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Picropodophyllin (also known as Picropodophyllotoxin, AXL1717 or PPP), a naturally occuring cyclolignan alkaloid isolated from the mayapple plant family, is a novel, potent, orally bioavailable and selective small molecule inhibitor of the IGF-1R with potential antineoplastic activity. In SCID mice xenografted with human ES-1, BE, and PC3, it exhibits strong in vivo antitumor efficacy.
Physicochemical Properties
| Molecular Formula | C22H22O8 |
| Molecular Weight | 414.41 |
| Exact Mass | 414.131 |
| Elemental Analysis | C, 63.76; H, 5.35; O, 30.89 |
| CAS # | 477-47-4 |
| Related CAS # | Picropodophyllin-d6 |
| PubChem CID | 72435 |
| Appearance | white solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 597.9±50.0 °C at 760 mmHg |
| Melting Point | 225-227ºC |
| Flash Point | 210.2±23.6 °C |
| Vapour Pressure | 0.0±1.8 mmHg at 25°C |
| Index of Refraction | 1.606 |
| LogP | 1.6 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 30 |
| Complexity | 629 |
| Defined Atom Stereocenter Count | 4 |
| SMILES | O1C([C@@]2([H])[C@]([H])(C3C([H])=C(C(=C(C=3[H])OC([H])([H])[H])OC([H])([H])[H])OC([H])([H])[H])C3=C([H])C4=C(C([H])=C3[C@@]([H])([C@@]2([H])C1([H])[H])O[H])OC([H])([H])O4)=O |
| InChi Key | YJGVMLPVUAXIQN-HAEOHBJNSA-N |
| InChi Code | InChI=1S/C22H22O8/c1-25-16-4-10(5-17(26-2)21(16)27-3)18-11-6-14-15(30-9-29-14)7-12(11)20(23)13-8-28-22(24)19(13)18/h4-7,13,18-20,23H,8-9H2,1-3H3/t13-,18+,19+,20-/m0/s1 |
| Chemical Name | (5R,5aR,8aS,9R)-5-hydroxy-9-(3,4,5-trimethoxyphenyl)-5a,6,8a,9-tetrahydro-5H-[2]benzofuro[5,6-f][1,3]benzodioxol-8-one |
| Synonyms | Picropodophyllotoxin, AXL1717; PPP; AXL-1717; AXL 1717; PPP; picropodophyllin |
| 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 |
IGF-1R (IC50 = 1 nM) Insulin-like Growth Factor 1 Receptor (IGF-1R) (IC50 = 8.5 nM for recombinant human IGF-1R kinase); no activity against IR, EGFR, HER2 (IC50 > 1000 nM) [1] - Confirmed IGF-1R as primary target (multiple myeloma model; no additional IC50 values) [2][5] - Confirmed IGF-1R targeting (hepatocellular carcinoma model; consistent with [1]’s IC50) [4] |
| ln Vitro |
PPP effectively inhibits the phosphorylation of Erk1/2, Akt (Ser 473), and IGF-1-stimulated IGF-1R in intact cells. Picropodophyllin specifically suppresses growth and causes IGF-1R-positive tumor cells grown in culture to undergo apoptosis.[1] By further reducing cell viability and increasing apoptosis, picropodophyllin synergistically sensitizes HMCL, primary human MM, and murine 5T33MM cells to ABT-737 and ABT-199.[3] Sorafenib and picropodialdoxolin both work in concert to inhibit the growth and motility of hepatocellular carcinoma cells.[4] Inhibited solid tumor cell proliferation: Breast cancer MCF-7 (IC50 = 19.2 nM), lung cancer H460 (IC50 = 22.6 nM); 100 nM Picropodophyllin reduced MCF-7 colony formation by 75% (14-day culture) [1] - Suppressed multiple myeloma (MM) cell growth: 5T33MM.1 (IC50 = 15.8 nM), RPMI-8226 (IC50 = 18.3 nM); 50 nM Picropodophyllin decreased p-IGF-1R (Tyr1135/1136) by 90% in 5T33MM.1 cells (2 hours) [2][5] - Induced MM cell apoptosis: 200 nM Picropodophyllin increased Annexin V-positive RPMI-8226 cells from 7% to 45% (48 hours); G2/M phase accumulation (from 18% to 42%, flow cytometry) [5] - Synergized with BH3-mimetic (ABT-737) in MM: 100 nM Picropodophyllin + 50 nM ABT-737 reduced viability by 82% (vs. 45%/38% for monotherapy); synergistic index = 0.56 [3] - Synergized with sorafenib in(HCC): 150 nM Picropodophyllin + 200 nM sorafenib reduced HepG2 viability by 78% (vs. 42%/35% for monotherapy); inhibited migration by 68% [4] |
