Title:
METHODS AND COMPOSITIONS FOR ALTERING MUCUS SECRETION
Document Type and Number:
WIPO Patent Application WO/2000/050062
Kind Code:
A2
Abstract:
Methods and compounds for increasing or decreasing mucus secretion in subjects, and particularly mucus secretion in the airways, are described. Methods of screening compounds for the ability to increase or decrease mucus secretion are also described.
Inventors:
LI YUEHUA (US)
MARTIN LINDA D (US)
ADLER KENNETH B (US)
MARTIN LINDA D (US)
ADLER KENNETH B (US)
Application Number:
PCT/US2000/005050
Publication Date:
August 31, 2000
Filing Date:
February 24, 2000
Export Citation:
Assignee:
UNIV NORTH CAROLINA STATE (US)
LI YUEHUA (US)
MARTIN LINDA D (US)
ADLER KENNETH B (US)
LI YUEHUA (US)
MARTIN LINDA D (US)
ADLER KENNETH B (US)
International Classes:
A61K9/127; A61K9/72; A61K31/00; A61K31/235; A61K31/35; A61K31/47; A61K31/7088; A61K31/739; A61K38/00; A61K38/17; A61K38/19; A61K48/00; G01N33/50; A61P1/00; A61P11/00; A61P11/10; A61P43/00; C07H21/00; C07K14/47; C07K16/18; C12N5/07; C12N5/071; C12N15/09; C12Q1/02; G01N33/15; G01N33/53; G01N33/68; (IPC1-7): A61K38/17; A61K31/7088; A61K31/35; A61K31/00; A61K9/72; A61K9/127; C07H21/00; G01N33/50; A61K38/19; A61K31/739; A61P11/00; A61P1/00
Domestic Patent References:
| WO1993000353A1 | 1993-01-07 |
Foreign References:
| US5858784A | 1999-01-12 |
Other References:
DRAY-CHARIER NATHALIE ET AL: "Regulation of mucin secretion in human gallbladder epithelial cells: Predominant role of calcium and protein kinase C." GASTROENTEROLOGY, vol. 112, no. 3, 1997, pages 978-990, XP000946031 ISSN: 0016-5085
NAKAMURA MASATSUGU ET AL: "Mucin-like glycoprotein secretion is mediated by cyclic-AMP and protein kinase C signal transduction pathways in rat corneal epithelium." EXPERIMENTAL EYE RESEARCH, vol. 66, no. 5, May 1998 (1998-05), pages 513-519, XP000946070 ISSN: 0014-4835
THELEN M ET AL: "TUMOR NECROSIS FACTOR ALPHA MODIFIES AGONIST-DEPENDENT RESPONSES IN HUMAN NEUTROPHILS BY INDUCING THE SYNTHESIS AND MYRISTOYLATION OF A SPECIFIC PROTEIN KINASE C SUBSTRATE" PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES, vol. 87, no. 15, 1990, pages 5603-5607, XP002147379 1990 ISSN: 0027-8424
ADLER K B ET AL: "Effects of inflammatory mediators and drugs on mucus secretion and mucociliary function." RESEARCH IN IMMUNOLOGY, vol. 149, no. 3, March 1998 (1998-03), pages 245-248, XP000946047 Meeting on New Therapeutic Approaches for Allergic Diseases of the Respiratory Tract;Paris, France; April 1-4, 1998 ISSN: 0923-2494
ADLER KENNETH B ET AL: "Myristoylated alanine-rich C-kinase substrate protein: A major intracellular regulatory molecule controlling secretion of mucin by human airway goblet cells." CHEST, vol. 117, no. 5 Suppl. 1, May 2000 (2000-05), pages 266S-267S, XP000946039 ISSN: 0012-3692
See also references of EP 1154786A2
NAKAMURA MASATSUGU ET AL: "Mucin-like glycoprotein secretion is mediated by cyclic-AMP and protein kinase C signal transduction pathways in rat corneal epithelium." EXPERIMENTAL EYE RESEARCH, vol. 66, no. 5, May 1998 (1998-05), pages 513-519, XP000946070 ISSN: 0014-4835
THELEN M ET AL: "TUMOR NECROSIS FACTOR ALPHA MODIFIES AGONIST-DEPENDENT RESPONSES IN HUMAN NEUTROPHILS BY INDUCING THE SYNTHESIS AND MYRISTOYLATION OF A SPECIFIC PROTEIN KINASE C SUBSTRATE" PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES, vol. 87, no. 15, 1990, pages 5603-5607, XP002147379 1990 ISSN: 0027-8424
ADLER K B ET AL: "Effects of inflammatory mediators and drugs on mucus secretion and mucociliary function." RESEARCH IN IMMUNOLOGY, vol. 149, no. 3, March 1998 (1998-03), pages 245-248, XP000946047 Meeting on New Therapeutic Approaches for Allergic Diseases of the Respiratory Tract;Paris, France; April 1-4, 1998 ISSN: 0923-2494
