TAME (Tosyl-L-Arginine Methyl Ester) is a potent small molecule APC (anaphase-promoting complex/cyclosome) inhibitor, which can target proteins for degradation and cause chromatid separation.
Physicochemical Properties
| Molecular Formula | C14H22N4O4S | |
| Molecular Weight | 342.41 | |
| Exact Mass | 342.136 | |
| CAS # | 901-47-3 | |
| Related CAS # | 1784-03-8 | |
| PubChem CID | 1550286 | |
| Appearance | White to off-white solid | |
| Density | 1.3±0.1 g/cm3 | |
| Boiling Point | 516.5±60.0 °C at 760 mmHg | |
| Flash Point | 266.1±32.9 °C | |
| Vapour Pressure | 0.0±1.3 mmHg at 25°C | |
| Index of Refraction | 1.591 | |
| LogP | 0.77 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 6 | |
| Rotatable Bond Count | 9 | |
| Heavy Atom Count | 23 | |
| Complexity | 503 | |
| Defined Atom Stereocenter Count | 1 | |
| SMILES | S(C1C=CC(C)=CC=1)(=O)(=O)N[C@H](C(=O)OC)CCCNC(N)=N |
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| InChi Key | FKMJXALNHKIDOD-LBPRGKRZSA-N | |
| InChi Code | InChI=1S/C14H22N4O4S/c1-10-5-7-11(8-6-10)23(20,21)18-12(13(19)22-2)4-3-9-17-14(15)16/h5-8,12,18H,3-4,9H2,1-2H3,(H4,15,16,17)/t12-/m0/s1 | |
| Chemical Name | methyl (2S)-5-(diaminomethylideneamino)-2-[(4-methylphenyl)sulfonylamino]pentanoate | |
| Synonyms |
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| 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 |
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| 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 | Anaphase-promoting complex (APC) | ||
| ln Vitro | TAME has an IC50 of 12 µM for inhibiting cyclin proteolysis in mitotic Xenopus egg extract. Interphase extract treated with recombinant cyclin B1/Cdc2 complex in mitosis is stopped by TAME at concentrations of 1-200 μM, leaving phosphorylated Cdc27 and stable cyclin B1. When TAME is added at a 200 μM concentration, it significantly reduces Cdh1's binding to the Anaphase-Promoting Complex (APC) and inhibits its ubiquitin ligase activity. The C-terminal isoleucine-arginine (IR) tail of APC is the contribution motif that TAME addition to interphase extract uses to reduce Cdc20 association with the APC in a dose-dependent manner. Trypsin hydrolyzes TAME, with a Km of 0.328 mM.[2] TAME causes an approximate 12-fold increase in ATP hydrolysis speed.[3] TAME interacts with ATP's β and γ phosphate as well as the adenine ring through the aromatic ring and guanidinium group.[4] TAME inhibits Bacillus subtilis nutrient-induced germination and pressure-induced germination at 600 MPa at a concentration of 50 mM.[5] On ileal strips, TAME causes a concentration-dependent contractile response with an EC50 of 4.3 x 103 M.[6] | ||
| ln Vivo |
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| Enzyme Assay | TAME at concentrations ranging from 1 to 200 μM stops interphase extract treated with recombinant cyclin B1/Cdc2 complex in mitosis, resulting in phosphorylated Cdc27 and stable cyclin B1. At a 200 μM concentration, TAME significantly suppresses the ubiquitin ligase activity of the Anaphase-Promoting Complex (APC), which is accompanied by a decrease in Cdh1 binding to APC. By binding directly to APC, TAME addition to interphase extract reduces Cdc20 association with the APC in a dose-dependent manner. The C-terminal isoleucine-arginine (IR) tail on APC is the contribution motif. Trypsin hydrolyzes TAME, and its Km value is 0.328 mM. TAME speeds up the hydrolysis of ATP by a factor of twelve. Through the guanidinium group and the aromatic ring, TAME interacts with β and γ phosphate as well as the adenine ring of ATP. TAME inhibits both pressure-induced germination at 600 MPa and nutrient-induced germination in Bacillus subtilis at concentrations of 50 mM. On ileal strips, TAME causes a concentration-dependent contractile response with an EC50 of 4.3 x 103 M. | ||
| Cell Assay | In order to prevent APC-dependent proteolysis in mitotic Xenopus oocytes, TAME competes with the Cdc20 C-terminal IR-tail for APC binding. Additionally, TAME stabilizes cyclin B1 by prematurely stopping ubiquitination. It inhibits the first round of ubiquitination of unmodified cyclin B1. Unubiquitinated cyclin B1 is unable to encourage Cdc20 binding to the APC when TAME is present. | ||
| Animal Protocol |
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| References |
[1]. Pharmacologic inhibition of the anaphase-promoting complex induces a spindle checkpoint-dependent mitotic arrest in the absence of spindle damage. Cancer Cell. 2010 Oct 19;18(4):382-95. [2]. An APC/C inhibitor stabilizes cyclin B1 by prematurely terminating ubiquitination. Nat Chem Biol. 2012 Feb 26;8(4):383-92. |
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| Additional Infomation |
TAMe is an L-arginine ester that is methyl L-argininate in which one of the hydrogens attached to the alpha-nitrogen is substituted by a tosyl group. It is a L-arginine ester, a sulfonamide, a methyl ester and a member of guanidines. Arginine derivative which is a substrate for many proteolytic enzymes. As a substrate for the esterase from the first component of complement, it inhibits the action of C(l) on C(4). |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.30 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 25.0 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: ≥ 2.5 mg/mL (7.30 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 25.0 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: ≥ 2.5 mg/mL (7.30 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9205 mL | 14.6024 mL | 29.2048 mL | |
| 5 mM | 0.5841 mL | 2.9205 mL | 5.8410 mL | |
| 10 mM | 0.2920 mL | 1.4602 mL | 2.9205 mL |