| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
GPR54[1]; Angiogenesis[1]
The primary target of Kisspeptin-10 is the KISS1 receptor (KISS1R), previously known as GPR54, AXOR12, or hOT7T175. KISS1R is a G protein-coupled receptor belonging to the rhodopsin family, widely distributed in tissues including the hypothalamus, pituitary, placenta, pancreas, testis, liver, and small intestine. Upon KP-10 binding, KISS1R primarily activates the phospholipase C pathway, leading to inositol trisphosphate (IP₃) accumulation and intracellular calcium mobilization, while also activating ERK1/2 and p38 mitogen-activated protein kinase signaling pathways. The receptor is also involved in G protein-coupled receptor signaling, cytoskeleton organization regulation, positive regulation of luteinizing hormone secretion, and MAPK cascade activation. Kisspeptin-10, human TFA targets the kisspeptin receptor (KISS1R/GPR54), a GPCR that plays a central role in the regulation of the hypothalamic-pituitary-gonadal axis. Binding of the peptide to GPR54 activates Gq/11 signaling, resulting in phospholipase C activation, IP3-mediated calcium release, and stimulation of GnRH secretion. The kisspeptin-GPR54 signaling pathway also suppresses tumor metastasis through its effects on cell adhesion and migration. |
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| ln Vitro |
The receptor GPR54 and kisspeptin-10 (KP-10) play a crucial role in controlling the release of GnRH in humans and other mammals. GPR54 binds to the protein kisspeptin-10. Kisspeptin-10 is a metastasis suppressor in breast cancer cells and reduces pulmonary human melanoma when activated[1].
In vitro studies demonstrate that Kisspeptin-10 significantly inhibits human umbilical vein endothelial cell migration, invasion, and tube formation, processes critical for angiogenesis. In HTR8/SVneo trophoblast cells treated with high glucose (mimicking gestational diabetes conditions), Gpr54 expression is downregulated with impaired glucose uptake and insulin resistance; Kp-10 treatment restores the cAMP/PKA signaling pathway and enhances glucose uptake by upregulating Glut-4, Insr, and Irs1 expression. Kp-10 also inhibits trophoblast invasion and migration (critical regulatory processes during embryo implantation) and suppresses the gelatinolytic activity of isolated trophoblasts. In KISS1R-transfected CHO cells, human Kp-10 exhibits an EC₅₀ value of approximately 1.54-2.6 × 10⁻⁸ M for Ca²⁺ response activation. In cell-free assays, Kisspeptin-10 binds to purified human GPR54 in membrane preparations with high affinity (Kd typically in the sub-nanomolar range). The binding is measured by radioligand displacement using [125I]-kisspeptin-10, with a Ki of 0.5-1.5 nM for human GPR54. The peptide is highly selective; it does not bind significantly to other GPCRs at nanomolar concentrations, as confirmed by broad GPCR panel screening. |
| ln Vivo |
LH secretion increases in a dose-dependent manner when kisspeptin-10 (7.5, 35, and 100 nM) is infused intravenously. The GnRH antagonist cetrorelix prevents kisspeptin-10 (100 n nM) from stimulating LH production, preventing a single action on gonadotropes[4]. In vivo, kisspeptin-10 suppresses angiogenesis. By preventing tumor angiogenesis, Kp-10 suppresses tumor growth in SCID mice xenografted with human prostate cancer cells (PC-3)[5].
Kisspeptin-10 exhibits significant in vivo activity in various animal models. Intravenous injection of 1 μg/kg Kp-10 increases serum LH levels in healthy men from 4.1 ± 0.4 IU/L to 12.4 ± 1.7 IU/L within 30 minutes; continuous infusion at 1.5 μg/kg/h for 22.5 hours increases LH pulse frequency from 0.7 ± 0.1 to 1.0 ± 0.2 pulses/hour and pulse secretory mass from 3.9 ± 0.4 to 12.8 ± 2.6 IU/L. In a rat model of gestational diabetes mellitus, Kp-10 treatment (intravenous, once daily) improves fasting blood glucose levels, insulin sensitivity, and fetal outcomes, including increased fetal weight and decreased fetal blood glucose. In SCID mice xenografted with human prostate cancer cells (PC-3), Kp-10 inhibits tumor growth by suppressing tumor angiogenesis. In both chicken chorioallantoic membrane assays and VEGF-induced mouse corneal micropocket assays, Kp-10 significantly inhibits angiogenesis in vivo. In cell-based assays using HEK293 cells expressing human GPR54, Kisspeptin-10, human TFA (0.1-100 nM) induces robust calcium mobilization (EC50 typically 0.5-2 nM), increases intracellular IP3 accumulation, and activates ERK1/2 MAP kinase signaling. In breast cancer cell lines, the peptide inhibits cell migration and invasion in transwell assays at 10-100 nM concentrations, and it suppresses pulmonary metastasis in mouse models. The peptide also stimulates GnRH secretion from hypothalamic neurons. |
| Enzyme Assay |
Cell-free receptor binding studies for 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 Kp-10 in binding buffer (containing 50 mM HEPES pH 7.4, 5 mM MgCl₂, 0.1% BSA) to prepare serial concentrations; 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 show that Kp-10 exhibits a helicoidal structure between residues Asn4 and Tyr10 (mixed α- and 3₁₀-helix characteristics), with alanine substitutions at positions 6 or 10 significantly reducing receptor affinity and functional activity.
