lipoxygenase inhibition reverses cognitive impairment, brain amyloidosis, and tau pathology by stimulating autophagy in aged triple transgenic mice[J]. Biological psychiatry, 2017, 81(2): 92-100.
PD 146176 是一种有效的选择性网织红细胞 15-LOX-1 抑制剂[1]。 PD 146176 抑制 h-15-LOX-1 的活性，IC50 值为 16 ± 2.5μM，Ki 值为 3.9 ± 0.6μM [2]。
PD 146176（1 μg ml-1）抑制 rIL-4 处理和 RSV 感染的 WT 巨噬细胞中精氨酸酶-1 和甘露糖受体 mRNA 的诱导，同时增强 COX-2 mRNA 的诱导 [3]. PD 146176 表明它会在 G1 期引起强烈的细胞周期停滞。 IC50 的 PD 146176 对细胞定向迁移没有任何抑制作用，但大大增加了 B16F10 细胞中半胱天冬酶的活性[4]。PD 146176 在一定浓度下显着阻止谷氨酸诱导的细胞死亡-依赖的方式。 PD 146176 在 0.5 μM 浓度下完全保护 HT-22 细胞免受谷氨酸毒性，并显着减少膜联蛋白-V/碘化丙锭阳性细胞[5]。
与对照组相比，PD 146176 处理的 3xTg 小鼠的 Aβ 肽水平、淀粉样斑块负荷、tau 磷酸化和不溶性 tau 沉积显着降低[6,7]。对照组的 AD 模型小鼠表现出记忆和学习能力的恶化，而接受 PD 146176 的小鼠与野生型小鼠没有区别[7]。
| Cas No. | 4079-26-9 | SDF | |
| 别名 | NSC168807 | ||
| 化学名 | 6,11-dihydrothiochromeno[4,3-b]indole | ||
| Canonical SMILES | C12=C(C3=CC=CC=C3SC2)NC4=CC=CC=C14 | ||
| 分子式 | C15H11NS | 分子量 | 237.32 |
| 溶解度 | DMF: 10 mg/ml,DMF:PBS (pH 7.2) (1:5): 0.15 mg/ml,DMSO: 10 mg/ml,Ethanol: 2 mg/ml | 储存条件 | Store at -20°C |
| General tips | 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 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 4.2137 mL | 21.0686 mL | 42.1372 mL |
| 5 mM | 0.8427 mL | 4.2137 mL | 8.4274 mL |
| 10 mM | 0.4214 mL | 2.1069 mL | 4.2137 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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Glutathione peroxidase 4 senses and translates oxidative stress into 12/15-lipoxygenase dependent- and AIF-mediated cell death
Oxidative stress in conjunction with glutathione depletion has been linked with various acute and chronic degenerative disorders, yet the molecular mechanisms have remained unclear. In contrast to the belief that oxygen radicals are detrimental to cells and tissues by unspecific oxidation of essential biomolecules, we now demonstrate that oxidative stress is sensed and transduced by glutathione peroxidase 4 (GPx4) into a-yet-unrecognized cell-death pathway. Inducible GPx4 inactivation in mice and cells revealed 12/15-lipoxygenase-derived lipid peroxidation as specific downstream event, triggering apoptosis-inducing factor (AIF)-mediated cell death. Cell death could be entirely prevented either by alpha-tocopherol (alpha-Toc), 12/15-lipoxygenase inhibitors, or siRNA-mediated AIF silencing. Accordingly, 12/15-lipoxygenase-deficient cells were highly resistant to glutathione depletion. Neuron-specific GPx4 depletion caused neurodegeneration in vivo and ex vivo, highlighting the importance of this pathway in neuronal cells. Since oxidative stress is common in the etiology of many human disorders, the identified pathway reveals promising targets for future therapies.
In Vitro Effects of Selective COX and LOX Inhibitors and Their Combinations with Antineoplastic Drugs in the Mouse Melanoma Cell Line B16F10
The constitutive expression or overactivation of cyclooxygenase (COX) and lipoxygenase (LOX) enzymes results in aberrant metabolism of arachidonic acid and poor prognosis in melanoma. Our aim is to compare the in vitro effects of selective COX-1 (acetylsalicylic acid), COX-2 (meloxicam), 5-LOX (MK-886 and AA-861), 12-LOX (baicalein) and 15-LOX (PD-146176) inhibition in terms of proliferation (SRB assay), mitochondrial viability (MTT assay), caspase 3-7 activity (chemiluminescent assay), 2D antimigratory (scratch assay) and synthesis of eicosanoids (EIA) in the B16F10 cell line (single treatments). We also explore their combinatorial pharmacological space with dacarbazine and temozolomide (median effect method). Overall, our results with single treatments show a superior cytotoxic efficacy of selective LOX inhibitors over selective COX inhibitors against B16F10 cells. PD-146176 caused the strongest antiproliferation effect which was accompanied by cell cycle arrest in G1 phase and an >50-fold increase in caspases 3/7 activity. When the selected inhibitors are combined with the antineoplastic drugs, only meloxicam provides clear synergy, with LOX inhibitors mostly antagonizing. These apparent contradictions between single and combination treatments, together with some paradoxical effects observed in the biosynthesis of eicosanoids after FLAP inhibition in short term incubations, warrant further mechanistical in vitro and in vivo scrutiny.
