# US 2023/0263178 A1 · Bovine milk exosome products and methods, nutritional compositions, and therapeutic methods

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- Number: US 2023/0263178 A1
- Document ID: US20230263178A1
- Status: Published application
- Kind: A1
- Application number: 17/907,750
- Filed: 2021-04-01
- Published: 2023-08-24
- Earliest claimed: 2020-04-03
- Assignees: Abbott Laboratories
- Applicants: Abbott Laboratories
- Inventors: José María LÓPEZ PEDROSA; Ricardo RUEDA CABRERA; Jorge GARCÍA MARTÍNEZ
- Web page: https://patentgrep.com/patent/US20230263178A1

## CPC

- A23C 9/1425: {by ultrafiltration, microfiltration or diafiltration of whey, e.g. treatment of the UF permeate}
- A23L 33/13: Nucleic acids or derivatives thereof (A23L33/145 takes precedence)
- A23C 9/1512: {containing isolated milk or whey proteins, caseinates or cheese; Enrichment of milk products with milk proteins in isolated or concentrated form, e.g. ultrafiltration retentate}
- A23C 9/20: Dietetic milk products not covered by groups A23C9/12 - A23C9/18
- A23L 33/40: {Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula}
- A61K 9/0095: {Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches (A61K9/0007 takes precedence; eatable gels or foams A61K9/0056; oral mucosa adhesive forms A61K9/006)}
- A61K 9/1276: Globules of milk; Constituents thereof
- A61K 35/12: Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells (vaccines or medicinal preparations containing antigens or antibodies A61K39/00)
- A61K 35/20: Milk; Whey; Colostrum
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A23C: DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF (preservation thereof A23B11/00)
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

A method of obtaining an exosome-enriched product comprises providing a whey-containing bovine milk fraction, conducting a first centrifugation of the whey-containing bovine milk fraction to obtain a whey middle fraction, conducting a second centrifugation of the whey middle fraction at an increased speed to obtain a concentrated whey fraction, filtering the concentrated whey fraction to obtain a filtered whey fraction, and conducting a third centrifugation of the filtered whey fraction at a further increased speed to obtain an exosome-enriched product. The exosome-enriched product comprises intact exosomes and less than 5 wt % casein based on the total weight of protein in the exosome-enriched product. Nutritional compositions comprise protein, carbohydrate, and/or fat, and exosomes provided by addition of the exosome-enriched product. A method of improving insulin sensitivity in a subject comprises administering a nutritional composition comprising the exosome-enriched product.

## Claims

### Claim 1

A method of obtaining an exosome-enriched product, the method comprising:
providing a whey-containing bovine milk fraction;
conducting a first centrifugation, comprising centrifuging the whey-containing bovine milk fraction to obtain a whey middle fraction;
conducting a second centrifugation, comprising centrifuging the whey middle fraction at an increased speed as compared with the speed of the first centrifugation to obtain a concentrated whey fraction;
filtering the concentrated whey fraction to obtain a filtered whey fraction; and
conducting a third centrifugation, comprising centrifuging the filtered whey fraction at an increased speed as compared with the speed of the second centrifugation to obtain an exosome-enriched product, wherein the exosome-enriched product comprises intact exosomes and less than 5 wt % casein based on the total weight of protein in the exosome-enriched product.

### Claim 2 (depends on 1)

The method of claim 1, wherein the whey-containing bovine milk fraction is provided by lowering the pH of a bovine milk product to precipitate milk solids, and removing the milk solids.

### Claim 3 (depends on 2)

The method of claim 2, wherein the pH of the milk product is lowered to about 3.0 to 4.6.

### Claim 4 (depends on 1)

The method of claim 1, wherein the whey-containing bovine milk fraction comprises.

### Claim 5 (depends on 4)

The method of claim 4, wherein the cheese whey is sweet cheese whey and has a pH from about 6.0 to about 6.5.

### Claim 6 (depends on 5)

The method of claim 5, wherein the sweet cheese whey comprises liquid separated from curd formed by subjecting bovine milk to at least one enzyme which destabilizes casein.

### Claim 7 (depends on 1)

The method of claim 1, wherein the second centrifugation comprises centrifuging the whey middle fraction at least two times to obtain the concentrated whey fraction.

### Claim 8 (depends on 1)

The method of claim 1, wherein the exosome-enriched product comprises at least 10 wt % exosomes.

### Claim 9 (depends on 8)

The method of claim 8, wherein at least about 50 wt % of exosomes in the exosome-enriched product are intact.

### Claim 10 (depends on 9)

The method of claim 9, wherein at least about 55, 60, 65, 70, 75, 80, 85, 90, or 95 wt % of the exosomes in the exosome-enriched product are intact.

