EP2038299A2 - Modifizierte clostridientoxine mit verbesserten translokationsfähigkeiten und geänderter targeting-aktivität für nichtclostridientoxin-targetzellen - Google Patents
Modifizierte clostridientoxine mit verbesserten translokationsfähigkeiten und geänderter targeting-aktivität für nichtclostridientoxin-targetzellenInfo
- Publication number
- EP2038299A2 EP2038299A2 EP07812774A EP07812774A EP2038299A2 EP 2038299 A2 EP2038299 A2 EP 2038299A2 EP 07812774 A EP07812774 A EP 07812774A EP 07812774 A EP07812774 A EP 07812774A EP 2038299 A2 EP2038299 A2 EP 2038299A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- amino acids
- bont
- seq
- clostridial toxin
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/33—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Clostridium (G)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
- A61P21/02—Muscle relaxants, e.g. for tetanus or cramps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/04—Centrally acting analgesics, e.g. opioids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/50—Fusion polypeptide containing protease site
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/55—Fusion polypeptide containing a fusion with a toxin, e.g. diphteria toxin
Definitions
- FIG. 5 shows modified Clostridial toxins with an enhanced translocation capability and an altered targeting activity located between two other domains.
- FIG. 5A depicts the single polypeptide form of a modified Clostridial toxin with an amino to carboxyl linear organization comprising an enzymatic domain, an altered targeting domain, a translocation domain and a translocation facilitating domain, with the di- chain loop region depicted by the double SS bracket.
- a proteolytic cleavage site (P) within a di-chain loop region is located between the enzymatic and targeting domains. Upon proteolytic cleavage with a P protease, the single chain form of the toxin is converted to the di-chain form.
- P proteolytic cleavage site
- the H N domain also forms a long unstructured loop called the 'translocation belt,' which wraps around a large negatively charged cleft of the light chain that blocks access of the zinc atom to the catalytic-binding pocket of active site.
- the H c domain comprises two distinct structural features of roughly equal size that indicate function. The first, designated the H CN domain, is located in the amino half of the H c domain. The H CN domain forms a ⁇ -barrel, jelly-roll fold. The H C c domain is the second domain that comprises the H c domain.
- a conservative Clostridial toxin light chain variant can function in substantially the same manner as the reference Clostridial toxin light chain on which the conservative Clostridial toxin light chain variant is based, and can be substituted for the reference Clostridial toxin light chain in any aspect of the present invention.
- a BoNT/G light chain comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 non-contiguous amino acid deletions relative to amino acids 1 -446 of SEQ ID NO: 7.
- a BoNT/G light chain comprises a polypeptide having, e.g., at least one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 non-contiguous amino acid deletions relative to amino acids 1 -446 of SEQ ID NO: 7.
- a TeNT light chain comprises a polypeptide having, e.g., at least 70% amino acid identity with amino acids 1 -457 of SEQ ID NO: 8, at least 75% amino acid identity with amino acids 1 -457 of SEQ ID NO: 8, at least 80% amino acid identity with amino acids 1 -457 of SEQ ID NO: 8, at least 85% amino acid identity with amino acids 1 -457 of SEQ ID NO: 8, at least 90% amino acid identity with amino acids 1 -457 of SEQ ID NO: 8 or at least 95% amino acid identity with amino acids 1 -457 of SEQ ID NO: 8.
- the term "conservative Clostridial toxin H N region variant” means a Clostridial toxin H N region that has at least one amino acid substituted by another amino acid or an amino acid analog that has at least one property similar to that of the original amino acid from the reference Clostridial toxin H N region sequence (Table 1 ).
- properties include, without limitation, similar size, topography, charge, hydrophobicity, hydrophilicity, lipophilicity, covalent-bonding capacity, hydrogen-bonding capacity, a physicochemical property, of the like, or any combination thereof.
- a Clostridial toxin translocation domain comprises a non-naturally occurring Clostridial toxin H N region variant, such as, e.g., a conservative Clostridial toxin H N region variant, a non-conservative Clostridial toxin H N region variant, a Clostridial toxin chimeric H N region, an active Clostridial toxin H N region fragment, or any combination thereof.
- a Clostridial toxin translocation domain comprises a BoNT/A H N region.
- a BoNT/A H N region comprises amino acids 449-873 of SEQ ID NO: 1 .
- a BoNT/A H N region comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 contiguous amino acid substitutions relative to amino acids 449-873 of SEQ ID NO: 1 .
- a BoNT/A H N region comprises a polypeptide having, e.g., at least one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 contiguous amino acid substitutions relative to amino acids 449-873 of SEQ ID NO: 1.
