| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
The primary target of rat Kisspeptin-10 is the KISS1 receptor (KISS1R, formerly GPR54), a G protein-coupled receptor belonging to the rhodopsin family. Upon Kp-10 binding, KISS1R primarily activates the phospholipase C signaling pathway, leading to inositol trisphosphate accumulation and intracellular calcium mobilization, while also activating ERK1/2 and p38 mitogen-activated protein kinase pathways. This receptor system plays a critical role in the neuroendocrine control of the hypothalamic-pituitary-gonadal axis. As a potent vasoconstrictor and angiogenesis inhibitor, Kp-10 also exerts anti-angiogenic effects by inhibiting Sp1-mediated VEGF expression and FAK/Rho GTPase activation.
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|---|---|
| ln Vitro |
In vitro studies demonstrate that rat Kisspeptin-10 activates intracellular calcium responses in KISS1R-transfected CHO cells with an EC₅₀ value of 1.54–2.6 × 10⁻⁸ M. Structure-activity relationship studies reveal that Kp-10 exhibits a helicoidal structure between Asn4 and Tyr10 residues (mixed α- and 3₁₀-helix characteristics). Alanine substitutions at position 6 (phenylalanine) or position 10 (tyrosine) result in a significant increase in EC₅₀ values (>6.46 × 10⁻⁶ M), a substantial decrease in the proportion of responsive cells, and a marked increase in the time required to reach maximal response. Ala10 substitution completely disrupts the helical structure in the C-terminal region, indicating that positions 6 and 10 are critical for Kp-10 binding and functional activity at KISS1R.
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| ln Vivo |
Rat Kisspeptin-10 potently stimulates the hypothalamic-pituitary-gonadal axis in vivo. In rodents, Kp-10 administration significantly increases luteinizing hormone secretion, an effect mediated through stimulation of gonadotropin-releasing hormone release. Structure-activity studies show that Ala6 substitution diminishes, while Ala10 substitution completely eliminates LH secretory responses, and co-administration of these antagonist analogs does not affect the LH-releasing ability of Kp-10 itself. In a rat model of methotrexate-induced reproductive toxicity, Kisspeptin-10 (50 nmol/kg, intraperitoneal injection for 10 days) significantly ameliorates decreased sperm motility and abnormal sperm morphology, reduces malondialdehyde levels in testicular tissue, and restores reproductive organ weights. These results indicate that Kp-10 acts as a potential antioxidant to alleviate drug-induced reproductive toxicity.
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| Enzyme Assay |
Cell-free receptor binding studies for rat Kisspeptin-10 primarily utilize radioligand binding assays. A typical protocol includes: 1) Prepare membrane suspensions expressing KISS1R (using native tissues or KISS1R-transfected cells); 2) Dissolve rat Kp-10 in binding buffer (containing 50 mM HEPES pH 7.4, 5 mM MgCl₂, 0.1% BSA) to prepare serial concentrations (e.g., 10⁻¹¹ to 10⁻⁶ M); 3) Add radiolabeled Kp-10 analog (e.g., [¹²⁵I]-Kp-10) and incubate for 60 minutes at room temperature; 4) Terminate the reaction by rapid vacuum filtration and wash filters with ice-cold buffer to remove unbound ligand; 5) Measure membrane-bound radioactivity using a gamma counter; 6) Calculate IC₅₀ and Kᵢ values from competition binding curves. Structure-activity relationship studies can employ alanine scanning mutagenesis to evaluate the impact of key amino acid residues on receptor binding.
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| Cell Assay |
The in vitro cell assay protocol for rat Kisspeptin-10 is as follows: 1) Seed target cells (such as CHO cells stably transfected with rat KISS1R) in culture plates and culture to appropriate density at 37°C with 5% CO₂; 2) Serum-starve cells prior to the experiment (typically using serum-free medium for 4-6 hours); 3) Stimulate cells with various concentrations of rat Kp-10 (typically 10⁻¹¹ to 10⁻⁶ M); 4) For calcium mobilization assays, label cells with fluorescent probes (e.g., Fluo-4 AM) and monitor fluorescence intensity changes in real-time after Kp-10 stimulation using a fluorescence microscope or fluorescence plate reader, recording the percentage of responsive cells and the time to reach maximal response; 5) Calculate EC₅₀ values by nonlinear regression analysis. Under typical conditions, wild-type Kp-10 reaches maximal calcium response within 2 minutes, while less active mutants require over 6 minutes.
