Physicochemical Properties
| Molecular Formula | C228H388N86O64 |
| Molecular Weight | 5358.06 |
| Exact Mass | 4520.814 |
| CAS # | 2460055-10-9 |
| Related CAS # | FOXO4-DRI acetate |
| Appearance | White to off-white solid powder |
| InChi Key | PFXXSSUGMGVDLB-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C31H32Cl2N4O2.C30H33ClN4O2.C30H34N4O3.C30H34N4O2.C28H34Cl2N4O2.C26H31BrFN5O2.C26H30BrFN4O2.C26H30Cl2N4O2.C22H20N4O/c32-27-14-16-30(33)35-29(27)21-25(22-34)31(38)36-28(15-9-23-7-3-1-4-8-23)24-10-12-26(13-11-24)39-20-19-37-17-5-2-6-18-37;1-2-7-28(22-11-14-26(15-12-22)37-19-18-35-16-4-3-5-17-35)34-30(36)24(21-32)20-25-13-10-23-8-6-9-27(31)29(23)33-25;1-2-7-27(22-11-14-26(15-12-22)37-19-18-34-16-4-3-5-17-34)33-30(36)24(21-31)20-25-13-10-23-8-6-9-28(35)29(23)32-25;1-2-8-28(24-12-15-27(16-13-24)36-20-19-34-17-6-3-7-18-34)33-30(35)25(22-31)21-26-14-11-23-9-4-5-10-29(23)32-26;1-20(2)6-12-25(33-28(35)22(19-31)18-26-24(29)11-13-27(30)32-26)21-7-9-23(10-8-21)36-17-16-34-14-4-3-5-15-34;1-3-4-23(31-26(34)20(18-29)17-24-22(28)9-10-25(27)30-24)19-5-7-21(8-6-19)35-16-15-33-13-11-32(2)12-14-33;1-2-6-23(31-26(33)20(18-29)17-24-22(28)11-12-25(27)30-24)19-7-9-21(10-8-19)34-16-15-32-13-4-3-5-14-32;1-2-6-23(31-26(33)20(18-29)17-24-22(27)11-12-25(28)30-24)19-7-9-21(10-8-19)34-16-15-32-13-4-3-5-14-32;1-2-7-20(16-8-4-3-5-9-16)26-22(27)18(15-23)14-19-12-11-17-10-6-13-24-21(17)25-19/h1,3-4,7-8,10-14,16,21,28H,2,5-6,9,15,17-20H2,(H,36,38);6,8-15,20,28H,2-5,7,16-19H2,1H3,(H,34,36);6,8-15,20,27,35H,2-5,7,16-19H2,1H3,(H,33,36);4-5,9-16,21,28H,2-3,6-8,17-20H2,1H3,(H,33,35);7-11,13,18,20,25H,3-6,12,14-17H2,1-2H3,(H,33,35);5-10,17,23H,3-4,11-16H2,1-2H3,(H,31,34);2*7-12,17,23H,2-6,13-16H2,1H3,(H,31,33);3-6,8-14,20H,2,7H2,1H3,(H,26,27) |
| Chemical Name | 3-(6-bromo-3-fluoropyridin-2-yl)-2-cyano-N-[1-[4-[2-(4-methylpiperazin-1-yl)ethoxy]phenyl]butyl]prop-2-enamide;3-(6-bromo-3-fluoropyridin-2-yl)-2-cyano-N-[1-[4-(2-piperidin-1-ylethoxy)phenyl]butyl]prop-2-enamide;3-(8-chloroquinolin-2-yl)-2-cyano-N-[1-[4-(2-piperidin-1-ylethoxy)phenyl]butyl]prop-2-enamide;2-cyano-3-(3,6-dichloropyridin-2-yl)-N-[4-methyl-1-[4-(2-piperidin-1-ylethoxy)phenyl]pentyl]prop-2-enamide;2-cyano-3-(3,6-dichloropyridin-2-yl)-N-[3-phenyl-1-[4-(2-piperidin-1-ylethoxy)phenyl]propyl]prop-2-enamide;2-cyano-3-(3,6-dichloropyridin-2-yl)-N-[1-[4-(2-piperidin-1-ylethoxy)phenyl]butyl]prop-2-enamide;2-cyano-3-(8-hydroxyquinolin-2-yl)-N-[1-[4-(2-piperidin-1-ylethoxy)phenyl]butyl]prop-2-enamide;2-cyano-3-(1,8-naphthyridin-2-yl)-N-(1-phenylbutyl)prop-2-enamide;2-cyano-N-[1-[4-(2-piperidin-1-ylethoxy)phenyl]butyl]-3-quinolin-2-ylprop-2-enamide |
| 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 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) |
Biological Activity
| ln Vitro | In senescent TM3 Leydig cells, FOXO4-DRI (25 mM; 3 days) induces apoptosis and causes nuclear exclusion of active p53 [1]. The senescence level of PDL9 cells is significantly reduced by FOXO4-DRI (25 μM; 5 days) [2]. |
| ln Vivo | In naturally aging rats, FOXO4-DRI (5 mg/kg; ip; every other day for three administrations) enhances the testicular microenvironment and relieves inadequate testosterone secretion [1]. |
| Cell Assay |
Cell Viability Assay[1] Cell Types: Senescent Leydig cells Tested Concentrations: 25 mM Incubation Duration: 3 days Experimental Results: decreased the viability of senescent as compared to normal TM3 Leydig cells. Apoptosis Analysis[1] Cell Types: Senescent Leydig cells Tested Concentrations: 25 mM Incubation Duration: 3 days Experimental Results: The apoptosis rate increased from 10% to 27%. Western Blot Analysis[2] Cell Types: PDL9 cells Tested Concentrations: 25 μM Incubation Duration: 5 days Experimental Results: diminished the protein levels of representative senescent markers, including p16, p21, and p53. RT-PCR[2] Cell Types: PDL9 cells Tested Concentrations: 25 μM Incubation Duration: 5 days Experimental Results: Enhanced SOX9 expression, and decreased MMP12 and MMP13 expression. |
| Animal Protocol |
Animal/Disease Models: Naturally aged male C57BL/6 mice (20-24 months old)[1] Doses: 5 mg/kg Route of Administration: intraperitoneal (ip)injection, every other day for three administrations Experimental Results: Increased serum testosterone levels. Increased levels of both 3β-HSD and CYP11A1. diminished interstitial SA-β-gal activity and lowered levels of senescence-associated proteins p53, p21, and p16. diminished the levels of IL- 1β, IL-6 and TGF-β. |
| References |
[1]. FOXO4-DRI alleviates age-related testosterone secretion insufficiency by targeting senescent Leydig cells in aged mice. Aging (Albany NY). 2020 Jan 20;12(2):1272-1284. [2]. Senolytic Peptide FOXO4-DRI Selectively Removes Senescent Cells From in vitro Expanded Human Chondrocytes. Front Bioeng Biotechnol. 2021 Apr 29;9:677576. |
Solubility Data
| Solubility (In Vitro) | DMSO : 50 mg/mL (9.33 mM) |
| 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 | 0.1866 mL | 0.9332 mL | 1.8663 mL | |
| 5 mM | 0.0373 mL | 0.1866 mL | 0.3733 mL | |
| 10 mM | 0.0187 mL | 0.0933 mL | 0.1866 mL |