| Rationale - disconnect: because in vitro and in vivo binding site conditions are different in terms of physiological substances near the binding site or feedback mechanisms that may affect binding. (Ting book) | |||
| 20230726 | ACUC committee, Imaging protocol, Rob (Takeda ex employee) FEPPA, & DPA714 both area ready to go at Dana Farber, TSPO longitudinal (~3w observation, to determine maximal uptake) → main study (202310) → result 202311 → PDE | Labelling imaging Led by TE Imaging | |
| 20230911 | (Parra, 2020 #2452) [NLRP3 PET] 6OHDA rat IACUC already approved, Surgery validated, already up & running Precursor to be synthesized for 11C labeling (old for 18F: stability x) (C not liked, but POC possible) (11c labeling is possible at Dana Farber) PET study in 202312, : surgery 20231211 → scanning 1(D1 dpa714, D3 FEPPA, D7 DPA714, D14 FEPPA, D21 DPA714) , Scanning 2 (reverse order) PE2I) → result 202401 (?), | [TSPO PET] AAV aSyn rat
| |
| 20240208 | 20240208: precursor has been shipped, [18F]DPA714 7 days post surgery confered the greatest signal window. | o [study1: time course] Ipsilateral AAV-aSyn and contralateral AAV-EV
-(satellite animal) | |
| 20240408 | The chemistry has been worked out that we have our collaborators at Dana Farber can actually make the C eleven tracer (NLRP3 PET 지칭) [plan] 6OHDA surgery at (Robbie Robertson) TBOS (or at Dana Farber) → transfer the animals to Dana Farber and they can actually do the image (C11 의 half life 짧아서 tbos 에서 못하고 imaging 는 Dana Farber 에서 해야함) |
| x days post surgery | x weeks post surgery | ||
|---|---|---|---|
| surgey | 4/15/2024 | ||
| Ship → TBOS arrival | 4/18/2024 | all 16) arrived safely recovery for 1 w | |
| 1st imaging | 4/23/2024 | scanning the first 5 rats today. 3 of which will be for the longitudinal assessment, the other 2 will be euthanized and brains harvested for IHC12 | 1.7 |
| 4/30/2024 | 19 | 2.7 | |
| 5/7/2024 | 26 | 3.7 | |
| 5/14/2024 | 33 | 4.7 | |
| 5/21/2024 | 40 | 5.7 | |
| 5/29/2024 | 47 | 6.7 | |
| 6/4/2024 | 54 | 7.7 | |
| 6/11/2024 | 61 | 8.7 | |
| 6/18/2024 | 68 | 9.7 | |
| 6/25/2024 | 75 | 10.7 | |
| 7/2/2024 | 82 | 11.7 | |
| 7/9/2024 | 89 | 12.7 |
[study 2: Tx impact]
rodent: dpa more metabolic activity (DPA with the DE fluoridation with the skull uptake, you can see, you can see that the the skull uptake here which is which is which is a result of DE fluoridation), FEPPA is better, human: vice versa
GOF mouse
| 20230111 | |||
| compound | [3H]TR06706493 | ||
| result | |||
| Next step | -Possible at-Risk in vivoPET experiment in 4-level 6-OHDA rats and NLRP3 GoFmice with in vivo candidates tracer ((for 18F-label).) -ARG study of patient samples using TR493 |
6OHDA Rat
| Initial (202202) | Optimized (202204) | Blocking study | |
|---|---|---|---|
| comp und | TR850 (iv); 0.01mg/kg at 1hr 이건 안 하는 듯 0.01mg/kg at 2hr (n=6) 0.1mg/kg at 4hr (n=6) 0.1mg/kg at 2hr (repeat from first study)(n=5) 다른 in vivo LCMS study 에서는 what dose? 많은 dose 쓰면 안 되나? | 6OHDA → (d10)blocker, Tool compound (TR06692993 would be the best compound as a blocker with enough exposure and good permeability & low clearance. Please share estimated brain free conc of TR993 at 5 PO in rats): PO 30mg/kg → after 1hr(Cmax), tracer (TR06647850 IV) → 4h after, tissue collection | |
