| ln Vitro |
NCP26 is an ATP-competitive recombinant ProRS inhibitor. Its binding affinity increases more than 750-fold in the presence of 100 μM proline, with a KD value of 0.35 nM[1]. NCP26 (0.5 μM; 48 h) maintains antiproliferative activity against AMO1 and RPMI 8226 multiple myeloma (MM) cells in the range of 0 to 20 mM proline concentrations, and its potency is not reduced at high proline concentrations[1]. NCP26 (0.01–10 μM; 96 h) is 10-fold more potent against AMO1 multiple myeloma (MM) cells than against healthy donor peripheral blood mononuclear cells (PBMCs), with an EC50 value of 0.1 μM for the former and approximately 1 μM for the latter after 96 h of treatment[1]. NCP26 can effectively inhibit the growth of most multiple myeloma (MM) cell lines, with EC50 values ranging from 135 nM to 1.1 μM[1]. NCP26 (0.5 μM; 6–48 h) can induce G0/G1 phase cell cycle arrest and caspase-dependent apoptosis in AMO1 and RPMI 8226 multiple myeloma (MM) cells[1].
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| ln Vivo |
NCP26 (2.5–10 mg/kg; intraperitoneal injection; once daily; for 21 days) significantly inhibited the growth of multiple myeloma tumors in CB17 SCID mice carrying AMO1 xenografts (P = 0.01 and P < 0.001, respectively) and prolonged their overall survival (P = 0.01 and P < 0.001, respectively), without causing significant weight loss [1]. NCP26 (10 mg/kg; once daily) also significantly inhibited the growth of multiple myeloma tumors in AMO1 xenograft mouse models (P = 0.005) and prolonged host survival (P < 0.001), without causing weight loss [2].
|
| Cell Assay |
Cell cycle analysis [1]
Cell Types: AMO1, RPMI 8226 MM cell lines Tested Concentrations: 0.5 μM Incubation Duration: 6 hours, 24 hours, 48 hours Experimental Results: At 6 hours, both cell lines induced G0/G1 phase cell cycle arrest, and the degree of arrest increased at 24 hours. Induced apoptosis, manifested as an increase in Annexin-V positive cells, sub-G1 phase cell populations, and mitochondrial damage. Induced cleavage of caspase-3, -8, -9, and PARP, with cleavage in a time- and dose-dependent manner. Western Blot Analysis [1] Cell Types: AMO1, RPMI 8226 MM cell lines Tested Concentrations: 0-1 μM Incubation Duration: 6 hours Experimental Results: The protein levels of MYC, PIM2, CCND1, and TCF3 in both cell lines decreased in a dose-dependent manner. This is consistent with the effect of EPRS shRNA knockdown on the protein levels of MYC, PIM2, CCND1, and TCF3. |
| Animal Protocol |
Animal/Disease Models:CB17 SCID (5-week-old females, subcutaneously inoculated with AMO1 cells)[1]
Doses: 2.5 mg/kg; 10 mg/kg Route of Administration: Intraperitoneal injection; once daily for 21 days Experimental Results: Compared with the vector control group, AMO1 tumor growth was significantly inhibited (control group vs. 2.5 mg/kg, P = 0.01; control group vs. 10 mg/kg, P < 0.001). The tumor growth inhibition effect of the 10 mg/kg dose group was better than that of the 2.5 mg/kg dose group. No significant weight loss was observed in either treatment group. Overall survival was significantly prolonged compared with the vector control group (control group vs. 2.5 mg/kg, P = 0.01; control group vs. 10 mg/kg, P < 0.001). In the treatment group, the expression of MYC, CCND1, and TCF3 was downregulated, while the expression of p-GCN2 and DDIT3 was upregulated in the tumors. |
| References |
| Molecular Formula |
C20H23N5O2
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|---|---|
| Molecular Weight |
365.44
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| CAS # |
2396683-89-7
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| Appearance |
Typically exists as solids at room temperature
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| SMILES |
O=C(NC1CC=2C=CC=CC2C1)C3=NC=CN=C3NC(=O)N4CCCCC4
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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)
|
| 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
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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 | 2.7364 mL | 13.6821 mL | 27.3643 mL | |
| 5 mM | 0.5473 mL | 2.7364 mL | 5.4729 mL | |
| 10 mM | 0.2736 mL | 1.3682 mL | 2.7364 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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