Vaginal lactobacillus medicant

• US 6,372,209 B1
• Filed: 01/05/2000
• Issued: 04/16/2002
• Est. Priority Date: 04/11/1997
• Status: Expired due to Term

• Current Assignee: Osel, Inc.
• Original Assignee: GYNE Logix Incorporated
• Inventors: Chrisope, Gerald L.
• Primary Examiner(s): Park, Hankyel T.
• Assistant Examiner(s): Brown, Stacy S.
• Application Number:
  US09/478,185
• Time in Patent Office: 832 Days
• Field of Search: 424/93.45, 435/243, 435/252.1, 435/252.9
• US Class Current: 424/93.45
• CPC Class Codes: A61K 2039/52 Bacterial cells; Fungal cells; Protozoal cells
• CPC Class Codes: A61K 2039/541 Mucosal route
• CPC Class Codes: A61K 35/747 Lactobacilli, e.g. L. acidophilus or L. brevis
• CPC Class Codes: A61K 39/02 Bacterial antigens
• CPC Class Codes: A61K 39/09 Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/02 for disorders of the vagina
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 33/02 Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 11/10 the carrier being a carbohydrate
• CPC Class Codes: C12R 2001/225 Lactobacillus

Abstract

Disclosed are novel isolated strains of bacteria of the genus Lactobacillus which are useful in a vaginal medicant. Also disclosed are medicants containing such Lactobacilli, a novel preservation matrix for microorganisms, a method for preserving microbial cells within a medicant, and methods for preventing and treating vaginal and gastrointestinal infections.

Claims

1. A vaginal medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

2. The vaginal medicant of claim 1, wherein said strain has an ability to produce greater than 10 ppm of H₂O₂ under effective culture conditions.

3. The vaginal medicant of claim 1, wherein said strain sustains colonization of vaginal epithelial cells in vivo for greater than 1 month.

4. The vaginal medicant of claim 1, wherein said strain adheres to vaginal epithelial cells when said strain is in a metabolically inactive state.

5. The vaginal medicant of claim 1, wherein a single cell of said strain is from about 1 micron to about 2 microns in width and from about 2 microns to about 4 microns in length.

6. The vaginal medicant of claim 1, wherein said strain produces greater than 0.75 mg/100 ml of lactic acid under effective culture conditions.

7. The vaginal medicant of claim 1, wherein said matrix maintains greater than 10⁷ viable cells for a period of greater than 12 months in vitro.

8. The vaginal medicant of claim 1, wherein said matrix maintains greater than 10⁶ viable cells for a period of greater than 12 months at a temperature from about 4°

C. to about 6°

C. in vitro.

9. The vaginal medicant of claim 1, wherein said matrix maintains greater than 10⁶ viable cells for a period of greater than 12 months at about room temperature in vitro.

10. The vaginal medicant of claim 1, wherein said matrix has a pH of from about 5.0 to about 7.0.

11. The vaginal medicant of claim 1, wherein said strain is Lactobacillus crispatus.

12. The vaginal medicant of claim 1, wherein said strain is Lactobacillus jensenii.

13. A method to reduce the risk of infection in a female from a vaginal infection, comprising administering to a female a vaginal medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a potyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

14. The method of claim 13, wherein said vaginal infection is an infection selected from the group consisting of bacterial vaginosis, symptomatic yeast vaginitis, gonorrhea, chlamydia, trichomoniasis, human immunodeficiency virus infection, urinary tract infection and pelvic inflammatory disease.

15. A method to treat a vaginal infection, comprising administering to a female having a vaginal infection a vaginal medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

16. A method to reduce the risk of infection of a human by human immunodeficiency virus (HIV), comprising administering to a human a medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithlelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 Ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

17. The method of claim 16, wherein administration of said medicant reduces the risk of HIV infection of said human by greater than two-fold.

18. A method to prevent infection of a human by symptomatic yeast vaginitis, comprising administering to a human a medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

19. The method of claim 18, wherein said strain is Lactobacillus crispatus CTV-05.

20. A method to reduce the risk of infection of a human by a gastrointestinal infection, comprising administering to a human a medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

21. A method to reduce the occurrence of preterm birth, comprising administering to a pregnant female a medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

22. A method to enhance metabolism of estrogen in the vagina and bowel, comprising administering to a female a vaginal medicant comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

23. A gastrointestinal medicant, comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

24. A vaginal medicant comprising:

(a) a bacterial culture of at least two isolated strains of Lactobacillus having identifying characteristics comprising;

(i) a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%; and

(ii) an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions; and

(b) a preservation matrix, comprising a biologically active binding agent, an antioxidant, a polyol, a carbohydrate and a proteinaceous material;

wherein said matrix maintains greater than 10⁶ viable cells of said culture for a period of greater than 12 months in vitro.

25. The vaginal medicant of claim 24, wherein said medicant is used to treat at least two vaginal infections selected from the group consisting of bacterial vaginosis, symptomatic yeast vaginitis, gonorrhea, chlamydia, trichomoniasis, human immunodeficiency virus infection, urinary tract infection and pelvic inflammatory disease.

26. The vaginal medicant of claim 24, wherein said bacterial culture comprises a first strain of the genus Lactobacillus which is effective for a first vaginal infection and a second strain of the genus Lactobacillus which is effective for treating a second vaginal infection.

27. The vaginal medicant of claim 26, wherein said first strain is Lactobacillus crispatus CTV-05 and wherein said second strain is of the species Lactobacillus jensenii.

28. A vaginal medicant, comprising:

(a) a bacterial culture of an isolated strain of the genus Lactobacillus;

(b) a preservation matrix, comprising, about 14% gelatin, about 0.5%, sodium ascorbate, about 2.5% dextrose, about 1.5% skim milk and about 6% xylitol;

wherein said matrix maintains greater than 10⁶ cells of said culture for a period of greater than 12 months in vitro; and

wherein said matrix preserves desirable characteristics of said Lactobacillus, said characteristics selected from the group consisting of an ability to adhere to vaginal epithelial cells in a metabolically inactive state, an ability to produce greater than 0.5 ppm of H₂O₂ under effective culture conditions, and a percent vaginal epithelial cell (VEC) cohesion value of greater than 50%.

29. A vaginal medicant comprising a culture of Lactobacillus crispatus CTV-05 (ATCC Designation No. 202225).