Closthioamide
目录号 : GC32291
Closthioamide是细菌DNA促旋酶的一个有效抑制剂，其对Ec，MRSA，VRE，Mv的最小抑菌浓度分别是9.00μM，0.58μM，0.58μM和72.03μM。
Cas No.:1227367-59-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Cell experiment: | The MIC is determined for the laboratory strains as follows. The assays are performed in sterile 96-well polystyrene microplates in 200 mL of cation-adjusted Mueller-Hinton (MH) broth. A serial dilution of the compound to test is made in 100 mL of medium and inoculated with 100 mL of fresh cell culture containing ~105 cfu/mL and incubated at 37°C for 20 h. The MIC is defined as the lowest concentration of antibiotic that inhibit visible growth[1]. |
References: [1]. Chiriac AI, et al. Mode of action of closthioamide: the first member of the polythioamide class of bacterial DNA gyrase inhibitors. J AntimicrobChemother. 2015Sep;70(9):2576-88. | |
Closthioamide is a potent inhibitor of bacterial DNA gyrase and highly active against Ec, MRSA, VRE and Mv), with MICs of 9.00 μM, 0.58 μM, 0.58 μM and 72.03 μM respectively.
Closthioamide is a potent inhibitor of bacterial DNA gyrase, isolated from the strictly anaerobic banterium Clostridium cellulolyticum and belongs to a new class of natural products[1]. In a standardized antimicrobial assay, it is found that closthioamide is highly active against a pathogenic, methicillin-resistant Staphylococcus aureus (MRSA) strain with a minimum inhibitory concentration (MIC) of 0.4 μg/mL-1 (0.58 μM). Closthioamide is even active against vancomycin-resistant Enterococcus faecalis (VRE) with the same low MIC value, and is thus significantly more potent against these bacteria than ciprofloxacin, the standard drugused against VRE, with remarkable strain selectivity. Furthermore, in a standardized cytotoxicity assay, closthioamide shows moderate antiproliferative and cytotoxic effects[2].
[1]. Chiriac AI, et al. Mode of action of closthioamide: the first member of the polythioamide class of bacterial DNA gyrase inhibitors. J AntimicrobChemother. 2015Sep;70(9):2576-88. [2]. Lincke T, et al. Closthioamide: an unprecedented polythioamide antibiotic from the strictly anaerobic bacterium Clostridium cellulolyticum.Angew Chem Int Ed Engl. 2010 Mar 8;49(11):2011-3.
| Cas No. | 1227367-59-0 | SDF | |
| Canonical SMILES | S=C(C1=CC=C(O)C=C1)NCCC(NCCC(NCCCNC(CCNC(CCNC(C2=CC=C(O)C=C2)=S)=S)=S)=S)=S | ||
| 分子式 | C29H38N6O2S6 | 分子量 | 695.04 |
| 溶解度 | Soluble in DMSO | 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 | 1.4388 mL | 7.1938 mL | 14.3877 mL |
| 5 mM | 0.2878 mL | 1.4388 mL | 2.8775 mL |
| 10 mM | 0.1439 mL | 0.7194 mL | 1.4388 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 |
| 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。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
In Vitro Susceptibility to Closthioamide among Clinical and Reference Strains of Neisseria gonorrhoeae
Antimicrob Agents Chemother 2017 Sep 22;61(10):e00929-17.PMID:28784667DOI:10.1128/AAC.00929-17.
Neisseria gonorrhoeae is one of the leading antimicrobial resistance threats worldwide. This study determined the MICs of Closthioamide to be 0.008 to 0.5 mg/liter for clinical N. gonorrhoeae strains and related species. Cross-resistance with existing antimicrobial resistance was not detected, indicating that Closthioamide could be used to treat drug-resistant N. gonorrhoeae.
An Unexpected Split-Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide
Angew Chem Int Ed Engl 2021 Feb 19;60(8):4104-4109.PMID:33119936DOI:10.1002/anie.202011741.
Closthioamide (CTA) is a symmetric nonribosomal peptide (NRP) comprised of two diaminopropane-linked polythioamidated monomers. CTA is biosynthesized by Ruminiclostridium cellulolyticum via an atypical NRP synthetase (NRPS)-independent biosynthetic pathway. Although the logic for monomer assembly was recently elucidated, the strategy for the biosynthesis and incorporation of the diamine linker remained a mystery. By means of genome editing, synthesis, and in vitro biochemical assays, we demonstrate that the final steps in CTA maturation proceed through a surprising split-merge pathway involving the dual use of a thiotemplated intermediate. This pathway includes the first examples of an aldo-keto reductase catalyzing the reductive release of a thiotemplated product, and of a transthioamidating transglutaminase. In addition to clarifying the remaining steps in CTA assembly, our data shed light on largely unexplored pathways for NRPS-independent peptide biosynthesis.
