Decernotinib (VX-509; VRT-831509; adelatinib)   中文名称: 德塞替尼

JAK3 (Ki = 2.5 nM); JAK1 (Ki = 11 nM); Tyk2 (Ki = 11 nM); JAK2 (Ki = 13 nM); FLT3 (Ki = 1 μM); ROCK I (Ki = 1.5 μM); GSK3β (Ki = 1.8 μM); CDK2/CycA (Ki = 2.6 μM); PknB (Ki = 8 μM)

强 JAK3 抑制剂 decernotinib (VX-509) 对 JAK3、JAK1、JAK2 和 TYK2 的 Ki 值依次为 2.5、11、13 和 11 nM。 decernotinib 的平均 IC50 为 170 ± 101 nM，可有效抑制 T 细胞增殖。它还抑制 IL-2 驱动的 T 细胞增殖（IC50、140 和 400 nM）。在与 CD40L 和 IL-4 发生反应时，VX-509 对 B 细胞也具有细胞毒性（IC50，50 nM）[1]。

在注射胶原蛋白的大鼠中，decernotinib（VX-509、10、25 或 50 mg/kg，口服）显着且剂量依赖性地减少了踝关节直径和爪子重量的增加。在大鼠中，decernotinib 可有效降低骨吸收和软骨退化。 Decernotinib（10、25 或 50 mg/kg，口服，每日两次）可减轻小鼠模型中迟发型超敏反应相关的耳水肿 [1]。

Decernotinib对JAK3活性的影响是通过使用放射测定法测量重组表达的JAK3激酶结构域的残留激酶活性来评估的。实验组分的最终浓度为：100 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM二硫苏糖醇（DTT）， 0.01% BSA, 0.25 nM JAK3, 0.25 mg/mL polyethylene, 5 μM 33P-γ-ATP（200µCi/µmol）。在DMSO中配制10mm的Decernotinib原液，从中制备额外的稀释液。加入底物混合物（100 mM HEPES, 10 mM MgCl2, 0.5 mg/mL聚4y， 10 μM 33P-γ-ATP），与Decernotinib原液混合。该反应通过加入酶混合物[100 mM HEPES (pH 7.5), 10 mM MgCl2, 2 mM DTT, 0.02% BSA， 0.5 nM JAK3]引发。15分钟后，用20%三氯乙酸（TCA）淬灭反应。淬火反应转移到GF/B滤板上，用5% TCA洗涤三次。加入Ultimate Gold闪烁剂（50 μL）后，在Packard TopCount伽马计数器中对样品进行计数。在这个过程中，捕获的放射性是残留JAK3激酶活性的量度。从Decernotinib的活性-浓度滴定曲线来看，Ki值是通过将数据拟合到竞争紧密结合抑制动力学方程来确定的[1]。

健康志愿者全血采集外周血单个核细胞，以1 × 106/mL的密度镀于T75组织培养瓶中。用10 μg/mL植物血凝素在37℃下刺激细胞72小时。72小时后，细胞通过刮刮、洗涤从烧瓶中分离，并以1 × 105/孔的密度镀在96孔板中。加入Decernotinib (9.7 nM ~ 10 μM)， 37℃孵育30分钟，然后用人IL-2刺激。在两行中，只添加DMSO；1行不受IL-2刺激，1行受IL-2刺激作为增殖控制。37℃孵育2天。第2天，细胞用20µCi/mL甲基-[3H]胸腺嘧啶脉冲18-24小时，收集到滤光片上进行射线检测。使用Softmax pro软件分析数据以生成IC50值[1]

The collagen-induced arthritis (CIA) rat model was used to evaluate the effects of oral Decernotinib (VX-509; VRT-831509; adelatinib) [10 mg/kg b.i.d., 25 mg/kg b.i.d., 50 mg/kg b.i.d., 50 mg/kg q.d., or 100 mg/kg q.d.] on joint inflammation and histopathology. Female Lewis rats (157-187 g) are anesthetized with isoflurane and injected with 300 µL Freund’s incomplete adjuvant, containing 2 mg/mL bovine type II collagen, at the base of the tail and two sites on the back on days 0 and 6. The rats are randomized to study groups at the onset of paw swelling (arthritis), which occurs between days 10 and 11. Dosing of either Decernotinib (VX-509; VRT-831509; adelatinib) or vehicle via oral gavage is initiated on the first day of established arthritis and continued to day 6 of arthritis. Dosing volume is 5 mL/kg. Groups are controls (no collagen injection plus vehicle; n = 4), collagen plus vehicle (n = 5), collagen plus Decernotinib (VX-509; VRT-831509; adelatinib) 10 mg/kg b.i.d. (n = 8); collagen plus Decernotinib (VX-509; VRT-831509; adelatinib) 10 mg/kg b.i.d. (n = 8); collagen plus (n = 8); collagen plus (n = 8); collagen plus (n = 8); collagen plu (n = 8); and collagen plus (n = 8); collagen plu (n = 8); all treatments are administered for 6 days. An additional group of rats is given collagen plus 10 mg/kg subcutaneous etanercept, a human tumor necrosis factor-α antagonist, on study days 11 and 14. Caliper measurements of normal (baseline) ankle joints begin on day 9 and continue through the last day of study. Differences in mean ankle diameter are tested for significance using Student’s t test, with significance set at P ≤ 0.05. The rats are euthanized on day 7 of arthritis, which is study day 17 or 18 depending on when animals are randomized to groups; paws and knees are harvested to determine paw weight and to conduct a histopathological analysis of inflammation (knee and ankle), pannus formation (ankle), cartilage destruction (knee), and bone resorption (knee and ankle). Scores range from 0 (normal) to 5 (severe pathology) and are assigned by a veterinary pathologist. Percent inhibition is calculated using the following formula: [(mean of treatment group) − (mean of control)] ÷ [(mean of collagen + vehicle) − (mean of control)]. Kruskal-Wallis one-way analysis of variance nonparametric tests are used to determine statistical significance among the histopathology groups, with significance set at P ≤ 0.05[1].

