IUPAC/Chemical Name
(2E)-4-[Hydroxy[4-[[[4-[[[(1-naphthalenylamino)carbonyl]oxy]methyl]phenyl]methyl]-amino]butyl]amino]-4-oxo-2-butenoic acid methyl ester
InChi Key
MUJOCHRZXRZONW-FOCLMDBBSA-N
InChi Code
InChI=1S/C28H31N3O6/c1-36-27(33)16-15-26(32)31(35)18-5-4-17-29-19-21-11-13-22(14-12-21)20-37-28(34)30-25-10-6-8-23-7-2-3-9-24(23)25/h2-3,6-16,29,35H,4-5,17-20H2,1H3,(H,30,34)/b16-15+
SMILES Code
O=C(OC)/C=C/C(N(O)CCCCNCC1=CC=C(COC(NC2=C3C=CC=CC3=CC=C2)=O)C=C1)=O
Purity
>98% (or refer to the Certificate of Analysis)
Shipping Condition
Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition
Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility
Soluble in DMSO
Shelf Life
>2 years if stored properly
Drug Formulation
This drug may be formulated in DMSO
Stock Solution Storage
0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code
2934.99.9001
Biological target:
Methylstat is an inhibitor of the Jumonji C domain-containing histone trimethyl demethylases (JHDMs).
In vitro activity:
Methylstat inhibited KYSE150 cell growth with a half maximal growth inhibitory concentration (GI50) at approximately 5.1 μM (Figure 3a). Western blotting experiments showed that methylstat induces hypermethylation of lysine residues including H3K4, H3K9, H3K27, and H3K36 at almost all methylation states investigated in a concentration-dependent manner (Figure 3c). Similar results were also observed in human breast adenocarcinoma MCF7 cells (SI, Figure S3). Of substantial interest is the observation that methylstat also induces increased cellular levels of H3K79me3 and H4K20me3.
Reference: J Am Chem Soc. 2011 Jun 22;133(24):9451-6. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3133600/
In vivo activity:
Consistent with biological differences ex vivo, RCC (right-sided colorectal cancer) displayed more aggressive tumor growth than LCC-PDO in NSG mice. Strikingly, efficient re-compaction of chromatin in RCC organoids in vivo via administration of methylstat after reaching similar tumor size (50 mm3) resulted in reduced RCC tumor load, whereas this study did not observe changes in subcutaneous tumor growth of LCC in response to methylstat (Figure 1D and Supplementary Figure 1G). These data suggest that H4K20me3-mediated heterochromatin maintenance impacts RCC tumorigenesis.
Reference: Gastroenterology. 2023 Feb;164(2):214-227. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9889219/
Preparing Stock Solutions
The following data is based on the
product
molecular weight
505.57
Batch specific molecular weights may vary
from batch to batch
due to the degree of hydration, which will
affect the solvent
volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass |
1 mg |
5 mg |
10 mg |
| 1 mM |
1.15 mL |
5.76 mL |
11.51 mL |
| 5 mM |
0.23 mL |
1.15 mL |
2.3 mL |
| 10 mM |
0.12 mL |
0.58 mL |
1.15 mL |
| 50 mM |
0.02 mL |
0.12 mL |
0.23 mL |
Formulation protocol:
1. Koca D, Hastar N, Engür S, Kiraz Y, Ulu GT, Çekdemir D, Baran Y. Therapeutic Potentials of Inhibition of Jumonji C Domain-containing Demethylases in Acute Myeloid Leukemia. Turk J Haematol. 2020 Feb 20;37(1):5-12. doi: 10.4274/tjh.galenos.2019.2019.0083. Epub 2019 Dec 13. PMID: 31833715; PMCID: PMC7057756.
2. Luo X, Liu Y, Kubicek S, Myllyharju J, Tumber A, Ng S, Che KH, Podoll J, Heightman TD, Oppermann U, Schreiber SL, Wang X. A selective inhibitor and probe of the cellular functions of Jumonji C domain-containing histone demethylases. J Am Chem Soc. 2011 Jun 22;133(24):9451-6. doi: 10.1021/ja201597b. Epub 2011 May 31. PMID: 21585201; PMCID: PMC3133600.
3. Boonsanay V, Mosa MH, Looso M, Weichenhan D, Ceteci F, Pudelko L, Lechel A, Michel CS, Künne C, Farin HF, Plass C, Greten FR. Loss of SUV420H2-Dependent Chromatin Compaction Drives Right-Sided Colon Cancer Progression. Gastroenterology. 2023 Feb;164(2):214-227. doi: 10.1053/j.gastro.2022.10.036. Epub 2022 Nov 17. PMID: 36402192; PMCID: PMC9889219.
In vitro protocol:
1. Koca D, Hastar N, Engür S, Kiraz Y, Ulu GT, Çekdemir D, Baran Y. Therapeutic Potentials of Inhibition of Jumonji C Domain-containing Demethylases in Acute Myeloid Leukemia. Turk J Haematol. 2020 Feb 20;37(1):5-12. doi: 10.4274/tjh.galenos.2019.2019.0083. Epub 2019 Dec 13. PMID: 31833715; PMCID: PMC7057756.
2. Luo X, Liu Y, Kubicek S, Myllyharju J, Tumber A, Ng S, Che KH, Podoll J, Heightman TD, Oppermann U, Schreiber SL, Wang X. A selective inhibitor and probe of the cellular functions of Jumonji C domain-containing histone demethylases. J Am Chem Soc. 2011 Jun 22;133(24):9451-6. doi: 10.1021/ja201597b. Epub 2011 May 31. PMID: 21585201; PMCID: PMC3133600.
In vivo protocol:
1. Boonsanay V, Mosa MH, Looso M, Weichenhan D, Ceteci F, Pudelko L, Lechel A, Michel CS, Künne C, Farin HF, Plass C, Greten FR. Loss of SUV420H2-Dependent Chromatin Compaction Drives Right-Sided Colon Cancer Progression. Gastroenterology. 2023 Feb;164(2):214-227. doi: 10.1053/j.gastro.2022.10.036. Epub 2022 Nov 17. PMID: 36402192; PMCID: PMC9889219.
1: Lei M, Zhang H, Julian R, Tang K, Xie S, Zhu JK. Regulatory link between DNA methylation and active demethylation in Arabidopsis. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3553-7. doi: 10.1073/pnas.1502279112. Epub 2015 Mar 2. PubMed PMID: 25733903; PubMed Central PMCID: PMC4371987.
2: Xu W, Podoll JD, Dong X, Tumber A, Oppermann U, Wang X. Quantitative analysis of histone demethylase probes using fluorescence polarization. J Med Chem. 2013 Jun 27;56(12):5198-202. doi: 10.1021/jm3018628. Epub 2013 Jun 13. PubMed PMID: 23725560; PubMed Central PMCID: PMC3724763.
3: Luo X, Liu Y, Kubicek S, Myllyharju J, Tumber A, Ng S, Che KH, Podoll J, Heightman TD, Oppermann U, Schreiber SL, Wang X. A selective inhibitor and probe of the cellular functions of Jumonji C domain-containing histone demethylases. J Am Chem Soc. 2011 Jun 22;133(24):9451-6. doi: 10.1021/ja201597b. Epub 2011 May 31. PubMed PMID: 21585201; PubMed Central PMCID: PMC3133600.
