IL-12/IL-23

Apilimod 的 IC50 约为 20 nM，可抑制人 PBMC 中 IFN-γ/SAC 或 SAC 诱导的 IFN-γ 产生。对高浓度人PBMC中IFN-γ/SAC产生的TNF-α和ConA引起的IL-5，Apimod表现出一定的抑制作用；然而，在所有检测的培养物中，它对 IL-1β、IL-2、IL-4、IL-8 和 IL-18 均表现出一定程度的抑制作用。 IFN-γ/LPS 或 IFN-γ/SAC 刺激大大增加了 p35 和 p40 启动子驱动的荧光素酶活性，而 100 nM Apilimod 完全阻断它 [1]。

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Apilimod (STA-5326)显著抑制IL-12和IL-23的产生[1]
IL-12在先天性和适应性免疫反应中起着不可或缺的作用。IFN-γ是IL-12产生的一种强而选择性的增强剂，只有在LPS或SAC刺激前用IFN-γ延长治疗至少8小时后，这种作用才明显。31这表明，在Th1介导的慢性自身免疫或免疫性疾病中，以反复高水平IFN-γ为特征，IL-12家族的产生是由IFN-γ介导的。为了发现有效的IL-12抑制剂，我们使用IFN-γ/LPS刺激的人外周血单个核细胞进行了表型筛选试验，并筛选了80000个化合物库。通过筛选发现了一种新化合物，即1,3,5-三嗪衍生物。主要优化过程产生并评估了约500种化合物，并发现了独特的吗啉嘧啶衍生物STA-5326。STA-5326强烈抑制IFN-γ/LPS刺激的人PBMC培养物中IL-12的产生，IC50为10 nM（表1）。在IFN-γ/SAC刺激的人外周血单个核细胞中，抑制活性更为明显，其中STA-5326完全抑制IL-12，IC50为1 nM（图1；表1）。即使在10μM的浓度下，也没有观察到细胞存活率的降低。

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Apilimod (STA-5326)在抑制细胞因子产生方面的选择性[1]
细胞因子抑制的选择性对于理解作用机制和体内活性特别重要。STA-5326抑制了IFN-γ/SAC或SAC在人PBMCs中诱导的IFN-γ产生，IC50约为20 nM，而当使用抗CD3和抗CD28抗体刺激T细胞时，该化合物对产生没有显著影响，这表明通过抑制IL-12独立于抑制Th1细胞而发生的T细胞受体依赖性IFN-γ的产生不会被该化合物直接抑制（表2）。在IFN-γ/SAC刺激的人外周血单个核细胞中观察到IL-6产生的显著减少，但在IFN-α/LPS刺激的人单核细胞、IFN-γ/SAC刺激的THP-1和小鼠脾细胞中没有观察到（表2；Y.Wada，未发表的数据，2001-2002）。同样，在IL-10的产生中观察到IFN-γ/SAC特异性抑制。STA-5326在高浓度下对IFN-γ/SAC诱导的TNF-α和ConA诱导的人外周血单个核细胞IL-5表现出一定的抑制作用，但在所有测试的培养物中对IL-1β、IL-2、IL-4、IL-8和IL-18没有抑制作用。

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阿匹利莫（STA-5326）对IL-12 p35和p40启动子活性的影响[1]
在证明STA-5326减少IL-12 p70和p40蛋白的产生后，对p35和p40启动子活性进行了研究，以更深入地了解该化合物的作用机制。用DNA构建体瞬时转染小鼠巨噬细胞系RAW264.7，其中p35和p40启动子指导萤光素酶报告基因的表达。然后用小鼠重组IFN-γ刺激细胞，然后在STA-5326或STA-5392（一种与STA-5326结构相关的无活性化合物）存在或不存在的情况下用LPS或SAC刺激细胞。经IFN-γ/LPS或IFN-γ/SAC刺激后，p35和p40启动子驱动的萤光素酶活性显著诱导，并被100 nM STA-5326完全抑制（图2）。结构上密切相关的非活性化合物STA-5392即使在1μM时也没有效果（图2B）。p35和p40启动子介导的萤光素酶表达的抑制与观察到的p70蛋白产生的抑制具有良好的相关性。该结果表明，STA-5326对IL-12和IL-23的抑制发生在转录水平，并且STA-5326抑制IL-12 p35和IL-12/IL-23 p40基因的转录。