- a BoNT/D H N region comprises a polypeptide having, e.g., at least 70% amino acid identity with amino acids 446-864 of SEQ ID NO: 4, at least 75% amino acid identity with amino acids 446-864 of SEQ ID NO: 4, at least 80% amino acid identity with amino acids 446-864 of SEQ ID NO: 4, at least 85% amino acid identity with amino acids 446-864 of SEQ ID NO: 4, at least 90% amino acid identity with amino acids 446-864 of SEQ ID NO: 4 or at least 95% amino acid identity with amino acids 446-864 of SEQ ID NO: 4.
- a TeNT H N region comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 contiguous amino acid additions relative to amino acids 458-881 of SEQ ID NO: 8.
- a TeNT H N region comprises a polypeptide having, e.g., at least one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 contiguous amino acid additions relative to amino acids 458-881 of SEQ ID NO: 8.
- a non-conservative Clostridial toxin H CN region variant can function in substantially the same manner as the reference Clostridial toxin H CN region on which the non-conservative Clostridial toxin H CN region variant is based, and can be substituted for the reference Clostridial toxin H CN region in any aspect of the present invention.
- a non-conservative Clostridial toxin H CN region variant can delete one or more amino acids, two or more amino acids, three or more amino acids, four or more amino acids, five or more amino acids, and ten or more amino acids from the reference Clostridial toxin H CN region on which the non-conservative Clostridial toxin H CN region variant is based.
- a BoNT/D H CN region comprises amino acids 865-1098 of a non-naturally occurring BoNT/D H CN region variant of SEQ ID NO: 4, such as, e.g., amino acids 865-1098 of a conservative BoNT/D H CN region variant of SEQ ID NO: 4, amino acids 865-1098 of a non-conservative BoNT/D H CN region variant of SEQ ID NO: 4, amino acids 865-1098 of an active BoNT/D H CN region fragment of SEQ ID NO: 4, or any combination thereof.
- Non-limiting examples of a conservative enveloped virus fusogenic peptide domain variant include, e.g., conservative influenzavirus fusogenic peptide domain variants, conservative alphavirus fusogenic peptide domain variants, conservative vesiculovirus fusogenic peptide domain variants, conservative respirovirus fusogenic peptide domain variants, conservative morbillivirus fusogenic peptide domain variants, conservative avulavirus fusogenic peptide domain variants, conservative henipavirus fusogenic peptide domain variants, conservative metapneumovirus fusogenic peptide domain variants and conservative foamy virus fusogenic peptide domain variants.
- an influenzavirus fusogenic peptide domain comprises a naturally occurring influenzavirus fusogenic peptide domain variant of SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197 or SEQ ID NO: 198, such as, e.g., an influenzavirus fusogenic peptide domain isoform of SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197 or SEQ ID NO: 198 or an influenzavirus fusogenic peptide domain subtype of SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197 or SEQ ID NO: 198.
- an influenzavirus fusogenic peptide domain comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine or 10 contiguous amino acid substitutions relative to SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197 or SEQ ID NO: 198.
- SEQ ID NO: 232 SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235 or SEQ ID NO: 236, at most 85% amino acid identity with SEQ ID NO: 226, SEQ ID NO: 227, SEQ ID NO: 228, SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231 , SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235 or SEQ ID NO: 236, at most 90% amino acid identity with SEQ ID NO: 226, SEQ ID NO: 227, SEQ ID NO:
- a Clostridial toxin translocation facilitating domain comprises a morbillivirus fusogenic peptide domain.
- a morbillivirus fusogenic peptide domain comprises a naturally occurring morbillivirus fusogenic peptide domain variant, such as, e.g., a morbillivirus fusogenic peptide domain isoform or a morbillivirus fusogenic peptide domain subtype.
- a morbillivirus fusogenic peptide domain comprises a naturally occurring morbillivirus fusogenic peptide domain variant of SEQ ID NO: 244, SEQ ID NO: 245, SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 250, SEQ ID NO: 251 , SEQ ID NO: 252 or SEQ ID NO: 253, such as, e.g., a morbillivirus fusogenic peptide domain isoform of SEQ ID NO: 244, SEQ ID NO: 245, SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 250, SEQ ID NO: 251 , SEQ ID NO: 252 or SEQ ID NO: 253 or a morbillivirus fusogenic peptide domain subtype of SEQ ID NO: 244, SEQ ID NO: 245, SEQ ID NO: 245, SEQ ID NO:
- a morbillivirus fusogenic peptide domain comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine or 10 non-contiguous amino acid deletions relative to SEQ ID NO: 244, SEQ ID NO: 245, SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 250, SEQ ID NO: 251 , SEQ ID NO: 252 or SEQ ID NO: 253.
