| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
cyclic GMP-dependent protein kinase, cyclic AMP-dependent protein[2]
HA-1004 targets multiple kinases including PKG, PKA, MLCK, and PKC. Protein kinase G (PKG) and protein kinase A (PKA) are key mediators of cyclic nucleotide signaling, regulating smooth muscle tone, platelet function, and cardiac contractility. Myosin light-chain kinase (MLCK) phosphorylates myosin light chain, playing a critical role in smooth muscle contraction. Protein kinase C (PKC) is involved in various cellular processes including cell growth, differentiation, and apoptosis. By inhibiting these kinases, HA-1004 modulates multiple signaling pathways, leading to vasodilation and inhibition of lipolysis. The compound's multi-target profile makes it useful for studying the roles of these kinases in various physiological and pathological processes. |
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| ln Vitro |
In vitro studies have demonstrated that HA-1004 is a selective inhibitor of PKG and PKA with Kis of 1.3 and 2.3 μM, respectively. The compound is less potent against MLCK (Ki = 70 μM) and PKC (Ki = 18 μM). HA-1004 induces relaxation of isolated rabbit aortic strips precontracted with histamine, demonstrating its vasodilatory activity. The compound inhibits lipolysis in adipocytes through inhibition of PKA-mediated signaling. HA-1004 has been used to study the role of cyclic nucleotide-dependent protein kinases in smooth muscle relaxation, platelet aggregation, and other cellular processes. The compound's activity is concentration-dependent and reversible.
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| ln Vivo |
Simultaneously acting with morphine, HA-1004 (40 nmol/day; administered by micropump, flow rate = 1 μL/h) phosphorylates ERK1/2 and TH during morphine withdrawal in rats, preventing the rise in NA conversion [ 1].
In vivo studies with HA-1004 have demonstrated its vasodilatory effects. The compound induces vascular relaxation in isolated tissue preparations such as rabbit aortic strips. While comprehensive in vivo data in animal models is limited in the available literature, HA-1004 has been used as a pharmacological tool to study the roles of PKA and PKG in cardiovascular function. The compound's ability to inhibit cyclic nucleotide-dependent protein kinases makes it useful for investigating the physiological roles of these enzymes in blood pressure regulation, platelet function, and other cardiovascular processes. |
| Enzyme Assay |
Cell-free kinase assays for HA-1004 typically measure inhibition of kinase activity using radiolabeled ATP or fluorescence-based methods. A standard protocol for PKA inhibition involves incubating the catalytic subunit of PKA with varying concentrations of HA-1004 (0.1-100 μM), a peptide substrate (e.g., Kemptide), and [γ-³²P]ATP in kinase buffer at 30°C for 10-30 minutes. The reaction is terminated by spotting onto P81 phosphocellulose paper, washing with phosphoric acid, and counting incorporated radioactivity. For PKG assays, a similar protocol is used with the appropriate kinase and substrate. IC₅₀ or Ki values are determined from dose-response curves using nonlinear regression analysis.
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| Cell Assay |
Cell-based assays for HA-1004 typically use smooth muscle cells or other cell types to assess its effects on kinase signaling and cellular function. A standard protocol involves culturing vascular smooth muscle cells in 96-well plates, treating with HA-1004 at concentrations ranging from 0.1-100 μM for 1-24 hours, and measuring downstream effects. For assessing vasodilation, isolated aortic rings are mounted in tissue baths, precontracted with histamine or phenylephrine, and relaxation is measured after HA-1004 addition. Phosphorylation of kinase substrates is assessed by Western blotting using phospho-specific antibodies. Cell viability and proliferation can be assessed by MTT or other assays.
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| Animal Protocol |
In vivo animal studies with HA-1004 are primarily conducted using isolated tissue preparations rather than whole animal models. For vascular relaxation studies, a standard protocol involves euthanizing rabbits, removing the thoracic aorta, cutting into rings, and mounting them in organ baths containing physiological salt solution. The rings are precontracted with histamine, and cumulative concentrations of HA-1004 (1-100 μM) are added to measure relaxation. For systemic administration studies, HA-1004 could be administered intravenously or intraperitoneally to rodents, with monitoring of blood pressure, heart rate, and other cardiovascular parameters.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for HA-1004 is limited in the published literature, as the compound is primarily used as a research tool rather than a drug candidate. The compound's molecular weight of 293.34 g/mol and its physicochemical properties suggest reasonable cell permeability. For studies requiring systemic administration, PK parameters would need to be determined empirically. The compound is typically dissolved in DMSO for in vitro studies and may require appropriate formulation for in vivo administration. Metabolism and clearance pathways have not been extensively characterized.
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| Toxicity/Toxicokinetics |
Toxicological data specific to HA-1004 is limited, as the compound is primarily used in research settings. At effective concentrations for kinase inhibition (low micromolar range), the compound does not show significant cytotoxicity in most cell types. Higher concentrations may have off-target effects or cytotoxicity. As with all kinase inhibitors, potential toxicity could arise from inhibition of multiple kinases, and the compound's multi-target profile may contribute to broader biological effects. Standard laboratory safety precautions should be observed when handling this compound.
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| References |
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| Additional Infomation |
2-[2-(5-isoquinolinylsulfonylamino)ethyl]guanidine is a member of the isoquinoline class of compounds.
HA-1004 is a research compound and not an approved drug. No clinical trials or regulatory approvals exist for this compound. It is commercially available from various suppliers for research use only. The compound's primary value lies in its utility as a pharmacological tool for studying cyclic nucleotide-dependent protein kinases (PKA and PKG) and other kinases (MLCK and PKC). By inhibiting these kinases with varying potencies, HA-1004 enables researchers to dissect the roles of these enzymes in smooth muscle physiology, platelet function, and other biological processes. It is particularly useful for studying the mechanisms of vasodilation and the regulation of smooth muscle tone. |
| Molecular Formula |
C12H15N5O2S
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|---|---|
| Molecular Weight |
293.34
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| Exact Mass |
293.095
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| CAS # |
91742-10-8
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| Related CAS # |
HA-1004 hydrochloride; 92564-34-6; HA-1004 dihydrochloride; 92564-08-4
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| PubChem CID |
3546
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| Appearance |
White to off-white solid powder
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| LogP |
2.658
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
20
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| Complexity |
441
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC2=C(C=CN=C2)C(=C1)S(=O)(=O)NCCN=C(N)N
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| InChi Key |
MZNDNBFMSVMUCX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C12H15N5O2S/c13-12(14)16-6-7-17-20(18,19)11-3-1-2-9-8-15-5-4-10(9)11/h1-5,8,17H,6-7H2,(H4,13,14,16)
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| Chemical Name |
2-[2-(isoquinolin-5-ylsulfonylamino)ethyl]guanidine
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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: ~50 mg/mL (~170.5 mM; with ultrasonication)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (4.26 mM) (saturation unknown) 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.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (12.5 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: ≥ 1.25 mg/mL (4.26 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 DMSO stock solution (12.5 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: ≥ 1.25 mg/mL (4.26 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 DMSO stock solution (12.5 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.4090 mL | 17.0451 mL | 34.0901 mL | |
| 5 mM | 0.6818 mL | 3.4090 mL | 6.8180 mL | |
| 10 mM | 0.3409 mL | 1.7045 mL | 3.4090 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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