| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg | |||
| 250mg | |||
| Other Sizes |
Purity: ≥98%
| Targets |
EZH2 (wild‑type) – biochemical IC50 = 0.002 μM (2 nM) [1];
EZH2 (Y641N mutant) – IC50 = 0.0031 μM (3.1 nM) [1]; EZH1 – IC50 = 52 ± 11 nM [1] |
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| ln Vitro |
Treatment with EZH2 inhibitor CPI-1205 cause apoptosis in multiple myeloma and plasmacytoma cell models
Cell Assay: Previous study reported that the treatment with CPI-1205 caused apoptosis in multiple myeloma and plasmacytoma cell models. CPI-1205 also had a clean selectivity profile when tested against 30 other histone or DNA methyltransferases. In addition, CPI-1205 demonstrated modest selectivity when tested against enhancer of zeste homologue 1 (EZH1) that is a methyltransferase highly related to EZH2. CPI-1205 displayed potent biochemical inhibition of wild‑type EZH2 (IC50 = 0.002 μM) and the Y641N mutant (IC50 = 0.0031 μM) in a scintillation proximity assay (SPA). In the HeLa cellular mechanism of action (MOA) assay measuring global H3K27me3 levels, it showed an EC50 of 0.032 μM. Compound 13 demonstrated modest selectivity against EZH1 (IC50 = 52 ± 11 nM). Introduction of a single C663S mutation in EZH2 (equivalent to the EZH1 residue) reduced inhibitor potency, consistent with the observed EZH1/EZH2 selectivity. Compound 13 was tested against 30 other histone or DNA methyltransferases and exhibited a clean selectivity profile. In vitro profiling showed no time‑dependent inhibition (TDI) of CYP450 enzymes 1A2, 2C9, 2C8, 2D6, and 3A4. In a secondary pharmacology panel of 54 receptors, transporters, and ion channels at 10 μM, no target was inhibited more than 50%. The hERG binding IC50 was determined to be 21.3 μM, which is well above the predicted free concentration in patients. [1] |
| ln Vivo |
CPI-1205 shows excellent oral bioavailability. It was well-tolerated for repeat dosing as demonstrated by the absence of significant body weight loss. CPI-1205 shows relatively high clearance in both rats and dogs (3.19 L/h/kg and 1.41 L/h/kg, respectively) but demonstrates good oral bioavailability in both species(44.6% F in rats and 46.2% F in dogs). CPI-1205 was well-tolerated in the 28-day GLP toxicology studies, and any findings were reversible over the recovery period
In a KARPAS‑422 B‑cell lymphoma xenograft model (female CB‑17 SCID mice), oral administration of CPI-1205 at 160 mg/kg twice daily (BID) for 25 days resulted in significant tumor regression, with >97% tumor growth inhibition (TGI) relative to vehicle control. The compound was well‑tolerated, as evidenced by the absence of significant body weight loss. Pharmacodynamic analysis of tumor tissues harvested 1 hour after the last dose on day 25 revealed a 47% reduction in the H3K27me3/global H3 ratio compared to vehicle control. Plasma and tumor concentrations of 13 at 1 hour post last dose were 11,388 ng/mL (~22 μM) and 5,286 ng/g (~10 μM), respectively. [1] |
| Enzyme Assay |
The biochemical inhibitory activity of CPI-1205 was measured using a scintillation proximity assay (SPA) with recombinant PRC2 complex, a biotinylated oligonucleosome substrate, an H3K27me3 activator peptide, and [3H]‑S‑adenosyl‑L‑methionine ([3H]‑SAM). The reaction was incubated, and the signal was detected on a scintillation counter. IC50 values were calculated from concentration‑response curves, with each value representing the average of at least two independent determinations. For EZH1 inhibition, a similar SPA format was used with recombinant PRC2 containing EZH1. To assess the role of Cys663, a mutant EZH2 (C663S) was incorporated into reconstituted PRC2 and the potency of inhibitors was measured under the same assay conditions. [1]