- an avulavirus fusogenic peptide domain comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine or 10 non-contiguous amino acid additions relative to SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261 , SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265 or SEQ ID NO: 266.
- an avulavirus fusogenic peptide domain comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine or 10 contiguous amino acid deletions relative to SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 261 , SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265 or SEQ ID NO: 266.
- a metapneumovirus fusogenic peptide domain comprises amino acids a non-naturally occurring metapneumovirus fusogenic peptide domain variant of SEQ ID NO: 269, such as, e.g., a conservative metapneumovirus fusogenic peptide domain variant of SEQ ID NO: 269, a non-conservative metapneumovirus fusogenic peptide domain variant of SEQ ID NO: 269, an active metapneumovirus fusogenic peptide domain fragment of SEQ ID NO: 269, or any combination thereof.
- a conservative metapneumovirus fusogenic peptide domain variant of SEQ ID NO: 269 such as, e.g., a conservative metapneumovirus fusogenic peptide domain variant of SEQ ID NO: 269, a non-conservative metapneumovirus fusogenic peptide domain variant of SEQ ID NO: 269, an active metapneumovirus fusogenic peptide domain fragment of SEQ
- an endorphin- ⁇ variant derived from SEQ ID NO: 17 will have at least one amino acid difference, such as, e.g., an amino acid substitution, deletion or addition, as compared to SEQ ID NO: 17;
- a Dymorphin A variant derived from SEQ ID NO: 21 will have at least one amino acid difference, such as, e.g., an amino acid substitution, deletion or addition, as compared to SEQ ID NO: 21 ;
- a nociceptin variant derived from SEQ ID NO: 52 will have at least one amino acid difference, such as, e.g., an amino acid substitution, deletion or addition, as compared to SEQ ID NO: 52;
- a galanin variant derived from SEQ ID NO: 72 will have at least one amino acid difference, such as, e.g., an amino acid substitution, deletion or addition, as compared to SEQ ID NO: 72;
- an adrenomedullary peptide variant derived from SEQ ID NO: 83 will have at least one amino acid
- a naturally occurring altered targeting domain variant may substitute one or more amino acids, two or more amino acids, three or more amino acids, four or more amino acids, five or more amino acids, ten or more amino acids, 20 or more amino acids, 30 or more amino acids, 40 or more amino acids, 50 or more amino acids or 100 or more amino acids from the reference altered targeting domain on which the naturally occurring altered targeting domain variant is based.
- a peptide comprising an altered targeting domain has, e.g., at most one, two, three, four or five non-contiguous amino acid deletions relative to amino acids 1 -12, amino acids 6-22, amino acids 8-22 or amino acids 1 -22 of SEQ ID NO: 172; amino acids 1-12, amino acids 6-22, amino acids 8-22 or amino acids 1 -22 of SEQ ID NO: 173; amino acids 1 - 12, amino acids 6-22, amino acids 8-22 or amino acids 1-22 of SEQ ID NO: 174; amino acids 1 -12, amino acids 6-22, amino acids 8-22 or amino acids 1-22 of SEQ ID NO: 175; amino acids 1 -12, amino acids 6- 22, amino acids 8-22 or amino acids 1 -22 of SEQ ID NO: 176; or amino acids 1-12, amino acids 6-22, amino acids 8-22 or amino acids 1 -22 of SEQ ID NO: 177.
- a peptide comprising an altered targeting domain has, e.g., at least one, two, three, four or five contiguous amino acid substitutions relative to amino acids 1 -12, amino acids 6-22, amino acids 8-22 or amino acids 1 -22 of SEQ ID NO: 172; amino acids 1 -12, amino acids 6-22, amino acids 8-22 or amino acids 1-22 of SEQ ID NO: 173; amino acids 1 -12, amino acids 6- 22, amino acids 8-22 or amino acids 1 -22 of SEQ ID NO: 174; amino acids 1 -12, amino acids 6-22, amino acids 8-22 or amino acids 1-22 of SEQ ID NO: 175; amino acids 1-12, amino acids 6-22, amino acids 8- 22 or amino acids 1 -22 of SEQ ID NO: 176; or amino acids 1 -12, amino acids 6-22, amino acids 8-22 or amino acids 1 -22 of SEQ ID NO: 177.
- a dynorphin comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten non-contiguous amino acid substitutions relative to SEQ ID NO: 21 , SEQ ID NO: 30 or SEQ ID NO: 46.
- a dynorphin comprising an altered targeting domain has, e.g., at most one, two, three, four, five, six, seven, eight, nine or ten non-contiguous amino acid substitutions relative to SEQ ID NO: 21 , SEQ ID NO: 30 or SEQ ID NO: 46.
- a dynorphin comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid additions relative to SEQ ID NO: 21 , SEQ ID NO: 30 or SEQ ID NO: 46.
