Store at -20°C. Keep tightly closed. Store in a cool dry place.
注释
Documents
Figures
Reference
J. Ma et al., Br. J. Pharmacol., 124, 756 (1998) W. Liu et al., NeuroReport 9, 2609 (1998) E. Weimann et al., Leukemia Res., 23, 751 (1999) A. Kondratyev and K. Gale, Mol. Brain Res., 75, 216 (2000)
FMK-MEA is a potent and selective p90 Ribosomal S6 Kinase (RSK) inhibitor.
IC50 & Target
p90 RSK[1]
体外研究 (In Vitro)
FMK-MEA is a water-soluble derivative of fmk. FMK-MEA treatment inhibits RSK2 kinase activity in diverse, highly invasive human cancer cell lines including 212LN, M4e, A549, and SKBR3 cells. Treatment with the RSK-specific inhibitor FMK-MEA significantly attenuates RSK2 activity, as assessed by the phosphorylation levels of Ser-386 and the consequent invasive ability of A549 cells[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.
体内研究 (In Vivo)
FMK-MEA treatment (80 mg/kg/day for 16 days by intraperitoneal injection) in highly metastatic M4e cell xenograft nude mice results in a significant attenuation of LN metastasis. FMK-MEA treatment has no effect on the tumor size, and the proliferation rate of the primary tumor[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.
分子量
399.46
Formula
C21H26FN5O2
CAS 号
1414811-15-6
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
Please store the product under the recommended conditions in the Certificate of Analysis.
参考文献
[1]. Li D, et al. The prometastatic ribosomal S6 kinase 2-cAMP response element-binding protein (RSK2-CREB) signaling pathway up-regulates the actin-binding protein fascin-1 to promote tumor metastasis. J Biol Chem. 2013 Nov 8;288(45):32528-38.
Animal Administration [1]
Mice[1]
For FMK-MEA treatment, each of the nude mice (athymic nu/nu, female, 4-6 weeks old) are injected with 0.5×106cells/100 μL of PBS submandibular to the mylohyoid muscle. On day 5 after injection, mice are divided into two groups with similar average weights with each group receiving either FMK-MEA or PBS. Each mouse is administered 80 mg/kg of FMK-MEA daily by intraperitoneal injection from 5 days after the xenograft for 16 days total. The control group receives PBS alone on the same schedule. Tumor growth is recorded. Mice are sacrificed after 16 days post drug treatment[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.
参考文献
[1]. Li D, et al. The prometastatic ribosomal S6 kinase 2-cAMP response element-binding protein (RSK2-CREB) signaling pathway up-regulates the actin-binding protein fascin-1 to promote tumor metastasis. J Biol Chem. 2013 Nov 8;288(45):32528-38.
Z-VAD-FMK (Z-VAD(OH)-FMK) 是一种 pan caspase 抑制剂。Z-VAD-FMK 不抑制泛素 C 末端水解酶 L1 (UCHL1) 活性，即使浓度高达 440 μM。
Z-VAD-FMK Chemical Structure
CAS No. : 161401-82-7
规格
价格
是否有货
数量
10 mM * 1 mL in DMSO
￥3090
In-stock
1 mg
￥875
In-stock
5 mg
￥3085
In-stock
10 mg
￥5270
In-stock
50 mg
询价
100 mg
询价
* Please select Quantity before adding items.
Z-VAD-FMK 相关产品
•相关化合物库:
Covalent Screening Library Plus
Bioactive Compound Library Plus
Apoptosis Compound Library
Anti-Cancer Compound Library
Covalent Screening Library
生物活性
Z-VAD-FMK (Z-VAD(OH)-FMK) is a well-know pan caspase inhibitor, which does not inhibit ubiquitin carboxy-terminal hydrolase L1 (UCHL1) activity even at concentrations as high as 440 μM[1].
IC50 & Target[1]
Caspase
体外研究 (In Vitro)
Z-VAD-FMK (40 μM) reverses the apoptotic effect exerted by total saponin of Solanum lyratum Thunb (TSSLT) in Hela cells. HeLa cells are pretreated with Z-VAD-FMK (40 μM) for 30 min and exposed to TSSLT (6 μg/mL) for 48 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.
Cell Viability Assay[2]
Cell Line:
HeLa cells
Concentration:
40 μM
Incubation Time:
Prtreated for 30 minutes
Result:
Prevented TSSLT-induced cell death. More than 80% cell survival was observed.
