Methods, Compounds, and Compositions For Treatment and Prophylaxis in the Respiratory Tract

Abstract

The present invention provides a method of reducing the quantity of mucus in the respiratory tract of a subject with elevated levels of mucus in said respiratory tract. The method includes administering to the subject a compound or composition containing a therapeutically effective amount of a fusion protein comprising a sialidase or an active portion thereof and an anchoring domain. The therapeutically effective amount comprises an amount of the fusion protein that results in a reduction of the quantity of mucus in the respiratory tract after administration of the compound or composition when compared to the quantity of mucus present prior to administration of the compound or composition.

What is claimed is: 
     
         1 . A method of reducing the quantity or level of mucus or preventing an increase in the quantity or level of mucus in a respiratory tract of a subject, the method comprising:
 administering to the subject a compound or composition comprising a therapeutically effective amount of a fusion protein, wherein   the fusion protein comprises at least one catalytic domain of a sialidase, wherein the catalytic domain of the sialidase comprises the sequence of amino acids extending from amino acid 274 to amino acid 666 of SEQ ID NO:12, inclusive, and at least one anchoring domain, wherein the anchoring domain is a glycosaminoglycan (GAG) binding domain of human amphiregulin comprising the amino acid sequence of SEQ ID NO:7; and   the therapeutically effective amount comprises an amount of the fusion protein that results in a reduction of the quantity of mucus in the respiratory tract after administration of the compound or composition when compared to the quantity of mucus present prior to administration of the compound or composition.   
     
     
         2 . The method of  claim 1 , wherein the composition further comprises one or more additional compounds. 
     
     
         3 . The method of  claim 2 , wherein the one or more additional compounds comprises MgSO 4  or an analog thereof. 
     
     
         4 . The method of  claim 2 , wherein the one or more additional compounds comprises CaCl 2  or an analog thereof. 
     
     
         5 . The method of  claim 2 , wherein the one or more additional compounds comprises Histidine or an analog thereof. 
     
     
         6 . The method of  claim 2 , wherein the one or more additional compounds comprises Histidine-HCl or an analog thereof. 
     
     
         7 . The method of  claim 2 , wherein the one or more additional compounds comprises Trehalose or an analog thereof. 
     
     
         8 . The method of  claim 2 , wherein the one or more additional compounds comprises MgSO 4 , CaCl 2 , Histidine, Histine-HCl and Trehalose. 
     
     
         9 . The method of  claims 1 - 8 , wherein the compound or composition is formulated as a spray. 
     
     
         10 . The method of  claims 1 - 8 , wherein the compound or composition is formulated as an inhalant. 
     
     
         11 . The method of  claims 1 - 8 , wherein the compound or composition is formulated as a solution for injection. 
     
     
         12 . The method of  claims 1 - 8 , wherein the compound or composition is formulated as a cream, salve, gel, or ointment. 
     
     
         13 . The method of  claims 1 - 8 , wherein the compound or composition is formulated as a pill, tablet, lozenge, suspension, or solution that can be administered orally. 
     
     
         14 . The method of  claims 1 - 8 , wherein said administration is by use of a nasal spray. 
     
     
         15 . The method of  claims 1 - 8 , wherein said administration is by use of an inhaler. 
     
     
         16 . The method of  claims 1 - 8 , wherein said administration is performed from once to four times a day. 
     
     
         17 . The method of  claims 1 - 8 , wherein said administration is by topical application. 
     
     
         18 . The method of  claims 1 - 8 , wherein said administration is by oral administration. 
     
     
         19 . The method of  claims 1 - 8 , wherein said administration is by local or intravenous injection. 
     
     
         20 . The method of  claims 1 - 19 , wherein the subject is human. 
     
     
         21 . The method of  claims 1 - 19 , wherein the subject is a non-human animal. 
     
     
         22 . The method of  claims 1 - 20 , wherein the subject has one or more of the following conditions: chronic obstructive pulmonary disease (COPD), bronchitis, bronchiectasis, cystic fibrosis (CF), vasculitis, mucus plugging, Wegener's granulomatosis, pneumonia, tuberculosis, cancer involving the lungs or the respiratory tract, Kartagener syndrome, Young's syndrome, chronic sinopulmonary infection, alpha 1-antitrypsin deficiency, primary immunodeficiency, acquired immune deficiency syndrome, opportunistic infection, an infectious state, a post infectious state, common cold, exercise induced hypersecretion of mucus, inflammatory bowel disease, ulcerative colitis, Crohn's disease, respiratory infection, respiratory obstruction, inhalation or aspiration of a toxic gas, pulmonary aspiration, or alcoholism. 
     
