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IM156 (IM-156; IM 156; HL-156A; HL156A) is a metformin/biguanide analog with potential anticancer and neuroprotective activity. It works as an OxPhos inhibitor and AMPK activator, increasing AMPK phosphorylation. In an animal model, IM156 lessens cognitive impairment brought on by aging.
Physicochemical Properties
| Molecular Formula | C13H16F3N5O |
| Molecular Weight | 315.3002 |
| Exact Mass | 315.13 |
| Elemental Analysis | C, 49.52; H, 5.12; F, 18.08; N, 22.21; O, 5.07 |
| CAS # | 1422365-93-2 |
| Related CAS # | Lixumistat acetate;1422365-94-3;Lixumistat hydrochloride;1422365-52-3 |
| PubChem CID | 71512108 |
| Appearance | Solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 337.2±52.0 °C at 760 mmHg |
| Flash Point | 157.8±30.7 °C |
| Vapour Pressure | 0.0±0.7 mmHg at 25°C |
| Index of Refraction | 1.578 |
| LogP | 0.96 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 22 |
| Complexity | 424 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | FC(OC1C=CC(=CC=1)/N=C(\N)/N=C(\N)/N1CCCC1)(F)F |
| InChi Key | NGFUHJWVBKTNOE-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C13H16F3N5O/c14-13(15,16)22-10-5-3-9(4-6-10)19-11(17)20-12(18)21-7-1-2-8-21/h3-6H,1-2,7-8H2,(H4,17,18,19,20) |
| Chemical Name | N'-[N'-[4-(trifluoromethoxy)phenyl]carbamimidoyl]pyrrolidine-1-carboximidamide |
| Synonyms | lixumistat; IM156; IM 156; IM-156 |
| 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 | AMPK, OXPHOS |
| ln Vitro |
IM156 (0.31-10 μM) phosphorylates AMPK1 Thr172 in NIH3T3 mouse fibroblast cells in a dose- and time-dependent manner[1]. Glucose-6-phosphatase (G6pase) and phosphoenolpyruvate carboxykinase 1 (Pck1), two important players in glucose homeostasis, are not affected by IM156 on their expression[1]. |
| ln Vivo |
IM156 has no effect on metabolic control as measured by body weight, blood sugar, insulin levels, and lipid metabolite content in mice with diet-induced obesity[1].
Anxiety, general locomotion, or body weight are unaffected by IM156 (50 mg/kg; for 2 months)[2]. IM156 (50 mg/kg; for 2 months) does not affect body weight, general locomotion, or anxiety[2]. IM156 significantly slows the aging-related decline in spatial working memory and novel object recognition memory[2]. In the hippocampus of old mice, IM156 significantly boosts AMPK activation[2]. |
| Enzyme Assay | A six-well plate is seeded with RPMCs at a density of 1 105 cells per well. RPMCs are grown to 70% confluence and serum-starved for 24 hours while under NG conditions. Using a siRNA transfection reagent and following the manufacturer's instructions, the specific AMPKα1 small interfering (si)RNA, AMPKα2 siRNA, or control siRNA are transfected into the cells. Using siRNA dilution buffer, AMPKα1 small interfering (si)RNA, AMPKα2 siRNA are incubated for 30 min at room temperature with a final concentration of 20 M. 2 ml of siRNA transfection medium is used to wash the cells twice, and then 200 μl of siRNA transfection reagent complex and 0.8 ml of siRNA transfection medium are added to the well, covering the entire layer. To ensure that the entire cell layer is submerged in the solution, the contents of the plate are gently swirled. The cells are then placed in an incubator with 5% CO2 for 5 hours at 37°C. One milliliter of regular growth medium is then added. The cells are then incubated for a further 24 hours. The media are then swapped out for brand-new, finished ones and treated with NG or HG for 24 hours in either IM156 (HL156A) presence or absence. |
| Animal Protocol |
5 to 6-wk-old female C57BL/6 mice 5 mg/kg IP |
| References |
[1]. HL271, a novel chemical compound derived from metformin, differs from metformin in its effects on the circadian clock and metabolism. Biochem Biophys Res Commun. 2016 Jan 15;469(3):783-9. [2]. The Improving Effect of HL271, a Chemical Derivative of Metformin, a Popular Drug for Type II Diabetes Mellitus, on Aging-induced Cognitive Decline. Exp Neurobiol. 2018 Feb;27(1):45-56. [3]. Phase I study of IM156, a novel potent biguanide oxidative phosphorylation (OXPHOS) inhibitor, in patients with advanced solid tumors. Journal of Clinical Oncology 38(15_suppl):3590-3590. |
| Additional Infomation | Lixumistat is an orally bioavailable biguanide compound and mitochondrial oxidative phosphorylation (OxPhos) inhibitor, with potential antineoplastic activity. Upon administration, lixumistat inhibits oxidative phosphorylation, decreases mitochondrial function, prevents tumor cell metabolism and deprives tumor cells of energy, thereby preventing tumor cell proliferation. Mitochondrial OxPhos is overactivated in cancer cells and plays a key role in tumor cell proliferation. Drug resistant tumor cells are very susceptible to decreased mitochondrial OxPhos as they cannot easily compensate for the decrease in mitochondrial function by increasing glycolysis. |
Solubility Data
| Solubility (In Vitro) | DMSO: 63~100 mg/mL (199.8~266.4 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 | 3.1716 mL | 15.8579 mL | 31.7158 mL | |
| 5 mM | 0.6343 mL | 3.1716 mL | 6.3432 mL | |
| 10 mM | 0.3172 mL | 1.5858 mL | 3.1716 mL |