### Claim 11 (depends on 1)

The method of claim 1, wherein the exosome-enriched product comprises less than 4, 3, 2, 1, 0.5, or 0.1 wt % casein based on the total weight of protein in the exosome-enriched product.

### Claim 12 (depends on 1)

The method of claim 1, wherein the third centrifugation comprises ultracentrifuging the filtered whey fraction to obtain a pellet containing intact exosomes.

### Claim 13 (depends on 12)

The method of claim 12, further comprising incubating the pellet containing exosomes in aqueous medium to dissolve the pellet without disrupting exosome membranes to provide an exosome suspension.

### Claim 14 (depends on 13)

The method of claim 13, further comprising drying the exosome suspension.

### Claim 15 (depends on 14)

The method of claim 14, wherein the drying comprises freeze drying the exosome suspension.

### Claim 16 (depends on 1)

The method of claim 1, wherein the exosome-enriched product is an exosome-enriched powder.

### Claim 17 (depends on 1)

An exosome-enriched product obtained by the method of claim 1.

### Claim 18

A bovine milk-derived exosome-enriched product comprising bovine milk-derived intact exosomes and less than 5 wt % casein based on the total weight of protein in the exosome-enriched product.

### Claim 19 (depends on 18)

The bovine milk-derived exosome-enriched product of claim 18, wherein the exosome-enriched product comprises less than 4, 3, 2, 1, 0.5, or 0.1 wt % casein based on the total weight of protein in the exosome-enriched product.

### Claim 20 (depends on 19)

The bovine milk-derived exosome-enriched product of claim 19, wherein the exosome-enriched product comprises at least 10 wt % exosomes.

### Claim 21 (depends on 20)

The bovine milk-derived exosome-enriched product of claim 20, wherein at least about 50 wt % of exosomes in the exosome-enriched product are intact.

### Claim 22 (depends on 21)

The bovine milk-derived exosome-enriched product of claim 21, wherein at least about 55, 60, 65, 70, 75, 80, 85, 90, or 95 wt % of the exosomes in the exosome-enriched product are intact.

### Claim 23 (depends on 22)

The bovine milk-derived exosome-enriched product of claim 22, wherein the exosome-enriched product is a powder.

### Claim 24 (depends on 18)

The bovine milk-derived exosome-enriched product of claim 18, wherein the exosomes are included in a nutritional composition comprising protein, carbohydrate, and/or fat, and the exosomes.

### Claim 25 (depends on 24)

The nutritional composition of claim 24, comprising from about 0.001 to about 10 wt % of the bovine milk-derived exosomes, based on the weight of the nutritional composition.

### Claim 26 (depends on 24)

The nutritional composition of claim 24, wherein the protein comprises whey protein concentrate, whey protein isolate, whey protein hydrolysate, milk protein concentrate, milk protein isolate, milk protein hydrolysate, soy protein concentrate, soy protein isolate, soy protein hydrolysate, pea protein concentrate, pea protein isolate, pea protein hydrolysate, fava bean protein, or combinations of two or more thereof.

### Claim 27 (depends on 24)

The nutritional composition of claim 24, wherein the carbohydrate comprises fiber, human milk oligosaccharides (HMOs), maltodextrin, corn maltodextrin, corn syrup, sucralose, cellulose gel, cellulose gum, gellan gum, carrageenan, fructooligosaccharides, or combinations of two or more thereof.

### Claim 28 (depends on 24)

The nutritional composition of any one of claim 24, wherein the fat comprises coconut oil, fractionated coconut oil, soy oil, soy lecithin, corn oil, safflower oil sunflower oil, palm olein, canola oil monoglycerides, lecithin, at least one omega-3 fatty acid, anola oil, medium chain triglycerides, one or more fatty acids such as linoleic acid, alpha-linolenic acid, ARA, EPA, and/or DHA, or combinations of two or more thereof.

### Claim 29 (depends on 28)

The nutritional composition of claim 28, wherein the at least one omega-3 fatty acid of the composition is selected from the group consisting of eicosapentaenoic acid, docosahexaenoic acid, arachidonic acid, and alpha-linolenic acid.

### Claim 30 (depends on 29)

The nutritional composition of claim 29, wherein the nutritional composition is in the form of a powder.

### Claim 31 (depends on 29)

The nutritional composition of claim 29, wherein the nutritional composition is in the form of a liquid.

### Claim 32 (depends on 18)

A method of improving insulin sensitivity in an individual at risk of developing or having insulin-resistance, prediabetes, or diabetes, comprising:
administering to the individual a nutritional composition comprising the exosome-enriched product of claim 18.