PDE-4/phosphodiesterase

Apremilast (CC-10004) 的 IC50 为 104 nM (pIC50=6.98±0.2)，可抑制脂多糖 (LPS) 释放 TNF-α。这与 Apremilast 对 PDE4 酶抑制的效力 (IC50=74 nM) 相似，并且几乎完全复制了 Apremilast 先前显示的对外周血单核细胞 (PBMC) 的抑制作用 (IC50=110 nM)。随着细胞内 cAMP 水平的增加，阿普斯特抑制 TNF-α，这些结果令人信服地支持了这一理论。在 PKA、Epac1 或 Epac2 敲低的情况下，未观察到阿普斯特诱导的 IL-10 激活和 TNF-α 抑制]。

当以 5 mg/kg 的剂量口服时，阿普斯特 (CC-10004) 使气囊中产生的 TNF-α 量显着减少 39%（载体的 61±6%，P <0.001），并降低白细胞数量减少 28%（载体的 72±12%，P <0.05）。免疫组织学研究证实，Apremilast 显着减少气囊膜中中性粒细胞的积累。甲氨蝶呤 (MTX) 和阿普斯特均可显着降低小鼠气囊模型中的白细胞浸润，但只有阿普斯特显着抑制 TNF-α 释放。当 MTX (1 mg/kg) 添加到 Apremilast (5 mg/kg) 中时，对白细胞浸润或 TNF-α 释放的抑制并不比单独使用 Apremilast 时更大[1]。已证明新型口服 PDE4 抑制剂阿普斯特可控制炎症介质。口服阿普司特后的平均最大血浆浓度（Cmax）测定为67.00±14.87 ng/mL。阿普司特的血浆浓度迅速下降，最终从血浆中消失，终末半衰期为0.92±0.46 h[2]。

Luminex分析[1]
使用Luminex x-MAP技术对细胞因子和趋化因子进行定量。使用Milliplex多分析物磁珠板分析组织培养上清液和小鼠渗出液中IL-1α、IL-6和IL-10的表达。根据试剂盒方案，使用适当的基质溶液（分别用于上清液和渗出物的培养基或PBS）进行检测。在Luminex 200仪器上收集数据，并使用Analyst 5.1软件进行四参数逻辑斯谛曲线拟合分析。对样品进行了两次化验。由制造商提供的已知参考细胞因子浓度生成的所有标准曲线的R2值计算为或接近1，回收率在80%至120%之间。每个试剂盒都按照预期进行了质量控制。

cAMP测量[1]
用直接cAMP ELISA试剂盒测定细胞内cAMP。将50%融合的Raw 264.7细胞饥饿24小时，以指定浓度的Apremilast (CC-10004)阿普司特 刺激30分钟，然后用脂多糖（LPS）刺激20分钟，并根据制造商的方案分析cAMP。

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TNF-α测定[1]
在96孔板中生长264.7个原始细胞（100000个）。24小时后，用载体（终浓度为0.025%二甲亚砜（DMSO））或指定浓度的阿普司特（CC-10004）刺激细胞。30分钟后，用1μg/ml的LPS刺激细胞4小时。在研究CGS21680、SCH58261、ZM241385、BAY60-6583或GS6201时，在预末用药前15分钟加入腺苷受体配体。在安必利司特前24小时和1小时加入甲氨蝶呤。然后收集上清液，并按照制造商的说明用小鼠TNF-αQuantikine ELISA试剂盒定量TNF-α水平。

