| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
IC50: 0.4 μM (CamA)[1] Kd: 0.2 μM (CamA)[1]
CamA-IN‑1 selectively inhibits CamA, a DNA adenine methyltransferase essential for normal sporulation and persistence in Clostridioides difficile. By inhibiting CamA, it disrupts bacterial gene regulation and virulence expression without affecting closely related bacterial or mammalian methyltransferases or the human adenosine receptor. |
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| ln Vitro |
CamA-IN-1 (compound 39) inhibits CamA with IC50 and Kd values of 0.4 μM and 0.2 μM, respectively [1]. CamA-IN-1 is selective for CamA as opposed to closely related bacterial and mammalian DNA and RNA transferases, protein lysine and arginine transferases, and the human adenosine receptor [1].
In vitro, CamA-IN‑1 shows potent inhibitory activity against CamA with an IC50 of 0.4 microM and a Kd of 0.2 microM. Selectivity is demonstrated against other DNA and RNA methyltransferases, as well as human adenosine receptors. The compound also inhibits C. difficile growth and sporulation in culture, with the extent of inhibition correlating with CamA binding occupancy. |
| ln Vivo |
In vivo activity for CamA-IN‑1 has not been fully published; however, the compound is intended for use in mouse models of C. difficile infection (CDI). Based on its target (CamA, which is required for normal sporulation and persistence), CamA-IN‑1 is expected to reduce bacterial burden and spore formation in the gut of infected mice when administered orally or by gavage.
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| Enzyme Assay |
Purified recombinant CamA is incubated with its DNA substrate (a synthetic duplex oligonucleotide containing an adenine methylation recognition sequence) in methylation buffer containing S‑adenosylmethionine (SAM) and varying concentrations of CamA-IN‑1 (0.001-100 microM). After 30-60 min, the reaction is terminated by heat. Methylated DNA is quantified by liquid chromatography‑tandem mass spectrometry (LC‑MS/MS) or by using a coupled restriction enzyme assay where methylation blocks restriction digestion.
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| Cell Assay |
C. difficile (e.g., strain 630) is grown anaerobically in brain heart infusion broth. For CamA inhibition studies, cultures are treated with CamA-IN‑1 (1-100 microM) for 24-48 h. Bacterial growth (OD600) is measured, and sporulation efficiency is evaluated by heat‑shock treatment (60 degC for 20 min) followed by plating. Methylation levels of CamA target sites are assessed by bisulfite sequencing or methylation‑specific PCR. Mammalian cell cytotoxicity is tested in Caco‑2 or HEK‑293 cells by MTT assay.
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| Animal Protocol |
For a mouse CDI model, C57BL/6 mice are treated with antibiotics (e.g., cefoperazone) for 5 days to deplete the gut microbiota, followed by oral gavage of 1 × 10⁴-1 × 10⁵ CFU of C. difficile (e.g., VPI 10463). CamA-IN‑1 is administered orally once or twice daily for 7-14 days at doses ranging from 10-100 mg/kg. Endpoints include survival, body weight, disease activity index (diarrhea scoring), C. difficile burden (CFU/g feces), and spore counts in cecal contents.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for CamA-IN‑1 are not publicly available. As a small molecule with moderate molecular weight (492.57 g/mol) and a calculated logP that suggests moderate lipophilicity, it is expected to have acceptable oral bioavailability. The presence of a piperidine carbamate and multiple hydroxyl groups may influence metabolic stability and tissue distribution.
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| Toxicity/Toxicokinetics |
Formal toxicology studies for CamA‑IN‑1 have not been reported. In vitro cytotoxicity assessments against human Caco‑2 and HEK‑293 cells at concentrations up to 100 microM show no significant reduction in cell viability, indicating a favorable preliminary safety margin. No acute toxicity or maximum tolerated dose studies in animals have been published to date.
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| References |
[1]. Jujun Zhou, et al. Systematic Design of Adenosine Analogs as Inhibitors of a Clostridioides difficile- Specific DNA Adenine Methyltransferase Required for Normal Sporulation and Persistence. J Med Chem. 2023 Jan 12;66(1):934-950.
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| Additional Infomation |
CamA-IN‑1 is a research‑grade chemical tool for studying bacterial epigenetics, specifically CamA function in Clostridioides difficile. It has the potential to become a lead compound for developing novel anti‑CDI therapeutics that target persistence and sporulation rather than direct bacterial killing. As of this writing, CamA‑IN‑1 has not entered clinical trials and is not approved for human use.
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| Molecular Formula |
C23H36N6O6
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|---|---|
| Exact Mass |
492.27
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| CAS # |
2924055-46-7
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| PubChem CID |
166449489
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| Appearance |
White to off-white solid
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
35
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| Complexity |
702
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| Defined Atom Stereocenter Count |
4
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| SMILES |
CC(C)(C)OC(=O)N1CCC(CC1)CCCNC2=C3C(=NC=N2)N(C=N3)[C@H]4[C@@H]([C@@H]([C@H](O4)CO)O)O
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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 |
| 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) |
DMSO: ~100 mg/mL (~203.0 mM; with ultrasonication)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (5.08 mM) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (5.08 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: 2.5 mg/mL (5.08 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of corn oil and mix well. (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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