text stringlengths 1 5.46k |
|---|
Project Activities, Timelines and Deliverables by Stage |
Research Program |
The scope of the Research Program includes activities from the exploratory discovery phase (Stage 1 of the Research Program) to the candidate nomination phase prior to initiation of GLP toxicology (Stage 2 of the Research Program). The goal of this research is to validate DUB targets for their role in Tau clearance and... |
Stage 1: DUB In Vitro and In Vivo Target Validation. Mission Library Screening and Limited Chemistry |
Overview: Stage 1 of the Research Program includes activities toward the in vitro and in vivo validation of DUB targets for Tau and /or α-synuclein clearance, as well as the screening of Mission's Compound Library and the generation of potential tractable chemical series for up to four (4) DUBs. The activities within S... |
[Note: There appears to be a diagram in the original document showing the flow from "In Vitro Target Screening & Validation" to "Pre-Selected DUBs*", "Initially Selected DUBs", and "Mission Library Screening & Tractable Chemical Series and In Vivo Target Validation"] |
Mission Activities:<br>• Pharmacologic phenotypic screen for -syn clearance (SHSY5Y, REN Cell VM and iPSC-derived Dopa neurons). Including confirmation of target selectivity by siRNA knock down (KD) experiments.<br>• Genetic CRISPR phenotypic screen for -syn clearance (SHSY5Y), including sequencing confirmation to asse... |
*Initially Excluded DUBs comprise USP30, UCHL1, Cezanne, USP28, USP4, USP10, USP35, USP34, TRABID, USP1 and USP21 |
AbbVie Activities:<br>• Pharmacologic phenotypic screen for Tau clearance (SHSY5Y and iPSC-derived neurons). Including confirmation of target selectivity by siRNA knock down (KD) experiments.<br>• Genetic CRISPR phenotypic screen for Tau clearance (SHSY5Y)<br>• Confirmatory siRNA on iPSC-derived neurons on selected DUB... |
Stage 1: Activities and Timeline Assumptions |
Stage 1A Activities |
Stage 1A activities are initiated with two types of DUB phenotypic screens for misfolded protein clearance: in vitro genetic CRISPR screening and in vitro pharmacologic screening. These in vitro screens will occur in parallel for Tau clearance and for α-synuclein clearance. During Stage 1A, Mission will be responsible ... |
Mission will conduct the in vitro α-synuclein target validation studies (i.e. genetic CRISPR and pharmacologic phenotypic screens) which will be conducted as described below. CRISPR studies will be undertaken in SHSY5Y cells, the pharmacologic phenotypic screen will be undertaken in SHSY5Y, REN Cell VM and iPSC-derived... |
• 72 potent (10-300 nM) inhibitors of Pre-Selected DUBs with generally >10x selectivity over the DUBs tested by Mission. These include inhibitors of 19 DUBs (USP1/6/7/10/13/14/21/25/28/30/46/47; UCHL1/3; BAP1; SENP1/2/6 and Cezanne) spread throughout the USP, UCH, OTU and SENP sub-families.<br>• 58 less potent (300 nM ... |
For the pharmacologic phenotypic screen, Mission will conduct a confirmatory target selectivity siRNA knock down (KD) experiment to check the effect of a compound on α-syn in the absence of a given DUB. Ideally, if the compound effect is specific to a given DUB, then its effect will be significantly affected when the D... |
Mission will conduct a confirmatory iPS cell siRNA screen on selected DUBs. Initial screen will be a confirmatory screen on the DUBs that are positive from the in vitro genetic CRISPR screen and the in vitro pharmacologic phenotypic screen. In addition, iPS cell siRNA screening will be conducted on DUBs for which knock... |
In addition to the activities described above, Mission will provide AbbVie with pre-aliquoted plates with ~ 240 compound set to enable the pharmacologic phenotypic screen with Tau clearance read–out by AbbVie. |
During Stage 1A of the Research Program, AbbVie will be responsible for the in vitro Tau target validation studies (i.e. genetic CRISPR and pharmacologic phenotypic screens) with Tau clearance assay. The in vitro genetic CRISPR phenotypic screen will be conducted using the SHSY5Y cells (expressing P301L mutant tau and ... |