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Dissolved in 10% vitamin E d-α-tocophenyl polyethylene glycol 1000 succinate and 1% hydroxypropyl methylcellulose acetyl succinate; 50 mg/kg; administrated orally

Collagen-induced arthritis (CIA) rat model

[1]. VX-509 is a potent and selective Janus kinase 3 (JAK3) inhibitor that attenuates inflammation in animal models of autoimmune disease. J Pharmacol Exp Ther. 2015 Mar 11. pii: jpet.114.221176.

Decernotinib has been used in trials studying the treatment of Drug Interactions and Rheumatoid Arthritis.
DECERNOTINIB is a small molecule drug with a maximum clinical trial phase of II and has 1 investigational indication.

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Cytokines, growth factors, and other chemical messengers rely on a class of intracellular nonreceptor tyrosine kinases known as Janus kinases (JAKs) to rapidly transduce intracellular signals. A number of these cytokines are critical for lymphocyte development and mediating immune responses. JAK3 is of particular interest due to its importance in immune function and its expression, which is largely confined to lymphocytes, thus limiting the potential impact of JAK3 inhibition on nonimmune physiology. The aim of this study was to evaluate the potency and selectivity of the investigational JAK3 inhibitor VX-509 (decernotinib) [(R)-2-((2-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-4-yl)amino)-2-methyl-N-(2,2,2-trifluoroethyl)butanamide] against JAK3 kinase activity and inhibition of JAK3-mediated signaling in vitro and JAK3-dependent physiologic processes in vivo. These results demonstrate that VX-509 potently inhibits JAK3 in enzyme assays (Ki = 2.5 nM + 0.7 nM) and cellular assays dependent on JAK3 activity (IC50 range, 50-170 nM), with limited or no measurable potency against other JAK isotypes or non-JAK kinases. VX-509 also showed activity in two animal models of aberrant immune function. VX-509 treatment resulted in dose-dependent reduction in ankle swelling and paw weight and improved paw histopathology scores in the rat collagen-induced arthritis model. In a mouse model of oxazolone-induced delayed-type hypersensitivity, VX-509 reduced the T cell-mediated inflammatory response in skin. These findings demonstrate that VX-509 is a selective and potent inhibitor of JAK3 in vitro and modulates proinflammatory response in models of immune-mediated diseases, such as collagen-induced arthritis and delayed-type hypersensitivity. The data support evaluation of VX-509 for treatment of patients with autoimmune and inflammatory diseases such as rheumatoid arthritis.[1]

C18H19F3N6O

392.38

392.157

C, 55.10; H, 4.88; F, 14.53; N, 21.42; O, 4.08

944842-54-0

59422203

White to off-white solid powder

1.4±0.1 g/cm3

553.6±50.0 °C at 760 mmHg

288.6±30.1 °C

0.0±1.5 mmHg at 25°C

1.603

2.26

99.08

3

8

6

28

548

1

CC[C@](C)(C(=O)NCC(F)(F)F)NC1=NC(=NC=C1)C2=CNC3=C2C=CC=N3

ASUGUQWIHMTFJL-QGZVFWFLSA-N

InChI=1S/C18H19F3N6O/c1-3-17(2,16(28)25-10-18(19,20)21)27-13-6-8-23-15(26-13)12-9-24-14-11(12)5-4-7-22-14/h4-9H,3,10H2,1-2H3,(H,22,24)(H,25,28)(H,23,26,27)/t17-/m1/s1

(R)-2-((2-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-4-yl)amino)-2-methyl-N-(2,2,2-trifluoroethyl)butanamide

PubChem CID 59422203; VX-509; VRT831509 ; 944842-54-0; Adelatinib; Decernotinib (VX-509); Decernotinib(VX-509); VRT-831509; VX509; VX 509 ; VRT 831509 ; Decernotinib; Adelatinib

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： (1). 本产品在运输和储存过程中需避光。  (2). 请将本产品存放在密封且受保护的环境中（例如氮气保护），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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.37 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 (6.37 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 (6.37 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 储备液加入到 900 μL 玉米油中并混合均匀。

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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;
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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。