NCT04325308

Early Protein Supplementation in Extremely Preterm Infants Fed Human Milk

Active, enrolled NA Results posted Last updated 23 September 2025

What this trial tests

NA trial testing Protein-enriched human milk diet in Prematurity; Extreme in 150 participants. Participants enrolled and being followed up; not accepting new ones.

Timeline

13 August 2020

Primary endpoint
2 January 2023

30 September 2026

Quick facts

Lead sponsor	University of Alabama at Birmingham
Phase	NA
Status	Active, enrolled
Study type	INTERVENTIONAL
Allocation	randomized
Design	parallel
Masking	quadruple
Primary purpose	prevention
Enrollment	150
Start date	13 August 2020
Primary completion	2 January 2023
Estimated completion	30 September 2026
Sites	1 location across United States

Drugs / interventions tested

• Protein-enriched human milk diet
• Usual human milk diet

Conditions studied

• Prematurity; Extreme — all drugs for Prematurity; Extreme →
• Feeding Disorder Neonatal — all drugs for Feeding Disorder Neonatal →
• Breast Milk Expression — all drugs for Breast Milk Expression →
• Growth Failure — all drugs for Growth Failure →

Sponsor

University of Alabama at Birmingham

Who can join

Adults 1 Day to 4 Days, any sex, with Prematurity; Extreme or Feeding Disorder Neonatal. Patients with the condition only — healthy volunteers not accepted.

Results — posted to ClinicalTrials.gov

Per-arm endpoint measurements with 95% confidence intervals where reported. Source: trial results section.

Fat-free Mass(FFM)-For-age Z-score Primary · 36 weeks or hospital discharge, up to 120 days following birth, whichever is longer

Estimated by air displacement plethysmography. FFM measurements were converted into Z-scores using updated, sex-specific reference curves of body composition in preterm infants (Norris et al, 2019). A Z-score of 0 represents the population mean. Z-score values closer to 0 represent a better outcome. No relevant thresholds have been defined.

Group	Value	95% CI
Protein-enriched Human Milk Diet	-1.7	± 1.4
Usual Human Milk Diet	-1.6	± 1.5

Fat Mass(FM)-For-age Z-score Secondary · 36 weeks or hospital discharge, up to 120 days following birth, whichever is longer

Estimated by air displacement plethysmography. FM measurements were converted into Z-scores using updated, sex-specific reference curves of body composition in preterm infants (Norris et al, 2019). A Z-score of 0 represents the population mean. Z-score values closer to 0 represent a better outcome. No relevant thresholds have been defined.

Group	Value	95% CI
Protein-enriched Human Milk Diet	1.0	± 1.1
Usual Human Milk Diet	1.3	± 1.2

Body Fat(BF)-For-age Z-score Secondary · 36 weeks or hospital discharge, up to 120 days following birth, whichever is longer

Body fat estimated by air displacement plethysmography. BF measurements were converted into Z-scores using updated, sex-specific reference curves of body composition in preterm infants (Norris et al, 2019). A Z-score of 0 represents the population mean. Z-score values closer to 0 represent a better outcome. No relevant thresholds have been defined.

Group	Value	95% CI
Protein-enriched Human Milk Diet	1.8	± 1
Usual Human Milk Diet	1.9	± 1.2

Anthropometric Measurements Secondary · Birth to 36 weeks postmenstrual age or hospital discharge (whichever occurred first)

Weight, length, and head circumference measurements. Measurements were converted into Z-scores based on Fenton growth curves (2013). They were calculated at birth and at 36 weeks. This outcome represents the change in weight, length, and head circumference Z-score during the course of the study (i.e., the Z-score at birth was subtracted from the Z-score at 36 weeks). A value of 0 represents that the infant's Z-score is the same at the beginning and the end of the study. Positive values indicate the increase in the infant's Z-score during the study; negative values indicate the decrease in the

Weight

Group	Value	95% CI
Protein-enriched Human Milk Diet	-1.0	± 0.7
Usual Human Milk Diet	-1.2	± 0.8

Length

Group	Value	95% CI
Protein-enriched Human Milk Diet	-1.5	± 1.0
Usual Human Milk Diet	-1.9	± 1.3

