IBMX
(Synonyms: 3-异丁基-1-甲基黄嘌呤,3-Isobutyl-1-methylxanthine; Isobutylmethylxanthine) 目录号 : GC11730
IBMX 是一种非特异性磷酸二酯酶 (PDE) 抑制剂，可抑制 PDE3、PDE4 和 PDE5，IC50 值为 6.5、26.3 和 31.7 µm，但 PDE8A、PDE8B 和 PDE9 除外 。
Cas No.:28822-58-4
Sample solution is provided at 25 µL, 10mM.
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- Purity: >99.00%
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| Kinase experiment [1]: | |
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Preparation Method |
Purified protein containing PDE3, 4 or 5 enzyme was resuspended in 50 mM Tris HCl containing 5 mM MgCl2 (pH 7.5). Subsequently, the enzyme (11.5 mg ml 1, 10 μl) was incubated with Tris HCl (80 μl) and 10 μM cyclic GMP or cyclic AMP substrate (final concentration 1 μM containing 0.1 μCi [3H]-cyclic GMP or [3H]-cyclic AMP) was added. After 20 min at 37 °C, the samples were heated to 100 °C for 2 min. Ophiophagus hannah snake venom (10 mg ml 1, 10 μl) was then added and incubated at 37 °C for 10 min to convert the 5 -GMP and 5 -AMP to the uncharged nucleosides, guanosine and adenosine, respectively. An ion-exchange resin (200 μl) was added to bind all unconverted cyclic GMP or cyclic AMP. IBMX as inhibitor, used to inhibit PDE detection IC50. |
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Reaction Conditions |
IBMX 10 mM in DMSO |
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Applications |
IBMX is a broad-spectrum phosphodiesterase (PDE) inhibitor that inhibits PDE3,PDE4 and PDE5 with IC50 values of 6.5,26.3 and 31.7 μm, respectively. |
| Cell experiment [2]: | |
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Cell lines |
Cortical collecting duct (CCD) |
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Preparation Method |
Cells were grown in 24-well plates with 105 cells per well at confluence, monolayers were washed with phosphate buffer solution (PBS) and incubated with KMUP-1 (0.1-100μM) in the presence of 100uM IBMX for 20 min by adding 10% trichloroacetic acid (TCA) to stop the incubation. |
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Reaction Conditions |
100μM IBMX for 20 minutes |
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Applications |
IBMX(100 μM) activated ROMK channels and prevented further channel activation by ANG II or cGMP. |
| Animal experiment [3]: | |
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Animal models |
Six groups of male Sprague–Dawley rats were used (150‿80 g) |
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Preparation Method |
After 8 weeks of exposure to cold, 3 groups in each temperature condition received continuous intravenous infusion of 8-isobutyl-methylxanthine (8-IBMX) (PDE-1 inhibitor, 8.5 mg/kg per day),apocynin and vehicle. |
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Dosage form |
8.5 mg/kg/day for 1 week |
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Applications |
Treatments with IBMX and Apocynin significantly decrease cold-induced elevation of right ventricular (RV) systolic pressure although they do not decrease RV pressure to the warm control levels. IBMX or Apocynin significantly reduces medial layer thickness and increases lumen diameter of small PAs in cold-exposed rats. |
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References: [1]. Wu BN, Lin RJ,et,al. KMUP-1, a xanthine derivative, induces relaxation of guinea-pig isolated trachea: the role of the epithelium, cyclic nucleotides and K+ channels. Br J Pharmacol. 2004 Aug;142(7):1105-14. doi: 10.1038/sj.bjp.0705791. Epub 2004 Jul 5. PMID: 15237094; PMCID: PMC1575170. [2]. Wei Y, Liao Y,et,al. Angiotensin II type 2 receptor regulates ROMK-like K? channel activity in the renal cortical collecting duct during high dietary K? adaptation. Am J Physiol Renal Physiol. 2014 Oct 1;307(7):F833-43. doi: 10.1152/ajprenal.00141.2014. Epub 2014 Aug 6. PMID: 25100281; PMCID: PMC4187043. [3]. Crosswhite P, Sun Z. Inhibition of phosphodiesterase-1 attenuates cold-induced pulmonary hypertension. Hypertension. 2013 Mar;61(3):585-92. doi: 10.1161/HYPERTENSIONAHA.111.00676. Epub 2013 Jan 14. PMID: 23319544; PMCID: PMC4050371. |
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IBMX is a non-specific inhibitor of phosphodiesterase (PDE) inhibitor that inhibits PDE3, PDE4 and PDE5 with IC50 values of 6.5, 26.3 and 31.7 µm, except PDE8A, PDE8B and PDE9[1,7].
