| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
GABA-C receptor
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|---|---|
| ln Vitro |
The potency of a series of eight compounds structurally related with 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP), a potent GABA(A) partial agonist exhibiting GABA(C) rho(1) antagonist effect (K(i)=25 microM), was determined electrophysiologically using homomeric human GABA(C) rho(1) receptors expressed in Xenopus oocytes. Protolytic properties (pK(a) values for the acidic bioisosteric groups) and the presence of steric bulk in the molecules appear to be structural parameters of importance for blockade of the GABA(C) rho(1) receptor. Within this series of moderately potent GABA(C) antagonists, only 4,5,6,7-tetrahydropyrazolo[5,4-c]pyridin-3-ol (Aza-THIP) does not interact detectably with GABA(A) receptors, and Aza-THIP has the potential of being a useful tool for molecular and behavioural pharmacological studies. [1]
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| Enzyme Assay |
Electrophysiological recording [1]
Xenopus laevis was anaesthesized with 0.17% ethyl 3-aminobenzoate and a lobe of the ovaries was carefully removed. The lobe of the ovary was placed in oocyte releasing buffer 2 (OR2) (82.5 mM NaCl, 2 mM KCl, 1 mM MgCl2 · 6H2O, 5 mM HEPES, pH 7.5) with 2 mg/mL Collagenase A for 2 h. Defolliculated oocytes were then rinsed with frog Ringer solution (96 mM NaCl, 2 mM KCl, 1 mM MgCl2 · 6H2O, 1.8 mM CaCl2, 5 mM HEPES, pH 7.5) supplemented with 2.5 mM pyruvate, 0.5 mM theophylline and 50 μg/mL gentamycin. Stage V–VI oocytes were collected. Receptor activity was measured by two electrode voltage clamp recording using a Geneclamp 500 amplifier, a MacLab 2e recorder and Chart version 3.5 program. Oocytes were voltage clamped at −60 mV and continuously superfused with frog Ringer solution (96 mM NaCl, 2 mM KCl, 1 mM MgCl2 · 2H2O, 1.8 mM CaCl2 · 2H2O, 5 mM HEPES). For receptor activation measurements, the indicated concentrations of drug were added to the buffer solution. Antagonist activity was measured as a pKi or an IC50 value. The pKi value was determined on the basis of a GABA concentration-response curve as a control followed by a GABA concentration-response curve in the presence of a fixed antagonist concentration on the same oocyte (n≥3). The IC50 value was determined by measuring the activation of the receptor by GABA in the presence of different antagonist concentrations. The GABA concentration was chosen as the concentration which produces 50% of max response, e.g., EC50, based on a standard GABA concentration–response curve for each oocyte prior to the inhibition curve, thereby using a fixed relative response level of GABA. |
| References |
[1]. Aza-THIP and related analogues of THIP as GABA C antagonists. Bioorg Med Chem. 2003 Nov 17;11(23):4891-6.
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| Additional Infomation |
To date, TPMPA has been the most useful antagonist for studying GABAC receptors. However, Aza-THIP has been shown to be a specific competitive GABAC antagonist, serving as a complement to TPMPA and a reference compound for GABAC receptors. TPMPA exhibits potent antagonistic activity against GABAC receptors, while Aza-THIP shows weaker activity against both GABAA receptors (Table 1) and metabolized GABAB receptors. Using a series of THIP-related compounds, we found that only minor structural changes affect their potency as GABAC antagonists. The 3-isoxazole ring of Iso-THIP (pKa 3.0) is more acidic than that of THIP (pKa 4.4) (Table 1), suggesting that the difference in potency may be due to a stronger electrostatic interaction between Iso-THIP and the receptor. The structural and protonation similarities between THIP and Aza-THIP indicate that the latter, like THIP, can cross the blood-brain barrier. Therefore, although Aza-THIP has only moderate potency as a GABAC antagonist, it may be a useful compound for behavioral pharmacology studies. [1]
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| Molecular Weight |
139.15516
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|---|---|
| Exact Mass |
139.075
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| Elemental Analysis |
C, 51.79; H, 6.52; N, 30.20; O, 11.50
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| CAS # |
654666-65-6
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| PubChem CID |
55285412
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| Appearance |
Typically exists as solid at room temperature
|
| LogP |
0.089
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
10
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| Complexity |
209
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| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1CNCC2=C1C(=O)NN2
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| InChi Key |
UPMMUUFLHUEUKP-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C6H9N3O/c10-6-4-1-2-7-3-5(4)8-9-6/h7H,1-3H2,(H2,8,9,10)
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| Chemical Name |
1,2,4,5,6,7-hexahydropyrazolo[3,4-c]pyridin-3-one
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| Synonyms |
aza-THIP; 654666-65-6; 1H,4H,5H,6H,7H-pyrazolo[3,4-c]pyridin-3-ol; 1H,4H,5H,6H,7H-pyrazolo(3,4-c)pyridin-3-ol; 1,2,4,5,6,7-Hexahydro-3H-pyrazolo[3,4-c]pyridin-3-one; 4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridin-3-ol; 4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-ol; 4,5,6,7-tetrahydropyrazolo[5,4-c]pyridin-3-ol;
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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) |
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
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|---|---|
| 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 | 7.1860 mL | 35.9299 mL | 71.8597 mL | |
| 5 mM | 1.4372 mL | 7.1860 mL | 14.3719 mL | |
| 10 mM | 0.7186 mL | 3.5930 mL | 7.1860 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.
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