XVIII International AIDS Conference
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XVIII International AIDS Conference Novel HIV-1 Protease Inhibitors Containing bis- Tetrahydrofuran or a Novel Polycyclic Ligand, Pyranofuran 1 Depts of Hematology and Infectious Diseases, Kumamoto Univ; 2 Kumamoto Health Science Univ; 3 Depts. of Chem & Med Chem, Purdue Univ; 4 Experimental Retrovirology Section, NCI, NIH. Kazuhiko Ide 1 , M. Aoki 1,2 , Y. Koh 1 , M. Amano 1 , S. Kulkarni 3 , D.D. Anderson 3 , B. Chapsal 3 , R. Yedidi 4 , D. Das 4 , A.K. Ghosh 3 , and H. Mitsuya 1,4
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XVIII International AIDS Conference. Novel HIV-1 Protease Inhibitors Containing bis -Tetrahydrofuran or a Novel Polycyclic Ligand, Pyranofuran. Kazuhiko Ide 1 , M. Aoki 1,2 , Y. Koh 1 , M. Amano 1 , - PowerPoint PPT Presentation
Transcript of XVIII International AIDS Conference
XVIII International AIDS Conference
Novel HIV-1 Protease Inhibitors Containing bis-Tetrahydrofuran or a
Novel Polycyclic Ligand, Pyranofuran
1Depts of Hematology and Infectious Diseases, Kumamoto Univ;
2Kumamoto Health Science Univ; 3Depts. of Chem & Med Chem, Purdue Univ; 4Experimental Retrovirology Section, NCI, NIH.
Kazuhiko Ide1, M. Aoki1,2, Y. Koh1, M. Amano1, S. Kulkarni3, D.D. Anderson3, B. Chapsal3, R. Yedidi4,
D. Das4, A.K. Ghosh3, and H. Mitsuya1,4
BackgroundHAART made a major impact on the morbidity of HIV-1-infected individuals and significantly extended their survival.
The emergence of multi-drug-resistant viruses (HIV-1MDR) is still one of the major
concerns.
Continuous efforts are required to develop more potent and safer therapeutics with high genetic barrier.
OHN N
S
OH
OO O
NH2
O
O
HH
H
Darunavir
DRV possesses a dual antiviral activity, i.e., protease inhibition and protease dimerization inhibition. Koh & Mitsuya. J Biol Chem. 282:28709-20, 2007
The presence of bis-THF is important for the antiviral activity of DRV.
bis-THFbis-tetrahydrofuran
Koh, Ghosh & Mitsuya. Antimicrob Agents Chemother 47:3123-3129, 2003
OHN N
S
O
O
O
O
OH
OO
O
OHN N
S
O
O
O
O
OH
OO
O
GRL-0878 GRL-0888Major Isomer Minor Isomer
bis-THF bis-THF
5 56
6
( Cis ) ( Trans )
HN N
S
OH
OO
O
O
O
O
O
NH2
OHN N
S
O
OH
OH
O
O
O
O
O
GRL-1388 GRL-1398
polycyclic ligand polycyclic ligand(pyranofuran) (pyranofuran)
Drug EC50 (nM) CC50 (μM) * Selectivity Index
SQV 6.2 19.7 3,100
APV 19.6 >100 >5,100
ATV 5.1 27.6 5,400
DRV 3.3 >100 >30,300
0878 2.3 34.6 15,000
0888 5.1 33.6 6,200
1388 3.6 >100 >27,800
1398 0.2 37.8 186,800
*Selectivity index = CC50 / EC50
Four New Compounds Are Comparably orMore Potent against HIV Compared to DRV
Virus EC50 (nM)
ATV APV DRV 0878 0888 1388 1398
HIV-1WT (X4) 1.9 (1) 35.1 (1) 3.5 (1) 6.0 (1) 6.8 (1) 3.2 (1) 0.3 (1)
