Ombrabulin HCl (AV-E8062; AC-7700; AVE8062), an analogue of combretastatin A4 with improved water solubility, is a novel and potent tubulin inhibitor and vascular-disrupting agent with anticancer activity.It has been the subject of eleven completed phase studies and has better solubility, oral bioavailability, improved anti-cancer activity, and decreased toxicity.
Physicochemical Properties
| Molecular Formula | C21H27CLN2O6 |
| Molecular Weight | 438.90188 |
| Exact Mass | 438.156 |
| Elemental Analysis | C, 57.47; H, 6.20; Cl, 8.08; N, 6.38; O, 21.87 |
| CAS # | 253426-24-3 |
| Related CAS # | Ombrabulin;181816-48-8 |
| PubChem CID | 6918404 |
| Appearance | White to off-white solid powder |
| LogP | 4.301 |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 9 |
| Heavy Atom Count | 30 |
| Complexity | 517 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | Cl[H].O(C([H])([H])[H])C1C([H])=C([H])C(/C(/[H])=C(/[H])\C2C([H])=C(C(=C(C=2[H])OC([H])([H])[H])OC([H])([H])[H])OC([H])([H])[H])=C([H])C=1N([H])C([C@]([H])(C([H])([H])O[H])N([H])[H])=O |
| InChi Key | UQNRTPFLTRZEIM-MRWUDIQNSA-N |
| InChi Code | InChI=1S/C21H26N2O6.ClH/c1-26-17-8-7-13(9-16(17)23-21(25)15(22)12-24)5-6-14-10-18(27-2)20(29-4)19(11-14)28-3;/h5-11,15,24H,12,22H2,1-4H3,(H,23,25);1H/b6-5-;/t15-;/m0./s1 |
| Chemical Name | (2S)-2-amino-3-hydroxy-N-[2-methoxy-5-[(Z)-2-(3,4,5-trimethoxyphenyl)ethenyl]phenyl]propanamide;hydrochloride |
| Synonyms | AVE8062; AC 7700; CS-39-L-Ser.HCl; AVE8062; AC7700; AVE8062; AVE-8062A; Ombrabulin; AC-7700; Ombrabulin HCl |
| 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
| Targets | tubulin |
| ln Vitro | The MTT assay is utilized to investigate the impact of Ombrabulin (AVE8062) on the viability of tumor or endothelial cells. For the tumor cell lines (HeyA8, SKOV3ip1, and HeyA8-MDR), ombrabulin's half-life (IC50) is 7–20 nM, while it is 10 nM for mouse mesenteric endothelial cells (MMEC). Ombrabulin / Docetaxel combination dose-response curves show a significantly lower IC50 than either agent alone (P<0.005, all cell lines) when compared to each agent alone in the nonlinear least-squares regression analysis. Compared to docetaxel alone, the cytotoxicity of docetaxel to tumor and endothelial cells is two to four times higher when combined with ombrabium[1]. |
| ln Vivo | In nude mice (n=3 per group), the tolerability of different doses of Ombrabulin (AVE8062) ranging from 10 to 100 mg/kg is tested twice weekly via i.v., i.p., or s.c. routes prior to conducting therapy experiments. Due to issues with skin or tail vein necrosis, the i.v. and s.c. routes are not pursued further. With dosages up to 100 mg/kg, the intraperitoneal route is well tolerated. The lowest dose for in vivo therapeutic efficacy is then determined by preliminary studies. Three weeks after the injection of tumor cells, nude mice (n = 5 per group) injected with HeyA8 ovarian cancer cells are given vehicle or Ombrabulin 10, 30, 50, and 100 mg/kg intraperitoneally twice a week for three weeks. In comparison to the vehicle control group, the tumor weight is reduced by 65% in the 30 mg/kg group (P<0.02). No effect is seen with a dose of 10 mg/kg. For further therapy trials, the 30 mg/kg dose is chosen because the antitumor effects at doses higher than 30 mg/kg are not appreciably better[1]. |
| Cell Assay | Flow cytometry is used to assess Ombrabulin (AVE8062)'sabilityto modify the HeyA8 and MMEC cell cycles as well as apoptosis. Petri dishes containing 3×106 tumor cells are seeded and left to adhere overnight for each assay. Next, the cultures are treated with regular medium (negative control), medium containing Docetaxel, medium containing Ombrabulin (HeyA8, 20 nM; MMEC 10 nM), or medium containing Ombrabulin plus Docetaxel. The cultures are then rinsed with PBS. Trypsinization is used to gather tumor cells, which are then combined with cells floating in the medium for cell cycle analysis. After washing and fixing with ethanol, the cell suspensions are centrifuged for five minutes at 1,500 rpm at room temperature. In order to analyze apoptosis, cells are incubated at room temperature for one night in 50 μL of DNA labeling solution, which consists of 10 μL of reaction buffer, 0.75 μL of TdT enzyme, 8 μL of FITC-dUTP, and 32.25 μL of distilled water. After adding rinse buffer, samples are cleaned, fixed in ethanol, and centrifuged. Following a PBS wash, all samples are resuspended in PBS containing 50 μg/mL of propidium iodide and 20 μg/mL of RNase A, and left to sit at room temperature for 30 minutes. A flow cytometer called an EPICS XL is used to analyze stained cells. The low-level gate is positioned at the base of the G1 peak, and Multicycle analysis is used to ascertain the percentages of cells in the G1 and G2-M phases of the cell cycle[1]. |
| Animal Protocol | Mice: The mice used are 6–8 week old female athymic nude mice. To be injected into a living being, the tumor cells undergo trypsinization, centrifugation at 1,000 rpm for 7 minutes at 4°C, two washings, and resuspension in serum-free HBSS at a density of 1.25×106 cells/mL (HeyA8) and 5×106 cells/mL (SKOV3ip1 and HeyA8-MDR). Cells are injected intraperitoneally (i.p.) to form tumors. After injecting cell lines, ombrabulin therapy is started seven or seventeen days later. Mice (n = 10 per group) are randomized into the following treatment groups: (a) weekly intraperitoneal injection (PBS) of 200 μL; (b) twice weekly intraperitoneal injection (Ombrabulin 30 mg/kg [dissolved in PBS (pH 5)]; (c) weekly intraperitoneal injection (Docetaxel 2 mg/kg (HeyA8 and HeyA8-MDR) or 1.4 mg/kg (SKOV3ip1); and (d) weekly injection of Ombrabulin plus Docetaxel (both drugs given at the above doses and frequencies for each drug alone). The Ombrabulin dosage employed in these investigations was refined through dose-escalation investigations to inhibit the growth of tumors. Tumors are removed from mice after treatment (range 2–5 weeks), and they are watched for any signs of negative side effects. At necropsy, the weight of the tumor, the volume of ascites, the number of tumor nodules, and the mouse weight are all noted. |
| References |
[1]. Antitumor and antivascular effects of AVE8062 in ovarian carcinoma. Cancer Res. 2007 Oct 1;67(19):9337-45. |
Solubility Data
| Solubility (In Vitro) | H2O: ~20 mg/mL (~45.6 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 | 2.2784 mL | 11.3921 mL | 22.7842 mL | |
| 5 mM | 0.4557 mL | 2.2784 mL | 4.5568 mL | |
| 10 mM | 0.2278 mL | 1.1392 mL | 2.2784 mL |