<sup>* A survey of the US FDA, Canadian, UK MHRA, and European HMA drug Labels, monographs, and summary reviews.
* DIPL suspected but not confirmed by TEM.</sup>

• di-22:6-BMP
  • background
    • Bis(monoacylglycero)phosphate (BMP) is a negatively charged glycerophospholipid with an unusual sn-1’ structural configuration.
    • BMP is primarily enriched in endosomes/lysosomal membranes.
    • BMP is thought to play a role in glycosphingolipid degradation and cholesterol transport.
    • Elevated BMP levels are found in many LADs, suggesting an association with lysosomal storage material.
  • Rationale
    • Animal {Thompson, 2012 #2183, Liu, 2014 #2151}: d-22:6-BMP urine in animal samples from animals treated with drugs that induce TEM-confirmed PLD-13 (statistically d-22-6-BMP in urine, {Liu, 2014 #2151})
    • Rat: {Thompson, 2012 #2183, 1 rat, DIL422} → ↑ tissue phospholipidosis (IHC) at 4 di-22-6-BMP in urine ↑ 자전 specific 한다 (nephorotox supplementary 에서 reported).
  • Normal range
    • Human: {Liu, 2014 #2151) NPC
    • {Liu, 2014 #2151} urine in HC, plasma & serum & urine in human, rat, mouse, dog, monkey
    • {Akgoc, 2015 #2192} MW of d-22-6-BMP corresponds to 865.5

	{Liu, 2014 #2151}	{Akgoc, 2015 #2192}
CSF	HC: 87.0 ± 130.5 PD: 380.5 ± 174.6 nM CO (Tomas) HC: 14.36 ± 15.94 ng/ml DIL: 19.85 ± 13.49 nM
Plasma	0.29 ± 0.15 ng/ml (range: 0.11-0.79 ng/ml) (MW 865.5 도) HC: 39.66 ± 22.43 nM PD: 19.65 ± 9.34 nM
serum	0.46 ± 0.52 ng/ml (range: 0.07-2.7 ng/ml)
urine	creatinine-adjusted concentration: HC (n=20): 0.86 ± 1.13 ng/(mg creatinine)) vs NPC (n=21): 44.3 ± 60.9 ng/(mg creatinine).

• Points
  • Normal & (normal 이 data미디 HC vs NPC is clear),
  • Normal range 를 sato-san data 에서 봅고 ◇ 25D 를 predictors

→ i) animal: elevated cases 가 tissue DIPL 와 match 되는지.
→ ii) clinical monitoring plan: abnormal range 를 보이면 monitor (blood cell TEM, long-term monitoring, liver/renal function testing)

• PD therapeutic window:

  • DIPL is tissue-specific
  • PD patients have ↑ p… PD, ↑BMP?

• [20221208] The DSRE explored the utility of di-22:6-BMP as a potential biomarker for drug-induced phospholipidosis.

  • Di-22:6-BMP is a promising, exploratory biomarker for phospholipidosis but is not validated at this point. To determine its utility in our program as nonclinical and clinical biomarker, further works may need needed.
    • The biomarkers should be evaluated in nonclinical repeat-dose toxicity studies (≥ 2-week duration) to assess the correlation of this biomarker to our NLRP3 compound-induced phospholipidosis.
      • Detailed methods are not available from the published literature and methods development work is likely needed.

• Clinically, transmission electron microscope assessment of tissue biopsy or blood smear samples can be used to identify the phospholipidosis - may be challenging to implement though

• There are other programs working on lysosomal dysfunction where other PD biomarkers were explored. Consider exploring how these respond in the safety studies - extra blood samples will be collected from the rat for the 14-day for this purpose

  • Glucosylsphingosine - previously used as PD biomarker
  • Cathepsin D - TSHO has validated the assay for this biomarker
  • LAMP1 (as a fluid biomarker)
  • Di-22-6-BMP - May be worthwhile exploring this internally

• Lipofuscin detection in eyes has also been used in the past for detecting the phospholipidosis. However, if we see such findings in eyes/brains, the progression of the compound would be challenging

• Action item:

  • Top biomarker candidates will be explored in the exploratory blood samples collected from the rat 14-day study.
  • Arthur/clinical team will review and discuss the summary table that Heather put together on the use of CAD-containing drugs in PD patients and share their inputs from the clinical perspective.

Established NLRP3 Inhibitor Screening Flow

(Multi-coloured flowchart figure - kept as evidence in body_r03_c01.jpg, body_r04_c01.jpg, body_r04_c02.jpg. Visible node labels reproduced in format_notes. Footer: Takeda Pharmaceutical Company Limited - Confidential, for internal use only.)

ShareFolder

TCAL/Departments/CNS/PROJECTS folder

NLRP3 Inflammasome > QS M&S > Slides-Hamid

☐ NLRP3 Inflammasome

InterACT-RAD-Pipeline / NLRP3 Inflammasome / NLRP3 CN Candidate folder

NLRP3 CN Candidate Folder ATUKA Vol-1

2023

☐ NLRP3 Timeline toward CS_202308.pptx

☐ Timeline NLRP3 CN Candidates.xlsx

☐ NLRP3 CNCS candidate assay list.xlsx

Uncertain Spans

location	transcription	uncertainty
Rationale / Rat citation	reads Rat: {Thompson, 2012 #2183, 1 rat, DIL422}; the DIL422 token may be a study/dose code; preserved verbatim.	low confidence on DIL422 token.
Normal range CSF / Akgoc nM values	the CSF and Plasma rows mix multiple literature sources (Liu 2014 #2151, Akgoc 2015 #2192) and both unit conventions (ng/ml and nM); preserved verbatim.	low confidence on per-source row alignment.
Established NLRP3 Inhibitor Screening Flow / node labels	the screening flow diagram has many shaded nodes with small labels (Cyp Inhibition / Promiscuity / Na channel / Solubility / Mouse Plasma Protein Binding / Rodent brain homogenate Binding / Mouse Hepatocytes and Build IVIVC / Safety47 / Proarrhythmic assay / Liver tox assay (indpnero) / Genotox (Bluescreen)); per-node arrow direction and IC50 thresholds are reproduced in format_notes with low confidence on the exact arrow topology.	low confidence on chart topology and arrow direction.
ShareFolder / SharePoint URL fragments	the SharePoint URLs are partly clipped at the right edge of the cell and contain query parameters (csf=1&e=...&cid=...&FolderCTID=...&View=...); the visible portions are reproduced as anchor text and may not be reachable as-is.	URL fragments clipped; partial.