Polymyxin B nonapeptide
(Synonyms: 多粘菌素B九肽) 目录号 : GC33369
Polymyxin B nonapeptide, as a cationic cyclic peptide derived from polymyxin B, can increase the permeability of the outer membrane (OM) of GNB toward hydrophobic antibiotics probably.
Cas No.:86408-36-8
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >98.00%
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- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Four ESBL-producing E. coli isolates |
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Preparation Method |
PMBN (5 µg/ml), was examined in combination with miconazole and DETA/NO and the viability evaluated in all four ESBL isolates and in J96δhmp. |
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Reaction Conditions |
5 µg/ml; 24h |
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Applications |
PMBN and miconazole in combination slightly reduced the bacterial growth. DETA/NO and PMBN in combination showed full recovery of growth after 24 h. |
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References: [1] Bang CS, et al. The antibacterial effect of nitric oxide against ESBL-producing uropathogenic E. coli is improved by combination with miconazole and polymyxin B nonapeptide. BMC Microbiol. 2014 Mar 14;14:65. |
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Polymyxin B nonapeptide, as a cationic cyclic peptide derived from polymyxin B, can increase the permeability of the outer membrane (OM) of GNB toward hydrophobic antibiotics probably. Polymyxin B nonapeptide has less toxicity and no bactericidal activity[1].
In vitro cytotoxicity experiments, Polymyxin B nonapeptide show IC50 with >32 μg/ml in FBS, MDCK, HaCaT and HEK 239 cell lines[2]. In vitro experiment it shown that at 32 and 16 μg/ml Polymyxin B nonapeptide with > 4 μg/ml azithromycin can kill bacterial. While at 8, 4 and 2 μg/ml Polymyxin B nonapeptide in combination with > 8 μg/ml azithromycin also caused same effect[1]. In vitro, smooth Salmonella typhimurium had a binding capacity of ca. 6 nmol of Polymyxin B nonapeptide per mg (dry weight) of bacteria, which is equivalent to ca. 1 X 106 to 2 X 106 molecules of Polymyxin B nonapeptide per single cell. The binding was of relatively high affinity with Kd of 1.3 μM. In addition, the isolated outer membrane of S. typhimurium bound ca. 100 nmol of Polymyxin B nonapeptide per mg of outer membrane protein with Kd of 1.1 μM, whereas the cytoplasmic membrane bound 9 to 10 times less[3]. In vitro, with 4 μg/ml Polymyxin B nonapeptide can increase electrotransformation efficiency of the growth of uropathogenic E. coli[4].
In vivo experiment it shown that polymyxin B nonapeptide with 1.5 and 3.0 mg/kg did not exhibit the neuromuscular blocking, neurotoxic, or nephrotoxic effects[5].
Al-Marzooq F, et al. Discerning the role of polymyxin B nonapeptide in restoring the antibacterial activity of azithromycin against antibiotic-resistant Escherichia coli. Front Microbiol. 2022 Sep 21;13:998671.
[2] Khazandi M, et al. In vitro Antimicrobial Activity of Robenidine, Ethylenediaminetetraacetic Acid and Polymyxin B Nonapeptide Against Important Human and Veterinary Pathogens. Front Microbiol. 2019 Apr 25;10:837.
[3] Vaara M, et al. Binding of polymyxin B nonapeptide to gram-negative bacteria. Antimicrob Agents Chemother. 1985 Apr;27(4):548-54.
[4] Qin J, et al. A method for increasing electroporation competence of Gram-negative clinical isolates by polymyxin B nonapeptide. Sci Rep. 2022 Jul 8;12(1):11629.
[5] Danner RL, et al. Purification, toxicity, and antiendotoxin activity of polymyxin B nonapeptide. Antimicrob Agents Chemother. 1989 Sep;33(9):1428-34.