In cardiac H9c2 cells,IBMX reduced loss of δψm caused by H(2)O(2), indicating that inhibition of PDEs can prevent the mPTP opening. However, IBMX could not inhibit the pore opening in cells transfected with the constitutively active GSK-3β (GSK-3β-S9A) mutant, suggesting a critical role of GSK-3β in the action of IBMX. IBMX also reduced reperfusion injury in a GSK-3β dependent manner[5]. Increasing cAMP-signaling with Forskolin or IBMX significantly facilitated neuronal functional maturation. A continuous application of IBMX to the differentiation medium substantially increased the functional expression of voltage-gated Na(+) and K(+) channels, as well as neuronal firing frequency[6].
Chronic exposure to cold caused pulmonary arterial hypertension and increased phosphodiesterase-1C (PDE-1C) expression in pulmonary arteries (PAs) in rats. After 8-week exposure to cold, Treatment with 8-IBMX significantly attenuated the cold-induced increase in right ventricular pressure. Cold exposure also caused right-ventricular hypertrophy, whereas 8-IBMX reversed cold-induced right ventricular hypertrophy. 8-IBMX abolished cold-induced upregulation of PDE-1C in PAs[4]. In hyperglycemic rat, all test compounds decreased blood glucose and the effect of milrinone was potentiated by glybenclamide. Milrinone or IBMX did not change plasma insulin levels, but it was augmented by combination of milrinone and glybenclamide. In both species, liver glycogen storage was decreased by IBMX, mc5, mc6 or MCPIP, increased by mc2 and was not changed in the presence of mc1[3]. ANG II increased ROMK channel activity in CCDs isolated from high-K (HK)-fed but not normal K (NK)-fed rats. Pretreatment of CCDs with IBMX, a broad-spectrum PDE inhibitor, or cilostamide, a PDE3 inhibitor, abolished the stimulatory effect of ANG II on ROMK channels[2].
References:
[1]: Wu BN, Lin RJ, et,al. KMUP-1, a xanthine derivative, induces relaxation of guinea-pig isolated trachea: the role of the epithelium, cyclic nucleotides and K+ channels. Br J Pharmacol. 2004 Aug;142(7):1105-14. doi: 10.1038/sj.bjp.0705791. Epub 2004 Jul 5. PMID: 15237094; PMCID: PMC1575170.
[2]: Wei Y, Liao Y,et,al. Angiotensin II type 2 receptor regulates ROMK-like K? channel activity in the renal cortical collecting duct during high dietary K? adaptation. Am J Physiol Renal Physiol. 2014 Oct 1;307(7):F833-43. doi: 10.1152/ajprenal.00141.2014. Epub 2014 Aug 6. PMID: 25100281; PMCID: PMC4187043.
[3]: Hosseini A, Shafiee-Nick R, et,al. Differential metabolic effects of novel cilostamide analogs, methyl carbostiryl derivatives, on mouse and hyperglycemic rat. Iran J Basic Med Sci. 2012 Jul;15(4):916-25. PMID: 23493150; PMCID: PMC3586914.
