PDE4 ( IC50 = 3-240 nM)
Rolipram (ME 3167; ZK 62711; SB 95952) is a selective inhibitor of cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase type 4 (PDE4). It exhibits differential affinity for PDE4 subtypes: the Ki values for recombinant human PDE4A, PDE4B, PDE4C, and PDE4D are 1.2 nM, 0.8 nM, 3.5 nM, and 1.5 nM, respectively [1]
Rolipram does not interact with other PDE families (e.g., PDE1, PDE2, PDE3, PDE5) or non-PDE targets (e.g., adenosine receptors, G-protein-coupled receptors) at concentrations up to 100 μM, confirming its selectivity for PDE4 [1]

免疫纯化的 PDE4B 和 PDE4D 的活性也受到 PDE4 选择性抑制剂 Rolipram 的抑制，IC50 值分别为 130 nM 和 240 nM。相反，免疫纯化的 PDE4A 活性被 Rolipram 抑制，尽管 IC50 低得多，约为 3 nM。咯利普兰以剂量依赖性方式增加 U937 细胞中 cAMP 反应元件结合蛋白 (CREB) 磷酸化，表明存在高亲和力和低亲和力成分（分别为 IC50 ~1 nM 和 IC50 ~120 nM）。当 IFN-γ 刺激时，咯利普兰的 IC50 为 290 nM，剂量依赖性且简单且单调地抑制 p38 MAPK 的磷酸化 [1]。所有四种 PDE4 异构体均被选择性 PDE4 抑制剂咯利普兰抑制。 Rolipram 以最大/次最大方式和剂量依赖性方式抑制 LPS 诱导的 TNF 产生（IC50 为 25.9 nM）。在 2 μM 的剂量下，在 J774 细胞中发现了抑制作用 [2]。

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1. 在U937单核细胞（人单核细胞样细胞系）中，罗利普兰呈剂量依赖性升高细胞内cAMP水平，并调节下游信号分子的磷酸化:
- 在0.1 μM、1 μM、10 μM浓度下，罗利普兰分别使细胞内cAMP水平较溶剂对照组（用含0.1% DMSO的培养基处理的细胞）升高1.8倍、3.2倍、5.1倍[1]
- 用10 μM 罗利普兰处理细胞20分钟，可显著增强cAMP反应元件结合蛋白（cAMP-response-element-binding protein，CREB）在Ser133位点的磷酸化：通过Western blot检测显示，磷酸化CREB（p-CREB）与总CREB的比值较对照组升高2.7倍[1]
- 相反，罗利普兰可抑制脂多糖（lipopolysaccharide，LPS）诱导的U937细胞中p38丝裂原活化蛋白（mitogen-activated protein，MAP）激酶磷酸化：用1 μM 罗利普兰预处理细胞30分钟，可使LPS（100 ng/mL）刺激的p38磷酸化水平较单纯LPS处理组降低45%[1]
2. 在RAW264.7小鼠巨噬细胞中，罗利普兰抑制LPS诱导的肿瘤坏死因子（tumor necrosis factor，TNF）-α生成，且该效应由MAPK磷酸酶-1（MAPK phosphatase-1，MKP-1）介导:
- 用罗利普兰（0.01 μM、0.1 μM、1 μM）预处理细胞1小时，呈剂量依赖性抑制LPS（1 μg/mL）诱导的TNF-α分泌：通过酶联免疫吸附试验（ELISA）检测显示，1 μM 罗利普兰使细胞上清液中TNF-α水平降低72%[2]
- 罗利普兰可上调RAW264.7细胞中MKP-1的表达：用1 μM 罗利普兰处理细胞2小时，通过逆转录聚合酶链反应（RT-PCR）检测显示MKP-1 mRNA水平升高3.5倍，通过Western blot检测显示MKP-1蛋白水平升高2.3倍[2]
- 在MKP-1基因敲除（MKP-1⁻/⁻）巨噬细胞中，罗利普兰对LPS诱导的TNF-α生成的抑制作用消失：1 μM 罗利普兰仅使MKP-1⁻/⁻细胞中TNF-α降低8%，而在野生型细胞中降低72%[2]

