# US 11,299,520 B2 · Compositions comprising a chimeric bacterial surface layer protein and methods of use thereof

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- Number: US 11,299,520 B2
- Document ID: US11299520B2
- Status: Granted
- Kind: B2
- Application number: 16/651,160
- Filed: 2018-09-25
- Published: 2020-07-30
- Granted: 2022-04-12
- Earliest claimed: 2017-09-26
- Assignees: Arizona Board of Regents on Behalf of the University of Arizona
- Applicants: Arizona Board of Regents on Behalf of the University of Arizona
- Inventors: Gayatri Vedantam; Virinchipuram Viswanathan
- Web page: https://patentgrep.com/patent/US11299520B2

## CPC

- A61K 39/08: Clostridium, e.g. Clostridium tetani
- C07K 14/33: from Clostridium (G)
- A61K 39/09: {Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus}, streptococcus
- A61P 31/04: Antibacterial agents
- C07K 14/335: from Lactobacillus (G)
- C12N 9/90: Isomerases (5.)
- C12N 15/62: DNA sequences coding for fusion proteins
- A61K 9/0053: {Mouth and digestive tract, i.e. intraoral and peroral administration}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- A61K 2039/52: {Bacterial cells; Fungal cells; Protozoal cells}
- A61K 2039/523: {expressing foreign proteins}
- 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)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- 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)

## Abstract

The invention features engineered probiotic lacto acid bacteria (LAB) expressing a chimeric Clostridium difficile/Lactobacillus acidophilus SlpA, or fragment thereof, and its use for the treatment or prevention of Clostridium difficile infection and gut colonization.

## Claims

### Claim 1

A chimeric nucleic acid molecule comprising a nucleic acid sequence comprising a phosphoglycerate mutase (pgm) constitutive promoter (pgm promoter) operatively linked to a nucleic acid sequence encoding a chimeric bacterial surface layer protein A (SlpA) comprising a Clostridium difficile variable domain having at least 85% identity to a Clostridium difficile variable domain having the sequence corresponding to SEQ ID NO:15 and a lactic acid bacterium (LAB) peptidoglycan anchor having at least 85% identity to a LAB peptidoglycan anchor having the sequence corresponding to SEQ ID NO:14.

### Claim 2 (depends on 1)

The chimeric nucleic acid molecule of claim 1, wherein the pgm is from a Lactobacillus species selected from the group consisting of Lactobacillus acidophilus and Lactobacillus casei.

### Claim 3 (depends on 1)

The chimeric nucleic acid molecule of claim 1, wherein the LAB is Lactobacillus species selected from the group consisting of Lactobacillus acidophilus and Lactobacillus casei.

### Claim 4 (depends on 1)

The chimeric nucleic acid molecule of claim 1, wherein the nucleic acid sequence comprising the pgm promoter and the nucleic acid sequence encoding the chimeric bacterial SlpA has the nucleic acid sequence of SEQ ID NO: 18.

### Claim 5 (depends on 1)

A method of treating or preventing Clostridium difficile infection in a mammal, the method comprising administering to the mammal an engineered cell comprising the chimeric nucleic acid molecule of claim 1, thereby treating or preventing Clostridium difficile infection in the mammal.

### Claim 6 (depends on 5)

The method of claim 5, wherein the administered engineered cell is selected from the group consisting of Lactobacillus acidophilus, and Lactobacillus casei.

### Claim 7 (depends on 5)

The method of claim 5, wherein the engineered cell is administered orally.

### Claim 8 (depends on 5)

The method of claim 5, wherein the engineered cell is administered daily.

### Claim 9 (depends on 5)

The method of claim 5, wherein the mammal is a human.