| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| Other Sizes |
|
| Targets |
Acetylcholinesterase (AChE)
|
|---|---|
| ln Vitro |
Nerve agents (NA) are chemical warfare munitions and their exposure causes a progressive inhibition of acetylcholinesterase (AChE). This inhibition causes NA-induced brain damage in central nervous system (CNS). Oximes reactivate AChE in both the peripheral nervous system and the CNS. Transport of the oxime across the blood-brain barrier (BBB) in the existed therapeutic concentrations at the brain parenchyma determines the effectiveness of antidote therapy on respiratory depression and NA-induced brain damage. However, oximes could not cross the BBB in therapeutic concentrations. The aim of this study was to load AChE reactivator obidoxime chloride to PLGA and PEG-b-PLGA nanoparticles and to improve the BBB transport of the molecule. Brain microvascular endothelial cells were used as the BBB model. 79.3 ± 4.2% of obidoxime was released from PLGA nanoparticles and 88.2 ± 4.4% of obidoxime was released from PEG-b-PLGA nanoparticles within 24 h. It was found that PEG-b-PLGA nanoparticles were ideal drug carrier because of its low tissue toxicity, few side effects, and controllable drug release profile. Transport efficiency of obidoxime across the BBB is a major challenge in the prevention of the CNS, the effectiveness of NA poisoning and new strategies like using obidoxime-loaded PEG-b-PLGA nanoparticles could overcome this challenge for the management of NA-induced brain damage[2].
|
| ln Vivo |
Orbidoxime dichloride has been tested to lessen the acute toxicity of sarin and has an LD50 of 57.7 mg/kg following sarin exposure (intramuscular injection; 10 mg/kg; after 60 minutes) [1].
|
| Enzyme Assay |
In vitro drug release studies
For determining the amount of obidoxime released from the nanoparticles in vitro, a dynamic dialysis bag diffusion technique was used. PLGA and PEG-b-PLGA nanoparticles which contained 0.3 mg obidoxime, were placed in the dialysis bag (diameter 1 cm, molecular weight cut-off 12−14 kDa) which was containing 1 mL of isotonic phosphate buffered saline (pH 7.4), respectively. The dialysis bag which was filled with the nanosuspension was immersed into 30 mL isotonic phosphate buffered saline (pH 7.4) and then it was incubated in a water-bath shaker at 37 °C with constant orbital mixing at 30 rpm. At specified predetermined time intervals (1, 2, 3, 4, 5, 6, 8, 12, 24 h), an aliquot of the release medium (1 mL) was removed from the receptor compound and the removed liquid was replaced with fresh medium. The obidoxime concentrations of the aliquots were determined by using HPLC as described above, and the cumulative amount of obidoxime which released from the nanoparticles was calculated. The cumulative percentage of released obidoxime was plotted versus time. Each data point (SD) is the average of three consecutive[1].
|
| Cell Assay |
Colorimetric cell viability via MTT assay[1]
The cell viability of BMVEC lines after adding obidoxime solution, obidoxime-loaded PLGA and PEG-b-PLGA nanoparticles was measured by MTT (Danhier et al., 2009). For the assay, the cells were seeded into 96-well plates at the density of 3 × 104 cells/cm2. Cells were incubated until they were approximately 70–80% confluent. Obidoxime solutions (10 mM, 2.5 mM, 1 mM, 0.25 mM and 0.1 mM) and nanoparticles containing 0.1−10 mM obidoxime were added to the cells in 96-well plates and they were incubated for 24, 48, and 72 h. After the incubation, 20 μL of the medium containing MTT was added to each well and the plates were shaken for 60 s. The absorbance of each plate was measured by using a microplate reader at 550 and 690 nm. The medium was also used as a positive control. The untreated controls were also measured for the determination of 100% cell viability in each plate. Cell-monolayer permeation studies[1] For the BBB transport experiments, BMVEC were plated on transwell filters (polycarbonate 12 well, pore size 0.4 μm; Millipore) in a density of 3 × 104 cells/cm2. Twenty-four hour before each transport experiment, the medium was replaced with the fresh one. For checking the existence of junctions and confluent monolayer, the cell monolayer was periodically inspected under a microscope and the transendothelial electrical resistance (TEER) was measured with EVOMX Epithelial Volt-Ohm meter, World Precision Instruments. The transport was estimated by placing obidoxime solution or obidoxime-loaded PLGA and PEG-b-PLGA nanoparticles on the upper side of the monolayers and measuring obidoxime in the well media at various time intervals (30, 60, 120, 180, and 240 min). The obidoxime concentrations of the aliquots were determined by HPLC as described above, and the cumulative amount of obidoxime permeated from the cell monolayer was calculated. |
| Animal Protocol |
Animal/Disease Models: NMRI male mice
Doses: 10 mg/kg Route of Administration: intramuscularinjection Experimental Results: It can effectively reactivate sarin-inhibited acetylcholinesterase (AChE). |
| References |
[1]. Kassa J,et al. A comparison of the reactivating and therapeutic efficacy of two novel bispyridinium oximes (K727, K733) with the oxime HI-6 and obidoxime in sarin-poisoned rats and mice.Toxicol Mech Methods. 2015 Mar;25(3):229-33.
[2]. Toxicol Lett . 2020 May 5:330:53-58. doi: 10.1016/j.toxlet.2020.03.018. |
| Additional Infomation |
Cholinesterase reactivator occurring in two interchangeable isomeric forms, syn and anti.
See also: Obidoxime (annotation moved to). |
| Molecular Formula |
C14H16N4O3CL2
|
|---|---|
| Molecular Weight |
359.20784
|
| Exact Mass |
358.06
|
| CAS # |
114-90-9
|
| Related CAS # |
2337-82-8 (Br);73840-43-4 (sulfate);4605-73-6 (iodide);114-90-9 (Cl);
|
| PubChem CID |
135566074
|
| Appearance |
Off-white to light yellow solid powder
|
| Density |
1.33g/cm3
|
| Boiling Point |
1132.2ºC at 760 mmHg
|
| Melting Point |
216-219ºC dec.
|
| Flash Point |
638.5ºC
|
| Vapour Pressure |
0mmHg at 25°C
|
| Index of Refraction |
1.606
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
23
|
| Complexity |
303
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CN(C=CC1=CN=O)COCN2C=CC(=CN=O)C=C2.Cl.Cl
|
| InChi Key |
ZIFJVJZWVSPZLE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C14H14N4O3.2ClH/c19-15-9-13-1-5-17(6-2-13)11-21-12-18-7-3-14(4-8-18)10-16-20;;/h1-10H,11-12H2;2*1H
|
| Chemical Name |
(NE)-N-[[1-[[4-[(E)-hydroxyiminomethyl]pyridin-1-ium-1-yl]methoxymethyl]pyridin-1-ium-4-yl]methylidene]hydroxylamine;dichloride
|
| Synonyms |
OBIDOXIME CHLORIDE; Obidoxime; Obidoxime dichloride; Obidoxim; 114-90-9; Obidoxime (dichloride); 3HXR312Z9M; Toksobidin;
|
| 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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 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 | 2.7839 mL | 13.9194 mL | 27.8389 mL | |
| 5 mM | 0.5568 mL | 2.7839 mL | 5.5678 mL | |
| 10 mM | 0.2784 mL | 1.3919 mL | 2.7839 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
