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| Targets |
The primary targets of Crustacean Cardioactive Peptide are G protein-coupled receptors (GPCRs) and G protein-coupled receptor kinases (GRKs) found in cardiac and muscle cells. As a neuropeptide, CCAP binds to specific GPCRs on the surface of target cells, triggering intracellular signaling cascades that regulate heart contraction, circulation, and muscle activity. The peptide's cyclic structure, stabilized by a disulfide bridge between cysteine residues at positions 3 and 9, is essential for its receptor binding and biological activity. CCAP's highly conserved sequence across arthropod species suggests that its receptor-mediated mechanisms are evolutionarily conserved.
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| ln Vitro |
While abdominal-A and ultrabithorax are not necessary for the specification of CCAP interneurons, they are necessary to stop CCAP efferent neurons from dying from apoptosis [2].
In vitro, Crustacean Cardioactive Peptide (CCAP) is used as a research tool to study neuromodulation, cardiac function, and muscle contraction in arthropod models. The peptide is known to regulate heartbeat and muscle activity in a concentration-dependent manner in isolated tissue preparations. Its free acid form provides a native-like structure suitable for biochemical assays, receptor studies, and comparative physiology. CCAP also serves as a model peptide for exploring evolutionary conservation in neuropeptide families and dissecting mechanisms underlying rhythmic motor control across invertebrate species. The peptide's activity can be assessed using electrophysiological recordings or muscle contraction assays in vitro. |
| ln Vivo |
The mosquito auxiliary pulsatile organ (APO), heart contraction rate, and hemolymph flow rate in the antennal region are all increased by CCAP, FMRFamide, and SALDKNFMRFamide [1].
In vivo, Crustacean Cardioactive Peptide (CCAP) acts as a cardiac gas pedal, regulating heartbeat and circulation in crustaceans and other arthropods. It modulates neuronal activity and is involved in the coordination of feeding behavior and ecdysis (molting) in arthropods such as Leptinotarsa decemlineata. The peptide's in vivo effects are mediated through its action on GPCRs in cardiac and muscle cells. CCAP is commonly used in neurophysiology and cardiac function studies in invertebrate models to investigate the role of neuropeptides in regulating rhythmic motor patterns and behavior. |
| Enzyme Assay |
Cell-free receptor binding assays for Crustacean Cardioactive Peptide (CCAP) can be performed using membrane preparations from tissues expressing CCAP receptors, such as cardiac or muscle tissue from crustaceans. A typical protocol involves incubating radiolabeled CCAP with membrane preparations in the presence or absence of unlabeled CCAP at various concentrations in a binding buffer. Bound and free ligand are separated by filtration or centrifugation, and the radioactivity is measured by scintillation counting. The binding affinity (Kd) and receptor density (Bmax) are determined by Scatchard analysis. Alternatively, surface plasmon resonance (SPR) can be used to study the interaction of CCAP with its recombinant GPCR.
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| Cell Assay |
For in vitro cellular experiments, cells expressing CCAP receptors (e.g., primary cultures of crustacean cardiac or muscle cells) are cultured in appropriate media and treated with CCAP at various concentrations (typically 1 nM to 10 uM). The peptide's ability to activate its receptor is assessed by measuring downstream signaling, such as intracellular calcium mobilization using fluorescent calcium indicators (e.g., Fluo-4) or cAMP accumulation using ELISA or radioimmunoassay. The duration of treatment varies depending on the specific assay. For electrophysiological studies, the peptide is applied to isolated tissue preparations, and changes in muscle contraction or neuronal firing are recorded.
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| Animal Protocol |
In vivo animal experiments with Crustacean Cardioactive Peptide (CCAP) are typically performed in crustacean or insect models. A common protocol involves injecting the peptide into the hemolymph of the animal or applying it directly to the pericardial organs. The peptide's effects on heart rate, circulation, and muscle activity are monitored using non-invasive techniques such as impedance cardiography or video microscopy. For behavioral studies, the peptide is injected into the animal, and changes in feeding behavior, locomotion, or ecdysis are observed. The dosage and route of administration vary depending on the species and the specific experimental objectives.