Head circumference

Group	Value	95% CI
Protein-enriched Human Milk Diet	-0.9	± 0.8
Usual Human Milk Diet	-1.3	± 1.1

Growth Rate Secondary · Birth to 36 weeks postmenstrual age or hospital discharge (whichever occurred first)

Weight gain in g/kg/day

Group	Value	95% CI
Protein-enriched Human Milk Diet	14.7	± 2.5
Usual Human Milk Diet	13.9	± 2.5

Number of Participants With Postnatal Growth Failure Secondary · 36 weeks or hospital discharge (whichever occurred first)

Diagnosis of growth failure (weight \< 10th percentile using the 2013 Fenton growth curves)

Group	Value	95% CI
Protein-enriched Human Milk Diet	31
Usual Human Milk Diet	30

Number of Participants With Diagnosis of Necrotizing Enterocolitis Secondary · From birth up to 120 days following birth

Diagnosis of necrotizing enterocolitis stage 2 or 3

Group	Value	95% CI
Protein-enriched Human Milk Diet	1
Usual Human Milk Diet	3

Number of Participants With Diagnosis of Intestinal Perforation Secondary · From birth up to 120 days following birth

Diagnosis of intestinal perforation

Group	Value	95% CI
Protein-enriched Human Milk Diet	5
Usual Human Milk Diet	4

Death Secondary · Birth to 120 days

Death prior to 121 days of life

Group	Value	95% CI
Protein-enriched Human Milk Diet	6
Usual Human Milk Diet	5

Culture-proven Sepsis Secondary · Birth to 120 days

Diagnosis of sepsis with positive blood cultures

Group	Value	95% CI
Protein-enriched Human Milk Diet	22
Usual Human Milk Diet	16

Number of Days Alive and Receiving Full Enteral Feeding Secondary · Birth to 28 days

Time to full enteral feeding days

Group	Value	95% CI
Protein-enriched Human Milk Diet	18	11 – 21
Usual Human Milk Diet	14	5 – 18

Duration of Hospital Stay in Days Secondary · Birth to 120 days or discharge, whichever occurs first

From day of admission to day of hospital discharge to home

Group	Value	95% CI
Protein-enriched Human Milk Diet	107	87 – 120
Usual Human Milk Diet	107	79 – 120

Adverse events — posted to ClinicalTrials.gov

Time frame: From birth up to 120 days following birth. Reporting threshold: 0%. Adverse-event reports describe events observed during the trial — not all are caused by the drug.

Protein-enriched Human Milk Diet

Serious: 6/75 (8%)

Deaths: 6/75

Usual Human Milk Diet

Serious: 4/75 (5%)

Deaths: 5/75

Serious adverse events (2 terms)

Reaction	System	Protein-enriched Human Mil…	Usual Human Milk Diet
Spontaneous intestinal perforation	Gastrointestinal disorders	—	—
Necrotizing enterocolitis	Gastrointestinal disorders	—	—

Other adverse events (1 terms — click to expand)

Reaction	System	Protein-enriched Human Mil…	Usual Human Milk Diet
Growth failure	General disorders	—	—

Most-reported serious reactions: Spontaneous intestinal perforation, Necrotizing enterocolitis.

Data from ClinicalTrials.gov NCT04325308 adverse events section.

Sponsor's own description

The central hypothesis of this clinical trial is that, in extremely preterm infants, protein-enriched human milk diets compared to usual human milk diets during the first 2 weeks after birth increase fat-free mass (FFM)-for-age Z scores and promote maturation of the gut microbiome at term corrected age.

Publications & conference data

2 peer-reviewed publications reference this trial (live from Europe PMC):

1. Early Human Milk Fortification in Infants Born Extremely Preterm: A Randomized Trial.
   Salas AA, Gunawan E, Nguyen K, Reeves A, et al · · 2023 · cited 28× · PMID 37551512 · DOI 10.1542/peds.2023-061603
2. Necrotizing Enterocolitis: What's New and What's Next?
   Sha C, Sander WR, Bass K, Hsieh H, et al · · 2025 · cited 1× · PMID 41096926 · DOI 10.3390/ijms26199660