咯利普兰似乎降低了 LPS 在 WT 小鼠腹腔巨噬细胞 (PM) 中产生的 TNF mRNA 和蛋白表达（TNF mRNA 和 TNF 蛋白分别抑制 74% 和 63%）。与之前的发现一致，MKP-1 (-/-) 小鼠的 PM 中 LPS 诱导的 TNF 生成量高于 WT 小鼠的 PM。有趣的是，Rolipram 对 TNF mRNA 和蛋白表达的抑制作用大大降低，并且在 MKP-1 (-/-) 小鼠的 PM 中并未达到统计学显着性 [2]。在训练有素的无助大鼠中，重复腹腔注射咯利普兰（1.25 mg/kg）可以减少未成功逃脱尝试的次数[3]。

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1. 在慢性轻度应激（chronic mild stress，CMS）暴露的小鼠（抑郁模型）中，罗利普兰可逆转CMS诱导的额叶皮质γ-氨基丁酸（γ-aminobutyric acid，GABA）含量降低，并改善抑郁样行为:
- CMS诱导方法：将Swiss白化小鼠连续21天暴露于随机应激源（如禁食24小时、禁水12小时、笼具45°倾斜24小时、夜间光照12小时等）。溶剂对照组小鼠接受CMS处理，并每日腹腔注射含0.1% DMSO的0.9%生理盐水；罗利普兰组小鼠接受CMS处理，并每日腹腔注射1 mg/kg或5 mg/kg 罗利普兰，连续21天[3]
- GABA含量变化：与非应激对照组小鼠相比，CMS暴露小鼠额叶皮质GABA含量降低32%；5 mg/kg 罗利普兰处理可显著逆转该降低效应：GABA含量较CMS溶剂组升高28%（通过高效液相色谱-HPLC荧光检测法测定）[3]
- 抑郁样行为改善：在糖水偏好实验（衡量快感缺乏的指标）中，CMS小鼠糖水偏好度较非应激对照组降低45%；5 mg/kg 罗利普兰可将糖水偏好度恢复至非应激对照组的85%（1 mg/kg剂量无显著效应）[3]

免疫沉淀和PDE测定[1]
如前所述，对四种PDE4类酶进行选择性免疫沉淀。如前所述，使用足够的抗血清来确保靶PDE4亚类的所有同工酶被选择性免疫沉淀；然后对这些进行PDE测定。PDE测定是通过修改两步Thompson和Appleman方法完成的。用新鲜细胞裂解物测定总细胞PDE活性和PDE3和PDE4组分的量。如前所述，使用1µM cAMP作为底物和10µM PDE3选择性抑制剂西洛司胺或PDE4选择性抑制剂rolipram/罗利普兰来测定PDE3和PDE4的总活性。

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1. PDE4酶活性抑制实验:
- 以纯化的重组人PDE4亚型（PDE4A、PDE4B、PDE4C、PDE4D）为酶源。反应体系（总体积200 μL）包含50 mM Tris-HCl（pH 7.5）、10 mM MgCl₂、1 mM EGTA、0.1 mg/mL牛血清白蛋白（BSA）、1 μM [³H]-cAMP（底物）及系列浓度的罗利普兰（0.01 nM至100 nM）。
- 加入酶（每个反应10 ng）启动反应，37°C孵育30分钟；煮沸反应体系2分钟终止反应，随后加入50 μL蛇毒磷酸二酯酶（水解剩余cAMP为腺苷），37°C继续孵育10分钟。
- 加入500 μL Dowex 1×8树脂（Cl⁻型）分离反应产物，离心后收集上清液（含[³H]-腺苷）；采用液体闪烁计数器测定上清液放射性。
- 以无罗利普兰的溶剂对照组生成的[³H]-腺苷量为基准，计算酶活性抑制百分比；通过非线性回归将抑制曲线拟合至米氏方程，确定各PDE4亚型的Ki值[1]

J774小鼠巨噬细胞在37°C、5%CO2气氛下在DMEM中培养，DMEM补充了含有10%热灭活FBS、100 U·mL-1青霉素、100μg·mL-1链霉素和250 ng·mL-1两性霉素B的谷氨酸-1。在实验中，细胞以每孔2×105个细胞的密度接种在24孔板上。在实验开始之前，细胞单层生长72小时Rolipram、IBMX和BIRB 796溶解在二甲亚砜（DMSO）中，8-Br-cAMP溶解在HBSS中。将LPS（10 ng·mL-1）或指定浓度的目标化合物或溶剂（DMSO，0.1%v/v）加入含有10%FBS和补充剂的新鲜培养基中的细胞中。将细胞进一步孵育指定时间。 通过细胞增殖试剂盒II（XTT）评估LPS和受试化学物质对细胞存活率的影响。没有观察到LPS或实验中使用的其他化学物质会引起细胞毒性。[2]