Attenuation of diet-induced atherosclerosis in rabbits with a highly selective 15-lipoxygenase inhibitor lacking significant antioxidant properties
1. 15-Lipoxygenase (15-LO) has been implicated in the pathogenesis of atherosclerosis because of its localization in lesions and the many biological activities exhibited by its products. To provide further evidence for a role of 15-LO, the effects of PD 146176 on the development of atherosclerosis in cholesterol-fed rabbits were assessed. This novel drug is a specific inhibitor of the enzyme in vitro and lacks significant non specific antioxidant properties. 2. PD 146176 inhibited rabbit reticulocyte 15-LO through a mixed noncompetitive mode with a Ki of 197 nM. The drug had minimal effects on either copper or 2,2'-azobis(2-amidinopropane)hydrochloride (ABAP) induced oxidation of LDL except at concentrations 2 orders higher than the Ki. 3. Control New Zealand rabbits were fed a high-fat diet containing 0.25% wt./wt. cholesterol; treated animals received inhibitor in this diet (175 mg kg-1, b.i.d.). Plasma concentrations of inhibitor were similar to the estimated Ki (197 nM). During the 12 week study, there were no significant differences in weight gain haematocrit, plasma total cholesterol concentrations, or distribution of lipoprotein cholesterol. 4. The drug plasma concentrations achieved in vivo did not inhibit low-density lipoprotein (LDL) oxidation in vitro. Furthermore, LDL isolated from PD 146176-treated animals was as susceptible as that from controls to oxidation ex vivo by either copper or ABAP. 5. PD 146176 was very effective in suppressing atherogenesis, especially in the aortic arch where lesion coverage diminished from 15 +/- 4 to 0% (P < 0.02); esterified cholesterol content was reduced from 2.1 +/- 0.7 to 0 micrograms mg-1 (P < 0.02) in this region. Immunostainable lipid-laden macrophages present in aortic intima of control animals were totally absent in the drug-treated group. 6. Results of these studies are consistent with a role for 15-LO in atherogenesis.
Lipoxin biosynthesis in inflammatory bowel disease
Background and aims: Lipoxins are anti-inflammatory lipid mediators that are produced in gut mucosa, which serve to limit and resolve persistent inflammation. The purpose of this study was to evaluate colonic lipoxin biosynthesis in patients with ulcerative colitis (UC) to establish a possible biochemical basis for persistent inflammation in UC.
Methods: Colonic mucosa from patients with UC or organ donors (controls) was placed into tissue culture for 90 min. The conditioned media was assayed (ELISA) for lipoxin A4 (LXA) and the biologically active isomer 15-epi-LXA4 (aspirin triggered lipoxin, ATL). Mucosal tissue 15-lipoxygenase protein was determined by Western blot.
Results: Patient colonic mucosa produced significantly lower (12-fold) amounts of LXA, relative to organ donors. This occurred irregardless of patient steroid treatment. However, patient tissue responded to in vitro aspirin by synthesizing biologically active ATL. For the first time, human colonic mucosa was found to synthesize 15-lipoxygenase-2, an epithelial-derived isoenzyme used for lipoxin synthesis. These levels were significantly lower in UC patients compared to the control tissue. Finally, mice chronically treated with a putative selective 15-lipoxygenase inhibitor (PD 146176) experienced significantly worse intestinal function during experimental colitis, relative to untreated mice.
Conclusion: Colonic mucosa from UC patients demonstrated defective lipoxin biosynthesis, which may contribute to the inability of these patients to resolve persistent colonic inflammation.
Arachidonic acid epoxygenase and 12(S)-lipoxygenase: evidence of their concerted involvement in ductus arteriosus constriction to oxygen
Oxygen promotes closure of the ductus arteriosus at birth. We have previously presented a scheme for oxygen action with a cytochrome P450 (CYP450) hemoprotein and endothelin-1 (ET-1) being, respectively, sensor and effector, and a hypothetical monooxygenase product serving as a coupling link. We have also found in the vessel arachidonic acid (AA) 12(S)-lipoxygenase (12-lipoxygenase) undergoing upregulation at birth. Here, we examined the feasibility of a sensor-to-effector messenger originating from AA monooxygenase and 12-lipoxygenase pathways. The epoxygenase inhibitor, N-methylsulfonyl-6-(2-)hexanamide, suppressed the tonic contraction of ductus to oxygen. A similar effect was obtained with 12-lipoxygenase inhibitors baicalein and PD 146176. By contrast, none of the inhibitors modified the endothelin-1 contraction. Furthermore, an AA ω-hydroxylation product, 20-hydroxyeicosatetraenoic acid (20-HETE), reportedly responsible for oxygen contraction in the systemic microvasculature, had no such effect on the ductus. We conclude that AA epoxygenase and 12-lipoxygenase jointly produce a hitherto uncharacterized compound acting as oxygen messenger in the ductus.