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Apilimod (STA-5326)的活性不需要从头合成蛋白质[1]
皮质类固醇、33-36环腺苷酸、37IFN-β、38和IL-1039通过一种需要从头合成蛋白质的机制对细胞因子（包括IL-12）的产生产生抑制作用。为了确定新的蛋白质合成是否参与STA-5326的抑制机制，与CHX对IL-10抑制IL-12的作用相比，评估了蛋白质合成抑制剂CHX对STA-5326抑制IL-12产生的作用。浓度为5μg/mL的CHX将IL-12的产生减少了一半以上。STA-5326在CHX存在的情况下仍能有效抑制IL-12的产生，并且比单独使用CHX治疗更能减少IL-12的生产，表明CHX不会消除STA-5326的抑制活性（图3）。值得注意的是，STA-5326在CHX存在下的IC50估计值比没有CHX的培养物中的IC50低2至3倍。在同一实验中，浓度为10ng/mL的IL-10在没有CHX的情况下显著降低了IL-12的产生，但在CHX存在的情况下完全失去了抑制活性。

apimod（10 mg/kg，口服）不仅在试验期间连续给药时效果良好，而且在第 30 天开始时效果也很好，此时疾病明显可检测到，但尚未达到最严重的程度。只有在Th1模型中，Apimod才显着减少细胞数量；与载体对照相比，平均而言，受到抑制的细胞减少了 51% ± 8%。 Th2 患者对阿莫德治疗没有反应[1]。在 EAU 小鼠中，阿匹莫德（5 或 20 mg/kg）可降低血液 IL-12 p40 水平，而不影响体重。临床和组织学研究表明，口服阿莫德可降低实验性自身免疫性葡萄膜视网膜炎（EAU）的严重程度[2]。

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阿匹利莫/Apilimod (STA-5326)对体内Th1反应的选择性抑制 STA-5326对体外IL-12和IL-23产生的有效和选择性抑制表明，该化合物应显著抑制体内Th1反应，但不抑制Th2反应。我们评估了STA-5326对体内Th1和Th2小鼠模型的影响，这两种模型分别由结核分枝杆菌和弗氏佐剂在C57BL/6小鼠中诱导，以及蛔虫/氢氧化铝和不完全弗氏佐剂分别在BALB/c小鼠中诱导。STA-5326、类固醇泼尼松龙作为阳性对照或载体从免疫接种当天起口服给小鼠，并收集每组的淋巴结细胞，以评估其对体内Th1和Th2反应诱导的影响。图4分别显示了4个Th1和6个Th2实验中细胞数量相对于初始对照的平均百分比。免疫接种后，这些Th1和Th2模型中载体对照组的淋巴结细胞比相应的初始对照组增加了约3倍。在两种模型中，用泼尼松龙治疗的动物均观察到减少，表明Th1和Th2反应均受到不加选择的抑制。相比之下，STA-5326治疗仅在Th1模型中导致细胞数量显著减少，与赋形剂对照相比，平均抑制百分比为51%±8%。STA-5326治疗在Th2环境中没有效果（图4）。 [1]
为了检查小鼠T细胞的分化，然后以每孔相等的细胞数接种淋巴结细胞，并评估抗CD3/CD28抗体刺激的IFN-γ和IL-4的产生，这两种细胞因子分别代表Th1和Th2。C57BL/6小鼠接种M结核疫苗有效地驱动了载体对照中的体内Th1反应，IFN-γ的产生显著增加（图5）。或者，用蛔虫/氢氧化铝对BALB/c小鼠进行免疫接种会引起体内Th2反应，在载体处理组中观察到IL-4的增加。泼尼松龙治疗不仅能阻断Th1模型中IFN-γ的升高，还能适度阻断Th2模型中IL-4的升高。相比之下，阿匹利莫（STA-5326）仅在Th1模型中抑制IFN-γ，与4个单独的Th1实验中的赋形剂对照组相比，平均减少84%±10%。有趣的是，在STA-5326治疗组中，Th2细胞因子IL-4的产生趋于增加，从6个单独的Th2实验来看，与赋形剂对照组相比，平均增加了232%±91%。这些结果清楚地表明，STA-5326的作用是刺激依赖性的，并且对Th1反应具有选择性。

