 Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C16H26CUN6O6 |
| Molecular Weight | 461.96 |
| Exact Mass | 461.12098 |
| Elemental Analysis | C, 41.60; H, 5.67; Cu, 13.76; N, 18.19; O, 20.78 |
| CAS # | 300801-03-0 |
| Related CAS # | Copper tripeptide;89030-95-5 |
| Appearance | Typically exists as solids at room temperature |
| LogP | 0 |
| SMILES | [Cu+2].O=C([C@H](CC1=CN=CN1)NC(CN)=O)N[C@H](C(=O)[O-])CCCCN.O=C([C@H](CC1=CN=CN1)NC(CN)=O)N[C@H](C(=O)[O-])CCCCN |
| Synonyms | GHK-Cu acetate; GHK Cu acetate; 300801-03-0; copper;acetic acid;(2S)-6-amino-2-[[(2S)-2-(2-aminoacetyl)azanidyl-3-(1H-imidazol-4-yl)propanoyl]amino]hexanoate; GHK-Cu; GHK-Cu acetate, Gly-His-Lys-Cu(II) |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets | Tripeptide |
| ln Vitro |
Copper tripeptide (1 nM; 0-96 hours) acetate affects the population doubling time of irradiated fibroblasts, causing it to approach that of controls [1]. Copper tripeptide (1 nM; 0-120 hours) acetate significantly increased the production of basic fibroblast growth factor in 24-hour irradiated fibroblasts compared with normal controls [1].
Irradiated fibroblasts survived and replicated in serum-free media. The population-doubling times of normal and irradiated fibroblasts exposed to GHK-Cu were faster than those of nontreated controls. Irradiated fibroblasts treated with GHK-Cu doubled at a rate that approximated that of untreated controls, and produced significantly more basic fibroblast growth factor and vascular endothelial growth factor than untreated controls early after GHK-Cu exposure. Conclusions: Irradiated fibroblasts survive and replicate in serum-free media, establishing this model as ideal for evaluating growth factor production in vitro. Copper tripeptide accelerates the growth of normal and irradiated fibroblasts to the point where treated irradiated fibroblasts approximate the population-doubling time of normal controls. An early increase in basic fibroblast growth factor and vascular endothelial growth factor production by GHK-Cu-treated irradiated fibroblasts may improve wound healing. [1] |
| Cell Assay |
Experiments were performed with cells in their first or second passage. At the time of experimentation, the fibroblasts were washed with phosphate-buffered saline solution, and 0.05% trypsin was used to release the confluent cells from the flask wall. Trypsin soybean inhibitor (GIBCO) in a 1:1 ratio inactivated the trypsin. Cell culture viability was determined by means of trypan-blue dye exclusion, and cells counts were performed in duplicate using a hemocytometer and phase-contrast microscopy. Cells were then seeded at a density of 5 × 103 (normal) and 3 × 103 (irradiated) cells/well in each well of a sterile 96-well plate using a commercially available serum-free medium. This medium has been shown to sustain dermal fibroblast growth to at least 7 days with greater than 90% viability. At 0 hours, GHK-Cu solution (1 × 10−9 M) in the serum-free medium was added to the treatment groups, and an equalvolume of plain serum-free medium was added to the untreated control groups. Untreated cells from each cell line were used for controls. Cell counts were performed using a cell proliferation assay system with reagent 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) 24, 48, 72, and 96 hours after initiation for growth curve generation. The WST-1 assay is a colorimetric assay used in the quantification of cell proliferation and cell viability based on the cleavage of the tetrazolium salt WST-1 by mitochondrial dehydrogenases in viable cells. It is a nonradioactive alternative to the tritium-thymidine incorporation assay. Assays were read using an automated plate reader. Optical densities were analyzed with commercially available software. Cell counts were determined by comparison with a standard curve derived from known cell quantities calculated for each cell type and medium. At each 24-hour interval, cell-free supernatant was collected from the testing wells in triplicate. Samples were stored at −80°C in microcentrifuge tubes for later growth factor assays. Expression of bFGF, TGF-β1, and VEGF was evaluated for each group by means of a solid-phase enzyme-linked immunosorbent assay at 24-hour intervals. We calculated cell population-doubling times (PDT) from logarithmic best-fit curves.[1] |
| References |
[1]. Effects of copper tripeptide on the growth and expression of growth factors by normal and irradiated fibroblasts. Arch Facial Plast Surg. 2005 Jan-Feb;7(1):27-31. |
| Additional Infomation |