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| Cell Assay |
The cellular potency of CPI-1205 was evaluated in a mechanism of action (MOA) assay using HeLa cells. Cells were treated with compound for a specified period, and then global levels of H3K27me3 were quantified. The EC50 value (0.032 μM) was determined from the concentration‑response curve as the concentration required to reduce H3K27me3 levels by 50% relative to control. Detailed assay conditions (cell seeding density, treatment duration, and detection method – e.g., ELISA or Western blot) are provided in the Supporting Information of the paper. [1]
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| Animal Protocol |
Formulated in 10% DMSO + 60% PEG400 + 30% ddH2O; 160 mg/kg orally twice daily; oral gavage CB-17 SCID mice
For in vivo pharmacokinetic studies, CPI-1205 was formulated for intravenous (iv) administration in a vehicle of 5:25:70 DMA/PEG400/20% SBECD, and for oral (po) administration in 0.5% methyl cellulose adjusted to pH 3–4. Male Balb/C mice received a single iv dose of 1 mg/kg and a single po dose of 100 mg/kg. For rat and dog PK, male Sprague‑Dawley rats received iv 1 mg/kg and po 25 mg/kg; male beagle dogs received iv 1 mg/kg and po 5 mg/kg. Blood samples were collected at various time points, and plasma concentrations were analyzed by LC‑MS/MS to determine PK parameters. For the KARPAS‑422 efficacy study, female CB‑17 SCID mice bearing established tumors were dosed orally with 13 at 160 mg/kg BID for 25 days. Tumor volumes and body weights were measured regularly. At the end of the study, tumors and plasma were collected 1 hour post last dose for PK/PD analysis. [1] |
| ADME/Pharmacokinetics |
Mouse PK (after iv 1 mg/kg and po 100 mg/kg): CL = 2.16 L/h/kg (40% of liver blood flow), t1/2 = 1.63 h, Vss = 1.4 L/kg, AUC0‑∞ = 88.88 μM·h (unbound = 1.87 μM·h), Cmax = 67.36 μM (unbound = 1.41 μM), oral bioavailability F = 100%, fraction unbound in plasma fu = 0.021 [1].
Rat PK (iv 1 mg/kg, po 25 mg/kg): CL = 3.19 L/h/kg (97% of liver blood flow), t1/2 = 0.65 h, Vd = 3.2 L/kg, AUC0‑∞ = 7.06 μM·h (unbound = 0.713), Cmax = 5.88 μM (unbound = 0.593), tmax = 0.50 h, F = 44.6%, fu = 0.098 [1]. Dog PK (iv 1 mg/kg, po 5 mg/kg): CL = 1.41 L/h/kg (76% of liver blood flow), t1/2 = 1.84 h, Vd = 1.6 L/kg, AUC0‑∞ = 3.19 μM·h (unbound = 0.309), Cmax = 1.24 μM (unbound = 0.120), tmax = 1.67 h, F = 46.2%, fu = 0.089 [1]. In vitro ADME: cLogP = 3.41, lipophilic efficiency (LiPE) = 4.2. Mouse liver microsome CLint = 131 μL/min/mg; rat = 71.3; dog = 52.4; human = 73.6. Plasma protein binding (% bound): mouse = 97.8%, rat = 83.1%, dog = 64.3%, human = 83.3%. CYP inhibition at 10 μM: CYP3A4 = 5.0%, CYP2D6 = 26.1%, CYP2C8 = 18.9%, CYP1A2 = 28.6%, CYP2C9 = 5.4% [1]. |
| Toxicity/Toxicokinetics |
A 28‑day GLP toxicology study was conducted in Sprague‑Dawley rats with once daily (QD) oral gavage doses of 100, 300, and 600 mg/kg, followed by a 4‑week recovery period. In beagle dogs, the compound was administered twice daily (BID) at 50, 150, and 500 mg/kg for 28 days, also with a 4‑week recovery. CPI-1205 was generally well‑tolerated, and all findings were reversible during the recovery period. No time‑dependent CYP inhibition was observed. The hERG binding IC50 was 21.3 μM, indicating a low risk of QT prolongation at therapeutic concentrations. At a concentration of 10 μM, the compound did not inhibit any target in a panel of 54 receptors, transporters, and ion channels by more than 50%. [1]
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| References |
J Med Chem.2016 Nov 10;59(21):9928-9941.
