NA Epitalon

Research Overview

N-Acetyl Epitalon serves as a valuable research tool for investigating whether peptide modifications enhance biological activity and pharmacokinetic properties in laboratory settings. This acetylated derivative of Epitalon was developed to address potential limitations of unmodified short peptides, specifically susceptibility to aminopeptidase degradation at the N-terminus. The addition of an acetyl group (COCH₃) to the alpha-amino group of the N-terminal alanine creates a modified peptide with potentially altered stability and cellular interaction characteristics.

The rationale for N-acetylation stems from established peptide chemistry principles. Unmodified peptides with free N-terminal amino groups serve as substrates for aminopeptidases, enzymes that sequentially cleave amino acids from the N-terminus. Acetylation blocks this degradation pathway, potentially extending peptide half-life in biological systems. Research applications examine whether this modification translates to enhanced biological effects, improved cellular uptake, or altered mechanism of action compared to standard Epitalon.

N-Acetyl Epitalon research demonstrates the broader principle of structure-activity relationship studies in peptide science. By comparing acetylated and non-acetylated versions, researchers can investigate the importance of N-terminal structure, charge distribution, and metabolic stability to overall biological activity. Studies examine effects on telomerase activity, cellular senescence markers, and neuroendocrine function to determine if acetylation provides research advantages.

Molecular Characteristics

Complete Specifications:

• CAS Registry Number: NOT ASSIGNED (synthetic modified peptide)
• Molecular Weight: 349.3 Da (42 Da higher than Epitalon due to acetyl group)
• Molecular Formula: C₁₆H₂₄N₄O₁₀
• Amino Acid Sequence: N-Ac-Ala-Glu-Asp-Gly (acetylated AEDG)
• Modification: N-terminal acetylation (acetyl group: COCH₃)
• Peptide Classification: Synthetic modified tetrapeptide
• Appearance: White to off-white lyophilized powder
• Solubility: Water, bacteriostatic water, phosphate buffered saline

The N-acetyl modification adds 42 Da to the molecular weight compared to unmodified Epitalon (307.3 Da). This acetyl group (COCH₃) replaces the positive charge on the N-terminal amino group with a neutral acetyl moiety, altering the peptide’s charge distribution and hydrophobicity. The modification reduces overall positive charge at physiological pH, potentially affecting cellular interactions, membrane permeability, and receptor binding characteristics. These structural differences form the basis for comparative research studies.

Pharmacokinetic Profile in Research Models

N-Acetyl Epitalon pharmacokinetic characterization focuses on comparing properties to unmodified Epitalon:

Absorption and Bioavailability:

• Hypothesized enhanced stability to aminopeptidase degradation
• Potential improved membrane permeability due to reduced charge
• Multiple administration routes under investigation
• Comparative bioavailability studies with standard Epitalon
• Oral bioavailability potentially improved vs. unmodified form

Distribution and Elimination:

• Plasma half-life comparison to standard Epitalon (hypothesis: extended)
• Tissue distribution patterns under investigation
• Metabolic stability assessment in plasma and tissue homogenates
• Renal clearance characteristics
• Cellular uptake and retention studies

Biological Activity Timeline:

• Comparative time-course studies with Epitalon
• Assessment of sustained vs. acute effects
• Telomerase activation kinetics and duration
• Dose-response relationships compared to unmodified form

Research protocols include direct head-to-head comparisons with standard Epitalon using identical experimental conditions to isolate effects attributable specifically to the acetyl modification.

Research Applications

Comparative Peptide Modification Studies

N-Acetyl Epitalon serves as an ideal research tool for structure-activity relationship investigations:

• Modification Impact Research: Direct comparison of biological activities between acetylated and non-acetylated versions
• Stability Enhancement Studies: Investigation of enzymatic resistance and metabolic stability improvements
• Membrane Permeability Research: Analysis of cellular uptake differences based on charge and hydrophobicity modifications
• Structure-Function Relationship: Examination of how N-terminal modifications affect receptor interactions and biological outcomes
• Pharmacokinetic Optimization: Research on how chemical modifications alter absorption, distribution, and elimination

Research protocols employ side-by-side comparisons in identical assay systems, allowing isolation of modification-specific effects.