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蛋白质印迹[1]
将70%融合的Raw 264.7细胞饥饿24小时，用Apremilast (CC-10004)刺激30分钟，然后用LPS刺激不同时间点（n=4），用放射免疫沉淀法（RIPA）缓冲液裂解细胞，用双辛可宁酸（BCA）测定蛋白质浓度。将蛋白质（4μg）进行7.5或10.0%SDS-PAGE，并转移到硝化纤维膜上。用TBS/Tween-20 0.05-3%BSA阻断非特异性结合。将膜与原代兔多克隆抗pCREB、小鼠单克隆抗CREB、兔多克隆反PDE4和小鼠单克隆抗肌动蛋白（各1:1000）孵育过夜（4°C）。在黑暗中，将膜与山羊抗兔IRDye 800CW 1:10000和山羊抗小鼠IRDye 680 RD 1:10000一起孵育。通过检测近红外荧光的Li cor Odyssey设备对蛋白质进行可视化。由于每种二次抗体都会发出不同光谱的信号，因此与一次抗体孵育同时进行肌动蛋白复制（以检查所有泳道都装载了相同量的蛋白质）。使用Image Studio 2.0.38软件通过密度分析对相应条带的强度进行定量。带强度的变化表示为未受刺激对照的百分比，以尽量减少不同实验之间的差异。

Air pouch model[1]
As previously described, male mice were given weekly intraperitoneal injections of either MTX (1mg/Kg) or vehicle (phosphate-buffered saline; PBS) for 4 weeks. Air pouches were generated by subcutaneous injection of 3 ml of sterile air and reinflated with 1.5 ml of sterile air 2 days later. Vehicle (0.5 % carboxymethylcellulose and 0.25 % Tween 80) or Apremilast (CC-10004) (5 mg/Kg) were orally dosed, with a syringe through a blunt-ended curved feeding tube, 24 h and 1 h before inflammation was induced on day 6 by injection of 1 ml of 2 % carrageenan suspension. Four hours later, mice were killed by CO2 narcosis, and exudates harvested with 2 ml PBS. Leukocytes were counted in a hemocytometer chamber and concentrations of cytokines were measured by ELISA or by the Luminex platform as described below.

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Application to a pharmacokinetic study[2]
Male Sprague Dawley rats (180–220 g) were used to study the pharmacokinetics of Apremilast (CC-10004). Diet was prohibited for 12 h before the experiment, but water was freely available. Blood samples (0.3 mL) were collected from the tail vein into heparinized 1.5 mL polythene tubes at 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8 and 12 h after oral administration of Apremilast (CC-10004) (6.0 mg/kg). The samples were immediately centrifuged at 4,000 g for 8 min. The plasma obtained (100 µL) was stored at −20°C until analysis. Plasma apremilast concentration versus time data for each rat was analyzed by DAS (Drug and statistics) software.

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[1]. Apremilast, a novel phosphodiesterase 4 (PDE4) inhibitor, regulates inflammation through multiple cAMP downstream effectors. Arthritis Res Ther. 2015 Sep 15;17:249.

[2]. Determination of Apremilast in Rat Plasma by UPLC-MS-MS and Its Application to a Pharmacokinetic Study. J Chromatogr Sci. 2016 Sep;54(8):1336-40.

Dissolved in 0.5% carboxymethylcellulose and 0.25% Tween 80; 5 mg/kg; P.O.

Mouse xenograft model of psoriasis (Beige-SCID mice)