References:
多粘菌素 B 九肽作为衍生自多粘菌素 B 的阳离子环肽，可能会增加 GNB 外膜 (OM) 对疏水性抗生素的通透性。多粘菌素B九肽毒性较小，无杀菌活性[1]。
在体外细胞毒性实验中，多粘菌素 B 九肽在 FBS、MDCK、HaCaT 和 HEK 239 细胞系[2] 中的 IC50 为 >32 μg/ml。体外实验表明，在 32 和 16 μg/ml 的多粘菌素 B 九肽中，具有 >； 4 μg/ml 阿奇霉素可杀灭细菌。在 8、4 和 2 μg/ml 多粘菌素 B 九肽与 > 组合时； 8 μg/ml 阿奇霉素也有同样的作用[1]。在体外，光滑的鼠伤寒沙门氏菌具有约 10 的结合能力。每 mg（干重）细菌含 6 nmol 多粘菌素 B 九肽，相当于 ca。每个单细胞 1 X 106 至 2 X 106 个多粘菌素 B 九肽分子。该结合具有相对较高的亲和力，Kd 为 1.3 μM。此外，鼠伤寒沙门氏菌的分离外膜结合约。每 mg 外膜蛋白加入 100 nmol 多粘菌素 B 九肽，Kd 为 1.1 μM，而胞质膜结合减少 9 至 10 倍[3]。在体外，4 μg/ml Polymyxin B nonapeptide 可提高尿路致病性大肠杆菌生长的电转化效率[4]。
体内实验表明，1.5 和 3.0 mg/kg 的多粘菌素 B 九肽没有表现出神经肌肉阻滞、神经毒性或肾毒性作用[5]。
Al-Marzooq F 等人。辨别多粘菌素 B 九肽在恢复阿奇霉素对抗生素耐药大肠杆菌的抗菌活性中的作用。前微生物。 2022 年 9 月 21 日；13:998671。
[2] Khazandi M 等人。 Robenidine、乙二胺四乙酸和多粘菌素 B 九肽对重要人类和兽医病原体的体外抗菌活性。前微生物。 2019 年 4 月 25 日；10:837。
[3] Vaara M 等人。多粘菌素 B 九肽与革兰氏阴性菌的结合。抗菌剂化疗。 1985 年 4 月；27(4):548-54。
[4] 秦杰等。一种通过多粘菌素 B 九肽提高革兰氏阴性临床分离株电穿孔能力的方法。科学报告 2022 年 7 月 8 日；12(1):11629。
[5] Danner RL 等人。多粘菌素 B 九肽的纯化、毒性和抗内毒素活性。抗菌剂化疗。 1989 年 9 月；33(9):1428-34。
| Cas No. | 86408-36-8 | SDF | |
| 别名 | 多粘菌素B九肽 | ||
| Canonical SMILES | Thr-{Dab}-{Dab}-{Dab}-{Dab}-d-Phe-Leu-{Dab}-{Dab}-Thr (Lactam: Dab3-Thr9) | ||
| 分子式 | C43H74N14O11 | 分子量 | 963.13 |
| 溶解度 | Water: ≥ 100 mg/mL (103.83 mM); DMSO: 16.67 mg/mL (17.31 mM) | 储存条件 | Store at -20°C, protect from light |
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.0383 mL | 5.1914 mL | 10.3828 mL |
| 5 mM | 0.2077 mL | 1.0383 mL | 2.0766 mL |
| 10 mM | 0.1038 mL | 0.5191 mL | 1.0383 mL |
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动物平均体重 | g | |
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动物数量 | 只 |
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2.
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Discerning the role of Polymyxin B nonapeptide in restoring the antibacterial activity of azithromycin against antibiotic-resistant Escherichia coli
Front Microbiol 2022 Sep 21;13:998671.PMID:36212888DOI:10.3389/fmicb.2022.998671.
Antimicrobial resistance is a global public health threat. Antibiotic development pipeline has few new drugs; therefore, using antibiotic adjuvants has been envisioned as a successful method to preserve existing medications to fight multidrug-resistant (MDR) pathogens. In this study, we investigated the synergistic effect of a polymyxin derivative known as Polymyxin B nonapeptide (PMBN) with azithromycin (AZT). A total of 54 Escherichia coli strains were first characterized for macrolide resistance genes, and susceptibility to different antibiotics, including AZT. A subset of 24 strains was then selected for synergy testing by the checkerboard assay. PMBN was able to re-sensitize the bacteria to AZT, even in strains with high minimum inhibitory concentrations (MIC: 32 to ≥128 μg/ml) for AZT, and in strains resistant to the last resort drugs such as colistin and meropenem. The fractional inhibitory concentration index was lower than 0.5, demonstrating that PMBN and AZT combinations had a synergistic effect. The combinations worked efficiently in strains carrying mphA gene encoding macrolide phosphotransferase which can cause macrolide inactivation. However, the combinations were inactive in strains having an additional ermB gene encoding macrolide methylase which causes ribosomal drug target alteration. Killing kinetics study showed a significant reduction of bacterial growth after 6 h of treatment with complete killing achieved after 24 h. Transmission electron microscopy showed morphological alterations in the bacteria treated with PMBN alone or in combination with AZT, with evidence of damage to the outer membrane. These results suggested that PMBN acted by increasing the permeability of bacterial outer membrane to AZT, which was also evident using a fluorometric assay. Using multiple antimicrobial agents could therefore be a promising strategy in the eradication of MDR bacteria. PMBN is a good candidate for use with other antibiotics to potentiate their activity, but further studies are required in vivo. This will significantly contribute to resolving antimicrobial resistance crisis.
Purification, toxicity, and antiendotoxin activity of Polymyxin B nonapeptide
Antimicrob Agents Chemother 1989 Sep;33(9):1428-34.PMID:2554795DOI:10.1128/AAC.33.9.1428.
