Cell membrane translocation of regulated snare inhibitors, compositions therefor, and methods for treatment of disease

Abstract

Compositions and methods of modulating cellular function and treatment of disease in mammals comprising locally administering a regulated SNARE inhibitor and a translocating agent to the mammal. Regulated SNARE inhibitors include clostridial neurotoxins, tetanus neurotoxin and their free light chain portions and IgA protease. Translocating agents include acids, encapsulating vectors, and transduction domains.

1 . A method of modulating cellular function in a mammal comprising locally administering a modulatorily effective amount of a regulated SNARE inhibitor and a translocating agent to the mammal, whereby translocation of the regulated SNARE inhibitor is facilitated.  
   
   
       2 . The method of  claim 1 , wherein the translocating agent is an acid, a low ionic strength solution, an encapsulating vector, or a transduction domain.  
   
   
       3 . The method of  claim 1 , wherein the regulated SNARE inhibitor is administered in a pharmaceutically acceptable formulation.  
   
   
       4 . The method of  claim 1 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       5 . The method of  claim 4 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       6 . The method of  claim 5 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       7 . The method of  claim 6 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       8 . The method of  claim 4 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       9 . The method of  claim 1 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       10 . The method according to  claim 1  of modulating cellular function in a mammal comprising locally administering an modulatorily effective amount of a regulated SNARE inhibitor to the mammal at a selected site and decreasing a pH value at the selected site.  
   
   
       11 . The method of  claim 10 , wherein the regulated SNARE inhibitor is administered in a pharmaceutically acceptable formulation.  
   
   
       12 . The method of  claim 10 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       13 . The method of  claim 12 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       14 . The method of  claim 13 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       15 . The method of  claim 14 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       16 . The method of  claim 12 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       17 . The method of  claim 10 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       18 . The method of  claim 10 , wherein the pH value is decreased from about 7.4 to within a range of about 3.5 to about 7.  
   
   
       19 . The method of  claim 10 , wherein the pH value is decreased from about 7.4 to within a range of about 3.5 to about 4.5.  
   
   
       20 . The method of  claim 10 , wherein the pH value is decreased from about 7.4 to within a range of about 4.5 to about 6.  
   
   
       21 . The method of  claim 10 , wherein decreasing the pH value comprises locally administering a pharmaceutically acceptable acid to the selected site.  
   
   
       22 . The method of  claim 10 , wherein decreasing the pH value is performed a period of time after administering the regulated SNARE inhibitor.  
   
   
       23 . The method of  claim 10 , wherein the regulated SNARE inhibitor is administered together with or in a pharmaceutically acceptable acidic formulation.  
   
   
       24 . The method of  claim 23 , wherein the pharmaceutically acceptable acidic formulation has a pH value of about 3.5 to about 7.  
   
   
       25 . The method of  claim 23 , wherein the pharmaceutically acceptable acidic formulation has a pH value of about 3.5 to about 4.5.  
   
   
       26 . The method of  claim 23 , wherein the pharmaceutically acceptable acidic formulation has a pH value of about 4.5 to about 6.  
   
   
       27 . The method of  claim 10 , wherein decreasing the pH value comprises applying electrical stimulation to the mammal.  
   
   
       28 . The method of  claim 27 , wherein applying electrical stimulation comprises applying an anode of a DC anode/cathode pair proximal to the selected site.  
   
   
       29 . The method of  claim 27 , wherein the electrical stimulation is applied to an area of the selected site.  
   
   
       30 . The method according to  claim 1  of modulating cellular function in a mammal coinprising locally administering an modulatorily effective amount of a regulated SNARE inhibitor to the mammal at a selected site, by way of a protein transduction domain.  
   
   
       31 . The method of  claim 30 , wherein the regulated SNARE inhibitor is administered in a pharmaceutically acceptable formulation.  
   
   
       32 . The method of  claim 30 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       33 . The method of  claim 32 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       34 . The method of  claim 33 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       35 . The method of  claim 34 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       36 . The method of  claim 32 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       37 . The method of  claim 30 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       38 . The method according to  claim 1  of modulating cellular function in a mammal comprising locally administering an modulatorily effective amount of a regulated SNARE inhibitor to the mammal at a selected site, by way of an encapsulation vector.  
   
   
       39 . The method of  claim 38 , wherein the regulated SNARE inhibitor is administered in a pharmaceutically acceptable formulation.  
   
   
       40 . The method of  claim 38 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       41 . The method of  claim 40 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       42 . The method of  claim 41 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       43 . The method of  claim 42 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       44 . The method of  claim 40 , wherein the regulated SNARE inhibitor is the light chain portion of the bacterial neurotoxin.  
   
