Genz-644282, formerly known as SAR402674, is a novel, potent and non-camptothecin inhibitor of topoisomerase I with potential antineoplastic activity.The enzyme topoisomerase I is bound to and inhibited by the topoisomerase I inhibitor Genz-644282. This may lead to the inhibition of DNA replication, repair of single-strand breaks in DNA, and growth of tumor cells in tumor cell populations that are susceptible.
Physicochemical Properties
| Molecular Formula | C24H22F3N3O7 |
| Molecular Weight | 521.44 |
| Exact Mass | 407.148 |
| Elemental Analysis | C, 55.28; H, 4.25; F, 10.93; N, 8.06; O, 21.48 |
| CAS # | 529488-28-6 |
| Related CAS # | 529488-28-6;1238632-19-3 (citrate); |
| PubChem CID | 10294813 |
| Appearance | Yellow solid powder |
| LogP | 3.059 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 30 |
| Complexity | 636 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O1COC2C=C3C(=CC1=2)C1=C(C=N3)C2=CC(=C(C=C2C(N1CCN([H])C)=O)OC)OC |
| InChi Key | BAORCAMWLWRZQG-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C22H21N3O5/c1-23-4-5-25-21-14-8-19-20(30-11-29-19)9-16(14)24-10-15(21)12-6-17(27-2)18(28-3)7-13(12)22(25)26/h6-10,23H,4-5,11H2,1-3H3 |
| Chemical Name | 16,17-dimethoxy-21-[2-(methylamino)ethyl]-5,7-dioxa-11,21-diazapentacyclo[11.8.0.02,10.04,8.014,19]henicosa-1(13),2,4(8),9,11,14,16,18-octaen-20-one |
| Synonyms | GENZ-644282; GENZ 644282; GENZ644282; SAR 402674; SAR402674; SAR-402674 |
| 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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 | Topo I (IC50 = 1.2 nM) |
| ln Vitro | Genz-644282 is a topoisomerase I inhibitor. With IC50 values ranging from 1.8 nM to 1.8 μM, Genz-644282 exhibits strong effects against 29 human tumor cell lines[1]. With IC50s ranging from 0.2-21.9 nM and an average IC50 value of 1.2 nM, Genz-644282 suppresses the PPTP cell lines[2]. Strong binding of Top1-DNA covalent cleavage complexes can be achieved with Genz-644282. Human colon cancer HCT116 cells and breast cancer MCF7 cells express γH2AX foci in response to genz-644282 (0.1 μM). On human cancer cell lines that are resistant to CPT, Genz-644282 is cytotoxic[3]. |
| ln Vivo | Genz-644282 (1-4 mg/kg) is active when administered intravenously to the mice. In the human HCT-116 colon cancer xenograft, the human HT-29 colon carcinoma xenograft, and mice harboring the NCI-H460 human non-small cell lung carcinoma, genz-644282 (2.7 mg/kg, i.v.) results in a tumor growth delay (TGD) of 34 days. In the human HCT-15 colon carcinoma xenograft, genz-644282 (2 mg/kg, i.v.) causes a TGD of 33 days; in mice with LOX-IMVI melanoma, it is 28 days. In addition, Genz-644282 (1 mg/kg, i.v.) causes mice harboring the DLD-1 human colon carcinoma xenograft to experience TGD for 14 days. Additionally, Genz-644282 (1.7 mg/kg, i.v.) causes a TGD of 33 days in an NCI-H1299 human non-small cell lung carcinoma xenograft and 23 days in mice with 786-O tumors[1]. In 6/6 evaluable solid tumor models, genz644282 produces maintained complete responses (MCR) at the maximum tolerated dose (MTD, 4 mg/kg). In 3 out of 3 tumor models, Genz644282 (2 mg/kg) induces CR or MCR; in 7 out of 17 (41%) models, it causes objective regressions; however, at 1 mg/kg, there are no objective responses[2]. |
