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SNS-032 (also known as BMS-387032; SNS 032; BMS387032; SNS032), a 2-aminothiazole based small-molecule, is a potent and selective inhibitor of CDK2 (cyclin dependent kinase 2) with potential antitumor activity. In cell-free experiments, it inhibits CDK2 with an IC50 of 48 nM and exhibits 10- and 20-fold selectivity over CDK1/CDK4. SNS-032 binds specifically to CDKs 2, 7, and 9, inhibiting their phosphorylation and activation. In tumor cell populations that are susceptible, this inhibition of CDK activity may lead to cell cycle arrest, apoptosis induction, and decreased tumor cell proliferation. There is evidence that this agent can make radioresistant tumor cells more susceptible to ionizing radiation.
Physicochemical Properties
| Molecular Formula | C17H24N4O2S2 |
| Molecular Weight | 380.53 |
| Exact Mass | 380.134 |
| Elemental Analysis | C, 53.66; H, 6.36; N, 14.72; O, 8.41; S, 16.85 |
| CAS # | 345627-80-7 |
| Related CAS # | 345627-80-7;345627-90-9 (HCl); |
| PubChem CID | 3025986 |
| Appearance | white solid powder |
| Density | 1.3±0.1 g/cm3 |
| Index of Refraction | 1.607 |
| LogP | 2.79 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 25 |
| Complexity | 454 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | S1C(=C([H])N=C1N([H])C(C1([H])C([H])([H])C([H])([H])N([H])C([H])([H])C1([H])[H])=O)SC([H])([H])C1=NC([H])=C(C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H])O1 |
| InChi Key | OUSFTKFNBAZUKL-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C17H24N4O2S2/c1-17(2,3)12-8-19-13(23-12)10-24-14-9-20-16(25-14)21-15(22)11-4-6-18-7-5-11/h8-9,11,18H,4-7,10H2,1-3H3,(H,20,21,22) |
| Chemical Name | N-[5-[(5-tert-butyl-1,3-oxazol-2-yl)methylsulfanyl]-1,3-thiazol-2-yl]piperidine-4-carboxamide |
| Synonyms | BMS-387032; SNS 032; BMS387032; SNS032; N-(5-(((5-(tert-Butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-4-carboxamide; N-[5-[(5-tert-butyl-1,3-oxazol-2-yl)methylsulfanyl]-1,3-thiazol-2-yl]piperidine-4-carboxamide; SNS-032 (BMS-387032); BMS 387032; SNS-032 |
| 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 | CDK9 (IC50 = 4 nM); CDK2 (IC50 = 38 nM); CDK7 (IC50 = 62 nM); CDK1 (IC50 = 480 nM); CDK4 (IC50 = 925 nM) |
| ln Vitro | SNS-032 exhibits low sensitivity to CDK1, with an IC50 of 480 nM, and CDK4 at 925 nM. In vitro, SNS-032 efficiently eradicates cells with chronic lymphocytic leukemia, irrespective of treatment history and prognostic markers. SNS-032 is more effective than flavopiridol and roscovitine in both RNA synthesis inhibition and apoptosis induction. The activity of SNS-032 is easily reversible; SNS-032 removal reactivates RNA polymerase II, which results in Mcl-1 resynthesis and cell survival[1]. SNS-032 prevents endothelial cells from forming three-dimensional capillary networks. SNS-032 totally stops HUVECs from forming capillaries through U87MG cell-mediated mechanisms. Furthermore, SNS-032 significantly inhibits both cell lines' ability to produce VEGF, which is linked to its ability to inhibit in vitro angiogenesis. Preclinical research has demonstrated that SNS-032 causes apoptosis and cell cycle arrest in a variety of cell lines[2]. SNS-032 inhibits CDKs 2 and 7 to stop the cell cycle, and CDKs 7 and 9 to inhibit transcription. Human serum has no effect on SNS-032 activity[3]. SNS-032 causes caspase-3 activation and annexin V staining to increase in a dose-dependent manner. SNS-032 inhibits the expression of CDK2 and CDK9 and dephosphorylates CDK7, while at the molecular level it causes a marked dephosphorylation of RNA polymerase (RNA Pol) II's serine 2 and 5[5]. |
| ln Vivo |
SNS-032 (15 mg/kg, i.p.) inhibits KBM5-T315I cells as well as xenografted BaF3-T674I cells in vivo. When T674I PDGFRα and T315I-Bcr-Abl are downregulated in tumors transplanted into nude mice, SNS-032 stops the tumors from growing[4]. SNS-032 improves liver function and fibrotic stage in vivo[6] Male C57BL/6 mice were intraperitoneally injected with 2 ml·kg−1 15% CCl4–olive oil on Mondays, Wednesdays, and Fridays for 6 weeks to establish a murine model of hepatic fibrosis. After 3 weeks of injection, the mice were treated with 2.5, 5, or 10 mg·kg−1 SNS-032 and 5 mg·kg−1 sorafenib for 3 weeks (Figure 1A). Then, the sera and liver tissues of each group were collected. We found that the liver weight and liver/body weight ratio of the mice in the experimental group had significantly increased compared with those of the mice in the normal group (p < 0.001). Liver weight