Microbiome-based informed method to formulate live biotherapeutics

Patent metadata

Field	Value
Publication number	US-11389491-B2
Application number	US-202117223710-A
Country	US
Kind code	B2
Filing date	Apr 6, 2021
Priority date	Oct 16, 2019
Publication date	Jul 19, 2022
Grant date	Jul 19, 2022

Abstract

Methods of formulating live biotherapeutics are disclosed in which a deficiency or excess of a specific bacterial strain in a person's microbiome is identified by comparing a gene-specific characterization of the person's microbiome against a comprehensive, non-redundant reference gene catalog, and the biotherapeutic is formulated by selecting bacteria to address the deficiency or excess. Embodiments include the formulation of live biotherapeutics for improving the health of a person's vaginal microbiome, i.e. using a vaginal reference gene catalog, and may be suitable for ameliorating, treating, or preventing a malignancy such as a cancer of the female genitourinary system.

First claim

The invention claimed is:

1. A method for achieving an improvement in the vaginal health of a female human subject, comprising:
administering a remedial live biotherapeutic formulation to the subject,
wherein the remedial live biotherapeutic formulation comprises bacteria adapted to remedy wherein the remedial live biotherapeutic formulation comprises bacteria adapted to remedy a deficiency or excess of at least one bacterial strain in the subject's vaginal microbial community,
wherein the bacteria comprise a selected strain or consortium of strains comprising at least one of:
(i) Lactobacillus crispatus bacteria configured to express at least one of the asparagine synthase B (asnB) gene of SEQ ID NO: 2 and the asparagine synthase B (asnB) gene of SEQ ID NO: 3; and
(ii) Lactobacillus crispatus bacteria containing an asparagine synthase B (asnb) gene encoding at least one of a polypeptide of SEQ ID NO: 5 and a polypeptide of SEQ ID NO: 6, and
wherein the improvement is selected from the group consisting of decreasing ammonia in the vaginal environment of the subject, reducing inflammation in the subject, suppressing the growth or hampering the survival of at least one of Gardnerella vaginalis and Prevotella bivia in the vagina of the subject, and combinations thereof.

2. The method of claim 1 , wherein the selected strain or consortium of strains comprises at least one strain selected from the group consisting of LUCA111, LUCA011, LUCA015, LUCA009, LUCA102, LUCA006, LUCA059, LUCA103, and LUCA008.

3. The method of claim 1 , wherein the live biotherapeutic formulation further comprises a pharmaceutically acceptable carrier.

4. The method of claim 1 , wherein the live biotherapeutic formulation further comprises an agent adapted to reduce or remove free ammonia from the vaginal environment.

5. The method of claim 1 , wherein the Lactobacillus crispatus bacteria are configured to express the asparagine synthase B (asnB) gene of SEQ ID NO: 2.

6. The method of claim 1 , wherein the Lactobacillus crispatus bacteria are configured to express the asparagine synthase B (asnB) gene of SEQ ID NO: 3.

7. The method of claim 1 , wherein the Lactobacillus crispatus bacteria containing an asparagine synthase B (asnB) gene encode the polypeptide of SEQ ID NO: 5.

8. The method of claim 1 , wherein the Lactobacillus crispatus bacteria containing an asparagine synthase B (asnB) gene encode the polypeptide of SEQ ID NO: 6.

9. The method of claim 1 , wherein the live biotherapeutic formulation further comprises a redox potential control agent.

10. The method of claim 1 , wherein the live biotherapeutic formulation further comprises a pH buffer configured to maintain a healthy pH of the vaginal environment.

11. The method of claim 1 , wherein the live biotherapeutic formulation further comprises an antimicrobial agent.

12. The method of claim 1 , wherein the live biotherapeutic formulation further comprises a growth promoter.

13. A method for achieving an improvement in the vaginal health of a female human subject, comprising:
administering a remedial live biotherapeutic formulation to the subject,
wherein the remedial live biotherapeutic formulation comprises bacteria adapted to remedy a deficiency or excess of at least one bacterial strain in the subject's vaginal microbial community,
wherein the bacteria comprise a selected strain or consortium of strains comprising at least one of:
(i) Lactobacillus crispatus bacteria configured to express at least one of the asparagine synthase B (asnB) gene of SEQ ID NO: 2 and the asparagine synthase B (asnB) gene of SEQ ID NO: 3; and
(ii) Lactobacillus crispatus bacteria containing an asparagine synthase B (asnB) gene encoding at least one of a polypeptide of SEQ ID NO: 5 and a polypeptide of SEQ ID NO: 6, and

wherein the improvement comprises decreasing ammonia in the vaginal environment of the subject.

14. The method of claim 13 , wherein the live biotherapeutic formulation further comprises at least one of an antimicrobial agent, a redox potential control agent and a pH buffer configured to maintain a healthy pH of the vaginal environment.

15. The method of claim 13 , wherein the live biotherapeutic formulation further comprises a redox potential control agent.

16. The method of claim 13 , wherein the live biotherapeutic formulation further comprises a pH buffer configured to maintain a healthy pH of the vaginal environment.

17. A method for achieving an improvement in the vaginal health of a female human subject, comprising:
administering a remedial live biotherapeutic formulation to the subject,
wherein the remedial live biotherapeutic formulation comprises bacteria adapted to remedy a deficiency or excess of at least one bacterial strain in the subject's vaginal microbial community,
wherein the bacteria comprise a selected strain or consortium of strains comprising at least one of:
(i) Lactobacillus crispatus bacteria configured to express at least one of the asparagine synthase B (asnB) gene of SEQ ID NO: 2 and the asparagine synthase B (asnB) gene of SEQ ID NO: 3; and
(ii) Lactobacillus crispatus bacteria containing an asparagine synthase B (asnB) gene encoding at least one of a polypeptide of SEQ ID NO: 5 and a polypeptide of SEQ ID NO: 6, and

wherein the improvement comprises reducing inflammation in the subject.

18. The method of claim 17 , wherein the live biotherapeutic formulation further comprises at least one of an antimicrobial agent, a redox potential control agent and a pH buffer configured to maintain a healthy pH of the vaginal environment.

19. The method of claim 17 , wherein the live biotherapeutic formulation further comprises an antimicrobial agent.

20. The method of claim 17 , wherein the method further comprises suppressing the growth or hampering the survival of at least one of Gardnerella vaginalis and Prevotella bivia in the vagina of the subject.

Assignees

• Univ Maryland

Inventors

• Ravel Jacques
• France Michael
• Rutt Lindsay
• Ma Bing

Classifications

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• A61K35/74

  Bacteria (therapeutic use of a bacterial protein A61K38/00) · CPC title

• A61K35/742 Primary

  Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes · CPC title

• A61K35/747 Primary

  Lactobacilli, e.g. L. acidophilus or L. brevis · CPC title

• A61P15/02

  for disorders of the vagina · CPC title