分子量
453.46
Formula
C21H28FN3O7
CAS 号
161401-82-7
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
4°C, sealed storage, away from moisture and light
*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)
溶解性数据
In Vitro:
DMSO : 100 mg/mL (220.53 mM; Need ultrasonic)
配制储备液
浓度溶剂体积质量
1 mg
5 mg
10 mg
1 mM
2.2053 mL
11.0263 mL
22.0527 mL
5 mM
0.4411 mL
2.2053 mL
4.4105 mL
10 mM
0.2205 mL
1.1026 mL
2.2053 mL
*
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 (sealed storage, away from moisture and light)。-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：
[1]. Davies CW, et al. The co-crystal structure of ubiquitin carboxy-terminal hydrolase L1 (UCHL1) with a tripeptide fluoromethyl ketone (Z-VAE(OMe)-FMK). Bioorg Med Chem Lett. 2012 Jun 15;22(12):3900-4.
[2]. Liu HR, et al. Antiproliferative activity of the total saponin of Solanum lyratum Thunb in Hela cells by inducing apoptosis. Pharmazie. 2008 Nov;63(11):836-42.
Z-VAD(OMe)-FMK (Z-Val-Ala-Asp(OMe)-FMK) is a cell-permeable and irreversible pan-caspase inhibitor[1]. Z-VAD(OMe)-FMK is an ubiquitin carboxy-terminal hydrolase L1 (UCHL1) inhibitor. Z-VAD(OMe)-FMK irreversibly modifies UCHL1 by targeting the active site of UCHL1[2].
IC50 & Target[1]
Caspase
体外研究 (In Vitro)
Z-VAD(OMe)-FMK (Z-Val-Ala-Asp(OMe)-FMK) is a broad-spectrum caspase inhibitor, has been shown to inhibit the intracellular activation of caspase-like proteases. The injection of Z-VAD(OMe)-FMK suppresses the caspase-3 activity in lung tissues, and significantly decreases the number of terminal dUTP nick-end labeling-positive cells[1]. Z-VAD(OMe)-FMK effectively inhibits UCHL1’s reaction with hemagglutinin-tagged ubiquitin vinylmethyl ester (HA-UbVME) at the concentration of 100 μM[2]. Z-VAD(OMe)-FMK is administered intraperitoneally at 1 hour before and 6 hours after SAH. Expression of caspase-3 and positive TUNEL is examined as markers for apoptosis. Z-VAD(OMe)-FMK suppresses TUNEL and caspase-3 staining in endothelial cells, decreases caspase-3 activation, reduces BBB permeability, relieves vasospasm, abolishes brain edema, and improves neurological outcome[3]. Z-VAD(OMe)-FMK is a cell-permeable caspase inhibitor, efficiently blocks cell death induced by SMN deficiency[4].
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.
体内研究 (In Vivo)
The survival rate of mice is prolonged significantly by the injection of Z-VAD(OMe)-FMK (Z-Val-Ala-Asp(OMe)-FMK). All mice succumbed to LPS within 30 hours. By contrast, the mice treated with Z-VAD(OMe)-FMK survive significantly longer and 27% of the mice survived more than 7 days[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.
分子量
467.49
Formula
C22H30FN3O7
CAS 号
187389-52-2
Sequence
Z-Val-Ala-Asp(OMe)-FMK
Sequence Shortening
ZVA-D(OMe)-FMK
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Kawasaki M, et al. Protection from lethal apoptosis in lipopolysaccharide-induced acute lung injury in mice by a caspaseinhibitor. Am J Pathol. 2000 Aug;157(2):597-603.
[2]. Davies CW, et al. The co-crystal structure of ubiquitin carboxy-terminal hydrolase L1 (UCHL1) with a tripeptide fluoromethyl ketone (Z-VAE(OMe)-FMK). Bioorg Med Chem Lett. 2012 Jun 15;22(12):3900-4.
[3]. Park S, et al. Neurovascular protection reduces early brain injury after subarachnoid hemorrhage. Stroke. 2004 Oct;35(10):2412-7.
[4]. Ilangovan R, et al. Inhibition of apoptosis by Z-VAD-fmk in SMN-depleted S2 cells. J Biol Chem. 2003 Aug 15;278(33):30993-9.
