| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Targets |
Alkaline phosphatase (alkaline phosphatase-dependent mechanism). D-3 is a phosphorpeptide that selectively eliminates iPSCs and ESCs. It is not a classical enzyme inhibitor but a cytotoxic agent that targets pluripotent stem cells through an alkaline phosphatase-dependent mechanism.
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| ln Vitro |
D-3 inhibits the generation of residual teratomas produced by iPSCs in a mouse tumorigenicity assay[1]. With a half maximum inhibitory concentration value of 192.3 ± 57.4 μM, D-3 clearly causes 201B7 cells to lose viability[1]. an elevation of the quantity of p38, p44/42, and elF2a MAPK that are activated in response to cellular stress. D-3 administration also results in an increase in Annexin-V and SYTOX-positive cells, which indicates apoptotic and dead cells[1]. D-3 has minimal effect on a variety of non-iPSCs, such as neurons and hepatocytes[1].
D-3 induces toxicity in cultured iPSCs and ESCs after 1 hour of incubation via an alkaline phosphatase-dependent mechanism. It clearly causes 201B7 iPSC cells to lose viability with a half-maximum inhibitory concentration (IC50) value of 192.3 +/- 57.4 microM. It does not affect differentiated cells, making it selective for pluripotent stem cells. |
| ln Vivo |
In vivo, D-3 effectively inhibits the formation of teratomas by residual induced pluripotent stem cells (iPSCs) in a mouse tumorigenicity assay. By selectively eliminating residual pluripotent cells that may form teratomas after transplantation, D-3 enhances the safety of iPSC-based cell therapies. This makes it a valuable tool for regenerative medicine research.
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| Enzyme Assay |
D-3 is not evaluated in standard cell-free enzyme assays because its mechanism involves alkaline phosphatase-dependent cytotoxicity rather than direct enzyme inhibition. Instead, its binding to alkaline phosphatase can be assessed using surface plasmon resonance (SPR) or by measuring alkaline phosphatase activity in the presence of D-3 using a fluorogenic substrate (e.g., 4-methylumbelliferyl phosphate).
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: Six human iPSCs lines and one human ESC line (khES1). Tested Concentrations: 400 μM. Incubation Duration: 1-2 h. Experimental Results: Sufficient to induce a viability loss (>99% in all iPSC lines and >95% in khES1). For cell-based assays, iPSCs (e.g., 201B7 cells) or ESCs are seeded in 96-well plates and treated with D-3 at varying concentrations (0-500 uM) for 1-24 hours. Cell viability is assessed by MTT, CCK-8, or ATP-based luminescence assays. The IC50 for 201B7 cells is 192.3 +/- 57.4 microM. Selectivity is confirmed by testing differentiated cells (e.g., fibroblasts) in parallel. Teratoma formation can be assessed in soft agar colony formation assays. |
| Animal Protocol |
For animal studies, D-3 is co-injected or administered after iPSC transplantation in a mouse tumorigenicity model. iPSCs (1 x 106 cells) are mixed with D-3 (typically 200-500 uM) and injected subcutaneously into immunodeficient mice. Teratoma formation is monitored over 8-12 weeks. The number, size, and weight of teratomas are measured. D-3 treatment should significantly reduce or eliminate teratoma formation compared to control (iPSCs alone).
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| ADME/Pharmacokinetics |
D-3 is a phosphorpeptide that is typically dissolved in DMSO or PBS for in vitro use. For in vivo studies, it is co-injected with iPSCs in PBS. The compound has a molecular weight of 870.88 and a molecular formula of C48H47N4O10P. Detailed pharmacokinetic parameters (half-life, absorption, distribution) have not been extensively reported.
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| Toxicity/Toxicokinetics |
D-3 is used in research to eliminate iPSCs and prevent teratoma formation. At the doses used in cell culture (up to 500 uM), it is selectively toxic to pluripotent stem cells but does not affect differentiated cells. In animal studies, no significant systemic toxicity has been reported at the doses used for teratoma prevention. Standard safety precautions for handling research chemicals should be followed.
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| References | |
| Additional Infomation |
D-3 (CAS: 1967815-98-0) is a phosphorpeptide that selectively eliminates iPSCs and ESCs via an alkaline phosphatase-dependent mechanism. It is a valuable tool for enhancing the safety of stem cell-based therapies by preventing teratoma formation from residual pluripotent cells. D-3 is not an approved drug and is intended for research applications only.
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| Molecular Formula |
C48H47N4O10P
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|---|---|
| Molecular Weight |
870.881353616714
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| Exact Mass |
870.302
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| CAS # |
1967815-98-0
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| PubChem CID |
138991764
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| Appearance |
White to off-white solid powder
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| LogP |
6.3
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| Hydrogen Bond Donor Count |
7
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
20
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| Heavy Atom Count |
63
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| Complexity |
1530
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| Defined Atom Stereocenter Count |
4
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| SMILES |
C(C1C=CC2C=CC=CC=2C=1)C(=O)N[C@H](CC1C=CC=CC=1)C(=O)N[C@H](CC1C=CC=CC=1)C(=O)N[C@@H](C(=O)N[C@@H](C(=O)O)CC1C=CC(OP(O)(O)=O)=CC=1)CC1C=CC=CC=1
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| InChi Key |
VSVONENCEIBGNL-MRWFHJSOSA-N
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| InChi Code |
InChI=1S/C48H47N4O10P/c53-44(31-36-20-23-37-18-10-11-19-38(37)26-36)49-40(27-32-12-4-1-5-13-32)45(54)50-41(28-33-14-6-2-7-15-33)46(55)51-42(29-34-16-8-3-9-17-34)47(56)52-43(48(57)58)30-35-21-24-39(25-22-35)62-63(59,60)61/h1-26,40-43H,27-31H2,(H,49,53)(H,50,54)(H,51,55)(H,52,56)(H,57,58)(H2,59,60,61)/t40-,41-,42-,43-/m1/s1
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| Chemical Name |
(2R)-2-[[(2R)-2-[[(2R)-2-[[(2R)-2-[(2-naphthalen-2-ylacetyl)amino]-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-3-(4-phosphonooxyphenyl)propanoic acid
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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: ≥ 53.3 mg/mL (61.20 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 | 1.1483 mL | 5.7413 mL | 11.4826 mL | |
| 5 mM | 0.2297 mL | 1.1483 mL | 2.2965 mL | |
| 10 mM | 0.1148 mL | 0.5741 mL | 1.1483 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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