Use of slpi for treating chronic inflammatory intestinal diseases

• US 20040106564A1
• Filed: 01/05/2004
• Published: 06/03/2004
• Est. Priority Date: 01/17/2001
• Status: Abandoned Application

• Current Assignee: Vivotec Biomedical Technologies GmbH
• Original Assignee: Vivotec Biomedical Technologies GmbH
• Inventors: Nilius, Manfred
• Application Number:
  US10/250,901
• Publication Number:
  US 20040106564A1
• Time in Patent Office: Days
• US Class Current: 514/44
• CPC Class Codes: A61K 38/57 from animals; from humans A61K38/553, A61K38/556 take precedence
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: C07K 14/811 Serine protease (E.C. 3.4.21) inhibitors

Abstract

The present invention relates to the use of secretory leucocyte protease inhibitor (SLPI), or a non-pathogenic microorganism capable of forming SLPI and containing a nucleic acid coding for SLPI, for the treatment of chronic inflammatory intestinal diseases of humans and animals, pharmaceutical compositions for oral or rectal administration which contain the effective material SLPI or SLPI-expressing microorganisms, and methods for the production of these pharmaceutical compositions.

Claims

1. Use of a effective material selected from the group consisting of secretory leucocyte protease inhibitor (SLPI), a fragment thereof, a complex thereof, a derivative thereof, an analog thereof, an expressible nucleic acid coding for the effective material SLPI or a fragment or derivative thereof, and a non-pathogenic microorganism containing the nucleic acid and capable of SLPI formation, for the treatment of a disease of a human or animal body, selected from the group of chronic inflammatory intestinal diseases consisting of enteritis necroticans, enteritis regionalis Crohn (Crohn'"'"'s disease), colitis cystica, colitis granulomatosa, colitis gravis, colitis haemorrhagia, colitis ischaemica, colitis mucosa and colitis ulcerosa (ulcerative colitis).

2. Use according to claim 1, wherein the treatment takes place by the administration of the isolated and purified effective material in a pharmaceutical composition.

3. Use according to claim 2, wherein the effective material is administered in a dose which is sufficient to heal the chronic inflammatory intestinal disease state or to prevent it, to stop the progression of chronic inflammatory intestinal disease and/or to alleviate the chronic inflammatory intestinal disease symptoms.

4. Use according to claim 2 or 3, wherein the effective material is administered once through three times daily in a dose of 1-5,000 mg of effective material.

5. Use according to one of claims 2-4, wherein the effective material is administered orally.

6. Use according to claim 5, where in the effective material is administered in the form of a suspension, tablet, pill, capsule, lollipop, granulate or powder.

7. Use according to one of claims 2-4, wherein the effective material is administered rectally.

8. Use according to claim 7, wherein the effective material is administered in the form of a suppository, enema or foam.

9. Use according to one of claims 2-4, wherein the effective material is administered parenterally.

10. Use according to claim 9, wherein the effective material is administered in the form of an injection or infusion.

11. Use according to claim 1, wherein the non-pathogenic microorganism is capable of producing the effective material before, during or after administration to a human or animal and to release the produced effective material after administration to the organs of the digestive tract.

12. Use according to claim 11, wherein the non-pathogenic microorganism is a bacterial or fungal microorganism which belongs to the commensals of humans or animals.

13. Use according to claim 12, wherein the fungal microorganism belongs to the genus Saccharomyces.

14. Use according to claim 11, wherein the fungal microorganism is Saccharomyces boulardii.

15. Use according to claim 12, wherein the non-pathogenic microorganism belongs to the natural intestinal flora of humans or animals.

16. Use according to claim 15, wherein the non-pathogenic microorganism is an aerobic or anaerobic gram-negative bacterium of the intestinal flora.

17. Use according to claim 16, wherein the gram-negative bacterium belongs to the genus Escherichia, Pseudomonas, Bacteroides, or Proteus.

18. Use according to one of claims 17 [sic], wherein the gram-negative bacterium is Escherichia coli (Nissle 1917).

19. Use according to claim 15, wherein the non-pathogenic microorganism is an aerobic or anaerobic gram-positive bacterium of the intestinal flora.

20. Use according to claim 19, wherein the gram-positive bacterium belongs to the genus Bifidobacterium, Streptococcus, Staphylococcus, or Corynebacterium.

21. Use according to claim 20, wherein the gram-positive bacterium is Streptococcus gordonii.

22. Use according to claim 11, wherein the non-pathogenic microorganism is a microorganism which does not belong to the commensals of humans or animals.

23. Use according to claim 22, wherein the non-pathogenic microorganism is a bacterium which is used for the fermentative production of foodstuffs.