Telomere Biology and Telomerase Research

Like standard Epitalon, N-Acetyl Epitalon research examines telomere-related mechanisms:

• Telomerase Activity Studies: Investigation of TERT expression and enzyme activity with modified peptide
• Comparative Efficacy Research: Assessment of whether acetylation enhances telomerase activation
• Telomere Length Dynamics: Longitudinal studies comparing telomere elongation between forms
• Cellular Senescence Research: Examination of replicative senescence delay with modified peptide
• Mechanism Investigation: Research on whether acetylation affects signaling pathways upstream of telomerase

Studies utilize TRAP assays, qPCR telomere length measurement, senescence-associated β-galactosidase staining, and cell proliferation assays comparing both peptide forms.

Enhanced Stability and Bioavailability Research

Key research focus examines whether acetylation provides practical advantages:

• Enzymatic Resistance Studies: In vitro stability testing in presence of aminopeptidases and peptidases
• Plasma Stability Research: Time-course degradation studies in biological matrices
• Cellular Uptake Enhancement: Comparative cellular internalization studies using labeled peptides
• Bioavailability Investigation: Assessment of systemic exposure following various administration routes
• Tissue Penetration Research: Analysis of tissue distribution and target organ accumulation

Experimental approaches include peptide stability assays in serum/plasma, cellular uptake quantification using mass spectrometry or radiolabeling, and pharmacokinetic profiling in animal models.

Neuroendocrine and Pineal Function Research

Research extends to neuroendocrine applications similar to standard Epitalon:

• Melatonin Production Studies: Investigation of pineal gland function with acetylated peptide
• Circadian Rhythm Research: Analysis of circadian clock gene modulation
• Blood-Brain Barrier Penetration: Studies examining whether acetylation affects CNS bioavailability
• Pinealocyte Function Research: Direct effects on pineal cell culture systems
• Comparative Neuroendocrine Effects: Head-to-head comparison with standard Epitalon

Laboratory studies utilize pineal gland explants, melatonin immunoassays, circadian behavior monitoring, and brain penetration studies.

Cellular Aging and Longevity Mechanism Research

Research investigates anti-aging mechanisms with the modified peptide:

• Age-Related Gene Expression: Research on longevity-associated gene modulation
• Oxidative Stress Response: Studies on antioxidant enzyme expression and oxidative damage markers
• Mitochondrial Function: Investigation of mitochondrial biogenesis and respiratory capacity
• DNA Repair Mechanisms: Analysis of DNA damage response and repair pathway activation
• Cellular Maintenance Pathways: Research on autophagy, proteostasis, and cellular quality control

Experimental models include comparative studies in young vs. aged systems, accelerated aging models, and longitudinal biomarker assessments.

Peptide Chemistry and Formulation Research

N-Acetyl Epitalon provides research opportunities in peptide science:

• Chemical Modification Effects: Investigation of how N-terminal modifications affect peptide properties
• Formulation Stability: Research on long-term stability in various formulation conditions
• Aggregation Behavior: Studies on peptide aggregation tendencies and prevention strategies
• Solution Behavior: Analysis of pH stability, solubility, and solution phase characteristics
• Delivery System Research: Investigation of peptide incorporation into delivery vehicles

Research employs analytical chemistry techniques including HPLC method development, forced degradation studies, aggregation analysis by size-exclusion chromatography, and formulation stability studies.

Laboratory Handling and Storage Protocols

Lyophilized Powder Storage:

• Store at -20°C to -80°C in original sealed vial
• Protect from light exposure and moisture
• Desiccated storage environment essential
• Stability data available for 12+ months at -20°C
• Acetyl modification may provide enhanced stability vs. unmodified Epitalon

Reconstitution Guidelines:

• Reconstitute with sterile water, bacteriostatic water (0.9% benzyl alcohol), or appropriate buffer
• Add solvent slowly down vial side to minimize foaming
• Gentle swirling motion recommended (avoid vigorous shaking)
• Allow complete dissolution before use (typically 1-3 minutes)
• Final pH should be 6.5-7.5 for optimal stability
• Calculate concentration based on peptide content (typically 80-85% by weight)

