# US 11,304,985 B2 · Compositions and methods for promoting a healthy microbial flora in a mammal

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- Number: US 11,304,985 B2
- Document ID: US11304985B2
- Status: Granted
- Kind: B2
- Application number: 16/468,053
- Filed: 2018-03-09
- Published: 2019-12-19
- Granted: 2022-04-19
- Earliest claimed: 2017-03-10
- Assignees: BIOHM Health LLC
- Applicants: BIOHM Health LLC
- Inventors: Afif Ghannoum
- Web page: https://patentgrep.com/patent/US11304985B2

## CPC

- A61K 35/741: Probiotics (probiotic yeast, e.g. saccharomyces A61K36/06)
- A61K 36/064: Saccharomycetales, e.g. baker's yeast
- A23L 31/00: Edible extracts or preparations of fungi; Preparation or treatment thereof
- A23L 33/135: Bacteria or derivatives thereof, e.g. probiotics
- A23L 33/14: Yeasts or derivatives thereof
- A23L 33/18: Peptides; Protein hydrolysates
- A61K 9/0053: {Mouth and digestive tract, i.e. intraoral and peroral administration}
- A61K 9/0056: {Mouth soluble or dispersible forms; Suckable, eatable, chewable coherent forms; Forms rapidly disintegrating in the mouth; Lozenges; Lollipops; Bite capsules; Baked products; Baits or other oral forms for animals}
- A61K 9/19: lyophilised {, i.e. freeze-dried, solutions or dispersions (lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00)}
- A61K 9/4808: {characterised by the form of the capsule or the structure of the filling; Capsules containing small tablets; Capsules with outer layer for immediate drug release (capsules filled with granules or microparticles A61K9/16; filled with microcapsules or coated microparticles A61K9/50; with mixture of different granules, microcapsules, (coated) microparticles A61K9/5084)}
- A61K 9/5047: {Cellulose ethers containing no ester groups, e.g. hydroxypropyl methylcellulose}
- A61K 35/745: Bifidobacteria
- A61K 35/747: Lactobacilli, e.g. L. acidophilus or L. brevis
- A61K 38/43: Enzymes; Proenzymes; Derivatives thereof
- A61K 38/47: acting on glycosyl compounds (3.2), e.g. cellulases, lactases
- A61P 31/04: Antibacterial agents
- A61P 31/10: Antimycotics
- C12N 1/16: Yeasts; Culture media therefor
- C12N 1/20: Bacteria; Culture media therefor
- C12Y 302/01001: Alpha-amylase (3.2.1.1)
- A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs
- A23V 2400/113: Acidophilus
- A23V 2400/175: Rhamnosus
- A23V 2400/519: Breve
- A61K 2035/115: {Probiotics}
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- 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)
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The disclosure provides compositions and dosage forms for promoting healthy microbial flora in a mammal, for example, a human. The disclosure further provides compositions and dosage forms for disrupting biofilm or preventing formation of biofilm containing pathogenic bacteria and/or pathogenic fungi in a particular region of a subject. The dosage forms contain (i) an isolated non-pathogenic fungal strain that is viable in the region of the subject, (ii) an isolated non-pathogenic bacterial strain that is viable in the region of the subject, and optionally (iii) an enzyme capable of disrupting the biofilm. The present disclosure also provides methods of disrupting a biofilm or preventing formation of a biofilm with such a composition or dosage form, methods of identifying a subject suitable for treatment with such a composition or dosage form, and methods of improving nutrient absorption using such a composition or dosage form.

## Claims

### Claim 1

A biofilm disrupting composition comprising (i) a freeze dried or spray dried blend comprising an isolated and viable non-pathogenic fungal strain and three different isolated and viable non-pathogenic bacterial strains and (ii) an amylase enzyme capable of disrupting a biofilm in a subject, wherein the isolated and viable non-pathogenic fungal strain is Saccharomyces boulardii and the three isolated and viable non-pathogenic bacterial strains are Lactobacillus rhamnosus, Bifidobacterium breve, and Lactobacillus acidophilus.

### Claim 2

A dosage form capable of disrupting a biofilm comprising a pathogenic bacteria and a pathogenic fungus disposed within a preselected region of a subject, the dosage form comprising a composition comprising (i) an amylase enzyme capable of disrupting the biofilm and (ii) a freeze dried or spray dried blend comprising a non-pathogenic fungal strain capable of replicating in the region of the subject and three different isolated and viable non-pathogenic bacterial strains capable of replicating in the region of the subject, wherein the isolated and viable non-pathogenic fungal strain is Saccharomyces boulardii and the three isolated and viable non-pathogenic bacterial strains are Lactobacillus rhamnosus, Bifidobacterium breve, and Lactobacillus acidophilus.