Absorption, Distribution and Excretion
Oral apromisit is well absorbed, with an absolute bioavailability of approximately 73%. The time to peak concentration (Tmax) is approximately 2.5 hours, and a pharmacokinetic study reported a peak plasma concentration (Cmax) of approximately 584 ng/mL. Food intake does not appear to affect apromisit absorption. Only 3% and 7% of the apromisit dose were detected unchanged in urine and feces, indicating extensive metabolism and high absorption. The mean apparent volume of distribution (Vd) is approximately 87 L, suggesting that apromisit is distributed in the extravascular space. In healthy patients, the plasma clearance of apromisit is approximately 10 L/hour. The human plasma protein binding of apromisit is approximately 68%. The mean apparent volume of distribution (Vd) is 87 L. This study evaluated milk excretion in lactating CD1 mice following oral administration of apromisit. In this study, female mice approximately 13 days postpartum were given a single oral dose of 10 mg/kg apromiscarriage via gavage (10 mL/kg). Milk and blood samples were collected from five mice at 1, 6, and 24 hours post-administration, and plasma and milk apromiscarriage concentrations were determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS). At 1 and 6 hours post-administration, the average plasma apromiscarriage concentrations were 984 ng/mL and 138 ng/mL, respectively, while the average milk apromiscarriage concentrations were 1441 ng/mL and 186 ng/mL, respectively. The average milk-to-plasma concentration ratio ranged from 1.46 to 1.62, indicating that apromiscarriage can be transferred to mouse milk. At 24 hours, both plasma and milk concentrations were below the detection limit of 3 ng/mL. In monkeys, pregnant animals were administered apremilast orally daily from day 20 to day 50 of gestation, with a single oral dose on day 100 of gestation, at doses of 20, 50, 200, and 1000 mg/kg/day (n = 16 per group at the start of the study). Maternal and fetal blood were collected 5 hours after administration on day 100 of gestation. Fetal to maternal plasma concentration ratios were between 0.3 and 0.4 in all dose groups, indicating that apremilast can cross the monkey placenta. As part of a fertility and developmental toxicity study in female CD1 mice and an embryo-fetal development study in cynomolgus monkeys, the transplacental transport of apremilast was evaluated. In mice, apremilast was administered orally daily from day 15 before cohabitation until day 15 of the presumptive gestation at doses of 10, 20, 40, and 80 mg/kg/day. Blood samples were collected from pregnant mice (n=3 at each time point) at 0.5, 2, 4, 8, and 24 hours after administration on day 15 of gestation. Fetal blood samples were also collected from mice sacrificed 24 hours after administration. The increase in maternal plasma apromisin concentration was less than dose-proportional. Fetal plasma apromisin concentrations varied considerably at 24 hours; in 6 of the 10 litters evaluated, concentrations were below the limit of quantitation (1 ng/mL). Apromisin was detected in fetal plasma from 4 of the 10 litters evaluated, ranging from 14.5 to 2813 ng/mL. The average fetal-to-maternal plasma concentration ratio ranged from 0.3 to 1.07, indicating that apromisin can cross the mouse placenta.
For more complete data on the absorption, distribution, and excretion of apromisin (13 parameters), please visit the HSDB record page.

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Metabolism/Metabolites
Apramistrast is primarily metabolized through multiple pathways, including oxidation, hydrolysis, and conjugation. It has approximately 23 metabolites. CYP3A4 mainly mediates the oxidative metabolism of this drug, while CYP1A2 and CYP2A6 enzymes contribute less. The major metabolite of apramistrast, M12, is an inactive glucuronide conjugate of the O-demethylated drug. Other major metabolites, such as M14 and M16, exhibit significantly lower activity in inhibiting PDE4 and inflammatory mediators than their parent drug, apramistrast. After oral administration, the primary metabolites detected in plasma are parent apramistrast (45%) and the inactive metabolite O-demethylapramistrast glucuronide (39%). Minor metabolites M7 and M17 are active, but their concentrations account for only 2% or less of the total apramistrast concentration, and may contribute little to its effect. In healthy subjects, the clearance of apromiscalcium in plasma is approximately 10 L/hr, with a terminal elimination half-life of approximately 6–9 hours. Following oral administration of radiolabeled apromiscalcium, approximately 58% and 39% of the radioactive material are recovered in urine and feces, respectively, with approximately 3% and 7% of the radioactive dose recovered in the form of apromiscalcium in urine and feces, respectively. Following oral administration in humans, apromiscalcium is the major circulating component (45%), followed by the inactive metabolite M12 (39%), which is an O-demethylated glucuronide conjugate of apromiscalcium. Apromiscalcium is extensively metabolized in humans, with up to 23 metabolites identified in plasma, urine, and feces. The metabolic pathways of apromiscalcium include cytochrome P450 (CYP) oxidative metabolism (followed by glucuronidation) and non-CYP-mediated hydrolysis. In vitro studies have shown that CYP metabolism of apromiscalcium is primarily mediated by CYP3A4, with smaller contributions from CYP1A2 and CYP2A6. In an oral study, the concentrations of total radioactivity (e.g., the parent compound and its metabolites) and the parent compound in plasma were higher in female rats than in male rats. The total radioactivity AUC was 25 to 96 times higher in male rats than in female rats, while the difference was only 2 to 3 times, indicating that male rats metabolized the drug more extensively than female rats. In the same study, after six consecutive days of administration, female mice showed drug accumulation in Cmax and AUC, while no accumulation was observed in male mice. In a bile duct cannulation study of male mice, after a single oral administration of 10 mg/kg (14)C-apromiscalcium, 54% and 16% of the radioactive dose were excreted via bile and urine, respectively, indicating that at least 70% of the radioactive dose was absorbed by the mice, suggesting moderate first-pass metabolism of apromiscalcium. Molecular toxicokinetics showed that exposure increased with increasing dose, but the trend toward increased exposure was less proportional to the dose at doses exceeding 100 mg/kg/day. No sex differences or conversion to its R enantiomer were observed in mice. For more complete data on the metabolism/metabolites of apramistrast (6 metabolites), please visit the HSDB record page. Biological Half-Life: The mean elimination half-life of this drug is 6–9 hours. The terminal elimination half-life is approximately 6–9 hours.