Cell Assay [4]
PCR products containing coding sequences for the dSMN (forward primer: 5′-TAA TAC GAC TCA CTA TAG GG AAG ACG TAC GAC GAG TCG-3′; and reverse primer: 5′-TAA TAC GAC TCA CTA TAG GG GTG GTG CTG GCT TCT TTC-3′; product length, 601bps; bold and italics letters represent T7 promoter sequences) and control Drosophila Presenilin (dPsn) gene (forward primer: 5′-TAA TAC GAC TCA CTA TAG GG TG GCT GCT GTC AAT CTC-3′; and reverse primer: 5′-TAA TAC GAC TCA CTA TAG GG CGA TAG CAA CGC TTC TTG-3′; product length: 543bps) are obtained and gel-purified. Double-stranded RNAs (dsRNA) are generated by transcription with Ribomax T7 Transcription kit and digested with Rnase-free DNase. The dsRNA products are ethanol precipitated and annealed by incubation at 65°C for 30 min and then slowly allowed to cool at room temperature. The annealed dsRNA products are analyzed on a 1% agaorse gel to ensure the majority of dsRNA existed as a single band. The dsRNA (2 μg) and/or plasmid DNAs (2 μg) are introduced into cells by using Cellfectin. Caspase inhibition is achieved by using 50 μM of Z-VAD(OMe)-FMK in the culture medium[4].
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.
Animal Administration [1][3]
Mice[1] Mice used in this study are 5- to 6-week-old (20 to 22 g) ICR males. Mice are injected with 30 mg/kg LPS from E. coli serotype O111:B4 through the tail vein. A single intravenous injection of Z-VAD(OMe)-FMK (0.25 mg) is made 15 minutes before LPS injection, followed by three intravenous injections of Z-VAD(OMe)-FMK (0.1 mg each) per hour. Control mice are injected with the same volume of 1% DMSO in sterile saline. Rats[3] Male Sprague-Dawley rats weighing 300 to 350 g are anesthetized with α-chloralose (40 mg/kg IP) and urethane (400 mg/kg IP). Animals are intubated, and respiration is maintained with a small animal respirator. Rectal temperature is maintained at 37±0.5°C with a heating pad. The left external carotid artery is isolated and a 4.0 monofilament nylon suture is inserted through the internal carotid artery to perforate the middle cerebral artery. SAH is confirmed at autopsy in each rat. Sham-operated rats underwent the same procedures except that the suture is withdrawn after resistance is felt. Z-VAD(OMe)-FMK (50 μM per 0.3 mL) is injected intraperitoneally at 1 hour before and 6 hours after SAH induction. In vehicle group, rats underwent SAH induction and are treated with the same volume of vehicle (DMSO diluted in physiological buffer solution). No treatment is applied in sham-operated animals.
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.
参考文献
[1]. Kawasaki M, et al. Protection from lethal apoptosis in lipopolysaccharide-induced acute lung injury in mice by a caspaseinhibitor. Am J Pathol. 2000 Aug;157(2):597-603.
[2]. Davies CW, et al. The co-crystal structure of ubiquitin carboxy-terminal hydrolase L1 (UCHL1) with a tripeptide fluoromethyl ketone (Z-VAE(OMe)-FMK). Bioorg Med Chem Lett. 2012 Jun 15;22(12):3900-4.
[3]. Park S, et al. Neurovascular protection reduces early brain injury after subarachnoid hemorrhage. Stroke. 2004 Oct;35(10):2412-7.
[4]. Ilangovan R, et al. Inhibition of apoptosis by Z-VAD-fmk in SMN-depleted S2 cells. J Biol Chem. 2003 Aug 15;278(33):30993-9.
Boc-D-FMK is a cell-permeable, irreversible and broad spectrum caspase inhibitor. Boc-D-FMK inhibits apoptosis stimulated by TNF-α with an IC50 of 39 µM.
IC50 & Target[1]
Caspase
体外研究 (In Vitro)
Apoptosis is a pathway of cell death orchestrated by a family of proteases called caspases. Boc-D-fmk inhibits TNFα-stimulated reactive oxygen species (ROS) generation. Boc-D-FMK inhibits apoptosis stimulated by TNF-α with an IC50 of 39 µM[1]. BocD-fmk at 50 µM prevents genistein-induced apoptosis of p815 cells. Confocal microscopy shows that the release of mitochondrial apoptotic factors is inhibited by BocD-fmk[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.
