Table 1.
Optimization of anti-HIV chemokines using phage display
| Molecule | N-terminal sequence |
Anti-HIV potency, pM | CCR5 signaling activity, % max | CCR5 sequestration, % control |
|---|---|---|---|---|
| 0–1–2–3–4–5–6–7–8–9 | ||||
| PSC-RANTES | PSC-P-Y-S-S-D-T-T-P | 25 | 100 | 75 |
| Library 1 | X-S-#-X-S-S-X-#-#-# | |||
| 1P1-RANTES | L-S-P-V-S-S-Q-S-S-A | 6,600 | BD | BD |
| 1P2-RANTES | F-S-P-L-S-S-Q-S-S-A | 1,600 | 94 | 68 |
| Second round of optimization | ||||
| Library 2 | X-X-P-X-X-X-Q-#-T-P | |||
| 2P1-RANTES | F-V-P-Q-S-G-Q-S-T-P | 7,900 | BD | BD |
| 2P2-RANTES | L-V-P-Q-P-G-Q-S-T-P | 17,000 | BD | BD |
| 2P3-RANTES | Q-G-P-P-L-M-Q-T-T-P | 650 | BD | BD |
| 2P4-RANTES | M-V-P-Q-S-G-Q-S-T-P | 18,000 | BD | BD |
| 2P5-RANTES | Q-G-P-P-M-M-Q-T-T-P | 5,000 | BD | BD |
| 2P6-RANTES | Q-G-P-P-G-G-Q-T-T-P | 13,000 | 37 | 56 |
| 2P7-RANTES | F-A-P-M-S-Q-Q-S-T-S | 1,400 | 104 | 73 |
| 2P8-RANTES | Q-G-P-L-S-G-Q-S-T-P | 660 | 96 | 71 |
| 2P9-RANTES | Q-G-P-P-G-G-Q-S-T-P | 7,800 | 82 | 62 |
| Third round of optimization | ||||
| Library 5 | Q-G-P-P-L-M-X-X-X-X | |||
| 5P1-RANTES | Q-G-P-P-L-M-W-L-Q-V | 18 | BD | BD |
| 5P2-RANTES | Q-G-P-P-L-M-W-L-Q-S | 29 | BD | BD |
| 5P3-RANTES | Q-G-P-P-L-M-W-M-Q-V | 18 | BD | BD |
| 5P4-RANTES | Q-G-P-P-L-M-W-M-Q-S | 27 | BD | BD |
| 5P5-RANTES | Q-G-P-P-L-M-W-T-Q-V | 21 | BD | BD |
| 5P6-RANTES | Q-G-P-P-L-M-W-T-Q-S | 19 | BD | BD |
| 5P7-RANTES | Q-G-P-P-L-M-A-L-Q-S | 17 | BD | BD |
| 5P8-RANTES | Q-G-P-P-L-M-S-T-Q-S | 26 | BD | 35 |
| 5P9-RANTES | Q-G-P-P-L-M-S-F-Q-S | 16 | BD | 12 |
| 5P10-RANTES | Q-G-P-P-L-M-W-L-Q-T | 21 | BD | BD |
| 5P11-RANTES | Q-G-P-P-L-M-W-R-G-S | 65 | BD | BD |
| 5P12-RANTES | Q-G-P-P-L-M-A-T-Q-S | 28 | BD | BD |
| 5P13-RANTES | Q-G-P-P-L-M-W-L-G-G | 34 | BD | BD |
| 5P14-RANTES | Q-G-P-P-L-M-S-L-Q-V | 26 | BD | 35 |
| 5P15-RANTES | Q-G-P-P-L-M-S-L-S-V | 34 | 43 | 50 |
| 5P16-RANTES | Q-G-P-P-L-M-G-L-S-V | 156 | BD | 18 |
| Fourth round of optimization | ||||
| Library 6 | Q-G-P-○-§-X-X-X-X-X | |||
| 6P1-RANTES | Q-G-P-P-G-G-G-G-L-G | 6,100 | 32 | 49 |
| 6P2-RANTES | Q-G-P-P-G-D-G-G-Q-V | 2,900 | 73 | 63 |
| 6P3-RANTES | Q-G-P-P-G-D-G-G-S-V | 280 | 85 | 66 |
| 6P4-RANTES | Q-G-P-P-G-D-I-V-L-A | 21 | 88 | 70 |
| 6P5-RANTES | Q-G-P-P-G-G-G-G-Q-S | 1,500 | 51 | 65 |
| 6P6-RANTES | Q-G-P-P-G-G-G-G-T-R | 3,000 | 52 | 69 |
| 6P7-RANTES | Q-G-P-P-G-S-W-S-S-V | 30 | 45 | 59 |
| 6P8-RANTES | Q-G-P-P-M-G-G-Q-V-T | 12,000 | 34 | 56 |
| 6P9-RANTES | Q-G-P-P-G-D-T-Y-Q-A | 10,000 | 41 | 55 |
| 6P10-RANTES | Q-G-P-P-G-D-T-V-L-W | 19 | 96 | 70 |
| 6P11-RANTES | Q-G-P-P-G-S-Y-D-Y-S | 79 | 90 | 69 |
| 6P12-RANTES | Q-G-P-P-L-G-A-G-S-S | 1,900 | 15 | 27 |
| 6P13-RANTES | Q-G-P-P-L-G-S-M-G-P | 390 | 23 | 36 |
| 6P14-RANTES | Q-G-P-P-L-D-F-G-G-A | 4,300 | BD | 35 |
| 6P15-RANTES | Q-G-P-P-M-G-G-T-S-A | 1,900 | 26 | 43 |
| 6P16-RANTES | Q-G-P-P-M-Q-G-G-L-S | 290 | BD | BD |
| 6P17-RANTES | Q-G-P-P-M-M-A-G-L-S | 29 | BD | BD |
| 6P18-RANTES | Q-G-P-P-L-Q-A-S-V-T | 1,900 | BD | BD |
| 6P19-RANTES | Q-G-P-P-M-S-G-H-S-T | 840 | 25 | 37 |
| 6P20-RANTES | Q-G-P-P-M-S-A-Y-Q-V | 160 | BD | BD |
N-terminal sequences of libraries and of isolated molecules are indicated. PSC, PSC-RANTES pharmacophore [n-nonanoyl-thioprolyl1-cyclohexyl2]; X, any amino acid; #, Ala, Pro, Ser, or Thr; ○, Gly, Leu, or Pro, [sct, Gly, Leu, or Met. Anti-HIV activity, CCR5 signaling activity, and CCR5 sequestration were screened using the assays described in Methods. BD, below the detection limit for the assay in question (see Methods). The reference compound is PSC-RANTES. Library 1, 1P1-RANTES, and 1P2-RANTES were first described in ref. 16.