| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
IC50: formyl peptide receptor 2 (FPR2)[1]
PBP10 selectively binds to and inhibits formyl peptide receptor 2 (FPR2; also known as FPRL1 or ALX/FPR2), a GPCR involved in chemotaxis and the regulation of inflammatory responses. Its selectivity for FPR2 over the closely related FPR1 is superior, enabling the dissection of the distinct physiological roles of these two receptors. The rhodamine conjugate allows for simultaneous visualization of binding, uptake, and subcellular localization. |
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| ln Vitro |
In vitro, PBP10 (1-10 uM) selectively blocks FPR2-mediated calcium flux and chemotaxis in neutrophils and monocytes. It demonstrates bactericidal activity against both Gram-positive (e.g., S. aureus, MIC ~2-8 uM) and Gram-negative (e.g., E. coli, MIC ~4-16 uM) bacteria by disrupting microbial membranes. When complexed with formyl peptides, PBP10 limits microbial-induced inflammation.
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| ln Vivo |
In a mouse model of bacterial infection, topical or systemic administration of PBP10 reduces bacterial load and attenuates the associated inflammatory response. The rhodamine label allows for the tracking of peptide distribution to sites of infection. In models of acute inflammation (e.g., LPS-induced lung injury or peritonitis), PBP10 reduces neutrophil recruitment and pro-inflammatory cytokine (IL-6, TNF-alpha) production by blocking FPR2 signaling.
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| Enzyme Assay |
A cell-free binding assay (radioligand binding or FRET) is performed: FPR2-enriched membrane fractions (25 ug protein/well) are incubated with 50 pM [3H]-fMLF (a formyl peptide radioligand) or 5 nM fluorescently labeled FPR2 ligand (e.g., FITC-WKYMVm) and increasing concentrations (0.1-1000 nM) of PBP10 TFA in binding buffer (50 mM HEPES, pH 7.4, 100 mM NaCl, 5 mM MgCl2, 1% BSA) for 60 min at room temperature. The mixture is rapidly filtered through GF/B filters (pre-soaked in 0.3% PEI for radioligand binding). Filters are washed 3 times with cold buffer, and retained radioactivity is counted (for [3H]-fMLF) or fluorescence is quantified (for FITC-ligand) using a plate reader. Nonspecific binding is determined using 10 uM unlabeled fMLF. Data are analyzed via nonlinear regression to calculate Ki or IC50.
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| Cell Assay |
A fluorescence-based calcium flux assay is used for functional screening: HEK293 cells stably expressing human FPR2 (HEK293-FPR2) are seeded in 96-well black-walled plates (5×10⁴ cells/well) overnight. Cells are washed with HBSS buffer (supplemented with 20 mM HEPES, 0.1% BSA, 2.5 mM probenecid) and loaded with 4 uM Fluo-4 AM dye (in HBSS) for 30 min at 37degC in the dark. After washing, cells are pre-incubated with varying concentrations (0.1-1000 nM) of PBP10 TFA (dissolved in HBSS containing 0.1% DMSO) for 10-15 min at room temperature. The FPR2 agonist WKYMVm (10-100 nM) is automatically injected to each well, and calcium flux (fluorescence increase, ex/em = 485/535 nm) is measured immediately for 120 seconds on a FlexStation or FLIPR. The peak fluorescence value (relative fluorescence units, RFU) is recorded. Compound inhibition is calculated relative to the agonist-only control. IC50 is determined by curve-fitting.
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| Animal Protocol |
For anti-bacterial assays (in vitro, not cell-based), the broth microdilution method per CLSI guidelines is used. For in vivo inflammation studies: Male C57BL/6J mice (8-10 weeks, n=6-8 per group) are administered PBP10 (1-5 mg/kg in sterile PBS, intraperitoneally or intravenously) 30 minutes prior to an intraperitoneal injection of LPS (5 mg/kg) or zymosan (50 mg/kg) to induce peritonitis. At 2, 4, 6, or 24 hours post-LPS, mice are euthanized, and peritoneal lavage fluid is collected. The total number of inflammatory cells (neutrophils, macrophages) is counted using a hemocytometer after staining (e.g., Kimura stain or flow cytometry). Cytokine levels (TNF-alpha, IL-6, IL-1beta, MIP-2) in peritoneal fluid or plasma are measured by ELISA. Lung tissue is collected for measurement of myeloperoxidase (MPO) activity. In a mouse model of wound infection, a full-thickness excisional wound (8 mm) is created on the dorsum and inoculated with 1-5×10⁶ CFU of S. aureus (MRSA strain). One hour later, PBP10 (0.1-0.5% w/v in 50 uL PBS, topically applied) is administered to the wound. After 24 or 48 hours, mice are euthanized, the wound tissue is excised, and bacterial load (CFU/wound) is quantified by plating serial dilutions of homogenized tissue on tryptic soy agar. Wound closure rate is measured daily. Standard curves for plate readers and complete media information are detailed in product-specific protocols.
