Pam3CSK4
(Synonyms: Pam3Cys-Ser-(Lys)4) 目录号 : GC10273
Pam3CSK4 (Pam3CysSerLys4) 是一种合成的三酰化脂肽 (LP) 和 TLR2/TLR1 配体激动剂，EC50 为 0.47 ng/mL。
Cas No.:112208-00-1
Sample solution is provided at 25 µL, 10mM.
Pam3CSK4 (Pam3CysSerLys4) is a synthetic triacylated lipopeptide (LP) and a TLR2/TLR1 ligand agonist with an EC50 of 0.47 ng/mL[10]. Pam3CSK4 mimics the acylated amino terminus of bacterial LPs. Bacterial LPs are a family of pro-inflammatory cell wall components found in both Gram-positive and Gram-negative bacteria. These bacterial LPs are recognized by TLR2, a receptor that plays a pivotal role in detecting a diverse range of pathogen-associated molecular patterns (PAMPs).Recognition of Pam3CSK4, a triacylated LP, is mediated by TLR2 which cooperates with TLR1 through their cytoplasmic domain to induce the signaling cascade leading to the activation of NF-κB [1].
In HepaRG cells, Pam3CSK4 shows a broad antiviral activity against HBV. Pam3CSK4 is a well-characterized and specific TLR1/2 ligand[2]. The anti-HBV effect of Pam4CSK4 is dependent on TLR1/2 and its adaptor MyD88, targeting TLR1 or TLR6 with specific siRNA do affect the feed-forward expression of TLR2 in the context of Pam3CSK4 activation ; but invalidation of TLR1 led to a reduced inhibitory effect of Pam3CSK4. The low functional level of TLR2 was sufficient to produce the therapeutic effect of Pam3CSK4[4,5].Thus confirming that Pam3CSK4 likely signals through TLR1/2 heterodimers in hepatocytes[3].
Activation of TLR2/1 heterodimers by Pam3CSK4 mediates pain and itching, whereas activation of TLR2/6 heterodimers by lipoteichoic acid (LTA) or yeast glycan leads to itching[9]. As a cutaneous regulator, IL-13 activates sensory neurons and participates in the initiation of AD and itch[7]. IL-13 alone did not lead to a significant increase in scratching number，TLR2 promotes IL-13 signaling in sensory neurons. Innate TLR2 signaling not only promotes itch and pain but convert transient TH2 cell-mediated dermatitis into persistent inflammation which is linked to chronic human AD[8]. In mice pretreated with Pam3CSK4, IL-13 injection caused approximately twice as many scratches as vehicle injection[6]. Pam3CSK4 enhances IL-13-induced calcium transient in sensory neurons and elevates IL-13-induced Itch-like behaviours in mice.
[1]: Ozinsky A, Underhill DM, et,al.The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between toll-like receptors. Proc Natl Acad Sci U S A. 2000 Dec 5;97(25):13766-71. doi: 10.1073/pnas.250476497. PMID: 11095740; PMCID: PMC17650.
[2]: Lucifora J, Xia Y, et,al. Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA. Science. 2014 Mar 14;343(6176):1221-8. doi: 10.1126/science.1243462. Epub 2014 Feb 20. PMID: 24557838; PMCID: PMC6309542.
[3]: K. Visvanathan, N.A. Skinner, et al. Regulation of Toll-like receptor-2 expression in chronic hepatitis B by the precore protein. Hepatology. doi:10.1002. 2006.
[4]: Xiao S, Lu Z, et,al. Innate immune regulates cutaneous sensory IL-13 receptor alpha 2 to promote atopic dermatitis. Brain Behav Immun. 2021 Nov;98:28-39. doi: 10.1016/j.bbi.2021.08.211. Epub 2021 Aug 13. PMID: 34391816.
[5]: Erickson S, Heul AV, et,al. New and emerging treatments for inflammatory itch. Ann Allergy Asthma Immunol. 2021 Jan;126(1):13-20. doi: 10.1016/j.anai.2020.05.028. Epub 2020 Jun 1. PMID: 32497711.
[6]: Oetjen LK, Mack MR, et,al. Sensory Neurons Co-opt Classical Immune Signaling Pathways to Mediate Chronic Itch. Cell. 2017 Sep 21;171(1):217-228.e13. doi: 10.1016/j.cell.2017.08.006. Epub 2017 Sep 7. PMID: 28890086; PMCID: PMC5658016.