and liver/body weight ratio prominently decreased after treatment with 2.5, 5, or 10 mg·kg−1 SNS-032 and 5 mg·kg−1 sorafenib. After treatment with 5 and 10 mg·kg−1 SNS-032, collagen deposition, inflammatory cell infiltration, and liver fibrosis degree significantly decreased (Figure 1D). The semiquantitatively analyzed Sirius red positive area and hydroxyproline content in liver tissue significantly increased in the experimental group but decreased in the 5 and 10 mg·kg−1 SNS-032 and 5 mg·kg−1 sorafenib groups. The percentage of the Acta2-positive area significantly increased in the experimental group but decreased in the 5 and 10 mg·kg−1 SNS-032 and 5 mg·kg−1 sorafenib groups. In terms of biochemical profiles, the serum levels of ALT and AST were significantly elevated in the experimental group and significantly decreased in the 5 and 10 mg·kg−1 SNS-032 and 5 mg·kg−1 sorafenib groups. |
| Enzyme Assay |
SNS-032 exhibits selectivity against CDK2, CDK7, and CDK9, as evidenced by its IC50 values of 38 nM, 62 nM, and 4 nM, respectively. Cell proliferation, migration, and capillary network formation of endothelial cells are the fundamental steps for angiogenesis, which involves the formation of new blood vessels. The purpose of this study is to investigate the effect of a novel aminothiazole SNS-032 on these critical steps for in vitro angiogenesis using a coculture system consisting of human umbilical vein endothelial cells (HUVECs) and human glioblastoma cells (U87MG). SNS-032 is a potent selective inhibitor of cyclin-dependent kinases 2, 7, and 9, and inhibits both transcription and cell cycle. In this study, we examined the proliferation and viability of HUVECs and U87MG cells in the presence of SNS-032 and observed a dose-dependent inhibition of cellular proliferation in both cell lines. SNS-032 inhibited threedimensional capillary network formations of endothelial cells. In a coculture study, SNS-032 completely prevented U87MG cell-mediated capillary formation of HUVECs. This inhibitor also prevented the migration of HUVECs when cultured alone or cocultured with U87MG cells. In addition, SNS-032 significantly prevented the production of vascular endothelial growth factor (VEGF) in both cell lines, whereas SNS-032 was less effective in preventing capillary network formation and migration of endothelial cells when an active recombinant VEGF was added to the medium. In conclusion, SNS-032 prevents in vitro angiogenesis, and this action is attributable to blocking of VEGF.[2] |
| Cell Assay |
The growth curves of both HUVECs and U87MG cells are measured using the Cell Titer-Glo (CTG) luminescent assay. A total of 100 milliliters is used to seed U87MG cells and HUVECs (2×103 cells/well) in a 96-well microplate. Cells are exposed to different concentrations of SNS-032 (0-0.5 mM) for 24, 48, or 72 hours following the 24-hour mark. Following the treatment's completion, 100 mL of CTG solution is added to each well, and the wells are then left in the dark and at room temperature for 20 minutes. The lysate (50 mL) is put into a 96-well white plate, and POLARstar OPTIMA is used to measure luminescence. When calculating percentage cell growth, 100% growth at the time of SNS-032 addition is taken into account. The effects of SNS-032 on PDGFRα and Bcr-Abl signaling pathways, apoptosis, and cell cycling were analyzed in TKI-resistant cells of HES and CML. The in vivo antitumor activity of SNS-032 was assessed with xenografted BaF3-T674I FIP1L1-PDGFRα and KBM5-T315I Bcr-Abl cells in nude mouse models. Results: SNS-032 inhibited the phosphorylation on Ser5 and Ser2 of RNA polymerase II. SNS-032 decreased both the mRNA and protein levels of FIP1L1-PDGFRα and Bcr-Abl and inhibited the proliferation of malignant cells expressing FIP1L1-PDGFRα or Bcr-Abl. It also decreased the phosphorylation of downstream molecules. It induced apoptosis by triggering both the mitochondrial pathway and the death receptor pathway. Conclusions: This CDK7/9 inhibitor potently inhibits FIP1L1-PDGFRα-positive HES cells and Bcr-Abl-positive CML cells regardless of their sensitivity to imatinib. SNS-032 may have potential in treating hematologic malignancy by abrogating oncogene addiction.