Decanoyl-RVKR-CMK
(Synonyms: Furin Inhibitor I) 目录号 : GC15108
A proprotein convertase inhibitor
Cas No.:150113-99-8
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >95.00%
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- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
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Preparation Method |
PC12 cells were rinsed with serum-free medium and treated with serum-free medium (with or without 100 µm Decanoyl-RVKR-CMK) for 3 h (basal release) and finally with serum-free medium containing 50 mm KCl (with or without 100 µm Decanoyl-RVKR-CMK) for 3 h. |
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Reaction Conditions |
100 µm Decanoyl-RVKR-CMK for 3h |
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Applications |
PC12 cells were treated with the PC enzyme inhibitor Decanoyl-RVKR-CMK to partially block the regulated release of VGF. VGF treatment in cell extracts and culture medium from Decanoyl-RVKR-CMK-treated PC12 reduced the culture, indicating extensive inhibition of invertase activity at these Decanoyl-RVKR-CMK concentrations. |
| Animal experiment [2]: | |
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Animal models |
K5-PACE4 transgenic mice |
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Preparation Method |
In mice topically treated with the hyperplasiogenic phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), a 2-day topical treatment of Decanoyl-RVKR-CMK at 300 uM. |
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Dosage form |
Decanoyl-RVKR-CMK was locally treated at 300 µM for 2 days; Once daily for 3 weeks |
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Applications |
When used Decanoyl-RVKR-CMK. PACE4 activity in skin squamous cell carcinoma cell lines resulted in impaired insulin-like growth factor 1 receptor maturation, diminished its intrinsic tyrosine kinase activity, and decreased tumor cell proliferation. Two-stage skin chemical carcinogenesis experiments, together with topical applications of CMK, demonstrated that Decanoyl- Decanoyl-RVKR-CMK markedly reduced tumor incidence, tumor multiplicity, and metastasis, pointing to a significant delay in tumor progression in wild-type and PACE4 transgenic mice. |
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References: [1]: Garcia AL, Han SK, et,al. A prohormone convertase cleavage site within a predicted alpha-helix mediates sorting of the neuronal and endocrine polypeptide VGF into the regulated secretory pathway. J Biol Chem. 2005 Dec 16;280(50):41595-608. doi: 10.1074/jbc.M509122200. Epub 2005 Oct 12. PMID: 16221685. | |
Decanoyl-RVKR-CMK is a subtilisin/Kex2p-like proprotein convertase inhibitor; blocks activity of all seven convertases (PC1, PC2, PC4, PACE4, PC5, PC7 and furin)[4]. Decanoyl-RVKR-CMK inhibits cleavage of SARS-CoV-2 spike protein by furin and blocks viral cell entry (IC50 ? = 57 nM in plaque reduction assay)[9].
PC12 cells were treated with the PC enzyme inhibitor Decanoyl-RVKR-CMK to partially block the regulated release of VGF. VGF treatment in cell extracts and culture medium from Decanoyl-RVKR-CMK-treated PC12 reduced the culture, indicating extensive inhibition of invertase activity at these Decanoyl-RVKR-CMK concentrations[2]. Decanoyl-RVKR-CMK inhibits cleavage of glycoprotein B of human cytomegalovirus[8]. Decanoyl-RVKR-CMK promotes ciliated cell differentiation and has no effect on the ciliary beat frequency in air-liquid interface (ALI) cultures of human nasal epithelial cells (HNECs). CMK considerably increases ciliogenesis-related gene expression. CMK inhibited Notch1 processing and promoted regeneration and ciliogenesis of the mouse nasal respiratory epithelium after ZnSO4 injury[7]. In LoVo cell, Decanoyl-RVKR-CMK strongly reduced the recovery of sAPPα, Overexpression of Decanoyl-RVKR-CMK at high concentrations did not completely eliminate sAPP α-secretion[3]. Decanoyl-RVKR-CMK blocked entry of MERS-CoV harboring an S protein lacking furin cleavage sites; it even blocked entry into furin-deficient LoVo cells. In addition, Decanoyl-RVKR-CMK inhibited not only the enzymatic activity of furin but also those of cathepsin L, cathepsin B, trypsin, papain, and TMPRSS2[5]. Decanoyl-RVKR-CMK inhibits HIV-2ROD replication by blocking envelope glycoprotein precursor processing in the Jurkat lymphocyte cell[1].
