| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
LyP-1 TFA targets the p32 protein (also known as gC1qR or HABP1), a receptor that is overexpressed on the surface of tumor-associated cells, including cancer cells and tumor-associated macrophages, and is involved in mitochondrial function and cell adhesion.
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| ln Vitro |
LyP-1 TFA is a tumor-homing peptide that selectively binds to p32 receptors with high affinity. Upon binding, it is internalized into cells. This peptide can be used to deliver therapeutic agents or imaging probes specifically to tumors. LyP-1 has been shown to accumulate in tumors and to be taken up by cancer cells.
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| ln Vivo |
In the LyP-1-treated group, LyP-1 TFA (8 weeks) significantly lowers the rates of both plaque occupancy and creation. Furthermore, compared to control peptide, a greater rate of apoptosis is seen in macrophages liberated from hypoxic plaques following LyP-1 treatment[1]. For optical imaging, a near-infrared fluorophore (Cy5.5) is used to identify the LyP-1 peptide. Tumor-bearing BALB/C mice are injected with Cy5.5-LyP-1 (0.8 nmol) through the middle phalanges of their upper extremities on days 3, 7, 14, and 21 following their inoculation with 4T1 cells. At days 3, 7, 14, and 21 following tumor cell inoculation, the fluorescence intensities were 0.024, 0.038, 0.048, and 0.106×106 photon/cm2/sec, respectively. These values are 1.02, 1.63, 2.04, and 4.52-fold greater than in the contralateral LNs. LyP-1 peptide binding specific to lymphatics is suggested by Cy5.5-LyP-1 staining in lymph nodes co-localizing with LYVE-1[1].
In vivo studies have shown that LyP-1 TFA can home to tumors and be internalized by cancer cells. Treatment with LyP-1 TFA for 8 weeks significantly reduces plaque formation and occupancy, accompanied by increased apoptotic rates in macrophages from hypoxic plaques, indicating potential therapeutic applications in atherosclerosis as well as cancer. |
| Enzyme Assay |
Non-cell binding assays for LyP-1 TFA are performed to determine its affinity for recombinant p32 protein. A Surface Plasmon Resonance (SPR) experiment is a typical method. Briefly, recombinant p32 protein is immobilized on a CM5 sensor chip via amine coupling. LyP-1 TFA is prepared in running buffer (10 mM HEPES, 150 mM NaCl, 0.005% Tween-20, pH 7.4) at concentrations ranging from 1 nM to 10 uM. Association (2-3 minutes) and dissociation (5-10 minutes) phases are monitored. The KD is calculated by fitting the sensorgrams to a 1:1 Langmuir binding model. Alternatively, an ELISA binding assay can be performed. 96-well plates are coated with recombinant p32 protein (1-5 microg/mL in PBS) overnight at 4degC. The plates are blocked with 3% BSA or 5% non-fat milk. Serially diluted biotinylated LyP-1 TFA (0.001-1000 nM) is added and incubated for 1-2 hours. After washing, HRP-conjugated streptavidin is added. The signal is developed with TMB substrate and measured at 450 nm. The EC50 is calculated from the dose-response curve. For competitive binding assays, unlabeled LyP-1 TFA competes with a labeled tracer.
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| Cell Assay |
Cellular assays for LyP-1 TFA are performed using p32-positive cancer cell lines such as MDA-MB-231 (breast cancer), HeLa, or A549 (lung cancer), or p32-positive tumor-associated macrophages. For binding and uptake studies, cells are seeded in 96-well plates or on coverslips in 24-well plates (1-5×10⁴ cells/well) and allowed to attach overnight. LyP-1 TFA is fluorescently labeled (e.g., with FITC, Cy5, or rhodamine) via the terminal amine or a linker, or used as a conjugate with nanoparticles/drugs. The fluorescently labeled peptide is added to the cells at concentrations of 0.1-10 microM for 30-120 minutes at 37degC. Cells are washed with cold PBS to remove unbound peptide, and fluorescence intensity is measured using a fluorescence plate reader (excitation/emission depends on the fluorophore), or visualized by confocal microscopy (DAPI for nuclei, LyP-1-fluorophore for peptide). For competition assays, cells are pre-incubated with a 10-100x excess of unlabeled LyP-1 TFA (0.1-1 mM) for 30 minutes prior to adding the labeled peptide. For cytotoxicity assays, cells are treated with LyP-1 TFA (0.1-100 microM) for 24-72 hours, and viability is assessed by MTT or CellTiter-Glo. For apoptosis assays, cells are stained with annexin V-FITC/PI and analyzed by flow cytometry. For drug delivery studies, LyP-1 is conjugated to chemotherapeutic drugs (e.g., doxorubicin, paclitaxel) or nanoparticles. Cells are treated with the conjugate (0.01-10 microM drug equivalent) for 24-72 hours. For macrophage polarization studies, THP-1-derived macrophages are treated with LyP-1 TFA (1-50 microM) and their polarization state is assessed by measuring M1 and M2 markers (CD86, CD206) by flow cytometry.
