| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
αvβ3
Cyclo(Arg-Gly-Asp-D-Phe-Cys) TFA specifically targets the alphavbeta3 integrin, a cell surface heterodimeric receptor that is overexpressed on activated endothelial cells during angiogenesis and on various cancer cells (e.g., glioblastoma, melanoma, breast cancer). It binds with high affinity to the alphavbeta3 receptor, thereby disrupting the interaction between the integrin and its natural extracellular matrix ligands (e.g., vitronectin, fibronectin). This competitive antagonism blocks integrin-mediated cell adhesion, migration, and pro-survival signaling pathways. |
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| ln Vitro |
The mESC transcription factors Oct 4, Sox 2, and Nanog are down-regulated by Cyclo(Arg-Gly-Asp-D-Phe-Cys) TFA (0.5 mM; 24 h) [1]. Construction of mESC-col I (type I collagen) is inhibited by Cyclo(Arg-Gly-Asp-D-Phe-Cys) TFA (0.5 mM) integrin gene expression [1]. After being exposed to Cyclo(Arg-Gly-Asp-D-Phe-Cys) TFA (0.5 mM), mESCs aggregated and separated from the surface [1].
Cyclo(RGDfC) TFA potently inhibits the adhesion of alphavbeta3-expressing cells to vitronectin-coated surfaces. At nanomolar concentrations (IC50 typically <10 nM), the peptide disrupts integrin-mediated cell spreading and focal adhesion formation. It also inhibits the pluripotency gene expression (e.g., Nanog, Oct4, Sox2) of mouse embryonic stem cells (mESCs) in culture, indicating a role for integrin signaling in maintaining the stem cell state. |
| ln Vivo |
Mice lacking leukemia inhibitory factor (LIF; injection in both legs) that are treated with cyclo(Arg-Gly-Asp-D-Phe-Cys) TFA (0.5 mM, 24 h)-treated mESCs suppress severe combined immunity and develop teratomas, indicating that integrin interactions are necessary for the process of mESC tumor formation in vivo [1].
Cyclo(Arg-Gly-Asp-D-Phe-Cys) TFA suppresses the tumorigenic potential of mESCs in immunocompromised mice, inhibiting the formation of teratomas. In various orthotopic xenograft and syngeneic mouse tumor models (e.g., breast cancer, melanoma, glioblastoma), systemic or intratumoral administration of RGD-based cyclic peptides (often conjugated to imaging agents or drugs) leads to reduced tumor growth, metastasis, and angiogenesis. When conjugated to cytotoxic payloads or nanoparticles, the cyclic RGD peptide facilitates targeted drug delivery to alphavbeta3-positive tumor vasculature and cancer cells. |
| Enzyme Assay |
The in vitro binding affinity and specificity of Cyclo(RGDfC) is evaluated using solid-phase binding assays or surface plasmon resonance (SPR). Purified alphavbeta3 integrin protein is immobilized onto a sensor chip or ELISA plate. Increasing concentrations of Cyclo(RGDfC) TFA (0.1-1000 nM) are flowed over the surface in the presence of a constant concentration of labeled vitronectin. The dissociation constant (Kd) or IC50 is calculated. Alternatively, a cell-based competitive adhesion assay is used: alphavbeta3-expressing cells (e.g., U87MG glioblastoma cells) are pre-incubated with peptide (0-1000 nM) for 30 minutes, then seeded onto vitronectin-coated wells. After 30-60 minutes at 37degC, unbound cells are washed away, and bound cells are quantified using crystal violet staining or Calcein-AM fluorescence.
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| Cell Assay |
Cellular assays to assess integrin inhibition: alphavbeta3-expressing cells (e.g., U87MG, A375 melanoma, HUVEC) are cultured in DMEM with 10% FBS. Cells are serum-starved for 4 hours before the assay. For cell adhesion experiments, 96-well plates are coated with vitronectin (5 microg/mL) overnight at 4degC, then blocked with 1% BSA. Cells are harvested, resuspended in serum-free medium containing serial dilutions of Cyclo(RGDfC) TFA (0-1000 nM) for 30 minutes. 1 × 10⁵ cells are added to each well and allowed to adhere for 45 minutes at 37degC. Non-adherent cells are removed by gentle washing with PBS. Adherent cells are fixed with 4% paraformaldehyde, stained with 0.5% crystal violet, and solubilized with 10% acetic acid. Absorbance is read at 590 nm. For cell migration assays, a scratch (wound-healing) assay is performed: a confluent monolayer is scratched with a pipette tip, and migration into the scratch area is monitored for 24-48 hours in the presence or absence of peptide. For invasion, a modified Boyden chamber (Matrigel-coated transwell) is used.