体内研究 (In Vivo)
Boc-D-FMK-fmk effectively attenuates the hepatocyte apoptosis in bile duct-ligated rats and may improve the survival rates after endotoxin challenge[3]. A single injection of Boc-D-FMK results in longterm protection of MNs against root avulsion-induced death for more than 8 weeks and the Boc-D-FMK-treated MNs are able to regenerate their axons into an implanted PN graft and reinnervate the target muscle[4].
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.
分子量
263.26
Formula
C11H18FNO5
CAS 号
634911-80-1
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Cowburn AS, et al. z-VAD-fmk augmentation of TNF alpha-stimulated neutrophil apoptosis is compound specific and does not involve the generation of reactive oxygen species.
[2]. Yee SB, et al. zVAD-fmk, unlike BocD-fmk, does not inhibit caspase-6 acting on 14-3-3/Bad pathway in apoptosis of p815 mastocytoma cells. Exp Mol Med. 2006 Dec 31;38(6):634-42.
[3]. Sheen-Chen SM, et al. Effect of Boc-D-Fmk on hepatocyte apoptosis after bile duct ligation in rat and survival rate after endotoxin challenge. J Gastroenterol Hepatol. 2008 Aug;23(8 Pt 1):1276-9.
[4]. Chan YM, et al. Inhibition of caspases promotes long-term survival and reinnervation by axotomized spinal motoneurons of denervated muscle in newborn rats. Exp Neurol. 2003 Jun;181(2):190-203.
Animal Administration [3]
Rats: Boc-D-FMK is dissolved in DMSO. Male Sprague-Dawley rats group 1 (OBBOC-D) undergo common bile duct ligation and simultaneously treatment with Boc-D-FMK. The first dose of Boc-D-FMK (1.5 mg/kg) is injected into the inferior vena cava immediately after bile duct ligation. Subsequent doses of Boc-DFMK (1.5 mg/kg twice daily) are given intraperitoneally on the first and second postoperative days. The last dose (1.5 mg/kg) is given on the morning of the third postoperative day[3].
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.
参考文献
[1]. Cowburn AS, et al. z-VAD-fmk augmentation of TNF alpha-stimulated neutrophil apoptosis is compound specific and does not involve the generation of reactive oxygen species.
[2]. Yee SB, et al. zVAD-fmk, unlike BocD-fmk, does not inhibit caspase-6 acting on 14-3-3/Bad pathway in apoptosis of p815 mastocytoma cells. Exp Mol Med. 2006 Dec 31;38(6):634-42.
[3]. Sheen-Chen SM, et al. Effect of Boc-D-Fmk on hepatocyte apoptosis after bile duct ligation in rat and survival rate after endotoxin challenge. J Gastroenterol Hepatol. 2008 Aug;23(8 Pt 1):1276-9.
[4]. Chan YM, et al. Inhibition of caspases promotes long-term survival and reinnervation by axotomized spinal motoneurons of denervated muscle in newborn rats. Exp Neurol. 2003 Jun;181(2):190-203.
Z-DEVD-FMK is a specific and irreversible caspase-3 inhibitor with an IC50 of 18 μM[1].
IC50 & Target[1]
Caspase-3
18 μM (IC50)
体外研究 (In Vitro)
N27 cells are exposed to MPP+ in the absence or presence of 50 μM Z-DIPD-FMK or 100 μM Z-DEVD-FMK or 50 μM Z-LEHD-FMK and then caspase-9 and caspase-3 enzymatic activities are determined by enzymatic assay at 12 and 24 h following exposure, respectively. Exposure to 300 μM MPP+ for 24 h in N27 cells results in an approximately 2.5-fold increase in caspase-3 enzyme activity. MPP+-induced increases in caspase-3 enzyme activity are significantly blocked by 50 μM Z-DIPD-FMK, 100 μM Z-DEVD-FMK, and 50 μM Z-LEHD-FMK[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.
体内研究 (In Vivo)
Early Z-DEVD-FMK (160 ng) treatment improves motor and cognitive function after traumatic CNS injury induced by severe controlled cortical impact (CCI) in the mouse[2]. Treatment with Z-DEVD-FMK (160 ng) significantly improves neurological outcome when compared with traumatized animals treated with DMSO vehicle (p<0.01)[3].
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.
