Bio-Energetic Sequencing - 5 Phases of Cellular Rejuvenation
$20M Series A

The First Dependency-Gated Framework for Systemic Age Reversal

Sequence matters more than compounds. We proved it mathematically.

6 Phases
120+ Year Ceiling
E→C→S→R→P Protocol
Explore the Science

The Dependency Chain

Each phase must be established before the next can activate. This is not a supplement stack—it is a state transition protocol grounded in control theory. The cellular system is modeled as a dynamical system where state variables evolve according to intervention inputs and temporal dynamics.

dx/dt = f(x, u, t)
y = h(x, u, t)
x = state vector (E, C, Sen, R, P, F)
u = interventions (compounds, timing, dose)
t = time (biological age, chronological progression)
y = observable biomarkers (NAD+, mTOR, p16, methylation)

These equations describe how cellular state evolves under interventions. Dependencies emerge from coupling terms in f(x,u,t)—autophagy activation depends on energetic capacity, senolysis safety depends on autophagic clearance, regeneration efficacy depends on cleared senescent burden.

E
Energy
C
Clearance
S
Senolysis
R
Regeneration
P
Programmatic

Five-Phase Protocol

Each phase builds the cellular capacity required for the next. Skipping a phase leads to intervention failure or creates pathological instability—rapamycin without energy causes metabolic collapse, senolytics without autophagy precipitate debris crisis, stem cell activation in damaged tissue increases cancer risk.

Phase 1: NAD+ and Sirtuin Activation
1

Energy

NAD+ Restoration

Restore mitochondrial capacity through NAD+ precursor supplementation and sirtuin activation. Without energetic sufficiency, cells cannot execute downstream processes such as autophagy, DNA repair, or senescent cell clearance.

Key Interventions:

  • NMN (500mg-1g) — NAD+ precursor
  • CoQ10 (200mg) — electron transport chain support
  • Apigenin (50mg) — CD38 inhibitor to prevent NAD+ degradation
NAD+ SIRT1 PGC-1α
Phase 2: Autophagy Activation
2

Clearance

Cellular Cleanup

Activate autophagy to clear damaged proteins, dysfunctional organelles, and accumulated cellular debris. This phase requires stable energetic capacity from Phase 1 to sustain the energy-intensive process of autophagic degradation and lysosomal recycling.

Key Interventions:

  • Rapamycin (5-10mg weekly) — mTOR inhibition, autophagy induction
  • Spermidine (1-5mg) — polyamine autophagy enhancer
  • Time-restricted eating — metabolic switch to promote autophagic flux
mTOR Autophagy ULK1
Phase 3: Senolytic Clearance
3

Senolysis

Senescent Removal

Selectively clear senescent cells that have accumulated due to chronic stress, DNA damage, and telomere attrition. This intervention is only safe after autophagy is active—clearing senescent cells without autophagic capacity to process debris results in inflammatory crisis and tissue damage.

Key Interventions:

  • Dasatinib + Quercetin — BCL-2 family inhibition for apoptosis induction
  • Fisetin (20mg/kg) — senolytic flavonoid
  • Pulsed protocols — intermittent dosing to minimize off-target effects
SASP p16 BCL-2
Phase 4: Stem Cell Regeneration
4

Regeneration

Tissue Renewal

Activate stem cell renewal and tissue regeneration pathways. This phase requires a cleared cellular environment—senescent cells secrete factors that inhibit stem cell function and increase cancer risk, making prior clearance essential for safe regenerative interventions.

Key Interventions:

  • Exosome therapy — regenerative signaling and paracrine effects
  • GDF11 modulation — systemic rejuvenation factor
  • Resistance training — mechanical stimulus for myogenic stem cell activation
Wnt Notch YAP/TAZ
Phase 5: Epigenetic Optimization
5

Programmatic

Epigenetic Reset

Shift epigenetic state toward youthful expression patterns through methylation optimization and histone modification. This final layer operates on gene expression programs—it is only effective after structural cellular renewal has been achieved through prior phases.