Polymyxin B, a relatively toxic antibiotic, has potent endotoxin-neutralizing properties that may be beneficial as adjunctive therapy in gram-negative sepsis. Polymyxin B nonapeptide (deacylated polymyxin B) is devoid of antibiotic activity but retains the capacity to disorganize the outer membrane of gram-negative bacteria. To evaluate the potential therapeutic usefulness of this derivative, we produced purified Polymyxin B nonapeptide, tested its in vivo toxicity in animals, and evaluated its in vitro antiendotoxin activity. Effectiveness as an antiendotoxin agent was assessed by examining the ability of Polymyxin B nonapeptide to block the enhanced release of toxic oxygen radicals induced by lipopolysaccharide in human neutrophils (priming). In vivo, at doses of 1.5 and 3.0 mg/kg, Polymyxin B nonapeptide did not exhibit the neuromuscular blocking, neurotoxic, or nephrotoxic effects that were observed with polymyxin B sulfate. Both polymyxin B and Polymyxin B nonapeptide inhibited lipopolysaccharide-induced neutrophil priming in a concentration-dependent manner, but the parent compound, polymyxin B, was 63 times more effective on a weight basis. The inhibitory activity of both compounds, however, diminished rapidly when they were added after the start of the lipopolysaccharide-neutrophil incubation. We conclude that Polymyxin B nonapeptide is less toxic than polymyxin B and, at the doses tested, lacks the neurotoxicity and nephrotoxicity of the parent compound. Polymyxin B nonapeptide retains the antiendotoxin activity of polymyxin B but is much less potent. The findings suggest that these compounds block an early step in the neutrophil priming process, possibly lipopolysaccharide attachment to or insertion into the neutrophil membrane.
A method for increasing electroporation competence of Gram-negative clinical isolates by Polymyxin B nonapeptide
Sci Rep 2022 Jul 8;12(1):11629.PMID:35804085DOI:10.1038/s41598-022-15997-8.
The study of clinically relevant bacterial pathogens relies on molecular and genetic approaches. However, the generally low transformation frequency among natural isolates poses technical hurdles to widely applying common methods in molecular biology, including transformation of large constructs, chromosomal genetic manipulation, and dense mutant library construction. Here we demonstrate that culturing clinical isolates in the presence of Polymyxin B nonapeptide (PMBN) improves their transformation frequency via electroporation by up to 100-fold in a dose-dependent and reversible manner. The effect was observed for PMBN-binding uropathogenic Escherichia coli (UPEC) and Salmonella enterica strains but not naturally polymyxin resistant Proteus mirabilis. Using our PMBN electroporation method we show efficient delivery of large plasmid constructs into UPEC, which otherwise failed using a conventional electroporation protocol. Moreover, we show a fivefold increase in the yield of engineered mutant colonies obtained in S. enterica with the widely used lambda-Red recombineering method, when cells are cultured in the presence of PMBN. Lastly, we demonstrate that PMBN treatment can enhance the delivery of DNA-transposase complexes into UPEC and increase transposon mutant yield by eightfold when constructing Transposon Insertion Sequencing (TIS) libraries. Therefore, PMBN can be used as a powerful electropermeabilisation adjuvant to aid the delivery of DNA and DNA-protein complexes into clinically important bacteria.
Polymyxin B and Polymyxin B nonapeptide alter cytoplasmic membrane permeability in Escherichia coli
J Antimicrob Chemother 1986 Nov;18(5):557-63.PMID:3027012DOI:10.1093/jac/18.5.557.
The effects of polymyxin B and Polymyxin B nonapeptide (PMBN) on the permeability of the Escherichia coli cytoplasmic membrane were investigated. Both compounds caused loss of free amino acids, uracil and K+ from E. coli. The rates of loss promoted by polymyxin B were one and a half to two-fold greater than those caused by PMBN. Although PMBN mediated loss of low molecular weight substances from E. coli, it was not bactericidal. In contrast, polymyxin B treated E. coli lysed and rapidly lost viability. We suggest that the bactericidal activity of polymyxin B may be related to its previously reported ability to release cytoplasmic proteins from bacteria.
Effects of polymyxin B sulfate and Polymyxin B nonapeptide on growth and permeability of the yeast Saccharomyces cerevisiae
Mol Gen Genet 1985;199(3):401-5.PMID:2993791DOI:10.1007/BF00330749.
Polymyxin B, a toxic, membrane-affecting antibiotic, can be rendered harmless to yeast cells by enzymatic removal of its fatty acyl moiety. The remaining cyclic peptide portion, Polymyxin B nonapeptide, has no significant effect on growth and viability but it drastically reduces mating efficiency. In addition, the cyclic peptide enhances sensitivity of cells to several drugs, presumably by increasing membrane permeability. Mutants resistant to polymyxin B are simultaneously less responsive to the combination of the nonapeptide and the drugs. This indicates that the peptide portion of polymyxin B is the moiety responsible for the permeability changes. The resistance is inherited as a simple recessive trait. The mutation has been mapped to chromosome XV of Saccharomyces cerevisiae.