Raymond F. Schinazi. 3. 1. University ..... [12] Schinazi RF, Lloyd RJ, Nguyen M-H, Cannon DL, McMillan, Ilksoy N, Chu CK, Liotta DC, Bazmi. HZ & Mellors JW.
Mutations in RT and Protease
Mutations in HIV-1 Reverse Transcriptase and Protease Associated with Drug Resistance John W. Mellors,1 Brendan A. Larder,2 Raymond F. Schinazi3 1
University of Pittsburgh/VAMC, Graduate School of Public Health, 403 Parran Hall, 130 Desoto St, Pittsburgh, PA 15261, 2 The Wellcome Foundation, Ltd., Langley Court, Beckenham, Kent BR3 3BS, UK. 3 Emory University/VAMC, 1670 Clairmont Road, Decatur, GA 30033. INTRODUCTION The knowledge of HIV-1 drug resistance continues to increase at an exponential rate. This is particularly true for resistance to HIV-1 protease inhibitors, for which the first resistant variants were described less than two years ago (Otto et al., PNAS 90:7453–7457, 1993). Now resistance to virtually all classes of protease inhibitors has been documented. To keep our readership informed of newly described mutations, we have updated the tables below, which first appeared in IAVN in May 1994 (Vol 2; No 5). Following the tables is a list of abbreviations used. We hope that the tables’ revised content will be useful to virologists and clinicians active in the field. We urge all investigators to provide additions or amendments of the table to any of the authors. Data formatted as in the table with an appropriate reference (abstract, manuscripts or paper) would be welcomed. ACKNOWLEDGMENTS The authors gratefully acknowledge Margaret Tisdale, Michael J. Otto, Martin L. Bryant, Daniel W. Norbeck, Pin-Fang Lin and Emilio A. Emini for contributing essential information to the table.
III--93 NOV 95
Mutations in RT and Protease NUCLEOSIDE RT INHIBITORS Compound Amino Acid Change AZT
ddI
ddC
D4T
3TC or (-)-FTC Foscarnet
1592U89
Codon Change
In In Comments Vitro Vivo
Confirmed by Site-directed Mutagenesis
M41L D67N K70R T215Y T215F K219Q K219E
ATG to TTG GAC to AAC AAA to AGA ACC to TAC ACC to TTC AAA to CAA AAA to GAA
n.d Yes Yes Yes n.d. n.d. n.d.
Yes Yes Yes Yes Yes Yes No
M41L: 4-fold resistance; M41L/T215Y: 60- to 70-fold; K67N/K70R/T215Y/ K219Q: 120-fold; M41L/K67N/ K70R/T215Y: 180-fold. Effect of T215Y is reversed by a ddI resistance mutation (L74V), NNRTI mutations (L100I; Y181C) or a (-)-FTC/3TC mutation (M184V)
Yes Yes Yes Yes Yes Yes Yes
L74V
TTA to GTA
No
Yes
V75T
GTA to ACA
Yes
No
M184V
ATG to GTG
Yes
Yes
5- to 10-fold resistance; cross-resistance Yes to ddC; can reverse the effect of the T215Y AZT resistance mutation observed with D4T selection; crossYes resistance to ddI/ddC 3- to 4-fold resistance; cross-resistance Yes to ddC
K65R
AAA to AGA
Yes
Yes
T69D L74V V75T M184V Y215C
ACT to GAT TTA to GTA GTA to ACA ATG to GTG TTC to TGC
No
Yes
No
Yes
I50T V75T
ATT to ACT GTA to ACA
Yes Yes
M184V M184I
ATG to GTG or GTA ATG to ATA
W88S E89G
Refs.
