Lactoferrin for Iron Regulation

Overseen By Abbie E. Smith-Ryan, PhD

Age: 18 - 65

Sex: Any

Trial Phase: Academic

Sponsor: University of North Carolina, Chapel Hill

Must not be taking: Antibiotics, Oral contraceptives

Disqualifiers: Anemia, Diabetes, Immunocompromised, others

Approved in 4 Jurisdictions This treatment is already approved in other countries

Compensation: Varies

Number of Visits: 8

110 Participants Needed

Trial Summary

What is the purpose of this trial?

Overall Objective: To determine the efficacy of rbLf supplementation in a healthy adult population, specifically with regard to iron regulation (primary), gut health (secondary), and immune function (secondary). Purpose: The purpose of this study is to determine the effects of rbLf on iron regulation, exercise performance, gut health, and immune function as compared to cow's milk derived bLf. Participants: To account for an approximate \~10% dropout rate (and rounding up to ensure equal number of enrolled participants per group), n=25 per group for males (N=50) and n=30 per group for females (N=60) will be enrolled for a total sample size of N=110. Procedures: Participation in this study will include 6 in-person visits over 14 weeks. Anemia will be checked and excluded at the first visit using a single finger prick, followed by a 10-20 min exercise test on a treadmill to evaluate exercise capacity. On visit 2 participants will perform three 3-15 minute exercise tests on a treadmill with rest in between each test in order to determine exercise performance. A finger prick will be performed before and after each run to determine lactate levels. After visits 2 participants will be randomized to their respective supplement group recombinant Bovine Lactoferrin supplement (rbLf) (Male and Fame), a bovine-milk derived Lactoferrin supplement (Female), or a placebo (Male) once daily for 4 weeks. Blood samples will be obtained prior to and after each supplement phase to determine change in iron levels and red blood cell capacity. After a 2-week washout participants will return to complete the same testing outcomes and will receive the subsequent supplement. At visit 6 a final blood sample will be collected.

Will I have to stop taking my current medications?

The trial protocol does not specify if you need to stop taking your current medications. However, if you are using oral contraceptives with iron supplementation or are currently on antibiotics, you may not be eligible to participate.

What data supports the effectiveness of the treatment Lactoferrin for Iron Regulation?

Research shows that lactoferrin, a protein found in milk, can help with iron uptake in the body and has antibacterial properties. Studies on both bovine and human lactoferrin indicate that they can be biologically active and may help in treating conditions like iron deficiency anemia by improving iron levels. ^{1 2 3 4 5}

Is lactoferrin safe for human use?

Research on recombinant human lactoferrin (rhLF) in animals, specifically rats, showed no harmful effects even at high doses over a 13-week period. This suggests that rhLF is generally safe, as no toxicologically significant changes were observed. ^{6 7 8 9 10}

How is the drug Lactoferrin unique in treating iron regulation?

Lactoferrin is unique because it is an iron-binding protein that not only helps regulate iron absorption but also has additional benefits like antibacterial, antiviral, and immune-boosting effects. Unlike other treatments, it can act as a bacteriostatic agent by withholding iron from bacteria, which is essential for their growth. ^{1 11 12 13 14}

Research Team

Abbie Smith-Ryan, PhD

Principal Investigator

University of North Carolina, Chapel Hill

Eligibility Criteria

This trial is for healthy adults interested in how rbLf affects iron regulation, gut health, and immune function. Participants must be willing to undergo blood tests, take supplements daily for 4 weeks, and complete treadmill exercise tests. Those with anemia or unable to commit to the study schedule are excluded.

Inclusion Criteria

Body mass index less than 35 kg/m^2

Active: meeting American College of Sports Medicine exercise guidelines or exercising more than 2 days per week

I am between 18 and 42 years old.

Exclusion Criteria

I have a diagnosis of Irritable Bowel Disease, Crohn's, or Celiac.

I have chronic eczema or asthma.