Toxicity Summary
Identification and Use: Apramisartil is a white to pale yellow powder. Apramisartil is used to treat adults with active psoriatic arthritis. It is also used to treat patients with moderate to severe plaque psoriasis who are suitable for phototherapy or systemic therapy. Human Exposure and Toxicity: The most common adverse reactions are diarrhea, nausea, upper respiratory tract infection, and headache, including tension headache. Apramisartil did not induce chromosomal aberrations in cultured human peripheral blood lymphocytes with or without metabolic activation. Animal Studies: Apramisartil has low acute toxicity. Repeated-dose oral toxicity studies lasting up to 6 months in mice (dose levels of 10, 100, and 1000 mg/kg/day), up to 12 months in monkeys (dose levels of 60, 180, and 600 mg/kg/day), and up to 90 days in rats have evaluated apramisartil. Apramisartil-related deaths have been observed in mice and rats, primarily attributable to vascular and/or perivascular inflammation. Dose-related inflammatory responses were observed primarily in mice and rats, including neutropenia, lymphopenia, and changes in serum proteins (decreased albumin, increased globulins, haptoglobin, C-reactive protein (CRP), and/or fibrinogen). These inflammatory responses were associated with arteritis and perivascular inflammation in various tissues and organs (e.g., mesentery, heart, lung, thymus, liver, skeletal muscle, mammary gland, skin, and pancreas) in mice and rats, but were not observed in monkeys, even though the systemic exposure in monkeys was higher than that in mice and rats. Complete or partial reversibility of the inflammatory responses was observed in mice and rats. Other target organs for apramiscut toxicity included non-adverse centrilobular hepatocyte hypertrophy in the liver (mice) and varying degrees of lymphopenia in lymphoid tissues (mice and rats). Long-term studies of apramiscut in mice and rats have been conducted to assess its carcinogenic potential. In mice, no evidence of apromiscalcium-induced tumors was observed at oral doses up to 8.8 times the maximum recommended human dose (MRHD) (based on AUC, i.e., 1000 mg/kg/day). In rats, no evidence of apromiscalcium-induced tumors was observed at oral doses up to approximately 0.08 and 1.1 times the MRHD (20 mg/kg/day for males and 3 mg/kg/day for females, respectively). In a male mouse fertility study, oral doses of apromiscalcium at 1, 10, 25, and 50 mg/kg/day did not affect male fertility. In a combined study of female mouse fertility and embryo-fetal developmental toxicity, oral doses of apromiscalcium at 10, 20, 40, and 80 mg/kg/day showed altered estrous cycles and prolonged mating time starting from 20 mg/kg/day. However, all mice mated successfully, and pregnancy rates were unaffected. Apremilast did not induce mutations in the Ames assay. At doses up to 2000 mg/kg/day, apremilast did not show chromosome breakage induction in the in vivo mouse micronucleus assay.