- a dynorphin comprising an altered targeting domain has, e.g., at most one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid additions relative to SEQ ID NO: 21 , SEQ ID NO: 30 or SEQ ID NO: 46.
- a nociceptin comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid substitutions relative to SEQ ID NO: 52, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61 .
- a nociceptin comprising an altered targeting domain has, e.g., at most one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid substitutions relative to SEQ ID NO: 52, SEQ ID NO: 59, SEQ ID NO: 60 or SEQ ID NO: 61 .
- an adrenocorticotropin comprising an altered targeting domain has, e.g., at least one, two, three, four or five contiguous amino acid deletions relative to SEQ ID NO: 65 or SEQ ID NO: 66. In yet other aspects of this embodiment, an adrenocorticotropin comprising an altered targeting domain has, e.g., at most one, two, three, four or five contiguous amino acid deletions relative to SEQ ID NO: 65 or SEQ ID NO: 66.
- a melanocortin peptide comprising an altered targeting domain is a lipotropin.
- a lipotropin comprising an altered targeting domain is derived from a ⁇ -lipotropin ( ⁇ -LPH) or a ⁇ -lipotropin ( ⁇ -LPH).
- a lipotropin comprising an altered targeting domain is SEQ ID NO: 67 or SEQ ID NO: 68.
- a chromogranin A peptide comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid additions relative to SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80 or SEQ ID NO: 81 .
- a chromogranin B peptide comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid substitutions relative to SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85 or SEQ ID NO: 86.
- a chromogranin B peptide comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid additions relative to SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85 or SEQ ID NO: 86.
- a chromogranin C peptide comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid substitutions relative to SEQ ID NO: 87.
- a chromogranin C peptide comprising an altered targeting domain has, e.g., at most one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid substitutions relative to SEQ ID NO: 87.
- an altered targeting domain disclosed in the present specification is, e.g., a tachykinin peptide, such as, e.g., a Substance P, a neuropeptide K (NPK), a neuropeptide gamma (NP gamma), a neurokinin A (NKA; Substance K, neurokinin alpha, neuromedin L), a neurokinin B (NKB), a hemokinin or a endokinin.
- a tachykinin peptide such as, e.g., a Substance P, a neuropeptide K (NPK), a neuropeptide gamma (NP gamma), a neurokinin A (NKA; Substance K, neurokinin alpha, neuromedin L), a neurokinin B (NKB), a hemokinin or a endokinin.
- a tachykinin peptide such as, e.g., a Substance P, a neuropeptide K
- a cholecystokinin peptide comprising an altered targeting domain has, e.g., at least 70% amino acid identity with SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ IDNO: 114 or SEQ ID NO: 115, at least 75% amino acid identity with SEQ IDNO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109
- SEQ ID NO: 106 SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 1 10, SEQ ID NO: 1 1 1 , SEQ ID NO: 1 12, SEQ ID NO: 113, SEQ ID NO: 1 14 or SEQ ID NO: 115.
- an altered targeting domain is, e.g., a Neuropeptide Y related peptide, such as, e.g., a Neuropeptide Y (NPY), a Peptide YY (PYY), Pancreatic peptide (PP) or a Pancreatic icosapeptide (PIP).
- a Neuropeptide Y related peptide such as, e.g., a Neuropeptide Y (NPY), a Peptide YY (PYY), Pancreatic peptide (PP) or a Pancreatic icosapeptide (PIP).
- NPY Neuropeptide Y
- PYYY Peptide YY
- PP Pancreatic peptide
- PIP Pancreatic icosapeptide
- a Neuropeptide Y related peptide comprising an altered targeting domain has, e.g., at most one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid deletions relative to SEQ ID NO: 116, SEQ ID NO: 1 17, SEQ ID NO: 1 18, SEQ ID NO: 119 or SEQ ID NO: 120.
- a kinin peptide comprising an altered targeting domain has, e.g., at least 70% amino acid identity with SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO: 181 , at least 75% amino acid identity with SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO: 181 , at least 80% amino acid identity with SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO: 181 , at least 85% amino acid identity with SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO: 181 , at least 90% amino acid identity with SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO: 181 or at least 95% amino acid identity with SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO
- a kinin peptide comprising an altered targeting domain has, e.g., at least one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid substitutions relative to SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO: 181 .
- a kinin peptide comprising an altered targeting domain has, e.g., at most one, two, three, four, five, six, seven, eight, nine or ten contiguous amino acid substitutions relative to SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180 or SEQ ID NO: 181 .