The binding affinity of Kisspeptin-10 for GPR54 is measured using a cell-free radioligand binding assay. Membranes from CHO or HEK293 cells expressing human GPR54 are incubated with 0.1 nM [125I]-kisspeptin-10 and increasing concentrations (0.0001-1000 nM) of unlabeled Kisspeptin-10, human TFA. After 60-90 minutes at room temperature, bound and free tracers are separated by rapid filtration through GF/C filters pre-soaked in 0.3% polyethyleneimine. Radioactivity is counted, and Ki values are calculated using nonlinear regression analysis. |
| Cell Assay |
The in vitro cell assay protocol for Kisspeptin-10 is as follows: 1) Seed target cells (such as KISS1R-transfected CHO cells, HTR8/SVneo trophoblast cells, HUVEC endothelial cells, or primary trophoblasts) in culture plates and culture to appropriate density at 37°C with 5% CO₂; 2) Treat cells with various concentrations of Kp-10 (typically 10⁻¹¹ to 10⁻⁶ M); 3) For Ca²⁺ 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 plate reader; 4) For migration and invasion assays, use Transwell chambers (8 μm pore size), seed cells in the upper chamber with Kp-10-containing medium in the lower chamber, incubate, then stain and count cells that migrated through the membrane; 5) For tube formation assays, seed HUVECs on Matrigel-coated plates, add Kp-10, incubate for 6-18 hours, and observe and count tube-like structures under a microscope; 6) Detect expression and phosphorylation levels of relevant signaling pathway proteins by Western blot.
HEK293 cells stably expressing human GPR54 are seeded in 96-well plates and loaded with the calcium-sensitive fluorescent dye Fluo-4 AM (2 uM) in HBSS buffer containing 20 mM HEPES (pH 7.4) for 30-60 minutes at 37degC. Varying concentrations (0.0001-1000 nM) of Kisspeptin-10, human TFA are injected by an automated injector, and fluorescence intensity (Ex 485 nm/Em 525 nm) is measured in real-time. The EC50 is calculated from the maximum change in fluorescence relative to baseline using a 4-parameter logistic equation. |
| Animal Protocol |
The in vivo animal assay protocol for Kisspeptin-10 is as follows: 1) Select appropriate animal models: SD rats (gestational diabetes model), SCID mice (tumor xenograft model), Beagle dogs (toxicology studies), or male monkeys (reproductive studies); 2) Group assignment: randomly divide animals into vehicle control and multiple Kp-10 dose groups (e.g., 30, 100, and 1000 μg/kg in dog toxicology studies); 3) Administration routes: intravenous injection, subcutaneous injection, or intracerebroventricular injection; dosing volume typically 2.6 mL/kg, once daily for 14 consecutive days; 4) Sample collection: collect blood samples at multiple time points post-administration (e.g., 5, 15, 30 minutes) and measure LH levels as a pharmacodynamic marker; 5) Behavioral testing: assess anxiety-like behavior and locomotor activity using open field test and novel tank test; 6) Tissue collection: euthanize animals at study termination and collect major organs (such as liver, kidney, spleen, brain, placenta) for histopathological examination; 7) Data acquisition: measure body weight, food consumption, body temperature, electrocardiogram, and respiratory rate.
In mouse models of breast cancer metastasis (e.g., MDA-MB-231 breast cancer cells expressing luciferase injected via tail vein), mice are treated with Kisspeptin-10, human TFA (e.g., 10-100 ug/kg) via intraperitoneal or intravenous injection daily for 3-4 weeks. Metastasis burden is monitored by bioluminescence imaging. The peptide significantly reduces the number and size of pulmonary metastases. Tumor cell migration is also assessed in ex vivo lung tissue sections. Controls include a scrambled kisspeptin peptide. |
| ADME/Pharmacokinetics |
Kisspeptin-10 exhibits extremely rapid pharmacokinetics in vivo. In in vitro stability studies, the decomposition half-life of Kp-10 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 primary 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 of Kp-10 are detectable in rat plasma only during the first few minutes, becoming undetectable by 30 minutes post-dose. In humans, the terminal half-life of Kp-10 after intravenous infusion is approximately 3.8-4.1 minutes. Following subcutaneous administration of radiolabeled Kp-10 analogs in dogs, approximately complete recovery of the administered radioactive dose is achieved within 48-72 hours, primarily excreted in urine after extensive metabolism. Due to the rapid clearance of the peptide in vivo, LH levels are commonly used as a surrogate marker for Kp-10 exposure in studies.
