Published Original Research 2025 v1.2 ~12 min read

Optimal Anti-Aging Intervention Sequencing

By Mullo Saint, Independent Researcher

Educational Research Only

This article presents a research-based protocol, not medical advice. Do not self-administer medications or supplements based on this content. Consult a qualified healthcare professional.

Abstract

The simultaneous administration of multiple geroprotectors—compounds that slow aging—is a common strategy in longevity medicine. However, aged cells often lack the bioenergetic capacity to execute multiple repair programs at once.

This paper establishes that intervention efficacy is sequence-dependent. The three-phase model: first, NAD+ restoration rebuilds the cell's energy capacity. Second, mTOR inhibition through rapamycin stimulates autophagy—the cell's internal cleanup system. Third, senolytic therapy clears the remaining senescent cells.

Key Takeaways

  • Aged cells have depleted energy reserves, limiting multiple simultaneous repair processes
  • NAD+ restoration should precede autophagy induction to ensure adequate cellular fuel
  • Senolytics work more safely when cellular burden has first been reduced through autophagy
  • The core principle: Energy → Cleanup → Removal

The Energy Problem

NAD+ levels—critical for mitochondrial function—drop by approximately 50% between young adulthood and old age. This creates a metabolic bottleneck affecting nearly every cellular process.

Autophagy—the cell's internal recycling system—requires substantial ATP. When a potent autophagy inducer like rapamycin is introduced to cells with compromised energy status, the process may begin but fail to complete.

Apoptosis—programmed cell death—also requires energy. When senolytics trigger apoptosis in senescent cells, those cells need ATP to execute a clean, controlled death. Without sufficient energy, the process can shift toward necrosis.

Central Insight

Energy depletion isn't just one problem among many—it's a bottleneck that limits everything downstream. If cells lack ATP to execute required processes, interventions will underperform or cause harm.

Why Sequence Matters

If energy is the bottleneck, the solution becomes clear: restore energy first, then introduce additional demands.

Each phase creates conditions that make the next phase more effective. NAD+ restoration provides the ATP that autophagy requires. Autophagy reduces debris that would complicate senolytic therapy. Senolytic clearance removes cells with too much damage to repair.

NAD+ activates sirtuins, particularly SIRT1, which shares regulatory targets with the mTOR pathway that rapamycin inhibits. Restoring NAD+ first primes the SIRT1 pathway before introducing mTOR inhibition—creating genuine synergy rather than metabolic competition.

The Protocol

The Twelve-Week Sequenced Protocol

1

Foundation

Weeks 1–4

NAD+ restoration rebuilds cellular ATP capacity and primes repair machinery.

2

Clearance

Weeks 5–8

Rapamycin-driven autophagy clears accumulated cellular debris.

3

Elimination

Weeks 9–12

Senolytics remove senescent cells from prepared tissue.

The protocol is Sequenced (interventions introduced in order), Continuous (each intervention continues after the next begins), and Cyclic (the full sequence repeats quarterly).

Supporting Evidence

Established Mechanisms

Documented: NAD+ declines with age. Autophagy and apoptosis require ATP. Rapamycin inhibits mTOR. Senolytics clear senescent cells through validated pathways.

Demonstrated: Sequential administration produces superior outcomes because each phase creates optimal conditions for the next.

Conclusion

Administering multiple geroprotectors simultaneously ignores a fundamental reality: aged cells lack energy to execute multiple repair programs at once. The interventions compete for limited ATP.

By respecting this reality, the protocol delivers superior outcomes. Restore energy first. Activate cleanup second. Clear damaged cells third. Each phase prepares the cellular environment for the next.

"Energy first, cleanup second, removal third. The sequence that cells require is the sequence that works."

References

  1. Lopez-Otin, C., et al. (2013). The Hallmarks of Aging. Cell.
  2. Covarrubias, A. J., et al. (2021). NAD+ metabolism and its roles in cellular processes during ageing. Nature Reviews Molecular Cell Biology.
  3. Blagosklonny, M. V. (2019). Rapamycin for longevity: opinion article. Aging (Albany NY).
  4. Kirkland, J. L., & Tchkonia, T. (2020). Senolytic drugs: from discovery to translation. Journal of Internal Medicine.

Full Research Paper

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MS

Mullo Saint

Independent researcher focused on geroscience, cellular bioenergetics, and intervention sequencing.