[1, 2, 3]
[4] [5] [6]
4- to 10-fold resistance; observed in pts on receving ddI or ddC therapy 5-fold resistance observed with ddI therapy observed with D4T selection in vitro observed with ddI, 3TC therapy 4-fold resistance; arises on background of T215YAZT resistance mutation
Yes
[7, 8]
Yes Yes Yes Yes Yes
[9] [2] [4] [6] [10]
? Yes
30-fold resistance 7-fold resistance; cross-resistance to ddI, ddC, d4C, and (-)-FTC
Yes Yes
[11] [5]
Yes
Yes
Yes
[12–14]
Yes
Yes
100-fold resistance; M184V and M184I can suppress effects of AZT resistance mutations
Yes
[12–14]
TGG to TCG GAA to GGA
No Yes
Yes No
Yes Yes
[15] [16]
E89K L92I S156A Q161L H208Y
GAA to GGA TTA to ATA TCA to GCA CAA to CTA CAT to TAT
Yes Yes Yes Yes Yes
No No No Yes Yes
4-fold resistance isolated by screening RT clones for ddGTP resistance; 14-fold viral resistance E89K and L92I cause increased susceptibility to AZT and HIV-1 specific RTI 10-fold resistance 2-fold resistance Q161L + H208Y cause increased susceptibiltity to AZT (100-fold), nevirapine (20-fold) and TIBO R82150 (30-fold)
Yes Yes
[17] [17] [17] [15] [15]
K65R L74V Y115F M184V
AAA to AGA TTA to GTA TAT to TTT ATG to GTG
Yes Yes Yes Yes
No No No No
>
3-fold resistance Yes 4-fold resistance Yes 2-fold resistance Yes 3-fold resistance Yes K65R/M184V: 8-fold resistance L74V/M184V: 9-fold resistance L74V/Y115F/M184V: 11-fold resistance
III--94 NOV 95
[18]
Mutations in RT and Protease HIV-1-SPECIFIC RT INHIBITORS Compound
Amino Acid Change
Codon Change
In In Comments Vitro Vivo
Nevirapine
A98G L100I K103N V106A
GCA to GGA TTA to ATA AAA to AAC GTA to GCA
No No No Yes
Yes Yes Yes Yes
V108I Y181C
GTA to ATA TAT to TGT
No Yes
Yes Yes
Y181I Y188C G190A
TGT to ATT TAT to TGT GGA to GCA
No No No
Yes Yes Yes
TIBO R82150 L100I
TTA to ATA
Yes
?
TIBO R82913 L100I K103N V106A E138K Y181C Y188H Y188L
TTA to ATA AAA to AAC GTA to GCA GAG to AAG TAT to TGT TAT to CAT TAT to TTA
Yes Yes Yes Yes Yes Yes No
? ? ? ? ? ? Yes
L0 697,593
K103N Y181C
AAA to AAC TAT to TGT
Yes Yes
? ?
L0 697,661
A98G L100I K101E K103N K103Q V108I V179D V179E Y181C
GCA to GGA TTA to ATA AAA to GAA AAA to AAC AAA to CAA GTA to GCA GTT to GAT GTT to GAG TAT to TGT
No Yes No Yes No Yes No No Yes
Yes No Yes Yes Yes Yes Yes Yes Yes
BHAP U-90152 (delaviridine)
P236L
CCT to CTT
Yes
?
BHAP U-87201 (ateviridine)
K101E K103N Y181C Y188H E233V P236L K238T
AAA to GAA AAA to AAC TAT to TGT TAT to CAT GAA to GTA CCT to CTT AAA to ACA
No No No No No Yes No
Yes Yes Yes Yes Yes No Yes
L100I V106A Y181C Y181I
TTA to ATA GTA to GCA TAT to TGT TGT to ATT
Yes Yes Yes Yes
? ? ? Yes
Y188C Y181C Y106A
TAT to TGT TAT to TGT GTA to GCA
Yes Yes Yes
? ? ?
BHAP U 88204
HEPT E-EBU E-EBU-dM
Confirmed by Site-directed Mutagenesis
Ref.
