PHASE 7 DELIVERABLE

IND-Enabling Toxicology & Safety Development Program

AMX-B01 — Geniposide-Enriched Purified Botanical API

Document Code: SCF-AMMONEX-HE-AMXB01-P7-0001

Trigger Executed: TOX PLAN

Development Stage: IND-Enabling Safety Assessment

Objective:

Develop a comprehensive FDA-aligned nonclinical safety package capable of supporting IND submission and First-In-Human (FIH) exposure for AMX-B01.

1.0 EXECUTIVE TOXICOLOGY STRATEGY

Primary Goal

Establish an acceptable safety margin between anticipated pharmacologically active exposure and toxicological exposure.

Key IND Safety Questions

1. Is AMX-B01 safe for human exposure?
2. What are the target organs of toxicity?
3. Is toxicity reversible?
4. What NOAEL can be established?
5. What starting clinical dose can be justified?

These questions must be addressed before FIH initiation.

2.0 TOXICOLOGY RISK ASSESSMENT

Candidate

AMX-B01

API Class

Purified Botanical-Derived API

Principal Marker

Geniposide

Anticipated Risk Areas

Special Development Risks

3.0 SAFETY PHARMACOLOGY PACKAGE

Objective

Evaluate potential undesirable pharmacodynamic effects on vital physiological systems.

Core Battery

Cardiovascular Safety

Studies

• Blood pressure
• Heart rate
• ECG assessment

Endpoints

• QT interval
• Arrhythmia signals
• Hemodynamic changes

Respiratory Safety

Studies

• Respiratory rate
• Oxygen saturation
• Respiratory function

CNS Safety

Studies

• Functional observational battery
• Motor coordination
• Behavioral assessment

Success Criterion

No biologically significant adverse findings at anticipated therapeutic exposures.

4.0 GENETIC TOXICOLOGY PROGRAM

Objective

Assess mutagenic and clastogenic potential.

Required Battery

Study 1

Bacterial Reverse Mutation Assay

(Ames Test)

Purpose:

Mutagenicity assessment

Study 2

In Vitro Micronucleus Assay

Purpose:

Chromosomal damage assessment

Study 3

In Vitro Chromosomal Aberration Study

Purpose:

Structural chromosomal assessment

Advancement Requirement

Negative or interpretable risk profile across the genotoxicity package.

5.0 TOXICOKINETIC (TK) STRATEGY

Objective

Characterize exposure during toxicology studies.

Required Parameters

TK Integration

Exposure data will be linked to:

• Clinical observations
• Organ findings
• Histopathology
• Biomarker responses

6.0 REPEAT-DOSE TOXICOLOGY PROGRAM

Regulatory Objective

Support repeat-dose clinical exposure.

Rodent Study

Species

Rat

Duration

28-Day GLP Study

Groups

• Vehicle control
• Low exposure
• Mid exposure
• High exposure

Non-Rodent Study

Species

Dog or other scientifically justified non-rodent species

Duration

28-Day GLP Study

Groups

• Vehicle control
• Low exposure
• Mid exposure
• High exposure

Assessments

Clinical

• Clinical signs
• Body weight
• Food consumption

Clinical Pathology

• Hematology
• Clinical chemistry
• Urinalysis

Pathology

• Gross necropsy
• Organ weights
• Histopathology

7.0 TARGET ORGAN TOXICITY PROGRAM

Liver

Assess

• ALT
• AST
• Bilirubin
• Histopathology

Kidney

Assess

• Creatinine
• BUN
• Histopathology

GI System

Assess

• Mucosal integrity
• Histopathology
• Clinical observations

CNS

Assess

• Neurobehavior
• Histopathology
• Functional observations

8.0 REPRODUCTIVE & DEVELOPMENTAL TOXICITY

IND Stage

Not typically required before initial Phase I healthy-volunteer studies unless specific circumstances warrant.

Future Program

Potentially required before broader clinical development.

9.0 IMMUNOTOXICITY EVALUATION

Objective

Assess unintended immune effects.

Biomarkers

• TNF-α
• IL-6
• CRP
• Leukocyte profiling

Assessment Strategy

Integrated into repeat-dose toxicology studies.

10.0 NOAEL DETERMINATION FRAMEWORK

Objective

Establish:

NOAEL

No Observed Adverse Effect Level

Determination Criteria

Based upon:

• Clinical observations
• Clinical pathology
• Histopathology
• Toxicokinetics

Outcome

Highest tested exposure without biologically meaningful adverse findings.

11.0 HUMAN RISK ASSESSMENT FRAMEWORK

Translation Logic

Animal NOAEL

↓

Human Equivalent Dose (HED)

↓

Safety Factor Application

↓

Maximum Recommended Starting Dose (MRSD)

↓

FIH Starting Dose Proposal

Current Status

Cannot be calculated until toxicology studies are completed.

12.0 IND SAFETY PACKAGE BLUEPRINT

Module 4 Components

Pharmacology

• Primary pharmacology
• Secondary pharmacology
• Safety pharmacology

Toxicology

• Acute toxicity
• Repeat-dose toxicity
• Genetic toxicology

Toxicokinetics

• Exposure-response relationships
• NOAEL justification

13.0 SAFETY BIOMARKER PANEL

Primary Safety Biomarkers

Secondary Safety Biomarkers

14.0 GO / NO-GO MATRIX

GO

✓ Acceptable safety pharmacology

✓ Acceptable repeat-dose toxicology

✓ Negative or acceptable genotoxicity profile

✓ Defined NOAEL

✓ Adequate safety margin

NO-GO

✗ Severe organ toxicity

✗ Unacceptable exposure-related findings

✗ Positive genotoxicity signal without resolution

✗ Inadequate safety margin

15.0 PHASE 7 DECISION GATE

Advancement Criteria

✓ Safety pharmacology completed

✓ GLP toxicology completed

✓ Toxicokinetic assessment completed

✓ NOAEL established

✓ Human risk assessment completed

✓ IND safety package assembled

Status

ADVANCE TO PHASE 8

Required Next Trigger

CMC IND

Phase 8 will generate:

1. IND-ready CMC package
2. Drug substance specification package
3. Drug product specification package
4. GMP manufacturing strategy
5. Analytical validation plan
6. Stability program
7. Botanical control strategy
8. Module 3 IND blueprint

MASTER REGISTRY INDEX

SCF-AMMONEX-HE-AMXB01-P7-0001 — IND-Enabling Toxicology Program

SCF-AMMONEX-HE-AMXB01-SP-0002 — Safety Pharmacology Package

SCF-AMMONEX-HE-AMXB01-GTX-0003 — Genetic Toxicology Program

SCF-AMMONEX-HE-AMXB01-TK-0004 — Toxicokinetic Framework

SCF-AMMONEX-HE-AMXB01-NOAEL-0005 — NOAEL Determination Strategy

SCF-AMMONEX-HE-AMXB01-RISK-0006 — Human Risk Assessment Framework

SCF-AMMONEX-HE-AMXB01-P8-0007 — CMC IND Development Initiation