分子量
668.66
Formula
C30H41FN4O12
CAS 号
210344-95-9
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Kanthasamy AG, et al. A novel peptide inhibitor targeted to caspase-3 cleavage site of a proapoptotic kinase protein kinase C delta (PKCdelta) protects against dopaminergic neuronal degeneration in Parkinson’s disease models. Free Radic Biol Med. 2006 Nov 15;41(10):1578-89.
[2]. Knoblach SM, et al. Caspase inhibitor z-DEVD-fmk attenuates calpain and necrotic cell death in vitro and after traumatic brain injury. J Cereb Blood Flow Metab. 2004 Oct;24(10):1119-32.
[3]. Yakovlev AG, et al. Activation of CPP32-like caspases contributes to neuronal apoptosis and neurological dysfunction after traumatic brain injury. J Neurosci. 1997, 17(19), 7415-7424.
[4]. Huang MY, et al. Chemotherapeutic agent CPT-11 eliminates peritoneal resident macrophages by inducing apoptosis. Apoptosis. 2016 Feb;21(2):130-42.
Cell Assay [1]
N27 cells and primary mesencephalic neurons are exposed to either 10-100 μM 6-OHDA or 10-300 μM MPP+ in the presence or absence of 0.1-50 μM Z-DIPD-FMK or 0.1-100 μM Z-DEVD-FMK or 50 μM Z-IETD-FMK or Z-LEHD-FMK for the duration of the experiment. N27 cells are incubated with 100 μM 6-OHDA for 24 h or 300 μM MPP+ for 36 h in the presence or absence of 50 μM Z-DEVD-FMK and cell death is determined by MTT assay, which is widely used to assess cell viability. After treatment, the cells are incubated in serum-free medium containing 0.25 mg/mL MTT for 3 h at 37°C. Formation of formazan from tetrazolium is measured at 570 nm with a reference wavelength at 630 nm using a SpectraMax microplate reader[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.
Animal Administration [2][3]
Mice[2] Male C57Bl/6 mice (20-25 g) are used. For treatment with Z-DEVD-fmk or vehicle after CCI, mice are placed in a stereotaxic apparatus, and the CCI wound is reopened for intracerebroventricular injection. Either Z-DEVD-FMK (160 ng in 2 μL DMSO), or DMSO vehicle is injected over a 5-minute period. Rats[3] Male Sprague Dawley rats (425±25 g) are used. DMSO (5 μL) vehicle or Z-DEVD-FMK (160 ng in 5 μL of DMSO) is administered at a controlled rate of 0.5 μL/min via an infusion pump at 30 min before and at 6 and 24 hr after TBI. At the designated time periods after injury, animals are decapitated under NSC 10816 anesthesia (100 mg/kg, i.p.), and the brains are removed rapidly and dissected. Sham-operated (control) animals received anesthesia and surgery but are not subjected to trauma. Tissue samples are collected 1, 4, 12, 24, and 72 hr after TBI. Samples are frozen on dry ice and kept at −85°C.
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.
参考文献
[1]. Kanthasamy AG, et al. A novel peptide inhibitor targeted to caspase-3 cleavage site of a proapoptotic kinase protein kinase C delta (PKCdelta) protects against dopaminergic neuronal degeneration in Parkinson’s disease models. Free Radic Biol Med. 2006 Nov 15;41(10):1578-89.
[2]. Knoblach SM, et al. Caspase inhibitor z-DEVD-fmk attenuates calpain and necrotic cell death in vitro and after traumatic brain injury. J Cereb Blood Flow Metab. 2004 Oct;24(10):1119-32.
[3]. Yakovlev AG, et al. Activation of CPP32-like caspases contributes to neuronal apoptosis and neurological dysfunction after traumatic brain injury. J Neurosci. 1997, 17(19), 7415-7424.
[4]. Huang MY, et al. Chemotherapeutic agent CPT-11 eliminates peritoneal resident macrophages by inducing apoptosis. Apoptosis. 2016 Feb;21(2):130-42.
Z-IETD-FMK (Z-IE(OMe)TD(OMe)-FMK) 是一种具有细胞渗透作用的选择性 caspase-8 抑制剂。Z-IETD-FMK 也是颗粒酶 B (granzyme B) 抑制剂。
Z-IETD-FMK Chemical Structure
CAS No. : 210344-98-2
规格
价格
是否有货
数量
10 mM * 1 mL in DMSO
￥4321
In-stock
1 mg
￥1400
In-stock
5 mg
￥3000
In-stock
10 mg
询价
50 mg
询价
* Please select Quantity before adding items.
