Damanhour University

Faculty of Pharmacy
Pharmaceutical Chemistry Department

Identification of Navoximod as selective

indoleamine-2,3-dioxygenase inhibitor

Prepared by: Alaa Osama Soliman Zayed

Level: Fifth year
ID: 5061
Under supervision of: Dr. Shaymaa Kassab
1. Target identification
Indoleamine-2,3-dioxygenase 1 (IDO1).
2. Target validation
The complex crystal structure of the hIDO1 with 4-phenylimidazole (4-
PI). (PDB code: 2D0T)
Besides, the X-ray image of the complex showed that N3-4-PI
coordinates with Fe-heme.
3. Lead identification
The discovery of Navoximod (GDC-0919) started in 1989 with the
identification of 4-phenylimidazole (4-PI), having a drug-like profile with
a weak non-competitive hIDO1 inhibiting activity (IC50 = 28 µM)[24,
25].
 4. Lead optimization
The structure-activity relationship studies disclosed by Kumar et al. [27] tried to improve the
activity against hIDO1, highlighting some considered fundamentals in the modification axes
of the hit [23]:
.1introduction of 2`-OH to 4-PI [Figure 2 (2)] improves the activity 16 times (IC50
= 1.7 µM) the original ligand 4-PI (IC50 = 28 µM) but 2',6`-dihydroxy-4-PI gives insignificant
effect,
.2phenyl ring accommodates small-size groups and the activity significantly increases with
the introduction of 3`-F,5`-Cl to 2`-OH-4-PI [Figure (3)] to exhibit a 5-fold increase in the
activity (IC50 = 0.3 µM),
.3the short half lifetime (t½=1h) of 2`-OH, 3`-F, 5`-Cl-4-PI [Figure 2 (3)] is attributable to the
metabolic-labile phenolic OH group,
.4improvement of the ligand biochemical activity, and PK profile without compromising the
cellular activity is challengeable,
.5 2`-OH, 6`-cyclohexylethoxy-4-PI [Figure 2 (4)] gives equipotent compound
(IC50 = 1.7 µM) to 2`-OH-4-PI [Figure 2 (2)] but weak cellular activity (EC50 = 80
µM).
6. Identification of the freely rotating 2`-cyclohexylethoxy-4-PI [Figure 2 (4)] having
improved anti-enzymatic activity (IC50 = 1.7 µM) motivated the group to introduce the
ethylcyclohexyl group at carbon 5 of imidazoisoindole and enhanced the activity of the
racemic mixture.
7. The introduction of 11-OH imparts 4-different diastereomers that are difficult to
isolate. So, the biochemical assay was applicable for the diastereomeric mixture to show
a 2-fold increase in the biochemical activity (IC50 = 0.060 µM) consistently with a 13-
fold improvement in the cellular potency (EC50 = 0.083 µM).
8. The profile of the resulting derivative [Figure 2 (9)] shows the best balance of potency,
metabolic stability, and on-target activity. Eventually, the S,R-trans stereoisomer [Figure
2 (Navoximod)] is the best to show potential activity at (hIDO1 IC50 = 0.028 µM), cellular
potency at (IC50 = 0.075 µM), CYP 3A4 inhibiting activity at (IC50 = 5.70 µM) and hepatic
clearance is very acceptable.
 5. Docking
The complex crystal structure of hIDO1 with Navoximod
(PDB code: 6O3I) [71] revealed the binding pattern of the
inhibitor to the protein binding site. It emphasized the
occupancy of the imidazoisoindole core structure to the
hydrophobic space of the A-pocket to look coated with the
hydrophobic amino acid residues of Phe163, Phe164,
Val130, and Tyr126 (Figure 3). Moreover, Cys129-SH was
proximal to 6-fluorine of Navoximod without any
opportunity for fluorine-sulfur interaction [71]. The
interaction of the inhibitor extended to B-pocket via π-
stacking of the 15-hydroxycyclohexyl fragment with Phe226
and hydrogen bond with Ser235 (Figure 9). 11-OH formed
H-bond with 7-propionate-Heme at a reasonable distance.
6. Preclinical trials
Not mentioned.

7. Clinical trials
Phase I clinical trials of Navoximod combined with the checkpoint
inhibitor, Atezolizumab [85] on patients with advanced solid tumors,
showed acceptable safety. However, there is no clear evidence of the
therapeutic benefit [91]. In another trial, it is well-tolerated at a high
dose (800 mg) and gives a stable disease response [92].