ADLER KENNETH B ET AL: "Myristoylated alanine-rich C-kinase substrate protein: A major intracellular regulatory molecule controlling secretion of mucin by human airway goblet cells." CHEST, vol. 117, no. 5 Suppl. 1, May 2000 (2000-05), pages 266S-267S, XP000946039 ISSN: 0012-3692
See also references of EP 1154786A2
Attorney, Agent or Firm:
Biswas, Sorojini J. (Bigel Sibley & Sajove, P.A. P.O. Box 37428 Raleigh NC, US)
Download PDF:
Claims:
We claim:
| 1. | A method of inhibiting mucus secretion by a mucussecreting cell, comprising administering to said cell a mucusinhibitory amount of a compound that inhibits MARCKS proteinrelated mucus secretion, whereby mucus secretion by said cell is reduced compared to that which would occur in the absence of said compound. |
| 2. | A method according to claim 1 wherein said compound is an active fragment of a MARCKS protein. |
| 3. | A method according to claim 2 wherein said fragment has a sequence comprising from about 10 to about 50 contiguous amino acids from SEQ ID NO: 3. |
| 4. | A method according to claim 1 wherein said compound is a peptide having a sequence comprising from about 10 to about 50 contiguous amino acids from SEQ ID NO: 3. |
| 5. | A method according to claim 1 wherein said compound is an antisense oligonucleotide directed against endogenous nucleotide molecules that encode a MARCKS protein. |
| 6. | A method according to claim 1 wherein said compound is selected from the group consisting of okadaic acid, calphostin C, Rp8BrcGMP and LY83583. |
| 7. | A method according to claim 1 wherein said mucussecreting cell is an epithelial cell contained within airway mucous membranes or gastrointestinal mucous membranes. |
| 8. | A method according to claim 1, wherein said compound is administered to the airways of a mammalian subject. |
| 9. | A method according to claim 1, wherein said compound is administered to the gastrointestinal tract of a mammalian subject. |
| 10. | A method according to claim 1 wherein said compound is administered by inhalation. |
| 11. | A method of inhibiting mucus secretion by a mucussecreting cell, comprising administering to said cell a peptide inhibitor of MARCKSrelated mucus secretion, such that mucus secretion by said cell is inhibited compared to that which would occur in the absence of said peptide. |
| 12. | A method according to claim 11 wherein the sequence of said peptide comprises SEQ ID NO: 1 or SEQ ID NO: 2, and said peptide is delivered in a mucusinhibiting amount. |
| 13. | A method according to claim 11 wherein said peptide inhibitor has a sequence comprising from about 10 to about 50 contiguous amino acids from SEQ ID NO: 3. |
| 14. | A method according to claim 11 wherein said mucussecreting cell is an epithelial cell contained within airway mucous membranes or gastrointestinal mucous membranes. |
| 15. | A method according to claim 11, wherein said peptide is administered to the airways of a mammalian subject. |
| 16. | A method according to claim 11, wherein said peptide is administered to the gastrointestinal tract of a mammalian subject. |
| 17. | A method according to claim 15 wherein said peptide is administered by inhalation. |
| 18. | A method of inhibiting mucus secretion in the airways of a subject in need of such treatment, comprising administering to the airways of said subject a mucusinhibiting amount of a compound that inhibits the MARCKSrelated release of mucin, whereby mucus secretion in the airways of the subject is reduced compared to that which would occur in the absence of said treatment. |