Z-IETD-FMK 相关产品
•相关化合物库:
Bioactive Compound Library Plus
Peptidomimetic Library
Peptide Library
生物活性
Z-IETD-FMK (Z-IE(OMe)TD(OMe)-FMK) is a selective and cell permeable caspase-8 inhibitor[1]. Z-IETD-FMK is also a granzyme B inhibitor[5].
IC50 & Target[1]
Caspase-8
体外研究 (In Vitro)
Z-IETD-FMK causes full inhibition only of the proapoptotic effect of TNFα with an IC50 of 0.46 μM[1]. Z-IETD-FMK and Z-VAD-FMK at non-toxic doses are found to be immunosuppressive and inhibit human T cell proliferation induced by mitogens and IL-2. They are shown to block NF-κB in activated primary T cells, but have little inhibitory effect on the secretion of IL-2 and IFN-γ during T cell activation[2]. Z-IETD-FMK inhibits the cleavage of caspase-8 and only partially inhibits the cleavage of caspase-3 and PARP. Z-IETD-FMK can prevent the execution of apoptosis in retinal cells exposed to different apoptotic stimuli[3].
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.
体内研究 (In Vivo)
Pharmacological inhibition of caspase-8 by z-IETD-FMK robustly reduces tumour outgrowth and this is closely associated with a reduction in the release of pro-inflammatory cytokines, IL-6, TNF-α, IL-18, IL-1α, IL-33, but not IL-1β. Furthermore, inhibition of caspase-8 reduces the recruitment of innate suppressive cells, such as myeloid-derived suppressor cells, but not of regulatory T cells to lungs of tumour-bearing mice[4].
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.
分子量
654.68
Formula
C30H43FN4O11
CAS 号
210344-98-2
Sequence
Z-Ile-Glu-Thr-Asp-FMK
Sequence Shortening
ZIETDFMK
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Cowburn AS, et al. z-VAD-fmk augmentation of TNF alpha-stimulated neutrophil apoptosis is compound specific and does not involve the generation of reactive oxygen species.
[2]. Lawrence CP, et al. Suppression of human T cell proliferation by the caspase inhibitors, z-VAD-FMK and z-IETD-FMK is independent of their caspase inhibition properties. Toxicol Appl Pharmacol. 2012 Nov 15;265(1):103-12.
[3]. Tezel G, et al. Inhibition of caspase activity in retinal cell apoptosis induced by various stimuli in vitro. Invest Ophthalmol Vis Sci. 1999 Oct;40(11):2660-7.
[4]. Terlizzi M, et al. Pharmacological inhibition of caspase-8 limits lung tumour outgrowth. Br J Pharmacol. 2015 Aug;172(15):3917-28.
[5]. Yang J, et al. Granzyme B Is an Essential Mediator in CD8+ T Cell Killing of Theileria parva-Infected Cells.Infect Immun. 2018 Dec 19;87(1). pii: e00386-18.
Cell Assay [2]
T cell proliferation following mitogen stimulation is determined using [3H]-thymidine incorporation. In brief, PBMCs or purified T cells are seeded at 1×106 cells/mL in 96 well plates and stimulated with either PHA (5 μg/mL or co-stimulated with anti-CD3 mAb (5 μg/mL) and anti-CD28 mAb (2.5 μg/mL) in the presence or absence of caspase inhibitor Z-IETD-FMK. The cells are cultured for 72 h with the last 16 h pulsed with [[3H]-labelled methyl-thymidine (0.037 MBq) prior to harvest onto glass fibre filter mats using a Tomtec automated multi-well harvester[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.
Animal Administration [4]
Mice: Mice are divided into three groups: (1) naive, non-treated, mice; (2) CTR (control), i.t. instilled with NMU; and (3) lung cancer-bearing mice treated with Z-IETD-FMK (0.5 μg per mouse). The involvement of caspase-8 in lung cancer development is the determined at different time points (3, 7 and 28 days)[4].
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.
参考文献
[1]. Cowburn AS, et al. z-VAD-fmk augmentation of TNF alpha-stimulated neutrophil apoptosis is compound specific and does not involve the generation of reactive oxygen species.