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| Animal Protocol |
The in vivo animal assay protocol for rat Kisspeptin-10 is as follows: 1) Use adult male rats (such as Wistar or SD rats); 2) Group assignment: randomly divide animals into vehicle control and Kp-10 treatment groups (multiple doses, e.g., 50 nmol/kg); 3) Dosing regimen: intraperitoneal injection once daily for 10 consecutive days; intravenous injection can also be used for acute pharmacodynamic studies; 4) For methotrexate-induced reproductive toxicity models, initiate Kp-10 treatment 3 days after MTX administration (single dose of 20 mg/kg intraperitoneally); 5) Sample collection: euthanize animals after treatment and collect blood, testes, epididymides, and seminal vesicles; 6) Endpoints: assess sperm motility and abnormal sperm morphology; measure malondialdehyde levels in tissue homogenates as an oxidative stress marker; weigh reproductive organs; 7) Assess hypothalamic-pituitary-gonadal axis activation by measuring serum LH and FSH levels via radioimmunoassay.
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| ADME/Pharmacokinetics |
Rat Kisspeptin-10 exhibits extremely rapid pharmacokinetics in vivo. Stability studies demonstrate that the decomposition half-life of this peptide in rat plasma is only 1.7 minutes at 37°C, 2.9 minutes at 25°C, and 6.8 minutes at 4°C; the principal decomposition product is the N-terminal tyrosine-deleted des-Tyr1-Kp-10. Following intravenous bolus administration of 1.0 mg/kg Kp-10, low ng/mL levels are detectable in rat plasma only during the first few minutes, becoming undetectable below the LLOQ (0.5 ng/mL) by 30 minutes post-dose. Due to the rapid clearance of the peptide in vivo, increased LH levels are commonly used as a surrogate marker for Kp-10 exposure. Rat Kp-10 has a solubility of 50 mg/mL (37.92 mM) in DMSO, and it is recommended to store the powder at -20°C for 3 years or solutions at -80°C for 6 months.
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| Toxicity/Toxicokinetics |
According to the Material Safety Data Sheet, rat Kisspeptin-10 is classified as a non-hazardous substance or mixture with no GHS hazard classification identified. This product is for research use only and is not intended for human or veterinary use. In animal studies, no significant toxicity has been observed following intraperitoneal administration of 50 nmol/kg for 10 consecutive days, with animals showing good tolerance. In the methotrexate-induced reproductive toxicity model, Kp-10 treatment not only showed no toxicity but also alleviated MTX-induced reproductive damage as a potential antioxidant. For handling and storage, avoid inhalation and contact with skin, eyes, and clothing; appropriate personal protective equipment (such as rubber gloves and protective clothing) is recommended, and operations should be performed in well-ventilated areas.
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| References |
| Molecular Formula |
C63H83N17O15
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|---|---|
| Molecular Weight |
1318.43803334236
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| Exact Mass |
1317.625
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| CAS # |
478507-53-8
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| Related CAS # |
Kisspeptin-10, human;374675-21-5;Kisspeptin-10, rat TFA
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| PubChem CID |
90488921
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
-2.4
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| Hydrogen Bond Donor Count |
19
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| Hydrogen Bond Acceptor Count |
17
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| Rotatable Bond Count |
38
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| Heavy Atom Count |
95
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| Complexity |
2630
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| Defined Atom Stereocenter Count |
9
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| SMILES |
CC(C)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N)NC(=O)CNC(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC3=CNC4=CC=CC=C43)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC5=CC=C(C=C5)O)N
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| InChi Key |
HVPGTDOCSYBNFC-INXYWQKQSA-N
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| InChi Code |
InChI=1S/C63H83N17O15/c1-33(2)23-45(58(91)74-43(13-8-22-70-63(68)69)57(90)75-44(54(67)87)25-36-16-20-39(83)21-17-36)73-53(86)31-72-56(89)46(26-34-9-4-3-5-10-34)77-62(95)50(32-81)80-61(94)49(29-52(66)85)79-59(92)47(27-37-30-71-42-12-7-6-11-40(37)42)78-60(93)48(28-51(65)84)76-55(88)41(64)24-35-14-18-38(82)19-15-35/h3-7,9-12,14-21,30,33,41,43-50,71,81-83H,8,13,22-29,31-32,64H2,1-2H3,(H2,65,84)(H2,66,85)(H2,67,87)(H,72,89)(H,73,86)(H,74,91)(H,75,90)(H,76,88)(H,77,95)(H,78,93)(H,79,92)(H,80,94)(H4,68,69,70)/t41-,43-,44-,45-,46-,47-,48-,49-,50-/m0/s1
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| Chemical Name |
(2S)-N-[(2S)-1-[[(2S)-4-amino-1-[[(2S)-1-[[(2S)-1-[[2-[[(2S)-1-[[(2S)-1-[[(2S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-2-oxoethyl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]-2-[[(2S)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]butanediamide
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| Synonyms |
Kisspeptin-10, rat; 478507-53-8; Kisspeptin-10 (mouse, rat); Kisspeptin 10 (rat); CHEMBL3422406;
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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 | 0.7585 mL | 3.7924 mL | 7.5847 mL | |
| 5 mM | 0.1517 mL | 0.7585 mL | 1.5169 mL | |
| 10 mM | 0.0758 mL | 0.3792 mL | 0.7585 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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