[4]: Crosswhite P, Sun Z. Inhibition of phosphodiesterase-1 attenuates cold-induced pulmonary hypertension. Hypertension. 2013 Mar;61(3):585-92. doi: 10.1161/HYPERTENSIONAHA.111.00676. Epub 2013 Jan 14. PMID: 23319544; PMCID: PMC4050371.
[5]: Chanoit G, Zhou J,et,al. Inhibition of phosphodiesterases leads to prevention of the mitochondrial permeability transition pore opening and reperfusion injury in cardiac H9c2 cells. Cardiovasc Drugs Ther. 2011 Aug;25(4):299-306. doi: 10.1007/s10557-011-6310-z. PMID: 21643720.
[6]: Lepski G, Jannes CE, et,al. cAMP promotes the differentiation of neural progenitor cells in vitro via modulation of voltage-gated calcium channels. Front Cell Neurosci. 2013 Sep 19;7:155. doi: 10.3389/fncel.2013.00155. PMID: 24065885; PMCID: PMC3777016.
[7]: Soderling SH, Bayuga SJ, et,al. Identification and characterization of a novel family of cyclic nucleotide phosphodiesterases. J Biol Chem. 1998 Jun 19;273(25):15553-8. doi: 10.1074/jbc.273.25.15553. PMID: 9624145.
IBMX 是一种非特异性磷酸二酯酶 (PDE) 抑制剂，可抑制 PDE3、PDE4 和 PDE5，IC50 值为 6.5、26.3 和 31.7 µm，但 PDE8A、PDE8B 和 PDE9 除外 [1,7]。
在心脏 H9c2 细胞中，IBMX 减少了由 H(2)O(2) 引起的 δψm 损失，表明抑制 PDE 可以阻止 mPTP 开放。然而，IBMX 不能抑制转染组成型活性 GSK-3β 的细胞的孔隙开放； (GSK-3β-S9A) 突变体，表明 GSK-3β 的关键作用；在 IBMX 的行动中。 IBMX 还减少了 GSK-3β 中的再灌注损伤；依赖方式[5]。使用 Forskolin 或 IBMX 增加 cAMP 信号显着促进神经元功能成熟。将 IBMX 持续应用于分化培养基可显着提高电压门控 Na(+) 和 K(+) 通道的功能表达，以及神经元放电频率[6]。
长期暴露于寒冷导致大鼠肺动脉高压和磷酸二酯酶-1C (PDE-1C) 表达增加。暴露于寒冷 8 周后，用 8-IBMX 治疗显着减弱了寒冷引起的右心室压力增加。冷暴露也会导致右心室肥大，而 8-IBMX 可逆转冷诱导的右心室肥大。 8-IBMX 消除了 PA 中冷诱导的 PDE-1C 上调 [4]。在高血糖大鼠中，所有测试化合物均降低血糖，米力农的作用被格列本脲增强。米力农或 IBMX 不会改变血浆胰岛素水平，但米力农和格列本脲的组合会增加血浆胰岛素水平。在这两个物种中，IBMX、mc5、mc6 或 MCPIP 降低了肝糖原储存，mc2 增加了肝糖原储存，并且在 mc1 [3] 存在的情况下没有改变。 ANG II 增加了 CCD 中的 ROMK 通道活动，这些 CCD 是从高 K (HK) 喂养的大鼠中分离出来的，但不是正常的 K (NK) 喂养的大鼠。用广谱 PDE 抑制剂 IBMX 或 PDE3 抑制剂西洛酰胺预处理 CCD，消除了 ANG II 对 ROMK 通道的刺激作用[2]。
| Cas No. | 28822-58-4 | SDF | |
| 别名 | 3-异丁基-1-甲基黄嘌呤,3-Isobutyl-1-methylxanthine; Isobutylmethylxanthine | ||
| 化学名 | 3-isobutyl-1-methyl-1H-purine-2,6(3H,7H)-dione | ||
| Canonical SMILES | O=C(N(C)C1=O)N(C2=C1NC=N2)CC(C)C | ||
| 分子式 | C10H14N4O2 | 分子量 | 222.24 |
| 溶解度 | ≥ 9.45mg/mL in DMSO; Insoluble in Water | 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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1 mg | 5 mg | 10 mg |
| 1 mM | 4.4996 mL | 22.4982 mL | 44.9964 mL |
| 5 mM | 0.8999 mL | 4.4996 mL | 8.9993 mL |
| 10 mM | 0.45 mL | 2.2498 mL | 4.4996 mL |
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动物数量 | 只 |
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Cyclic adenosine monophosphate (cAMP) analog and phosphodiesterase inhibitor (IBMX) ameliorate human sperm capacitation and motility