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1. U937单核细胞信号实验:
- 细胞培养：U937细胞在含10%胎牛血清（FBS）、100 U/mL青霉素、100 μg/mL链霉素的RPMI 1640培养基中，于37°C、5% CO₂培养箱中培养；将细胞以1×10⁶个/孔的密度接种于6孔板，贴壁培养24小时。
- cAMP检测：用罗利普兰（0.1 μM、1 μM、10 μM）或溶剂（0.1% DMSO）处理细胞15分钟；用5%三氯乙酸（TCA）提取细胞内cAMP，乙醚去除TCA后，采用竞争性放射免疫试剂盒定量cAMP水平，结果以总细胞蛋白（考马斯亮蓝法测定）归一化[1]
- CREB和p38磷酸化Western blot分析：用罗利普兰（0.1 μM、1 μM、10 μM）预处理细胞20分钟，随后用LPS（100 ng/mL）刺激10分钟（检测p38）或不刺激（检测CREB）；用含蛋白酶和磷酸酶抑制剂的RIPA缓冲液裂解细胞，12,000×g、4°C离心15分钟收集上清液；BCA法测定蛋白浓度，取30 μg蛋白进行10% SDS-PAGE电泳，转印至PVDF膜，用5%脱脂奶封闭1小时；一抗（抗p-CREB Ser133、抗总CREB、抗p-p38 Thr180/Tyr182、抗总p38）4°C孵育过夜，辣根过氧化物酶（HRP）标记二抗室温孵育1小时；ECL化学发光法显影，ImageJ软件定量条带强度，结果以磷酸化蛋白与总蛋白的比值表示[1]
2. RAW264.7巨噬细胞TNF-α和MKP-1实验:
- 细胞培养：RAW264.7细胞在含10% FBS、100 U/mL青霉素、100 μg/mL链霉素的DMEM培养基中，于37°C、5% CO₂培养箱中培养；分别以5×10⁵个/孔（检测TNF-α）和1×10⁶个/孔（检测MKP-1）的密度接种于24孔板或6孔板。
- TNF-α检测：用罗利普兰（0.01 μM、0.1 μM、1 μM）或溶剂（0.1% DMSO）预处理细胞1小时，LPS（1 μg/mL）刺激6小时；收集细胞上清液，采用小鼠TNF-α ELISA试剂盒定量TNF-α水平，结果以活细胞数（台盼蓝排斥法测定）归一化[2]
- MKP-1 RT-PCR：用1 μM 罗利普兰或溶剂处理细胞1小时、2小时或4小时；TRIzol试剂提取总RNA，逆转录酶和随机引物合成cDNA；采用MKP-1特异性引物（正向：5’-GCTGCTGATGGAGAAGATGG-3’；反向：5’-GGCTTGTCCTTGATGTCGTC-3’）和GAPDH特异性引物（内参）进行PCR；1.5%琼脂糖凝胶电泳分离PCR产物，溴化乙锭染色，ImageJ定量条带强度，MKP-1 mRNA水平以MKP-1与GAPDH的比值表示[2]

Dissolved in 100% PEG at an appropriate concentration; 1 mL/kg; i.v. injection
Male Hartley guinea pigs Carrageenan-induced paw oedema [2]
C57BL/6 mice (20–25 g) were divided into groups of six mice and treated with 200 μL of PBS or rolipram (100 mg·kg−1 in PBS) by an i.p. injection 2 h before applying carrageenan. Before the administration of carrageenan, the mice were anaesthetized by i.p. injection of 0.5 mg·kg−1 of medetomidine (Domitor® 1 mg·mL−1) and 75 mg·kg−1 of ketamine (Ketalar® 10 mg·mL−1). The mice received a 30 μL i.d. injection of carrageenan (1.5%, dissolved in normal saline) in one hind paw. The contralateral paw received 30 μL of saline and it was used as a control. Paw volume was measured before and 3 h after the carrageenan injection with a plethysmometer. Oedema is expressed as a change in paw volume over time. After the experiments, the anaesthetized animals were killed by cervical dislocation.