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Interactions
Otezila has not been evaluated and is not recommended for use in combination with biologics used to treat psoriasis (such as TNF antagonists and anti-IL-12/23 p40 antibodies). Otezila is not recommended for use in combination with these biologics.
Otezila has not been evaluated and is not recommended for use in combination with potent immunosuppressants (such as cyclosporine and tacrolimus). Otezila is not recommended for use in combination with potent immunosuppressants.
When used in combination with the CYP3A4 inducer rifampin, the exposure (AUC) and maximum concentration (Cmax) of apremilast were reduced by 72% and 43%, respectively, which may lead to a decrease in the clinical efficacy of apremilast. Therefore, it is not recommended to use olerazine in combination with rifampin or other CYP3A4 inducers (such as phenobarbital, carbamazepine, phenytoin sodium). St. John's wort is a CYP3A4 inducer, and its combination with olerazine may lead to reduced efficacy or weakened clinical response; therefore, such combination is not recommended.

[1]. Apremilast, a novel phosphodiesterase 4 (PDE4) inhibitor, regulates inflammation through multiple cAMP downstream effectors. Arthritis Res Ther. 2015 Sep 15;17:249.

[2]. Determination of Apremilast in Rat Plasma by UPLC-MS-MS and Its Application to a Pharmacokinetic Study. J Chromatogr Sci. 2016 Sep;54(8):1336-40.

Therapeutic Uses

Nonsteroidal anti-inflammatory drug; Phosphodiesterase inhibitor
Otezla is indicated for the treatment of adult patients with active psoriatic arthritis. /US product label includes/
Otezla is indicated for the treatment of patients with moderate to severe plaque psoriasis who are eligible for phototherapy or systemic therapy. /US product label includes/
Exploratory Treatment/This study aims to/evaluate the efficacy and safety of oral phosphodiesterase 4 inhibitor apremilast for the treatment of ankylosing spondylitis (AS) through a preliminary study monitoring symptoms and signs. This study includes an exploratory investigation of the effects of PDE4 inhibition on bone biological blood biomarkers. In this double-blind, placebo-controlled, single-center phase II study, patients with symptoms of active ankylosing spondylitis (AS) as shown by MRI were randomized to receive apremilast 30 mg twice daily (BID) or placebo for 12 weeks. Bath index was continuously monitored during the study. Patients were followed up for 4 weeks after discontinuation of treatment. Bone biomarkers were assessed at baseline and day 85. A total of 38 participants were randomized, and 36 completed the study. Although the primary endpoint (change in BASDAI score at week 12) was not met, the apremilast group showed greater improvement from baseline on all clinical assessment endpoints compared to the placebo group, with mean changes in BASDAI (-1.59 ± 1.48 vs -0.77 ± 1.47), BASFI (-1.74 ± 1.91 vs -0.28 ± 1.61), and BASMI (-0.51 ± 1.02 vs -0.21 ± 0.67) being significantly greater than in the placebo group; however, these differences did not reach statistical significance. Four weeks after discontinuation of apremilast, all clinical endpoints returned to baseline. Six patients (35.3%) in the apremilast group achieved ASAS20 remission, compared to only three (15.8%) in the placebo group (p=0.25). Serum RANKL, RANKL/osteoplastin ratio, and plasma scleroprotein levels showed statistically significant decreases, but serum DKK-1, bone alkaline phosphatase, TRAP5b, MMP3, osteoprotegerin, or osteocalcin levels showed no significant changes. Although this is a small, preliminary study, these results suggest that apremilast may be effective and well-tolerated in ankylosing spondylitis and can modulate bone biomarkers. These data support further investigation into the role of apremilast in axial inflammation.
Exploring Treatment: Discoid lupus erythematosus (DLE) is a chronic inflammatory disease mediated by Th1 cells. Apremilast is a novel oral PDE4 enzyme inhibitor that blocks the production of IL-12, IL-23, TNF-α, and INF-12 by leukocytes, thereby inhibiting Th1 and Th17-mediated immune responses. It has been shown to have clinical efficacy in psoriasis, as well as rheumatoid arthritis and psoriatic arthritis. In 8 patients with active discoid lupus erythematosus, after 85 days of treatment with apromiscalcium 20 mg twice daily, the disease area and severity index (CLASI) of lupus cutaneum was significantly reduced (P<0.05). Drug-related adverse events were mild and transient. This is the first open-label study of apromiscalcium for the treatment of discoid lupus erythematosus. Our observations suggest that apromiscalcium may be a safe and effective treatment option for discoid lupus erythematosus.