     
         23 . The method of  claims 1 - 22 , wherein the fusion protein has the sequence set forth in SEQ ID NO:21. 
     
     
         24 . A method of reducing the quantity of mucus in the respiratory tract of a subject with elevated levels of mucus in said respiratory tract, comprising:
 administering to the subject a compound or composition comprising a therapeutically effective amount of a peptide comprising a sialidase or an active portion thereof and, optionally, an anchoring domain, wherein   the therapeutically effective amount comprises an amount of the peptide that results in a reduction of the quantity of mucus in the respiratory tract after administration of the compound or composition when compared to the quantity of mucus present prior to administration of the compound or composition.   
     
     
         25 . The method of  claim 24 , wherein said sialidase or active portion thereof is substantially homologous to  Clostridium perfringens  sialidase or its catalytic domain, substantially homologous to  Actinomyces viscosus  sialidase or its catalytic domain, substantially, homologous to  Arthrobacter ureafaciens  sialidase or its catalytic domain, substantially homologous to  Micromonospora viridifaciens  sialidase or its catalytic domain, substantially homologous to human Neu2 sialidase or its catalytic domain, or substantially homologous to human Neu4 sialidase or its catalytic domain. 
     
     
         26 . The method of  claim 25 , wherein said sialidase or active portion thereof is substantially homologous to  Actinomyces viscosus  sialidase or its catalytic domain. 
     
     
         27 . The method of  claim 24 , wherein the peptide comprises an anchoring domain, wherein said anchoring domain is a glycosaminoglycan (GAG) binding domain. 
     
     
         28 . The method of  claim 27 , wherein said GAG-binding domain is substantially homologous to the GAG-binding domain of human platelet factor 4, substantially homologous to the GAG-binding domain of human interleukin 8, substantially homologous to the GAG-binding domain of human antithrombin III, substantially homologous to the GAG-binding domain of human apoprotein E, substantially homologous to the GAG-binding domain of human angio-associated migratory protein, or substantially homologous to the GAG-binding domain of human amphiregulin. 
     
     
         29 . The method of  claim 28 , wherein said GAG-binding domain is substantially homologous to the GAG-binding domain of human amphiregulin. 
     
     
         30 . The method of  claim 24 , wherein the composition further comprises one or more additional compounds. 
     
     
         31 . The method of  claim 30 , wherein the one or more additional compounds comprises MgSO 4  or an analog thereof. 
     
     
         32 . The method of  claim 30 , wherein the one or more additional compounds comprises CaCl 2  or an analog thereof. 
     
     
         33 . The method of  claim 30 , wherein the one or more additional compounds comprises Histidine or an analog thereof. 
     
     
         34 . The method of  claim 30 , wherein the one or more additional compounds comprises Histidine-HCl or an analog thereof. 
     
     
         35 . The method of  claim 30 , wherein the one or more additional compounds comprises Trehalose or an analog thereof. 
     
     
         36 . The method of  claim 30 , wherein the one or more additional compounds comprises MgSO 4 , CaCl 2 , Histidine, Histine-HCl and Trehalose. 
     
     
         37 . The method of  claims 24 - 36 , wherein the compound or composition is formulated as a spray. 
     
     
         38 . The method of  claims 24 - 36 , wherein the compound or composition is formulated as an inhalant. 
     
     
         39 . The method of  claims 24 - 36 , wherein the compound or composition is formulated as a solution for injection. 
     
     
         40 . The method of  claims 24 - 36 , wherein the compound or composition is formulated as a cream, salve, gel, or ointment. 
     
     
         41 . The method of  claims 24 - 36 , wherein the compound or composition is formulated as a pill, tablet, lozenge, suspension, or solution that can be administered orally. 
     
     
         42 . The method of  claims 24 - 36 , wherein said administration is by use of a nasal spray. 
     
     
         43 . The method of  claim 24 - 36 , wherein said administration is by use of an inhaler. 
     
     
         44 . The method of  claim 24 - 36 , wherein said administration is performed from once to four times a day. 
     
     
         45 . The method of  claim 24 - 36 , wherein said administration is by topical application. 
     
     
         46 . The method of  claim 24 - 36 , wherein said administration is by oral administration. 
     
     
         47 . The method of  claim 24 - 36 , wherein said administration is by local or intravenous injection. 
     
     
         48 . The method of  claim 24 - 47 , wherein the subject is a human. 
     
     
         49 . The method of  claim 24 - 47 , wherein the subject is a non-human animal. 
     