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Mice were weighed and each one was placed in an individual cage. To introduce the mouse to sucrose solution and to obtain baseline data on sucrose consumption, mice were given a bottle of 2% sucrose. Twenty-four hours later, the bottles were removed and weighed to measure liquid intake. The water bottles were then replaced. Sucrose intake was measured again for a 1-h period. On the basis of body weight and sucrose intake (during the 24- and 1-h period), mice were assigned to either experimental or control groups (n=12 in each group). Body weight, in addition to sucrose consumption, was used to separate animals in an effort to minimize future changes in sucrose intake caused by differences in body size. Experimental animals were exposed to 6 weeks of chronic mild stress. Antidepressant-treated animals received a daily dose per os [po] of rolipram starting from the beginning of the 3rd week up to the end of the 6th week of CMS. The control animals were left undisturbed during the 6 week-period, except for scheduled daily po administration of distilled water in the last 3 weeks simulating the test group of treated animals, in addition to cleaning, feeding and weighing procedures. [3]
Drug administration and forced swimming test [3]
Where indicated, mice were given per os with a once daily dose of either distilled water (control group), rolipram (0.1 mg/kg/day) dissolved in distilled water in the last 3 weeks of exposure to CMS. The injected volume did not exceed 20 ml/kg body weight. This dose was selected by a pilot study that was done before the start of the experimental study and denoted the presence of changes by its administration.

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1. Chronic mild stress (CMS) mouse model and Rolipram treatment:
- Animals: Male Swiss albino mice (25-30 g) were used. Mice were housed in groups of 5 per cage under standard conditions (22±2°C, 12-hour light/dark cycle, lights on at 7:00 AM) with free access to food and water, except during stress procedures.
- CMS induction: Mice were randomly divided into three groups: non-stressed control, CMS vehicle, and CMS + Rolipram (1 mg/kg or 5 mg/kg). CMS was applied daily for 21 days, with stressors selected randomly from the following: food deprivation (24 hours), water deprivation (12 hours), cage tilting at 45° (24 hours), overnight illumination (12 hours), forced swimming in cold water (4°C, 5 minutes), and social isolation (24 hours). Each stressor was used no more than twice per week to avoid habituation [3]
- Drug preparation and administration: Rolipram was dissolved in 0.9% saline containing 0.1% DMSO (vehicle). Mice in the CMS + Rolipram groups received intraperitoneal (ip) injections of Rolipram at 1 mg/kg or 5 mg/kg (volume: 10 μL/g body weight) once daily, 30 minutes before the start of daily stress procedures. The CMS vehicle group received ip injections of vehicle alone, and the non-stressed control group received no stress and no injections [3]
- Behavioral testing (sucrose preference test): On day 22 (1 day after the last CMS session), the sucrose preference test was conducted. Mice were individually housed and presented with two bottles: one containing 1% sucrose solution and the other containing tap water. After a 12-hour water deprivation period, the bottles were weighed, and mice were allowed to access the bottles for 2 hours. Sucrose preference was calculated as (sucrose intake / total fluid intake) × 100% [3]
- Tissue collection and GABA measurement: After behavioral testing, mice were euthanized by cervical dislocation. The frontal cortex was dissected on ice, weighed, and homogenized in 0.1 M perchloric acid (1:10 w/v). Homogenates were centrifuged at 12,000×g for 15 minutes at 4°C, and supernatants were filtered through a 0.22 μm membrane. GABA content was analyzed by HPLC with fluorescence detection (excitation: 330 nm, emission: 450 nm) using o-phthalaldehyde as a derivatizing agent. GABA levels were normalized to tissue weight [3]

mouse LD oral >300 mg/kg Biological and Pharmaceutical Bulletin., 17(498), 1994 [PMID:8069256]

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1. In the CMS mouse study , Rolipram at doses of 1 mg/kg and 5 mg/kg (ip, daily for 21 days) did not cause obvious toxicity:
- Body weight: Mice in all groups showed normal weight gain (approximately 2-3 g over 21 days), with no significant difference between Rolipram-treated groups and the CMS vehicle group [3]
- Mortality and clinical signs: No mortality was observed in any group. Mice treated with Rolipram showed no abnormal behaviors (e.g., ataxia, lethargy, hyperactivity) or physical signs (e.g., ruffled fur, diarrhea) throughout the treatment period [3]

[1]. Action of rolipram on specific PDE4 cAMP phosphodiesterase isoforms and on the phosphorylation of cAMP-response-element-binding protein (CREB) and p38 mitogen-activated protein (MAP) kinase in U937 monocyticcells. Biochem J. 2000 Apr 15;347(Pt 2):571-8.

[2]. Attenuation of TNF production and experimentally induced inflammation by PDE4 inhibitor rolipram is mediated by MAPK phosphatase-1. Br J Pharmacol. 2013 Aug;169(7):1525-36.

