| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg | |||
| Other Sizes |
| Targets |
The primary molecular target of netropsin is DNA, specifically the minor groove of double-stranded DNA. Netropsin binds preferentially to AT-rich regions of the minor groove, interacting with the DNA bases through hydrogen bonding and van der Waals interactions. The compound binds to DNA with high affinity, with a binding constant (Ka) of 4.9 × 10⁵ M⁻¹ or higher. By binding to the minor groove, netropsin interferes with DNA replication and transcription processes. The compound also attenuates NOS2 (inducible nitric oxide synthase) induction by interfering with HMGA1 DNA binding to the core NOS2 promoter, thereby reducing NO production and improving survival in endotoxemia. Netropsin's binding to DNA also provides radioprotective ability against radiation-induced damage.
|
|---|---|
| ln Vitro |
Netropsin diHClide binds to dsDNA non-insertively, which may confer antibiotic and antiviral effects. It also attenuates NOS2 induction by obstructing HMGA1 DNA's binding to the core NOS2 promoter, which increases the survival rate of endotoxemia in mice [2]. Because of its strong binding affinity and great structural stability, neraptin dihydrochloride in DNA solution can protect DNA from radiation damage. The Netropsin-DNA complex is not cleaved radiatively by the hydroxyl radical (OH*) produced by ionizing radiation [3].
In vitro studies demonstrate that netropsin binds preferentially to AT-rich regions of the minor groove of DNA with high affinity (Ka = 4.9 × 10⁵ M⁻¹). The compound binds to double-stranded DNA in a non-intercalative manner. Netropsin has potential antibiotic and antiviral properties by binding to viral DNA and interfering with replication and transcription. The compound attenuates NOS2 induction by interfering with HMGA1 DNA binding to the core NOS2 promoter. In DNA solutions, netropsin has radioprotective ability against radiation-induced damage due to its high binding affinity and structural stability. Netropsin has been ranked #1 in DNA binding affinity among five common minor groove binders, ensuring reliable AT-rich sequence targeting for DNase I footprinting. These in vitro activities make netropsin a powerful tool in biochemical and pharmacological investigations. |
| ln Vivo |
In vivo studies of netropsin dihydrochloride have demonstrated its therapeutic potential. The compound improves survival from murine endotoxaemia by attenuating NOS2 induction through interference with HMGA1 DNA binding to the core NOS2 promoter. In mouse models of endotoxemia, netropsin treatment increases survival rates. The compound's antibiotic and antiviral properties have also been studied in animal models of infection. Netropsin binds to AT-rich regions of viral DNA, providing a tool to investigate replication and transcription inhibition mechanisms. In vivo protocols typically involve intraperitoneal or intravenous administration of netropsin at doses determined from preliminary toxicity studies. Endpoints include survival, bacterial load, cytokine levels, and tissue histopathology. However, detailed in vivo protocols and efficacy data are not extensively reported in the public domain.
|
| Enzyme Assay |
For DNA binding assays, netropsin binding to DNA is measured using various biophysical techniques. For UV-Vis or fluorescence spectroscopy, netropsin is dissolved in buffer (e.g., 10 mM Tris-HCl, pH 7.4, 100 mM NaCl) and titrated with increasing concentrations of calf thymus DNA or synthetic AT-rich oligonucleotides. Absorbance or fluorescence changes (quenching or enhancement) are monitored, and binding parameters (Ka, binding stoichiometry) are calculated from binding isotherms. For circular dichroism (CD) spectroscopy, the CD spectrum of netropsin is measured in the presence and absence of DNA. For DNase I footprinting, a DNA fragment containing a known AT-rich sequence is labeled with ³²P at one end, incubated with netropsin (0.1-100 µM), and then digested with DNase I. The digestion products are separated by denaturing polyacrylamide gel electrophoresis, and the protected regions (footprints) are identified by autoradiography. For NOS2 promoter binding assays, the ability of netropsin to interfere with HMGA1 DNA binding to the core NOS2 promoter is assessed by electrophoretic mobility shift assay (EMSA) or chromatin immunoprecipitation (ChIP).
|
| Cell Assay |
For cellular studies, cells (e.g., macrophages, cancer cells, or other cell lines) are cultured in appropriate medium (RPMI-1640 or DMEM) with 10% FBS and antibiotics. Cells are seeded in 6-well or 96-well plates. Netropsin dihydrochloride is dissolved in sterile water or PBS and diluted in culture medium to final concentrations (typically 0.1-100 µM). For NOS2 induction studies, cells (e.g., RAW 264.7 macrophages) are treated with netropsin for 1-2 hours, then stimulated with LPS (0.1-1 µg/ml) and/or IFN-γ for 4-24 hours. NOS2 expression is assessed by Western blot, and NO production is measured by Griess assay. For DNA damage studies, cells are treated with netropsin and DNA damage is assessed by comet assay or γ-H2AX staining. For radioprotection studies, cells are treated with netropsin before exposure to ionizing radiation, and cell viability and DNA damage are assessed. For antiviral studies, virus-infected cells are treated with netropsin, and viral replication is measured by plaque assay or qPCR.