- a PAR peptide comprising an altered targeting domain has, e.g., at most 70% amino acid identity with amino acids 42-47, amino acids 42-55, amino acids 29-64 or amino acids 1 -64 of SEQ ID NO: 182; amino acids 35-40, amino acids 35-48, amino acids 24-59 or amino acids 1-59 of SEQ ID NO: 183; amino acids 39-44, amino acids 39-52, amino acids 26-60 or amino acids 1 -60 of SEQ ID NO: 184; amino acids 48-53, amino acids 48-61 , amino acids 35-70 or amino acids 1 -70 of SEQ ID NO: 185, at most 75% amino acid identity with amino acids 42-47, amino acids 42-55, amino acids 29-64 or amino acids 1 -64 of SEQ ID NO: 182; amino acids 35-40, amino acids 35-48, amino acids 24-59 or amino acids 1 -59 of SEQ ID NO: 183; amino acids 39-44, amino acids 39-52, amino acids 26-60 or
- a PAR peptide comprising an altered targeting domain has, e.g., at most one, two, three, four or five contiguous amino acid substitutions relative to amino acids 42-47, amino acids 42-55, amino acids 29-64 or amino acids 1 -64 of SEQ ID NO: 182; amino acids 35-40, amino acids 35-48, amino acids 24-59 or amino acids 1-59 of SEQ ID NO: 183; amino acids 39-44, amino acids 39-52, amino acids 26-60 or amino acids 1 -60 of SEQ ID NO: 184; amino acids 48-53, amino acids 48-61 , amino acids 35-70 or amino acids 1 -70 of SEQ ID NO: 185.
- a ⁇ - trefoil domain comprises four ⁇ -sheets of the first ⁇ -trefoil fold, a ⁇ -hairpin turn, four ⁇ -sheets of the second ⁇ -trefoil fold, a second ⁇ -hairpin turn four ⁇ -sheets of the third ⁇ -trefoil fold. Because the first hairpin turn is located between the fourth and fifth ⁇ -sheets of the ⁇ -trefoil domain, it is designated the ⁇ 4/ ⁇ 5 ⁇ -hairpin turn. Likewise, since the second hairpin turn is located between the eight and ninth ⁇ - sheets of the ⁇ -trefoil domain, it is designated the ⁇ 8/ ⁇ 9 ⁇ -hairpin turn.
- an altered Clostridial toxin H C c targeting region comprises a polypeptide having, e.g., at most one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 contiguous amino acid additions relative to its reference sequence.
- an altered Clostridial toxin H C c targeting region comprises a polypeptide having, e.g., at least one, two, three, four, five, six, seven, eight, nine, 10, 20, 30, 40 , 50, 100 or 200 contiguous amino acid additions relative to its reference sequence.
- a modified Clostridial toxin with an altered targeting activity comprises, in part, a protease cleavage site is located within a di-chain loop region.
- the term "di-chain loop region" means the amino acid sequence of a Clostridial toxin containing a protease cleavage site used to convert the single-chain form of a Clostridial toxin into the di-chain form.
- a modified Clostridial toxin disclosed in the present specification can comprise a flexible spacer in any and all locations with the proviso that modified Clostridial toxin is capable of performing the intoxication process.
- a flexible spacer is positioned between, e.g., an enzymatic domain and a translocation domain, an enzymatic domain and an altered targeting domain, an enzymatic domain and a protease cleavage site.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Neurology (AREA)
- Pain & Pain Management (AREA)
- Biomedical Technology (AREA)
- Neurosurgery (AREA)
- Rheumatology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Physical Education & Sports Medicine (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US80705906P | 2006-07-11 | 2006-07-11 | |
PCT/US2007/073202 WO2008008805A2 (en) | 2006-07-11 | 2007-07-11 | Modified clostridial toxins with enhanced translocation capabilities and altered targeting activity for non-clostridial toxin target cells |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2038299A2 true EP2038299A2 (de) | 2009-03-25 |
Family
ID=38924120
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP07812774A Withdrawn EP2038299A2 (de) | 2006-07-11 | 2007-07-11 | Modifizierte clostridientoxine mit verbesserten translokationsfähigkeiten und geänderter targeting-aktivität für nichtclostridientoxin-targetzellen |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP2038299A2 (de) |