| meli | surgeries beginning of April, → dosing will be mid-April. Samples will be collected on 13th Apr (TSD time) → The concentrations of TR850 will be generated next week | 1 week habituation , → the dosing will be on 25th, July → the samples (plasma and striatum) will be obtained on 1st , August → PK data will be available around the middle of August. @Rolzin, Paul May I ask to measure the NLRP3 protein level in the striatum again?, because this is the first time study at CRO and we would like to confirm the reproducibility of NLRP3 increase in the LPS model. | |
| The brain surgery might be affected the BBB penetration. As control, sham surgery would be desirable? After confirming an appropriate regimen of TR850, team will discuss on the protocol for the blocking study of TR850 with CN candidate. | Fold change is similar between vehicle treated and '993 treated animals → significant blocking not observed. The increase in TR850 concentration at the injection site may be due to the disruption of morphological change of the BBB barrier caused by 6OHDA injection. ✓ 29 Jun 2022 2022 年 6 月 29 日 14:11 Untitled Section Page 1 change of the BBB barrier caused by 6OHDA injection | ||
| ult | (small NLRP3 WB bar-chart inset, kept as evidence) NLRP3 (WB: ↑ 2x at ?mg, x h?) | ||
| -Maybe ↑ SB -Paul: I do worry about changes in nonspecific binding (eg. any tissue morphology changes in the striatum and that what we are seeing is not specific binding (due to increased NLP3 expression). | Blocking study plan: |
LPS (systemic) Rat
| 1st | 2nd | 3rd | |
|---|---|---|---|
| comp | Tracer (850) dose: (0.1 & 1 mpk LPS dose: (2 mpk tissue collection time: 1& 2 h | ↓ Tracer (850) dose: (0.1 & 1 mpk → ) 0.01 & 0.1 mpk ↓ LPS dose: (2 mpk → ) 0.5 mpk ↓ tissue collection time: 1& 2 h ) → 2 & 4 h | LPS (ip): 0.5 mpk, 4 days → IV (different tracer) TR06683014 and TR06683015 (iv) , → tissue collection after 1,2h |
| li | The brain samples in this study were shipped to TSD Paul to check the NLRP3 expression levels. The results of NLRP3 expression will be generated next week. | ||
| 3 | Plasma and striatum concentrations of TR06683014 and TR06683015 at 1 & 2 hr of 0.01 mpk in LPS rats was generally comparable with ~10ul... | ||
| ult | ↑ NLRP3, ↑ IL-1b, =TH | TR06647850: Fold increase at 4 hr of 0.1 mg/kg in LPS rats was observed as well as 6-OHDA rats. However, there was no difference between control LPS rats but no increase was observed in < mg/kg LPS rats. The results of the 2 mg/kg group was reasonable because the final dosing of LPS was | There was no big difference of Kp... |
Uncertain Spans
| location | transcription | uncertainty |
|---|---|---|
| 1st imaging schedule, “x weeks post surgey” column | initial value 1.7 next to 4/23/2024 | The 1.7 weeks figure is calculated from 12 days post-surgery; preserved as-seen. |
| 6OHDA Rat ult row | inline NLRP3 WB bar-chart | Bar chart with multiple bars (cell numbers not legible) is kept as evidence; the caption “NLRP3 (WB: ↑ 2x at ?mg, x h?)” is preserved. |
| LPS (systemic) Rat 3rd column tail | ”There was no big difference of Kp…” | Tail of the cell continues beyond the photo; the rest is cropped at the page edge. |
| 20240208 row | ”[18F]DPA714 7 days post surgery confered the greatest signal window." | "confered” preserved as misspelled in original. |
| Bottom-left “comp und / meli / ult / li / 3” row labels | leftmost row identifiers | Truncated in the photo; preserved as-seen. |