Go6976 is a Protein Kinase C (PKC) inhibitor, with an IC₅₀ of 20 nM.

IC50: 20 nM (PKC)^[1].

Go6976 is a potent inhibitor of PKC in vitro (IC₅₀ is 20 nM. This compound is structurally related to staurosporine, which is the most potent PKC inhibitor^[1]. UCN-01 is originally identified as a PKC inhibitor. Surprisingly, Go6976 is found to abrogate S and G₂ arrest. Dose-response studies reveal that 30 nM Go6976 is sufficient to cause abrogation of S-phase arrest in 6 h and abrogation of G₂ arrest followed by lethal mitosis in 24 h. Incubation of cells with 100 nM Go6976 is sufficient to cause complete abrogation of S and G₂ arrest at 6 and 24 h, respectively, which is only slightly less potent than in bovine serum. Incubation of cells with UCN-01 or Go6976 alone do not decrease viability compared with control at the concentrations used. Incubation of cells with 5 ng/mL SN38 result in cytostasis, and addition of 50 nM UCN-01 or 100 nM Go6976 to arrested MDA-MB-231 cells cause a dramatic decrease in viable cell number by 96 h^[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.

378.43

C₂₄H₁₈N₄O

136194-77-9

Room temperature in continental US; may vary elsewhere.

Powder	-20°C	3 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : 43.33 mg/mL (114.50 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。

In Vivo:

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% corn oil

  Solubility: ≥ 3.25 mg/mL (8.59 mM); Clear solution

  此方案可获得 ≥ 3.25 mg/mL (8.59 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 32.5 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• 2.

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

  Solubility: 1.39 mg/mL (3.67 mM); Suspended solution; Need ultrasonic

  此方案可获得 1.39 mg/mL (3.67 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 13.9 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% (20% SBE-β-CD in saline)

  Solubility: 1.39 mg/mL (3.67 mM); Suspended solution; Need ultrasonic

  此方案可获得 1.39 mg/mL (3.67 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 13.9 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

• [1]. Qatsha KA, et al. Gö 6976, a selective inhibitor of protein kinase C, is a potent antagonist of human immunodeficiency virus 1 induction from latent/low-level-producing reservoir cells in vitro. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4674-8.

  [2]. Hayamitsu Adachi, et al. Microbial metabolites and derivatives targeted at inflammation and bone diseases therapy: chemistry, biological activity and pharmacology. The Journal of Antibiotics volume 71, pages 60–71 (2018).

  [3]. Mahmoudian S, et al. Influenza A virus proteins PB1 and NS1 are subject to functionally important phosphorylation by protein kinase C. J Gen Virol. 2009;90(Pt 6):1392-1397.

Logarithmically growing cells are incubated with or without 5 ng/mL SN38 for 24 h and then incubated with or without 50 nM UCN-01 or 100 nM Go6976 for the following 24 h. MDA-MB-231 (500 cells) or MCF-10A (1000 cells) are plated in 100 μL in each well of a 96-well plate. The following day, drugs are added at the desired concentrations (Go6976: 1, 3, 10, 30, 100 nM) and with the required schedule to replicate wells (a minimum of 4 wells/concentration). Drugs are removed, and plates are rinsed and then incubated for an additional 6 days. Inhibition of growth was then assessed on the basis of DNA content. Briefly, the media are removed, and attached cells are washed in 0.25×PBS, followed by the addition of 100 μL of H₂O. Cells are lysed by freeze/thawing the plates. Hoechst 33258 is added in high-salt buffer, cells are incubated for 2 h, and fluorescence is measured^[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.

• [1]. Qatsha KA, et al. Gö 6976, a selective inhibitor of protein kinase C, is a potent antagonist of human immunodeficiency virus 1 induction from latent/low-level-producing reservoir cells in vitro. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4674-8.

  [2]. Hayamitsu Adachi, et al. Microbial metabolites and derivatives targeted at inflammation and bone diseases therapy: chemistry, biological activity and pharmacology. The Journal of Antibiotics volume 71, pages 60–71 (2018).

  [3]. Mahmoudian S, et al. Influenza A virus proteins PB1 and NS1 are subject to functionally important phosphorylation by protein kinase C. J Gen Virol. 2009;90(Pt 6):1392-1397.