When used Decanoyl-RVKR-CMK. PACE4 activity in skin squamous cell carcinoma cell lines resulted in impaired insulin-like growth factor 1 receptor maturation, diminished its intrinsic tyrosine kinase activity, and decreased tumor cell proliferation. Two-stage skin chemical carcinogenesis experiments, together with topical applications of CMK, demonstrated that Decanoyl-RVKR-CMK markedly reduced tumor incidence, tumor multiplicity, and metastasis, pointing to a significant delay in tumor progression in wild-type and PACE4 transgenic mice[6].
References:
[1]:Bahbouhi B, Bendjennat M, et,al. Inhibition of HIV-2(ROD) replication in a lymphoblastoid cell line by the alpha1-antitrypsin Portland variant (alpha1-PDX) and the decRVKRcmk peptide: comparison with HIV-1(LAI). Microbes Infect. 2001 Nov;3(13):1073-84. doi: 10.1016/s1286-4579(01)01467-8. PMID: 11709287.
[2]: Garcia AL, Han SK, et,al. A prohormone convertase cleavage site within a predicted alpha-helix mediates sorting of the neuronal and endocrine polypeptide VGF into the regulated secretory pathway. J Biol Chem. 2005 Dec 16;280(50):41595-608. doi: 10.1074/jbc.M509122200. Epub 2005 Oct 12. PMID: 16221685.
[3]: Lopez-Perez E, Zhang Y, et,al. Constitutive alpha-secretase cleavage of the beta-amyloid precursor protein in the furin-deficient LoVo cell line: involvement of the pro-hormone convertase 7 and the disintegrin metalloprotease ADAM10. J Neurochem. 2001 Mar;76(5):1532-9. doi: 10.1046/j.1471-4159.2001.00180.x. PMID: 11238737.
[4]: Fugère M, Day R. Cutting back on pro-protein convertases: the latest approaches to pharmacological inhibition. Trends Pharmacol Sci. 2005 Jun;26(6):294-301. doi: 10.1016/j.tips.2005.04.006. PMID: 15925704; PMCID: PMC7119077.
[5]: Matsuyama S, Shirato K, et,al. Middle East Respiratory Syndrome Coronavirus Spike Protein Is Not Activated Directly by Cellular Furin during Viral Entry into Target Cells. J Virol. 2018 Sep 12;92(19):e00683-18. doi: 10.1128/JVI.00683-18. PMID: 30021905; PMCID: PMC6146822.
[6]: Bassi DE, Zhang J, et,al. Proprotein convertase inhibition results in decreased skin cell proliferation, tumorigenesis, and metastasis. Neoplasia. 2010 Jul;12(7):516-26. doi: 10.1593/neo.92030. PMID: 20651981; PMCID: PMC2907578.
[7]: Lee SN, Choi IS, Kim HJ, Yang EJ, Min HJ, Yoon JH. Proprotein convertase inhibition promotes ciliated cell differentiation - a potential mechanism for the inhibition of Notch1 signalling by decanoyl-RVKR-chloromethylketone. J Tissue Eng Regen Med. 2017 Sep;11(9):2667-2680. doi: 10.1002/term.2240. Epub 2016 Nov 22. PMID: 27878968; PMCID: PMC6214225.
[8]:Vey M, Sch?fer W, et,al. Proteolytic processing of human cytomegalovirus glycoprotein B (gpUL55) is mediated by the human endoprotease furin. Virology. 1995 Jan 10;206(1):746-9. doi: 10.1016/s0042-6822(95)80002-6. PMID: 7726996.