Yes Yes Yes Yes
[19] [20] [20] [19–22]
Yes
[20] [19, 23–25]
100-fold resistance; no effect on AZT resistance
>100-fold resistance; cross-resistanceYes
to other NNRTI; can suppress effects of AZT resistance mutations High-level resistance
Yes Yes
>
100-fold resistance; can reverse Yes effects of AZT resistance mutations Yes 100-fold resistance Yes 100-fold resistance Yes
[26] [20] [19] [27–29]
Yes Yes Yes
[21] [22] [21] [28] [21] [28] [30]
20-fold resistance 100-fold resistance
>
Yes Yes
[23] [23]
8-fold resistance 2-fold resistance 8-fold resistance 8-fold resistance 8-fold resistance 4-fold resistance 4-fold resistance 8-fold resistance 30-fold resistance
Yes Yes Yes Yes Yes Yes Yes Yes Yes
[31] [31] [31] [31] [32] [31] [31] [31] [31]
Yes
[33]
> >100-fold resistance
>
P236L sensitizes RT 10-fold to Nevirapine, TIBO R82193, and L0 697,661 K103N and Y181C observed with U87201 monotherapy; K101E, Y188H, E233Y and K238T observed with U87201/AZT combination therapy
[34]
Yes
Yes Yes High-level resistance to BHAP, Nevirapine, and TIBO. Observed in one Nevirapine-treated patient
III--95 NOV 95
[34] [34] [34] [33] [34] [22, 35] [35] [35] [36]
[37] [37] [37]
Mutations in RT and Protease HIV-1-SPECIFIC RT INHIBITORS (cont.) Compound
Amino Acid Change
Codon Change
In Vitro
In Comments Vivo
E-EPU and E-EPSeU
Y181C Y188C
TAT to TGT TAT to TGT
Yes Yes
? ?
-APA R18893
Y181C
TAT to TGT
Yes
?
S-2720
G190E
GGA to GAA
Yes
?
TSAO
E138K
GAG to AAG
Yes
?
BM+51.0836
Y181C
TAT to TGT
Yes
?
Confirmed by Site-directed Mutagenesis
Ref.
Y188C is the predominant mutation Yes for E-EPSeU; Y188C confers greater Yes resistance to E-EPSeU/E-EPU than Y181C Yes
[38] [38]
Mutation decreases RT activity and viral replication competency >100-fold resistance
Yes
[40]
Yes
[41, 42]
Yes
[43]
III--96 NOV 95
[39]
Mutations in RT and Protease PROTEASE INHIBITORS Compound
A-77003
Amino Acid Change
Codon Change
In Vitro
In Comments Vivo
R8Q R8K V32I M46I
CGA to CGA to GTA to ATG to
CAA AAA ATA ATA
Yes Yes Yes Yes
? ? ? ?
M46L M46F G48V V82I
ATG to TTG ATG to TTC GGG to GTG GTC to ATC
Yes Yes Yes Yes
? ? ? ?
V82A I82T
GTC to GCC ATC to ACC
Yes Yes
? ?
A75925
V32I
GTA to ATA
Yes
?
40-fold viral resistance
ABT-538
V82F I84V
GTC to TTC ATA to GTA
Yes Yes
? ?
V82F/I84V: 8- to 10-fold viral resistance M46I/L63P/A71V/V82F/I84V: 27-fold resistance
BILA 1906 BS
V32I M46L A71V I84V
GTA to ATA ATG to TTG GCT to GTT ATA to GTA
Yes Yes Yes Yes
? ? ?
A71T V82A L10R M46I L63P V82T I84V
GCT to ACT CTC? to CGC? ATG to ATA CTC? to CCC? GTC to ACC ATA to CTA
Yes Yes No No No No No
? ? Yes Yes Yes Yes Yes
V32I M46I A71V V82A
GTA to ATA ATG to ATA GCT to GTT GTC to GCC
Yes Yes Yes Yes
? ? ? ?
P9941
V82A
GTC to GCC
Yes
?
Ro 31-8959
G48V I84V L90M
GGG to GTG ATA to GTA TTG to ATG
Yes Yes Yes
Yes ? Yes
BMS 186,318 L-735, 524
10-fold viral resistance 10-fold viral resistance 7-fold enzyme resistance No affect on susceptibility but restores replication kinetics of R8Q mutant 2–3-fold enzyme resistance 4-fold enzymee resistance R8K/M46I/G48V: 20-fold viral resistance no resistance alone but V32I and V82I are synergistic mutations yielding 20-fold enzyme resistance rare; seen with M46F G48V/I82Tcombined produce 100-fold viral resistance
Confirmed by Site-directed Mutagenesis
Refs.
