API-1 (API1) is a novel and potent Akt inhibitor that triggers Mcl-1 degradation that is GSK3-dependent, β-TrCP- and FBXW7-mediated, leading to the induction of apoptosis. With an IC50 of about 0.8 M, it effectively lowers the levels of Akt phosphorylation. Specifically in API-1-sensitive lung cancer cell lines, API-1 quickly and effectively decreased the levels of Mcl-1. The ectopic expression of Mcl-1 prevented API-1 from causing apoptosis in cells. The proteasome was inhibited with MG132 to reverse the reduction in Mcl-1 half-life caused by API-1 treatment. Mcl-1 phosphorylation (S159/T163) increased quickly after API-1 reduced its levels. Furthermore, GSK3 inhibition blocked API-1-induced Mcl-1 phosphorylation and reduction and counteracted API-1's ability to induce apoptosis.
Physicochemical Properties
| Molecular Formula | C13H15N5O6 |
| Molecular Weight | 337.2881 |
| Exact Mass | 337.102 |
| Elemental Analysis | C, 46.29; H, 4.48; N, 20.76; O, 28.46 |
| CAS # | 36707-00-3 |
| Related CAS # | 36707-00-3 |
| PubChem CID | 24773090 |
| Appearance | white solid powder |
| Density | 1.792g/cm3 |
| Boiling Point | 795.8ºC at 760 mmHg |
| Flash Point | 435.1ºC |
| Index of Refraction | 1.764 |
| LogP | -1.9 |
| Hydrogen Bond Donor Count | 5 |
| Hydrogen Bond Acceptor Count | 10 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 24 |
| Complexity | 571 |
| Defined Atom Stereocenter Count | 4 |
| SMILES | NC(C1C(=O)C2=C(N=CN=C2N)N(C2C(O)C(O)C(CO)O2)C=1)=O |
| InChi Key | SPBWHPXCWJLQRU-FITJORAGSA-N |
| InChi Code | InChI=1S/C13H15N5O6/c14-10-6-7(20)4(11(15)23)1-18(12(6)17-3-16-10)13-9(22)8(21)5(2-19)24-13/h1,3,5,8-9,13,19,21-22H,2H2,(H2,15,23)(H2,14,16,17)/t5-,8-,9-,13-/m1/s1 |
| Chemical Name | 4-amino-8-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-5-oxopyrido[2,3-d]pyrimidine-6-carboxamide |
| Synonyms | API1; NSC 177233;API-1; NSC-177233; api-1; 36707-00-3; NSC 177223; 4-Amino-5,6,7,8-tetrahydro-5-oxo-8-(beta-D-ribofuranosyl)pyrido[2,3-d]pyrimidine-6-carboxamide; W3UEK36Q7X; NSC177223; 4-amino-5,8-dihydro-5-oxo-8-beta-d-ribofuranosyl-pyrido[2,3-d]pyrimidine-6-carboxamide; ld-101; API 1; NSC177233 |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets | Akt |
| ln Vitro | An isosteric scaffold of adenosine, the 4-aminopyrido[2,3-d]pyrimidine derivative API-1 was discovered to inhibit Akt by screening the DTP/NCI compound library in a cell-based assay. API-1 binds to the PH domain and inhibits Akt membrane translocation, leading to the inhibition of the growth of tumors with hyperactivated Akt. API-1 was shown to induce apoptosis in tested cancer cell lines by synergizing with TNF-related apoptosis-inducing ligand (TRAIL). A series of 4-amino-pyrrolo[2,3-d]pyrimidine derivatives, closely related to API-1, were discovered in a high-throughput screening based on a newly developed fluorescence-based assay as allosteric Akt inhibitors [1]. |
| References |
[1]. Akt inhibitors in cancer treatment: The long journey from drug discovery to clinical use (Review). Int J Oncol . 2016 Mar;48(3):869-85. [2]. A small molecule inhibits Akt through direct binding to Akt and preventing Akt membrane translocation. J Biol Chem. 2016 Oct 21;291(43):22856. |
| Additional Infomation | Screening of the National Cancer Institute (NCI) Diversity Set led to the identification of API-2 [(Triciribine (TCN), NSC 154020] that potently inhibited Akt signaling in human cancer cells, leading to the inhibition of cell growth and the induction of apoptosis. TCN is a tricyclic purine nucleoside derivative (Fig. 9) that is metabolically activated inside cells by adenosine kinase to its monophosphate active analog, TCN-P. Surface plasmon resonance and nuclear magnetic resonance spectroscopy have demonstrated that TCN-P, but not TCN, binds to the PH domain of Akt in the vicinity of the PIP3 binding pocket, thereby preventing Akt phosphorylation and subsequent activation. The proposed mechanism of TCN-P Akt inhibition is by preventing PIP3 from recruiting Akt to the plasma membrane, either by competing with PIP3 for binding to PH domain or by binding to region that induces conformational changes that hinder PIP3 activation. Treatment of T cell acute lymphocytic leukemia cells with 10 μM TCN has been shown to inhibit Akt phosphorylation and its downstream signaling, causing cell cycle arrest and caspase-dependent apoptosis. In prostate cancer cells, TCN was shown to increase the apoptosis induced by the death receptor pathway (124). TCN and TCN-P were subjected to several clinical trials against various solid neoplasms and hematological malignancies, but their efficacy is limited due to their toxicity. The combination treatment of TCN with other anticancer agents proved to be a better solution than treatment with TCN alone[1]. |
Solubility Data
| 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 |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9648 mL | 14.8240 mL | 29.6481 mL | |
| 5 mM | 0.5930 mL | 2.9648 mL | 5.9296 mL | |
| 10 mM | 0.2965 mL | 1.4824 mL | 2.9648 mL |