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| Additional Infomation |
Lilamestat is a potent and selective inhibitor of the histone methyltransferase EZH2. Lilamestat is an orally administered selective histone lysine methyltransferase EZH2 inhibitor with potential antitumor activity. After oral administration, lilamestat selectively inhibits the activity of both wild-type and mutant EZH2. Inhibition of EZH2 specifically prevents methylation of histone H3 at lysine 27 (H3K27). Reduced histone methylation alters gene expression patterns associated with cancer pathways and leads to reduced proliferation of EZH2-expressing cancer cells. EZH2 is a histone lysine methyltransferase (HMT) and the catalytic subunit of the polycomb repressor complex 2 (PRC2). It is overexpressed or mutated in various cancer cells and plays a crucial role in tumor cell proliferation; its expression is associated with tumorigenesis, development, stem cell self-renewal, migration, and angiogenesis.
CPI-1205 is a pyridone‑indole based EZH2 inhibitor that competes with S‑adenosyl‑L‑methionine (SAM). Co‑crystal structure of a related analogue (10) bound to human PRC2 (PDB code: 5LS6) reveals that the inhibitor binds at the interface of the EZH2 SET domain, the SAL region of the EZH2 N‑terminus, and EED. The binding induces a conformation in which the C‑terminus of the EZH2 SET domain occupies the histone binding groove, closing the lysine channel and making substrate binding incompatible. The compound is intended for the treatment of B‑cell lymphomas, including those harboring activating mutations in EZH2 (e.g., Y641N). Phase I clinical trial information: NCT02395601. The synthesis of CPI-1205 was achieved on >100 g scale via a palladium‑catalyzed intramolecular C–N arylation route. [1] |
| Molecular Formula |
C27H33F3N4O3
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|---|---|---|
| Molecular Weight |
518.57
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| Exact Mass |
518.25
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| Elemental Analysis |
C, 62.54; H, 6.41; F, 10.99; N, 10.80; O, 9.26
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| CAS # |
1621862-70-1
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| Related CAS # |
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| PubChem CID |
78320408
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| Appearance |
Solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
692.5±55.0 °C at 760 mmHg
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| Flash Point |
372.6±31.5 °C
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| Vapour Pressure |
0.0±2.2 mmHg at 25°C
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| Index of Refraction |
1.590
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| LogP |
3.94
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
37
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| Complexity |
926
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| Defined Atom Stereocenter Count |
1
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| SMILES |
FC(C([H])([H])N1C([H])([H])C([H])([H])C([H])(C([H])([H])C1([H])[H])[C@@]([H])(C([H])([H])[H])N1C2=C([H])C([H])=C([H])C([H])=C2C(C(N([H])C([H])([H])C2C(N([H])C(C([H])([H])[H])=C([H])C=2OC([H])([H])[H])=O)=O)=C1C([H])([H])[H])(F)F
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| InChi Key |
HPODOLXTMDHLLC-QGZVFWFLSA-N
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| InChi Code |
InChI=1S/C27H33F3N4O3/c1-16-13-23(37-4)21(25(35)32-16)14-31-26(36)24-18(3)34(22-8-6-5-7-20(22)24)17(2)19-9-11-33(12-10-19)15-27(28,29)30/h5-8,13,17,19H,9-12,14-15H2,1-4H3,(H,31,36)(H,32,35)/t17-/m1/s1
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| Chemical Name |
(R)-N-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-1-(1-(1-(2,2,2-trifluoroethyl)piperidin-4-yl)ethyl)-1H-indole-3-carboxamide
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| Synonyms |
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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 |
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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)
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| Solubility (In Vitro) |
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9284 mL | 9.6419 mL | 19.2838 mL | |
| 5 mM | 0.3857 mL | 1.9284 mL | 3.8568 mL | |
| 10 mM | 0.1928 mL | 0.9642 mL | 1.9284 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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