TC6-D3
Based on 1 Customer Validation
TC6-D3 is a proteolysis-resistant peptide and also a CC chemokine receptor 7 (CCR7) inhibitor, with a Kd value of 403 nM against mouse targets. TC6-D3 blocks the interaction between CCR7 and its ligands CCL19 and CCL21, and inhibits the activation of the ERK1/2 pathway. TC6-D3 reduces the migratory capacity of tumor cells in vitro. TC6-D3 inhibits tumor growth and lymph node metastasis in vivo. TC6-D3 restores T cell cytotoxicity, promotes CD8+ T cell infiltration, and enhances anti-tumor immune responses. TC6-D3 can be used in studies related to lymph node metastasis.
For research use only. We do not sell to patients.
- Purity : 98.55%
- CAS No.: 3121532-43-9
- Formula: C61H98N12O19
- Molecular Weight:1303.50
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Storage:
Sealed storage, away from moisture.
Powder -80°C, 2 years , -20°C, 1 year* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Biological Activity
Description
IC50 & Target
[1]|
CCR7 403 nM (Kd) |
ERK1 |
ERK2 |
In Vitro
TC6-D3 peptide (0.2 mM; 0.2548 h) exhibits significantly enhanced proteolysis resistance compared to TC6, retaining ~70% of its initial concentration after 48 h incubation in 10% mouse serum at 37°C[1].
TC6-D3 peptide (5 min) binds specifically to mouse CCR7 with a dissociation constant of 403 nM, as measured via MST with CHOK1-mCCR7-EGFP cell membrane proteins[1].
TC6-D3 peptide (12.5-200 μM; 24-72 h) does not inhibit the proliferation of MC38-CCR7 (GFP) cells at concentrations ranging from 12.5 μM to 200 μM over 24 h, 48 h, or 72 h incubations[1].
TC6-D3 peptide (200 μM; 48 h) significantly inhibits CCL19-induced chemotaxis of MC38-CCR7 (GFP) cells in a 48 h transwell assay[1].
TC6-D3 peptide (200 μM) significantly attenuates CCL19- and CCL21-induced ERK1/2 phosphorylation in MC38-CCR7 (GFP) and B16-CCR7 cells[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:MC38-CCR7 (GFP) cells
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Concentration:12.5 μM; 25 μM; 50 μM; 100 μM; 200 μM
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Incubation Time:24 h, 48 h, 72 h
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Result:Showed no significant inhibitory effect on the proliferation of MC38-CCR7 (GFP) cells across all tested concentrations and incubation times.
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Cell Line:MC38-CCR7 (GFP) and MC38-V (GFP) cells
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Concentration:200 μM
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Incubation Time:48 h
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Result:Significantly inhibited CCL19-induced migration of MC38-CCR7 (GFP) cells, while CCL19 had only a weak effect on MC38-V (GFP) cell migration.
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Cell Line:MC38-CCR7 (GFP) and B16-CCR7 cells
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Concentration:200 μM
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Incubation Time:5 min; 10 min; 20 min
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Result:Significantly inhibited CCL19- or CCL21-induced increase in phospho-ERK1/2 levels in both MC38-CCR7 (GFP) and B16-CCR7 cells.
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:C57BL/6 (female, 6 to 8 weeks old, MC38-CCR7 (GFP) cell footpad injection-induced popliteal lymph node metastasis model)[1]
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Dosage:2 mg/kg; 6 mg/kg
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Administration:i.p.; daily; 14 days
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Result:Slowed primary tumor growth at 2 mg/kg, with more prominent anti-tumor activity observed at 6 mg/kg; tumor weight followed the same trend.
Significantly inhibited popliteal lymph node metastasis at 6 mg/kg, as measured by IVIS imaging and reduced frequencies of CCR7+GFP+ tumor cells in popliteal lymph nodes via flow cytometry; no significant effect on inguinal lymph node metastasis was observed at either dose.
Showed no significant change in body weight compared to controls.
Significantly increased the frequency of intratumoral CD8+ T cells to 35.3% of CD45+ cells (vs. 12.1% in controls) and intratumoral IFN-γ-secreting CD8+ T cells to 9.66% (vs. 4.73% in controls) at 6 mg/kg; caused a smaller increase in these populations at 2 mg/kg.
Increased the frequency of IFN-γ-secreting CD8+ T cells in popliteal lymph nodes to 2.37% (vs. 0.92% in controls) and inguinal lymph nodes to 3.94% (vs. 2.26% in controls) at 6 mg/kg; both doses showed this effect.
Caused no significant changes in the frequency of total DCs or CCR7+ DCs in primary tumors, popliteal lymph nodes, or inguinal lymph nodes.
Maintained serum ALT, AST, BUN, and CREA levels within normal ranges, and showed no abnormalities in histopathology of major organs.
- Cell Cytotoxicity Assay
- Mammalian live/dead viability and cytotoxicity staining
- Cell migration
- Patient-Derived Xenograft (PDX)
- Patient-Derived Orthotopic Xenograft (PDOX)
- Subcutaneous Cell-Line-Derived Xenograft
- Orthotopic Cell-Line Xenograft
- Research Protocol for Cancer Immunology
- Cell Viability Determination by MTT Colorimetric Assay
Chemical Information
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CAS No. 3121532-43-9
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Appearance Solid
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Molecular Weight 1303.50
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Formula C61H98N12O19
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Sequence
d{Leu-Ser-Pro}-Leu-Ile-Phe-Val-Thr-Thr-Pro-Asp-{d-Thr}
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Sequence Shortening
d{Leu-Ser-Pro}-LIFVTTPD-{d-Thr}
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Sealed storage, away from moisture
Powder -80°C 2 years -20°C 1 year * In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Solvent & Solubility
In Vitro:
DMSO : 50 mg/mL (38.36 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: 2.5 mg/mL (1.92 mM); Clear solution; Need ultrasonic
This protocol yields a clear solution of 2.5 mg/mL.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (1.92 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL. * In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 0.7672 mL | 3.8358 mL | 7.6717 mL | 19.1791 mL |
| 5 mM | 0.1534 mL | 0.7672 mL | 1.5343 mL | 3.8358 mL | |
| 10 mM | 0.0767 mL | 0.3836 mL | 0.7672 mL | 1.9179 mL | |
| 15 mM | 0.0511 mL | 0.2557 mL | 0.5114 mL | 1.2786 mL | |
| 20 mM | 0.0384 mL | 0.1918 mL | 0.3836 mL | 0.9590 mL | |
| 25 mM | 0.0307 mL | 0.1534 mL | 0.3069 mL | 0.7672 mL | |
| 30 mM | 0.0256 mL | 0.1279 mL | 0.2557 mL | 0.6393 mL |