JP (1) | JP2009543558A (de) |
AU (1) | AU2007272517B2 (de) |
CA (1) | CA2657521A1 (de) |
WO (1) | WO2008008805A2 (de) |
Families Citing this family (51)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7192596B2 (en) | 1996-08-23 | 2007-03-20 | The Health Protection Agency Ipsen Limited | Recombinant toxin fragments |
GB9617671D0 (en) * | 1996-08-23 | 1996-10-02 | Microbiological Res Authority | Recombinant toxin fragments |
WO2008008803A2 (en) * | 2006-07-11 | 2008-01-17 | Allergan, Inc. | Modified clostridial toxins with enhanced translocation capabilities and altered targeting activity for clostridial toxin target cells |
JP5799397B2 (ja) | 2008-06-12 | 2015-10-28 | イプセン・バイオイノベーション・リミテッドIpsen Bioinnovation Limited | 癌の抑制 |
EP3590956A1 (de) | 2008-06-12 | 2020-01-08 | Ipsen Bioinnovation Limited | Unterdrückung von neuroendokrinen erkrankungen |
GB0820970D0 (en) | 2008-11-17 | 2008-12-24 | Syntaxin Ltd | Suppression of cancer |
RU2543650C2 (ru) | 2009-03-13 | 2015-03-10 | Аллерган, Инк. | Иммунологические тесты на активность эндопептидаз с измененной нацеленностью |
US20100303789A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Chronic Neurogenic Inflammation Using Neurotrophin Retargeted Endopepidases |
US8198229B2 (en) | 2009-05-29 | 2012-06-12 | Allergan, Inc. | Methods of treating urogenital-neurological disorders using galanin retargeted endopepidases |
US20100303798A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Urogenital-Neurological Disorders Using Neurotrophin Retargeted Endopepidases |
US20100303756A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Urogenital-Neurological Disorders Using Interleukin Retargeted Endopepidases |
US20100303788A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Chronic Neurogenic Inflammation Using Galanin Retargeted Endopepidases |
US20100303791A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Chronic Neurogenic Inflammation Using Glucagon Like Hormone Retargeted Endopepidases |
US20100303783A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Urogenital-Neurological Disorders Using Tachykinin Retargeted Endopepidases |
US20100303794A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Urogenital-Neurological Disorders Using Glucagon Like Hormone Retargeted Endopepidases |
US20100303757A1 (en) * | 2009-05-29 | 2010-12-02 | Allergan, Inc. | Methods of Treating Chronic Neurogenic Inflammation Using Interleukin Retargeted Endopepidases |
EP3034511A1 (de) | 2010-01-25 | 2016-06-22 | Allergan, Inc. | Verfahren zur intrazellulären umwandlung von einkettigen proteinen in ihre zweikettige form |
MX2012013381A (es) | 2010-05-20 | 2013-02-21 | Allergan Inc | Toxinas de clostridium degradables. |
WO2012051447A1 (en) | 2010-10-14 | 2012-04-19 | Allergan, Inc. | Targeted delivery of targeted exocytosis modulators to the sphenopalatine ganglion for treatment of headache disorders |
US20120244188A1 (en) | 2011-03-25 | 2012-09-27 | Allergan, Inc. | Treatment of Sensory Disturbance Disorders |
US20120251574A1 (en) | 2011-03-28 | 2012-10-04 | Allergan, Inc. | Endopeptidase and Neurotoxin Combination Treatment of Multiple Medical Conditions |
US20120251575A1 (en) | 2011-03-28 | 2012-10-04 | Allergan, Inc. | Endopeptidase Treatment of Involuntary Movement Disorders |
US20120251573A1 (en) | 2011-03-28 | 2012-10-04 | Allergan, Inc. | Endopeptidase Treatment of Neuroendocrine Disorders |
WO2012135304A1 (en) | 2011-03-29 | 2012-10-04 | Allergan, Inc. | Vagal nerve-based disorders |
US20120251519A1 (en) | 2011-03-29 | 2012-10-04 | Allergan, Inc. | Endopeptidase Treatment of Smooth Muscle Disorders |
US20120251515A1 (en) | 2011-03-29 | 2012-10-04 | Allergan, Inc. | Endopeptidase Treatment of Cosmesis Disorders |
US20120251518A1 (en) | 2011-03-29 | 2012-10-04 | Allergan, Inc. | Endopeptidase Treatment of Sexual Dysfunction Disorders |
GB201108108D0 (en) | 2011-05-16 | 2011-06-29 | Syntaxin Ltd | Therapeutic fusion proteins |
WO2012174123A1 (en) | 2011-06-13 | 2012-12-20 | Allergan, Inc. | Treatment of psychological trauma |
US20130171122A1 (en) | 2011-12-29 | 2013-07-04 | Allergan, Inc. | Endopeptidase and neurotoxin combination treatment of bladder disorders |