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Drug Warnings
Use of Otezla is associated with an increased risk of adverse reactions related to depression. Prescribing physicians should carefully weigh the risks and benefits of Otezla treatment in patients with a history of depression and/or suicidal ideation or behavior before administering it. Patients, their caregivers, and families should be informed to be alert for the onset or exacerbation of depression, suicidal ideation, or other mood changes, and to contact their healthcare provider immediately if such changes occur. Prescribing physicians should carefully assess the risks and benefits of continuing Otezla treatment if such conditions occur.
The safety and efficacy of octazine in children under 18 years of age have not been established.
It is currently unknown whether octazine or its metabolites are present in human milk; however, apromiscarriage has been detected in the milk of lactating mice. Because many drugs are present in human milk, caution should be exercised when octazine is taken by breastfeeding women.
FDA Pregnancy Risk Category: C/Risk cannot be ruled out. There is a lack of adequate, well-controlled clinical studies, and animal studies have not shown any risk to the fetus or lack relevant data. Taking this drug during pregnancy may cause harm to the fetus; however, the potential benefits may outweigh the potential risks. /
For more complete data on drug warnings for apromiscarriage (11 in total), please visit the HSDB record page.

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Pharmacodynamics
Apramistrastem can reduce, but not completely inhibit, a variety of inflammatory cytokines, such as IL-1α, IL-6, IL-8, IL-10, MCP-1, MIP-1β, MMP-3, and TNF-α, thereby alleviating symptoms of psoriasis and Behcet's disease caused by increased levels of these inflammatory mediators. This drug has also been shown to effectively relieve pain caused by oral ulcers in Behcet's disease. Apramistrastem may cause weight loss and exacerbation of depressive symptoms, and may even trigger suicidal thoughts or behaviors. Monitoring for depressive symptoms is recommended, and patients should seek medical attention promptly if symptoms occur, especially those with a history of depression. The necessity of using apramistrastem and the risk of exacerbation of depression and suicide should be carefully evaluated. If weight loss occurs, the degree of weight loss should be assessed, and the need to discontinue apramistrastem should be considered.

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Introduction: This study aims to elucidate the intracellular signaling pathway of the PDE4 inhibitor apramistrastem and to explore the interaction between apramistrastem, methotrexate, and adenosine A2A receptor (A2AR). Methods: In the Raw264.7 mononuclear cell line, the levels of intracellular cAMP, TNF-α, IL-10, IL-6, and IL-1α were detected after incubation with apremilast and LPS. PKA, Epac1/2 (an intermediate in the cAMP signaling pathway), and A2AR were knocked down by shRNA transfection, and their interactions with A2AR, A2BR, and methotrexate were investigated in vitro and in a mouse air sac model. Statistical differences were determined using one-way or two-way ANOVA or Student's t-test. The nominal α level for all tests was set at 0.05. A p-value <0.05 was considered statistically significant. Results: In vitro experiments showed that apremilast increased intracellular cAMP levels and inhibited TNF-α release (IC50 = 104 nM), while the specific A2AR agonist CGS21680 (1 μM) enhanced the potency of apremilast (IC50 = 25 nM). In this cell line, apramisartine increased IL-10 production. Knockdown of PKA, Epac1, and Epac2 prevented apramisartine's inhibitory effect on TNF-α and its stimulatory effect on IL-10. In a mouse air sac model, both apramisartine and MTX significantly inhibited leukocyte infiltration, while apramisartine (but not MTX) significantly inhibited TNF-α release. Adding methotrexate (MTX, 1 mg/kg) to apramisartine (5 mg/kg) did not enhance its inhibitory effect on leukocyte infiltration or TNF-α release compared to apramisartine alone. Conclusion: The immunomodulatory effects of apramisartine appear to be mediated through cAMP and its downstream effector molecules PKA, Epac1, and Epac2. A2AR agonists enhanced the inhibitory effect of apramisartine on TNF-α, consistent with the cAMP-enhancing effect of this receptor. Since A2AR is also involved in the anti-inflammatory effect of MTX, the mechanisms of action of both drugs involve the cAMP-dependent pathway and are essentially partially overlapping. [1]