Reconstituted Solution Storage:

• Short-term storage: 4°C for up to 7 days (with bacteriostatic water)
• Long-term storage: -20°C to -80°C in single-use aliquots
• Single-use aliquots recommended to avoid freeze-thaw degradation
• Avoid repeated freeze-thaw cycles (maximum 2 cycles)
• Use protein low-binding tubes to minimize adsorption
• Compare stability to standard Epitalon under identical conditions

Quality Assurance and Analytical Testing

Each N-Acetyl Epitalon batch undergoes comprehensive analytical characterization:

Purity Analysis:

• High-Performance Liquid Chromatography (HPLC): ≥98% purity
• Reversed-phase HPLC with UV detection at 214-220nm
• Acetylated peptide elutes differently than unmodified form
• Multiple peak integration for accurate purity determination

Structural Verification:

• Electrospray Ionization Mass Spectrometry (ESI-MS): Confirms molecular weight 349.3 Da
• MS/MS fragmentation confirms acetyl modification presence
• Amino acid analysis: Verifies base sequence (Ala-Glu-Asp-Gly)
• Peptide content determination: Quantifies actual content (typically 80-85%)
• Acetylation confirmation via N-terminal sequencing or MS analysis

Contaminant Testing:

• Bacterial endotoxin: <5 EU/mg (LAL method)
• Heavy metals: Below detection limits per USP standards
• Residual solvents: TFA, acetonitrile within acceptable limits
• Water content: Karl Fischer titration (<8%)
• Unmodified Epitalon: Typically <1% if present

Documentation:

• Certificate of Analysis (COA) provided with each batch
• Acetylation verification included in analytical data
• Third-party verification available upon request
• Stability data for recommended storage conditions
• Batch-specific QC results traceable by lot number

Research Considerations

Experimental Design Factors:

Researchers should consider several factors when designing N-Acetyl Epitalon experiments:

1. Comparative Studies: Include standard Epitalon as direct comparison to isolate acetylation effects. Use identical experimental conditions, concentrations, and time points.

2. Concentration Adjustments: Account for molecular weight difference (349.3 Da vs. 307.3 Da) when comparing molar concentrations between forms.

3. Model System Selection: Choose experimental systems that allow meaningful comparison of stability and activity differences.

4. Controls: Include both vehicle controls and standard Epitalon controls in all experiments.

5. Pharmacokinetic Considerations: Design studies to specifically test hypothesized stability and bioavailability advantages.

6. Mechanism Investigation: Examine whether acetylation affects upstream signaling pathways differently than unmodified form.

Modification Impact Assessment:

Key research questions regarding N-acetylation include:

• Enhanced Stability: Does acetylation significantly extend peptide half-life in biological matrices?
• Improved Bioavailability: Does reduced charge improve membrane permeability and cellular uptake?
• Maintained Activity: Does modification preserve or enhance biological activity?
• Altered Mechanism: Does acetylation change receptor interactions or signaling pathways?
• Practical Advantages: Do stability improvements translate to enhanced biological effects?

Research should employ quantitative methods to assess these questions including stability assays, pharmacokinetic studies, and comparative efficacy assessments.

Compliance and Safety Information

Regulatory Status:
N-Acetyl Epitalon is provided as a research chemical for in-vitro laboratory studies and preclinical research only. This product has not been approved by the FDA for human therapeutic use, dietary supplementation, or medical applications.

Intended Use:

• In-vitro cell culture studies
• In-vivo preclinical research in approved animal models
• Comparative peptide modification research
• Structure-activity relationship studies
• Academic and institutional research applications

NOT Intended For:

• Human consumption or administration
• Therapeutic treatment or diagnosis
• Dietary supplementation or anti-aging products
• Veterinary therapeutic applications without oversight
• Any medical or clinical applications

Safety Protocols:

• Use appropriate personal protective equipment
• Handle in well-ventilated areas or biosafety cabinet
• Follow institutional biosafety guidelines
• Dispose of waste per local regulations
• Consult MSDS for additional safety information

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