| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
|
| Targets |
PKA
|
|---|---|
| ln Vitro |
To investigate whether the stirnulatory action of H89 is mediated through an inhibitory action of PKA, we applied other types of PKA inhibitors and an inactive form of H89. H85, an inactive form of H89 (no inhibitory action on PKA), stimulated the amiloride-sensitive Isc (Fig. 3). H8, a PKA inhibitor, also stimulated the amiloride-sensitive Isc (Fig. 3). However, H8 required a higher concentration (100 j&l) to show the stimulatory action identical to that of 5 p.M H89, and 5 pM H8 had no significant action on the amiloride-sensitive Isc (Fig. 3). H7, a non-specific inhibitor of protein kinases including PKA and PKC, also required a high concentration, 100 pM, to show the PL-112 PKA-inhibition Independent Action of H89 Vol. 65, No. 10, 1999 stimulatory action on the amiloride-sensitive Isc. KT5720 (0.5 @VI), an inhibitor of PKA (14), did not stimulate the amiloride-sensitive Isc, but rather diminished it (Fig. 3). Further, another type of PICA inhibitor, Myr-PIU (3 @I) (15), also diminished the amiloride-sensitive Isc (Fig. 3). These observations suggest that all PKA inhibitors used in the present study do not have stimulatory action on the amiloride-sensitive Na+ transport, but that the stimulatory action is only seen in H-compounds. We also studied the effects of H8 and H7 on the stimulatory action of terbutaline on the amiloridesensitive Isc. Even in the presence of H8 or H7 of low concentration (5 pM) which did not stimulate the Isc, terbutaline still stimulated the amiloride-sensitive Isc; i.e., the stimulatory action of terbutaline was not blocked by H8 or H7 of a low concentration (5 pM). Although we need further experiments to confirm the signaling pathway of terbutaline and stimulatory mechanisms of H-compounds in regulation of the amiloride-sensitive Isc, we report here that H-compounds activate amiloride-sensitive Na+ channels resulting in stimulation of the amiloride-sensitive Na+ transport [1].
|
| Cell Assay |
Cell harvest and culture: [1]
Alveolar type II epithelial cells were isolated from the fetuses of pregnant Wistar rats (20 days’ gestational age; term = 22 days) which were completely anesthetized with inhalational ether (over dose) for 15 min. The epithelial cell was harvested from the fetuses and grown in primary culture according to the method previously described. In brief the lung fragments minced into 1 mm3 pieces obtained from fetuses were incubated at 37“C with 0.125% trypsin and 0.002% DNase and dissociated cells were then passed through a Nitex 100 mesh filter. The cells were then incubated with collagenase (0.1%) and purified by a differential adhesion technique. The majority of these cells are known to have morphologic and biochemical characteristics of alveolar type II epithelial cells. The cells were seeded at 3 x lo5 cells/well onto polycarbonate porous membranes in filter cups (Tissue culture-treated Transwell with 6.5 mm diameter) for Isc measurement or at 1 x lo6 cells/cm2 onto translucent porous Nunc filter inserts for single channel recording. All cells were grown in MEM with 10 % fetal bovine serum and penicillin-streptomycin at 37OC in a humidified 95 % air / 5 % CO:! environment. These epithelia were subsequently used 3 days after seeding under confluent conditions for short circuit current measurements or single channel recording. Cells plated on permeable supports formed polarized monolayers with their apical surfaces upward. Short circuit current measurement: [1] Monolayers were transferred to a modified Ussing chamber designed to hold the filter cup. Short-circuit currents were measured with an amplifier VCC-600. A positive current represents a net flow of cation from apical to basolateral solutions. Transepithelial voltage was measured with a pair of calomel electrodes which were immersed in a saturated KC1 solution and bridged to Ussing chamber by a pair of polyethylene tubes filled with a solution of 2% agarose in 2 M KCl. |
| References | |
| Additional Infomation |
The Na+ transport in alveolar type II epithelial cells of rat fetal lung was stimulated by cAMP, which is generally thought to act through activation of protein kinase A (PKA). PKA inhibitors (H8, H89 and H7) stimulated amiloride-sensitive Na+ transport in the alveolar type II epithelial cells. H85, an inactive form of H89 as a PKA inhibitor, had also mimicked the stimulatory action of H89 on the Na+ transport. On the other hand, another type of PKA inhibitor, KT5720 or myristoylated PKA inhibitory peptide [14-22] amide, did not stimulate the Na+ transport, but inhibited the Na+ transport unlike H-compounds. These observations suggest that H-compounds act on the Na+ transport depending on the structure.[1]
Taken together, these observations indicate that the stimulatory action of H-compounds on the __. . amiloride-sensitive Na+ transport is based upon their structures. Figure 4 shows the structure ot compounds used in the present study. The H-compounds (H89, H85, H8 and H7) contains 5- isoquinolinesulfone. Other PKA inhibitors, KT5720 and Myr-PKI, have completely different structures (Fig. 4). Although the power (sensitivity) of the H-compound in stimulation of the amiloride-sensitive Na+ transport depends upon the structure of side chain, the structure of 5- isoquinolinesulfone is strongly suggested to play a role in stimulation of the Na+ transport. |
| Molecular Formula |
C12H17CL2N3O2S
|
|---|---|
| Molecular Weight |
338.24
|
| Exact Mass |
337.042
|
| Elemental Analysis |
C, 42.61; H, 5.07; Cl, 20.96; N, 12.42; O, 9.46; S, 9.48
|
| CAS # |
113276-94-1
|
| Related CAS # |
84478-11-5
|
| PubChem CID |
150584
|
| Appearance |
White to off-white solid powder
|
| Boiling Point |
473.9ºC at 760 mmHg
|
| Melting Point |
221-222℃ (methanol water )
|
| Flash Point |
240.4ºC
|
| Vapour Pressure |
3.79E-09mmHg at 25°C
|
| LogP |
4.199
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
20
|
| Complexity |
353
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CNCCNS(=O)(=O)C1=CC=CC2=C1C=CN=C2.Cl.Cl
|
| InChi Key |
RJJLZYZEVNCZIW-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C12H15N3O2S.2ClH/c1-13-7-8-15-18(16,17)12-4-2-3-10-9-14-6-5-11(10)12;;/h2-6,9,13,15H,7-8H2,1H3;2*1H
|
| Chemical Name |
N-(2-(methylamino)ethyl)isoquinoline-5-sulfonamide dihydrochloride
|
| Synonyms |
H8 dihydrochloride; H-8 dihydrochloride; 113276-94-1; H-8, Dihydrochloride; H-8 (dihydrochloride); N-[2-(Methylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride; H8 dihydrochloride; N-(2-(Methylamino)ethyl)isoquinoline-5-sulfonamide dihydrochloride; H8; H8
|
| 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, avoid exposure to moisture. |
| 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 : ~20.83 mg/mL (~61.58 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.39 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to 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 (7.39 mM) (saturation unknown) 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. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.39 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9565 mL | 14.7824 mL | 29.5648 mL | |
| 5 mM | 0.5913 mL | 2.9565 mL | 5.9130 mL | |
| 10 mM | 0.2956 mL | 1.4782 mL | 2.9565 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