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We developed and validated a rapid, sensitive and selective ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method for determining the concentration of apromiscal in rat plasma and for pharmacokinetic studies. Sample preparation was performed using a simple single-step deproteinization method, adding 0.2 mL of acetonitrile to 0.1 mL of plasma sample. Plasma samples were separated by ultra-high performance liquid chromatography (UPLC) using an Acquity UPLC BEH C18 column and a mobile phase of acetonitrile-0.1% formic acid aqueous solution, with gradient elution. The total run time was 3.0 min, and the elution time for apromiscal was 1.27 min. The detection was performed using a triple quadrupole tandem mass spectrometer in multiple reaction monitoring (MRM) mode. The ion pairs for apromisci were m/z 461.3 → 257.1, and for carbamazepine (internal standard) were m/z 237.2 → 194.2. The calibration curves showed a linear relationship in the range of 0.1–100 ng/mL, with a limit of quantitation of 0.1 ng/mL. The average recovery rate of apromisci in plasma was 83.2%–87.5%. The intra-day and inter-day precision were both less than 9.6%. This method has been successfully applied to the pharmacokinetic study of rats after oral administration of 6.0 mg/kg apromisci. [2]

C22H24N2O7S

460.50016

460.13

C, 57.38; H, 5.25; N, 6.08; O, 24.32; S, 6.96

608141-44-2

Apremilast-d5;1258597-47-5; (R)-Apremilast; 608141-44-2;(Rac)-Apremilast-d5; 1258597-61-3; 253168-86-4

9912092

White to yellow solid powder

1.4±0.1 g/cm3

741.3±60.0 °C at 760 mmHg

402.1±32.9 °C

0.0±2.5 mmHg at 25°C

1.612

1.75

127.46

1

7

8

32

825

1

S(C)(C[C@@H](C1C=CC(=C(C=1)OCC)OC)N1C(C2C=CC=C(C=2C1=O)N([H])C(C)=O)=O)(=O)=O

IMOZEMNVLZVGJZ-KRWDZBQOSA-N

InChI=1S/C22H24N2O7S/c1-5-31-19-11-14(9-10-18(19)30-3)17(12-32(4,28)29)24-21(26)15-7-6-8-16(23-13(2)25)20(15)22(24)27/h6-11,17H,5,12H2,1-4H3,(H,23,25)/t17-/m0/s1

N-[2-[(1R)-1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-1,3-dioxoisoindol-4-yl]acetamide

(R)-Apremilast; 608141-44-2; (R)-N-(2-(1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl)-1,3-dioxoisoindolin-4-yl)acetamide; N-[2-[(1R)-1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-1,3-dioxoisoindol-4-yl]acetamide; N-[2-[(1R)-1-(3-Ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl]-2,3-dihydro-1,3-dioxo-1H-isoindol-4-yl]-acetamide;; CHEMBL473204; SCHEMBL7135116;

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)

DMSO : ~100 mg/mL (~217.16 mM)

配方 1 中的溶解度: ≥ 12.5 mg/mL (27.14 mM) (饱和度未知) 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。
例如，若需制备1 mL的工作液，可将100 μL 125.0 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，混合均匀。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (5.43 mM) (饱和度未知) 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。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；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 3 中的溶解度: ≥ 2.5 mg/mL (5.43 mM) (饱和度未知) 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。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 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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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。