   
       45 . The method of  claim 38 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       46 . The method of  claim 38 , wherein the encapsulation vector is a vesicle.  
   
   
       47 . The method of  claim 38 , wherein the encapsulation vector is a liposome.  
   
   
       48 . The method of  claim 38 , wherein the encapsulation vector is a niosome.  
   
   
       49 . The method of  claim 38 , wherein the encapsulation vector is a transferosome.  
   
   
       50 . The method of  claim 38 , wherein the encapsulation vector is a nanoparticle.  
   
   
       51 . The method of  claim 1  of modulating cellular function of a mammal suffering from a disease or medical condition, whereby the disease or medical condition is treated.  
   
   
       52 . The method of  claim 51  of modulating cellular function of a mammal suffering from a disease or medical condition comprising locally administering a therapeutically effective amount of a regulated SNARE inhibitor to a mammal in need of such treatment at a selected site and decreasing a pH value at the selected site.  
   
   
       53 . The method of  claim 52 , wherein the disease or medical condition is pain; inflammation; a migraine headache; allergy; cystic fibrosis; a disease related to adipose tissue; a viral infection; cancer; fever; a disease related to sweating, eccrine, and apocrine; a disease associated with holocrine secretions; acne; a disease relate to mucous secretion; prostatic hypertrophy; a diseases treatable by gene therapy; a disease of the veins; venous stasis, varicose veins; hemorrhoids; or high blood pressure.  
   
   
       54 . The method of  claim 52 , wherein the regulated SNARE inhibitor is administered in a pharmaceutically acceptable formulation.  
   
   
       55 . The method of  claim 52 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       56 . The method of  claim 55 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       57 . The method of  claim 56 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       58 . The method of  claim 57 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       59 . The method of  claim 55 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       60 . The method of  claim 52 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       61 . The method of  claim 52 , wherein the pH value is decreased from about 7.4 to within a range of about 3.5 to about 7.  
   
   
       62 . The method of  claim 52 , wherein the pH value is decreased from about 7.4 to within a range of about 3.5 to about 4.5.  
   
   
       63 . The method of  claim 52 , wherein the pH value is decreased from about 7.4 to within a range of about 4.5 to about 6.  
   
   
       64 . The method of  claim 52 , wherein decreasing the pH value comprises locally administering a pharmaceutically acceptable acid to the selected site.  
   
   
       65 . The method of  claim 52 , wherein decreasing the pH value is performed a period of time after administering the regulated SNARE inhibitor.  
   
   
       66 . The method of  claim 52 , wherein the regulated SNARE inhibitor is administered together with or in a pharmaceutically acceptable acidic formulation.  
   
   
       67 . The method of  claim 66 , wherein the pharmaceutically acceptable acidic formulation has a pH value of about 3.5 to about 7.  
   
   
       68 . The method of  claim 66 , wherein the pharmaceutically acceptable acidic formulation has a pH value of about 3.5 to about 4.5.  
   
   
       69 . The method of  claim 66 , wherein the pharmaceutically acceptable acidic formulation has a pH value of about 4.5 to about 6.  
   
   
       70 . The method of  claim 52 , wherein decreasing the pH value comprises applying electrical stimulation to the mammal.  
   
   
       71 . The method of  claim 70 , wherein applying electrical stimulation comprises applying an anode of a DC anode/cathode pair proximal to the selected site.  
   
   
       72 . The method of  claim 70 , wherein the electrical stimulation is applied to an area of the selected site.  
   
   
       73 . The method of  claim 51  of modulating cellular function of a mammal suffering from a disease, malfunction, or dysfunction comprising locally administering a therapeutically effective amount of a regulated SNARE inhibitor to a mammal in need of such treatment at a selected site by way of a protein transduction domain.  
   
   
       74 . The method of  claim 73 , wherein the disease or medical condition is pain; inflammation; a migraine headache; allergy; cystic fibrosis; a disease related to adipose tissue; a viral infection; cancer; fever; a disease related to sweating, eccrine, and apocrine; a disease associated with holocrine secretions; acne; a disease relate to mucous secretion; prostatic hypertrophy; a diseases treatable by gene therapy; a disease of the veins; venous stasis, varicose veins; hemorrhoids; or high blood pressure.  
   
   
       75 . The method of  claim 73 , wherein the regulated SNARE inhibitor is administered in a pharmaceutically acceptable formulation.  
   
