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Translational Biomarkers
Proof of Pharmacology
Proof of Biology
PK and Pharmaceutics
Physicochemical Properties
Intellectual Property
r/mu/d/mky/hu microsomal and hepatocyte stability
Plasma protein binding (r/mu/d/mky/hu)
CYP Inhibition Panel
Cellular Permeability
Rodent PK
Escalating dose rodent PK
Non-rodent PK
CYP phenotyping and induction and possible need to detect non-CYP metabolism and/or active transport
Human PK predictions
In Vitro Safety Pharmacology and Toxicology
hERG
Receptor selectivity panel
Mini AMES
In vitro and in vivo Micronucleus
PRECLINICAL DEVELOPMENT
Stage 3: Candidate Nomination to Candidate Selection and Initiation of GLP Toxicology Activities
Aim: To further evaluate and select two (2) optimal candidates to advance into preclinical development and initiate IND-enabling GLP toxicology for the lead molecule. When supported by data, candidate selection will include a frontrunner from one series and a back-up from a distinct chemical series
All Stage 3 Activities are the responsibility of Hotspot Therapeutics unless noted otherwise below.
Lead molecules (Stage 2 Deliverables) from both chemical series (and minimally one chemical series) will be tested in relevant in vitro and in vivo studies outlined below to support candidate nomination and candidate selection (Table 3). These studies will continue post-candidate selection to support PK/PD and human do...
Conduct Long term efficacy model
i. Spontaneous NZBxW/F1 mouse to be conducted to establish an efficacy ED50 <10mg/kg or an efficacious dose that scales to a reasonable efficacious human dose.
ii. Data generated should enable understanding of efficacy, target engagement (TE) and exposure response.
iii. PK/PD data to confirm preliminary safety margin calculated from CIA efficacy model and further support efficacious human dose projection.
iv. Hotspot Therapeutics accountable for generating data but potential to conduct long term efficacy model at AbbVie on approval of JGC.
Provision of non-GMP material for 2 species dose range finding (DRF) studies will be coordinated by Hotspot Therapeutics. Both AbbVie and Hotspot Therapeutics will contribute to the CMC work plan to be approved by the JGC (Appendix B) for DRF bulk delivery and enabling GMP manufacture.
In vivo safety will be assessed by Hotspot Therapeutics in rodent and non-rodent dose-range finding (DRF) studies to understand acute tolerability and high dose toxicokinetics (TK). Species selection will incorporate cross-species pharmacology and metabolic profiling data.
Conduct mechanistic PK and metabolism studies as needed, as described in Table 3 to support:
i. toxicology evaluation(s) such as potential of metabolite(s) to induce toxicity;
ii. to support refinement of human PK predictions including comparison of primary metabolic pathways and kinetics in animals and human in vitro, and;
iii. determine major metabolizing enzymes and/or transporters contributing to clearance and DDI potential of each candidate as perpetrator and victim.
A salt screen, if required based on compound(s) physiochemical properties, and polymorph screen will be carried out prior to manufacture of the non-GMP batch to support GLP tox studies. This non-GMP batch will also support initial ICH stability studies, and formulation development activities including Tech batch stabil...
Final agreement for the GMP Active Pharmaceutical Ingredient (API) manufacture will be approved by the JGC. The GMP API and Drug Product (DP) CDMO approval will be subject to the subcontractor provision in the Agreement. These activities will be initiated following Candidate Nomination and following provision of non-GM...
In parallel with detailed preclinical candidate selection studies (Table 3) with the most advanced current leads from each chemical series, a backup program will be initiated to identify additional candidate quality molecules for each chemical series, as approved by the JGC. The profile of the potential backup will be ...
Non-GMP Material synthesis
Following selection of a leading candidate and estimates based on the duration of GLP toxicology studies approved by the JGC at Candidate Selection, manufacture of sufficient non-GMP material will be initiated by Hotspot Therapeutics along with supporting studies on stability, impurity profile, etc as described in Appe...
Planning and initiation of IND-enabling GLP toxicology studies
Following selection of a leading candidate at Candidate Selection, GLP toxicology studies will be planned and initiated. Initiation in this case is defined as the start of the experimental or dosing phase of the first in vivo GLP toxicology study as detailed below in Table 3. It is anticipated that the duration of the ...
a. in vitro genetic toxicology studies (AMES and Micronucleus), in vivo micronucleus
b. in vitro phototoxicity (ONLY if needed based on absorption of light in UV range or photo-instability)
c. a single-dose cardiovascular telemetry study in non-rodents
d. respiratory and CNS rodent safety pharmacology assessments which may be incorporated into the repeat-dose toxicity study in rodents
e. a repeat-dose (likely 2-week with a 2-week post-dose recovery) toxicity study in one rodent species, and
f. a repeat-dose (likely 2-week with a 2-week post-dose recovery) toxicity study in one non-rodent species
The conduct of GLP toxicology studies will be the responsibility of Hotspot Therapeutics. All activities will be agreed in collaboration between both parties, in accordance with the regulatory requirements of AbbVie and with JGC approval. Timing of study conduct will also be coordinated with subsequent plans for 3-mont...
The Stage 3 Activities will be considered complete after the first animal is dosed in the first in vivo GLP toxicology study.
Stage 3 (Candidate Selection) Deliverables:
Deliverables to include analyzed data summaries on all compounds meeting candidate selection (CS) criteria for each activity performed in this stage and as requested by AbbVie, all analyzed and/or raw data for all compounds.