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| Animal Protocol |
The in vivo activity of Cyclo(RGDfC) TFA is frequently studied in the mouse embryonic stem cell (mESC) teratoma model. Female SCID mice are injected subcutaneously with 1 × 10⁶ mESCs resuspended in 100 microL Matrigel. Cyclo(RGDfC) TFA (dissolved in PBS) is administered daily at a dose of 10-50 mg/kg (intraperitoneal or intravenous) starting from the day of tumor cell injection. Control groups receive vehicle (PBS) or a scrambled RGD peptide. Tumor volume (length × width2/2) is measured every 2-3 days for 4-8 weeks. Upon sacrifice, tumors are excised, weighed, and processed for immunohistochemistry using antibodies against the pluripotency markers (Nanog, Oct4) to assess the mechanism of action. The teratoma formation rate and size are significantly reduced in peptide-treated groups. For biodistribution studies, Cyclo(RGDfC) is conjugated to a near-infrared dye (e.g., Cy5.5) and injected i.v. into mice bearing alphavbeta3-positive tumors (e.g., U87MG xenografts). Fluorescence imaging is performed at various time points (1-48 hours) to confirm tumor-specific homing.
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| ADME/Pharmacokinetics |
Cyclic RGD peptides, such as Cyclo(RGDfC), typically have a short plasma half-life in mice (approx. 5-30 minutes) due to rapid renal clearance and some proteolytic degradation despite cyclization. The TFA salt form enhances water solubility for i.v. or i.p. administration. The peptide does not significantly bind to plasma proteins, and its volume of distribution is primarily limited to the extracellular space. The presence of the disulfide bridge (Cys-Cys) is essential for structural stability, and it is stable in plasma for at least 2-4 hours ex vivo. The pharmacokinetics are largely determined by glomerular filtration as the molecular weight is <1500 Da. Conjugation to larger carriers (e.g., polymers, nanoparticles, liposomes) dramatically prolongs circulation time and improves tumor accumulation.
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| Toxicity/Toxicokinetics |
The acute toxicity of Cyclo(RGDfC) TFA in animal models is low. Doses up to 100 mg/kg administered intraperitoneally or intravenously in mice do not result in significant mortality, severe weight loss (>20%), or signs of systemic toxicity (lethargy, ruffled fur, abnormal breathing). At high doses, mild gastrointestinal distress may occur, but there are no major organ histopathological changes. The compound is not known to be genotoxic. Long-term toxicity studies have not been formally reported, but cyclic RGD peptides are generally considered well-tolerated research tools. However, since they inhibit alphavbeta3 integrin (which plays a role in physiological angiogenesis, e.g., during wound healing), prolonged high-dose exposure could potentially delay tissue repair. Standard laboratory precautions apply; avoid inhalation and skin contact.
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| References | |
| Additional Infomation |
Cyclo(Arg-Gly-Asp-D-Phe-Cys) (Cyclo-RGDfC) is a cyclic RGD peptide containing a disulfide bridge between the D-Phe and Cys residues. The presence of D-Phe is crucial for enhancing alphavbeta3 selectivity over alphavbeta5 and other integrins. The cyclization (vs. linear RGD peptides) confers higher binding affinity (10-100 fold increase) and resistance to proteolytic degradation, making it a superior ligand for integrin targeting. This compound is a key research tool for studying integrin-mediated cell adhesion, angiogenesis, tumor progression, and stem cell pluripotency. It is widely used for the functionalization of nanoparticles for targeted drug delivery and for molecular imaging probes (PET, SPECT, NIR fluorescence, MRI) to detect alphavbeta3 expression in tumors. Cyclo(RGDfC) is for research use only, not for human therapeutic or diagnostic use without conjugation to FDA-approved carriers.
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| Molecular Formula |
C26H35F3N8O9S
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|---|---|
| Molecular Weight |
692.66
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| Related CAS # |
Cyclo(Arg-Gly-Asp-D-Phe-Cys);862772-11-0
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| Appearance |
White to off-white solid powder
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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 :~5 mg/mL (~7.22 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 | 1.4437 mL | 7.2185 mL | 14.4371 mL | |
| 5 mM | 0.2887 mL | 1.4437 mL | 2.8874 mL | |
| 10 mM | 0.1444 mL | 0.7219 mL | 1.4437 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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