|
| Animal Protocol |
For in vivo studies, adult mice (6-8 weeks old) are used. For endotoxemia models, mice are injected intraperitoneally with LPS (10-20 mg/kg) to induce endotoxemia. Netropsin dihydrochloride is dissolved in sterile saline and administered intraperitoneally or intravenously at doses of 1-50 mg/kg, typically as a single dose or multiple doses before or after LPS challenge. Survival is monitored for 24-72 hours. Blood samples are collected for measurement of NO, cytokines (TNF-α, IL-1β, IL-6), and liver enzymes. Liver and lung tissues are collected for histopathological examination. For antiviral studies, mice are infected with virus, and netropsin is administered prophylactically or therapeutically. Viral load in tissues is measured by plaque assay or qPCR. For radioprotection studies, mice are treated with netropsin before whole-body irradiation, and survival, hematopoietic recovery, and DNA damage are assessed.
|
| ADME/Pharmacokinetics |
Pharmacokinetic data for netropsin dihydrochloride are limited, as the compound is used as a research reagent rather than a therapeutic agent. As a small molecule with DNA-binding properties, netropsin is expected to have distribution to various tissues, particularly those with high cell turnover. The compound's metabolism likely involves hepatic metabolism, and it is cleared via biliary and renal excretion. However, no dedicated ADME studies have been reported for netropsin. The compound is primarily used in vitro and is not intended for in vivo pharmacokinetic characterization.
|
| Toxicity/Toxicokinetics |
Toxicological data for netropsin are limited, as the compound is used as a research reagent. The compound is a basic cytotoxic polypeptide and may have cytotoxicity at higher concentrations. In cellular assays, netropsin shows concentration-dependent effects on cell viability. No acute toxicity LD50 values or organ-specific toxicity data have been reported. As with all chemical reagents, appropriate safety precautions should be taken when handling netropsin dihydrochloride, including the use of personal protective equipment (gloves, lab coat, safety glasses) and work in a well-ventilated area.
|
| References |
|
| Additional Infomation |
A basic polypeptide isolated from Streptomyces netropsis. It is cytotoxic and can bind strongly and specifically to the AT region of DNA, thus making it suitable for genetic research.
Netropsin dihydrochloride (CAS 18133-22-7) is a DNA minor groove binder originally isolated from Streptomyces netropsis. It binds preferentially to AT-rich regions of the minor groove of DNA with high affinity (Ka = 4.9 × 10⁵ M⁻¹), interfering with replication and transcription processes. The compound has potential antibiotic and antiviral properties and improves survival from murine endotoxaemia by attenuating NOS2 induction. Netropsin has radioprotective ability against radiation-induced damage. It is ranked #1 in DNA binding affinity among minor groove binders and is widely used as a molecular probe in nucleic acid research. Netropsin dihydrochloride is strictly for research use only. |
| Molecular Formula |
C18H26N10O3.HCL
|
|---|---|
| Molecular Weight |
466.92518
|
| Exact Mass |
466.196
|
| CAS # |
18133-22-7
|
| Related CAS # |
1438-30-8;18133-22-7 (HCl);63770-20-7 (Alt. HCl);
|
| PubChem CID |
5351456
|
| Appearance |
Off-white to yellow solid powder
|
| LogP |
2.423
|
| Hydrogen Bond Donor Count |
8
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
32
|
| Complexity |
723
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
FSOYOTWCAMRRGU-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C18H26N10O3.ClH/c1-27-9-11(6-12(27)16(30)23-4-3-14(19)20)26-17(31)13-5-10(8-28(13)2)25-15(29)7-24-18(21)22;/h5-6,8-9H,3-4,7H2,1-2H3,(H3,19,20)(H,23,30)(H,25,29)(H,26,31)(H4,21,22,24);1H
|
| Chemical Name |
N-[5-[(3-amino-3-iminopropyl)carbamoyl]-1-methylpyrrol-3-yl]-4-[[2-(diaminomethylideneamino)acetyl]amino]-1-methylpyrrole-2-carboxamide;hydrochloride
|
| HS Tariff Code |
2934.99.9001
|
| 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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~248.32 mM)
H2O : ~20 mg/mL (~39.73 mM) |
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1416 mL | 10.7082 mL | 21.4165 mL | |
| 5 mM | 0.4283 mL | 2.1416 mL | 4.2833 mL | |
| 10 mM | 0.2142 mL | 1.0708 mL | 2.1416 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