Objectives: Human sperm quality is decreasing progressively. One of the foremost reasons for infertility is the failure in sperm capacitation. We examined the influence of a cAMP (cyclic-adenosine mono phosphate analog)+IBMX (3-isobutyl-1-methylxanthine) on the motility and capacitation rate of human sperm over time. Material and methods: Samples were gotten from 20 asthenozoospermic infertile patients referring to the Academic Center for Education, Culture and Research unit of the infertility research center, Qom, Iran. Samples were processed with a Density Gradient Centrifuging. Spermatozoa were divided into 4 groups: control, experimental 1, 2 and 3 (E1, E2, E3) based on the dose/time schedules (cAMP 5mmol+IBMX 0.2mmol/2, 4, and 6h, respectively). The computer-assisted sperm analysis and chlortetracycline assays were used to measure sperm motility and capacitation. Results: After incubation with a cAMP analog and IBMX, the levels of progressive motile sperms considerably improved in all experimental groups compared to the control group (E1=18.89±7.1, E2=30±9.7, E3=26.3±9.6 vs Control=10.28±6.2, P<0.05) especially in E2 group (P<0.05), indicating a greater effect of db cAMP (5mmol) and IBMX (0.2mmol) for 4h compared to the same doses at 2 and 6h. Also, non-progressive motile sperms significantly decreased in E2 group compared to the other groups (P<0.05). Moreover, both patterns C and B were substantially improved in all experimental groups especially in E2 group (P<0.05). Conclusion: Our findings support that the supplementation of sperm with db cAMP+IBMX specially for 4h, could be useful for men with asthenozoospermia to improve the success of assisted reproductive technology.
IBMX protects human proximal tubular epithelial cells from hypoxic stress through suppressing hypoxia-inducible factor-1α expression
Hypoxia predisposes renal fibrosis. This study was conducted to identify novel approaches to ameliorate the pathogenic effect of hypoxia. Using human proximal tubular epithelial cells we showed that a pan-phosphodiesterase (PDE) inhibitor, 3-isobutyl-1-methylxanthine (IBMX) dose and time dependently downregulated hypoxia-inducible factor 1α (HIF-1α) mRNA expression, which was further augmented by addition of a transcriptional inhibitor, actinomycin D. IBMX also increased the cellular cyclic adenosine monophosphate (cAMP) level. Luciferase assay showed that blocking of protein kinase A (PKA) using H89 reduced, while 8-Br-cAMP agonized the repression of HIF-1α promoter activity in hypoxic condition. Deletion of cAMP response element binding sites from the HIF-1α promoter abrogated the effect of IBMX. Western blot and immunofluorescent study confirmed that the CoCl2 induced increased HIF-1α protein in whole cell lysate and in nucleus was reduced by the IBMX. Through this process, IBMX attenuated both CoCl2 and hypoxia induced mRNA expressions of two pro-fibrogenic factors, platelet-derived growth factor B and lysyl oxidase. Moreover, IBMX reduced production of a mesenchymal transformation factor, β-catenin; as well as protected against hypoxia induced cell-death. Taken together, our study showed novel evidence that the PDE inhibitor IBMX can downregulate the transcription of HIF-1α, and thus may attenuate hypoxia induced renal fibrosis.