### Claim 3 (depends on 1)

The composition of claim 1, wherein the non-pathogenic bacterial strains of Lactobacillus rhamnosus, Bifidobacterium breve, and Lactobacillus acidophilus are: Lactobacillus rhamnosus LB 20, Bifidobacterium breve S 46, and Lactobacillus acidophilus RP 32, respectively.

### Claim 4 (depends on 3)

The composition of claim 3, wherein the non-pathogenic fungal strain of Saccharomyces boulardii is Saccharomyces boulardii SB 48.

### Claim 5 (depends on 1)

The composition of claim 1, wherein the amylase is selected from the group consisting of Bacillus stearothermophilus amylase, Bacillus amyloliquefaciens amylase, Bacillus subtilis amylase, Bacillus licheniformi amylase, Aspergillus niger amylase, and Aspergillus oryzae amylase.

### Claim 6 (depends on 5)

The composition of claim 5, wherein the composition comprises from about 100 to about 5,000 SKB units of amylase, from about 200 to about 4,000 SKB units of amylase, from about 300 to about 2,000 SKB units of amylase, or from about 400 to about 1,000 SKB units of amylase.

### Claim 7 (depends on 2)

The dosage form of claim 2, wherein the composition is disposed in a capsule.

### Claim 8 (depends on 2)

The dosage form of claim 2, wherein the composition is formulated as a granulate, pellet, or coated powder.

### Claim 9 (depends on 8)

The dosage form of claim 8, wherein the coated powder is coated with hydroxypropyl methylcellulose.

### Claim 10 (depends on 7)

The dosage form of claim 7, wherein the capsule comprises from about 50 mg to about 1,000 mg of the composition.

### Claim 11 (depends on 1)

The composition of claim 1, wherein the composition comprises from about 10 billion to about 40 billion, from about 15 billion to about 40 billion, from about 20 billion to about 40 billion, from about 10 billion to about 30 billion, from about 15 billion to about 30 billion, or from about 20 billion to about 30 billion colony forming units of Lactobacillus rhamnosus, Bifidobacterium breve, and Lactobacillus acidophilus.

### Claim 12 (depends on 11)

The composition of claim 11, wherein the composition comprises from about 1 billion to about 10 billion, from about 2 billion to about 8 billion, or from about 3 billion to about 6 billion colony forming units of Saccharomyces boulardii.

### Claim 13 (depends on 12)

The composition of claim 12, wherein the composition comprises about 15 billion colony forming units of Bifidobacterium breve, about 10 billion colony forming units of Lactobacillus rhamnosus, about 3.5 billion colony forming units of Saccharomyces boulardii, and about 1.5 billion colony forming units of Lactobacillus acidophilus.

### Claim 14 (depends on 1)

A method of disrupting a biofilm or preventing formation of a biofilm comprising a pathogenic bacteria and/or a pathogenic fungus disposed within a preselected region of a subject in need thereof, the method comprising administering to the subject the composition of claim 1, whereupon administration of the composition disrupts or prevents formation of said biofilm in the subject.

### Claim 15 (depends on 14)

The method of claim 14, wherein the pathogenic fungus is Candida tropicalis, Candida albicans, or a combination thereof.

### Claim 16 (depends on 14)

The method of claim 14, wherein the biofilm comprises (i) Candida tropicalis and Escherichia coli, (ii) Candida tropicalis and Serratia marcescens, or (iii) Candida tropicalis, Escherichia coli, and Serratia marcescens.

### Claim 17 (depends on 14)

The method of claim 14, wherein the biofilm comprises E. coli and a Bacteroides spp., wherein the Bacteroides spp. is selected from the group comprising Bacteroides coprophilus, Bacteroides eggerthii, Bacteroides ovatus, Bacteroides fragilis, Bacteroides plebeius, and Bacteroides uniformis.

### Claim 18 (depends on 14)

The method of claim 14, wherein the biofilm is disposed in the gastrointestinal tract, urinary tract, reproductive tract, upper respiratory tract, lower respiratory tract, biliary tract, mouth, eye, nose, ear, or skin.

### Claim 19 (depends on 14)

The method of claim 14, whereupon administration of the composition treats hyperammonemia, Clostridium difficile colitis, hepatic encephalopathy associated with cirrhosis, inflammatory bowel disease, Crohn's disease, irritable bowel disease, or any combination(s) thereof.

### Claim 20 (depends on 14)

The method of claim 14, whereupon administration of the composition permits restoration of the natural microbiome of the region in the subject.