AbbVie will conduct a confirmatory iPS cell siRNA screen on selected DUBs. Initial screen will be on the DUBs that are positive from the in vitro genetic CRISPR screen and the in vitro pharmacologic phenotypic screen. In addition, iPS cell siRNA screening will be conducted on DUBs for which knockout resulted in cell le... |
The duration of the activities associated with Stage 1A is ~ 9 months including assay development and completion of the DUBome phenotypic screens. All in vitro target validation work will be conducted in parallel by the responsible teams. Upon completion of these in vitro studies, up to eight (8) DUBs will be selected ... |
Without limiting AbbVie's decision-making rights, the following guidelines will be considered for the ranking of DUBs based on data generated from: (1) the in vitro pharmacologic phenotypic screens, (2) the in vitro genetic CRISPR phenotypic screens and (3) the confirmatory iPSC cell siRNA screens on selected DUBs (DUB... |
DUB Selection Guideline for Stage 1A selection of up to eight Initially Selected DUBs |
1. Pharmacologic Phenotypic Screen<br>a. At least 50% decrease of Tau or α-synuclein (statistically significant) at a "selective" concentration (>5-fold) in replicated assays (at a minimum n>2)<br>b. Separation of Tau / α-syn clearance and cytotoxicity (toxicity ~15% or less)<br>c. Demonstrated dose response (IC50)<br>... |
2. Genetic CRISPR Phenotypic Screen<br>a. Statistically significant effects regarding both the decrease of Tau/α-synuclein and target KD in replicated assays (at a minimum n >2)<br>b. > 70% knock down by PCR and sequencing / western blot, if point (a) is met<br>c. Greater than 50% decrease of Tau / α -synuclein in repl... |
Other points for consideration include: (1) supportive data in confirmatory iPSC screen (pharmacologic and / or genetic), (2) additional compounds showing decrease of Tau / α-synuclein and (3) literature support. |
Stage 1B Activities |
The activities of Stage 1B are focused on the screening of Mission's Compound Library (~ 12,000 compounds), limited chemistry expansion efforts to assess the chemical tractability of hits, assay development and the in vivo validation studies for Tau and / or α-synuclein. |
During Stage 1B, Mission will be responsible for Mission's Compound Library screening, limited hit expansion chemistry and development of all primary biochemical assays (e.g. fluorescence intensity, fluorescence polarization or other technologies allowing the assessment of DUB activity) and / or appropriate cellular as... |
Mission will screen the Mission Compound Library for compounds that Bind to the Initially Selected DUBs. This screening will be done as SPS at 10 and 100 μM. Determination of IC50 will be done on any hits with > 40% and > 80% inhibition to the test concentration, respectively prior to subsequent hit expansion. Mission'... |
In the situation, in which Mission's Compound Library has already been screened for any of the Initially Selected DUBs, Mission will screen any additional compound sets that were not included in that first screen for such target (e.g. newly synthesized compounds, purchased and / or acquired compounds, etc.). The estima... |
For those Initially Selected DUBs for which biochemical and / or cellular assays are not available, Mission will develop these assays during Stage 1B. In the event an antibody is not validated or is not commercially available to be used in the development of the cellular assay, Mission will conduct the following activi... |
Therefore, there are three potential Mission chemistry timeline scenarios based on the availability of reagents and / or assays as indicated in the table below:<br>[Table showing: Scenario 1: Reagent Status - Available, Assay Status - Developed, Timeline - ≤3 months]<br>[Table showing: Scenario 2: Reagent Status - Avai... |
Estimated timeline assumptions: Generation of reagent ~ 3 months, assay development ~ 2 months and assay transfer to AbbVie ~ 1 month. |
This screen will be performed multiple times during the course of Stage 1B but at a minimum, selectivity in Agreed Assays (refer to Schedule 1.15) and potency against Initially Selected DUBs will be determined to identify compounds that Bind to Initially Selected DUBs informing on which compounds and DUBs to pursue for... |