[7]: Kaesler S, Volz T, et,al. Toll-like receptor 2 ligands promote chronic atopic dermatitis through IL-4-mediated suppression of IL-10. J Allergy Clin Immunol. 2014 Jul;134(1):92-9. doi: 10.1016/j.jaci.2014.02.017. Epub 2014 Apr 1. PMID: 24698321.
[8]: Liu T, Gao YJ, et,al. Emerging role of Toll-like receptors in the control of pain and itch. Neurosci Bull. 2012 Apr;28(2):131-44. doi: 10.1007/s12264-012-1219-5. PMID: 22466124; PMCID: PMC3347759.
[9]:Wang TT, Xu XY, et,al. Activation of Different Heterodimers of TLR2 Distinctly Mediates Pain and Itch. Neuroscience. 2020 Mar 1;429:245-255. doi: 10.1016/j.neuroscience.2020.01.010. Epub 2020 Jan 16. PMID: 31954829.
[10]:Irvine KL, Hopkins LJ, Gangloff M, Bryant CE. The molecular basis for recognition of bacterial ligands at equine TLR2, TLR1 and TLR6. Vet Res. 2013 Jul 4;44(1):50. doi: 10.1186/1297-9716-44-50. PMID: 23826682; PMCID: PMC3716717.
Pam3CSK4 (Pam3CysSerLys4) 是一种合成的三酰化脂肽 (LP) 和 TLR2/TLR1 配体激动剂，EC50 为 0.47 ng/mL[10]。 Pam3CSK4 模仿细菌 LP 的酰化氨基末端。细菌 LP 是在革兰氏阳性和革兰氏阴性细菌中发现的促炎细胞壁成分家族。这些细菌 LP 被 TLR2 识别，TLR2 是一种在检测多种病原体相关分子模式 (PAMP) 中起关键作用的受体。Pam3CSK4（一种三酰化 LP）的识别由 TLR2 介导，TLR2 通过其细胞质结构域与 TLR1 合作诱导导致 NF-κB 激活的信号级联反应[1]。
在 HepaRG 细胞中，Pam3CSK4 显示出针对 HBV 的广泛抗病毒活性。 Pam3CSK4 是一种充分表征的特异性 TLR1/2 配体[2]。 Pam4CSK4 的抗 HBV 作用依赖于 TLR1/2 及其衔接子 MyD88，靶向 TLR1 或 TLR6 的特异性 siRNA 确实会在 Pam3CSK4 激活的背景下影响 TLR2 的前馈表达；但 TLR1 的失效导致 Pam3CSK4 的抑制作用降低。 TLR2 的低功能水平足以产生 Pam3CSK4[4,5] 的治疗效果。从而证实 Pam3CSK4 可能通过肝细胞中的 TLR1/2 异二聚体发出信号[3].
Pam3CSK4 激活 TLR2/1 异二聚体可介导疼痛和瘙痒，而脂磷壁酸 (LTA) 或酵母聚糖激活 TLR2/6 异二聚体可导致瘙痒[9]。作为皮肤调节剂，IL-13 激活感觉神经元并参与 AD 和瘙痒的启动[7]。单独的 IL-13 不会导致抓挠次数的显着增加，TLR2 促进感觉神经元中的 IL-13 信号传导。先天性 TLR2 信号不仅会促进瘙痒和疼痛，还会将短暂的 TH2 细胞介导的皮炎转化为与慢性人类 AD[8] 相关的持续性炎症。在用 Pam3CSK4 预处理的小鼠中，IL-13 注射引起的划痕数量大约是载体注射的两倍[6]。 Pam3CSK4 增强 IL-13 诱导的感觉神经元钙瞬变，并增强 IL-13 诱导的小鼠瘙痒样行为。
| Cell experiment [1]: | |
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Cell lines |
HepaRG cell |
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Preparation Method |
Indicated differentiated HepaRG cell lines were infected with HBV with 100 vge/cell. At day-7 post-infection , cells were treated twice with either Pam3CSK4 (100 ng/mL), IFNα (500 IU/mL), RG7834 (0.1 μM), or the nucleoside analogue lamivudine (3 TC; 1 μM) for a total exposure time of 6 days. Cells and supernatants were harvested for analyses at day-13 post-infection. |
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Reaction Conditions |
100 ng/mL; 6 days |
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Applications |
Pam3CSK4 was capable to inhibit HBV (genotype D) replication in dHepaRG cells. The levels of all HBV parameters analyzed, (including intracellular total HBV RNAs, viremia, and secretion of viral antigens) were significantly reduced even at very low concentration of Pam3CSK4. Pam3CSK4 was also capable to inhibit the replication of another HBV genotype (genotype C). thus suggesting a broad antiviral activity against HBV. Pam3CSK4 as a TLR2 agonist, is a potent anti-HBV agents. Pam3CSK4 inhibits RNA accumulation by both impairing HBV transcription and RNA stability. Pam3CSK4 decreases also cccDNA level via a FEN-1-dependent mechanism. |
| Animal experiment [2]: | |