[4] Proliferation and colony forming assays were used to evaluate cytotoxicity, Western blot analyses to evaluate target modulation, FACS analysis to assess cell cycle distribution, RT-PCR to evaluate transcriptional inhibition. Results: SNS-032 blocks the cell cycle via inhibition of CDKs 2 and 7, and transcription via inhibition of CDKs 7 and 9. Treatment of RPMI-8226 MM cells at 300 nM (IC(90)) for 6 h was sufficient for commitment to apoptosis. This correlated with inhibition of CDKs 2, 7 and 9, as reflected in substrate signaling molecules. SNS-032 activity was unaffected by human serum. Target modulation was observed in PBMC from treated patients. Conclusions: These results demonstrate SNS-032 target modulation of CDKs 2, 7 and 9, and establish 6 h exposure as sufficient to commit RPMI-8226 MM cells to apoptosis. Combined with the demonstration of target modulation in PBMC from phase 1 solid tumor patients treated with SNS-032, these data support the ongoing clinical study of SNS-032 in MM and CLL.[3] |
| Animal Protocol |
Simple nu/nu In barrier facilities with a 12-hour light-dark cycle, BALB/c mice are kept with unlimited access to food and water. On the flanks of 4- to 6-week-old male nude mice, a mixture of 1×107 BaF3-T674I cells with Matrigel or KBM5-T315I cells (3×107) is subcutaneously injected. Calipers are used every other day to measure tumors. The formula for calculating tumor volumes is a2×b×0.4, where a is the diameter that is the smallest and b is the diameter that is perpendicular to a. After the subcutaneous inoculation, mice are randomized to receive treatment with either vehicle (tissue culture medium containing 0.1% v/v) or SNS-032 (15 mg/kg injected intraperitoneally every two days) for approximately two weeks, or until the tumors are palpable (approximately 100 mm3). Prior to dilution, SNS-032 is dissolved in tissue culture grade DMSO. Each animal's body weight, eating habits, and level of motor activity are tracked as measures of overall health. Tumor xenografts are then promptly removed, weighed, stored, and fixed after the animals are put to sleep. Forty-eight male C57BL6 mice (6-week-old, 18–20 g) were caged individually in a temperature- and humidity-controlled environment under a 12:12 light–dark cycle. Then, they were assigned into two groups randomly. The first group (control group, n = 8) was fed with normal diet and water freely. The second group (n = 40) was intraperitoneally injected with 2 ml·kg−1 15% carbon tetrachloride (CCl4)–olive oil on Mondays, Wednesdays, and Fridays for 3 weeks. Then, the mice in the second group were randomly allocated into five groups (experimental, low-dose SNS-032, medium-dose SNS-032, high-dose SNS-032, and sorafenib groups; n = 8). Subsequently, the mice in the low-, medium-, and high-dose SNS-032 and sorafenib groups were treated with 2.5, 5, and 10 mg·kg−1·day SNS-032 through intraperitoneal administration or 5 mg·kg−1·day sorafenib through intragastric administration for 3 weeks with the continuous injection of CCl4–olive oil. The CCl4 mouse model was used as the liver fibrosis model as previously described (Strnad et al., 2008).[6] |
| References |
[1]. Mechanism of action of SNS-032, a novel cyclin-dependent kinase inhibitor, in chronic lymphocytic leukemia. Blood. 2009 May 7;113(19):4637-45. [2]. SNS-032 prevents tumor cell-induced angiogenesis by inhibiting vascular endothelial growth factor. Neoplasia. 2007 May;9(5):370-81. [3]. SNS-032 is a potent and selective CDK 2, 7 and 9 inhibitor that drives target modulation in patient samples. Cancer Chemother Pharmacol. 2009 Sep;64(4):723-32. [4]. Cyclin-dependent kinase 7/9 inhibitor SNS-032 abrogates FIP1-like-1 platelet-derived growth factor receptor α and bcr-abl oncogene addiction in malignant hematologic cells.Clin Cancer Res. 2012 Apr 1;18(7):1966-78. [5]. The cyclin-dependent kinase inhibitor SNS-032 has single agent activity in AML cells. Leukemia. 2011 Mar;25(3):411-9. [6]. SNS-032 attenuates liver fibrosis by anti-active hepatic stellate cells via inhibition of cyclin dependent kinase 9. Front Pharmacol . 2022 Oct 12:13:1016552. |
| Additional Infomation |
