Rasarfin targets Ras and ARF6 [1]

达泊西汀通过与负责重新摄取这些化学物质的转运蛋白结合，阻断 5-HT、去甲肾上腺素和多巴胺的摄取。 5-HT>去甲肾上腺素>多巴胺是效力的顺序。达泊西汀对 [3H] 多巴胺摄取的 IC50 值为 202 nM，对 [3H] 去甲肾上腺素摄取的 IC50 值为 1720 nM，它通过去甲肾上腺素再摄取转运蛋白抑制。此外，它还抑制 5-HT 再摄取转运蛋白对 [3H]5-HT 的摄取，抑制值为 1.12 nM。 [1]

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1. 浓度为 50 μM 时，可抑制 GPCR 家族成员（AT1R、B2R、β2AR）的激动剂介导内化，该结果通过 BRET 检测和共聚焦显微镜观察证实 [1]
2. 50 μM 浓度下，能强效抑制 GPCR（AT1R、B2R、β2AR）介导的 ERK1/2 信号通路，以及 EGFR 介导的 MAPK 和 Akt 信号通路，经蛋白质印迹法验证 [1]
3. 50 μM 浓度时，通过 GST-Raf1-RBD 下拉实验和 BRET 动力学分析证实，可阻断 AT1R 介导和 EGFR 介导的 Ras 激活 [1]
4. 经 GST-GGA3-PBD 下拉实验和 BRET 动力学研究表明，50 μM 浓度下可抑制 AT1R 介导的 ARF6 激活 [1]
5. 在 mant-GTP 加载实验中，以浓度依赖方式减少 SOS1 或 EDTA 诱导的纯化 H-Ras 核苷酸交换 [1]
6. 可抑制癌细胞增殖[1]

大鼠口服达泊西汀（1-10 mg/kg；每日一次）可减弱睾酮诱导的前列腺增生，并显着抑制睾酮介导的前列腺重量和相对前列腺重量的增加[2]。

1. Ras 激活实验：利用 GST-Raf1-RBD 融合蛋白，从经 Rasarfin（50 μM）处理或未处理的细胞裂解液中下拉 GTP 结合型 Ras。通过蛋白质印迹法定量 Ras-GTP 相对于总 Ras 的含量，评估对 Ras 激活的抑制作用 [1]
2. ARF6 激活实验：采用 GST-GGA3-PBD 融合蛋白，从暴露于 Rasarfin（50 μM）的细胞裂解液中下拉 GTP 结合型 ARF6。通过蛋白质印迹法定量 ARF6-GTP 相对于总 ARF6 的水平，评价对 ARF6 激活的抑制效果 [1]
3. H-Ras 核苷酸交换实验：将纯化的 H-Ras 与不同浓度的 Rasarfin 共同孵育，随后加入 SOS1 或 EDTA 诱导核苷酸交换。每 30 秒检测一次 mant-GTP 荧光强度，持续 30 分钟，以确定对 H-Ras 激活的抑制作用 [1]

1. GPCR 内化 BRET 检测：将 GPCR（AT1R、B2R、β2AR）和内体转运传感器转染至细胞中。加入 Rasarfin（50 μM）或 DMSO 后，加入激动剂（AngII 或相应配体），检测 BRET 信号以量化配体诱导的受体内化程度 [1]
2. β-arrestin 招募 BRET 检测：将 AT1R 和 β-arrestin1/2 构建体转染至细胞中。加入 Rasarfin（50 μM）或 DMSO 后，用 AngII 刺激细胞，记录 BRET 响应以评估药物对 β-arrestin 招募至 AT1R 的影响 [1]
3. 信号蛋白蛋白质印迹检测：用 Rasarfin（50 μM）或 DMSO 处理细胞，随后用 AngII（针对 GPCRs）或 EGF（针对 EGFR）刺激。制备细胞裂解液，通过蛋白质印迹法检测磷酸化 ERK1/2 和 Akt 的水平，以总 ERK1/2 和 Akt 作为内参对照 [1]
4. 细胞定位共聚焦显微镜观察：将 YFP 标记的 AT1R 或 β-arrestin2 转染至细胞中，用 Rasarfin（50 μM）或 DMSO 处理后，用 AngII 刺激。捕获共聚焦图像以观察受体或 β-arrestin2 的内化及定位情况 [1]
5. 小 GTP 酶激活 BRET 动力学检测：将 AT1R 和针对 Ras、ARF、Rho 或 Rac 的特异性 BRET 传感器转染至细胞中。加入 Rasarfin（50 μM）或 DMSO 后，用 AngII 刺激，实时记录 BRET 信号以分析小 GTP 酶激活的动力学特征 [1]

Animal/Disease Models: Adult male Wistar rat [2]
Doses: 1 mg/kg, 5 mg/kg, 10 mg/kg
Route of Administration: po (oral gavage); 1-10 mg/kg; one time/day
Experimental Results: Testosterone injection resumed brought about most of the changes.

