Methods and compounds for the treatment of mucus hypersecretion
A method of treating mucus hypersecretion, the causative factor in chronic obstructive pulmonary disease (COPD), asthma and other clinical conditions involving COPD, comprises administering a compound that inhibits exocytosis in mucus secreting cells or neurones that control or direct mucus secretion. Also described is a compound, for use in the treatment of hypersecretion of mucus, which inhibits mucus secretion by inhibiting mucus secretion by mucus secreting cells, and/or inhibiting neurotransmitter release from neuronal cells controlling or directing mucus secretion.
1 - 11 . (canceled)
12 . A single-chain fusion protein comprising:
(a) a light chain (L-chain) or L-chain fragment of a clostridial neurotoxin, which L-chain or L-chain fragment includes the active proteolytic enzyme domain of the L-chain;
(b) a targeting domain comprising or consisting of a growth factor;
(c) a translocating domain that translocates the L-chain or L-chain fragment into the target cell; and
(d) a site for cleavage by a proteolytic enzyme.
13 . A fusion protein according to claim 12 , wherein the growth factor is an epidermal growth factor (EGF).
14 . A fusion protein according to claim 12 , wherein the cleavage site has been introduced into the fusion protein.
15 . A fusion protein according to claim 12 , wherein the cleavage site is selected from the group consisting of: IEGR; DDDDK; EXXYXQSG; LEVLFQGP; LVPRGS; HY; and YH.
16 . A fusion protein according to claim 12 , wherein said cleavage site is not present in a native clostridial neurotoxin.
17 . A fusion protein according to claim 12 , wherein said cleavage site is located between the L-chain or L-chain fragment and the translocating domain.
18 . A fusion protein according to claim 12 , wherein the cleavage site is cleavable by a proteolytic enzyme selected from the group consisting of: factor Xa; enterokinase; TEV protease; precission; thrombin; and genenase.
19 . A nucleic acid encoding a fusion protein according to claim 12 .
20 . A pharmaceutical composition for topical administration to a patient suffering from mucus hypersecretion, comprising:
(i) a polypeptide in an amount effective to inhibit mucus hypersecretion, wherein the polypeptide comprises:
(a) a light chain (L-chain) or L-chain fragment of a clostridial neurotoxin, which L-chain or L-chain fragment includes the active proteolytic enzyme domain of the L-chain;
(b) a targeting domain comprising or consisting of a growth factor; and
(c) a translocating domain that translocates the L-chain or L-chain fragment into the target cell; and
(ii) a formulation component selected from the group consisting of an excipient, an adjuvant and a propellant.
21 . A method of treating hypersecretion of mucus, comprising administering to a patient in need thereof, a therapeutically effective amount of a polypeptide comprising:
(a) a light chain (L-chain) or L-chain fragment of a clostridial neurotoxin, which L-chain or L-chain fragment includes the active proteolytic enzyme domain of the L-chain;
(b) a targeting domain comprising or consisting of a growth factor; and
(c) a translocating domain that translocates the L-chain or L-chain fragment into the target cell;
wherein following said administration, hypersecretion of mucus is reduced.
22 . A method of treating chronic obstructive pulmonary disease (COPD), comprising administering to a patient in need thereof, a therapeutically effective amount of a polypeptide comprising:
(a) a light chain (L-chain) or L-chain fragment of a clostridial neurotoxin, which L-chain or L-chain fragment includes the active proteolytic enzyme domain of the L-chain;
(b) a targeting domain comprising or consisting of a growth factor; and
(c) a translocating domain that translocates the L-chain or L-chain fragment into the target cell;
wherein following said administration, hypersecretion of mucus is reduced.
23 . A method of treating asthma, comprising administering to a patient in need thereof, a therapeutically effective amount of a polypeptide comprising:
(a) a light chain (L-chain) or L-chain fragment of a clostridial neurotoxin, which L-chain or L-chain fragment includes the active proteolytic enzyme domain of the L-chain;
(b) a targeting domain comprising or consisting of a growth factor; and
(c) a translocating domain that translocates the L-chain or L-chain fragment into the target cell;
wherein following said administration, hypersecretion of mucus is reduced.
24 . A method for activating a single-chain fusion protein, comprising:
(i) providing a single-chain fusion protein comprising:
(a) a light chain (L-chain) or L-chain fragment of a clostridial neurotoxin, which L-chain or L-chain fragment includes the active proteolytic enzyme domain of the L-chain;
(b) a targeting domain comprising or consisting of a growth factor;
(c) a translocating domain that translocates the L-chain or L-chain fragment into the target cell; and
(d) a site for cleavage by a proteolytic enzyme;
(ii) contacting said single-chain fusion protein with a proteolytic enzyme that cleaves said site for cleavage; and
(iii) cleaving said single-chain fusion protein at said site for cleavage, and thereby providing a di-chain polypeptide wherein (a) and (c) are linked together by a disulphide bond.
25 . A method according to claim 24 , wherein the proteolytic enzyme is selected from the group consisting of: factor Xa; enterokinase; TEV protease; precission; thrombin; and genenase.
26 . A method according to claim 24 , wherein the cleavage site is selected from the group consisting of: IEGR; DDDK; EXXYXQSG; LEVLFQGP; LVPRGS; HY; and YH.