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EP-4736670-A1 - BOVINE MILK-ISOLATED POWDERED EXOSOMES, NUTRITIONAL COMPOSITIONS AND METHODS

EP4736670A1EP 4736670 A1EP4736670 A1EP 4736670A1EP-4736670-A1

Abstract

Bovine milk-isolated powdered exosomes comprise intact exosomes. Nutritional compositions comprise protein, carbohydrate, and/or fat, and exosomes isolated from bovine milk. A method of preparing powdered exosomes comprises centrifuging bovine milk to form a lipid fraction top layer, a whey fraction middle layer, and a first pellet of cells and debris, separating the whey fraction and centrifuging the separated whey fraction to remove additional fat, casein aggregates and debris and form a substantially clear whey fraction, microfiltering the substantially clear whey fraction to remove residual debris, centrifuging the microfiltered whey fraction to obtain a pellet containing exosomes, incubating the exosomecontaining pellet in aqueous medium to dissolve the pellet without disrupting exosome membranes to provide an exosome suspension, and drying the suspension to obtain the powdered exosomes. Methods of lowering a risk of developing, or treating, insulin resistance, prediabetes, or diabetes in a subject employ exosomes isolated from bovine milk.

Inventors

  • LÓPEZ PEDROSA, José María
  • GARCÍA MARTÍNEZ, Jorge

Assignees

  • ABBOTT LABORATORIES

Dates

Publication Date
20260506
Application Date
20201105

Claims (16)

  1. Bovine milk-isolated powdered exosomes for use in lowering a risk of developing insulin resistance, prediabetes, or diabetes in a subject, or treating insulin resistance, prediabetes, or diabetes in a subject, the bovine milk-isolated powdered exosomes comprising intact exosomes.
  2. The bovine milk-isolated powdered exosomes for use of claim 1, wherein the subject is obese or has sarcopenia.
  3. A nutritional composition for use in lowering a risk of developing insulin resistance, prediabetes, or diabetes in a subject, or treating insulin resistance, prediabetes, or diabetes in a subject, the nutritional composition comprising protein, carbohydrate, and/or fat, and bovine milk-isolated powdered exosomes comprising intact exosomes.
  4. The nutritional composition for use of claim 3, wherein the nutritional composition comprises protein, carbohydrate, and fat.
  5. The nutritional composition for use of claim 3 or claim 4, wherein the nutritional composition comprises protein in an amount from about 1 wt% to about 30 wt% of the nutritional composition.
  6. The nutritional composition for use of any one of claims 3-5, wherein the nutritional composition comprises carbohydrate in an amount from about 5 wt% to about 70 wt% of the nutritional composition.
  7. The nutritional composition for use of any one of claims 3-6, wherein the nutritional composition comprises fat in an amount from about 1 wt% to about 30 wt% of the nutritional composition.
  8. The nutritional composition for use of any one of claims 3-7, wherein the nutritional composition comprises milk protein concentrate and/or soy protein isolate as proteins; canola oil, sunflower oil, medium chain triglycerides, and/or DHA as fats; and hydrolyzed corn starch, sucrose, and/or maltodextrin as carbohydrates.
  9. The nutritional composition for use of any one of claims 3-7, wherein the nutritional composition comprises milk protein concentrate as protein; corn maltodextrin, fructose, and/or glycerine as carbohydrates; and high oleic safflower oil as fat.
  10. The nutritional composition for use of any one of claims 3-9, wherein the nutritional composition has a caloric density of about 0.5 to about 3 kcal/mL.
  11. The nutritional composition for use of any one of claims 3-10, wherein the nutritional composition comprises β-hydroxy-β-methylbutyrate (HMB), preferably wherein the nutritional composition comprises HMB in an amount from about 0.01 wt% to about 10 wt% of the nutritional composition.
  12. The nutritional composition for use of any one of claims 3-11, wherein the composition is in powdered or liquid form.
  13. The bovine milk-isolated powdered exosomes for use of claim 1 or claim 2, or the nutritional composition for use of any one of claims 3-12, wherein the bovine milk-isolated powdered exosomes or the nutritional composition is for administration to the subject from 1 to 4 times per day.
  14. The bovine milk-isolated powdered exosomes for use of any one of claims 1, 2 or 13, or the nutritional composition for use of any one of claims 3-13, wherein administration to the subject increases an uptake of glucose by muscle cells of the subject.
  15. The bovine milk-isolated powdered exosomes for use of any one of claims 1, 2, 13, or 14, or the nutritional composition for use of any one of claims 3-14, wherein administration to the subject increases GLUT-4 transporter expression in a muscle of the subject.
  16. The bovine milk-isolated powdered exosomes for use of any one of claims 1, 2 or 13-15, or the nutritional composition for use of any one of claims 3-15, wherein the powdered exosomes or the nutritional composition is for use in enhancing muscle function in the subject, wherein the subject has insulin resistance.

