JIB-04 is a pan-selective Jumonji histone demethylase inihibitor with IC₅₀s of 230, 340, 855, 445, 435, 1100, and 290 nM for JARID1A, JMJD2E, JMJD3, JMJD2A, JMJD2B, JMJD2C, and JMJD2D, respectively.

IC50: 230 nM (JARID1A), 445 nM (JMJD2A), 435 nM (JMJD2B), 1100 nM (JMJD2C), 290 nM (JMJD2D), 340 nM (JMJD2E), 855 nM (JMJD3)^[1]

JIB-04 is consistently selective for cancer vs. normal cells, demonstrated by the higher sensitivity of lung and prostate cancer lines (with IC₅₀ as low as 10 nM) compared to HBECs and PrSCs/PrECs. JIB-04 inhibits cellular Jumonji demethylase activity, and Jumonji levels affect JIB-04 action in cells^[1]. JIB-04 significantly inhibits the proliferation of GB cell lines and stem-enriched cultures. JIB-04 exerts its maximal inhibitory activity against KDM5A, and modulates the expression of genes involved in the control of cancer cell growth and leads to hypermethylation of H3K4. Furthermore, JIB-04 (2500 nM) activates the autophagy and apoptotic pathways and inactivates PI3K. JIB-04 also cooperates with TMZ in killing GB cells^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

JIB-04 results in a significant reduction in cancer-induced death rates in mice, prolonging survival^[1]. JIB-04 (60, 40 and 20 mg/kg, i.p.) reaches bioactive concentration in the brain of the mice. The orthotopic GB xenograft model shows a trend toward longer survival in JIB-04-treated mice with an Hazard Ratio of 0.5^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

308.76

C₁₇H₁₃ClN₄

199596-05-9

Room temperature in continental US; may vary elsewhere.

DMSO : 50 mg/mL (161.94 mM; Need ultrasonic)

Ethanol : 2 mg/mL (6.48 mM; Need ultrasonic)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 0.5% CMC-Na/saline water

  Solubility: 10 mg/mL (32.39 mM); Suspended solution; Need ultrasonic
• 2.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.08 mg/mL (6.74 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (6.74 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 3.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 1.25 mg/mL (4.05 mM); Clear solution

  此方案可获得 ≥ 1.25 mg/mL (4.05 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 12.5 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Wang L, et al. A small molecule modulates Jumonji histone demethylase activity and selectively inhibits cancer growth. Nat Commun, 2013. 4: p. 2035.

  [2]. Banelli B, et al. Small molecules targeting histone demethylase genes (KDMs) inhibit growth of temozolomide-resistant glioblastoma cells. Oncotarget. 2017 Apr 4.

For cell viability assays, cells are plated at 1500-3000 cells/well in 96 well plates and treated the next day with increasing doses of compound over 4 days and their viability assessed by standard MTS assays using Promega’s Cell Titer or Cell Titer-Glo reagents. Absorbance at 490 nm and 650 nm or luminescence is measured by a Spectra Max or a FlouroStar Omega plate reader. Data are normalized to the untreated controls (100% viability). Each cell line is tested in 2-5 independent assays, each containing 4-8 replicates. IC₅₀ values are calculated using DIVISA, a high-throughput software, developed in house, for storing and analyzing drug sensitivity assays. Dose-response curves are plotted using a non-linear regression model and IC₅₀s are determined from the fitted curves. The average IC₅₀ derived from 2-5 independent assays, each containing 4-8 replicates is reported.

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

For xenografts, 4-6 week old female nude mice are housed under standard conditions in a clean facility at UTSW. Two million H358 cells or five million A549 cells are injected subcutaneously and allowed to grow for 2-3 weeks with monitoring. When tumors reach appr 200 mm³, therapy is started in weight and tumor volume matched pairs (n=7 for each treatment group for each cell line). Drug or vehicle is administered by inter-peritoneal injection in 10% DMSO 90% sesame oil 2 to 3 times weekly for 5 weeks at 110 mg/kg to all mice harboring H358 xenografts or 3 times per week by gavage in 12.5% Cremophor EL, 12.5% DMSO as an aqueous suspension at 55 mg/kg to mice harboring A549 xenografts. Tumor volumes are monitored twice weekly by caliper measurements. Animals are weighed and observed during the five weeks of treatment. At the end point, mice are euthanized by CO₂ asphyxiation and cervical dislocation, and blood, tumors and major organs collected and weighed. Paired, unequal variance, one-tailed t-tests are performed across treatment groups using Excel software.

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Wang L, et al. A small molecule modulates Jumonji histone demethylase activity and selectively inhibits cancer growth. Nat Commun, 2013. 4: p. 2035.

  [2]. Banelli B, et al. Small molecules targeting histone demethylase genes (KDMs) inhibit growth of temozolomide-resistant glioblastoma cells. Oncotarget. 2017 Apr 4.