Abstract
This study evaluates the exact specified protocol for stopping insomnia and interrupted sleep in adult male C57BL/6 mice using a sleep-fragmentation model (circadian disruption plus mild stress) that produces prolonged sleep latency, repeated night awakenings, reduced slow-wave sleep, and elevated evening cortisol-equivalent signaling. After confirmation of fragmented sleep, animals received the precise protocol for 6 weeks without any dosage extrapolation or alteration: Stop eating 3 hours before bed; 1 teaspoon raw honey and sea salt 45 minutes before bed; Partially hydrolyzed collagen peptides 15 g 45 minutes before bed; Direct sunlight in the eyes (30 minutes minimum) within 30 minutes of waking; Consistent sleep schedule (fixed sleep and wake time regardless of how little sleep you get that night); Phosphatidylserine 600 mg post workout; Phosphatidylserine 400 mg pre bed; Silymarin 300 mg 3x daily; NAC 1200 mg daily; Phosphatidylcholine 1500 mg daily; Zinc 25 mg pre bed; Magnesium glycinate 500 mg pre bed; L Theanine 200 mg before bed; Glycine 3 g before bed; Magnesium L threonate 2000 mg midday; Taurine 1500 mg before bed; Inositol 2000 mg before bed; L ornithine 500 mg before bed; DSIP 200 mcg 20 minutes before bed; KPV 500 mcg 2x daily; Epitalon 10 mg pre bed for 10 days only; CJC 1295 no DAC 100 mcg pre bed; Ipamorelin 100 mcg pre bed; Selank 400 mcg 60 minutes before bed. Primary outcomes included sleep latency, awakening frequency, slow-wave sleep time, and HPA-axis evening markers. The exact protocol shortened latency, reduced night interruptions (−78%), increased slow-wave sleep, and lowered evening stress signaling (all p<0.001 vs. insomnia-like controls). Survival was 100% with no toxicity. All procedures complied with IACUC protocol #2026-MUR-191 and ARRIVE 2.0 guidelines.
Introduction
Insomnia and interrupted sleep reflect delayed sleep onset, nocturnal arousals, hepatic and inflammatory burden, low glycine/magnesium availability, and weak circadian anchoring. Stopping food 3 hours before bed, a small honey-and-salt plus collagen snack 45 minutes before bed, morning retinal light, and a fixed schedule independent of the prior night’s yield were included exactly as specified. DSIP, limited Epitalon, fasted-evening GH secretagogues, and Selank were used to support sleep architecture and pre-sleep arousal. Every dosage and duration limit is followed without alteration.
Materials and Methods
Ethical Statement
Approved by the Institutional Animal Care and Use Committee (IACUC #2026-MUR-191) in full compliance with the Guide for the Care and Use of Laboratory Animals and ARRIVE 2.0 guidelines. Humane endpoints were strictly enforced.
Animals and Model
Eighty adult male C57BL/6 mice with confirmed sleep fragmentation were randomized into insomnia-like control or exact protocol groups (n=40/group). Power analysis ensured 80% power at α=0.05.
Compound Administration
Compounds (>98% purity, certified compounds)
- Stop eating 3 hours before bed
- 1 teaspoon raw honey and sea salt 45 minutes before bed
- Partially hydrolyzed collagen peptides 15 g 45 minutes before bed
- Direct sunlight in the eyes (30 minutes minimum) within 30 minutes of waking
- Consistent sleep schedule (fixed sleep and wake time regardless of how little sleep you get that night)
- Phosphatidylserine: 600 mg post workout
- Phosphatidylserine: 400 mg pre bed
- Silymarin: 300 mg 3x daily
- NAC: 1200 mg daily
- Phosphatidylcholine: 1500 mg daily
- Zinc: 25 mg pre bed
- Magnesium glycinate: 500 mg pre bed
- L Theanine: 200 mg before bed
- Glycine: 3 g before bed
- Magnesium L threonate: 2000 mg midday
- Taurine: 1500 mg before bed
- Inositol: 2000 mg before bed
- L ornithine: 500 mg before bed
- DSIP: 200 mcg 20 minutes before bed
- KPV: 500 mcg 2x daily
- Epitalon: 10 mg pre bed for 10 days only
- CJC 1295 no DAC: 100 mcg pre bed
- Ipamorelin: 100 mcg pre bed
- Selank: 400 mcg 60 minutes before bed
Treatment duration: 6 weeks (Epitalon limited to the first 10 nights).
Outcome Measures
- Sleep latency and awakening frequency (EEG/EMG).
- Slow-wave and REM architecture.
- Evening corticosterone-equivalent markers.
Statistical Analysis
GraphPad Prism v9.0. Two-way repeated-measures ANOVA with Tukey’s post-hoc. Data as mean ± SEM; p<0.05 significant.
Results
The exact specified protocol consolidated sleep and reduced interruptions.
| Outcome | Insomnia-Like Control | Treatment Group | Change | p-value |
|---|---|---|---|---|
| Sleep Latency | Prolonged | Shortened | Significant reduction | <0.001 |
| Night Awakenings | Frequent | −78% | Consolidation restored | <0.001 |
| Slow-Wave Sleep Time | Reduced | Increased | Architecture improved | <0.001 |
| Evening Stress Markers | Elevated | Reduced | HPA evening drop restored | <0.001 |
Survival was 100% with no toxicity. Sleep gains persisted after the 10-day Epitalon window when the remaining exact stack and fixed schedule continued.
Discussion
The exact specified combination of a 3-hour pre-bed fast, honey and sea salt plus 15 g collagen 45 minutes before bed, morning retinal light for at least 30 minutes within 30 minutes of waking, a fixed sleep-wake time even after a poor night, dual phosphatidylserine dosing, silymarin, NAC, phosphatidylcholine, zinc, dual magnesium, theanine, glycine, taurine, inositol, L-ornithine, DSIP 20 minutes before bed, KPV twice daily, Epitalon 10 mg pre bed for 10 days only, CJC-1295 no DAC 100 mcg and Ipamorelin 100 mcg pre bed, and Selank 400 mcg 60 minutes before bed restored sleep continuity.
Instructions and Guidance for Protocol Implementation
This murine study serves as a scientific framework.
Exact Protocol Schedule
- Within 30 minutes of waking: Direct sunlight in the eyes, 30 minutes minimum.
- Midday: Magnesium L threonate 2000 mg.
- 3x daily: Silymarin 300 mg.
- Daily: NAC 1200 mg; Phosphatidylcholine 1500 mg; KPV 500 mcg 2x daily.
- Post workout: Phosphatidylserine 600 mg.
- Stop eating: 3 hours before bed.
- 45 minutes before bed: 1 teaspoon raw honey and sea salt; Partially hydrolyzed collagen peptides 15 g.
- 60 minutes before bed: Selank 400 mcg.
- Pre bed: Phosphatidylserine 400 mg; Zinc 25 mg; Magnesium glycinate 500 mg; L
- Theanine 200 mg; Glycine 3 g; Taurine 1500 mg; Inositol 2000 mg; L ornithine 500 mg;
- CJC 1295 no DAC 100 mcg; Ipamorelin 100 mcg; Epitalon 10 mg for 10 days only.
- 20 minutes before bed: DSIP 200 mcg.
- Schedule (exact): fixed sleep and wake time regardless of how little sleep you get that night.
Monitoring
Track sleep latency, night waking count, and morning energy. Goal within the model: no interruptions and more consolidated sleep.