[9]:Cheng YW, Chao TL, et,al. Furin Inhibitors Block SARS-CoV-2 Spike Protein Cleavage to Suppress Virus Production and Cytopathic Effects. Cell Rep. 2020 Oct 13;33(2):108254. doi: 10.1016/j.celrep.2020.108254. Epub 2020 Sep 23. PMID: 33007239; PMCID: PMC7510585.
Decanoyl-RVKR-CMK 是一种枯草杆菌蛋白酶/Kex2p 样前蛋白转化酶抑制剂；阻断所有七种转化酶（PC1、PC2、PC4、PACE4、PC5、PC7 和弗林蛋白酶）的活性[4]。癸酰基-RVKR-CMK 抑制弗林蛋白酶对 SARS-CoV-2 刺突蛋白的切割并阻断病毒细胞进入（IC50 = 57 nM，在噬菌斑减少试验中）[9] .
PC12 细胞用 PC 酶抑制剂 Decanoyl-RVKR-CMK 处理，以部分阻断 VGF 的调节释放。来自癸酰基-RVKR-CMK 处理的 PC12 的细胞提取物和培养基中的 VGF 处理减少了培养物，表明在这些癸酰基-RVKR-CMK 浓度下对转化酶活性的广泛抑制[2]。 Decanoyl-RVKR-CMK 抑制人巨细胞病毒糖蛋白 B 的裂解[8]。 Decanoyl-RVKR-CMK 促进纤毛细胞分化，并且对人鼻上皮细胞 (HNEC) 的气液界面 (ALI) 培养物中的纤毛搏动频率没有影响。 CMK 显着增加与纤毛发生相关的基因表达。 CMK 抑制 Notch1 加工，促进 ZnSO4 损伤后小鼠鼻呼吸道上皮的再生和纤毛生成[7]。在 LoVo 细胞中，Decanoyl-RVKR-CMK 强烈降低了 sAPPα 的恢复，高浓度 Decanoyl-RVKR-CMK 的过表达并没有完全消除 sAPP α-分泌[3]。癸酰基-RVKR-CMK 阻断含有缺乏弗林蛋白酶切割位点的 S 蛋白的 MERS-CoV 的进入；它甚至阻止进入缺乏弗林蛋白酶的 LoVo 细胞。此外，Decanoyl-RVKR-CMK 不仅抑制弗林蛋白酶的酶活性，还抑制组织蛋白酶 L、组织蛋白酶 B、胰蛋白酶、木瓜蛋白酶和 TMPRSS2[5] 的酶活性。 Decanoyl-RVKR-CMK 通过阻断 Jurkat 淋巴细胞中的包膜糖蛋白前体加工来抑制 HIV-2ROD 复制[1]。
当使用 Decanoyl-RVKR-CMK 时。皮肤鳞状细胞癌细胞系中的 PACE4 活性导致胰岛素样生长因子 1 受体成熟受损，降低其内在酪氨酸激酶活性，并减少肿瘤细胞增殖。两阶段皮肤化学致癌实验以及 CMK 的局部应用表明，癸酰基-RVKR-CMK 显着降低了肿瘤发生率、肿瘤多样性和转移，表明野生型和 PACE4 转基因小鼠的肿瘤进展显着延迟< sup>[6].
