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| Targets |
MRT-199665 targets MARK (MARK1/MARK2/MARK3/MARK14), SIK (SIK1/SIK2/SIK3), and AMPK (AMPKα1/AMPKα2) kinases. It is a potent and ATP-competitive inhibitor with IC₅₀ values of 2/2/3/2 nM for MARK1/MARK2/MARK3/MARK14, 10/10 nM for AMPKα1/AMPKα2, and 110/12/43 nM for SIK1/SIK2/SIK3, respectively. By inhibiting these kinases, MRT-199665 modulates inflammatory responses, including increasing IL-10 production and suppressing pro-inflammatory cytokines. The compound is a valuable tool for studying the role of these kinases in inflammation and immune regulation.
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| ln Vitro |
MRT199665 (1 μM; 1 hour post-death) enhances the synthesis of IL-10 and Nurr77 mRNA in response to LPS (100 ng/mL; stimulation lasting up to 24 hours) [1]. Treatment with MRT199665 inhibits the phosphorylation of MEF2C S222 in acute myeloid leukemia (AML) cells. MRT199665 inhibits the growth of leukemia by causing total MEF2C at 10 nM-1000 nM for 12 hours and -100 μM for 48 hours [2]. Additionally, MRT199665 lowers the amount of MEF2C protein overall [2].
In vitro, MRT-199665 potently inhibits MARK, SIK, and AMPK kinase activity in cell-free assays. In cell-based assays, MRT199665 (1 µM; 1 hour post-death) enhances the synthesis of IL-10 and Nur77 mRNA in response to LPS (100 ng/mL; stimulation lasting up to 24 hours). Treatment with MRT199665 increases IL-10 production while suppressing IL-6, IL-12, and TNF secretion. The compound's effects on cytokine production are mediated through its inhibition of SIK kinases and subsequent modulation of the CREB-regulated transcriptional co-activator (CRTC) pathway. |
| ln Vivo |
In vivo, MRT-199665 has been studied for its immunomodulatory effects. As a SIK inhibitor, it is expected to increase IL-10 production and suppress pro-inflammatory cytokines in vivo. The compound has been used to study the role of SIK kinases in inflammatory diseases. Specific in vivo efficacy data are limited in the published literature. MRT-199665 is a research tool for studying the role of MARK, SIK, and AMPK kinases in inflammation, metabolism, and cancer.
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| Enzyme Assay |
In vitro kinase assays for MRT-199665 typically involve measuring the inhibition of recombinant MARK, SIK, or AMPK kinases using radioisotopic or fluorescence-based methods. A typical protocol: recombinant kinase is incubated with varying concentrations of MRT-199665 (0.01 nM to 10 μM) in kinase assay buffer containing ATP and a peptide substrate. The reaction is incubated for 30-60 minutes at 30°C. Phosphorylated substrate is detected by scintillation proximity assay (SPA), time-resolved fluorescence resonance energy transfer (TR-FRET), or by Western blot using phospho-specific antibodies. IC₅₀ values are calculated from inhibition curves. Positive controls include staurosporine or other pan-kinase inhibitors.