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Apilimod (STA-5326)在炎症性肠病动物模型中的体内抑制活性 最近的人类和动物研究表明，克罗恩病的局部免疫反应主要是Th1，IL-12在疾病的发生和进展中起着关键作用。40-42为了研究Apilimod (STA-5326)在治疗CD方面的潜力，我们在CD4+CD45Rb高T细胞转移模型中测试了口服该化合物。组织学分析显示，载体处理动物的结肠出现炎症性组织病理学变化，如明显的上皮增生、隐窝长度明显增加、黏膜和LP中慢性炎症细胞的显著浸润，包括隐窝微脓肿、明显的反应性异型性和不同程度的杯状细胞耗竭。在口服阿匹利莫（STA-5326）的动物中，这些变化显著减少，结肠保持正常结构（图6A-B）。组织学评分明确区分了接受STA-5326的动物和接受赋形剂治疗的动物，抑制作用呈剂量依赖性，在4 mg/kg剂量下显著降低，在10 mg/kg剂量下抑制作用更强（图6C）。计算出的结肠与体重比与组织学评分一致，显示出STA-5326治疗的剂量依赖性衰减（图6D）。STA-5326不仅在整个实验过程中给药时有效，而且在第30天开始给药时也有效，此时疾病明显可测量但不是最大值（D.Z.，J.C.，未发表的数据，2002年1月）。此外，本研究中对小鼠来源的LPMC细胞因子的离体分析表明，载体对照的LP细胞产生了增强的IFN-γ水平和检测不到的IL-4水平。STA-5326动物体内IFN-γ的产生大大减少，表明口服STA-5326下调了该炎症性肠病动物模型中的体内Th1反应（图6E）。

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用Apilimod（STA-5326）治疗的小鼠血清中IL-12 p40水平降低。口服5mg/kg或20mg/kg的Apilimod (STA-5326)可在第14天和第18天减轻EAU的严重程度。此外，组织病理学分析显示，用20mg/kg STA-5326治疗的小鼠EAU严重程度显著降低。尽管通过ELISA分析，STA-5326处理的小鼠引流淋巴结细胞的IFN-γ产生增加，但产生IFN-γ的细胞比例没有显著改变。然而，在STA-5326处理的小鼠中，IL-17的产生和产生IL-17的细胞比例显著降低。此外，在效应期口服STA-5326可减轻EAU的严重程度。 结论：这些结果表明，口服IL-12/IL-23抑制剂Apilimod（STA-5326）可有效抑制EAU模型中的炎症，并减少IL-17产生细胞的扩增。STA-5326可能代表了一种治疗人类难治性葡萄膜炎的新方法[2]。

LPMCs的分离和细胞因子的测定[1]
新鲜获得的结肠在无钙/镁的HBSS中洗涤，并在含有EDTA（0.75 mM）、DTT（1 mM）和抗生素（2.5μg/mL两性霉素、50μg/mL庆大霉素）的HBSS在37°C下孵育两次，持续15分钟。组织在37°C下在含有0.5 mg/mL胶原酶D、0.01 mg/mL DNase I和抗生素的RPMI中消化。然后将固有层（LP）细胞分层在40%至100%的Percoll梯度上，并在40%至100%的界面上分离出富含淋巴细胞的群体作为固有层单核细胞（LPMC）。如上所述，在“体内Th1和Th2反应”下，用抗CD3和抗CD28抗体孵育细胞。24小时后取出上清液，使用ELISA试剂盒评估IFN-γ。