Vegan (due to supplement ingredients)

Timeline

Screening

Participants are screened for eligibility to participate in the trial

1 week

1 visit (in-person)

Run-in

Participants undergo a 2-week run-in period before starting treatment

2 weeks

1 visit (in-person)

Treatment Phase 1

Participants receive their first assigned supplement (rbLf, cmdLf, or placebo) for 4 weeks

4 weeks

2 visits (in-person)

Washout

Participants undergo a 2-week washout period between treatments

2 weeks

1 visit (in-person)

Treatment Phase 2

Participants receive their second assigned supplement (rbLf, cmdLf, or placebo) for 4 weeks

4 weeks

2 visits (in-person)

Follow-up

Participants are monitored for safety and effectiveness after treatment

1 week

1 visit (in-person)

Treatment Details

Interventions

• Recombinant Bovine Lactoferrin

Trial Overview The study compares the effects of a recombinant Bovine Lactoferrin supplement (rbLf), cow's milk-derived Lactoferrin (bLf), and a placebo on iron levels and red blood cell capacity in men and women through regular supplementation followed by exercise testing over 14 weeks.

Participant Groups

Group I: Females: Recombinant Bovine Lactoferrin (rbLf), then Bovine milk derived Lactoferrin Experimental Treatment 2 Interventions

After a 2-week run-in period, this arm will begin treatment with recombinant bovine lactoferrin for 4 weeks and following a 2-week washout start bovine milk derived lactoferrin for 4 weeks.

Group II: Females: Bovine milk derived lactoferrin (cmdLf), then Recombinant bovine lactoferrin Active Control 2 Interventions

After a 2-week run-in period, this arm will begin treatment with bovine milk derived lactoferrin and then following a 2-week washout start recombinant bovine lactoferrin for 4 weeks.

Group III: Males: Recombinant Bovine Lactoferrin (rbLf), then placebo Active Control 2 Interventions

After a 2-week run-in period, this arm will begin treatment with recombinant bovine lactoferrin for 4 weeks and following a 2-week washout start placebo for 4 weeks.

Group IV: Males: Placebo, then recombinant bovine lactoferrin (rbLf) Placebo Group 2 Interventions

After a 2-week run-in period, this arm will begin treatment with placebo for 4 weeks and following a 2-week washout start recombinant bovine lactoferrin for 4 weeks.

Recombinant Bovine Lactoferrin is already approved in United States, European Union, Canada, Japan for the following indications:

Approved in United States as Lactoferrin for:

• Dietary supplement for iron regulation and immune function

Approved in European Union as Lactoferrin for:

• Dietary supplement for iron regulation and immune function

Approved in Canada as Lactoferrin for:

• Dietary supplement for iron regulation and immune function

Approved in Japan as Lactoferrin for:

• Dietary supplement for iron regulation and immune function

Who Is Running the Clinical Trial?

University of North Carolina, Chapel Hill

Lead Sponsor

Trials

1,588

Recruited

4,364,000+

TurtleTree Labs Inc

Collaborator

Trials

1

Recruited

110+

Findings from Research

Lactoferrin, an iron-binding glycoprotein found in milk and other fluids, plays a crucial role in iron uptake in the intestines and acts as a bacteriostatic agent by limiting iron availability to bacteria.

Beyond its iron-binding capabilities, lactoferrin is also involved in immune responses and may function as a growth factor and bactericidal agent, highlighting its diverse biological significance.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Lactoferrin: molecular structure and biological function. Lönnerdal, B., Iyer, S.[2022]

Commercially available bovine lactoferrin (CbLF) can mimic some of the beneficial effects of human lactoferrin (hLF) in infant formula, as it can resist digestion and bind to intestinal cells similarly to hLF.

CbLF promotes cell proliferation and differentiation, and enhances the expression of important immune factors, suggesting it could provide health benefits similar to those of breast milk when added to infant formula.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Bovine lactoferrin can be taken up by the human intestinal lactoferrin receptor and exert bioactivities. Lönnerdal, B., Jiang, R., Du, X.[2022]

Recombinant bovine lactoferrin (rbLf) was successfully produced with a yield of 15.3 mg/l and 90.3% purity, demonstrating its potential for further research in antimicrobial applications.