| Cas No. | 150113-99-8 | SDF | |
| 别名 | Furin Inhibitor I | ||
| 化学名 | (2S,5R,8R,11S)-5-(4-aminobutyl)-2,11-bis(3-((diaminomethylene)amino)propyl)-8-isopropyl-4,7,10,13-tetraoxo-3,6,9,12-tetraazatricosan-1-oyl chloride | ||
| Canonical SMILES | ClC([C@H](CCC/N=C(N)\N)NC([C@@H](CCCCN)NC([C@@H](C(C)C)NC([C@H](CCC/N=C(N)\N)NC(CCCCCCCCCC)=O)=O)=O)=O)=O | ||
| 分子式 | C34H66ClN11O5 | 分子量 | 744.42 |
| 溶解度 | 33mg/mL in ethanol, or in DMSO, or in DMF | 储存条件 | Store at -20°C |
| General tips | 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 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.3433 mL | 6.7166 mL | 13.4333 mL |
| 5 mM | 0.2687 mL | 1.3433 mL | 2.6867 mL |
| 10 mM | 0.1343 mL | 0.6717 mL | 1.3433 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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Proprotein convertases blockage up-regulates specifically metallothioneins coding genes in human colon cancer stem cells
Biochim Biophys Acta Mol Cell Res2021 Mar;1868(3):118912.PMID: 33249002DOI: 10.1016/j.bbamcr.2020.118912
Despite continuous exertion made, colon cancer still represents a major health problem and its incidence continues being high worldwide. There is growing evidence in support of the cancer stem cells (CSCs) being central in the initiation of this cancer, and CSCs have been the focus of various studies for the identification of new ways of treatment. Lately, the proprotein convertases (PCs) were reported to regulate the maturation and expression of various molecules involved in the malignant phenotype of colon cancer cells, however, the identity of the molecules regulated by these serine proteases in CSCs is unknown. In this study, we used the general PCs inhibitor, the Decanoyl-RVKR-chloromethylketone (Decanoyl-RVKR-CMK) that inhibits all the PCs found in the secretory pathway, and analyzed its effect on CSCs using RNA-seq analysis. Remarkably, from the only 9 up-regulated genes in the human SW620-derived sphere-forming cells, we identified 7 of the 11 human metallothioneins, all of them localized on chromosome 16, and zinc related proteins as downstream effectors of the PCs. The importance of these molecules in the regulation of cell proliferation, differentiation and chemoresistance, and their reported potential tumor suppressor role and loss in colon cancer patients associated with worse prognosis, suggests that targeting PCs in the control of the malignant phenotype of CSCs is a new potential therapeutic strategy in colon cancer.
Mature pig oligodendrocytes rapidly process human recombinant pro-nerve growth factor and do not undergo cell death
J Neurochem2006 Jul;98(2):506-17.PMID: 16805842DOI: 10.1111/j.1471-4159.2006.03891.x
The neurotrophin family with its first member, nerve growth factor (NGF), binds two classes of receptors, more specifically to Trk receptors and to a shared p75NTR receptor. It has been shown that proNGF rather than NGF is predominant in the mature central nervous system. A recent finding indicated that a furin-resistant proNGF preferentially binds to p75NTR, initiating a pro-apoptotic cascade even in the presence of TrkA. In this context, rodent oligodendrocytes were reported to undergo cell death when exposed to proNGF. We have investigated the effect of a non-mutated 32 kDa human recombinant proNGF (rhproNGF) on cultured pig oligodendrocytes which express TrkA, p75NTR and sortilin. Pig oligodendrocytes respond to rhproNGF (50 ng/mL) with an enhanced regeneration of their processes as already observed for NGF. Activity of mitogen-activated protein kinase (MAPK), which plays an important role in oligodendroglial process formation, was increased even when rhproNGF processing was inhibited by the furin inhibitor Decanoyl-RVKR-CMK. Similarly, a cleavage-resistant proNGF (R-1G) activated MAPK and promoted oligodendroglial process regeneration. High concentrations of rhproNGF (300 ng/mL) did not induce cell death. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis and Western blotting revealed that oligodendrocytes process rhproNGF to NGF. NGF was detected in Western blots of oligodendroglial lysates already 10 min after rhproNGF exposure, followed by a release of NGF into the culture medium. Indirect evidence indicates that rhproNGF processing occurs via an endocytotic route.