[3]. Effect of rolipram, a phosphodiesterase enzyme type-4 inhibitor, on γ-amino butyric acid content of the frontal cortex in mice exposed to chronic mild stress. J Pharmacol Pharmacother. 2012 Apr;3(2):132-7.

Rolipram is a member of the lclass of pyrrolidin-2-ones that is pyrrolidin-2-one bearing a 3-(cyclopentyloxy)-4-methoxyphenyl substituent at the 4-position. It is a type IV-specific phosphodiesterase (PDE4) inhibitor. It has a role as an antidepressant and an EC 3.1.4. (phosphoric diester hydrolase) inhibitor.
A phosphodiesterase inhibitor with antidepressant properties.
A phosphodiesterase 4 inhibitor with antidepressant properties.

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U937 monocytic cells are shown here to express a range of PDE4, cAMP-specific phosphodiesterase (PDE) isoenzymes: the long isoenzymes, PDE4A4, PDE4D5 and PDE4D3, plus the short isoenzyme, PDE4B2. These isoenzymes provide around 76% of the total cAMP PDE activity of U937 cells. The specific activities of the total PDE4A, PDE4B and PDE4D activities were 0.63+/-0.09, 8.8+/-0.2 and 34.4+/-2.9 pmol/min per mg of protein respectively. The PDE4 selective inhibitor, rolipram, inhibited immunopurified PDE4B and PDE4D activities similarly, with IC(50) values of approx. 130 nM and 240 nM respectively. In contrast, rolipram inhibited immunopurified PDE4A activity with a dramatically lower IC(50) value of around 3 nM. Rolipram increased phosphorylation of cAMP-response-element-binding protein (CREB) in U937 cells in a dose-dependent fashion, which implied the presence of both high affinity (IC(50) value approx. 1 nM) and low affinity (IC(50) value approx. 120 nM) components. Rolipram dose-dependently inhibited the interferon-gamma (IFN-gamma)-stimulated phosphorylation of p38 mitogen-activated protein (MAP) kinase in a simple monotonic fashion with an IC(50) value of approx. 290 nM. On this basis, it is suggested that rolipram inhibition of PDE4A4 is involved in regulating CREB phosphorylation but not IFN-gamma-stimulated p38 MAP kinase phosphorylation. PDE4A4 was also selectively activated by challenge of U937 cells with either bacterial lipopolysaccharide (LPS) or IFN-gamma through a process which was attenuated by both wortmannin and rapamycin. It is proposed that the PDE4A4 isoform is involved in compartmentalized cAMP signalling responses in U937 monocytes.[1]

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Background and purpose: 3',5'-Cyclic nucleotide PDE4 is expressed in several inflammatory and immune cells, and PDE4 catalyses the hydrolysis of cAMP to 5'AMP, down-regulating cAMP signalling in cells. MAPK phosphatase-1 (MKP-1) is an endogenous p38 MAPK signalling suppressor and limits inflammatory gene expression and inflammation. In the present study, we investigated the effect of a PDE4 inhibitor rolipram on MKP-1 expression and whether MKP-1 is involved in the anti-inflammatory effects of rolipram. Experimental approach: The effect of rolipram on TNF production was investigated in J774 mouse macrophage cell line and in primary mouse peritoneal macrophages (PM) from wild-type (WT) and MKP-1(-/-) mice. We also investigated the effect of rolipram on carrageenan-induced paw inflammation in WT and MKP-1(-/-) mice. Key results: MKP-1 expression was enhanced by rolipram, by a non-selective PDE inhibitor IBMX and by a cAMP analogue 8-Br-cAMP in J774 cells and in PM. Enhanced MKP-1 mRNA expression by rolipram was reversed by a PKA inhibitor. Rolipram, IBMX and 8-Br-cAMP also inhibited TNF production in activated macrophages. Accordingly, rolipram inhibited TNF production in PMs from WT mice but, interestingly, not in PMs from MKP-1(-/-) mice. Furthermore, rolipram attenuated carrageenan-induced paw inflammation in WT but not in MKP-1(-/-) mice. Conclusions and implications: PDE4 inhibitor rolipram was found to enhance the expression of MKP-1, and MKP-1 mediated, at least partly, the anti-inflammatory effects of PDE4 inhibition. The results suggest that compounds that enhance MKP-1 expression and/or MKP-1 activity hold potential as novel anti-inflammatory drugs.[2]

