Staurosporine (AM-2282)

Biological Activity

Staurosporine (antibiotic AM-2282 or STS) is a therapeutic agent that inhibits tumor cell growth by inducing cell death via intrinsic apoptotic pathways. Staurosporine is a prototypical ATP-competitive kinase inhibitor in that it binds to many kinases with high affinity, though with little selectivity. Staurosporine also inhibits a variety of other protein kinases, including PKA, PKG, phosphorylase kinase, S6 kinase, MLCK, CAM PKII, cdc2, v-Src, Lyn, c-Fgr and Syk with IC50 values of 15 nM, 18 nM, 3 nM, 5 nM, 21 nM, 20 nM, 9 nM, 6 nM, 20 nM, 2 nM and 16 nM respectively. Staurosporine induces apoptosis of human foreskin fibroblasts AG-1518, depending on the lysosomal cathepsins D mediated cytochrome c release and caspase activation. Moreover, Staurosporine triggers a novel intrinsic apoptosis pathway, relying on the activation of caspase-9 in the absence of Apaf-1.

Product Citations

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  Inflamm Regen. 2023 Jun 20;43(1):31.

  Promotion of axon regeneration and protection on injured retinal ganglion cells by rCXCL2
  Staurosporine (AM-2282) purchased from AbMole

• 

  Biomaterials. 2021 Aug;275:120958.

  Tumor progress intercept by intervening in Caveolin-1 related intercellular communication via ROS-sensitive c-Myc targeting therapy
  Staurosporine (AM-2282) purchased from AbMole

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  Pak Vet J. 2021 Mar.

  Evaluation of the PBMC Proliferation, Apoptosis and Cytokines Profiling in Cattle Infected with Mycoplasma bovis Strain 07801
  Staurosporine (AM-2282) purchased from AbMole

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  PLoS One. 2019 Oct 10;14(10):e0223096.

  Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors.
  Staurosporine (AM-2282) purchased from AbMole

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  Food Chem. 2017 Mar 15;219:304-310.

  Effects of protein phosphorylation on color stability of ground meat
  Staurosporine (AM-2282) purchased from AbMole

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  Food Chem. 2017 Sep 20.

  Dephosphorylation enhances postmortem degradation of myofibrillar proteins
  Staurosporine (AM-2282) purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Food Chem (2017). Figure 3. Staurosporine (Abmole Bioscience,Houston, TX, USA)
	Method	Measured myofibril fragmentation index
	Cell Lines	Muscles
	Concentrations	0.144 μmol/5 g
	Incubation Time	2 h, 1 d, 2 d, 3 d, 5 d and 7 day
	Results	During the MFI increased phase (measured at 2 h and 1 d), the MFI values of the low phosphorylation level group (protein kinase inhibited group) were significantly higher (p < 0.05) than those of the control group, while the MFI values of the high phosphorylation level group (phosphatase inhibited group) were significantly lower (p < 0.05) than those of the control group.

	Source	Food Chem (2017). Figure 2. Staurosporine (Abmole Bioscience,Houston, TX, USA)
	Method	Global phosphorylation level of myofibrillar proteins
	Cell Lines	Muscles
	Concentrations	0.144 μmol/5 g
	Incubation Time	2 h, 1 d, 2 d, 3 d, 5 d and 7 day
	Results	Inhibition of protein kinase by M2066 significantly reduced the phosphorylation level of myofibrillar proteins in muscle compared to control.

	Source	Food Chemistry (2016).Figure 4. kinase inhibitors (M2066, Abmole, Houston, TX, USA)
	Method	Lactic acid concentrations
	Cell Lines
	Concentrations	0.144 μmole per 5 g muscle pieces
	Incubation Time	2h, 24h, 48h, 72h, 120h and 168h
	Results	No difference was observed in lactic acid content (Fig. 4) between C1 and C2 control groups. In addition, no difference in lactic acid was detected between phosphatase inhibition group and the two control groups at most time points, but the kinase inhibition group had lower lactic acid content than the other three groups (P < 0.05) throughout the entire experimental period.

	Source	Food Chemistry (2016).Figure 3. kinase inhibitors (M2066, Abmole, Houston, TX, USA)
	Method	PH Value
	Cell Lines
	Concentrations	0.144 μmole per 5 g muscle pieces
	Incubation Time	2h, 24h, 48h, 72h, 120h and 168h
	Results	There was no difference in pH values (Fig. 3) between the phosphatase inhibition group and the two control groups. the pH of the kinase inhibition group declined much slower during the early postmortem stage (P < 0.05) as compared to the other three groups.

	Source	Food Chemistry (2016).Figure 2. kinase inhibitors (M2066, Abmole, Houston, TX, USA)
	Method	protein phosphrylation assay
	Cell Lines
	Concentrations	0.144 μmole per 5 g muscle pieces
	Incubation Time	2h, 24h, 48h, 72h, 120h and 168h
	Results	The R630/580 values (Fig. 2) were not different between C1 and C2 control groups at any time. However, the R630/580 values of the phosphatase inhibition group were lower than those of all other groups (P < 0.05) except at 2 h. Kinase inhibition group demonstrated greater (P < 0.05) R630/580 values than those of the rest groups except at 2 and 168 h.

	Source	Food Chemistry (2016).Figure 1. kinase inhibitors (M2066, Abmole, Houston, TX, USA)
	Method	Gel electrophoresis and image analyses
	Cell Lines
	Concentrations	0.144 μmole per 5 g muscle pieces
	Incubation Time	2h, 24h, 48h, 3d, 5d and 7d
	Results	"As expected, the overall phosphorylation of sarcoplasmic proteins in the minced meat with phosphatase inhibitor was significantly higher (P < 0.05) than in control and kinase inhibitor added samples."

Protocol (for reference only)

Cell Experiment
Cell lines	PC12
Preparation method	Cells are exposed to Staurosporine for ~32 hours. Cells are fixed in 4% paraformaldehyde and stained with the DNA-binding dye Hoechst 33342. Cells are visualized under epifluorescence illumination, and the percentage of apoptotic cells (cells with condensed and fragmented DNA) is determined.
Concentrations	Dissolved in DMSO, final concentration 1 μM
Incubation time	~32 hours

Animal Experiment
Animal models	Male Mongolian gerbils or male Wistar rats subjected to transient ischemia
Formulation	Dissolved in DMSO, and diluted in saline
Dosages	~10 ng
Administration	Stereotaxically administered into the bilateral CAl subfield of the hippocampus

Chemical Information

Molecular Weight	466.53
Formula	C28H26N4O3
CAS Number	62996-74-1
Solubility (25°C)	DMSO ≥ 50 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Manns J, et al. FASEB J. Triggering of a novel intrinsic apoptosis pathway by the kinase inhibitor staurosporine: activation of caspase-9 in the absence of Apaf-1.

[2] Antonsson A, et al. Anticancer Res. Induction of apoptosis by staurosporine involves the inhibition of expression of the major cell cycle proteins at the G(2)/m checkpoint accompanied by alterations in Erk and Akt kinase activities.

[3] Zhang XD, et al. Mol Cancer Ther. Staurosporine induces apoptosis of melanoma by both caspase-dependent and -independent apoptotic pathways.

[4] Tamaoki T, et al. Biochem Biophys Res Commun. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase.

[5] Hara H, et al. J Cereb Blood Flow Metab. Staurosporine, a novel protein kinase C inhibitor, prevents postischemic neuronal damage in the gerbil and rat.