| Dog | Dog Acute Escalating Dose Toxicity and Toxicokinetic Study | Rat pk data for 30, 100, 300mpk 는 있는데 |
| Dog | 2w DRF Study | |
| Dog | Dosage Selection: Dog CV | |
| Dog CV | Safety Pharmacology Study, single dose? |
| Dose Group (mg/kg/d) | D8 or D14 Tmax (h) | D8 or D14 AUC0-24 Total (ng*h/mL) | Exposure Margin (D8 or D14 AUC0-24 / AUCeff) |
|---|---|---|---|
| 0 (Vehicle) | NA | NA | NA |
| 30 | 1 (M)/1 (F) | 81,400 (M)/117,000 (F) | 0.94 (M)/1.35 (F) |
| 100 | 3 (M)/3 (F) | 184,000 (M)/306,000 (F) | 2.12 (M)/3.52 (F) |
| 600 terminated early after Dose 8 | 1 (M)/1 (F) | 913,000 (M)/1,080,000 (F) | 10.5 (M)/12.4 (F) |
AUCeff is the human efficacious exposure (233,333 nMh or 86,902 ng*h/mL) predicted using in vitro potency data in primary microglia and in vivo exposure in AAV1/2-hA53T-aSyn mice at the efficacious dose of 30 mg/kg.
Based on the current tox and efficacy profiles of TR06698660, we do not have sufficient exposure margins to proceed the compound towards the candidate selection.
| Rat 2w tox | Atuka 1st | Atuka 2nd | |
|---|---|---|---|
| 600 | |||
| 100 | |||
| 30 | 30 | 30 | |
| 10 | exp margin | ||
| MTD | 100 mpk | real clean? (lung, PL) 184,000 from Atuka1st 86,902 2.12 |
Accumulation Ratio
R = Day21 AUC0-24 / Day 1 AUC0-24
| Day21 AUC0-24 | Day1 AUC0-24 | R | |
|---|---|---|---|
| TR06693098 | |||
| 10 mg/kg | 34800 | 21500 | 1.62 |
| TR06698660 | |||
| 10 mg/kg | 12800 | 11300 | 1.13 |
| 30 mg/kg | 28900 | 29900 | 0.97 |
| Ceff | Min E D from Atuka | |
|---|---|---|
| 30 mpk | ? | |
| 10 mpk | neuroprotection will be 70/3 then only 23% | ~6? |
| future in AAV rat, | ? | |
| 3 mpk | neuroprotection will be 70/10 then only 7% | ~20? |
Ames test
CVB: Reactive metabolites
Good review; (Thompson, 2016 #2207)
Heather presentation 202209 15 core (Takakusa, 2008 #2141)
| Phenol structure 가 잘 형성 | Reactive metabolite | RMs bind covalently to proteins and DNA, function (LIVER), Toxicity (frequently idiosyncratic AE) | |
|---|---|---|---|
| Background: | (Chemical trapping agents, such as reduced glutathione (GSH), can form stable adducts with many reactive species. Trapping agents, incubated with liver microsomes, are routinely used in the identification of RM.) → Following its formation, the trapped reactive metabolite can be analyzed via various analytical approaches most commonly with LC MS or fluorescence or radiochemical detection | ||
| Dose |
<100 mg/day → if exceed, ratios (ORs) of 7 then CVB assay is needed were found for daily doses ≥ 100 mg CVB burden (=Daily body burden for reactive metabolites = Drm, in {Thompson 2016}) CVB buden = fcvb (fraction of RM formation) x Daily Dose Example of TR098: fcvb was 0.0848 Drm = D · fa · fm · frm D is the total daily dose (mg/day), fa is the fraction of the dose absorbed, fm is the fraction of the dose eliminated via metabolism, and frm is the fraction of the metabolism leading to thiol-RM. CVB burden (dose 아니라) <1 mg/day (CVB assay 필요인가 봄) ({Thompson 2016: with a cutoff of 1 mg, the overall sensitivity and selectivity for distinguishing the hepatotoxins from the nonhepatotoxins would be 67 and 71%, respectively) 1 mg/day = 0.0848 x Dose, so Dose has < 0.0848/1, 즉 < 11.8 mg (= allowed humen dose), | ||
| CVB level |
liver microsomal incubations using the radiolabeled drug) <50 pmol/mg protein covalent binding | ||
| finding | Low risk based on RI- and MS- GSH Met-ID in HLM. Risk with 2 trace adducts in rat S9 MS GSH | ||
| plan |