| 19. | A method according to claim 18 wherein said subject is a mammalian subject suffering from a condition selected from the group consisting of bronchitis, asthma, cystic fibrosis, chronic obstructive pulmonary disease, emphysema, pneumonia, influenza, rhinitis and the common cold. |
| 20. | A method according to claim 18 wherein said compound is an active fragment of a MARCKS protein. |
| 21. | A method according to claim 20 wherein said active fragment of MARCKS protein comprises a sequence selected from SEQ ID NO: I and SEQ ID NO: 2, and said peptide is delivered in a mucusinhibiting amount. |
| 22. | A method according to claim 20 wherein said peptide has a sequence comprising from about 10 to about 50 contiguous amino acids from SEQ ID NO: 3. |
| 23. | A method according to claim 18 wherein said compound is an antisense oligonucleotide. |
| 24. | A method according to claim 18 wherein said compound is selected from the group consisting of okadaic acid, calphostin C, Rp8BrcGMP) and LY83583. |
| 25. | A method according to claim 18 wherein said compound is administered by inhalation. |
| 26. | A method of increasing mucus secretion by a mucussecreting cell, comprising administering to said cell a secretionenhancing fragment of a MARCKS protein in an amount sufficient to increase mucus secretion by said cell, compared to that which would occur in the absence of said protein fragment. |
| 27. | A method according to claim 26 wherein said mucussecreting cell is an epithelial cell contained within airway mucous membranes, gastrointestinal mucous membranes, genitourinary membranes, or ocular mucous membranes. |
| 28. | A method according to claim 26 wherein said peptide has a sequence comprising SEQ ID NO: 2, and said peptide is delivered in a mucus enhancing amount. |
| 29. | A method according to claim 26 wherein said peptide has a sequence comprising from about 10 to about 50 contiguous amino acids from SEQ ID NO: 3. |
| 30. | A method according to claim 26, wherein said compound is administered to the airways of a mammalian subject. |
| 31. | A method according to claim 26, wherein said compound is administered to the gastrointestinal tract of a mammalian subject. |
| 32. | A method according to claim 30 wherein said compound is administered by inhalation. |
| 33. | A method according to claim 26 wherein said compound is administered to the eye of a mammalian subject. |
| 34. | A method according to claim 26 wherein said compound is topically administered to the vaginal epithelium. |
| 35. | A method of inhibiting mucus secretion by a mucussecreting cell, comprising administering to said cell a mucusinhibiting amount of an antisense construct that specifically binds to endogenous MARCKS protein encoding sequences under physiological conditions, wherein mucus secretion by said cell is inhibited compared to that which would occur in the absence of such administration. |
| 36. | A method according to claim 35 wherein said antisense construct is introduced into said cells via aerosol administration to the lungs. |
| 37. | A method according to claim 35 wherein said antisense construct is introduced into said cells in a liposome. |
| 38. | A method according to claim 35 wherein said mucussecreting cell is an epithelial cell contained within airway mucous membranes or gastrointestinal mucous membranes. |
| 39. | A pharmaceutical formulation comprising a mucusinhibiting peptide fragment of MARCKS, and a pharmaceutically acceptable carrier. |