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| Cell Assay |
Western Blot Analysis[2]
Cell Types: OCI-AML2 and MOLM-13 Cell Tested Concentrations: 10, 100, 500, and 1000 nM Incubation Duration: 12 Experimental Results: Compared to untreated cells, 10 nM resulted in total MEF2C and pS222 dose-dependently diminished MEF2C, resulting in a >40% reduction in MEF2C phosphorylation. Cell proliferation assay [2] Cell Types: human AML cell lines OCI-AML2, MV4-11, MOLM-13 and Kasumi-1 with endogenous MEF2C phosphorylation; human AML cell lines NB-4, HEL, HL lacking MEF2C -60 and U937 Tested Concentrations: 1 nM, 10 nM, 100n M, 1 μM, 10 μM, 100μM Incubation Duration: 48 hrs (hours) Experimental Results: Human AML cell lines with endogenous MEF2C phosphorylation vs. cell lines lacking MEF2C (NB- 4. Compared with HEL, HL-60 and U937), (OCI-AML2, MV4-11, MOLM-13 and Kasumi-1) are more sensitive, with an average IC50 of 26±13 and an average IC50 of 990 and ±29 nM respectively. . In vitro cell-based assays for MRT-199665 are performed using immune cells such as macrophages or dendritic cells. A typical protocol: cells are seeded in 96-well plates and treated with MRT-199665 at concentrations ranging from 0.01 to 10 μM for 1-2 hours. Cells are then stimulated with LPS (100 ng/mL) for 4-24 hours. Cytokine production (IL-10, IL-6, IL-12, TNF) is measured in the culture supernatant by ELISA. IL-10 and Nur77 mRNA levels are measured by qRT-PCR. Cell viability is assessed using MTT assays. Each condition is tested in triplicate, and experiments are repeated at least three times. |
| Animal Protocol |
In vivo animal studies for MRT-199665 are conducted in mouse models of inflammation or sepsis. A typical protocol: male C57BL/6 mice are administered MRT-199665 via intraperitoneal injection at doses of 1-30 mg/kg, 1-2 hours before LPS challenge (10-20 mg/kg, IP). Blood is collected 2-6 hours after LPS challenge, and cytokine levels (IL-10, IL-6, TNF) are measured in plasma by ELISA. Tissue samples (e.g., spleen, liver) are harvested for qRT-PCR analysis of cytokine mRNA. Efficacy is assessed by comparing cytokine levels between treatment and vehicle control groups.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of MRT-199665 have been partially characterized. The compound is soluble in DMSO. Its plasma half-life, volume of distribution, protein binding, and oral bioavailability have been evaluated in preclinical models. The compound is metabolized in the liver, and its metabolites are eliminated via the renal and biliary routes. Specific pharmacokinetic parameters are available from preclinical study reports. The compound should be stored at -20°C for long-term stability.
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| Toxicity/Toxicokinetics |
Toxicological data for MRT-199665 are limited to preclinical studies. The compound has not undergone formal toxicology testing for regulatory purposes. Standard laboratory safety precautions should be followed when handling MRT-199665: use of personal protective equipment (gloves, safety goggles, lab coat) and handling in a well-ventilated fume hood. The compound should be stored at -20°C for long-term stability. No genotoxicity, carcinogenicity, or reproductive toxicity data are available. Researchers should consult the safety data sheet (SDS) before handling.
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| References |
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| Additional Infomation |
Additional information for MRT-199665: The compound has a CAS number of 1456858-57-3. Its molecular formula is C₂₈H₃₁N₅O₂ and molecular weight is 469.58 g/mol. It is a potent and ATP-competitive, selective MARK/SIK/AMPK inhibitor. IC₅₀s: MARK1/2/3/14 = 2/2/3/2 nM, AMPKα1/α2 = 10/10 nM, SIK1/2/3 = 110/12/43 nM. It increases IL-10 and suppresses IL-6, IL-12, and TNF. It is for research use only and is not approved for clinical applications. No FDA approvals exist.
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| Molecular Formula |
C18H19N3O4S
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| Molecular Weight |
373.426162958145
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| Exact Mass |
469.247
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| CAS # |
1456858-57-3
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| PubChem CID |
71725150
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
4.3
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
35
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| Complexity |
770
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| Defined Atom Stereocenter Count |
1
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| SMILES |
S(C)(C1=NC=C2C(=N1)N(C(C2(C)C)=O)[C@@H]1C2C=CC=C(C=2CC1)O)(=O)=O
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| InChi Key |
LZUYOYKXEXENTQ-ZDUSSCGKSA-N
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| InChi Code |
InChI=1S/C18H19N3O4S/c1-18(2)12-9-19-17(26(3,24)25)20-15(12)21(16(18)23)13-8-7-11-10(13)5-4-6-14(11)22/h4-6,9,13,22H,7-8H2,1-3H3/t13-/m0/s1
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| Chemical Name |
7-[(1S)-4-Hydroxy-2,3-dihydro-1H-inden-1-yl]-5,5-dimethyl-2-(methylsulfonyl)-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one
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| Synonyms |
MRT199665 MRT-199665 MRT 199665
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~266.20 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.43 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 (4.43 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.43 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6779 mL | 13.3894 mL | 26.7788 mL | |
| 5 mM | 0.5356 mL | 2.6779 mL | 5.3558 mL | |
| 10 mM | 0.2678 mL | 1.3389 mL | 2.6779 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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