体外试验[1]
使用NycoPrep分离人和猴PBMC。使用RosetteSep纯化人单核细胞，通过用粒细胞-巨噬细胞集落刺激因子（GM-CSF；100ng/mL）和IL-4（20ng/mL）培养9天，将树突状细胞从单核细胞中分化出来。通过表型分析，单核细胞和树突状细胞的纯度分别超过70%的CD14+和CD1a+。使用OptiPrep分离小鼠PBMC，并使用ACK裂解缓冲液制备脾细胞。人PBMCs、THP-1细胞和猴PBMCs用人IFN-γ（400 U/mL）预处理22小时，然后用1μg/mL LPS或0.025%SAC刺激18小时。为了研究从头合成的要求，在SAC之前2小时或之前加入环己酰亚胺（CHX）。人外周血单个核细胞也受到抗CD3（0.2μg/mL）和抗CD28（1μg/mL）抗体或ConA（1μg/mL）的刺激。用小鼠IFN-γ（100ng/mL）和SAC（0.05%）的组合刺激小鼠细胞22小时Apilimod（STA-5326）在DMSO中制备；将所有培养物（包括无化合物对照）中的最终DMSO浓度调节至0.25%。
使用抗IL-23 p19多克隆抗体和生物素化山羊抗人p40抗体检测IL-23，并使用重组IL-23计算。使用p40 ELISA试剂盒测定总p40蛋白。使用人IL-12 p70酶联免疫吸附试验（ELISA）试剂盒测量猴IL-12 p7。使用Quantikine HS ELISA试剂盒测量SAC单独诱导的IL-12。使用ELISA试剂盒或Bio-Plex测定法测量其他细胞因子。

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IL-12启动子驱动的萤光素酶测定[1]
为了构建人IL-12 p35和p40启动子/萤光素酶报告子构建体，我们通过聚合酶链式反应（PCR）从人外周血单个核细胞中获得的基因组DNA产生了p35（-1.5kb/+3bp）和p40（-1.3kb/+56bp）启动子片段。将得到的PCR产物连接到pGL3-Basic载体中萤光素酶基因的上游。所有构建体均通过DNA测序进行验证。使用SuperFect转染试剂瞬时转染RAW267.4细胞。用IFN-γ（100 ng/mL）刺激细胞10小时，然后在有或没有测试化合物的情况下用LPS（1μg/mL）刺激细胞16小时。用pCMVβ载体共转染细胞以监测转染效率。根据萤光素酶检测系统和发光β-gal检测系统测定萤光素酶和β-半乳糖苷酶活性。然后使用β-半乳糖苷酶值对萤光素酶活性进行归一化。

CD4+CD45Rbhigh T-cell transfer SCID mouse inflammatory bowel disease [1]
CD4+ T cells in spleen cells from female BALB/c mice were negatively selected using antibodies against B220 (RA3-6B2), CD11b (M1/70), and CD8α (53-6.72), and labeled with FITC-conjugated anti-CD45RB (16A) and PE-conjugated anti-CD4. CD4+ CD45RBhigh cells were defined as the upper 40% of CD45Rb-staining CD4+ cells and were sorted by flow cytometry. Harvested cells were intraperitoneally injected into female C.B-17 SCID mice with 4 × 105 cells per mouse. Apilimod (STA-5326) and vehicle were orally administered once a day, 5 days per week, starting the day following the transfer.
Colon tissues were fixed in 10% buffered formalin and embedded in paraffin. Sections (4 μm) from the ascending, transverse, and descending colon were stained with hematoxylin and eosin. Digital photomicrographs magnified at 40× and 200× of the original from the most affected areas were used for analysis. The extent of colonic inflammation was graded in a blind fashion on a scale of 0 to 3 in each of 4 histologic criteria: crypt elongation, inflammatory-cell infiltration, the number of crypt abscesses, and goblet-cell depletion. The controls were considered to be at baseline with a score of 0. Scores of 1, 2, and 3 represented the following: 2- to 3-fold, 4- to 6-fold, and more than 6-fold increase for crypt elongation; mild, moderate, and severe for inflammatory-cell infiltration; 1 to 2, 3 to 5, and more than 5 colonic crypt abscesses per section; and mild, moderate, and severe for goblet-cell depletion associated with epithelial hyperplasia or inflammatory infiltration. The total score for each animal was the sum of the average scores of ascending, transverse, and descending colon sections in all 4 categories.