Absorption, Distribution and Excretion
Rapidly absorbed.

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Biological Half-Life
Initial half-life of 1-2 hours.

[1]. Dapoxetine, a novel selective serotonin transport inhibitor for the treatment of premature ejaculation. Ther Clin Risk Manag. 2007 Jun;3(2):277-89.

[2]. Dapoxetine attenuates testosterone-induced prostatic hyperplasia in rats by the regulation of inflammatory and apoptotic proteins. Toxicol Appl Pharmacol. 2016 Nov 15;311:52-60.

Dapoxetine is a member of naphthalenes.
Dapoxetine is a selective serotonin reuptake inhibitor, for the treatment of premature ejaculation. In a phase II proof-of-concept study conducted by PPD, dapoxetine demonstrated a statistically significant increase in ejaculatory latency when compared to placebo. Alza submitted a NDA to the FDA for dapoxetine for the treatment of premature ejaculation in December 2004. In October 2005, the company received a FDA Non-Approvable letter from the FDA, at which time they planned to work with regulators to address outstanding questions.

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Drug Indication
For the treatment of premature ejaculation.

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Mechanism of Action
The drug's mechanism of action is thought to be related to inhibition of neuronal reuptake of serotonin and subsequent potentiation of serotonin activity. The central ejaculatory neural circuit comprises spinal and cerebral areas that form a highly interconnected network. The sympathetic, parasympathetic, and somatic spinal centers, under the influence of sensory genital and cerebral stimuli integrated and processed at the spinal cord level, act in synergy to command physiologic events occurring during ejaculation. Experimental evidence indicates that serotonin (5-HT), throughout brain descending pathways, exerts an inhibitory role on ejaculation. To date, three 5-HT receptor subtypes (5-HT(1A), 5-HT(1B), and 5-HT(2C)) have been postulated to mediate 5-HT's modulating activity on ejaculation.

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Pharmacodynamics
Dapoxetine is a selective serotonin reuptake inhibitor currently undergoing trials through Alza (under license from GenuPro, a collaboration between Eli Lilly and PPD). Dapoxetine is a short-acting SSRI drug currently being considered for approval by the Food and Drug Administration (FDA) for the treatment of premature ejaculation in men, which would make it the first drug approved for such treatment. Despite two clinical trials finished in 2006, experts doubt it will be approved by the FDA soon because SSRIs come with undesirable side-effects after long-term use, such as psychiatric problems, dermatological reactions, increase in body weight, lower sex-drive, nausea, headache, upset stomach and weakness, thus not significantly outweighing the benefit of premature ejaculation medication versus the risks. By contrast with SSRIs approved for depression, which take 2 weeks or longer to reach steady-state concentration, dapoxetine has a unique pharmacokinetic profile, with a short time to maximum serum concentration (about 1 h) and rapid elimination (initial half-life of 1-2 h).

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1. Rasarfin is a novel dual small G protein inhibitor identified from a high-throughput screen of ~115,000 small molecules using an endosomal BRET-based assay with AT1R [1]
2. It binds to the SOS-binding domain of Ras, occupying the cavity normally occupied by SOS residues His911 and Lys939, as revealed by in silico docking and molecular dynamics simulations [1]
3. The binding mode of Rasarfin involves interactions with Ras residues via aromatic features, hydrophobic features, hydrogen bond acceptors/donors, and a halogen bond donor [1]
4. It exhibits functional selectivity, as its analogs show varying effects on GPCR internalization, ERK1/2 activation, and H-Ras nucleotide exchange [1]
5. Rasarfin is useful for inhibiting oncogenic cellular responses by targeting both Ras and ARF6 signaling pathways [1]

MOLECULARWEIGHT

305.4134

305.177

119356-77-3

Dapoxetine hydrochloride;129938-20-1;Dapoxetine-d7 hydrochloride;Dapoxetine-d6;1132642-58-0

71353

Typically exists as solid at room temperature

1.1±0.1 g/cm3

454.4±38.0 °C at 760 mmHg

132.6±29.1 °C

0.0±1.1 mmHg at 25°C

1.607

5.13

12.47

0

2

6

23

337

1

O(C1=C([H])C([H])=C([H])C2=C([H])C([H])=C([H])C([H])=C12)C([H])([H])C([H])([H])[C@@]([H])(C1C([H])=C([H])C([H])=C([H])C=1[H])N(C([H])([H])[H])C([H])([H])[H]

USRHYDPUVLEVMC-FQEVSTJZSA-N

InChI=1S/C21H23NO/c1-22(2)20(18-10-4-3-5-11-18)15-16-23-21-14-8-12-17-9-6-7-13-19(17)21/h3-14,20H,15-16H2,1-2H3/t20-/m0/s1

(1S)-N,N-dimethyl-3-naphthalen-1-yloxy-1-phenylpropan-1-amine

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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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。