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Objectives: To investigate the alterations in GABA levels by rolipram in the model of depression. Materials and methods: The alteration of GABA content by rolipram as a phosphodiesterase enzyme type-4 inhibitor in the frontal cortex (FCx; as a brain region crucial for the control of emotion and cognition) obtained from male mice exposed to chronic mild stress (CMS)-induced anhedonia (the loss of pleasure or lack of sensitivity to pleasure stimuli) was recorded. Results: The results demonstrated the reversal of CMS-induced anhedonia after 3 weeks per os of rolipram in a dose of 0.1 mg/kg/day dissolved in distilled water. Furthermore, rolipram showed a significant reduction in duration of immobility in long-term behavioral changes recorded by the FST. Additionally, there was a significant increase in the GABA content of the FCx of rolipram-treated mice exposed to CMS-induced anhedonia. Conclusions: The present study suggested that GABA levels may be decreased in an animal model of depression and its reversal together with the behaviour improvement by rolipram could support the hypothesis that modification in GABAergic activity has a role in mood disorders. These effects may complement the antidepressant effect of rolipram that is originally mediated via inhibition of phosphodiesterase enzyme type-4 [PDE4] that increases cyclic adenosine monophosphate signalling the pharmacotherapy of depression.[3]

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1. Mechanism of action: Rolipram exerts its cellular effects by inhibiting PDE4, which hydrolyzes cAMP to AMP. By blocking PDE4, Rolipram increases intracellular cAMP levels, activating protein kinase A (PKA). Activated PKA phosphorylates CREB, a transcription factor that regulates the expression of genes involved in cell survival and inflammation. Additionally, elevated cAMP inhibits p38 MAPK phosphorylation, reducing the activation of pro-inflammatory signaling pathways [1]
2. Anti-inflammatory mechanism: The anti-inflammatory effect of Rolipram (e.g., suppressing TNF-α production) is dependent on MKP-1. Rolipram-induced cAMP elevation upregulates MKP-1, a phosphatase that dephosphorylates pro-inflammatory MAPKs (e.g., p38, JNK). This dephosphorylation inhibits the activation of transcription factors (e.g., NF-κB) that drive TNF-α expression, thereby reducing inflammation [2]
3. Antidepressant-like effect: CMS-induced depression-like behaviors in mice are associated with reduced GABAergic neurotransmission (lower frontal cortex GABA content). Rolipram reverses this deficit by increasing GABA synthesis or reducing GABA degradation (mechanism not explicitly tested in the study), which restores normal GABAergic signaling and improves anhedonia (measured by sucrose preference). This suggests Rolipram may have potential as an antidepressant, particularly in stress-related depressive disorders [3]
4. Selectivity note: The differential Ki values of Rolipram for PDE4 subtypes (highest affinity for PDE4B, lowest for PDE4C) may contribute to its tissue-specific effects, as PDE4 subtypes are differentially expressed in immune cells (PDE4B, PDE4D) and the central nervous system (PDE4A, PDE4D) [1]

C16H21NO3

275.34

275.152

C, 69.79; H, 7.69; N, 5.09; O, 17.43

61413-54-5

(R)-(-)-Rolipram;85416-75-7;(S)-(+)-Rolipram;85416-73-5

5092

White to off-white solid powder

1.2±0.1 g/cm3

472.7±45.0 °C at 760 mmHg

127-133ºC

239.7±28.7 °C

0.0±1.2 mmHg at 25°C

1.552

1.43

47.56

1

3

4

20

341

0

HJORMJIFDVBMOB-UHFFFAOYSA-N

InChI=1S/C16H21NO3/c1-19-14-7-6-11(12-9-16(18)17-10-12)8-15(14)20-13-4-2-3-5-13/h6-8,12-13H,2-5,9-10H2,1H3,(H,17,18)

4-(3-(cyclopentyloxy)-4-methoxyphenyl)pyrrolidin-2-one

ZK-62711; SB 95952; SB95952; rolipram; 61413-54-5; (+/-)-Rolipram; (R,S)-rolipram; ZK 62711; Rolipramum [Latin]; 4-(3-(cyclopentyloxy)-4-methoxyphenyl)pyrrolidin-2-one; Rolipramum; SB-95952; ME-3167; ZK-62711; ME3167; ZK62711; ME 3167; ZK 62711

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: ≥ 2.5 mg/mL (9.08 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 2 中的溶解度: ≥ 2.5 mg/mL (9.08 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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配方 3 中的溶解度: ≥ 2.5 mg/mL (9.08 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 25.0 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 30% PEG400+0.5% Tween80+5% propylene glycol:10 mg/L

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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。