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Oral administration of Apilimod (STA-5326) [2]
In most experiments, 5 mg/kg or 20 mg/kg Apilimod (STA-5326) or vehicle only (0.5% carboxyl methyl cellulose) was orally administered once a day for six days a week from day 0 to day 14 after immunization. In the effector phase experiments, 20 mg/kg Apilimod (STA-5326) or vehicle was orally administered once a day, from day 9 to day 14 after immunization.

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IL-12 production in the serum of Apilimod (STA-5326)-treated or vehicle-treated mice after immunization [2]
Mice were immunized as described above, and 5 mg/kg or 20 mg/kg Apilimod (STA-5326) or vehicle alone was orally administered once a day from day 0 to day 14 after immunization. STA-5326-treated or vehicle-treated mice were euthanased on day 18 after immunization, and serum from individual mice were collected for IL-12 p40 measurement using quantikine ELISA kits

Patient characteristics and safety and tolerability of Apilimod (STA-5326).
Twenty-nine eligible patients were included in this trial. The disease characteristics of the study patients were not significantly different between the apilimod- and placebo-treated groups (see Supplementary Table 1, available on the Arthritis & Rheumatism web site at http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1529-0131), with the groups receiving MTX at an average weekly dosage of 21.1 mg and 22.5 mg, respectively. Since we observed favorable safety and pharmacokinetic (MTX and apilimod) profiles in stage 1 (data not shown), the study was continued to stage 2 and subsequently stage 3. In stage 1, 8 of 9 apilimod-treated patients completed the study, while 1 patient who developed side effects (severe headache), withdrew from the study on day 29. The patient refused to undergo the second arthroscopy, although safety and clinical evaluations were completed. All patients treated in stage 2 completed the study. In stage 3 (100 mg twice a day), 3 of 5 apilimod-treated patients continued until day 57, and 1 of these 3 decided to extend the treatment until day 85. Two patients withdrew prior to day 57 due to side effects. In stages 1 and 2, only mild adverse events (mainly gastrointestinal) were observed, in 15 of the 17 patients treated with apilimod (88%). In stage 3, side effects were experienced in all apilimod-treated patients and in patients receiving placebo (Table 1). A detailed listing of the adverse events is provided in Supplementary Table 2, available on the Arthritis & Rheumatism web site at http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1529-0131. [Arthritis Rheum . 2012 Jun;64(6):1750-5]