Yes Yes Yes Yes
[44, 45] [44] [45] [44]
Yes Yes
[45] [45] [46] [45]
Yes
[47] [47] [48] Yes Yes
[49] [50]
V32I/A71V: 3-fold viral resistance V32I/A71V/M46I/I84V: 5-fold V32I/A71V/M46I/I84V: 1000-fold (mutation also detected in p6/p7 cleavage site) A71T/V82A: 15-fold viral resistance; 4-fold cross resistance to A77003 M46I/L63P/V82T: 4-fold viral resistance L10R/M46I/L63P/V82T: 4-fold viral resistance L10R/M46I/L63P/V82T/I84V: 8-fold viral resistance; cross resistance to XM-323 (15fold], A-80987 (4-fold), Ro-31-8959 (8-fold), VX-478 (8-fold), SC-52151 (8-fold) V32I/M46L/V82A: 3-fold viral resistance V32I/M46L/A71V/V82A: 14-fold viral resistance
Yes Yes Yes
[51]
6–8-fold resistance
Yes
[53]
Yes
[54] [46]
[52]
[46]
G48V/L90M combined yield
>100-fold enzyme resistance, but
RPI-312
I84V
ATA to GTA
Yes
?
double mutant rare in vivo; L90M most common in vivo G48V/I84V/L90M: 90-fold viral resistance 5-fold viral resistance
SC-52151
L24V G48V A71V V75I P81T
TTA to GTA GGG to GTG GCT to GTT GTA to ATA CCT to ACT
Yes Yes Yes Yes Yes
? ? ? ? ?
N88D: 10-fold viral resistance G48V/V82A: 15-fold viral resistance G48V/A71V/V75I/P81T: 20- to 30-fold A71V/V75I/P81T: 30-fold viral resistance L24V/G48V/A71V/V75I/P81T: 1000-fold
III--97 NOV 95
[54] [46] Yes
[55] [56, 57]
Mutations in RT and Protease PROTEASE INHIBITORS (cont.) Compound Amino Acid Change
Codon Change
In In Comments Vitro Vivo
Confirmed by Site-directed Mutagenesis
Refs.
V82A N88D
GTC to GCC AAT to GAT
Yes Yes
? ?
viral resistance; some cross resistance to SC55389a and Ro31-8959, but not to L-735,524
SC-55389
L10F N88S
CTC to CGC AAT to AGT
Yes Yes
? ?
N88S seen alone with L10F; 10- to 20-fold viral resistance
VB 11,328
L10F M46I I47V I50V I84V
CTC to GGC ATG to ATA ATA to CTA ATT to GTT ATA to GTA
Yes Yes Yes Yes Yes
? ? ? ? ?
L10F/I84V: 10-fold viral resistance I50V/M46I/I47V: 20-fold viral resiestance
Yes
[58] [46, 58]
3-fold viral resistance
Yes
[46]
L10F K45I G48V V82A
CTC to CGC AAA to ATA GGG to GTG GTC to GCC
L10F/V82A: 2.0-fold viral resistance; L10F/K45I/I84V: 50-fold
Yes
Yes Yes
? ?
Yes Yes
[59] [46] [59] [59]
V82I V82F I84V
GTC to ATC GTC to TTC ATA to GTA
Yes Yes Yes
? ? ?
Yes Yes Yes
[59] [59] [46, 59]
L97V
TTA to GTA
Yes
?
Yes
[59]
XM323
V82A/M46L: 7-fold resistance V82A/M46L/L97V: 11-fold resistance 2-fold viral resistance see below 12-fold resistance alone; V82F/I84V: 92fold resistance; cross resistant to P9941, but not A77003 or Ro 31-8959 no resistance alone; V82A/L97V: 3.0 fold resistance