     
         50 . The method of  claims 24 - 48 , wherein the subject has one or more of the following conditions: chronic obstructive pulmonary disease (COPD), bronchitis, bronchiectasis, cystic fibrosis (CF), vasculitis, mucus plugging, Wegener's granulomatosis, pneumonia, tuberculosis, cancer involving the lungs or the respiratory tract, Kartagener syndrome, Young's syndrome, chronic sinopulmonary infection, alpha 1-antitrypsin deficiency, primary immunodeficiency, acquired immune deficiency syndrome, opportunistic infection, an infectious state, a post infectious state, common cold, exercise induced hypersecretion of mucus, inflammatory bowel disease, ulcerative colitis, Crohn's disease, respiratory infection, respiratory obstruction, inhalation or aspiration of a toxic gas, pulmonary aspiration, or alcoholism. 
     
     
         51 . The method of  claims 24 - 50 , wherein the fusion protein has the sequence set forth in SEQ ID NO:21. 
     
     
         52 . A method of limiting an increase in the quantity of mucus in the respiratory tract of a subject above a baseline level of mucus in said subject's respiratory tract, comprising:
 administering to the subject a compound or composition comprising a therapeutically effective amount of a fusion protein, wherein   the fusion protein comprises at least one catalytic domain of a sialidase, wherein the catalytic domain of the sialidase comprises the sequence of amino acids extending from amino acid 274 to amino acid 666 of SEQ ID NO:12, inclusive, and at least one anchoring domain, wherein the anchoring domain is a glycosaminoglycan (GAG) binding domain of human amphiregulin comprising the amino acid sequence of SEQ ID NO:7; and   the therapeutically effective amount comprises an amount of the fusion protein that limits an increase in the quantity of mucus in the respiratory tract of said subject above a baseline level after administration of the compound or composition.   
     
     
         53 . The method of  claim 52 , wherein the composition further comprises one or more additional compounds. 
     
     
         54 . The method of  claim 53 , wherein the one or more additional compounds comprises MgSO 4  or an analog thereof. 
     
     
         55 . The method of  claim 53 , wherein the one or more additional compounds comprises CaCl 2  or an analog thereof. 
     
     
         56 . The method of  claim 53 , wherein the one or more additional compounds comprises Histidine or an analog thereof. 
     
     
         57 . The method of  claim 53 , wherein the one or more additional compounds comprises Histidine-HCl or an analog thereof. 
     
     
         58 . The method of  claim 53 , wherein the one or more additional compounds comprises Trehalose or an analog thereof. 
     
     
         59 . The method of  claim 53 , wherein the one or more additional compounds comprises MgSO 4 , CaCl 2 , Histidine, Histine-HCl and Trehalose. 
     
     
         60 . The method of one or more of  claims 52 - 59 , wherein the compound or composition is formulated as a spray. 
     
     
         61 . The method of  claims 52 - 59 , wherein the compound or composition is formulated as an inhalant. 
     
     
         62 . The method of  claims 52 - 59 , wherein the compound or composition is formulated as a solution for injection. 
     
     
         63 . The method of  claim 52 - 59 , wherein the compound or composition is formulated as a cream, salve, gel, or ointment. 
     
     
         64 . The method of  claims 52 - 59 , wherein the compound or composition is formulated as a pill, tablet, lozenge, suspension, or solution that can be administered orally. 
     
     
         65 . The method of  claims 52 - 59 , wherein said administration is by use of a nasal spray. 
     
     
         66 . The method of  claims 52 - 59 , wherein said administration is by use of an inhaler. 
     
     
         67 . The method of  claims 52 - 59 , wherein said administration is performed from once to four times a day. 
     
     
         68 . The method of  claims 52 - 59 , wherein said administration is by topical application. 
     
     
         69 . The method of  claims 52 - 59 , wherein said administration is by oral administration. 
     
     
         70 . The method of  claims 52 - 59 , wherein said administration is by local or intravenous injection. 
     
     
         71 . The method of  claims 52 - 70 , wherein the subject is human. 
     
     
         72 . The method of  claim 52 - 70 , wherein the subject is a non-human animal. 
     