Both recombinant rbLf and its pepsin-digested peptides showed significant antibacterial activity, with effectiveness rates of 79.8% and 86.9%, respectively, indicating their potential as immunomodulators.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Expression and characterization of recombinant bovine lactoferrin in E. coli. García-Montoya, I., González-Chávez, SA., Salazar-Martínez, J., et al.[2013]

The study successfully produced recombinant human lactoferrin (rhLf) using a recombinant virus in BmN cells, resulting in a biologically active protein with a molecular mass of approximately 78 kDa.

This method allows for efficient production of rhLf, which could have various applications in medicine and nutrition due to its multifunctional properties.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Recombinant functional human lactoferrin expressed in baculovirus system. Liu, T., Zhang, YZ., Wu, XF.[2019]

In a study involving SD rats with iron deficiency anemia, treatment with iron saturated recombinant human lactoferrin (Fe-rhLf) significantly improved hemoglobin levels and red blood cell counts compared to the control group, indicating its efficacy in treating this condition.

The high dose of Fe-rhLf not only enhanced hematological parameters but also altered liver hepcidin and ferroportin expression, suggesting a mechanism of action that may involve regulation of iron metabolism.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

[Efficacy of iron saturated recombinant human lactoferrin on alleviating iron deficiency anemia in rats]. Fang, YH., Feng, X., Xu, H., et al.[2019]

In a study involving 100 weanling SD rats over 28 days, recombinant human lactoferrin (rhLF) showed no significant differences in food safety indicators compared to bovine lactoferrin (bLF), indicating that both are safe for consumption.

Both rhLF and bLF groups demonstrated improved serum iron levels and transferrin saturation compared to the control group, suggesting that these lactoferrins can enhance iron nutrition without adverse effects.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

[Animal experiment of recombinant human lactoferrin based on the 28 days repeated oral toxicity]. Ma, B., Yuan, X., Wang, X., et al.[2019]

Recombinant human lactoferrin (rhLF) was successfully produced in the milk of transgenic cattle at concentrations of grams per liter, demonstrating a viable method for large-scale biopharmaceutical production.

Both natural human lactoferrin and rhLF exhibited similar iron-binding properties and effectiveness in fighting infections in various mouse models, indicating that rhLF can serve as a functional substitute for natural hLF.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Large scale production of recombinant human lactoferrin in the milk of transgenic cows. van Berkel, PH., Welling, MM., Geerts, M., et al.[2013]

A study on the oral toxicity of recombinant human lactoferrin (rhLF) in Wistar rats showed no significant toxic effects after 13 weeks of daily administration at doses up to 2000 mg/kg body weight.

The findings suggest that the no-observed-adverse-effect level (NOAEL) for rhLF is at least 2000 mg/kg/day, indicating a strong safety profile for this substance when consumed orally.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Sub-chronic (13-week) oral toxicity study in rats with recombinant human lactoferrin produced in the milk of transgenic cows. Appel, MJ., van Veen, HA., Vietsch, H., et al.[2011]

Bovine Lactoferrin (bLf) at a low concentration (0.8 μM) significantly reduces biofilm production in Listeria isolates, which is important for preventing foodborne illnesses.

bLf enhances the effectiveness of antibiotics like oxytetracycline and streptomycin against E. coli, and ampicillin against Listeria, indicating its potential as a synergistic agent in combating antibiotic-resistant bacteria.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Bovine Lactoferrin can Decrease the In Vitro Biofilm Production and Show Synergy with Antibiotics Against Listeria and Escherichia coli Isolates. García-Borjas, KA., Ceballos-Olvera, I., Luna-Castro, S., et al.[2021]

Dietary lactoferrin, now available from bovine sources and recombinant technology, has shown potential antibacterial and antiviral effects in the intestine, possibly enhancing immune function through a unique receptor-mediated pathway.

Lactoferrin may improve iron status in infants and pregnant women, stimulate intestinal cell growth, and even inhibit cancer development, suggesting it could be beneficial for infection prevention, inflammation reduction, and overall health in vulnerable populations.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Nutritional roles of lactoferrin. Lönnerdal, B.[2019]

Both human and bovine lactoferrins show strong interactions with specific lectins, indicating their potential roles in immune response and microbial recognition due to their glycosylation patterns.