24. Use according to claim 22 or 23, wherein the bacterium concerned is a lactic acid bacterium, such as Lactococcus lactis, Lactobacillus delbrueckii subspec. bulgaricus, Lactobacillus casei, Lactobacillus caucasicus, Lactobacillus kefir, Streptococcus thermophilus, or Leconostoc.

25. Use according to one of claims 11-24, wherein a “

leaky”

mutant is concerned as the microorganism.

26. Use according to claim 11-25, wherein the nucleic acid coding for SLPI or a fragment or derivative thereof is inserted into a vector.

27. Use according to claim 26, wherein the vector is a plasmid, cosmid, bacteriophage or virus.

28. Use according to claim 26 or 27, wherein the nucleic acid inserted into a vector is under the functional control of at least one regulating element, which ensures the transcription of the nucleic acid in a translatable RNA and/or the translation of the RNA into a protein, before, during or after the administration.

29. Use according to claim 28, wherein the at least one regulating element is a promoter, a ribosome binding site, a signal sequence or a 3′

-transcription terminator.

30. Use according to claim 29, wherein the promoter is an inducible promoter.

31. Use according to claim 30, wherein the promoter is a promoter which is inducible by nutrient deficiency.

32. Use according to claim 30 or 31, wherein the promoter is a trp-, lac- or tac-promoter of Escherichia coli.

33. Use according to claim 30 or 31, where in the promoter is the phoA promoter of Escherichia coli,

34. Use according to claim 30 or 31, wherein the promoter is the promoter of the PHO 5 gene of yeast.

35. Use according to claim 30 or 31, wherein the promoter is the promoter of the ADH 1 gene of yeast.

36. Use according to one of claims 29-35, wherein the ribosome binding site is a Shine-Dalgarno sequence.

37. Use according to one of claims 29-36, wherein the signal sequence is a bacterial or fungal signal sequence, which effects the secretion of the protein out of the cytoplasm of the microorganism into the periplasmic space or into the environment of the microorganism.

38. Use according to claim 37, wherein, as the bacterial signal sequence, the signal sequence of the β

-lactamase gene of Escherichia coli or the signal sequence of the ompA gene of Escherichia coli is concerned.

39. Use according to claim 37, wherein, as the fungal signal sequence, the signal sequence of the α

-factor of yeast or the signal sequence of the killer toxin of yeast is concerned.

40. Use according to one of claims 11-39, wherein the non-pathogenic microorganism capable of SLPI formation is contained in a pharmaceutical composition.

41. Use according to claim 40, wherein the pharmaceutical composition containing the microorganism is administered orally.

42. Use according to claim 41, wherein the pharmaceutical composition containing the microorganism is administered in the form of a suspension, tablet, pill, capsule, granulate or powder.

43. Use according to claim 40, wherein the pharmaceutical composition containing the microorganism is administered rectally.

44. Use according to claim 43, wherein the pharmaceutical composition containing the microorganism is administered in the form of a suppository, enema, or foam.

45. Pharmaceutical composition, comprising at least one cell of a non-pathogenic microorganism capable of forming SLPI and containing an expressible nucleic acid coding for SLPI or a fragment or derivative thereof.

46. Pharmaceutical composition according to claim 45, w herein the microorganism is an anaerobic or aerobic, gram-negative or gram-positive, bacterium of the intestinal flora.

47. Pharmaceutical composition according to claim 45, wherein the microorganism is a commensal yeast of humans or animals.

48. Pharmaceutical composition according to claim 45, wherein the microorganism is a bacterium which can be used for the fermentative production of foodstuffs.

49. Pharmaceutical composition according to claim 45, wherein the microorganism is a “

leaky”

mutant.

50. Pharmaceutical composition according to one of claims 45-49, wherein a nucleic acid coding for SLPI or a fragment or derivative thereof is inserted into an expression vector, and wherein the expression of the nucleic acid is under the control of at least one regulating element, so that the effective material is expressed before, during or after the administration of the pharmaceutical composition, and is released to the organs of the digestive tract after the administration of the pharmaceutical composition.

51. Method of production of a pharmaceutical composition, comprising:

(a) isolation or synthesis of a nucleic acid coding for the effective material SLPI;

(b) cloning of the nucleic acid coding for SLPI in a bacterial expression vector or a fungal expression vector;

(c) transformation of the recombinant expression vector obtained in (b) in a microbial host cell, where the host cell is a commensal of the human or animal intestinal flora and/or can be used for the fermentative production of foodstuffs;

(d) propagation of the transformed host cells;

(e) production of an immobilized, lyophilized, or liquid preparation of transformed host cells;

(f) mixing the immobilized, lyophilized, preparation or suspension of transformed host cells obtained in (e) with physiologically acceptable excipients, stabilizers, thickeners, parting agents, lubricants, emulsifiers or the like materials to obtain a pharmaceutical composition.