US20140056870A1 (en) * | 2012-08-27 | 2014-02-27 | Allergan, Inc. | Fusion proteins |
GB201219602D0 (en) * | 2012-10-31 | 2012-12-12 | Syntaxin Ltd | Recombinant clostridium botulinum neurotoxins |
JP2015534814A (ja) * | 2012-10-31 | 2015-12-07 | イプセン バイオイノベーション リミテッド | 組換えクロストリジウムボツリヌス神経毒 |
EP2934571B1 (de) | 2012-12-18 | 2018-05-30 | Allergan, Inc. | Prophylaktische behandlung von herpesrezidiven |
EP3822286A1 (de) | 2015-01-09 | 2021-05-19 | Ipsen Bioinnovation Limited | Kationische neurotoxine |
WO2017035508A1 (en) | 2015-08-27 | 2017-03-02 | Collier R John | Compositions and methods for treatment of pain |
MX2020007596A (es) | 2018-01-29 | 2020-09-03 | Ipsen Biopharm Ltd | Neurotoxinas botulinicas para escindir una proteina de receptor de union a factor sensible a n-etilmaleimida soluble (snare) no neuronal. |
JP2021525251A (ja) | 2018-05-21 | 2021-09-24 | イプセン バイオファーム リミテッドIpsen Biopharm Limited | 骨癌誘発性アロディニアの抑制 |
GB201815817D0 (en) | 2018-09-28 | 2018-11-14 | Ispen Biopharm Ltd | Clostridial neurotoxins comprising and exogenous activation loop |
US20220016221A1 (en) | 2018-12-05 | 2022-01-20 | Ipsen Biopharm Limited | Treatment of symptoms of traumatic brain injury |
GB201900621D0 (en) | 2019-01-16 | 2019-03-06 | Ipsen Biopharm Ltd | Labelled polypeptides |
GB201914034D0 (en) | 2019-09-30 | 2019-11-13 | Ipsen Biopharm Ltd | Treatment of neurological disorders |
GB202100566D0 (en) | 2021-01-15 | 2021-03-03 | Ipsen Biopharm Ltd | Treatment of brain damage |
GB202104294D0 (en) | 2021-03-26 | 2021-05-12 | Ipsen Biopharm Ltd | Clostridial neurotoxins comprising an exogenous activation loop |
CA3211472A1 (en) | 2021-03-30 | 2022-10-06 | Mikhail KALINICHEV | Catalytically inactive clostridial neurotoxins for the treatment of pain & inflammatory disorders |
JP2024513191A (ja) | 2021-03-30 | 2024-03-22 | イプセン バイオファーム リミテッド | 疼痛及び炎症性障害の処置 |
GB202116795D0 (en) | 2021-11-22 | 2022-01-05 | Ipsen Biopharm Ltd | Treatment of visceral pain |
GB202214232D0 (en) | 2022-09-28 | 2022-11-09 | Ispen Biopharm Ltd | Clostridial neurotoxins comprising an activating exogenous protease cleavage site |
GB202214229D0 (en) | 2022-09-28 | 2022-11-09 | Ipsen Biopharm Ltd | Clostridial neurotoxins comprising an activating endosomal protease cleavage site |
WO2024069191A1 (en) | 2022-09-30 | 2024-04-04 | Ipsen Biopharm Limited | Clostridial neurotoxin for use in a treatment of bladder pain syndrome |
GB202404021D0 (en) | 2024-03-20 | 2024-05-01 | Ipsen Biopharm Ltd | Cell-based neurotoxin assay |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998007864A1 (en) | 1996-08-23 | 1998-02-26 | Microbiological Research Authority Camr (Centre For Applied Microbiology & Research) | Recombinant toxin fragments |
WO2004024909A2 (en) | 2002-09-12 | 2004-03-25 | Health Protection Agency | Recombinant colstridium neurotoxin fragments |
WO2006027207A1 (de) | 2004-09-06 | 2006-03-16 | Toxogen Gmbh | Transportprotein zum einbringen chemischer verbindungen in nervenzellen |
WO2006099590A2 (en) | 2005-03-15 | 2006-09-21 | Allergan, Inc. | Modified clostridial toxins with altered targeting capabilities for clostridial toxin target cells |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5135736A (en) * | 1988-08-15 | 1992-08-04 | Neorx Corporation | Covalently-linked complexes and methods for enhanced cytotoxicity and imaging |
WO2001014570A1 (en) * | 1999-08-25 | 2001-03-01 | Allergan Sales, Inc. | Activatable recombinant neurotoxins |
US20060024331A1 (en) * | 2004-08-02 | 2006-02-02 | Ester Fernandez-Salas | Toxin compounds with enhanced membrane translocation characteristics |
EP1784420B1 (de) * | 2004-09-01 | 2008-12-03 | Allergan, Inc. | Abbaubare clostridientoxine |
WO2008008803A2 (en) * | 2006-07-11 | 2008-01-17 | Allergan, Inc. | Modified clostridial toxins with enhanced translocation capabilities and altered targeting activity for clostridial toxin target cells |
-
2007
- 2007-07-11 EP EP07812774A patent/EP2038299A2/de not_active Withdrawn
- 2007-07-11 JP JP2009519645A patent/JP2009543558A/ja active Pending