Stage 1B Minimum Deliverables for Mission as described in Schedule A to this Schedule 1.157 below. |
In parallel to the Mission activities, AbbVie will initiate the in vivo target validation using shRNA-AAV knock down. The in vivo target validation will be done in a staged manner (i.e. 4 DUBs at a time). Studies can be done by AbbVie or at a CRO and are estimated to take ~ 6 months. |
AbbVie will review the Stage 1 data (Stage 1A and Stage 1B) to decide on the 4 Selected DUBs for transition into Stage 2. At AbbVie's discretion and upon paying the Stage 1 Fee for such DUB, Selected DUBs may be moved to Stage 2 in a rolling over process at such time that data generated supports moving into the next St... |
Stage 1B DUB Selection Guideline: |
• In vitro target validation completed per the Stage 1A Guideline Criteria<br>• In vivo target validation completed on Stage 1B<br> o At least 50% knock-down of target in relevant cell-type (qPCR or RNAscope)<br> o Significant reduction of Tau / α-syn pathology in transgenic (Tg) animal model – shRNA (~30% or more, sta... |
Data for each DUB target will be shared as it becomes available to facilitate decision-making. |
Stage 1: Summary of Activities |
[Table with columns "Mission" and "AbbVie"]<br>Stage 1A<br>Mission:<br>• Pharmacologic phenotypic screen for α-syn clearance (SHSY5Y, REN Cell VM and iPSC derived Dopa neurons). Including confirmation of target selectivity by siRNA knock down (KD) experiments<br>• Genetic CRISPR phenotypic screen for α-syn clearance (S... |
Stage 1B<br>Mission:<br>• Biochemical, cell-based, selectivity assay and reagent development (as needed).<br>• Biochemical screening of Mission's Compound Library and limited chemistry expansion on up to 8 DUBs<br>• Biochemical assays will be transferred on a DUB by DUB basis once a Selected DUB is chosen to initiate S... |
Stage 1 Deliverables: Refer to Schedule A to this Schedule 1.157 below for Minimum Deliverables for Stage 1A and Stage 1B |
Stage 1: Representative Timelines for Each Scenario<br>[Note: The document appears to contain timeline graphics/charts for Scenarios 1, 2, and 3 that I cannot reproduce in text format] |
Stage 2 |
Overview |
Stage 2 is designed to generate the discovery and preclinical data required for candidate Licensed Compounds to be ready to enter GLP toxicology studies. Activities will be initiated with the screening of all or a portion of AbbVie's Compound Library and medicinal chemistry efforts (HTL and LO) on selected compounds th... |
[Note: The document appears to contain a diagram showing the flow from "Up to 4 DUBs" through "ABBV Library Screen & Hit to Lead" and "Lead Optimization" to "Candidate Nomination"] |
Mission Activities<br>• Participation in CLOOC<br>• Assay transfer to AbbVie<br>• Primary biochemical assay, selectivity assays in Agreed Assays (Schedule 1.15) and cell-based target engagement assays, in vitro efficacy assays (Tau and α-syn)<br>• Ex vivo target engagement assays<br>• Full DUB selectivity screen as req... |
AbbVie Activities<br>• Participation in CLOOC<br>• AbbVie's Compound Library screen<br>• HTL and LO chemistry on Mission and AbbVie Hits<br>• Compound synthesis<br>• In vivo efficacy in transgenic (Tg) mice (PoC) (Tau and -syn clearance)<br>• Brain penetration, PK, ADME & safety<br>• Advanced safety characterization (N... |
Stage 2: Medicinal Chemistry Activities and Timeline Assumptions |
Prior to the initiation of AbbVie's Compound Library screening and Medicinal Chemistry activities, Mission will have transferred to AbbVie the necessary in vitro assays (e.g. biochemical assay) specific to the Selected DUBs (up to 4 DUBs) from Stage 1. These assays for the up to 4 Selected DUBs should be developed (if ... |
AbbVie will have responsibility for AbbVie's Compound Library screening and for the medicinal chemistry efforts toward Lead Generation (LG) and Lead Optimization (LO) of those compounds that Bind to a Selected DUB. Also, AbbVie will be responsible for the medicinal chemistry activities toward Lead Generation (LG) and L... |