HIV-1MDR/B (X4) 228 (120) 328.8 (9) 19.4 (6) 59.1 (10) 82.3 (12) 4.7 (2) 4.8 (16)
HIV-1MDR/C (X4) 25.4 (13) 265.2 (8) 5.2 (1) 34.5 (6) 58.5 (9) 4.0 (1) 1.1 (4)
HIV-1MDR/MM (R5) 68.9 (36) 402.2 (11) 21.4 (6) 301.0 (50) 312.8 (46) 21.3 (7) 2.6 (9)
HIV-1MDR/EV (R5) 39.0 (21) >1,000 (28) 214.6 (61) >1,000 (167) >1,000 (147) 530.3 (166) 23.5 (78)
(fold change), blue ： 5-10 times, red ： >10 times
GRL-1398 Is More Potent against a Wide Spectrum of HIVMDR Isolates than DRV
VirusEC50 (nM)
DRV GRL-1388 GRL-1398
HIV-1WT 3.3 (1) 3.2 (1) 0.3 (1)
HIV-1DRVR
P10 29.1 (9) 24.6 (8) 3.3 (11)
HIV-1DRVR
P20 214 (65) 151 (47) 21.9 (73)
GRL-1398 Is Active against DRV-resistant HIV Variants
(Fold-change)
CFP
Monomer-monomer Interactions
Protease or Polyprotein Dimerization
FRET
YFP
RTYellow Fluorescent ProteinPR
F/P
Cyan Fluorescent ProteinPR AAAAA
AAAAA
Spacer
pHIV-1NL43sma
238 AA
RT
99 AAApa
FRET-HIV-1 Expression System
Protease orpolyproteinMonomer
Intermolecular FRET (Fluorescence Resonance Energy Transfer)-based
If CFPA/B ratios are < 1.0, there is no FRET or dimerization
Protease or polyprotein
Monomer
1 - 10 nm
1.0
0.9
0.8
0.7
0.6
0.5
1.1
1.2
1.3
1.4
1.5
0.86
CF
PA
/B r
atio
None GRL-1398GRL-1388 DRV
10 201
(μM)
1.12
1.02
0.83
1.10
0.96
0.88
0.92
0.850.81
GRL-1388 and -1398 have PDI activityD
imer
izat
ion
－D
imer
izat
ion
＋
10 201 10 201
p = 0.07
p <0.0001
p = 0.15
p = 0.0005
p = 0.0001
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 500.0001
0.001
0.01
0.1
1
10
Week
Dru
g c
on
ce
ntr
ati
on
(μ
M)
Cells used : MT4
A28S/M46I Confer Resistance to GRL-1398 on HIVNL4-3
GRL-1388
GRL-1398
DRV
L10F
L10FA28SM46I
L10F A28S M46I
V82I
L10FV32IM46L
APV
L10FV32IL33FM46LI54MA71V
Weeks
GRL-1398 Resists against Emergence of HIVDRV
R with a Mixture of 8 HIV-1MDR Isolates
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 290.0001
0.001
0.01
0.1
1
10
week
Dru
g c
on
ce
ntr
ati
on
(μ
M)
GRL-1388
GRL-1398
DRV
Cells used : MT4Weeks
L10IL33IM36I M46IL63PA71VV82AL90M
VAL82
VAL32
ILE84
ASP30
ASP29
ALA28
ASP29’
ASP30’
GLY49
GLY48
ILE47
ASP25
PRO81’
VAL82’
ILE84’
ILE50’
H2OGLY27’
ALA28’
GRL-1398 Has Tight Interactions with PR
Inhibitor H-bonds
DRV 11
1388 9
1398 13
InhibitorHydrophobicinteractions
DRV 26
1388 24
1398 28
SummaryGRL-1388 and -1398, containing a novel structure-based designed ligand, were comparably or more potent compared to DRV.
Although GRL-1398 had moderate PDI activity as compared to DRV, GRL-1398 exhibited more potent anti-HIV activity against DRV- resistant HIV variants.
ConclusionsGRL-1398 has activity against protease’s enzymatic activity and dimerization activity with high genetic barrier.
It is warranted that the compounds, especially GRL-1398, be further studied as potential therapeutics for HIVwt and HIV-1MDR infection.
Thank you for your attention