| ln Vivo |
Picropodophyllin (20 mg/kg/12 h, i.p.) completely suppresses tumor growth in SCID mice xenografted with human ES-1, BE, and PC3.[1] Picropodophyllin also significantly increases survival in the 5T33MM mouse model and exhibits strong antitumor activity.[2] In 5T33MM multiple myeloma mice (C57BL/KaLwRij): Intraperitoneal Picropodophyllin (10 mg/kg, twice daily) for 21 days reduced tumor burden by 72%; median survival extended from 35 days (vehicle) to 62 days [2] - In nude mice bearing RPMI-8226 xenografts: Oral Picropodophyllin (15 mg/kg/day) for 28 days achieved 76% tumor growth inhibition (TGI); tumor p-IGF-1R reduced by 78% [5] - In nude mice bearing HepG2 HCC xenografts: Picropodophyllin (12 mg/kg/day, oral) + sorafenib (30 mg/kg/day, oral) for 35 days reduced tumor volume by 83% (vs. 45%/40% for monotherapy) [4] |
| Enzyme Assay |
The phosphorylation of pTG by IGF-1R-catalyzed substrate is assayed using a 96-well plate tyrosine kinase assay kit. To represent "non-IGF-1R tyrosine kinases," we use recombinant epidermal growth factor receptor, immunoprecipitated IR from HEPG2, immunoprecipitated IGF-1R from P6 cells, and IGF-1R immunodepleted supernatant from P6. The kinase reaction is triggered by the addition of ATP after the receptors have been treated for 30 minutes with the desired compounds in the kinase buffer (20 mM HEPES buffer (pH 7.4), 0.1 MnCl2, 0.2 Na3VO4, and 20 mM MgCl2). A phosphotyrosine-specific monoclonal antibody conjugated to horseradish peroxidase, clone PT-66, is used to probe the phosphorylated polymer substrate. O-phenylenediamine dihydrochloride, a chromogenic substrate for horseradish peroxidase, is used to develop color, and spectrophotometry (an ELISA reader) enables quantification. An ELISA sandwich test is used to measure IGF-1R tyrosine autophosphorylation. IGF-1R β-subunit antibody (1 μg/well) is coated onto 96-well plates and allowed to sit overnight at 4°C. 80 μg/well of the P6 cell line's total protein lysate is added after the plates are blocked for one hour with 1% BSA in PBS Tween. Total R-cell line protein lysate is used as a negative control. Before kinase activation with ATP, the evaluated compounds are added to tyrosine kinase buffer at room temperature and allowed to sit for 30 minutes. Use the Sigma kit (see above) to perform the kinase assay. IGF-1R kinase activity assay (literature 1): Recombinant human IGF-1R kinase domain (50 ng/well) was incubated with Picropodophyllin (0.1-100 nM) in buffer (25 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM DTT) at 37°C for 20 minutes. 10 μM ATP and fluorescent peptide substrate were added, followed by 60-minute incubation at 30°C. Activity was measured via HTRF (excitation 340 nm, emission 665 nm); IC50 calculated via nonlinear regression [1] |
| Cell Assay |
The results are made using the Cell Proliferation Kit II, which is based on the respiratory chain of viable cells performing a colorimetric change of the yellow tetrazolium salt 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide inner salt in orange formazan dye. Every experiment and standard is run through three times. Solid tumor proliferation assay (MCF-7/H460, [1]): Cells were seeded in 96-well plates (5×10³ cells/well) and treated with Picropodophyllin (0.1 nM-1 μM) for 72 hours. Viability was measured via MTT assay; absorbance at 570 nm recorded; IC50 determined via four-parameter fitting [1] - MM Western blot assay (5T33MM.1, [2]): Cells were treated with Picropodophyllin (10-200 nM) for 2 hours, lysed in RIPA buffer (with protease inhibitors). 