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| ADME/Pharmacokinetics |
Crustacean Cardioactive Peptide (CCAP) has a molecular weight of 959.10 g/mol and a molecular formula of C42H58N10O12S2. As a peptide, its pharmacokinetic properties are characterized by rapid clearance from the circulation due to proteolytic degradation. The peptide is typically administered by injection in preclinical studies and has a short half-life in biological fluids. It is stable in lyophilized form and should be stored at -20degC. The peptide is soluble in aqueous buffers and is typically prepared fresh before use. Its pharmacokinetic properties make it suitable for acute pharmacological studies but less suitable for chronic dosing.
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| Toxicity/Toxicokinetics |
The toxicity profile of Crustacean Cardioactive Peptide (CCAP) has not been extensively characterized, as it is a naturally occurring neuropeptide used primarily as a research tool. As a peptide, it is expected to have low toxicity at pharmacological doses. However, high doses may cause alterations in heart rate, circulation, and muscle activity due to its role in regulating these physiological processes. The compound should be handled with standard laboratory precautions and is intended for research use only.
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| References |
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| Additional Infomation |
Crustacean Cardioactive Peptide (CCAP) is a highly conserved, amidated cyclic nonapeptide with the primary structure PFCNAFTGC-NH2 and a disulfide bridge between Cys3 and Cys9. It was first isolated from the pericardial organs of the shore crab Carcinus maenas and acts as a regulator of heartbeat in crustaceans. CCAP also modulates neuronal activity in other arthropods and acts as a stimulator of feeding and a regulator of ecdysis. The peptide targets GPCRs in cardiac and muscle cells and is widely used as a research tool in neurophysiology, cardiac function studies, and comparative physiology. CCAP is available as a research compound and is not approved for clinical use.
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| Molecular Formula |
C42H58N10O12S2
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| Molecular Weight |
959.10
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| Exact Mass |
958.367
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| CAS # |
309247-84-5
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| PubChem CID |
166638345
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| Appearance |
White to off-white solid powder
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| LogP |
-3.9
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| Hydrogen Bond Donor Count |
14
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| Hydrogen Bond Acceptor Count |
15
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| Rotatable Bond Count |
26
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| Heavy Atom Count |
66
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| Complexity |
1720
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| Defined Atom Stereocenter Count |
9
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| SMILES |
C1C(C[C@H](NC([C@@H](NC(=O)[C@H](CC(N)=O)NC([C@H](CS)NC(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@@H]2CCCN2)=O)C)=O)C(N[C@H](C(NCC(N[C@@H](CS)C(O)=O)=O)=O)[C@@H](C)O)=O)=CC=CC=1
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| InChi Key |
ONSAAPXCOZQERN-HNGYRCOKSA-N
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| InChi Code |
InChI=1S/C42H58N10O12S2/c1-22(35(56)48-28(17-25-12-7-4-8-13-25)39(60)52-34(23(2)53)41(62)45-19-33(55)47-31(21-66)42(63)64)46-37(58)29(18-32(43)54)50-40(61)30(20-65)51-38(59)27(16-24-10-5-3-6-11-24)49-36(57)26-14-9-15-44-26/h3-8,10-13,22-23,26-31,34,44,53,65-66H,9,14-21H2,1-2H3,(H2,43,54)(H,45,62)(H,46,58)(H,47,55)(H,48,56)(H,49,57)(H,50,61)(H,51,59)(H,52,60)(H,63,64)/t22-,23+,26-,27-,28-,29-,30-,31-,34-/m0/s1
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| Chemical Name |
(2R)-2-[[2-[[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-4-oxo-2-[[(2R)-2-[[(2S)-3-phenyl-2-[[(2S)-pyrrolidine-2-carbonyl]amino]propanoyl]amino]-3-sulfanylpropanoyl]amino]butanoyl]amino]propanoyl]amino]-3-phenylpropanoyl]amino]-3-hydroxybutanoyl]amino]acetyl]amino]-3-sulfanylpropanoic acid
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
H2O : ~0.67 mg/mL (~0.70 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.61 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 (2.61 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 (2.61 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 | 1.0426 mL | 5.2132 mL | 10.4264 mL | |
| 5 mM | 0.2085 mL | 1.0426 mL | 2.0853 mL | |
| 10 mM | 0.1043 mL | 0.5213 mL | 1.0426 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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