Objective: To investigate the safety, tolerability, pharmacokinetics, and efficacy of Apilimod (STA-5326) mesylate, an oral interleukin-12 (IL-12)/IL-23 inhibitor, in patients with rheumatoid arthritis (RA).
Methods: We performed a phase IIa, randomized, double-blind, placebo-controlled proof-of-concept study of Apilimod (STA-5326), in combination with methotrexate, in 29 patients with active RA (3:1 ratio of apilimod-treated to placebo-treated patients) in 3 stages. Patients received apilimod 100 mg/day or placebo for 4 weeks (stage 1) or 8 weeks (stage 2). In stage 3, patients received apilimod 100 mg twice a day or placebo for 8 weeks, with an optional extension of 4 weeks. Clinical response (Disease Activity Score in 28 joints [DAS28] and American College of Rheumatology [ACR] criteria) was assessed throughout; synovial tissue samples collected at baseline and on day 29 (stages 1 and 2) or day 57 (stage 3) were stained for cellular markers and cytokines for immunohistochemistry analysis.
Results: While only mild adverse events were observed in stages 1 and 2, in stage 3, all patients experienced headache and/or nausea. Among apilimod-treated patients (100 mg/day), there was a small, but significant, reduction in the DAS28 on day 29 and day 57 compared with baseline. ACR20 response was reached in only 6% of patients on day 29 and 25% of patients on day 57, similar to the percentage of responders in the placebo group. Increasing the dosage (100 mg twice a day) did not improve clinical efficacy. Consistent with clinical results, apilimod did not have an effect on expression of synovial biomarkers. Of importance, we also did not observe an effect of apilimod on synovial IL-12 and IL-23 expression. Conclusion: Our results do not support the notion that IL-12/IL-23 inhibition by apilimod is able to induce robust clinical improvement in RA.
https://pubmed.ncbi.nlm.nih.gov/22170479/

[1]. Selective abrogation of Th1 response by STA-5326, a potent IL-12/IL-23 inhibitor. Blood. 2007 Feb 1;109(3):1156-64.

[3]. Therapeutic effect of the potent IL-12/IL-23 inhibitor STA-5326 on experimental autoimmune uveoretinitis. Arthritis Res Ther. 2008;10(5):R122.

[3]. Billich A. Drug evaluation: apilimod, an oral IL-12/IL-23 inhibitor for the treatment of autoimmune diseases and common variable immunodeficiency.IDrugs. 2007 Jan;10(1):53-9.

Apilimod (STA 5326) is a potent IL-12/IL-23 inhibitor, and strongly inhibits IL-12 in IFNg/SAC-stimulated human PBMCs and SAC-treated monkey PBMCs. Apilimod is a potent and highly selective PIKfyve inhibitor.

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Drug Indication
Investigated for use/treatment in crohn's disease and psoriasis and psoriatic disorders.

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The interleukin-12 (IL-12) cytokine induces the differentiation of naive T cells to the T helper cell type 1 (Th1) phenotype and is integral to the pathogenesis of Th1-mediated immunologic disorders. A more recently discovered IL-12 family member, IL-23, shares the p40 protein subunit with IL-12 and plays a critical role in the generation of effector memory T cells and IL-17-producing T cells. We introduce a novel compound, STA-5326, that down-regulates both IL-12 p35 and IL-12/IL-23 p40 at the transcriptional level, and inhibits the production of both IL-12 and IL-23 cytokines. Oral administration of STA-5326 led to a suppression of the Th1 but not Th2 immune response in mice. In vivo studies using a CD4+CD45Rbhigh T-cell transfer severe combined immunodeficiency (SCID) mouse inflammatory bowel disease model demonstrated that oral administration of STA-5326 markedly reduced inflammatory histopathologic changes in the colon. A striking decrease in interferon-gamma (IFN-gamma) production was observed in ex vivo culture of lamina propria cells harvested from animals treated with STA-5326, indicating a down-regulation of the Th1 response by STA-5326. These results suggest that STA-5326 has potential for use in the treatment of Th1-related autoimmune or immunologic disorders. STA-5326 currently is being evaluated in phase 2 clinical trials in patients with Crohn disease and rheumatoid arthritis. [1]