[2]. Lawrence CP, et al. Suppression of human T cell proliferation by the caspase inhibitors, z-VAD-FMK and z-IETD-FMK is independent of their caspase inhibition properties. Toxicol Appl Pharmacol. 2012 Nov 15;265(1):103-12.
[3]. Tezel G, et al. Inhibition of caspase activity in retinal cell apoptosis induced by various stimuli in vitro. Invest Ophthalmol Vis Sci. 1999 Oct;40(11):2660-7.
[4]. Terlizzi M, et al. Pharmacological inhibition of caspase-8 limits lung tumour outgrowth. Br J Pharmacol. 2015 Aug;172(15):3917-28.
[5]. Yang J, et al. Granzyme B Is an Essential Mediator in CD8+ T Cell Killing of Theileria parva-Infected Cells.Infect Immun. 2018 Dec 19;87(1). pii: e00386-18.
FMK is a an irreversible RSK2 kinase inhibitor, that covalently modifies the C-terminal kinase domain of RSK.
体外研究 (In Vitro)
Pretreatment of ARVMs with 3 μM fmk attenuates the increase in Ser386 phosphorylation, but it has no inhibitory effect on the increase in Thr577 phosphorylation[1]. FMK inhibits relatively few protein kinases in the panel, although it does inhibit protein tyrosine kinases, such as Src, Lck, Yes and Eph-A2, as well as S6K1. FMK will not inhibit RSK if the N-terminal kinase domain are activated by a mechanism that is independent of the C-terminal domain[2]. Fmk potently inactivates the CTD auto-kinase activity of RSK1 and RSK2 with high specificity in mammalian cells. Targeting RSK2 by a specific small molecule RSK inhibitor fmk attenuates FGFR3-induced cytokine-independent growth in Ba/F3 cells. FMK inhibits cytokine-independent proliferation of Ba/F3 cells conferred by FGFR3[3].
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.
分子量
342.37
Formula
C18H19FN4O2
CAS 号
821794-92-7
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Cuello F, et al. Evidence for direct regulation of myocardial Na+/H+ exchanger isoform 1 phosphorylation and activity by 90-kDa ribosomal S6 kinase (RSK): effects of the novel and specific RSK inhibitor fmk on responses to alpha1-adrenergic stimulation.
[2]. Bain J, et al. The selectivity of protein kinase inhibitors: a further update. Biochem J. 2007 Dec 15;408(3):297-315.
[3]. Kang S, et al. FGFR3 activates RSK2 to mediate hematopoietic transformation through tyrosine phosphorylation of RSK2 and activation of the MEK/ERK pathway. Cancer Cell. 2007 Sep;12(3):187-9.
Kinase Assay [3]
The S6 peptide kinase assay is carried out according to the manufacturer’s protocol using RSK2 immunoprecipitates. To determine the ability of FGFR3 to phosphorylate RSK2, 500 ng of purified recombinant RSK2 variants are incubated with 500 ng of recombinant active FGFR3 in 10 mM HEPES (pH 7.5), 150 mM NaCl, 1 mM DTT, 0.01% Trixton-X-100, 10 mM MnCl2, and 200 μM ATP for 30 min at 30°C. Phosphorylation of Y529 RSK2 is detected by specific phospho-antibody. To determine kinase activity of RSK2 CTD variants, purified recombinant RSK2 CTD proteins (500 nM) are incubated with 500 nM of active ERK in 20 mM HEPES [pH 8.0], 10 mM MgCl2, 2 mM tris-(2-carboxyethyl)-phosphine (TCEP), and 200 μM ATP for 1 hr at 30°C. Kinase reactions are initiated by the addition of 5 μCi of [γ-32P] ATP and 100 μM peptide substrate (CTD-tide), followed by incubation for 20 min at room temperature. Kinase activity is determined using the standard disk phospho-cellulose assay.
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.
Cell Assay [3]
RSK2 expressing Ba/F3 cell lines are generated by retroviral transduction as described by using Ba/F3 cells stably expressing FGFR3 TDII with pMSCV-puro plasmids encoding myc-tagged RSK2 variants, followed by antibiotic selection. For cell viability assays, 1×105 Ba/F3 cells stably expressing FGFR3 are cultured in 24-well plates with media containing increasing concentrations of FMK, acidic FGF (10 nM), and heparin (30 μg/mL) in the absence of IL-3. The relative cell viability at each experimental time point is determined by using the Celltiter96AQueous One solution proliferation kit.