   
       76 . The method of  claim 73 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       77 . The method of  claim 76 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       78 . The method of  claim 77 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       79 . The method of  claim 78 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       80 . The method of  claim 76 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       81 . The method of  claim 73 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       82 . The method of  claim 51  of modulating cellular function of a mammal suffering from a disease, malfunction, or dysfunction comprising locally administering a therapeutically effective amount of a regulated SNARE inhibitor to a mammal in need of such treatment at a selected site by way of an encapsulation vector.  
   
   
       83 . The method of  claim 82 , wherein the disease or medical condition is pain; inflammation; a migraine headache; allergy; cystic fibrosis; a disease related to adipose tissue; a viral infection; cancer; fever; a disease related to sweating, eccrine, and apocrine; a disease associated with holocrine secretions; acne; a disease relate to mucous secretion; prostatic hypertrophy; a diseases treatable by gene therapy; a disease of the veins; venous stasis, varicose veins; hemorrhoids; or high blood pressure.  
   
   
       84 . The method of  claim 82 , wherein the regulated SNARE inhibitor is administered in a pharmaceutically acceptable formulation.  
   
   
       85 . The method of  claim 82 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       86 . The method of  claim 85 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       87 . The method of  claim 86 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       88 . The method of  claim 87 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       89 . The method of  claim 85 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       90 . The method of  claim 82 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       91 . The method of  claim 82 , wherein the encapsulation vector is a vesicle.  
   
   
       92 . The method of  claim 82 , wherein the encapsulation vector is a liposome.  
   
   
       93 . The method of  claim 82 , wherein the encapsulation vector is a niosome.  
   
   
       94 . The method of  claim 82 , wherein the encapsulation vector is a transferosome.  
   
   
       95 . The method of  claim 82 , wherein the encapsulation vector is a nanoparticle.  
   
   
       96 . A pharmaceutical formulation for modulating cellular function in a mammal comprising a therapeutically effective amount of a regulated SNARE inhibitor and a translocating agent in a pharmaceutically acceptable carrier for local delivery.  
   
   
       97 . The pharmaceutical formulation of  claim 96 , wherein the translocating agent is an acid, an acidic environment, an encapsulating vector, or a transduction domain.  
   
   
       98 . The pharmaceutical formulation of  claim 96 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       99 . The pharmaceutical formulation of  claim 98 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       100 . The pharmaceutical formulation of  claim 99 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       101 . The pharmaceutical formulation of  claim 100 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       102 . The pharmaceutical formulation of  claim 96 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       103 . The pharmaceutical formulation of  claim 96 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       104 . The use of a regulated SNARE inhibitor in the preparation of a medicament for a method of treating a disease or medical condition in a mammal comprising locally administering a therapeutically effective amount of a regulated SNARE inhibitor to a mammal in need of such treatment at a selected site related to the disease, by way of acid mediated translocation, a protein transduction domain, or an encapsulation vector.  
   
   
       105 . The use of  claim 104 , wherein the disease or medical condition is pain or inflammation; a migraine headache; allergy; cystic fibrosis; a disease related to adipose tissue; a viral infection; cancer; fever; a disease related to sweating, eccrine, and apocrine; a disease associated with holocrine secretions; acne; a disease relate to mucous secretion; prostatic hypertrophy; a diseases treatable by gene therapy; a disease of the veins; venous stasis, varicose veins; hemorrhoids; or high blood pressure.  
   
   
       106 . The use of  claim 104 , wherein the regulated SNARE inhibitor is a bacterial neurotoxin.  
   
   
       107 . The use of  claim 106 , wherein the bacterial neurotoxin is a clostridial neurotoxin.  
   
   
       108 . The use of  claim 107 , wherein the clostridial neurotoxin is a  botulinum  neurotoxin serotype or tetanus neurotoxin.  
   
   
       109 . The use of  claim 108 , wherein the  botulinum  neurotoxin serotype is A, B, C1, D, E, F or G.  
   
   
       110 . The use of  claim 104 , wherein the regulated SNARE inhibitor is the free light chain portion of the bacterial neurotoxin.  
   
   
       111 . The use of  claim 104 , wherein the regulated SNARE inhibitor is IgA protease.  
   
   
       112 . The use of  claim 104 , wherein the encapsulation vector is a vesicle.  
   
   
       113 . The use of  claim 104 , wherein the encapsulation vector is a liposome.  
   
   
       114 . The use of  claim 104 , wherein the encapsulation vector is a niosome.  
   
   
       115 . The use of  claim 104 , wherein the encapsulation vector is a transferosome.  
   
   
       116 . The use of  claim 104 , wherein the encapsulation vector is a nanoparticle.