Mission will perform the following in vitro activities: primary biochemical assays, selectivity assays using Agreed Assays for determining any compound (un)restrictions as well as larger DUB screen for determining full selectivity, cell-based target engagement assays with activity probes and in vitro efficacy assays (i... |
Mission will conduct kinetic analysis to determine the kinetic behavior of leading representatives of each chemical series by measuring Kon and Koff. This information will help with the generation of differentiated chemical series in the early stages of the medicinal chemistry optimization activities. |
During Stage 2, Mission shall provide all research results, research data and written reports relating to to the activities carried out by Mission during Stage 2, including all results of any screens of Mission's Compound Library to determine any Mission Compounds that Bind to the Selected DUBs and any DUB Selectivity ... |
Stage 2 activities including chemistry activities will be under the strategy, set by the "Collaboration Lead Optimization Oversight Committee" (CLOOC). |
Within Stage 2, AbbVie and Mission will be mutually responsible for target validation and potential mechanism of action (MOA) characterization studies as soon as the Selected DUB targets are known and assays and tools are available. |
Within Stage 2, AbbVie and Mission will jointly be responsible for the development of translational pharmacodynamic endpoint biomarker assay(s) as soon as the Selected DUB targets are known. Taking into consideration each party's areas of expertise, Mission will lead the efforts toward the preclinical development activ... |
A desirable target engagement (TE) biomarker assay(s) should meet the following characteristics:<br>• Provides evidence of central exposure and binding at target (desirable but not required)<br>• Proves that binding results in a dose-dependent mechanistic activity. For example for a DUBi – ubiquitination status of a re... |
The MOA characterization, target validation and target engagement biomarker assay development activities are DUB specific and will be defined in a case-by-case basis during Stage 2. |
Stage 2 Mission and AbbVie Activity Overview – Actual screening funnel is highly dependent on the Selected DUB<br>[Note: The document appears to contain a flowchart/diagram showing the screening and selection process with various steps including Primary Biochemical Assay, Tier 1 In vitro ADME, Cell-based target engagem... |
Stage 2: Summary of Activities |
Mission:<br>• Participation in CLOOC<br>• Primary, selectivity assays in Agreed Assays (Schedule 1.15) and cell-based target engagement assays, in vitro efficacy assays (Tau and α-syn)<br>• Ex vivo target engagement assays<br>• Agreed Assays as needed<br>• Full DUB selectivity screen as requested<br>• Target validation... |
AbbVie:<br>• Participation in CLOOC<br>• AbbVie's Compound Library screen<br>• HTL and LO chemistry on Mission and AbbVie Hits<br>• Compound synthesis<br>• In vivo efficacy in transgenic (Tg) mice (PoC) (Tau and α-syn clearance)<br>• Brain penetration, PK, ADME & safety<br>• Advanced safety characterization (Non-GLP) –... |
Stage 2 Minimum Deliverables: Refer to Schedule B to this Schedule 1.157 below. |
Stage 2: Representative Timeline based on Scenario 1 from Stage 1<br>[Note: The document appears to contain a timeline chart/diagram] |
Schedule A to Schedule 1.157 |
Stage 1A Minimum Deliverables and Stage 1B Minimum Deliverables |
Stage 1A Minimum Deliverables |
For the Stage 1A activities Mission will deliver the following information and report(s) of the generated data to AbbVie. |
Stage 1A: Protocol(s), reagent(s) and data deliverables |
1. Pharmacologic Phenotypic Screen (SHSY5Y, REN Cell VM and iPSC-derived Dopa neurons)<br><br>a. For each cell line, Mission will provide a file (Excel,Prism) containing the SPS screening data with the percent α-synuclein clearance and percent cytotoxicity for each compound and at each concentration tested. Data from r... |