N-(5-{[(5-tert-butyl-1,3-oxazol-2-yl)methyl]sulfanyl}-1,3-thiazol-2-yl)piperidine-4-carboxamide is a secondary carboxamide resulting from the formal condensation of the carboxy group of piperidine-4-carboxylic acid with the amino group of 5-{[(5-tert-butyl-1,3-oxazol-2-yl)methyl]sulfanyl}-1,3-thiazol-2-amine. It is an ATP-competitive inhibitor of CDK2, CDK7 and CDK9 kinases and exhibits anti-cancer properties. It has a role as an apoptosis inducer, an antineoplastic agent, an EC 2.7.11.22 (cyclin-dependent kinase) inhibitor and an angiogenesis inhibitor. It is a piperidinecarboxamide, a member of 1,3-oxazoles, a member of 1,3-thiazoles, an organic sulfide and a secondary carboxamide. CDK Inhibitor SNS-032 is a small aminothiazole molecule and cyclin dependent kinase (CDK) inhibitor with potential antineoplastic activity. SNS-032 binds to and prevents the phosphorylation of cyclin-dependent kinases, especially CDK2, 7, and 9 that regulate cell cycle progression. Inhibition of CDKs leads to cell cycle arrest and induces apoptosis. As a result, this agent causes cytotoxicity and prevents further tumor cell growth. Drug Indication Investigated for use/treatment in cancer/tumors (unspecified), leukemia (unspecified), lymphoma (unspecified), multiple myeloma, and solid tumors. Mechanism of Action SNS-032 is a potent and selective inhibitor of CDKs 2, 7 and 9, which are critical in the communication and relay of signals to promote cellular growth and function. CDK2 is involved in cellular proliferation by regulating the initiation of and progression through the DNA-synthesis phase of the cell cycle. CDK7 and CDK9 are involved in transcriptional regulation of certain proteins involved in cell survival. Inappropriate activity by these CDKs can lead to unregulated proliferation, avoidance of apoptosis and increased cell survival, all of which are hallmarks of cancer. By selectively targeting these CDKs, SNS-032 may halt both aberrant cell proliferation and induce apoptosis. Inhibitors of cyclin-dependent kinases (Cdks) have been reported to have activities in chronic lymphocytic leukemia cells by inhibiting Cdk7 and Cdk9, which control transcription. Here we studied the novel Cdk inhibitor SNS-032, which exhibits potent and selective inhibitory activity against Cdk2, Cdk7, and Cdk9. We hypothesized that transient inhibition of transcription by SNS-032 would decrease antiapoptotic proteins, resulting in cell death. SNS-032 effectively killed chronic lymphocytic leukemia cells in vitro regardless of prognostic indicators and treatment history. This was associated with inhibition of phosphorylation of RNA polymerase II and inhibition of RNA synthesis. Consistent with the intrinsic turnover rates of their transcripts and proteins, antiapoptotic proteins, such as Mcl-1 and X-linked inhibitor of apoptosis protein (XIAP), were rapidly reduced on exposure to SNS-032, whereas Bcl-2 protein was not affected. The initial decrease of Mcl-1 protein was the result of transcriptional inhibition rather than cleavage by caspase. Compared with flavopiridol and roscovitine, SNS-032 was more potent, both in inhibition of RNA synthesis and at induction of apoptosis. SNS-032 activity was readily reversible; removal of SNS-032 reactivated RNA polymerase II, which led to resynthesis of Mcl-1 and cell survival. Thus, these data support the clinical development of SNS-032 in diseases that require short-lived oncoproteins for survival.[1] SNS-032 (BMS-387032) is a selective cyclin-dependent kinase (CDK) inhibitor. In this study, we evaluated its effects on primary acute myeloid leukemia (AML) samples (n=87). In vitro exposure to SNS-032 for 48 h resulted in a mean LD(50) of 139±203 nM; Cytarabine (Ara-C) was more than 35 times less potent in the same cohort. SNS-032-induced a dose-dependent increase in annexin V staining and caspase-3 activation. At the molecular level, SNS-032 induced a marked dephosphorylation of serine 2 and 5 of RNA polymerase (RNA Pol) II and inhibited the expression of CDK2 and CDK9 and dephosphorylated CDK7. Furthermore, the combination of SNS-032 and Ara-C showed remarkable synergy that was associated with reduced mRNA levels of the antiapoptotic genes XIAP, BCL2 and MCL1. In conclusion, SNS-032 is effective as a single agent and in combination with Ara-C in primary AML blasts. Treatment with Ara-C alone significantly induced the transcription of the antiapoptotic genes BCL2 and XIAP. In contrast, the combination of SNS-032 and Ara-C suppressed the transcription of BCL2, XIAP and MCL1. Therefore, the combination of SNS-032 and Ara-C may increase the sensitivity of AML cells to the cytotoxic effects of Ara-C by inhibiting the transcription of antiapoptotic genes.[5] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.47 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (5.47 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (5.47 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 30% PEG400+0.5% Tween80+5% Propylene glycol : 30 mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6279 mL | 13.1396 mL | 26.2791 mL | |
| 5 mM | 0.5256 mL | 2.6279 mL | 5.2558 mL | |
| 10 mM | 0.2628 mL | 1.3140 mL | 2.6279 mL |