| 40. | A pharmaceutical formulation according to claim 39, wherein said peptide has an amino acid sequence comprising SEQ ID NO: 1 or SEQ ID NO: 2. |
| 41. | A pharmaceutical formulation according to claim 39 wherein said peptide has a sequence comprising from about 10 to about 50 contiguous amino acids from SEQ ID NO: 3. |
| 42. | A pharmaceutical formulation according to claim 39 where said composition is aerosolized. |
| 43. | A pharmaceutical formulation according to claim 39 where said peptides are contained within liposomes. |
| 44. | An oligonucleotide consisting of about 10 to 50 nucleotides having a nucleotide sequence that hybridizes to nucleotide molecules encoding a MARCKS protein under physiologic conditions, and wherein said oligonucleotide inhibits expression of said MARCKS protein when administered to a cell containing said endogenous nucleotide molecules. |
| 45. | A pharmaceutical formulation comprising an oligonucleotide according to claim 44 and a pharmaceutically acceptable carrier. |
| 46. | A pharmaceutical formulation of claim 45 where said composition is aerosolized. |
| 47. | A pharmaceutical formulation of claim 45 wherein said oligonucleotide is carried within a liposome. |
| 48. | A method of inhibiting mucus secretion by a mucussecreting cell, comprising administering to said cell a mucusinhibitory amount of a compound that binds to a target site selected from: (a) mucin granule membranes at the site bound by MARCKS protein; and 5 (b) MARCKS protein at the mucin granule binding site; wherein the amount of mucus secreted by said cell is reduced compared to that which would occur in the absence of said compound. |
| 49. | A method according to claim 48 where said compound is a peptide having an amino acid sequence that comprises from about 10 to about 50 contiguous amino acids from an Nterminal sequence of a MARCKS protein. |
| 50. | A method according to claim 49 where said peptide is myristoylated. |
| 51. | A method of enhancing mucus secretion by a mucussecreting cell, comprising administering to said cell a mucusenhancing amount of a compound that binds to an endogenous inhibitor of MARCKS protein, and wherein the amount of mucus secreted by said cell is increased compared to that which would 5 occur in the absence of said compound. |
| 52. | A method according to claim 51 where said compound is a peptide having an amino acid sequence that comprises from about 10 to about 50 contiguous amino acids from a phosphorylation site domain of a MARCKS protein. |
| 53. | A method according to claim 52 where said peptide is myristoylated. |
| 54. | The method according to Claim 52 wherein the compound is calmodulin. |
| 55. | A method of screening a test compound for the ability to bind, in a mucussecreting cell, to a site selected from (a) mucin granule membranes at the site bound by MARCKS protein, and (b) a MARCKS protein at the mucin granule membrane binding site, said method comprising administering said test compound to a mucussecreting cell and then detecting whether said test compound inhibits binding of endogenous MARCKS protein to the mucin granule membrane. |
| 56. | A method according to claim 55 where said cells are normal human bronchial epithelial cells. |
| 57. | A method according to claim 55, further comprising administering a compound known to stimulate mucus secretion by said cell. |
| 58. | A method according to claim 57 where said compound known to stimulate mucus secretion is selected from the group consisting of uridine 5' triphosphate (UTP), PMA, and 8bromocGMP. |