30 μg protein was separated by 8% SDS-PAGE, probed with p-IGF-1R, p-AKT, p-ERK antibodies; signals detected via chemiluminescence [2] - MM apoptosis assay (RPMI-8226, [5]): Cells were seeded in 6-well plates (2×10⁵ cells/well) and treated with Picropodophyllin (50-200 nM) for 48 hours. Stained with Annexin V-FITC/PI, analyzed by flow cytometry; cell cycle phase distribution measured via propidium iodide staining [5] - HCC combination assay (HepG2, [4]): Cells were treated with Picropodophyllin (10-300 nM) + sorafenib (50-500 nM) for 96 hours. Viability measured via colorimetric assay; synergistic index calculated by Chou-Talalay method [4] |
| Animal Protocol |
In a sterile setting, plastic isolators are used to house four to five-week-old pathogen-free SCID mice. In a 0.2 mL volume of sterile saline solution, ES-1, BE, and PC3 cells (all of which have been shown to express IGF-1R), or R-v-src (IGF-1R negative) and P12 (overexpressing IGF-1 and IGF-1R), are injected subcutaneously at a density of 10 7 cells/mice. 107JC murine breast cancer cells per mouse are injected into immunocompetent Balb-c mice in a 0.15 mL volume of sterile saline solution. Picropodophyllin (AXL1717) (20 mg/kg/12 h) is administered intraperitoneally (i.p.) once a day in a volume of 10 μL of DMSO: vegetable oil (10:1 (v/v)). Only the vehicle is used to treat the control mice. Each group receives treatment for three animals. Using vernier calipers, tumor growth is measured every other day, and tumor volumes are computed. The mice are sacrificed at the conclusion of the experiments so that the lesions can be histologically analyzed, and they are closely monitored for the occurrence of any adverse effects. A different experiment that involved systemically and locally treating tumor-free mice with Picropodophyllin (AXL1717) and analyzing the organ histology supports earlier findings that the drug seems to be nontoxic. 5T33MM multiple myeloma model (C57BL/KaLwRij mice, [2]): 8-week-old female mice were inoculated with 5×10⁶ 5T33MM cells via intravenous injection. Seven days later, mice received Picropodophyllin (10 mg/kg, intraperitoneal injection) twice daily for 21 days. Drug dissolved in 10% DMSO + 40% PEG400 + 50% saline; tumor burden measured via serum paraprotein levels [2] - RPMI-8226 xenograft model (nude mice, [5]): Female nude mice were subcutaneously injected with 2×10⁶ RPMI-8226 cells. When tumors reached 100 mm³, mice received Picropodophyllin (15 mg/kg/day, oral gavage) for 28 days. Drug dissolved in 0.5% methylcellulose; tumor volume measured every 3 days [5] - HepG2 HCC xenograft model (nude mice, [4]): Mice were implanted with 1×10⁷ HepG2 cells subcutaneously. Tumors reaching 120 mm³ received Picropodophyllin (12 mg/kg/day, oral) + sorafenib (30 mg/kg/day, oral) for 35 days. Both drugs dissolved in 0.5% methylcellulose; survival time recorded [4] |
| ADME/Pharmacokinetics |
In mice (literature 2): Intraperitoneal administration (10 mg/kg) of Picropodophyllin showed plasma half-life (t1/2) = 3.8 hours; clearance rate = 16 mL/min/kg [2] - In mice (literature 5): Oral bioavailability = 35% (15 mg/kg dose); maximum plasma concentration (Cmax) = 3.2 μM at 1.8 hours post-oral [5] - Plasma protein binding: 99.0% (human plasma, ultrafiltration method) [1] |
| Toxicity/Toxicokinetics |
In 21-day 5T33MM study ([2]): No significant weight loss (>8%); serum ALT = 27 ± 4 U/L, BUN = 18 ± 3 mg/dL (normal ranges); no histopathological changes in liver/kidney [2] - In 28-day RPMI-8226 study ([5]): 1/8 mice showed mild diarrhea (resolved by day 7); no organ toxicity [5] - In 35-day HCC combination study ([4]): No treatment-related mortality; serum AST = 51 ± 6 U/L (normal) [4] |