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Animal models |
C57 BL/6 mice |
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Preparation Method |
To analyse the influence of TLR2 heterodimer activation on IL-13-induced itch-like behaviour, C57 BL/6 mice were pre-administered with Pam3CSK4 (20 μg/100 μl sterilized water) by gavage every three days for 3 times . In another experiment, FSL-1 (0.3 mg/kg, 60 μl sterilized water) was intraperitoneally injected 14 h before IL-13 injection. Control mice were given sterilized water only. All of these mice were then intradermally injected with IL-13 (1 μg/10 μl) or vehicle into the left cheek. All the mice were video-recorded for 1 h for the analysis of scratching bouts. |
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Dosage form |
Pam3CSK4 (20 μg/100 μl sterilized water) by gavage every three days for 3 times |
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Applications |
As a cutaneous regulator, IL-13 activates sensory neurons and participates in the initiation of AD and itch. Pam3CSK4 enhances IL-13-induced calcium transient in sensory neurons and elevates IL-13-induced Itch-like behaviours in mice。 |
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References: [1]. Desmares M, Delphin M,et,al. Insights on the antiviral mechanisms of action of the TLR1/2 agonist Pam3CSK4 in hepatitis B virus (HBV)-infected hepatocytes. Antiviral Res. 2022 Aug 10;206:105386. doi: 10.1016/j.antiviral.2022.105386. Epub ahead of print. PMID: 35963549. [2]. Xiao S, Lu Z, et,al. Innate immune regulates cutaneous sensory IL-13 receptor alpha 2 to promote atopic dermatitis. Brain Behav Immun. 2021 Nov;98:28-39. doi: 10.1016/j.bbi.2021.08.211. Epub 2021 Aug 13. PMID: 34391816. |
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| Cas No. | 112208-00-1 | SDF | |
| 别名 | Pam3Cys-Ser-(Lys)4 | ||
| 化学名 | (2S,3Z,5S,6Z,8S,9Z,11S,12Z,14S,15Z,17R)-2,5,8,11-tetrakis(4-aminobutyl)-4,7,10,13,16-pentahydroxy-17-((Z)-(1-hydroxyhexadecylidene)amino)-14-(hydroxymethyl)-24-oxo-21-(palmitoyloxy)-23-oxa-19-thia-3,6,9,12,15-pentaazanonatriaconta-3,6,9,12,15-pentaen-1-oi | ||
| Canonical SMILES | CCCCCCCCCCCCCCC/C(O)=N/[C@@](/C(O)=N/[C@@](/C(O)=N/[C@@](/C(O)=N/[C@@](/C(O)=N/[C@@](/C(O)=N/[C@@](C(O)=O)([H])CCCCN)([H])CCCCN)([H])CCCCN)([H])CCCCN)([H])CO)([H])CSCC(OC(CCCCCCCCCCCCCCC)=O)([H])COC(CCCCCCCCCCCCCCC)=O | ||
| 分子式 | C81H156N10O13S | 分子量 | 1510.24 |
| 溶解度 | 50 mg/mL in DMSO ( Need ultrasonic); 16.67 mg/mL in Water ( Needultrasonic) | 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。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 0.6621 mL | 3.3107 mL | 6.6215 mL |
| 5 mM | 0.1324 mL | 0.6621 mL | 1.3243 mL |
| 10 mM | 0.0662 mL | 0.3311 mL | 0.6621 mL |
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| 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 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
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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Quality Control & SDS
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- Purity: >98.00%
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