Mutation of the f-protein cleavage site of avian paramyxovirus type 7 results in furin cleavage, fusion promotion, and increased replication in vitro but not increased replication, tissue tropism, or virulence in chickens
J Virol2012 Apr;86(7):3828-38.PMID: 22258248DOI: 10.1128/JVI.06765-11
We constructed a reverse genetics system for avian paramyxovirus serotype 7 (APMV-7) to investigate the role of the fusion F glycoprotein in tissue tropism and virulence. The AMPV-7 F protein has a single basic residue arginine (R) at position -1 in the F cleavage site sequence and also is unusual in having alanine at position +2 (LPSSR¡ýFA) (underlining indicates the basic amino acids at the F protein cleavage site, and the arrow indicates the site of cleavage.). APMV-7 does not form syncytia or plaques in cell culture, but its replication in vitro does not depend on, and is not increased by, added protease. Two mutants were successfully recovered in which the cleavage site was modified to mimic sites that are found in virulent Newcastle disease virus isolates and to contain 4 or 5 basic residues as well as isoleucine in the +2 position: (RRQKR¡ýFI) or (RRKKR¡ýFI), named Fcs-4B or Fcs-5B, respectively. In cell culture, one of the mutants, Fcs-5B, formed protease-independent syncytia and grew to 10-fold-higher titers compared to the parent and Fcs-4B viruses. This indicated the importance of the single additional basic residue (K) at position -3. Syncytium formation and virus yield of the Fcs-5B virus was impaired by the furin inhibitor decanoyl-RVKR-CMK, whereas parental APMV-7 was not affected. APMV-7 is avirulent in chickens and is limited in tropism to the upper respiratory tract of 1-day-old and 2-week-old chickens, and these characteristics were unchanged for the two mutant viruses. Thus, the acquisition of furin cleavability by APMV-7 resulted in syncytium formation and increased virus yield in vitro but did not alter virus yield, tropism, or virulence in chickens.
A prohormone convertase cleavage site within a predicted alpha-helix mediates sorting of the neuronal and endocrine polypeptide VGF into the regulated secretory pathway
J Biol Chem2005 Dec 16;280(50):41595-608.PMID: 16221685DOI: 10.1074/jbc.M509122200
Distinct intracellular pathways are involved in regulated and constitutive protein secretion from neuronal and endocrine cells, yet the peptide signals and molecular mechanisms responsible for targeting and retention of soluble proteins in secretory granules are incompletely understood. By using confocal microscopy and subcellular fractionation, we examined trafficking of the neuronal and endocrine peptide precursor VGF that is stored in large dense core vesicles and undergoes regulated secretion. VGF cofractionated with secretory vesicle membranes but was not detected in detergent-resistant lipid rafts. Deletional analysis using epitope-tagged VGF suggested that the C-terminal 73-amino acid fragment of VGF, containing two predicted alpha-helical loops and four potential prohormone convertase (PC) cleavage sites, was necessary and sufficient with an N-terminal signal peptide-containing domain, for large dense core vesicle sorting and regulated secretion from PC12 and INS-1 cells. Further transfection analysis identified the sorting sequence as a compact C-terminal alpha-helix and embedded 564RRR566 PC cleavage site; mutation of the 564RRR566 PC site in VGF-(1-65): GFP:VGF-(545-617) blocked regulated secretion, whereas disruption of the alpha-helix had no effect. Mutation of the adjacent 567HFHH570 motif, a charged region that might enhance PC cleavage in acidic environments, also blocked regulated release. Finally, inhibition of PC cleavage in PC12 cells using the membrane-permeable synthetic peptide chloromethyl ketone (decanoyl-RVKR-CMK) blocked regulated secretion of VGF. Our studies define a critical RRR-containing C-terminal domain that targets VGF into the regulated pathway in neuronal PC12 and endocrine INS-1 cells, providing additional support for the proposed role that PCs and their cleavage sites play in regulated peptide secretion.