Physicochemical Properties
| Molecular Formula | C28H26O10 |
| Molecular Weight | 522.500049114227 |
| Exact Mass | 582.173 |
| CAS # | 2130033-55-3 |
| PubChem CID | 145945112 |
| Appearance | Typically exists as solid at room temperature |
| LogP | 2.8 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 12 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 42 |
| Complexity | 993 |
| Defined Atom Stereocenter Count | 5 |
| SMILES | CC1(O[C@H]2[C@H](O[C@H]([C@@H]([C@H]2O1)O)OC3=C4COC(=O)C4=C(C5=CC(=C(C=C53)OC)OC)C6=CC7=C(C=C6)OCO7)CO)C |
| InChi Key | PMJUVXNBXQVOOP-NXMNBGQRSA-N |
| InChi Code | InChI=1S/C30H30O12/c1-30(2)41-26-21(10-31)39-29(24(32)27(26)42-30)40-25-15-9-19(35-4)18(34-3)8-14(15)22(23-16(25)11-36-28(23)33)13-5-6-17-20(7-13)38-12-37-17/h5-9,21,24,26-27,29,31-32H,10-12H2,1-4H3/t21-,24-,26+,27-,29+/m1/s1 |
| Chemical Name | 4-[[(3aS,4R,6S,7R,7aR)-7-hydroxy-4-(hydroxymethyl)-2,2-dimethyl-4,6,7,7a-tetrahydro-3aH-[1,3]dioxolo[4,5-c]pyran-6-yl]oxy]-9-(1,3-benzodioxol-5-yl)-6,7-dimethoxy-3H-benzo[f][2]benzofuran-1-one |
| 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 | ATP6V0A2, cellular apoptosis susceptibility (CAS)[2] |
| ln Vitro | PHY34 (0.001 nM–50 μM, 72 h) has the greatest effect on HGSOC cell lines and has a nanomolar influence on the proliferation of different cancer cells by inducing apoptosis based on elevated cPARP levels [1]. PHY34 inhibits autolysosomal breakdown in OVCAR8 and OVCAR3 at 100 nM and 1 μM, respectively, over a 24-hour period [1]. PHY34 (10 nM, 24 h) causes OVCAR8 to undergo apoptosis after inhibiting late autophagy [1]. PHY34 (100 nM, 48 h) causes OVCAR3 to undergo apoptosis after inhibiting late autophagy [1]. In H4 cells, PHY34 (0.01 nM-2 μM, 72 h) causes cell death when wild-type V0A2 is present, but not when the V823I mutant is present [2]. Several proteins in the nucleus have their subcellular localization altered by PHY34 (10, 100nM; 48 h, 72 h) [2]. PHY34 selectively binds to the ATP6V0A2 subunit (20 μM, 1 h) [2]. |
| ln Vivo | In the female nude mouse tumor-bearing model created with OVCAR8, PHY34 (0.75 mg/kg, ip, three times a week for three weeks) significantly suppressed tumor growth and decreased Ki67 expression in tumor tissues [1]. |
| Cell Assay |
Cell Viability Assay[1][2] Cell Types: OVCAR8, OVCAR3, HT-29, MDA-MB-435, MDA-MB -231, IOSE80, FT33 Tested Concentrations: 0.001 nM-50 μM Incubation Duration: 72 h Experimental Results: Inhibited the growth of various cancer cells with IC50 values of 4 nM(OVCAR8, OVCAR3), 43.3 nM(HT-29), 23 nM( MDA-MB-435), 5.2 nM(MDA-MB-231). demonstrated no toxicity to IOSE80 and FT33 (IC50 >50 μM). Cell Viability Assay[2] Cell Types: H4 Tested Concentrations: 0.01 nM-2 μM Incubation Duration: 72 h Experimental Results: Inhibited mutant cell with an IC50 value of 246 pM that was 1000-fold more potent than HTP-013(434 nM). Conferred resistance in V8231 mutation and no impact activity in T216A mutation. Apoptosis Analysis[1] Cell Types: OVCAR8, OVCAR3 Tested Concentrations: 10 nM, 100 nM Incubation Duration: 72 h Experimental Results: Increased the number of cells in early and late apoptosis in OVCAR8 and OVCAR3 at 10 nM and 100 nM, respectively. Cell Autophagy Assay[1] Cell Types: Hela Tested Concentrations: 9.31 fM-20 μM Incubation Duration: 4 h Experimental Results: Sustained high levels of LC3B puncta with an EC50 value of 2 nM. Inhibited autophagy with an EC50 value of 3.9 nM. Cell Autophagy Assay[2] Cell Types: OVCAR3 Tested Concentrations: 5 nM Incubation Duration: 4 h Experimental Results: Inhibited autophagy with an ED50 value of 6.29 nM, and was more potent than bafilomycin A1 (HY-100558) with an ED50 value of 29.1 nM. Western Blot Analysis[1][2] Cell Types: OVCAR8, OVCAR3, OVCAR4 Tested Concentrations: 10 nM, 100 nM Incubation Duration: 48 h, 72 h Experimental Results: Increased cPARP levels in OVCAR8 after 48 h at 10 nM, in OVCAR4 and OVCAR3 after 72 h at 100 nM, respectively. Reversed the conversion of PARP to cPARP combined with RAP (HY-10219) of 1 μM. Western Blot Analysis[2] Cell Types: OVCAR8, OVCAR3, OVCAR4 Tested Concentrations: 10 nM Incubation Duration: 24 h, 48 h, 72 h Experimental Results: Promoted histone H3, LAMP1/2, ACSS2, and PCNA nuclear protein accumulation at 48 h. decreased expression of KPNA2 (Karyopherin subunit alpha 2) with time-dependent manner. Increased nuclear accumulation of mutant p53 at 48 h. |
| Animal Protocol |
Animal/Disease Models: OVCAR8-induced xenograft models in female nude mice[1]. Doses: 0.75 mg/kg, three times a week for three weeks Route of Administration: intraperitoneal (ip)injection Experimental Results: diminished tumor burden based on average abdominal radiant efficiency with no gross toxicity through analysis of fluorescence imaging. |
| References |
[1]. Young AN,et al. Phyllanthusmin Derivatives Induce Apoptosis and Reduce Tumor Burden in High-Grade Serous Ovarian Cancer by Late-Stage Autophagy Inhibition. Mol Cancer Ther. 2018 Oct;17(10):2123-2135. [2]. Salvi A, et al. PHY34 inhibits autophagy through V-ATPase V0A2 subunit inhibition and CAS/CSE1L nuclear cargo trafficking in high grade serous ovarian cancer. Cell Death Dis. 2022 Jan 10;13(1):45. |
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 | 1.9139 mL | 9.5694 mL | 19.1388 mL | |
| 5 mM | 0.3828 mL | 1.9139 mL | 3.8278 mL | |
| 10 mM | 0.1914 mL | 0.9569 mL | 1.9139 mL |