1. Two differentiated compounds (ideally one from each chemical series but minimally one compound from the lead series) selected that fulfils criteria outlined in Table 3 below.
2. Completion of rodent toxicity and non-rodent TK/tolerability (minimally 7-day repeat dose) plus at least one in vivo cardiovascular (CV) assessment that collectively provide acceptable safety profiles and defined therapeutic margins to progress to pre-first in human (FIH) GLP toxicology studies by Hotspot Therapeuti...
3. Understanding of potential on-target safety risks, responsibility of Hotspot Therapeutics.
4. Translational biomarker packages (discovered in Stage 2: Lead Optimization; Table 2) including delivery of candidate pharmacodynamic biomarkers with preclinical validation and evidence to support clinical proof of mechanism/pharmacology studies. The expectation is that these biomarkers will require further validatio...
Table 3. Stage 3 Pre-specified Criteria for Selection of a Development Candidate and Initiation of GLP Toxicology Studies
Inclusive of meeting Stage 1 and Stage 2 prespecified criteria listed in Tables 1 and 2
Activity / Assay
In Vitro and In Vivo preclinical studies
Long term efficacy model: Spontaneous NZBxW/F1 mouse
CV and Secondary pharmacodynamics
Rodent 7-day toxicity (dose range finding)
Non-rodent TK/tolerability study (dose range finding; minimally 7 day)
In vitro ADME study in microsomes and hepatocytes
De-risk circulating metabolites identified for either pharmacological activity and/or bioactivation
PK and metabolism
GLP bioanalytical assay
Initiation of the dosing phase of the 1st repeat dose GLP toxicology study
Stage 4: IND Activities
Aim: To generate and assemble a comprehensive nonclinical data package (toxicology and CMC) to support an IND/CTA submission.
All Stage 4 Activities are the responsibility of Hotspot Therapeutics unless otherwise noted below.
1. Provision of top line GLP toxicology data that will subsequently be part of IND-ready package. HotSpot Therapeutics will initially deliver a comprehensive set of top line data from all GLP studies, which includes completion of all data analysis of toxicology, toxicokinetic, and clinical and anatomic/histopathology p...
2. The integrated top line data will subsequently be used in preparation of the IND-ready package and will be the responsibility of Hotspot Therapeutics. These accountabilities will include: (1) transfer of data in suitable format including SEND-ready format for GLP toxicology and toxicokinetic data, and; (2) provision...
3. GMP material synthesis
If needed, Hotspot Therapeutics will synthesize radiolabelled material (14C or 3H) to support mechanistic ADME-Tox studies pre-FIH. The requirement for this will be approved by the JGC.
Subsequently GMP material will be manufactured by Hotspot Therapeutics to support Phase 1 studies with supporting studies as described in Appendix B.
To support the wide dose range typically studied in the first in human single ascending dose study, a simple oral dosing suspension/solution or powder in capsule formulation will likely be used dependent upon emergent information about physical/chemical properties of the molecule in combination with oral bioavailabilit...
The Stage 4 Activities will be considered complete once the first Phase 1 clinical study is permitted to proceed following submission of the IND/CTA package, as applicable, to the applicable Regulatory Authority.
Stage 4 Deliverables:
Delivery of top line data from the IND-enabling GLP toxicology package (including histopathology data and slides) including completion of all data analysis and HotSpot Therapeutics management review of data to enable creation of an integrated summary for AbbVie's review from the listing of studies outlined in Table 4 b...
Subsequently, finalization of study reports, SEND datasets and integrated summary documents to support IND/CTA submission will be responsibility of HotSpot Therapeutics and as described in Appendix C.
Manufacture of sufficient non-GMP and GMP material by Hotspot Therapeutics as described in Appendix B.
Table 4. Stage 4 Pre-specified Criteria for Completion of GLP Toxicology Studies and Delivery of an IND-enabling Data Package
Inclusive of meeting Stage 1, Stage 2, and Stage 3 prespecified criteria listed in Tables 1, 2, and 3.
Activity / Assay
Completion of GLP Toxicology Studies and Delivery of IND-enabling Data Package
Delivery of top line data from the full listing of GLP toxicology studies for AbbVie to review including the following studies: a. In vitro genetic toxicology studies (AMES and Micronucleus); in vivo micronucleus b. In vitro phototoxicity (ONLY if needed based on absorption of light in UV range or photo-instability) c....
Post-IND/Post-Stage 4 Activities
Aim: To generate 13-week toxicology and Phase 1 SAD and MAD clinical data to support assembly of the Final Data Package for AbbVie Opt-in.
13-week GLP Toxicology
Initiation of 13-week GLP toxicology study and delivery of top line data from the 13-week GLP toxicology study in one rodent and one non-rodent species
CLINICAL DEVELOPMENT
Phase 1 Set Up
Preparation for First in Human (FIH) study will be managed and approved by the JGC as outlined below:
1. Bioanalytical Method and Pharmacodynamic Biomarkers
Development and validation of bioanalytical methods according to FDA/EMA Regulatory Guidelines for use in the FIH study are the responsibility of Hotspot Therapeutics. Development of potential pharmacodynamic markers with relevant preclinical validation for use in the FIH study are the responsibility of Hotspot Therape...
2. General Investigational Plan and FIH study (SAD/MAD in Healthy Volunteers; HVs)
The Target Product Profile and General Investigational Plan to deliver on the profile are the responsibility of Hotspot Therapeutics. Details will be shared to and approved by the JGC.