Prezatide is a tripeptide consisting of glycine, histidine, and lysine which readily forms a complex with copper ions. Prezatide is used in cosmetic products for the skin and hair. It is known to aid wound healing and its potential applications in chronic obstructive pulmonary disease and metastatic colon cancer are currently being investigated. Drug Indication Commonly used in cosmetic products for the skin and hair. Mechanism of Action Prezatide in complex with copper increases the synthesis and deposition of type I collagen and glycosaminoglycan. It also increases the expression of matrix metalloproteinase-2 as well as tissue inhibitor of matrix metalloproteinases-1 and -2, suggesting that it plays a role in the modulation of tissue remodeling. It is thought that prezatide's antioxidant activity is due to its ability to supply copper for superoxide dismutase and its anti inflammatory ability due to the blockage the of iron (Fe2+) release during injury. Prezatide also increases angiogenesis to injury sites. The precise mechanisms of these effects are unknown. It is also unknown whether prezatide's effects are due to the action of the tripeptide itself or its ability to localize and transport copper. Prezatide is known to be bound by heparin and heparin sulfate Pharmacodynamics Prezatide in complex with copper improve skin elasticity, density, and firmness, reduces fine lines and wrinkles, reduces photodamage, increases keratinocyte proliferation. Prezatide also displays anti-oxidant and angiogenic effects and appears to modulate tissue remodeling in injury. Objective: To evaluate the effects of copper tripeptide (GHK-Cu) on the growth and autocrine production of basic fibroblast growth factor, transforming growth factor beta1, and vascular endothelial growth factor by normal and irradiated fibroblasts in a serum-free in vitro environment. Methods: Primary human dermal fibroblast cell lines were established after explantation from intraoperative specimens obtained from patients who had undergone radiation therapy for head and neck cancer. Normal and irradiated fibroblasts were propagated in serum- and growth factor-free media. Treatment groups were exposed to GHK-Cu (1 x 10(-9) mol/L). We measured cell counts and production of basic fibroblast growth factor, transforming growth factor beta1, and vascular endothelial growth factor.[1] Several interesting findings are demonstrated. First, survival and growth of irradiated fibroblasts was demonstrated within the serum-free media. To our knowledge, our laboratory is the first to document this phenomenon using irradiated human fibroblasts. Our laboratory has already demonstrated survival and growth of normal, fetal, and keloid fibroblasts in this serum-free environment. Serum-free cell culture is essential when measuring changes in the growth factor milieu and is now a viable model for future studies involving irradiated fibroblasts. Second, the data established differences in the baseline production of growth factors between normal and irradiated fibroblasts in a head-to-head model. Production of bFGF by normal fibroblasts was significantly increased when compared with that of irradiated fibroblasts at all but 1 time point (72 hours). Production of TGF-β1 by normal fibroblasts was significantly increased when compared with that of irradiated fibroblasts at the 24-hour mark. Finally, production of VEGF by normal fibroblasts was significantly increased when compared with that of irradiated fibroblasts at 24 and 48 hours. It is reasonable to suppose that these differences play an influential role in the differing wound-healing properties of these wounds clinically. Third, the data show that modulation of the environment with GHK-Cu is associated with changes in the growth factor milieu. The GHK-Cu–treated irradiated fibroblasts showed significantly greater production of bFGF than controls at 24 and 72 hours. In fact, GHK-Cu–treated irradiated fibroblasts produced significantly more bFGF than normal controls at the 24-hour interval. Furthermore, GHK-Cu–treated irradiated fibroblasts produced significantly more VEGF than normal controls at the 24-hour interval. This finding is of importance given the known benefit of an early presence of these growth factors in the healing wound. Finally, the data show that modulation of the environment with GHK-Cu is associated with a dramatic increase in fibroblast PDT. This was demonstrated in the normal and irradiated cell lines. One striking finding is that population growth in GHK-Cu–treated irradiated fibroblasts assumed that of normal controls. The clinical implications of this are not yet known. However, given the important role of fibroblasts in wound healing, one might hypothesize that more fibroblasts in an irradiated wound bed would lead to a generalized improvement in wound healing. |
Solubility Data
| 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 |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1647 mL | 10.8234 mL | 21.6469 mL | |
| 5 mM | 0.4329 mL | 2.1647 mL | 4.3294 mL | |
| 10 mM | 0.2165 mL | 1.0823 mL | 2.1647 mL |