Reconstitution of Iterative Thioamidation in Closthioamide Biosynthesis Reveals Tailoring Strategy for Nonribosomal Peptide Backbones
Angew Chem Int Ed Engl 2019 Sep 9;58(37):13014-13018.PMID:31276268DOI:10.1002/anie.201905992.
Thioamide-containing nonribosomal peptides (NRPs) are exceedingly rare. Recently the biosynthetic gene cluster for the thioamidated NRP antibiotic Closthioamide (CTA) was reported, however, the enzyme responsible for and the timing of thioamide formation remained enigmatic. Here, genome editing, biochemical assays, and mutational studies are used to demonstrate that an Fe-S cluster containing member of the adenine nucleotide 伪-hydrolase protein superfamily (CtaC) is responsible for sulfur incorporation during CTA biosynthesis. However, unlike all previously characterized members, CtaC functions in a thiotemplated manner. In addition to prompting a revision of the CTA biosynthetic pathway, the reconstitution of CtaC provides the first example of a NRP thioamide synthetase. Finally, CtaC is used as a bioinformatic handle to demonstrate that thioamidated NRP biosynthetic gene clusters are more widespread than previously appreciated.
Mode of action of Closthioamide: the first member of the polythioamide class of bacterial DNA gyrase inhibitors
J Antimicrob Chemother 2015 Sep;70(9):2576-88.PMID:26174721DOI:10.1093/jac/dkv161.
Objectives: The spread of MDR bacteria represents a serious threat to human society and novel antibiotic drugs, preferably from new chemical classes, are urgently needed. Closthioamide was isolated from the strictly anaerobic bacterium Clostridium cellulolyticum and belongs to a new class of natural products, the polythioamides. Here, we investigated the antimicrobial activity and mechanism of action of Closthioamide. Methods: For assessing the antimicrobial activity of Closthioamide, MIC values and killing kinetics were determined. To identify its target pathway, whole-cell-based assays were used including analysis of macromolecular synthesis and recording the susceptibility profile of a library of clones with down-regulated potential target genes. Subsequently, the inhibitory effect of Closthioamide on the activity of isolated target enzymes, e.g. DNA gyrase and topoisomerase IV, was evaluated. Results: Closthioamide had broad-spectrum activity against Gram-positive bacteria. Notably, Closthioamide was very potent against MRSA and VRE strains. Closthioamide impaired DNA replication and inhibited DNA gyrase activity, in particular the ATPase function of gyrase and of topoisomerase IV, whereas there was little effect on the cleavage-rejoining function. Closthioamide also inhibited the relaxation activity of DNA gyrase, which does not require ATP hydrolysis, and thus may allosterically rather than directly interfere with the ATPase activity of gyrase. Cross-resistance to ciprofloxacin and novobiocin could not be detected in experimental mutants and clinical isolates. Conclusions: Closthioamide, a member of an unprecedented class of antibiotics, is a potent inhibitor of bacterial DNA gyrase; however, its molecular mechanism differs from that of the quinolones and aminocoumarins.
Mapping of the modular Closthioamide architecture reveals crucial motifs of polythioamide antibiotics
Chemistry 2014 Nov 17;20(47):15451-8.PMID:25284750DOI:10.1002/chem.201403836.
Closthioamide, the first known secondary metabolite from an anaerobic microorganism (Clostridium cellulolyticum), represents a highly potent antibiotic that is active against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE) at nanomolar concentrations. To unveil structure-activity relationships of the unusual polythioamide natural product we have designed a synthetic grid to access analogues with altered terminal aromatic moieties, diverse p-phenyl substituents, different types and sizes of aliphatic spacers, varying numbers of thioamide residues, and diverse sizes and symmetries of the poly-尾-thioalanyl backbone. A library of 28 Closthioamide analogues was tested against a panel of human pathogenic bacteria. We found that aromatic terminal groups, the defined length of the spacer groups, the presence of all six thioamide residues and the modular arrangement of the 尾-thioalanyl units play essential roles for the antibiotic activity of Closthioamide, yet there is a degree of freedom in the symmetry of the molecule. This study yields the first insights into pivotal structural motifs and the structural space of this new family of antibiotics, a prerequisite for the development of these promising antibiotics.