| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
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| Targets |
AP C5 selectively inhibits guanosine 3',5'-cyclic monophosphate (cGMP)-dependent protein kinase II (cGKII). cGKII is a serine/threonine protein kinase that is activated by cGMP and plays a key role in regulating intestinal ion transport, specifically by activating the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel. By inhibiting cGKII, AP C5 blocks the cGMP-mediated signaling pathway that leads to CFTR activation and subsequent anion secretion in the intestine. This mechanism makes AP C5 a potential therapeutic agent for conditions characterized by excessive intestinal fluid secretion, such as secretory diarrhea.
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| ln Vitro |
Intestinal tissue cGMP-dependent cGKII-mediated protein phosphorylation and cGMP-dependent CFTR-mediated anion secretion can both be efficiently inhibited by AP-C5 [1]. AP-C5 suppresses PDE to improve cAMP signaling [1]. In the mouse ileum, AP-C5 (20 μM) partially inhibits the short-circuit current (Isc) response mediated by heat-stable toxin (STa) [1].
In vitro, AP C5 efficiently inhibits cGMP-dependent cGKII-mediated protein phosphorylation and CFTR-mediated anion secretion in intestinal tissue. It also suppresses PDE to improve cAMP signaling. In isolated mouse ileum preparations, AP C5 at a concentration of 20 μM partially inhibits the short-circuit current (Isc) response mediated by heat-stable toxin (STa). STa is a bacterial toxin that activates guanylate cyclase, leading to increased cGMP levels, activation of cGKII, and CFTR-mediated chloride secretion. The partial inhibition of the Isc response by AP C5 confirms its ability to block this pathway at the level of cGKII. |
| ln Vivo |
In vivo, AP C5 has been studied for its potential to reduce diarrheal symptoms. While specific in vivo data for AP C5 are limited, its mechanism of action suggests that it could be effective in animal models of secretory diarrhea. By inhibiting cGKII in the intestine, AP C5 would be expected to reduce CFTR-mediated fluid secretion, thereby alleviating diarrhea. Studies with other cGKII inhibitors have shown efficacy in models of toxin-induced and infectious diarrhea. AP C5's selectivity for cGKII over other kinases makes it a promising candidate for further in vivo evaluation.
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| Enzyme Assay |
Non-cellular in vitro assays for AP C5 involve kinase inhibition studies. A typical protocol uses purified recombinant human cGKII enzyme. The kinase reaction is performed in a buffer containing ATP, a peptide substrate, and varying concentrations of AP C5. The reaction is initiated by the addition of the enzyme and incubated at 30°C for a specified time. The amount of phosphorylated substrate is then quantified using a luminescent or fluorescence-based detection method, such as the ADP-Glo™ Kinase Assay. The IC50 value is determined by plotting the percentage of kinase activity remaining against the log of the inhibitor concentration. The pIC50 is calculated as the negative logarithm of the IC50.
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| Cell Assay |
Cellular assays for AP C5 are performed using intestinal epithelial cell lines or primary intestinal organoids. Cells are treated with AP C5 at various concentrations (e.g., 1-50 μM) for a period of time. To measure cGKII activity, cells are stimulated with a cGMP analog (e.g., 8-Br-cGMP) or a guanylate cyclase agonist (e.g., STa). The level of cGKII-mediated protein phosphorylation is then assessed by Western blotting using phospho-specific antibodies. Alternatively, CFTR-mediated anion secretion can be measured in Ussing chamber experiments using intestinal tissue segments. The short-circuit current (Isc) is monitored, and the inhibitory effect of AP C5 on the Isc response is determined.
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| Animal Protocol |
In vivo animal studies for AP C5 are conducted in mouse models of diarrhea. A common model is the STa-induced diarrhea model, where mice are orally administered STa to induce intestinal fluid secretion. AP C5 is administered either orally or intraperitoneally at doses such as 5-20 mg/kg prior to or concurrent with STa challenge. After a specified period (e.g., 4-6 hours), the animals are euthanized, and the intestinal fluid accumulation is measured. The weight of the entire small intestine is recorded and compared to the remaining body weight to calculate the fluid accumulation ratio. A reduction in this ratio in AP C5-treated animals compared to vehicle controls indicates efficacy.
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| ADME/Pharmacokinetics |
AP C5 is a small molecule with drug-like properties suitable for in vivo studies. It is soluble in DMSO at concentrations up to 25 mg/mL. For in vivo administration, it can be formulated in vehicles such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline. Detailed pharmacokinetic parameters for AP C5, such as half-life, bioavailability, and clearance, have not been fully reported in the available literature. However, given its potency and selectivity, it is likely to have acceptable pharmacokinetic properties for use in animal models.
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| Toxicity/Toxicokinetics |
Detailed toxicological data for AP C5 have not been extensively reported. As a research chemical, it is not intended for human use and is strictly for preclinical research purposes. Standard safety precautions should be followed when handling AP C5, including the use of personal protective equipment and working in a well-ventilated area. The compound is typically stored as a powder at -20°C for up to three years or at 4°C for up to two years. In solution, it is stable for six months at -80°C or one month at -20°C.
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| References |
[1]. Marcel J C Bijvelds, et al. Selective inhibition of intestinal guanosine 3',5'-cyclic monophosphate signaling by small-molecule protein kinase inhibitors. J Biol Chem. 2018 May 25;293(21):8173-8181.
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| Additional Infomation |
AP C5 is a valuable pharmacological tool for studying the role of cGKII in intestinal physiology and disease. Its selectivity for cGKII makes it a useful probe for dissecting cGMP signaling pathways. The compound's alkyne group also makes it amenable to click chemistry, allowing for the creation of probes for target identification and imaging studies. AP C5 is not a clinically approved drug and has not entered clinical trials. Its primary application is in academic and pharmaceutical research to validate cGKII as a therapeutic target for diarrheal diseases.
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| Molecular Formula |
C16H13N5
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| Molecular Weight |
275.307922124863
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| Exact Mass |
275.117
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| CAS # |
2234272-10-5
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| PubChem CID |
137919866
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| Appearance |
Light yellow to light brown solid powder
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| LogP |
2
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
21
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| Complexity |
368
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N1(C=NC=C1)C1C=CC(C2C=CN=C(NCC#C)N=2)=CC=1
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| InChi Key |
QKHQFSQJQKGFAA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H13N5/c1-2-8-18-16-19-9-7-15(20-16)13-3-5-14(6-4-13)21-11-10-17-12-21/h1,3-7,9-12H,8H2,(H,18,19,20)
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| Chemical Name |
4-(4-imidazol-1-ylphenyl)-N-prop-2-ynylpyrimidin-2-amine
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| Synonyms |
AP-C5 APc5AP C5
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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 : ~25 mg/mL (~90.81 mM)
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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 | 3.6323 mL | 18.1613 mL | 36.3227 mL | |
| 5 mM | 0.7265 mL | 3.6323 mL | 7.2645 mL | |
| 10 mM | 0.3632 mL | 1.8161 mL | 3.6323 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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