     
         73 . The method of  claim 52 - 72 , wherein the subject has one or more of the following conditions: chronic obstructive pulmonary disease (COPD), bronchitis, bronchiectasis, cystic fibrosis (CF), vasculitis, mucus plugging, Wegener's granulomatosis, pneumonia, tuberculosis, cancer involving the lungs or the respiratory tract, Kartagener syndrome, Young's syndrome, chronic sinopulmonary infection, alpha 1-antitrypsin deficiency, primary immunodeficiency, acquired immune deficiency syndrome, opportunistic infection, an infectious state, a post infectious state, common cold, exercise induced hypersecretion of mucus, inflammatory bowel disease, ulcerative colitis, Crohn's disease, respiratory infection, respiratory obstruction, inhalation or aspiration of a toxic gas, pulmonary aspiration, or alcoholism. 
     
     
         74 . The method of  claim 52 - 73 , wherein the fusion protein has the sequence set forth in SEQ ID NO:21. 
     
     
         75 . A method of limiting an increase in the quantity of mucus in the respiratory tract of a subject above a baseline level of mucus in said subject's respiratory tract, comprising:
 administering to the subject a compound or composition containing a therapeutically effective amount of a peptide comprising a sialidase or an active portion thereof and, optionally, an anchoring domain, wherein   the therapeutically effective amount comprises an amount of the peptide that limits an increase in the quantity of mucus in the respiratory tract of said subject above a baseline level after administration of the compound or composition.   
     
     
         76 . The method of  claim 75 , wherein said sialidase or active portion thereof is substantially homologous to  Clostridium perfringens  sialidase or its catalytic domain, substantially homologous to  Actinomyces viscosus  sialidase or its catalytic domain, substantially, homologous to  Arthrobacter ureafaciens  sialidase or its catalytic domain, substantially homologous to  Micromonospora viridifaciens  sialidase or its catalytic domain, substantially homologous to human Neu2 sialidase or its catalytic domain, or substantially homologous to human Neu4 sialidase or its catalytic domain. 
     
     
         77 . The method of  claim 76 , wherein said sialidase or active portion thereof is substantially homologous to  Actinomyces viscosus  sialidase or its catalytic domain. 
     
     
         78 . The method of  claim 75 , wherein the peptide comprises an anchoring domain, wherein said anchoring domain is a glycosaminoglycan (GAG) binding domain. 
     
     
         79 . The method of  claim 78 , wherein said GAG-binding domain is substantially homologous to the GAG-binding domain of human platelet factor 4, substantially homologous to the GAG-binding domain of human interleukin 8, substantially homologous to the GAG-binding domain of human antithrombin III, substantially homologous to the GAG-binding domain of human apoprotein E, substantially homologous to the GAG-binding domain of human angio-associated migratory protein, or substantially homologous to the GAG-binding domain of human amphiregulin. 
     
     
         80 . The method of  claim 78 , wherein said GAG-binding domain is substantially homologous to the GAG-binding domain of human amphiregulin. 
     
     
         81 . The method of  claim 75 , wherein the composition further comprises one or more additional compounds. 
     
     
         82 . The method of  claim 71 , wherein the one or more additional compounds comprises MgSO 4  or an analog thereof. 
     
     
         83 . The method of  claim 71 , wherein the one or more additional compounds comprises CaCl 2  or an analog thereof. 
     
     
         84 . The method of  claim 71 , wherein the one or more additional compounds comprises Histidine or an analog thereof. 
     
     
         85 . The method of  claim 71 , wherein the one or more additional compounds comprises Histidine-HCl or an analog thereof. 
     
     
         86 . The method of  claim 71 , wherein the one or more additional compounds comprises Trehalose or an analog thereof. 
     
     
         87 . The method of  claim 71 , wherein the one or more additional compounds comprises MgSO 4 , CaCl 2 , Histidine, Histine-HCl and Trehalose. 
     
     
         88 . The method of  claims 75 - 87 , wherein the compound or composition is formulated as a spray. 
     
     
         89 . The method of  claims 75 - 87 , wherein the compound or composition is formulated as an inhalant. 
     
     
         90 . The method of  claims 75 - 87 , wherein the compound or composition is formulated as a solution for injection. 
     
     
         91 . The method of  claims 75 - 87 , wherein the compound or composition is formulated as a cream, salve, gel, or ointment. 
     
     
         92 . The method of  claims 75 - 87 , wherein the compound or composition is formulated as a pill, tablet, lozenge, suspension, or solution that can be administered orally. 
     
     
         93 . The method of  claims 75 - 87 , wherein said administration is by use of a nasal spray. 
     
     
         94 . The method of  claims 75 - 87 , wherein said administration is by use of an inhaler. 
     
     
         95 . The method of  claims 75 - 87 , wherein said administration is performed from once to four times a day. 
     
     
         96 . The method of  claims 75 - 87 , wherein said administration is by topical application. 
     