- 2007-07-11 CA CA002657521A patent/CA2657521A1/en not_active Abandoned
- 2007-07-11 AU AU2007272517A patent/AU2007272517B2/en not_active Ceased
- 2007-07-11 WO PCT/US2007/073202 patent/WO2008008805A2/en active Application Filing
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998007864A1 (en) | 1996-08-23 | 1998-02-26 | Microbiological Research Authority Camr (Centre For Applied Microbiology & Research) | Recombinant toxin fragments |
US8017134B2 (en) | 1996-08-23 | 2011-09-13 | Syntaxin Limited | Recombinant toxin fragments |
WO2004024909A2 (en) | 2002-09-12 | 2004-03-25 | Health Protection Agency | Recombinant colstridium neurotoxin fragments |
WO2006027207A1 (de) | 2004-09-06 | 2006-03-16 | Toxogen Gmbh | Transportprotein zum einbringen chemischer verbindungen in nervenzellen |
WO2006099590A2 (en) | 2005-03-15 | 2006-09-21 | Allergan, Inc. | Modified clostridial toxins with altered targeting capabilities for clostridial toxin target cells |
Non-Patent Citations (4)
Title |
---|
HERREROS J ET AL: "C-TERMINAL HALF OF TETANUS TOXIN FRAGMENT C IS SUFFICIENT FOR NEURONAL BINDING AND INTERACTION WITH A PUTATIVE PROTEIN RECEPTOR", BIOCHEMICAL JOURNAL, THE BIOCHEMICAL SOCIETY, LONDON, GB, vol. 347, no. PART 01, 1 April 2001 (2001-04-01), pages 199 - 204, XP001010346, ISSN: 0264-6021, DOI: 10.1042/0264-6021:3470199 * |
LUDWIG C U ET AL: "Fibroblasts stimulate acinar cell proliferation through IGF-I during regeneration from acute pancreatitis.", THE AMERICAN JOURNAL OF PHYSIOLOGY JAN 1999, vol. 276, no. 1 Pt 1, January 1999 (1999-01-01), pages G193 - G198, XP003030337, ISSN: 0002-9513 * |
LUDWIG C.U. ET AL: "Fibroblasts stimulate acinar cell prolifeeration through IGF-I during regeneration from acute pancreatitis", AMERICAN JOURNAL OF PHYSIOLOGY: GASTROINTESTINAL AND LIVER PHYSIOLOGY, vol. 276, 1999, pages G193 - G198, XP003030337 |
See also references of WO2008008805A2 * |
Also Published As
Publication number | Publication date |
---|---|
WO2008008805A2 (en) | 2008-01-17 |
CA2657521A1 (en) | 2008-01-17 |
AU2007272517B2 (en) | 2013-09-26 |
WO2008008805A3 (en) | 2008-04-10 |
AU2007272517A1 (en) | 2008-01-17 |
JP2009543558A (ja) | 2009-12-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2007272517B2 (en) | Modified clostridial toxins with enhanced translocation capabilities and altered targeting activity for non-clostridial toxin target cells | |
US7993656B2 (en) | Modified clostridial toxins with enhanced translocation capabilities and altered targeting activity for clostridial toxin target cells | |
US8518417B1 (en) | Modified clostridial toxins with enhanced translocation capability and enhanced targeting activity | |
US8460682B2 (en) | Modified clostridial toxins with altered targeting capabilities for clostridial toxin target cells | |
EP2001902B1 (de) | Modifizierte clostridiale toxine mit geänderten targeting-fähigkeiten für die target-zellen von clostridialem toxin | |
US8021859B2 (en) | Modified clostridial toxins with altered targeting capabilities for clostridial toxin target cells | |
US8486422B2 (en) | Methods of activating clostridial toxins | |
US20140127784A1 (en) | Modified clostridial toxins comprising an integrated protease cleavage site-binding domain | |
US20130288336A1 (en) | Methods of activating clostridial toxins |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20090112 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR |
|
AX | Request for extension of the european patent |
Extension state: AL BA HR MK RS |
|
DAX | Request for extension of the european patent (deleted) | ||
17Q | First examination report despatched |
Effective date: 20121122 |
|
TPAC | Observations filed by third parties |
Free format text: ORIGINAL CODE: EPIDOSNTIPA |
|
TPAC | Observations filed by third parties |
Free format text: ORIGINAL CODE: EPIDOSNTIPA |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
INTG | Intention to grant announced |
Effective date: 20150226 |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
INTG | Intention to grant announced |
Effective date: 20150707 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20151118 |