ML147 : HPGD (15-hydroxyprostaglandin dehydrogenase [NAD(+)]) Inhibitor
ML147
Target Name
15-hydroxyprostaglandin dehydrogenase [NAD(+)]
Target Alias
HPGD
Target Class
Dehydrogenase
Mechanism of Action
Inhibitor of HPGD
Biological / Disease Relevance
Pathways of Prostaglandins, Biological function and intracellular interactions of 15-PGDH
In vitro activity
HPGD bioassay (IC50)In vitro activity
HPGD Thermal shift assay (delta Tm)Target Information
15-hydroxyprostaglandin dehydrogenase (15-PGDH; HPGD) is the key enzyme for the inactivation of prostaglandins, and thus regulates processes such as inflammation or proliferation. The anabolic pathways of prostaglandins are well-characterized, especially with respect to regulation of the cyclooxygenase (COX) enzymes. In comparison, little is known about downstream events, including functional interaction of prostaglandin-processing and metabolizing enzymes, as well as the function of prostaglandin receptors. ML147 (CID-3245059) is a potent and competitive HPGD inhibitor that is selective within the dehydrogenase family.
Properties
ML147
NCGC00044151
| Physical & chemical properties | ||||
|---|---|---|---|---|
| Molecular Weight | 345.22 g/mol | |||
| Molecular Formula | C14H9BrN4S | |||
| cLogP | 3.5 | |||
| PSA | 90.7 | |||
| Storage | ||||
| Solubility | ||||
| CAS Number | 355827-05-3 | |||
SMILES:
BrC1=CNC2=NC(SCC3=C(C#N)C=CC=C3)=NC2=C1
InChI:
InChI=1S/C14H9BrN4S/c15-11-5-12-13(17-7-11)19-14(18-12)20-8-10-4-2-1-3-9(10)6-16/h1-5,7H,8H2,(H,17,18,19)
InChIKey:
DEYMHFHACXKPOV-UHFFFAOYSA-N
Activity
Summary activity statement /
ML147 (SID 87550717; CID 3245059) is a potent and selective inhibitor of HPDG. This probe has been tested in > 459 assays, and thus far, have only been active in HPGD-related assays performed at NCGC. ML147 is also found inactive against a related dehydrogenases.
In vitro activity - Selectivity Assay
| ML147 (IC50) | |
|---|---|
|
HPGD bioassay |
0.141 uM |
|
ALDH1A bioassay |
> 57 uM |
|
HADH2 bioassay |
> 57 uM |
|
HSD17beta4 bioassay |
> 57 uM |
Summary /
Selectivity of the ML147 is determined. Two structurally related dehydrogenases were chosen as suitable anti-targets for characterizing selectivity of the probe series: HADH2 and HSD17β4. The third anti-target is an ALDH1A1 which shares some homology with HPGD. ML147 is observed to be > 2000 fold, > 1000 fold, and > 1000 fold selective towards HPGD vs ALDH1A, HADH2, and HSD17beta4 respectively.
In vitro activity - ADME Profiling
| ML147 | |
|---|---|
|
Aqueous (kinetic) solubility in PBS buffer (pH 7.4 @ 25C) |
3 uM |
|
Experimental LogD |
2.35 |
|
Buffer Stability (percent remaining after 48hr at 25C) |
100 % |
Summary /
ADME data shows that the ML147 is stable in buffer even after 48hrs.
In vitro - Thermal Shift Assay
Summary /
ML147 is observed to have a strong stabilization in the presence of presence of NAD+ but a weak stabilization in the presence of NADH (which is expressed as the shift in the transition midpoint temperature (ΔTm)). An IC50 of 26.4 nM was determined for the re-synthesized ML147 compound, while the purified re-purchased dry powder gave an IC50 of 35.6 nM.
Table 1. ML147: Effect of probe compound on stability and activity of HPGD.
In vitro activity - Mechanism of Action Studies
Summary /
A kinetic characterization in experiments with titration of the 15-PGDH substrate PGE2 and of co-subtrate NAD, respectively. The addition of the probe inhibitor at increasing concentrations to the substrate titration caused an increase in Km for PGE2, suggesting competition of the inhibitor with the substrate for ML147.
Figure 1. Mechanism-of-action studies of probe compound with respect to the substrate. Insets show the Michaelis-Menten graphs of 15-PGDH activity at increasing concentrations of PGE2, while the main graphs show plots of the data after Lineweaver and Burk. Dottedlines in both representations show fits to the Michaelis equation. Black represents no inhibitor, blue denotes 10nM probe compound, and red denotes 50nM probe compound.
References
- Probe Development Summary for Inhibitors of HPGD (15-Hydroxyprostaglandin Dehydrogenase)
- Jadhav A, Niesen FH, Schultz L, et al. Potent and selective inhibitors of NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (HPGD) 2010 Mar 4 [Updated 2011 Mar 11]. In: Probe Reports from the NIH Molecular Libraries Program [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2010