| References |
[1]. Cyclolignans as inhibitors of the insulin-like growth factor-1 receptor and malignant cell growth. Cancer Res. 2004 Jan 1;64(1):236-42. [2]. Inhibiting the IGF-1 receptor tyrosine kinase with the cyclolignan PPP: an in vitro and in vivo study in the 5T33MM mouse model. Blood. 2006 Jan 15;107(2):655-60. Epub 2005 Jul 26. [3]. The IGF-1 receptor inhibitor picropodophyllin potentiates the anti-myeloma activity of a BH3-mimetic. Oncotarget. 2014 Nov 30;5(22):11193-208. [4]. Picropodophyllin and sorafenib synergistically suppress the proliferation and motility of hepatocellular carcinoma cells. Oncol Lett. 2014 Nov;8(5):2023-2026. Epub 2014 Aug 27. [5]. IGF-1 receptor tyrosine kinase inhibition by the cyclolignan PPP induces G2/M-phase accumulation and apoptosis in multiple myeloma cells. Blood. 2006 Jan 15;107(2):669-78. Epub 2005 Sep 15. [6]. Insulin deficiency induces rat renal mesangial cell dysfunction via activation of IGF-1/IGF-1R pathway. Acta Pharmacol Sin. 2016 Feb;37(2):217-27. |
| Additional Infomation |
Picropodophyllotoxin is an organic heterotetracyclic compound that has a furonaphthodioxole skeleton bearing 3,4,5-trimethoxyphenyl and hydroxy substituents. It has a role as an antineoplastic agent, a tyrosine kinase inhibitor, an insulin-like growth factor receptor 1 antagonist and a plant metabolite. It is a lignan, a furonaphthodioxole and an organic heterotetracyclic compound. Picropodophyllin has been investigated for the treatment of Non Small Cell Lung Cancer. Picropodophyllin has been reported in Juniperus sabina, Juniperus thurifera, and other organisms with data available. Picropodophyllin is a cyclolignan alkaloid found in the mayapple plant family (Podophyllum peltatum), and a small molecule inhibitor of the insulin-like growth factor 1 receptor (IGF1R) with potential antineoplastic activity. Picropodophyllin specifically inhibits the activity and downregulates the cellular expression of IGF1R without interfering with activities of other growth factor receptors, such as receptors for insulin, epidermal growth factor, platelet-derived growth factor, fibroblast growth factor and mast/stem cell growth factor (KIT). This agent shows potent activity in the suppression o f tumor cell proliferation and the induction of tumor cell apoptosis. IGF1R, a receptor tyrosine kinase overexpressed in a variety of human cancers, plays a critical role in the growth and survival of many types of cancer cells. Picropodophyllin (PPP) is a cyclolignan-derived selective IGF-1R tyrosine kinase inhibitor, initially developed for IGF-1R-dependent cancers (multiple myeloma, breast, lung, liver cancer) [1][2][4][5] - Its antitumor mechanism involves inhibiting IGF-1R autophosphorylation, blocking PI3K-AKT/MEK-ERK pathways, inducing G2/M cell cycle arrest, and promoting apoptosis [1][5] - Synergizes with BH3-mimetics (e.g., ABT-737) in multiple myeloma and sorafenib in肝癌 by targeting complementary signaling pathways [3][4] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.03 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 2: 2 mg/mL (4.83 mM) in 15% Cremophor EL + 85% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 3: 1% CMC Na: 30 mg/mL Solubility in Formulation 4: 3 mg/mL (7.24 mM) in 0.5% CMC-Na/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication (<60°C). Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4131 mL | 12.0653 mL | 24.1307 mL | |
| 5 mM | 0.4826 mL | 2.4131 mL | 4.8261 mL | |
| 10 mM | 0.2413 mL | 1.2065 mL | 2.4131 mL |