| Primary Rat Microglia | Bz ATP Rat brain | AAV aSyn rat brain | Wt rat? | |
|---|---|---|---|---|
| P20: IC50 | 2.1 (0.8-6) nM, free Cave | |||
| P10: IC50 | 1.6 (0.4-7) nM, free Cav, (modelling) | |||
| P10: IC50 | 10 (4.1-24.3) nM, free Cav, (modelling, p25) | |||
| IL1b: IC50 | 7.6 nM | |||
| IL1b: IC90 | 68.4 nM | |||
| Cbr,u @4h - 3 mg/kg (MED) | 66 Nm (ie the last measurement after dosing) | |||
| Cbr,u @4h - 10 mg/kg (MED) | 65 nM (ie the last measurement after dosing) | |||
| Cbr,u AUC - 10 mg/kg (MED) | 639 nM*h | |||
| Cbr,u Cmax - 10 mg/kg (MED) | 198 ng/ml*H |
PK/PD
| Method | CNS 1 | CNS 2 | Peripheral | |||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
|
mice are pre-treated with an NLRP3 inhibitor (po) → BzATP (Infusion FC) → KILL & levels of IL-1β are measured in the FC , Dose PO TR06673219 — 200 µg BzATP Infusion FC — Necropsy, Measure IL-1 (FC section, ELISA) timeline: 0h → 1h → 24h | mice are pre-treated with an NLRP3 inhibition → 6ohda |
mice are pre-treated with an NLRP3 inhibitor (po) → LPS (0.1 mpk, ip, priming) → ATP (100 mpk, ip, for activation) → levels of IL-1β are measured in the plasma, Dose PO TR06673219 — 0.1mg/kg LPS, IP (priming step) — 100 mg/kg ATP, IP (activation step) — Measure (plasma, ?) timeline: 0h → 1h → 3h → 4h | ||||||||||||||||||||||||||||
| TR06673219 |
|
ID50 of 0.018 mg/kg, po [IC50 = 41 nM, Cp ...] (Figure 8).
|
PK/PD Strategy
- it will be important in having CSF measurements along with brain and serum that way we can build an understanding of the relationship between brain/CSF/serum levels in healthy vs disease. Since these ratios might be different in healthy vs disease.
- if we decide to use the IL-1b (serum) MDS-UPDRS correlation as our pathway modulation target. If we use this correlation, we will have to project the human serum measurements to CSF/brain levels to establish how much pathway modulation is required.
[Human target PK setting]
- 핵심 base for target setting: MCC-950, has demonstrated complete neuroprotection (TH counts similar to AAV empty vector mice) in the AAV/A53T mouse model when dosed 20mpk i.p. every other day (data not shown).
- RO(%) = -u.b / (K_d + C_u.b)
- FreeBrain_human = FreeBrain_mouse * (IC50_human / IC50_mouse)
| Compound (assay) | In vitro | Free Brain Target | |||
|---|---|---|---|---|---|
| IC50 (IL-1β suppression) | Ki | Cmax (nM) | AUC0-24 (nM h) | Max RO% | |
| MCC-950 (mouse primary microglia) | 17.8 (observed) | 12.65 nM (mouse Ki) | 90 (observed 겠지) | 300 | (=90/(...)) |
| TR06693098 (human microglia/neuron coculture, IPS) | 15.6 (observed) | 79 ← 90 × (15.6/17.8) (의미: MCC950 의 Cmax-mouse 90 은, 098 의 Cmax-human 에서는 79 이 상응하겠다. Target 이 됨) | 263 | (+90/12.65 × ...) | |
| TR06693098 (human primary microglia, PD) | 195 (observed) | 986 ← 90 × (195/17.8) (의미: MCC950 의 Cmax-mouse 90 은, 098 의 Cmax-human 에서는 986 이 상응하겠다. Target 이 됨) | 3,287 | 88 | |
| Cf) TR06693098 Mouse (primary microglia) | 11.3 (observed) | ||||
| Cf) TR06693098 (human primary microglia, PD) | 525 (observed) | ||||
| TR06693098 (Human) THP-1 cell IL-1b | 17 (oberved) | 43 (observed) | |||
MOUSE Free Brain Exposure
| Dose (mpk) | Cmax (nM) | AUC (nM/h) | Cavg (nM) |
|---|---|---|---|
| 30 | 882 | 3647 | 152 |
| 10 | 294 | 1216 | 51 |
| 3 | 88 | 365 | 15 |
| 1 | 29 | 122 | 5 |
HUMAN Free Brain Target
Correct for species potency differences. Use average across human assays
| Mouse Dose (mg/kg) | Cmax (nM) | AUC (nM/h) | Cavg (nM) |
|---|---|---|---|
| 30 | 824 | 3407 | 142 |
| 10 | 275 | 1136 | 48 |
| 3 | 82 | 341 | 14 |
| 1 | 27 | 114 | 5 |
Simulate various doses in GastroPlus to match human free brain Cmax, AUC, Cavg
50% Threshold in mouse brain PK/PD assay
(Inline Takeda PKPD slide: “C = Free PK/Potency (@1h)” IL-1β response (plasma) across compounds for TR06628513, TR06673219 (…), TR06647850, TR06673219 with regression annotated r² = 0.79, IC50 = 0.3 ± 0.2/0.4, e = 5.4 ± 0.3/0.6. Adjacent panel “Plasma PKPD Model · Predicted IL-1β response (brain)” shows TR06628513 and TR06673219 dose-response curves with model prediction shaded box. Bullets: “Prediction vs IL-1β reduction in the brain shows decent agreement for TR06628513 and TR06673219 despite different experimental setups”, “Brain IL-1β reduction was estimated based on plasma PKPD relationship with adjusted Imax (as observed in experimental studies)“. Slide preserved as body_r05 evidence.)
Postmortem
- Translational biomarker data
Proteina
[Proteina Original Study Proposal and Versions in progress]
|
Feasibility study, like Abbvie ($200k for feasibility study for a hundred sample), sample wanted. 100-10,000 cells , → 1ml (hundres ul) , rodent less, tens of ul, density relation to yoon taeyoung, Roche, who develops feasibility test protocol? I agree that ASC and GSDMD assay are advanced to the others. I'm also curious whether they can detected those two markers using human plasma and CSF collected from nigericin-treated rat for translational aspect. But I'm not sure their assay is compatible with rat ASC.. | |
|
Qs: pathogenic form 맞나? 교수? 더 확인없이 csf 가도 되나? How to 그 폼인지 확인 - cell system, & (particularly) buffer? -Co-IP? Antibody: purchase particularly for ASC, GSDMD To add: mouse, rat, brain/CSF/Plasma |
Uncertain Spans
| location | transcription | uncertainty |
|---|---|---|
| Top table p25 row | 10 (4.1-24.3) nM, free Cav, (modelling, p25) | The “p25” annotation could read “P25” or be an emphasis tag. |
| RO formula | RO(%) = -u.b / (K_d + C_u.b) | The numerator glyph appears as a faint “C-u.b” / “Cu·b” — leading negative sign is uncertain. |
| MCC950 Cmax cell | (90/(...)) | Closing fraction expression is partially cropped by the right edge of the cell. |
| TR06693098 IPS row | 90 × (15.6/17.8) | The numerator could read 15.6, 156, or 15·6 depending on decimal interpretation. |
| Plasma PKPD slide | r² = 0.79, IC50 = 0.3 ± 0.2/0.4 | The slide chart legend shows mixed glyphs; numbers above are best-effort reads from the slide thumbnail. |