Kisspeptin-10, human TFA has a molecular weight of 1416.46 Da (free base) and molecular formula C63H83N17O14 plus TFA. The TFA salt enhances solubility and storage stability. The peptide has a short plasma half-life (minutes) due to rapid proteolytic degradation; for sustained effects, continuous infusion or frequent daily injections are required. The peptide is soluble in sterile water at concentrations up to 1 mg/mL and should be stored as a lyophilized powder at -20degC, protected from light. |
| Toxicity/Toxicokinetics |
Kisspeptin-10 demonstrates a favorable safety profile in preclinical studies. In a 14-day repeat-dose GLP toxicology study in Beagle dogs, once-daily intravenous administration of KP-10 at dose levels of 30, 100, and 1000 μg/kg for 14 days revealed no overt signs of drug-related toxicity. No significant toxicity was observed in clinical signs, body weights, food consumption, clinical pathology, histopathology, urinalysis, electrocardiogram, or respiratory rate. The 1000 μg/kg dose was established as the No Observed Adverse Effect Level (NOAEL). The study also noted a 20-day rat TK study suggesting potential toxicity to skeletal development in juvenile animals, thus the safety of Kp-10 in juvenile animals requires further evaluation. In younger (4-week-old) rats, MCH (monomethylhistamine) levels were slightly increased in female animals. In human studies, no serious adverse events have been reported following Kp-10 administration, with common adverse reactions primarily being mild injection site reactions.
Kisspeptin-10 is well-tolerated at typical research doses in animals. In the SDS, Kisspeptin-10, human TFA is classified as not a hazardous substance or mixture. However, standard laboratory safety precautions should be followed: avoid inhalation, contact with eyes and skin, and use only in areas with appropriate exhaust ventilation. At high doses, vasoconstrictor effects may cause changes in blood pressure and heart rate. No significant organ toxicity has been reported. |
| References |
[1]. Kisspeptin-10 (KP-10) stimulates osteoblast differentiation through GPR54-mediated regulation of BMP2 expression and activation. Sci Rep. 2018 Feb 1;8(1):2134.
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| Additional Infomation |
Kisspeptin-10, human TFA is a research peptide not approved for clinical use. It is derived from the product of the KiSS1 gene, which was originally identified as a human melanoma metastasis suppressor. The peptide activates the GPR54 receptor, which is also known as the kisspeptin receptor. Kisspeptin-10 has been studied for its potential applications in reproductive medicine (e.g., treating infertility, hypogonadotropic hypogonadism), as well as in oncology (limiting metastasis). It is also a potent vasoconstrictor.
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| Molecular Formula |
C63H83N17O14.C2HF3O2
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|---|---|
| Molecular Weight |
1416.46
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| Exact Mass |
1415.6234041
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| Related CAS # |
Kisspeptin-10, human;374675-21-5
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| PubChem CID |
155977641
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| Sequence |
H-Tyr-Asn-Trp-Asn-Ser-Phe-Gly-Leu-Arg-Phe-NH2.TFA
L-tyrosyl-L-asparagyl-L-tryptophyl-L-asparagyl-L-seryl-L-phenylalanyl-glycyl-L-leucyl-L-arginyl-L-phenylalaninamide trifluoroacetic acid |
| SequenceShortening |
YNWNSFGLRF-NH2
YNWNSFGLRF |
| Appearance |
White to off-white solid at room temperature
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| Hydrogen Bond Donor Count |
19
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| Hydrogen Bond Acceptor Count |
21
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| Rotatable Bond Count |
38
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| Heavy Atom Count |
101
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| Complexity |
2670
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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=CC=C1)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.C(=O)(C(F)(F)F)O
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| InChi Key |
TYHMUNFOLJTKBC-QDUZOICUSA-N
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| InChi Code |
InChI=1S/C63H83N17O14.C2HF3O2/c1-34(2)24-45(58(90)74-43(18-11-23-70-63(68)69)57(89)75-44(54(67)86)26-35-12-5-3-6-13-35)73-53(85)32-72-56(88)46(27-36-14-7-4-8-15-36)77-62(94)50(33-81)80-61(93)49(30-52(66)84)79-59(91)47(28-38-31-71-42-17-10-9-16-40(38)42)78-60(92)48(29-51(65)83)76-55(87)41(64)25-37-19-21-39(82)22-20-37;3-2(4,5)1(6)7/h3-10,12-17,19-22,31,34,41,43-50,71,81-82H,11,18,23-30,32-33,64H2,1-2H3,(H2,65,83)(H2,66,84)(H2,67,86)(H,72,88)(H,73,85)(H,74,90)(H,75,89)(H,76,87)(H,77,94)(H,78,92)(H,79,91)(H,80,93)(H4,68,69,70);(H,6,7)/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-1-oxo-3-phenylpropan-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;2,2,2-trifluoroacetic acid
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| Synonyms |
Kisspeptin-10 human TFA; Kisspeptin-10, human (TFA); orb1296647; KISSPEPTIN-10, HUMAN TFA; ...; Kisspeptin-10, human (TFA)(374675-21-5,FREE);
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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 :~100 mg/mL (~70.60 mM)
H2O :~4 mg/mL (~2.82 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 | 0.7060 mL | 3.5299 mL | 7.0599 mL | |
| 5 mM | 0.1412 mL | 0.7060 mL | 1.4120 mL | |
| 10 mM | 0.0706 mL | 0.3530 mL | 0.7060 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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