The study highlights that while both types of lactoferrins can bind to various lectins, their binding properties differ, suggesting that human lactoferrins may have unique bioactive sites that could influence their therapeutic applications.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Recognition roles of the carbohydrate glycotopes of human and bovine lactoferrins in lectin-N-glycan interactions. Yen, MH., Wu, AM., Yang, Z., et al.[2016]

Bovine lactoferrin (bLF) significantly stimulates the proliferation of human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner, indicating its potential as an immunomodulatory agent.

Only colostral bLF was found to induce the production of proinflammatory cytokines TNF-α and IL-6, particularly at a concentration of 50 µg/mL, suggesting that the source and iron content of bLF can influence its immunological effects.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Bovine lactoferrin enhances proliferation of human peripheral blood lymphocytes and induces cytokine production in whole blood cultures. Zaczyńska, E., Kocięba, M., Śliwińska, E., et al.[2018]

Lactoferrin is a powerful glycoprotein that plays a crucial role in the body's defense against infections and inflammation, primarily through its ability to bind iron and interact with various microbial components and cell receptors.

Research into lactoferrin's structure and function is ongoing, with potential applications in healthcare, including the development of recombinant human lactoferrin for therapeutic use.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Structure and biological actions of lactoferrin. Nuijens, JH., van Berkel, PH., Schanbacher, FL.[2019]

Lactoferrin is a multifunctional glycoprotein with antibacterial, antiviral, anticancer, and immunomodulatory properties, making it a promising candidate for various clinical applications.

Advancements in genetic engineering have enabled the high-yield production of recombinant lactoferrin, which retains its biological activity and could enhance its use in medical treatments.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Research progress in physicochemical characteristics of lactoferrin and its recombinant expression systems. Pang, XN., Hong, X., Wei, X., et al.[2017]

References

1. United States pubmed.ncbi.nlm.nih.gov

Lactoferrin: molecular structure and biological function. [2022]

2. United States pubmed.ncbi.nlm.nih.gov

Bovine lactoferrin can be taken up by the human intestinal lactoferrin receptor and exert bioactivities. [2022]

3. Netherlands pubmed.ncbi.nlm.nih.gov

Expression and characterization of recombinant bovine lactoferrin in E. coli. [2013]

4. China pubmed.ncbi.nlm.nih.gov

Recombinant functional human lactoferrin expressed in baculovirus system. [2019]

5. China pubmed.ncbi.nlm.nih.gov

[Efficacy of iron saturated recombinant human lactoferrin on alleviating iron deficiency anemia in rats]. [2019]

6. China pubmed.ncbi.nlm.nih.gov

[Animal experiment of recombinant human lactoferrin based on the 28 days repeated oral toxicity]. [2019]

7. United States pubmed.ncbi.nlm.nih.gov

Large scale production of recombinant human lactoferrin in the milk of transgenic cows. [2013]

8. England pubmed.ncbi.nlm.nih.gov

Sub-chronic (13-week) oral toxicity study in rats with recombinant human lactoferrin produced in the milk of transgenic cows. [2011]

9. Netherlands pubmed.ncbi.nlm.nih.gov

Bovine Lactoferrin can Decrease the In Vitro Biofilm Production and Show Synergy with Antibiotics Against Listeria and Escherichia coli Isolates. [2021]

10. England pubmed.ncbi.nlm.nih.gov

Nutritional roles of lactoferrin. [2019]

11. Netherlands pubmed.ncbi.nlm.nih.gov

Recognition roles of the carbohydrate glycotopes of human and bovine lactoferrins in lectin-N-glycan interactions. [2016]

12. Poland pubmed.ncbi.nlm.nih.gov

Bovine lactoferrin enhances proliferation of human peripheral blood lymphocytes and induces cytokine production in whole blood cultures. [2018]

13. United States pubmed.ncbi.nlm.nih.gov

Structure and biological actions of lactoferrin. [2019]

14. China pubmed.ncbi.nlm.nih.gov

Research progress in physicochemical characteristics of lactoferrin and its recombinant expression systems. [2017]