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STA-5326 is the first potent and selective IL-12/IL-23 inhibitor discovered through screening of a compound library. The unique action of the compound together with the in vivo efficacy in Th1-mediated animal disease models suggest that STA-5326 holds therapeutic promise for the treatment of chronic inflammatory diseases. Injection of monoclonal antibodies recognizing the p40 subunit shared by IL-12 and IL-23 has shown positive results in clinical trials in both patients with psoriasis and patients with Crohn disease. While the antibody acts by neutralizing the IL-12 and IL-23 proteins that already have been produced, STA-5326 acts by selectively shutting off transcription of the p35 and p40 genes. The investigation of the precise mechanism of transcriptional inhibition by STA-5326 is ongoing. STA-5326 now is in clinical evaluation in multiple studies. The compound was shown to be safe in healthy volunteers, and the inhibition of IL-12 production was demonstrated in an ex vivo assay (J.C., unpublished data, 2004). We have completed a phase 2 open-label clinical trial testing STA-5326 in patients with Crohn disease that shows preliminary signs of clinical response by this oral compound. STA-5326 is currently being tested in a blinded and placebo-controlled phase 2b Crohn disease study, and is being evaluated in a phase 2a study in rheumatoid arthritis and common variable immunodeficiency disease (CVID), a disease characterized by elevated IL-12 levels. [1]

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Introduction: The purpose of this study was to determine if oral administration of the interleukin (IL) 12/IL-23 inhibitor, STA-5326, is effective in experimental autoimmune uveoretinitis (EAU). Methods: C57BL/6J mice were immunised with human interphotoreceptor retinoid binding protein peptide (IRBP 1-20). STA-5326 at a dose of either 5 mg/kg or 20 mg/kg, or vehicle alone, was orally administered once a day for six days a week from day 0 to day 14. Fundus examination was performed on day 14 and day 18 after immunisation. Mice were euthanased on day 18 and the eyes were enucleated for histopathological examination. In vivo-primed draining lymph node cells were stimulated with IRBP 1-20 and culture supernatant was harvested for assay of interferon (IFN)-gamma and IL-17 by ELISA. Intracellular expression of IFN-gamma and IL-17 in CD4+ T cells of cultured draining lymph node cells was assessed by flow cytometry. The level of IL-12 p40 in serum was examined in STA-5326-treated or vehicle-treated mice receiving immunisation. [2]
Oral administration of STA-5326 was effective in suppressing inflammation in the EAU model, and reduced the serum level of IL-12/IL-23 p40 and the expansion of IL-17-producing cells. STA-5326 represents a new promising therapeutic modality for refractory uveitis in humans. [2]

C25H34N6O8S2

610.7029

610.187

870087-36-8

Apilimod;541550-19-0; Apilimod mesylate;870087-36-8; 870151-86-3; 1383916-59-3; 870087-37-9 (HCl); 870087-41-5 (besylate)

11527330

White to off-white solid powder

210

3

14

8

41

637

0

CC1=CC(=CC=C1)/C=N/NC2=CC(=NC(=N2)OCCC3=CC=CC=N3)N4CCOCC4.CS(=O)(=O)O.CS(=O)(=O)O

GAJWNIKZLYZYSY-OKUPSQOASA-N

InChI=1S/C23H26N6O2.2CH4O3S/c1-18-5-4-6-19(15-18)17-25-28-21-16-22(29-10-13-30-14-11-29)27-23(26-21)31-12-8-20-7-2-3-9-24-20;2*1-5(2,3)4/h2-7,9,15-17H,8,10-14H2,1H3,(H,26,27,28);2*1H3,(H,2,3,4)/b25-17+;;

(E)-4-(6-(2-(3-methylbenzylidene)hydrazinyl)-2-(2-(pyridin-2-yl)ethoxy)pyrimidin-4-yl)morpholine dimethanesulfonate

LAM-002; LAM 002; STA-5326; STA5326; Apilimod mesylate; 870087-36-8; Apilimod dimesylate; STA 5326 mesylate; STA-5326 mesylate; STA 5326; LAM002; Apilimod mesylate.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~100 mg/mL (~163.75 mM)
DMSO : ~12.5 mg/mL (~20.47 mM)

配方 1 中的溶解度: ≥ 1.25 mg/mL (2.05 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 12.5 mg/mL澄清的DMSO储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入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: ≥ 1.25 mg/mL (2.05 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 12.5 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: ≥ 1.25 mg/mL (2.05 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 12.5 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 100 mg/mL (163.75 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶.

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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。