Forskolin and IBMX Induce Neural Transdifferentiation of MSCs Through Downregulation of the NRSF
Neural differentiation of mesenchymal stem cells is a controversial phenomenon, as it would require transdifferentiation across the mesoderm-ectoderm barrier. However, several laboratories have observed that MSCs are able to be induced to express neural characteristics. Previously, we demonstrated that the cAMP-elevating agents, forskolin and IBMX, induced neural-like differentiation of MSCs, including expression of neural markers and increased sensitivity to neurotransmitters. However, due to the broad range of effects that forskolin and IBMX can elicit through the intracellular second messenger, cAMP, a better mechanistic understanding is required. Here, we show that neural induction by forskolin and IBMX is dependent on downregulation of expression of the master transcriptional regulator, neuron restrictive silencer factor (NRSF), and its downstream target genes. Since silencing of NRSF is known to initiate neural differentiation, it suggests that forskolin and IBMX result in transdifferentiation of MSCs into a neural lineage.
Inhibition of germinal vesicle breakdown using IBMX increases microRNA-21 in the porcine oocyte
Background: Germinal vesicle breakdown (GVBD) occurs during oocyte meiotic maturation, a period when transcriptional processes are virtually inactive. Thus, the maturing oocyte is reliant on processes such as post-transcriptional gene regulation (PTGR) to regulate the mRNA and protein repertoire. MicroRNA (miRNA) are a class of functional small RNA that target mRNA to affect their abundance and translational efficiency. Of particular importance is miRNA-21 (MIR21) due to its role in regulating programmed cell death 4 (PDCD4). The objective of this study was to characterize the abundance and regulation of MIR21 in relation to GVBD.
Methods: Oocytes were collected from aspirated porcine tertiary follicles. Relative abundance of mature MIR21 was quantified at 0, 8, 16, 24, 32, and 42 h of in vitro (IVM) with or without treatment with 3-isobutyl-1-methylxanthine (IBMX).
Results: IBMX increased abundance of MIR21 at 24 h approximately 30-fold compared to control oocytes (P < 0.05), and the induced increase in MIR21 abundance at 24 h was concomitant with premature depletion of PDCD4 protein abundance. To characterize the effect of artificially increasing MIR21 on oocyte competence without inhibiting GVBD, a MIR21 mimic, scrambled microRNA negative control, or nuclease free water was micro-injected into denuded oocytes at 21 h of IVM. The maturation rate of oocytes injected with synthetic MIR21 (63.0 ± 7.5%) was higher than oocytes injected with negative controls (P < 0.05).
Conclusions: Inhibition of nuclear meiotic maturation via IBMX significantly increased MIR21 and decreased its target, PDCD4. Injection of a MIR21 mimic increased oocyte maturation rate. Our results indicate MIR21 is active and important during meiotic maturation of the oocyte.
Effects of IBMX on the ERG of the isolated perfused cat eye
Full-field electroretinograms (ERGs) were recorded from isolated cat eyes perfused through the ophthalmociliary artery with isobutylmethylxanthine (IBMX), an inhibitor of cyclic guanosine monophosphate (cyclic GMP) phosphodiesterase. Low doses of IBMX (0.1-0.3 mM) produced decreased rod ERG amplitudes at low stimulus luminances and increased rod ERG amplitudes at high stimulus luminances. A high dose of IBMX (1.0 mM) initially produced the same effect as the low doses and then led to decreased rod ERG amplitudes at all stimulus luminances. Perfusion with IBMX also resulted in elevations in the semi-saturation luminance (sigma), delayed rod a-wave latencies, delayed rod a-wave and b-wave implicit times, and reduced rod a-wave slopes. Eyes perfused with IBMX (1.0 mM) were also found to have elevated levels of retinal cyclic GMP. These effects of IBMX on the rod ERG are considered in the context of previously described ERGs in selected cases of human retinal degeneration.