Description

FIELD OF THE INVENTION The present invention relates to bovine milk-isolated powdered exosomes and to nutritional compositions containing protein, carbohydrate, and/or fat, and exosomes isolated from bovine milk. The present invention also relates to methods of preparing the bovine milk-isolated powdered exosomes, methods of preparing the nutritional compositions, and methods of lowering a risk of developing, or treating, insulin resistance, prediabetes, or diabetes in a subject by administering powdered exosomes or powdered nutritional compositions. BACKGROUND OF THE INVENTION Bovine milk contains exosomes which are extracellular bilayer vesicles. The bovine milk exosomes contain bioactive agents such as enzymatic and non-enzymatic proteins (CD9, CD63, MHC-class II, lactadherin, TSG101 and Hsc70, among others.), nucleic acids (including high amounts of microRNA (miRNA) and messenger RNA (mRNA)) and lipids (phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine and sphingomyelin, among others), that are known to have a role in regulating health and disease. Milk exosomes can be actively taken up by cells to deliver their content, but uptake requires the milk exosome bilayer membrane to be preserved. While the exosome structure protects bioactive agents, known methods of isolation of exosomes from milk can damage the exosome membrane, preventing uptake of the exosomes and resulting in release of bioactive agents and degradation of their bioactivity. For example, agents such as miRNA and other bioactive agents contained within the exosomes are extremely unstable and degrade within seconds of exposure when the bilayer membrane is damaged. Similarly, processing of milk and/or isolated exosomes, and/or extended storage of isolated exosomes, can damage the exosome membrane or otherwise degrade bioactivity. Accordingly, improved methods of isolating bovine milk exosomes and exosomes of improved stability are desirable. Insulin resistance, prediabetes, and diabetes are conditions of glucose dysregulation. Lifestyle modifications and insulin sensitizers are among the treatment strategies available, however these often fail and insulin therapy can become necessary. Dietary management can improve metabolic health and lead to weight loss. Planning for weight maintenance over time, sustaining and improving muscle mass, and taking vitamins and minerals can improve quality of life. Unfortunately, in many cases, lifestyle modifications are inadequate to fully treat such conditions. New compositions and methods for improving insulin sensitivity are therefore needed. SUMMARY OF THE INVENTION In one embodiment, the invention is directed to bovine milk-isolated powdered exosomes comprising intact exosomes. An intact exosome is one in which the bilayer membrane is undamaged and the contents of the exosome are retained within the exosome. In another embodiment, the invention is directed to a nutritional composition which comprises protein, carbohydrate, and/or fat, and exosomes isolated from bovine milk, wherein the exosomes are provided by addition of the powdered exosomes of the invention. In specific embodiments, the nutritional composition is in the form of a powder, and in additional specific embodiments, the nutritional composition is in the form of a liquid. In further embodiments, the nutritional composition comprises protein, carbohydrate, fat, and exosomes isolated from bovine milk, wherein the exosomes are provided by addition of the powdered exosomes of the invention. In an additional embodiment, the invention is directed to a method of preparing powdered exosomes. The method comprises centrifuging bovine milk to form a lipid fraction top layer, a whey fraction middle layer, and a first pellet of cells and debris, separating the whey fraction and centrifuging the separated whey fraction to remove additional fat, casein aggregates and debris and form a substantially clear whey fraction, microfiltering the substantially clear whey fraction to remove residual debris, centrifuging the microfiltered whey fraction to obtain a pellet containing exosomes, incubating the exosome-containing pellet in aqueous medium to dissolve the pellet without disrupting exosome bilayer membranes to provide an exosome suspension, and drying the suspension to obtain the powdered exosomes. In another embodiment, the invention is directed to a method of preparing a powdered nutritional composition. The method comprises dry blending a powder composition comprising protein, fat, and carbohydrate with powdered exosomes isolated from bovine milk. In further embodiments, the invention is directed to a method of lowering a risk of developing insulin resistance, prediabetes, or diabetes in a subject, or treating insulin resistance, prediabetes, or diabetes in a subject. The method comprises administering to a subject at risk of developing, or having, insulin resistance, prediabetes, or diabetes the exosomes according to the invention or the