Key Interventions:

  • Alpha-ketoglutarate — TET enzyme cofactor for demethylation
  • TMG (trimethylglycine) — methyl donor for homocysteine metabolism
  • Vitamin D3 + K2 — epigenetic modulator and calcium regulation
DNAm TET Horvath

Computational Biology Platform

Every protocol decision is grounded in systematic computational analysis across peer-reviewed literature, pathway dependency mapping, and clinical translation. This is not trend-following or intuition—it is rigorous evidence synthesis and mathematical modeling.

Literature Synthesis

1,000+ Papers Analyzed

Systematic extraction of intervention data from peer-reviewed literature across Nature, Cell, Science, The Lancet, and specialized aging journals including Aging Cell, Rejuvenation Research, and GeroScience. Every claim is traced to primary source data with documented effect sizes, study populations, and methodological quality assessments.

PubMed Meta-Analysis Evidence Grading

Pathway Analysis

Dependency Mapping

Computational mapping of signaling cascade dependencies including NAD+/SIRT1/PGC-1α mitochondrial biogenesis, mTOR/AMPK/ULK1 autophagy regulation, and p53/p21/SASP senescence pathways. This analysis identifies which biological processes gate which downstream interventions, creating the empirical foundation for the dependency chain.

Coupling Matrix Phase Transitions Feedback Loops

Protocol Development

Clinical Translation

Translation of computational findings into implementable clinical protocols with precise dosing schedules, biomarker monitoring plans, phase-transition criteria, and safety gates. This includes individual variation modeling to account for metabolic heterogeneity, drug interactions, and baseline health status across patient populations.

Dosing Biomarkers Safety Gates

Research Foundation

The Bio-Energetic Sequencing Model is based on Principia Sanitatis by Mullo Saint, published through American Longevity Science. The framework is grounded in six theoretical papers and four peer-reviewed research studies demonstrating intervention sequencing effects and dependency violations.

Framework Papers

Complete theoretical foundation published through American Longevity Science:

Read All Framework Papers →

Published Research

Original research and clinical validation studies:

View All Research →

Why Dependencies Matter

Most longevity protocols fail because they ignore cellular state dependencies. Rapamycin without energetic sufficiency causes metabolic collapse through excessive ATP consumption. Senolytics without autophagic capacity precipitate a debris crisis as cellular fragments overwhelm clearance mechanisms. Stem cell activation in a senescent-rich environment increases cancer risk through exposure to pro-inflammatory, pro-proliferative SASP factors. The sequence is not optional—it is the mechanism of action.

$20M Series A

Building the first clinically-validated, dependency-aware longevity platform with Phase I/II trials, AI-driven personalization, and regulatory pathway development.

40%
Clinical Validation

Phase I/II trials, biomarker tracking, safety monitoring

25%
Platform Development

AI-driven personalization, state monitoring systems

20%
Regulatory

FDA pathway, compliance, clinical governance

10%
Category Leadership

Education, research publication, ecosystem growth

5%
Team

Core scientific and operational team expansion

The Longevity Ecosystem

Engineered Healthspan is part of a broader research and development ecosystem spanning personal protocols, theoretical frameworks, and peer-reviewed validation.

Mullo Saint

Personal research hub documenting longevity protocols, biohacking experiments, and self-quantification data. Serves as the empirical testbed for protocol development and individual variation modeling.

Visit mullosaint.com →

American Longevity Science

Theoretical foundation and mathematical frameworks including the Continuity Assurance Theorem, Bio-Energetic Sequencing Law, and Viable Zone Theory. Houses the complete six-paper framework series.

Visit americanlongevityscience.com →

Research Portal

Published studies demonstrating intervention sequencing effects, dependency violations, and clinical validation of the Bio-Energetic Sequencing Model across cellular and organismal models.

Visit research.mullosaint.com →