     
         97 . The method of  claims 75 - 87 , wherein said administration is by oral administration. 
     
     
         98 . The method of  claims 75 - 87 , wherein said administration is by local or intravenous injection. 
     
     
         99 . The method of  claims 75 - 98 , wherein the subject is a human. 
     
     
         100 . The method of  claims 75 - 98 , wherein the subject is a non-human animal. 
     
     
         101 . The method of  claim 75 - 100 , wherein the subject has one or more of the following conditions: chronic obstructive pulmonary disease (COPD), bronchitis, bronchiectasis, cystic fibrosis (CF), vasculitis, mucus plugging, Wegener's granulomatosis, pneumonia, tuberculosis, cancer involving the lungs or the respiratory tract, Kartagener syndrome, Young's syndrome, chronic sinopulmonary infection, alpha 1-antitrypsin deficiency, primary immunodeficiency, acquired immune deficiency syndrome, opportunistic infection, an infectious state, a post infectious state, common cold, exercise induced hypersecretion of mucus, inflammatory bowel disease, ulcerative colitis, Crohn's disease, respiratory infection, respiratory obstruction, inhalation or aspiration of a toxic gas, pulmonary aspiration, or alcoholism. 
     
     
         102 . The method of  claim 75 - 101 , wherein the fusion protein has the sequence set forth in SEQ ID NO:21. 
     
     
         103 . The method of any one of  claims 1 ,  24 ,  52 , and  75 , further comprising selecting a subject for treatment. 
     
     
         104 . The method of  claim 103 , wherein the subject is not infected with one or more of influenza virus, parainfluenza virus, and/or respiratory syncytial virus (RSV). 
     
     
         105 . The method of  claim 103 , wherein the subject is not infected with one or more of active influenza virus, active parainfluenza virus, and/or active RSV. 
     
     
         106 . The method of  claim 103 , wherein the subject is not suffering from asthma. 
     
     
         107 . The method of  claim 103 , wherein the subject does not have asthma or is not asthmatic. 
     
     
         108 . The method of any one of  claims 1 ,  24 ,  52 ,  75 , and  103 , wherein the subject further has influenza or is infected by influenza virus. 
     
     
         109 . The method of  claim 108 , wherein the compound or composition comprises a protein or peptide having the sequence set forth in SEQ ID NO:21. 
     
     
         110 . The method of any one of  claims 52  and  75 , further comprising determining the baseline level of mucus in the subject's respiratory tract prior to administering to the subject the compound or composition. 
     
     
         111 . The method of  claim 110 , further comprising determining a second level of mucus in the subject's respiratory tract following administering to the subject the compound or composition. 
     
     
         112 . The method of  claim 112 , further comprising continuing treatment until the level or increase in level of mucus in the subject's respiratory tract is decreased or limited in the second level of mucus compared to the baseline level of mucus in the subject's respiratory tract. 
     
     
         113 . A method, comprising:
 (a) administering a collection of compounds or compositions comprising sialidase(s) and/or catalytically active portion(s) thereof to an animal model of asthma;   (b) quantitating the level of mucus in the lung of the animal;   (c) comparing the level measured in (b) with the level of mucus in the lung of the animal prior to administering the collection of compounds or compositions; and   (d) identifying or selecting a compound or composition that reduces the level of mucus relative to the level of mucus in the absence of the compound or composition.   
     
     
         114 . A method, comprising:
 (a) administering a compound or composition comprising a sialidase and/or a catalytically active portion thereof to an animal model of asthma;   (b) quantitating the level of mucus in the lung of the animal;   (c) comparing the level measured in (b) with the level of mucus in the lung of the animal prior to administering the collection of compounds or compositions; and   (d) identifying whether the compound or composition reduces the level of mucus relative to the level of mucus in the absence of the compound or composition.   
     
     
         115 . A method of assessing whether a compound or composition comprising a sialidase and/or catalytically active portion thereof reduces the quantity of mucus in the respiratory tract of a subject, comprising:
 (a) contacting the muscarinic receptors of an animal subject with a compound or composition comprising a sialidase and/or a catalytically active portion thereof;   (b) administering a muscarinic receptor agonist to the subject;   (c) quantitating the airway resistance in the subject;   (d) comparing the airway resistance level measured in (c) with the airway resistance in the absence of contact with the compound or composition;   (e) identifying whether the compound or composition reduces the airway resistance relative to the airway resistance in the absence of contact with the compound or composition; and   (f) if the compound or composition reduces the airway resistance as determined in (e), assessing the compound or composition as one that reduces the quantity of mucus in the respiratory tract of the subject.