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| Animal Protocol |
In vivo animal studies are performed in mouse tumor xenograft models (e.g., MDA-MB-231 breast cancer, A549 lung cancer) or in atherosclerosis models (ApoE-/- mice). For tumor targeting studies, 6-8 week old female nude mice are subcutaneously implanted with 5×10⁶ p32-positive tumor cells. When tumors reach 100-200 mm3, fluorescently labeled LyP-1 TFA (0.1-2 mg/kg) is administered intravenously via the tail vein. At various time points (0.5, 1, 2, 4, 8, 12, 24 hours), mice are euthanized, and organs (tumor, liver, spleen, kidney, heart, lung) and blood are collected. Tissues are imaged using an IVIS fluorescence imaging system, and fluorescence intensity is quantified. Alternatively, the distribution of labeled peptide is visualized by fluorescent microscopy of frozen tissue sections. For therapeutic studies, LyP-1 conjugated to a drug (e.g., doxorubicin) or toxin is administered IV at regular intervals (e.g., q3d or q7d for 2-4 weeks). Tumor volumes are measured with calipers, and survival is monitored. For atherosclerosis studies, 6-8 week old male ApoE-/- mice are fed a high-fat diet for 8-12 weeks to induce plaque formation. LyP-1 TFA is administered intraperitoneally (1-10 mg/kg) or intravenously (0.5-5 mg/kg) 3 times per week for 8 weeks. After treatment, the mice are euthanized, and the aorta is excised, fixed, and stained with Oil Red O for plaque area quantification. Cryosections of the aortic root are stained with H&E and for macrophages (Mac-2 or CD68). Apoptosis is assessed by TUNEL staining.
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| ADME/Pharmacokinetics |
LyP-1 TFA is a cyclic peptide (MW ~1106 Da). As a peptide, its pharmacokinetic properties are characterized by rapid renal clearance and proteolytic degradation in plasma, leading to a short half-life (typically <30 minutes for unmodified peptides). The cyclic structure (disulfide bridge) confers some stability against proteases compared to linear peptides, but the half-life is still short. The TFA salt improves solubility. The volume of distribution is limited to the vascular space and highly perfused tissues. The peptide is primarily eliminated by the kidneys. To improve pharmacokinetics for therapeutic applications, LyP-1 is often conjugated to nanoparticles, polymers (e.g., PEG), or encapsulated in liposomes, which prolongs circulation time and enhances tumor accumulation via the EPR effect. Detailed parameters (Tmax, Cmax, AUC, t½, CL, Vd) are not standardized and depend on the conjugate formulation.
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| Toxicity/Toxicokinetics |
LyP-1 TFA is a research tool and not a therapeutic agent. Toxicity data for the unmodified peptide is not published. In animal studies, intravenous or intraperitoneal administration of unconjugated LyP-1 peptide at doses up to 10 mg/kg appears to be well-tolerated in mice, with no overt signs of toxicity or significant weight loss. The primary toxicity would be related to the cargo (drug, toxin, nanoparticle) rather than the LyP-1 peptide itself. The TFA counterion may cause irritation. Standard safety precautions for handling peptides should be followed.
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| References |
[1]. Fan Zhang, et al.Imaging tumor-induced sentinel lymph node lymphangiogenesis with LyP-1 peptide. Amino Acids. 2012 Jun;42(6):2343-51.
[2]. Ningning Song, et al. Recent progress in LyP-1-based strategies for targeted imaging and therapy. Drug Deliv. 2019 Dec;26(1):363-375. |
| Additional Infomation |
LyP-1 (sequence: CGNKRTRGC) is a cyclic peptide discovered by phage display technology. Its target, p32 (gC1qR), is a multifunctional protein that is overexpressed in many solid tumors and in tumor-associated macrophages in hypoxic regions. The peptide is internalized by tumor cells upon binding, which is beneficial for intracellular drug delivery. LyP-1 has been used for tumor imaging, targeted drug delivery, and for studying tumor biology. It is strictly for research use and has not received regulatory approval for clinical use. The TFA salt form is used for improved solubility and stability.
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| Molecular Formula |
C38H66F3N17O14S2
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| Related CAS # |
LyP-1;454487-07-1
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| Appearance |
Typically exists as solid at room temperature
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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, avoid exposure to moisture. |
| 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 (~90.40 mM)
H2O :~100 mg/mL (~90.40 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.26 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 (2.26 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 (2.26 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
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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