| Cell Assay | Genz-644282 is exposed to 29 established human tumor cell lines over a concentration range in two to four separate experiments. Human tumor cell lines that reflect a variety of histologies and potential resistance mechanisms include MIA PaCa-2, AsPC-1, BxPC-3, CFPAC-1, Hs766T and Capan-1 pancreatic cancers; MEL624, C32, Hs695T and SK-MEL-3 melanomas; NCI-H1299, NCI-H292, NCI-H1915 and SW900 non-small cell lung cancers; HCC1395, HCC1937, HCC202, Hs578T, T-47D and ZR-75-1 breast cancers; ACHN, 769-P, A-498, A-704, SW156, Caki-2 and TK-10 renal cancers; and OVCAR-4 and OVCAR-5 ovarian cancers. In 96-well tissue culture plates, cells are plated at a diameter of 4 × 103/well. The medium used is 100 µL RPMI supplemented with 5% FBS and 12 concentrations of Genz-644282, ranging from 0.1 nM to 10 µM. Each concentration is tested in triplicate. Plates are incubated for one night at 37ºC in 5% CO2 humidified air. For 72 hours, plates are incubated with Genz-644282 at 37ºC in humidified air with 5% CO2. The test plates are read using the Cell Titer-Glo Luminescent Cell Viability Assay following the incubation period. Using a Synergy HT plate reader and the related Ineticalc software, Version #3.4, luminosity is measured. Through comparison with the luminescence of each cell line's untreated control and the determination of IC50 and IC90 values from the graphical data, luminescence data are translated into growth fraction. Two independent experiments are conducted for every cell line[1]. |
| Animal Protocol | Treatments are started when tumors grow to 200 mm3. Nu/nu mice are implanted subcutaneously with a 4 mm3 tumor fragment. Pre-injection preparation involves preparing compounds freshly. For example, Genz-644282 is prepared in M/6 lactate, irinotecan in D5W (5% Dextrose, aqueous), gemcitabine in saline, and docetaxel in ethanol, Cremophor EL, and saline. In experiments using human colon carcinoma xenografts (HCT-116, HT-29, HCT-15, and DLD-1), as well as renal cell carcinoma xenografts (786-O), genz-644282 is compared with irinotecan. Three IV injections of irinotecan at a dose of 60 mg/kg/day are given every four days. A comparison between Genz-644282 and docetaxel is made using the human CIH460 non-small cell lung carcinoma xenograft. Docetaxel is injected intravenously (IV) three times a day at a dose of 12, 16, or 20 mg/kg. A comparison is made between dacarbazine and Genz-644282 in the human LOX-IMVI melanoma xenograft. Dacarbazine is injected intraperitoneally (IP) once daily for five days at a dose of 90 mg/kg/day. In all in vivo experiments, genz-644282 is given intravenously (IV) on alternate days at 1, 1.36, 1.7, 2.7, or 4.1 mg/kg/day for a duration of two weeks[1]. |
| References |
[1]. Genz-644282, a novel non-camptothecin topoisomerase I inhibitor for cancer treatment. Clin Cancer Res. 2011 May 1;17(9):2777-87. [2]. Testing of the topoisomerase 1 inhibitor Genz-644282 by the pediatric preclinical testing program. Pediatr Blood Cancer. 2012 Feb;58(2):200-9 [3]. Molecular and cellular pharmacology of the novel noncamptothecin topoisomerase I inhibitor Genz-644282. Mol Cancer Ther. 2011 Aug;10(8):1490-9. |
| Additional Infomation | Topoisomerase I Inhibitor Genz-644282 is a non-camptothecin inhibitor of topoisomerase I with potential antineoplastic activity. Topoisomerase I inhibitor Genz-644282 binds to and inhibits the enzyme topoisomerase I, which may result in the inhibition of repair of single-strand DNA breaks, DNA replication, and tumor cell growth in susceptible tumor cell populations. |
Solubility Data
| Solubility (In Vitro) | DMSO: ~3.6 mg/mL (~8.9 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 | 1.9178 mL | 9.5888 mL | 19.1777 mL | |
| 5 mM | 0.3836 mL | 1.9178 mL | 3.8355 mL | |
| 10 mM | 0.1918 mL | 0.9589 mL | 1.9178 mL |