-CVB (covalent binding) studies of 14C labeled compound (results 20230501, DSRE recommendation: do this before CS!) -pending HED -Assess liver weight and IHC in Atuka mouse study (9 wks dosing 3, 30 mg/kg) • 14 day DRFs and 28 day GLP studies | ||
CVB Activities
Milestone:
- Initial HED prediction
- CVB Pre-cursor (5g)
- 14-C labeled compound
- Run CVB assay
- CVB Burden Threshold Determination (DMPK)
- Approx start to first GLP in vivo study
Based on current timeline:
- Expect 14C-CVB data by May 1, 2023
- Need HED to calculate CVB Burden
- Need to have CVB Burden threshold also defined to interpret risk
- GLP study starts expected beginning May based on API timing (scheduling pending)
- Clinical assessments: Standard clin path
| < 2 positive hits in aggregated in vitro panel score | Scenario | DSRE Recommendation |
|---|---|---|
| If the effective dose in AAV-A53T-αSyn efficacy study is 30 mg/kg | Estimated HED is 10 mg/day | Keep CS: CVB data is not required at CS |
| Estimated HED is 10-100 mg/day AND phospholipidosis seen in the Atuka study | Postpone CS: CVB data is required at CS* (ie CS delay 결과가 may 에 나오니까) | |
| Estimated HED is >100 mg/day |
AUCeff: AUC(0-24) in the AAV1/2-hA53ThA53T-aSyn mice administered 30 mg/kg/day TR06693098 for 21 days, (76,100 ng*h/mL)
Ocular
al carotid a → ciliary body → aques humor → cornea
Phospholipidosis
Drug induced phospholipidosis (DIPL) is a phospholipid storage disorder characterized by an abnormal accumulation of multi-lamellar bodies (myeloid bodies) in cells and tissues
Myeloid bodies can be seen in thin tissue sections, peripheral blood cells, and urinary sediment using transmission electron microscopy (TEM). They are characterized by concentric layers of electron dense, membranous material (whorls) surrounded by a single limiting membrane. Myeloid bodies occur naturally in the late endosomes/lysosomes (LE/Lys) of some tissues where they act as storage vesicles for secreted and undigested lipids and protein myeloid bodies also serve as repositories for excess drug/drug metabolites and undigested drug-phospholipid complexes
- TR06698660 → possibility of phospholipidosis (PLsis)-related findings in female after 5-days at 600 mg/kg, where Cmax is 63,200 ng/mL and AUC0-24 is 1,300,000 ng*h/mL.
- TR06693098 → possibility of Plsis-related findings in male after 3-days at ≥100 mg/kg, where Cmax is 66,914 ng/mL and AUC0-24 is 1,238,167 ng*h/mL.
- TR066980443 → No indications of PLsis-related findings in male after 3-days up to 600 mg/kg, where Cmax is 119,000 ng/mL and AUC0-24 is 3,130,000 ng*h/mL.
- ü In only one rat at 600 mg/kg, decrease in spontaneous movement and/or ataxia was found and the reason is unclear. Longer duration study (14-d tox) is needed to investigate this profile. Spontaneous movement and/or ataxia would be non-specific toxicity. This finding is visible due to TR098 dosing, and a certain number of similar cases will be specified for each cohort of animals in its 14-d tox study.
Uncertain Spans
| location | transcription | uncertainty |
|---|---|---|
| Phospholipidosis bullet | TR066980443 | Compound id TR066980443 differs from the document’s other 8-digit compound ids (e.g. TR06698660 / TR06693098); the visible text shows the longer 9-digit string and is preserved as visible. |
| dose escalation | 913,000 (M)/1,080,000 | The first row of the dog dose-group table has 600 mg/kg 913,000 (M)/1,080,000 (F) but the AUC value alignment is partly obscured by a column boundary; preserved as visible. |