KJ Pyr 9 is an inhibitor of MYC with a K_d of 6.5 nM in in vitro assay.

Kd: 6.5±1.0 nM (MYC)^[1]

KJ Pyr 9 (KJ-Pyr-9) interferes with MYC-MAX complex formation in the cell, as shown in a protein fragment complementation assay. KJ Pyr 9 specifically inhibits MYC-induced oncogenic transformation in cell culture; it has no or only weak effects on the oncogenic activity of several unrelated oncoproteins. KJ Pyr 9 preferentially interferes with the proliferation of MYC-overexpressing human and avian cells and specifically reduces the MYC-driven transcriptional signature. KJ Pyr 9 against three cell lines is tested known to be dependent on increased MYC activity: NCI-H460, MDA-MB-231, and SUM-159PT. The proliferation of all cell lines tested is inhibited, with IC₅₀ values between 5 and 10 μM. Additionally, the proliferation of Burkitt lymphoma cell lines, which show constitutively high expression of c-MYC, is more sensitive to KJ Pyr 9 (IC₅₀ values between 1 and 2.5 μM)^[1].

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.

To test the in vivo effectiveness of KJ Pyr 9 (KJ-Pyr-9), nude mice receive a xenograft of MDA-MB-231 cells suspended in Matrigel and injected s.c. into the left and right flanks. When the tumors have reached an average volume of 100 mm³, mice are treated daily with 10 mg/kg KJ Pyr 9 or vehicle control by i.p. injection for 31 d. Inhibition of tumor growth by KJ Pyr 9 is noted after 8 d of treatment. By day 31, the tumor volume in the KJ Pyr 9-treated animals has not increased significantly. At the conclusion of the experiment the tumors are extracted and weighed. The weight measurements are in agreement with the volume determinations and confirmed the ability of KJ Pyr 9 to halt tumor growth^[1].

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.

385.37

C₂₂H₁₅N₃O₄

581073-80-5

Room temperature in continental US; may vary elsewhere.

DMSO : 125 mg/mL (324.36 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： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: 2.5 mg/mL (6.49 mM); Suspended solution; Need ultrasonic

  此方案可获得 2.5 mg/mL (6.49 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 2.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (6.49 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (6.49 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 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• 3.

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

  Solubility: 2.08 mg/mL (5.40 mM); Suspended solution; Need ultrasonic

  此方案可获得 2.08 mg/mL (5.40 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  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

• [1]. Hart JR, et al. Inhibitor of MYC identified in a Kröhnke pyridine library. Proc Natl Acad Sci U S A. 2014 Aug 26;111(34):12556-61.

Assays used staining with the redox dye resazurin to measure cell viability. Cells are seeded at 10³ per 100 μL well in 96-well plates and grown in the presence of 2.5% FBS. MDA-MB-231 cells are cultured in DMEM; SUM-159PT cells are cultured in HAM’s F12; and NCI-H460 cells are cultured in RPMI-1640. MDA-MB-231 cells are exposed to KJ Pyr 9 (KJ-Pyr-9) for 216 h with fresh compound-containing medium supplied at 120 and 192 h; SUM-159PT cells are exposed to the compound for 120 h and fresh medium with the appropriate compound concentrations is supplied at 48 h; and NCI-H460 cells are grown with compound for 72 h. Triplicate cultures of P493-6 cells are grown in six-well plates from a starting density of 1×10⁵ cells per mL in 4 mL culture medium per well. Compounds are added immediately following cell seeding. Following compound addition, cells are distributed by vortexing the plate at 400 rpm for 10 s. One hundred-microliter samples are taken after vortexing and counted using a Beckman Coulter Z1 counter at 0, 12, 24, 36, 48, 60, 72, and 96 h of incubation^[1].

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.

Mice^[1]
Ten 8-wk-old female nude mice (HSD:athymic nude-Foxn1^nu) are injected with 5×10⁶ MDA-MB-231 cells s.c. into the left and right flanks. Cells are suspended in high-concentration Matrigel before injection. Xenograft tumors are allowed to grow until the average volume of the tumors reached 100 mm³, as measured by external calipers. At this point, the mice are divided into two groups. One receive 10 mg/kg KJ Pyr 9 and the other receive vehicle only, dosed daily by i.p. injection. Tumor volume and mouse weight are measured daily. Vehicle used in all cases is 10:10:80 Tween 80:DMSO:5% dextrose in water. The mice are treated for a period of 31 d. At the end of the experiment, the mice are euthanized and tumors are excised. Tumors are weighed. Samples of each tumor are fixed in formalin for histology and frozen for Western blotting.

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]. Hart JR, et al. Inhibitor of MYC identified in a Kröhnke pyridine library. Proc Natl Acad Sci U S A. 2014 Aug 26;111(34):12556-61.