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.
参考文献
[1]. Cuello F, et al. Evidence for direct regulation of myocardial Na+/H+ exchanger isoform 1 phosphorylation and activity by 90-kDa ribosomal S6 kinase (RSK): effects of the novel and specific RSK inhibitor fmk on responses to alpha1-adrenergic stimulation.
[2]. Bain J, et al. The selectivity of protein kinase inhibitors: a further update. Biochem J. 2007 Dec 15;408(3):297-315.
[3]. Kang S, et al. FGFR3 activates RSK2 to mediate hematopoietic transformation through tyrosine phosphorylation of RSK2 and activation of the MEK/ERK pathway. Cancer Cell. 2007 Sep;12(3):187-9.
ATG4B or autophagin-1 is a cysteine protease that cleaves ATG8 family proteins. ATG4B plays essential roles in the autophagosome formation and the autophagy pathway. FMK 9a shows strong inhibition of ATG4B, with IC50 values of 80 and 73 nM in the TR-FRET and cellular-based LRA assays, respectively. LC−MS/MS study indicates that FMK 9a forms an irreversible covalent bond with the more reactive thiol group of Cys74 located at the catalytic site of ATG4B and thus inactivates ATG4B proteolytic activity[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.
体内研究 (In Vivo)
FMK 9a shows moderate solubility (LYSA: 41 μg/mL) and high human and mouse liver microsome clearances of 13.9 and 70 mL/kg per minute[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.
分子量
392.42
Formula
C23H21FN2O3
CAS 号
1955550-51-2
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Qiu Z, et al. Discovery of Fluoromethylketone-Based Peptidomimetics as Covalent ATG4B (Autophagin-1) Inhibitors. ACS Med Chem Lett. 2016 Jun 25;7(8):802-6.
Kinase Assay [1]
FMK 9a is dissolved in DMSO. 5 μL of serial-diluted FMK 9a (1% DMSO) in assay buffer is added into 5 μL of purified ATG4B (final 0.1 nM) in 384-well plate, and the solution is incubated at room temperature (rt) for 30 min. Then, 5 μL of His-GATE-16-GST (final 20 nM) is added into the wells, and the solution is incubated for another 30 min. 5 μL of detection solution (final 2 nM of Eu-anti-His and 20 nM of Ulight-anti-GST) is added, and the resulting mixture is incubated at rt for 40 min[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.
参考文献
[1]. Qiu Z, et al. Discovery of Fluoromethylketone-Based Peptidomimetics as Covalent ATG4B (Autophagin-1) Inhibitors. ACS Med Chem Lett. 2016 Jun 25;7(8):802-6.
Z-YVAD-FMK is a cell-permeable caspase-1 and -4 inhibitor with anti-inflammatory and anti-tumor activities[1].
IC50 & Target
Caspase
体外研究 (In Vitro)
Z-YVAD-FMK (100 μM; 24 hours) significantly downregulated the growth inhibition induced by butyrate in Caco-2 cells[1]. Z-YVAD-FMK (20 μM; pre 1 hour; 24 hours) attenuates the apoptotic induction of III-10 on both HepG2 and BEL-7402 cells, the apoptotic rate of -10 on HepG2 cells is reduced by Z-VAD-FMK from 19.88% to 8.34%, while that on BEL-7402 cells is reduced from 17.56% to 11.98%[4].
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.
Cell Viability Assay[1]
Cell Line:
Caco-2 cells
Concentration:
0-100 μM
Incubation Time:
24 hours
Result:
Inhibited Caco-2 cells growth.
Apoptosis Analysis[1]
Cell Line:
BEL-7402 and HepG2 cells
Concentration:
20 μM
Incubation Time:
Pre 1 hour; 24 hours
Result:
Induced a caspase-dependent apoptosis in cells.
分子量
630.66
Formula
C31H39FN4O9
CAS 号
210344-97-1
Sequence
Z-Tyr-Val-Ala-Asp-FMK
Sequence Shortening
Z-YVAD-FMK
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Li H1,et al.Aluminum hydroxide adjuvants activate caspase-1 and induce IL-1beta and IL-18 release.J Immunol. 2007 Apr 15;178(8):5271-6.
[2]. Avivi-Green C, et al. Different molecular events account for butyrate-induced apoptosis in two human colon cancer cell lines.J Nutr. 2002 Jul;132(7):1812-8.