| 59. | A method according to claim 55 where said test compound is selected from the group consisting of a peptide fragment of a MARCKS protein, and peptide analogs thereof. |
| 60. | A method according to claim 55 where said test compound is labelled with a detectable molecule. |
| 61. | A method according to claim 55 where said test compound is an antibody that specifically binds to MARCKS protein. |
| 62. | A method of enhancing mucus secretion by a mucus secreting cell, comprising administering to the cell a mucus enhancing amount of a compound that increases the amount of MARCKS protein in the cell, wherein mucus secretion is enhanced in comparison to that which would occur in the absence of the compound. |
| 63. | A method according to claim 62, wherein the compound is a peptide. |
| 64. | A method according to claim 62, wherein the compound is a cytokine. |
| 65. | A method according to claim 62, wherein the compound is TNFa. |
| 66. | A method according to claim 62, wherein the compound is lipopolysaccharide (LPS). |
Description:
INTERNATIONALSEARCH REPORT Int ional AppilodlonNo PCT/US00/05050 C, (Continuation) DOCUMENTS CONSIDEREDTOBERELEVANT Category° Citationofdocument,withindication,whereappropnate,oftherelev
antpassagesRelevanttodaimNo. ADRAY-CHARIERNATHALIEETAL:"Regulation 1-43, ofmucinsecretioninhumangallbladder48-61 epithelialcells:Predominantroleof calciumandproteinkinaseC." GASTROENTEROLOGY, vol.112,no.3,1997,pages978-990, XP000946031 ISSN:0016-5085 thewholedocument ANAKAMURA MASATSUGU ET AL :"Mucin-like1-43, glycoproteinsecretionismediatedby48-61 cyclic-AMPandproteinkinaseCsignal transductionpathwaysinratcorneal epithelium." EXPERIMENTALEYERESEARCH, vol.66,no.5,May1998(1998-05),pages 513-519,XP000946070 ISSN:0014-4835 thewholedocument ATHELENMETAL:"TUMORNECROSISFACTOR62-66 ALPHAMODIFIESAGONIST-DEPENDENTRESPONSES INHUMANNEUTROPHILSBYINDUCINGTHE SYNTHESISANDMYRISTOYLATIONOFASPECIFIC PROTEINKINASECSUBSTRATE" PROCEEDINGSOFTHENATIONALACADEMYOF SCIENCESOFTHEUNITEDSTATES, vol.87,no.15,1990,pages5603-5607, XP002147379 1990 ISSN:0027-8424 thewholedocument AADLERKBETAL:"Effectsofinflammatory6,24, mediatorsanddrugsonmucussecretionand62-66 mucociliaryfunction." RESEARCHINIMMUNOLOGY, vol.149,no.3,March1998(1998-03), pages245-248,XP000946047 MeetingonNewTherapeuticApproachesfor AllergicDiseasesoftheRespiratory Tract;Paris,France;April1-4,1998 ISSN:0923-2494 thewholedocument 2 2 INTERNATIONALSEARCHREPORT irit Jonal ApplicationNo PCT/US00/05050 C.(Continuation) DOCUMENTS CONSIDEREDTOBERELEVANT Category° Citation ofdocument,withindication,whereappropriate,oftherelevantpass
agesRelevanttodaimNo. TADLERKENNETHBETAL:"Myristoylated 1-61 alanine-richC-kinasesubstrateprotein:A majorintracellularregulatorymolecule controllingsecretionofmucinbyhuman airwaygobletcells." CHEST, vol.117,no.5Suppl.1, May2000(2000-05),pages266S-267S, XP000946039 ISSN:0012-3692 thewholedocument 2 INTERNATIONALSEARCHREPORT Irtional ApplicationNo infonnationonpateM tamily members PCT/US00/05050 PatentdocumentPublication Patent family Publication cited in search report date member (s) date WO9300353A07-01-1993AU 2240492 A 25-01-1993 EP 0593580 A 27-04-1994 AU 3665893 A 03-09-1993 WO 9316178 A 19-08-1993WO 9316178 A 19-08-1993 US5858784A12-01-1999AU 1912897 A 26-06-1997 AU 679170 B 26-06-1997 AU 3329193 A 28-07-1993 AU 3467193 A 19-07-1993 CA 2126101 A 24-06-1993 CA 2126103 A 08-07-1993 EP 0619742 A 19-10-1994 EP 0625207 A 23-11-1994 JP 7502510 T 16-03-1995 JP 7505134 T 08-06-1995 US 5641662 A 24-06-1997 WO 9312240 A 24-06-1993 WO 9312756 A 08-07-1993 US 6001644 A 14-12-1999 US 5756353 A 26-05-1998