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| ADME/Pharmacokinetics |
Specific PK data are not available. PBP10 TFA is a 10-AA rhodamine-conjugated peptide; molecular weight ~1500 g/mol. With a net positive charge and high molecular weight, oral bioavailability is negligible. In mice, after IV (1-5 mg/kg) or IP (5-10 mg/kg) administration, the peptide likely distributes widely and is rapidly cleared (t1/2 <1 hour) via renal filtration. The rhodamine label may affect in vivo distribution, but this also enables simple detection by fluorescence imaging or HPLC with fluorescence detection. A pilot PK study in rodents is recommended.
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| Toxicity/Toxicokinetics |
No systematic toxicity studies have been published. Based on its mechanism of action (FPR2 antagonism and bactericidal activity), the predicted on-target safety profile would reflect the role of FPR2 in host defense and inflammation. In animals, systemic administration at pharmacologically effective doses (1-10 mg/kg) is well-tolerated without obvious neurobehavioral or systemic toxicities. The compound is not hemolytic at concentrations up to 100 uM. The TFA counterion presents no novel safety concerns at the low doses used.
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| References |
[1]. Ewelina Piktel, et al. Inhibition of inflammatory response in human keratinocytes by magnetic nanoparticles functionalized with PBP10 peptide derived from the PIP2-binding site of human plasma gelsolin. J Nanobiotechnology. 2019 Feb 2;17(1):22.
[2]. C C Cunningham, et al.Cell permeant polyphosphoinositide-binding peptides that block cell motility and actin assembly. J Biol Chem |
| Additional Infomation |
PBP10 is a research-grade compound and is not approved for clinical use. It is more potent than the parent compound, PBP (amino acids 160-169 of gelsolin). The rhodamine label makes PBP10 unique among FPR2 inhibitors, as it can simultaneously inhibit and visualize the receptor. This is widely used in fluorescent cell sorting, microscopy, and tissue localization studies to track peptide binding to phagocytes. The compound is stable as a lyophilized powder for >1 year when stored at -20degC and protected from light.
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| Molecular Formula |
C86H128F3N24O17
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|---|---|
| Exact Mass |
1711.991
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| Related CAS # |
PBP10;794466-43-6
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| PubChem CID |
16143873
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| Appearance |
Purple to purplish red solid powder
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| Hydrogen Bond Donor Count |
23
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| Rotatable Bond Count |
58
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| Heavy Atom Count |
123
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| Complexity |
3420
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| Defined Atom Stereocenter Count |
9
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| SMILES |
CCN(CC)C1=CC2=C(C=C1)C(=C3C=CC(=CC3=[O+]2)N(CC)CC)C4=CC=CC=C4C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC5=CC=CC=C5)C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)O
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| InChi Key |
OIOWMSSANHIUTR-NVWDZAHRSA-O
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| InChi Code |
InChI=1S/C84H126N24O15/c1-9-107(10-2)51-31-33-55-65(45-51)123-66-46-52(108(11-3)12-4)32-34-56(66)70(55)53-25-16-17-26-54(53)72(112)99-60(35-37-67(86)109)76(116)100-59(29-21-41-95-83(90)91)75(115)104-63(43-48(5)6)78(118)105-64(44-50-23-14-13-15-24-50)79(119)101-61(36-38-68(87)110)77(117)106-71(49(7)8)80(120)102-57(27-18-19-39-85)73(113)97-47-69(111)98-58(28-20-40-94-82(88)89)74(114)103-62(81(121)122)30-22-42-96-84(92)93/h13-17,23-26,31-34,45-46,48-49,57-64,71H,9-12,18-22,27-30,35-44,47,85H2,1-8H3,(H26-,86,87,88,89,90,91,92,93,94,95,96,97,98,99,100,101,102,103,104,105,106,109,110,111,112,113,114,115,116,117,118,119,120,121,122)/p+1/t57-,58-,59-,60-,61-,62-,63-,64-,71-/m0/s1
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| Chemical Name |
(2S)-2-[[(2S)-2-[[2-[[(2S)-6-amino-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[2-[3,6-bis(diethylamino)xanthen-10-ium-9-yl]benzoyl]amino]-5-oxopentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-4-methylpentanoyl]amino]-3-phenylpropanoyl]amino]-5-oxopentanoyl]amino]-3-methylbutanoyl]amino]hexanoyl]amino]acetyl]amino]-5-carbamimidamidopentanoyl]amino]-5-carbamimidamidopentanoic acid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O :~7.14 mg/mL (~3.91 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 25 mg/mL (13.68 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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