2. Genetic CRISPR Cas9 Phenotypic Screen (SHSY5Y)<br><br>a. A file (Excel, Prism) will be provided containing the percent α-synuclein clearance and percent viability (nuclei count) for each DUB knockout (Pre-Selected DUBs as per Schedule 1.141) relative to the negative control (non-targeting or scrambled gRNA). Data fr... |
3. iPS cell siRNA screen on a selected DUB basis<br>a. A file (Excel, Prism) will be provided containing the percent α-synuclein clearance and percent viability (nuclei count) for each DUB identified as positive in a small molecule (three cell lines) assay as per above 1b and/or as positive in the CRISPR SHSY5Y genetic... |
4. Provide AbbVie with pre-aliquoted plates containing the ~240 compound set<br><br>a. Mission will provide 50-100 µl of 10 mM DMSO stock of the ~ 240 compounds in pre-aliquoted plates (1.4 mL 2D barcoded matrix tubes with push caps). In addition, an excel file containing the plate number (1-3), well position (A1-H12),... |
5. Provide AbbVie with protocols and study / experimental designs and SOPs. |
For the Stage 1B activities Mission will deliver the following information and report(s) of the generated data: |
Stage 1B: Minimum Deliverables |
1. Biochemical screening of Mission's Compound Library (approx. 12,000 compounds) against up to 8 DUBs selected from Stage 1A.<br><br>a. Scenario 1 (refer to Stage 1B Activities of Initial Research Plan, Schedule 1.157). A file (Excel) will be provided containing the % inhibition at 10 and 100 µM as well as 8-point hal... |
2. Cellular Ubiquitin based activity probe assays (SHSY5Y or other relevant cell line).<br><br>a. Data will be provided in a report (Powerpoint) on the validation of commercially available antibodies or existing Mission antibodies against the relevant DUB in cells following knockdown and/or knockout of the respective I... |
3. Agreed Assays for Initially Selected DUBs.<br><br>a. A file (Excel) will be provided containing the % inhibition at 10 and 100 µM and IC50s for up to ten (10) active compounds per Initially Selected DUB, if requested for the DUBs as outlined in Schedule 1.15. |
4. In vitro ADME data.<br><br>a. A report (Powerpoint) for up to ten (10) compounds per Initially Selected DUB will be provided containing the data for hepatocyte and plasma stability, MDCK permeability, logD, solubility and protein binding, plus calculated physico-chemical parameters. |
5. Scale of chemistry delivery in order to determine whether a DUB target is chemically tractable:<br>a. 32 Piramal chemists will be deployed across 8 DUB targets for 3 months to generate ~40 compounds per Initially Selected DUB.<br>b. These compounds will be evaluated in primary biochemical, activity-based cellular an... |
Schedule B to Schedule 1.157 |
Stage 2 Minimum Deliverables |
1. Information and data to support AbbVie's Stage 2 Option decision. |
2. Data for in vitro assays conducted with the Mission Derived Compounds and the AbbVie Derived Compounds (i.e. Selected DUB biochemical SPS and IC50 screening, Agreed Assays selectivity panel (DSS), cellular TE, in vitro synuclein/tau clearance assays). The biochemical SPS and IC50 screening assays will be conducted o... |
3. Translational pharmacodynamic endpoint biomarker assay development data. |
4. Mechanism of Action (MOA) characterization data. |
5. Transfer of Selected DUB specific assays, reagents, protocols. |
6. Full DUB IC50 selectivity screen (for all DUBs with existing Suitable Biochemical Assay) on selected compounds. |
Mission Therapeutics Approved Subcontractors |
Piramal Enterprises Ltd<br>18 Pharmaceutical Special Economic Zone<br>PHARMEZ<br>Ahmedabad - 382110<br>GUJARAT<br>India |
Agility Health Tech Limited<br>Stevenage Bioscience Catalyst<br>Gunnels Wood Road<br>Stevenage, SG1 2FX<br>Great Britain |
Alderley Analytical Ltd<br>3G48 Biohub at Alderley Park<br>Alderley Edge<br>Macclesfield, SK10 4TG<br>Cheshire<br>Great Britain |
Aptuit<br>111 Innovation Drive<br>Milton Park<br>Abingdon, OX14 4RZ<br>Oxon<br>Great Britain |
Aragen Bioscince Inc<br>380 Woodview Avenue<br>Morgan Hill<br>CA 95037<br>United States |
BioTechne<br>19 Barton Lane<br>Abingdon Science Park<br>Abingdon<br>OX14 3NB |
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.